Patent Application
20250230165
The invention relates to heterocyclic derivatives and their use in the treatment and prophylaxis of cancer, and to compositions containing said derivatives and processes for their preparation.
Robust repair of DNA double-strand breaks (DSBs) is essential for the maintenance of genome stability and cell viability. DSBs can be repaired by one of three main pathways: homologous recombination (HR), non-homologous end-joining (NHEJ) and alternative NHEJ (alt-NHEJ). Microhomology-mediated end-joining (MMEJ) is the most well characterised alt-NHEJ mechanism. HR-mediated repair is a high-fidelity mechanism essential for accurate error-free repair, preventing cancer-predisposing genomic stability. Conversely, NHEJ and MMEJ are error-prone pathways that can leave mutational scars at the site of repair. MMEJ can function parallel to both HR and NHEJ pathways (Truong et al. PNAS 2013, 110 (19), 7720-7725).
The survival of cancer cells, unlike normal cells, is often dependent on the mis-regulation of DNA damage response (DDR) pathways. For example, an increased dependency on one pathway (often mutagenic) to cope with either the inactivation of another one, or the enhanced replication stress resulting from increased proliferation. An aberrant DDR can also sensitise cancer cells to specific types of DNA damage, thus, defective DDR can be exploited to develop targeted cancer therapies. Crucially, cancer cells with impairment or inactivation of HR and NHEJ become hyper-dependent on MMEJ-mediated DNA repair. Genetic, cell biological and biochemical data have identified Polθ (UniProtKB-O75417 (DPOLQ_HUMAN) as the key protein in MMEJ (Kent et al. Nature Structural & Molecular Biology (2015), 22(3), 230-237, Mateos-Gomez et al. Nature (2015), 518(7538), 254-257). Polθ is multifunctional enzyme, which comprises an N-terminal helicase domain (SF2 HEL308-type) and a C-terminal low-fidelity DNA polymerase domain (A-type) (Wood & Doublié DNA Repair (2016), 44, 22-32). Both domains have been shown to have concerted mechanistic functions in MMEJ. The helicase domain mediates the removal of RPA protein from ssDNA ends and stimulates annealing. The polymerase domain extends the ssDNA ends and fills the remaining gaps.
Therapeutic inactivation of Polθ would thus disable the ability of cells to perform MMEJ and provide a novel targeted strategy in an array of defined tumour contexts. Firstly, Polθ has been shown to be essential for the survival of HR-defective (HRD) cells (e.g. synthetic lethal with FA/BRCA-deficiency) and is up-regulated in HRD tumour cell lines (Ceccaldi et al. Nature (2015), 518(7538), 258-262). In vivo studies also show that Polθ is significantly over-expressed in subsets of HRD ovarian, uterine and breast cancers with associated poor prognosis (Higgins et al. Oncotarget (2010), 1, 175-184, Lemée et al. PNAS (2010), 107(30), 13390-13395, Ceccaldi et al. (2015), supra). Importantly, Polθ is largely repressed in normal tissues but has been shown to be upregulated in matched cancer samples thus correlating elevated expression with disease (Kawamura et al. International Journal of Cancer (2004), 109(1), 9-16). Secondly, its suppression or inhibition confers radio-sensitivity in tumour cells. Finally, Polθ inhibition could conceivably prevent the MMEJ-dependent functional reversion of BRCA2 mutations that underlies the emergence of cisplatin and PARPi resistance in tumours.
There is therefore a need to provide effective Polθ inhibitors for the treatment of cancer.
According to a first aspect of the invention, there is provided a compound of formula (I):
The term ‘halo’ or ‘halogen’ as used herein refers to fluorine, chlorine, bromine or iodine.
The term ‘cyano’ as used herein refers to a group where a carbon atom is triple bonded to a nitrogen atom.
The term ‘C1-6 alkyl’ as used herein as a group or part of a group refers to a linear or branched saturated hydrocarbon group containing from 1 to 6 carbon atoms. Examples of such groups include methyl, ethyl, n-propyl, iso-propyl, butyl, iso-butyl, tert-butyl, pentyl, hexyl and the like.
The term ‘C2-6 alkenyl’ as used herein as a group or part of a group refers to a linear or branched unsaturated hydrocarbon group containing from 2 to 6 carbon atoms and at least one double bond. Examples of such groups include ethenyl, propenyl, butenyl, pentenyl, hexenyl and the like.
The term ‘C2-6alkynyl’ as used herein as a group or part of a group refers to a linear or branched hydrocarbon group having from 2 to 6 carbon atoms, respectively, and containing a carbon carbon triple bond. Examples of such groups include C3-4alkynyl or C3-6alkynyl groups such as ethynyl and 2 propynyl (propargyl) groups.
The term ‘C1-6 alkoxy’ as used herein as a group or part of a group refers to a C1-6 alkyl group which contains an oxygen atom wherein C1-6 alkyl is as defined herein. Examples of such groups include methoxy, ethoxy or propoxy.
The term “C1-Cx alkylene” as used herein as a group or part of a group refers to a —(CH2)1-x group. Examples of such groups include methylene, ethylene, propylene and butylene.
The term ‘haloC1-6alkyl’ as used herein as a group or part of a group refers to a C1-6 alkyl group as defined herein wherein one or more than one hydrogen atom is replaced with a halogen. The term ‘haloC1-6alkyl’ therefore includes monohaloC1-6 alkyl and also polyhaloC1-6 alkyl. There may be one, two, three or more hydrogen atoms replaced with a halogen, so the haloC1-6alkyl may have one, two, three or more halogens. Examples of such groups include fluoroethyl, fluoromethyl, trifluoromethyl or trifluoroethyl and the like.
The term “C3-8 cycloalkyl” as used herein refers to a saturated monocyclic hydrocarbon ring of 3 to 8 carbon atoms. Examples of such groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl and the like.
The term ‘oxo’ as used herein refers to the group ═O.
The term ‘hydroxy’ as used herein refers to the group —OH. The term ‘carbocyclyl’ includes aromatic and non-aromatic ring systems comprising solely carbon and hydrogen (i.e. not containing any heteroatoms). Carbocyclyl ring systems have from 5 to 12 ring members, more usually from 5 to 10 ring members. In one embodiment, carbocyclyl includes an aryl ring which refers to carbocyclyl aromatic groups including phenyl, naphthyl, indanyl, indenyl, and tetrahydronaphthyl groups. The term “aryl” embraces polycyclic (e.g. bicyclic) ring systems wherein one or more rings are non-aromatic, provided that at least one ring is aromatic. Examples of polycyclic (e.g. bicyclic) aryl groups containing an aromatic ring and a non-aromatic ring include indanyl groups. In such polycyclic systems, the group may be attached by the aromatic ring, or by a non-aromatic ring. One particular example of an aromatic carbocyclic ring system includes phenyl.
In an alternative embodiment, carbocyclyl includes a non-aromatic carbocyclic group such as cycloalkyl and cycloalkenyl groups as defined herein and spiro and bridged derivatives thereof. Particular examples of non-aromatic carbocyclic ring system include cyclobutyl, cyclohexyl, 2,3-dihydro-1H-inden-2-yl and bicyclo[1.1.1]pentan-1-yl.
The term ‘heteroaryl’ as used herein refers to a monocyclic or bicyclic aromatic ring system containing for example 3 to 12 ring members. Each ring may contain up to five heteroatoms typically selected from nitrogen, sulfur and oxygen. Particular examples of heteroaryl include imidazolyl, pyrazolyl, triazolyl, pyridinyl, pyrimidinyl, oxazolyl and oxadiazolyl.
The term ‘heterocyclyl’ as used herein refers to a monocyclic, bicyclic or tricyclic non-aromatic, partially saturated or fully saturated ring system containing for example 3 to 12 ring members. Each ring may contain up to five heteroatoms typically selected from nitrogen, sulfur and oxygen. Particular examples of bicyclic or tricyclic heterocyclyl include monocyclic or bicyclic heterocyclic rings which may be fused to monocyclic carbocyclic and heteroaromatic rings.
Particular examples of ‘heterocyclyl’ include morpholine, piperidine (e.g. piperidin-1-yl, piperidin-2-yl, piperidin-3-yl and piperidin-4-yl), piperidinone, pyrrolidine (e.g. pyrrolidin-1-yl, pyrrolidin-2-yl and pyrrolidin-3-yl), pyrrolidone, azetidine, pyran (2H-pyran or 4H-pyran), dihydrothiophene, dihydropyran, dihydrofuran, dihydrothiazole, tetrahydrofuran, tetrahydrothiophene, dioxane, diazepane, tetrahydropyran (e.g. tetrahydropyran-4-yl), tetrahydropyridine, imidazoline, imidazolidinone, oxazoline, thiazoline, pyrazolin-2-yl, pyrazolidine, piperazinone, piperazine, hexahydropyrrolo[3,4-b][1,4]oxazin-6(2H)-yl, octahydro-2H-pyrazino[1,2-a]pyrazin-2-yl, octahydro-2H-pyrazino[1,2-a]pyrazin-3-yl, octahydro-5H-pyrrolo[3,4-c]pyridin-5-yl, tetrahydro-1H-imidazo[4,5-c]pyridin-1-yl, tetrahydro-3H-imidazo[4,5-c]pyridin-3-yl, isoindolin-1-yl, isoindolin-5-yl, tetrahydroisoquinolin-6-yl, tetrahydro-[1,2,4]triazolo[1,5-a]pyrazin-2-yl, tetrahydro-[1,2,4]triazolo[4,3-a]pyrazin-3-yl, tetrahydroimidazo[1,2-a]pyrazin-2-yl, dihydro-5H-pyrrolo[3,4-b]pyrazin-2-yl, dihydro-5H-pyrrolo[3,4-b]pyridin-3-yl, dihydro-5H-pyrrolo[3,4-b]pyridin-2-yl, dihydro-5H-pyrrolo[3,4-b]pyridin-7-yl, dihydro-5H-pyrrolo[3,4-b]pyridin-5-yl, tetrahydropyrrolo[3,4-d]imidazol-2-yl, dihydro-2H-benzo[b][1,4]oxazin-6-yl, dihydro-2H-benzo[b][1,4]oxazin-7-yl, dihydro-4H-pyrrolo[3,4-d]imidazol-1-yl, dihydro-1H-pyrrolo[3,4-c]pyridin-6-yl, dihydro-5H-pyrrolo[3,4-d]pyrimidin-2-yl, dihydro-4H-pyrrolo[3,4-d]thiazol-2-yl, dihydro-4H-pyrrolo[3,4-d]oxazo-2-yl, tetrahydropyrrolo[3,4-c]pyrazol-3-yl, tetrahydroimidazo[1,2-a]pyridin-6-yl, tetrahydroimidazo[1,2-a]pyridin-7-yl, and tetrahydropyrazolo[1,5-a]pyrazin-2-yl.
It will be appreciated that the term ‘heterocyclyl” includes reference to spiro and bridged heterocyclic derivatives. Examples of such spiro and bridged heterocyclic derivatives include: hexahydropyrrolo[2,3-c]pyrrolidinyl, diazaspiro[3.3]heptanyl, azaspiro[3.3]heptan-6-yl, diazaspiro[3.4]octanyl, diazaspiro[4.4]nonyl, oxa-azaspiro[3.4]octanyl, oxa-azaspiro[4.4]nonyl, tetrahydrofuro[3,4-c]pyrrolidinyl, oxa-azaspiro[3.3]heptyl, diazaspiro[3.5]nonanyl, diazaspiro[4.4]nonanyl, diazaspiro[4.5]decanyl, diazaspiro[3.5]nonanyl, octahydro-naphthyridinyl, tetrahydropyrazino-oxazinyl, oxadiazaspiro[3.4]octanyl, oxadiazaspiro[3.5]nonanyl, oxadiazaspiro[4.5]decanyl, oxadiazospiro[5.5]undecanyl, triazaspiro[3.5]nonanyl, diazabicyclo[2.2.1]heptan-2-yl, diazabicyclo[3.1.1]heptan-6-yl, diazabicyclo[3.2.1]octan-8-yl, oxabicyclo[2.2.1]heptanyl, oxa-2,8-diazaspiro[3.5]nonan-8-yl, oxa-3,7-diazabicyclo[3.3.1]nonan-3-yl, tetrahydro-1′H-spiro[azetidine-3,4′-pyrrolo[1,2-a]pyrazin]-2′(3′H)-yl, tetrahydro-1′H-spiro[azetidine-3,6′-pyrazino[2,1-c][1,4]oxazin]-8′(7′H)-yl, triazaspiro[3.5]nonan-8-yl, spiro[azetidine-3,3′-imidazo[1,2-a]imidazol]-1′(2′H)-yl and spiro[azetidine-3,5′-imidazo[1,2-a]pyrazin]-7′(8′H)-yl). The term ‘optionally substituted’ as used herein refers to a group which may be substituted or unsubstituted by a substituent as herein defined.
According to one particular aspect of the invention which may be mentioned, there is provided a compound of formula (I):
In one embodiment, n represents 0, 1, 2 or 3.
In one embodiment, R1 represents C1-6 alkyl (such as methyl), halogen (such as fluorine or chlorine) or C3-8 cycloalkyl (such as cyclopropyl). In a further embodiment, R1 represents C1-6 alkyl (such as methyl) or halogen (such as fluorine or chlorine). In a yet further embodiment, R1 represents halogen (such as fluorine or chlorine). In an alternative embodiment, R1 represents C1-6 alkyl (such as methyl). In a further embodiment, R1 represents methyl.
In a further embodiment, n represents 0.
In a further embodiment, n represents 1. In a yet further embodiment, n represents 1 and R1 represents C1-6 alkyl (such as methyl) or halogen (such as fluorine or chlorine). In a yet further embodiment, n represents 1 and R1 represents methyl, fluorine or chlorine. In a yet further embodiment, n represents 1 and R1 represents C1-6 alkyl (such as methyl). In a still yet further embodiment, n represents 1 and R1 represents methyl.
In an alternative embodiment, n represents 2. In a yet further embodiment, n represents 2 and R1 represents C1-6 alkyl (such as methyl), halogen (such as fluorine or chlorine) or C3-8 cycloalkyl (such as cyclopropyl). In a yet further embodiment, n represents 2 and R1 represents methyl, fluorine, chlorine, or cyclopropyl. In a still yet further embodiment, n represents 2 and:
In an alternative embodiment, n represents 3. In a yet further embodiment, n represents 3 and R1 represents halogen (such as fluorine or chlorine). In a yet further embodiment, n represents 3 and R1 represents fluorine or chlorine. In a still yet further embodiment, n represents 3 and two R1 groups represent fluorine and one R1 group represents chlorine.
In one embodiment, R2 represents -RingA.
In one embodiment, RingA represents carbocyclyl (such as phenyl, cyclobutyl, 2,3-dihydro-1H-inden-2-yl or bicyclo[1.1.1]pentan-1-yl), a heterocyclyl ring (such as azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, isoindolin-1-yl, isoindolin-5-yl, tetrahydroisoquinolin-6-yl, diazaspiro[3.4]octan-6-yl, octahydro-2H-pyrazino[1,2-a]pyrazin-2-yl, octahydro-5H-pyrrolo[3,4-c]pyridin-5-yl, 5-oxa-2,8-diazaspiro[3.5]nonan-8-yl, 2,6-diazaspiro[3.4]octane, 9-oxa-3,7-diazabicyclo[3.3.1]nonan-3-yl, tetrahydro-1H-imidazo[4,5-c]pyridin-1-yl, tetrahydro-1′H-spiro[azetidine-3,4′-pyrrolo[1,2-a]pyrazin]-2′(3′H)-yl, tetrahydro-1′H-spiro[azetidine-3,6′-pyrazino[2,1-c][1,4]oxazin]-8′(7′H)-yl, tetrahydro-3H-imidazo[4,5-c]pyridin-3-yl, tetrahydro-[1,2,4]triazolo[1,5-a]pyrazin-2-yl, tetrahydroimidazo[1,2-a]pyrazin-2-yl, tetrahydropyrazolo[1,5-a]pyrazin-2-yl, octahydro-2H-pyrazino[1,2-a]pyrazin-3-yl, tetrahydro-[1,2,4]triazolo[4,3-a]pyrazin-3-yl, 2,5,8-triazaspiro[3.5]nonan-8-yl, spiro[azetidine-3,3′-imidazo[1,2-a]imidazol]-1′(2′H)-yl, dihydro-5H-pyrrolo[3,4-b]pyrazin-2-yl, dihydro-5H-pyrrolo[3,4-b]pyridin-3-yl, dihydro-5H-pyrrolo[3,4-b]pyridin-2-yl, dihydro-5H-pyrrolo[3,4-b]pyridin-7-yl, dihydro-5H-pyrrolo[3,4-b]pyridin-5-yl, tetrahydropyrrolo[3,4-d]imidazol-2-yl, dihydro-2H-benzo[b][1,4]oxazin-6-yl, dihydro-2H-benzo[b][1,4]oxazin-7-yl, dihydro-4H-pyrrolo[3,4-d]imidazol-1-yl, diazabicyclo[3.1.1]heptan-6-yl, diazabicyclo[3.2.1]octan-8-yl, diazabicyclo[2.2.1]heptan-2-yl, dihydro-1H-pyrrolo[3,4-c]pyridin-6-yl, dihydro-5H-pyrrolo[3,4-d]pyrimidin-2-yl, dihydro-4H-pyrrolo[3,4-d]thiazol-2-yl, dihydro-4H-pyrrolo[3,4-d]oxazo-2-yl, tetrahydropyrrolo[3,4-c]pyrazol-3-yl, diazaspiro[3.3]heptan-6-yl, tetrahydroimidazo[1,2-a]pyridin-6-yl, tetrahydroimidazo[1,2-a]pyridin-7-yl, or spiro[azetidine-3,5′-imidazo[1,2-a]pyrazin]-7′(8′H)-yl) or a heteroaryl ring (such as pyridyl or pyrimidinyl), each being substituted by a
In a further embodiment, RingA represents carbocyclyl (such as phenyl or bicyclo[1.1.1]pentan-1-yl), a heterocyclyl ring (such as azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, isoindolin-5-yl, tetrahydroisoquinolin-6-yl, diazaspiro[3.4]octan-6-yl, octahydro-2H-pyrazino[1,2-a]pyrazin-2-yl, octahydro-5H-pyrrolo[3,4-c]pyridin-5-yl, 5-oxa-2,8-diazaspiro[3.5]nonan-8-yl, 2,6-diazaspiro[3.4]octane, 9-oxa-3,7-diazabicyclo[3.3.1]nonan-3-yl, tetrahydro-1H-imidazo[4,5-c]pyridin-1-yl, tetrahydro-1′H-spiro[azetidine-3,4′-pyrrolo[1,2-a]pyrazin]-2′(3′H)-yl, tetrahydro-1′H-spiro[azetidine-3,6′-pyrazino[2,1-c][1,4]oxazin]-8′(7′H)-yl, tetrahydro-3H-imidazo[4,5-c]pyridin-3-yl, tetrahydro-[1,2,4]triazolo[1,5-a]pyrazin-2-yl, tetrahydroimidazo[1,2-a]pyrazin-2-yl, tetrahydropyrazolo[1,5-a]pyrazin-2-yl, octahydro-2H-pyrazino[1,2-a]pyrazin-3-yl, tetrahydro-[1,2,4]triazolo[4,3-a]pyrazin-3-yl, 2,5,8-triazaspiro[3.5]nonan-8-yl, spiro[azetidine-3,3′-imidazo[1,2-a]imidazol]-1′(2′H)-yl or spiro[azetidine-3,5′-imidazo[1,2-a]pyrazin]-7′(8′H)-yl) or a heteroaryl ring (such as pyridyl), each being substituted by a
In a further embodiment, RingA represents a heterocyclyl ring (such as azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, isoindolin-1-yl, isoindolin-5-yl, tetrahydroisoquinolin-6-yl, diazaspiro[3.4]octan-6-yl, octahydro-2H-pyrazino[1,2-a]pyrazin-2-yl, octahydro-5H-pyrrolo[3,4-c]pyridin-5-yl, 5-oxa-2,8-diazaspiro[3.5]nonan-8-yl, 2,6-diazaspiro[3.4]octane, 9-oxa-3,7-diazabicyclo[3.3.1]nonan-3-yl, tetrahydro-1H-imidazo[4,5-c]pyridin-1-yl, tetrahydro-1′H-spiro[azetidine-3,4′-pyrrolo[1,2-a]pyrazin]-2′(3′H)-yl, tetrahydro-1′H-spiro[azetidine-3,6′-pyrazino[2,1-c][1,4]oxazin]-8′(7′H)-yl, tetrahydro-3H-imidazo[4,5-c]pyridin-3-yl, tetrahydro-[1,2,4]triazolo[1,5-a]pyrazin-2-yl, tetrahydroimidazo[1,2-a]pyrazin-2-yl, tetrahydropyrazolo[1,5-a]pyrazin-2-yl, octahydro-2H-pyrazino[1,2-a]pyrazin-3-yl, tetrahydro-[1,2,4]triazolo[4,3-a]pyrazin-3-yl, 2,5,8-triazaspiro[3.5]nonan-8-yl, spiro[azetidine-3,3′-imidazo[1,2-a]imidazol]-1′(2′H)-yl, dihydro-5H-pyrrolo[3,4-b]pyrazin-2-yl, dihydro-5H-pyrrolo[3,4-b]pyridin-3-yl, dihydro-5H-pyrrolo[3,4-b]pyridin-2-yl, dihydro-5H-pyrrolo[3,4-b]pyridin-7-yl, dihydro-5H-pyrrolo[3,4-b]pyridin-5-yl, tetrahydropyrrolo[3,4-d]imidazol-2-yl, dihydro-2H-benzo[b][1,4]oxazin-6-yl, dihydro-2H-benzo[b][1,4]oxazin-7-yl, dihydro-4H-pyrrolo[3,4-d]imidazol-1-yl, diazabicyclo[3.1.1]heptan-6-yl, diazabicyclo[3.2.1]octan-8-yl, diazabicyclo[2.2.1]heptan-2-yl, dihydro-1H-pyrrolo[3,4-c]pyridin-6-yl, dihydro-5H-pyrrolo[3,4-d]pyrimidin-2-yl, dihydro-4H-pyrrolo[3,4-d]thiazol-2-yl, dihydro-4H-pyrrolo[3,4-d]oxazo-2-yl, tetrahydropyrrolo[3,4-c]pyrazol-3-yl, diazaspiro[3.3]heptan-6-yl, tetrahydroimidazo[1,2-a]pyridin-6-yl, tetrahydroimidazo[1,2-a]pyridin-7-yl, or spiro[azetidine-3,5′-imidazo[1,2-a]pyrazin]-7′(8′H)-yl) each being substituted by a
In a further embodiment, RingA represents dihydro-5H-pyrrolo[3,4-b]pyridinyl, such as dihydro-5H-pyrrolo[3,4-b]pyridin-3-yl, dihydro-5H-pyrrolo[3,4-b]pyridin-2-yl, dihydro-5H-pyrrolo[3,4-b]pyridin-7-yl, or dihydro-5H-pyrrolo[3,4-b]pyridin-5-yl each being substituted by a
In a yet further embodiment, RingA represents dihydro-5H-pyrrolo[3,4-b]pyridinyl, such as dihydro-5H-pyrrolo[3,4-b]pyridin-3-yl, dihydro-5H-pyrrolo[3,4-b]pyridin-2-yl, dihydro-5H-pyrrolo[3,4-b]pyridin-7-yl, or dihydro-5H-pyrrolo[3,4-b]pyridin-5-yl each being substituted by a —COCH═CH2.
In a still yet further embodiment, RingA represents dihydro-5H-pyrrolo[3,4-b]pyridin-2-yl substituted by a —COCH═CH2.
In a still yet further embodiment, RingA represents 6,7-dihydro-5H-pyrrolo[3,4-b]pyridin-2-yl substituted by a —COCH═CH2.
In an alternative embodiment, R2 represents -RingB-Y-RingC.
In one embodiment, RingB represents carbocyclyl (such as cyclohexyl or phenyl), heterocyclyl (such as azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, diazepanyl or hexahydropyrrolo[3,4-b][1,4]oxazin-6(2H)-yl) or heteroaryl (such as imidazolyl, pyrazolyl, triazolyl, pyridinyl, pyrimidinyl, oxazolyl or oxadiazolyl), each of which may be optionally substituted by one or more (e.g. 1, 2 or 3) substituents selected from hydroxy, oxo, C1-6 alkyl (such as methyl), haloC1-6 alkyl (such as —CH2—F), cyano or halogen (such as fluorine).
In a further embodiment, RingB represents cyclohexyl, phenyl, azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, diazepanyl, hexahydropyrrolo[3,4-b][1,4]oxazin-6(2H)-yl, imidazolyl, pyrazolyl, triazolyl, pyridinyl, pyrimidinyl, oxazolyl or oxadiazolyl, each of which may be optionally substituted by one or more (e.g. 1, 2 or 3) substituents selected from hydroxy, oxo, methyl, —CH2—F, cyano or fluorine.
In a further embodiment, RingB represents carbocyclyl (such as cyclohexyl or phenyl), heterocyclyl (such as azetidinyl, pyrrolidinyl, piperazinyl, diazepanyl or hexahydropyrrolo[3,4-b][1,4]oxazin-6(2H)-yl) or heteroaryl (such as imidazolyl, pyrazolyl, triazolyl, pyridinyl, pyrimidinyl, oxazolyl or oxadiazolyl), each of which may be optionally substituted by one or more (e.g. 1, 2 or 3) substituents selected from hydroxy, oxo, C1-6 alkyl (such as methyl), haloC1-6 alkyl (such as —CH2—F), cyano or halogen (such as fluorine).
In one embodiment, Y represents a bond, —O—, —NHCO—, CO, or a C1-C6 alkylene group optionally substituted by one or more C1-6 alkyl, C2-6 alkenyl, —NRx, O, hydroxy or CO groups (such as —CO—C(═CH2)—CH2— or —NHCO—CH═CH—CH2—). In a further embodiment, Y represents a bond, —O—, —NHCO—, CO, or a C1-C6 alkylene group optionally substituted by one or more C1-6 alkyl, C2-6 alkenyl, —NRx, O, hydroxy or CO groups (such as —CO—C(═CH2)—CH2—).
In a further embodiment, Y represents a bond. In an alternative embodiment, Y represents —O—.
In one embodiment, RingC represents a monocyclic heterocyclyl ring having at least one nitrogen atom. In a further embodiment, RingC represents azetidinyl, pyrrolidinyl, piperazinyl, piperidinyl, morpholinyl or tetrahydropyridinyl, each of which being substituted by a halogen (such as fluorine), C1-6 alkyl (such as methyl), —CO—C1-6 alkyl (such as —CO-methyl) and/or a —(CH2)m—CO—C2-6 alkenyl optionally substituted by an —NRxRy group (such as —COCH═CH2, —CO—C(═CH2)—CH2—N(Me)2, —CO—C(═CH-Me)-CH2—N(Me)2, or —CO—CH═CH—CH2—N(Me)2).
In a further embodiment, RingC represents azetidinyl, pyrrolidinyl, piperazinyl, piperidinyl, morpholinyl or tetrahydropyridinyl, each of which being substituted by a fluorine, methyl, —CO-methyl, and/or a —COCH═CH2, —CO—C(═CH2)—CH2—N(Me)2, —CO—C(═CH-Me)-CH2—N(Me)2, or —CO—CH═CH—CH2—N(Me)2.
In a further embodiment, RingC represents azetidinyl, pyrrolidinyl, piperazinyl, piperidinyl, morpholinyl or tetrahydropyridinyl, each of which being substituted by:
In a further embodiment, RingC represents azetidinyl, pyrrolidinyl, piperazinyl or tetrahydropyridinyl, each of which being substituted by a C1-6 alkyl (such as methyl) and/or a —(CH2)m—CO—C2-6 alkenyl optionally substituted by an —NRxRy group (such as —COCH═CH2, —CO—C(═CH2)—CH2—N(Me)2, —CO—C(═CH-Me)-CH2—N(Me)2, or —CO—CH═CH—CH2—N(Me)2).
In one embodiment, X represents a bond or a C1-C4 alkylene group optionally substituted by one or more C1-6 alkyl (such as methyl), —NRx (such as —NH or —N-methyl), O, hydroxy, halogen (such as fluorine) or CO groups.
In a further embodiment, X represents a bond or a C1-C4 alkylene group optionally substituted by one or more C1-6 alkyl (such as methyl), —NRx (such as —NH or —N-methyl), O, hydroxy or CO groups.
In a further embodiment, X represents a bond.
In an alternative embodiment, X represents a C1-C4 alkylene group (such as —CH2—, —(CH2)2—, —(CH2)3—, —(CH2)4—, or —(CH2)5—) optionally substituted by one or more C1-6 alkyl (such as —(CH2)—C(Me)-), O (such as —(CH2)2—O—, —(CH2)3—O— or —(CH2)2O—CH2—) or CO (such as —(CH2)—CO— or —(CH2)2—CO—) —NRx (such as —(CH2)2—NH—, —(CH2)3—NH—, —(CH2)2—N(Me)- or —(CH2)3—N(Me)-) groups or a CO and —NRx group (such as —(CH2)2—N(Me)-CO—) or a hydroxy and —NRx group (such as —(CH2)—CH(OH)—CH2—N(Me)- or halogen (such as —(CH2)3—CHF—, —(CH2)2—CHF—CH2—, or —CH2—CHF—(CH2)2—).
In a further embodiment, X represents a C1-C4 alkylene group (such as —CH2—, —(CH2)2—, —(CH2)3— or —(CH2)4—) optionally substituted by one or more C1-6 alkyl (such as —(CH2)—C(Me)-), O (such as —(CH2)2—O—, —(CH2)3—O— or —(CH2)2O—CH2—) or CO (such as —(CH2)—CO— or —(CH2)2—CO—) —NRx (such as —(CH2)2—NH—, —(CH2)3—NH—, —(CH2)2—N(Me)- or —(CH2)3—N(Me)-) groups or a CO and —NRx group (such as —(CH2)2—N(Me)-CO—) or a hydroxy and —NRx group (such as —(CH2)—CH(OH)—CH2—N(Me)-).
In a yet further embodiment, X represents a bond, —CH2—, —(CH2)2—, —(CH2)3—, —(CH2)4—, —(CH2)—C(Me)-, —(CH2)2—O—, —(CH2)3—O—, —(CH2)2—O—CH2—, —(CH2)—CO—, —(CH2)2—CO—, —(CH2)2—NH—, —(CH2)3—NH—, —(CH2)2—N(Me)-, —(CH2)3—N(Me)-, —(CH2)2—N(Me)-CO—, —(CH2)—CH(OH)—CH2—N(Me)-, —(CH2)3—CHF—, —(CH2)2—CHF—CH2—, or —CH2—CHF—(CH2)2—.
In a yet further embodiment, X represents a bond, —CH2—, —(CH2)2—, —(CH2)3—, —(CH2)4—, —(CH2)—C(Me)-, —(CH2)2—O—, —(CH2)3—O—, —(CH2)2—O—CH2—, —(CH2)—CO—, —(CH2)2—CO—, —(CH2)2—NH—, —(CH2)3—NH—, —(CH2)2—N(Me)-, —(CH2)3—N(Me)-, —(CH2)2—N(Me)-CO— or —(CH2)—CH(OH)—CH2—N(Me)-.
In a still yet further embodiment, X represents —CH2—.
In one embodiment, —X—R2 represents a —C1-C10 alkylene-NRxRy group, wherein said alkylene group may be optionally substituted by one or more C2-6 alkenyl, NRx, O or CO groups, such as —(CH2)2—O—(CH2)2—NH—CO—C(═CH2)—CH2—N(Me)2, —(CH2)5—NH—CO—CH═CH—CH2—N(Me)2, —(CH2)5—NH—CO—C(═CH2)—CH2—N(Me)2, —(CH2)4—NHCOCH═CH—CH2—NMe2, or —CH2—CHF—(CH2)3—NHCO—CH═CHCH2NMe2, —(CH2)3—CHF—CH2NHCO—CH═CHCH2NMe2, or —(CH2)3—CF2—CH2NHCO—CH═CHCH2NMe2.
In a further embodiment, —X—R2 represents a —C1-C10 alkylene-NRxRy group, wherein said alkylene group may be optionally substituted by one or more C2-6 alkenyl, NRx, O or CO groups, such as —(CH2)2—O—(CH2)2—NH—CO—C(═CH2)—CH2—N(Me)2, —(CH2)5—NH—CO—CH═CH—CH2—N(Me)2 or —(CH2)5—NH—CO—C(═CH2)—CH2—N(Me)2.
In one embodiment, R3 represents hydrogen or C1-6 alkyl (such as methyl or ethyl). In a further embodiment, R3 represents hydrogen or C1-6 alkyl (such as methyl). In a yet further embodiment, R3 represents hydrogen or methyl. In a yet further embodiment, R3 represents C1-6 alkyl (such as methyl). In a still yet further embodiment, R3 represents methyl.
In one embodiment, R4 represents:
In a further embodiment, R4 represents C1-6 alkyl (such as methyl). In a yet further embodiment, R4 represents methyl.
In one embodiment, R5 represents:
In one embodiment, R5 represents hydrogen.
In one embodiment, R6 represents:
In a further embodiment, R6 represents haloC1-6 alkyl (such as trifluoromethyl). In a yet further embodiment, R6 represents trifluoromethyl.
In one embodiment, R7 represents:
In a further embodiment, R7 represents hydrogen.
In one particular embodiment, R4 represents methyl, R5 represents hydrogen, R6 represents trifluoromethyl, and R7 represents hydrogen. Thus, in one embodiment, the compound of formula (I) is a compound of formula (I)a:
or a tautomeric or a stereochemically isomeric form, a pharmaceutically acceptable salt or a solvate thereof, wherein n, R1, X, R2 and R3 are as defined herein.
In one embodiment, Rx and Ry independently represent hydrogen or C1-6 alkyl (such as methyl or ethyl). In a further embodiment, Rx and Ry independently represent C1-6 alkyl (such as methyl). In a further embodiment, both of Rx and Ry represent C1-6 alkyl (such as methyl).
In one embodiment, the invention provides a compound of formula (I) which is the free base of a compound of Examples 1-332 or a pharmaceutically acceptable salt or solvate thereof.
In a further embodiment, the invention provides a compound of formula (I) which is the free base of a compound of Examples 1-227 or a pharmaceutically acceptable salt or solvate thereof.
A reference to a compound of the formula (I) and sub-groups thereof also includes ionic forms, salts, solvates, isomers (including geometric and stereochemical isomers), tautomers, N-oxides, esters, prodrugs, isotopes and protected forms thereof, for example, as discussed below; preferably, the salts or tautomers or isomers or N-oxides or solvates thereof; and more preferably, the salts or tautomers or N-oxides or solvates thereof, even more preferably the salts or tautomers or solvates thereof. Hereinafter, compounds and their ionic forms, salts, solvates, isomers (including geometric and stereochemical isomers), tautomers, N-oxides, esters, prodrugs, isotopes and protected forms thereof as defined in any aspect of the invention (except intermediate compounds in chemical processes) are referred to as “compounds of the invention”.
Certain compounds of the formula (I) can exist in the form of salts, for example acid addition salts or, in certain cases salts of organic and inorganic bases such as carboxylate, sulfonate and phosphate salts. All such salts are within the scope of this invention, and references to compounds of the formula (I) include the salt forms of the compounds.
The salts of the present invention can be synthesized from the parent compound that contains a basic or acidic moiety by conventional chemical methods such as methods described in Pharmaceutical Salts: Properties, Selection, and Use, P. Heinrich Stahl (Editor), Camille G. Wermuth (Editor), ISBN: 3-90639-026-8, Hardcover, 388 pages, August 2002. Generally, such salts can be prepared by reacting the free acid or base forms of these compounds with the appropriate base or acid in water or in an organic solvent, or in a mixture of the two; generally, non-aqueous media such as ether, ethyl acetate, ethanol, isopropanol, or acetonitrile are used.
Acid addition salts (mono- or di-salts) may be formed with a wide variety of acids, both inorganic and organic. Examples of acid addition salts include mono- or di-salts formed with an acid selected from the group consisting of acetic, 2,2-dichloroacetic, adipic, alginic, ascorbic (e.g. L-ascorbic), L-aspartic, benzenesulfonic, benzoic, 4-acetamidobenzoic, butanoic, (+) camphoric, camphor-sulfonic, (+)-(1S)-camphor-10-sulfonic, capric, caproic, caprylic, cinnamic, citric, cyclamic, dodecylsulfuric, ethane-1,2-disulfonic, ethanesulfonic, 2-hydroxyethanesulfonic, formic, fumaric, galactaric, gentisic, glucoheptonic, D-gluconic, glucuronic (e.g. D-glucuronic), glutamic (e.g. L-glutamic), α-oxoglutaric, glycolic, hippuric, hydrohalic acids (e.g. hydrobromic, hydrochloric, hydriodic), isethionic, lactic (e.g. (+)-L-lactic, (±)-DL-lactic), lactobionic, maleic, malic, (−)-L-malic, malonic, (±)-DL-mandelic, methanesulfonic, naphthalene-2-sulfonic, naphthalene-1,5-disulfonic, 1-hydroxy-2-naphthoic, nicotinic, nitric, oleic, orotic, oxalic, palmitic, pamoic, phosphoric, propionic, pyruvic, L-pyroglutamic, salicylic, 4-amino-salicylic, sebacic, stearic, succinic, sulfuric, tannic, (+)-L-tartaric, thiocyanic, p-toluenesulfonic, undecylenic and valeric acids, as well as acylated amino acids and cation exchange resins.
One particular group of salts consists of salts formed from acetic, hydrochloric, hydriodic, phosphoric, nitric, sulfuric, citric, lactic, succinic, maleic, malic, isethionic, fumaric, benzenesulfonic, toluenesulfonic, methanesulfonic (mesylate), ethanesulfonic, naphthalenesulfonic, valeric, acetic, propanoic, butanoic, malonic, glucuronic and lactobionic acids. One particular salt is the hydrochloride salt.
Where the compounds of the formula (I) contain an amine function, these may form quaternary ammonium salts, for example by reaction with an alkylating agent according to methods well known to the skilled person. Such quaternary ammonium compounds are within the scope of formula (I).
The compounds of the invention may exist as mono- or di-salts depending upon the pKa of the acid from which the salt is formed.
It will be appreciated that for use in medicine the salts of the compounds of formula (I) should be pharmaceutically acceptable. Suitable pharmaceutically acceptable salts will be apparent to those skilled in the art. Pharmaceutically acceptable salts include those described by Berge, Bighley and Monkhouse, J. Pharm. Sci. 1977, 66, pp. 1-19. Such pharmaceutically acceptable salts include acid addition salts formed with inorganic acids e.g. hydrochloric, hydrobromic, sulfuric, nitric or phosphoric acid and organic acids e.g. succinic, maleic, acetic, fumaric, citric, tartaric, benzoic, p-toluenesulfonic, methanesulfonic or naphthalenesulfonic acid. Other salts e.g. oxalates or formates may be used, for example in the isolation of compounds of formula (I) and are included within the scope of this invention. However, salts that are not pharmaceutically acceptable may also be prepared as intermediate forms which may then be converted into pharmaceutically acceptable salts. Such non-pharmaceutically acceptable salts forms, which may be useful, for example, in the purification or separation of the compounds of the invention, also form part of the invention.
Certain of the compounds of formula (I) may form acid addition salts with one or more equivalents of the acid. The present invention includes within its scope all possible stoichiometric and non-stoichiometric forms.
Those skilled in the art of organic chemistry will appreciate that many organic compounds can form complexes with solvents in which they are reacted or from which they are precipitated or crystallized. These complexes are known as “solvates”. For example, a complex with water is known as a “hydrate”. Pharmaceutically acceptable solvates of the compound of the invention are within the scope of the invention. In one embodiment, the pharmaceutically acceptable solvates of the compounds of the invention include the hydrate thereof.
In one embodiment, said crystalline form of the compounds of formula (I) is a cocrystal or coformer. Such a cocrystal or coformer may be prepared using water-soluble molecules such as saccharin, caffeine, nicotinamide or carboxylic acids. Coformers may be prepared as described in Emami S et al (2018) BioImpacts 8(4), 305-320, the techniques of which are herein incorporated by reference.
It will be understood that the invention includes pharmaceutically acceptable derivatives of compounds of formula (I) and that these are included within the scope of the invention.
As used herein “pharmaceutically acceptable derivative” includes any pharmaceutically acceptable ester or salt of such ester of a compound of formula (I) which, upon administration to the recipient is capable of providing (directly or indirectly) a compound of formula (I) or an active metabolite or residue thereof.
Compounds of the formula (I) containing an amine function may also form N-oxides. A reference herein to a compound of the formula (I) that contains an amine function also includes the N-oxide.
Where a compound contains several amine functions, one or more than one nitrogen atom may be oxidised to form an N-oxide. Particular examples of N-oxides are the N-oxides of a tertiary amine or a nitrogen atom of a nitrogen-containing heterocycle.
N-Oxides can be formed by treatment of the corresponding amine with an oxidizing agent such as hydrogen peroxide or a per-acid (e.g. a peroxycarboxylic acid), see for example Advanced Organic Chemistry, by Jerry March, 4th Edition, Wiley Interscience. More particularly, N-oxides can be made by the procedure of L. W. Deady (Syn. Commun. 1977, 7, 509-514) in which the amine compound is reacted with m-chloroperoxybenzoic acid (mCPBA), for example, in an inert solvent such as dichloromethane.
It will be appreciated by those skilled in the art that certain protected derivatives of compounds of formula (I), which may be made prior to a final deprotection stage, may not possess pharmacological activity as such, but may, in certain instances, be administered orally or parenterally and thereafter metabolised in the body to form compounds of the invention which are pharmacologically active. Such derivatives may therefore be described as “prodrugs”. All such prodrugs of compounds of the invention are included within the scope of the invention. Examples of pro-drug functionality suitable for the compounds of the present invention are described in Drugs of Today, 19, 9, 1983, 499-538 and in Topics in Chemistry, Chapter 31, pp. 306-316 and in “Design of Prodrugs” by H. Bundgaard, Elsevier, 1985, Chapter 1 (the disclosures in which documents are incorporated herein by reference). It will further be appreciated by those skilled in the art, that certain moieties, known to those skilled in the art as “pro-moieties”, for example as described by H. Bundgaard in “Design of Prodrugs” (the disclosure in which document is incorporated herein by reference) may be placed on appropriate functionalities when such functionalities are present within compounds of the invention.
Also included within the scope of the compound and various salts of the invention are polymorphs thereof.
Where chiral centres are present in compounds of formula (I), the present invention includes within its scope all possible enantiomers and diastereoisomers, including mixtures thereof. The different isomeric forms may be separated or resolved one from the other by conventional methods, or any given isomer may be obtained by conventional synthetic methods or by stereospecific or asymmetric syntheses. The invention also extends to any tautomeric forms or mixtures thereof.
The subject invention also includes all pharmaceutically acceptable isotopically-labelled compounds which are identical to those recited in formula (I) but for the fact that one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number most commonly found in nature.
Examples of isotopes suitable for inclusion in the compounds of the invention comprise isotopes of hydrogen, such as 2H (D) and 3H (T), carbon, such as 11C, 13C and 14C, chlorine, such as 36Cl, fluorine, such as 18F, iodine, such as 123I, 125I and 131I, nitrogen, such as 13N and 15N, oxygen, such as 15O, 17O and 18O, phosphorus, such as 32P, and sulfur, such as 35S.
Certain isotopically-labelled compounds of formula (I), for example, those incorporating a radioactive isotope, are useful in drug and/or substrate tissue distribution studies. The compounds of formula (I) can also have valuable diagnostic properties in that they can be used for detecting or identifying the formation of a complex between a labelled compound and other molecules, peptides, proteins, enzymes or receptors. The detecting or identifying methods can use compounds that are labelled with labelling agents such as radioisotopes, enzymes, fluorescent substances, luminous substances (for example, luminol, luminol derivatives, luciferin, aequorin and luciferase) etc. The radioactive isotopes tritium, i.e. 3H (T), and carbon-14, i.e. 14C, are particularly useful for this purpose in view of their ease of incorporation and ready means of detection.
Substitution with heavier isotopes such as deuterium, i.e. 2H (D), may afford certain therapeutic advantages resulting from greater metabolic stability, for example, increased in vivo half-life or reduced dosage requirements, and hence may be preferred in some circumstances.
Substitution with positron emitting isotopes, such as 11C, 18F, 15O and 13N, can be useful in Positron Emission Topography (PET) studies for examining target occupancy.
Isotopically-labelled compounds of formula (I) can generally be prepared by conventional techniques known to those skilled in the art or by processes analogous to those described in the accompanying Examples and Preparations using appropriate isotopically-labelled reagents in place of the non-labelled reagent previously employed.
Since the compounds of formula (I) are intended for use in pharmaceutical compositions it will readily be understood that they are each preferably provided in substantially pure form, for example at least 60% pure, more suitably at least 75% pure and preferably at least 85%, especially at least 98% pure (% are given on a weight for weight basis). Impure preparations of the compounds may be used for preparing the more pure forms used in the pharmaceutical compositions.
According to a further aspect of the present invention there is provided a process for the preparation of compounds of formula (I) and derivatives thereof. The following schemes are examples of synthetic schemes that may be used to synthesise the compounds of the invention. In the following schemes reactive groups can be protected with protecting groups and de-protected according to well established techniques.
According to a further aspect of the invention there is provided a process for preparing a compound of formula (I) as herein defined which comprises:
Process (a) typically comprises a deprotection reaction to prepare the compound of formula (I). For example, when P1 represents Boc, process (a) typically comprises the use of an acid, such as TFA.
Process (b) typically comprises reacting a compound of formula (II) with the compound of formula L1-X—R2 in a suitable solvent, such as DCM in the presence of suitable reagents, such as HATU and DIPEA.
Compounds of formula (II) and (III) may be prepared in accordance with the procedures described herein. For example, compounds of formula (II) may be prepared in accordance with the experimental procedure described in Example 1, compounds of formula (III) may be prepared in accordance with the experimental procedure described in Example 27.
Compounds of formula L1-X—R2 are either known or may be prepared in accordance with known procedures.
A wide range of well known functional group interconversions for process (d) are known by a person skilled in the art for converting a precursor compound to a compound of formula (I) and are described in Advanced Organic Chemistry by Jerry March, 4th Edition, John Wiley & Sons, 1992. For example possible metal catalysed functionalisations such as using organo-tin reagents (the Stille reaction), Grignard reagents and reactions with nitrogen nucleophiles are described in ‘Palladium Reagents and Catalysts’ [Jiro Tsuji, Wiley, ISBN 0-470-85032-9] and Handbook of OrganoPalladium Chemistry for Organic Synthesis [Volume 1, Edited by Ei-ichi Negishi, Wiley, ISBN 0-471-31506-0].
If appropriate, the reactions described herein are followed or preceded by one or more reactions known to the skilled of the art and are performed in an appropriate order to achieve the requisite substitutions on each of the variables defined herein to afford other compounds of formula (I). Non-limiting examples of such reactions whose conditions can be found in the literature include:
It is recognised that the sequence of reactions involving aryl coupling and reduction may be varied. It is also recognised that a wide range of palladium based catalysts are suitable for conducting aryl coupling reactions.
It may also be recognised that isomer separation may occur at any suitable stage in the synthetic sequence. It should be stressed that such chiral separation forms a key aspect of the invention and that such separation may be conducted in accordance with the methodology described herein or may be conducted in accordance with known methodology. It is also recognised that it may be beneficial to temporarily form a protected derivative of an intermediate in the synthesis, for example, a Boc-protected amine, or SEM-protected amide, in order to facilitate chromatographic separation, chiral resolution or to give improved solubility or yields in particular steps.
In many of the reactions described above, it may be necessary to protect one or more groups to prevent reaction from taking place at an undesirable location on the molecule. Examples of protecting groups, and methods of protecting and de-protecting functional groups, can be found in Protective Groups in Organic Synthesis (T. Green and P. Wuts; 4th Edition; John Wiley and Sons, 2007).
A hydroxy group may be protected, for example, as an ether (—OR) or an ester (—OC(═O)R), for example, as: a tert-butyl ether; a tetrahydropyranyl (THP) ether; a benzyl, benzhydryl (diphenylmethyl), or trityl (triphenylmethyl) ether; a trimethylsilyl or tert-butyldimethylsilyl ether; or an acetyl ester (—OC(═O)CH3).
An amine group may be protected, for example, as an amide (—NRCO—R) or a carbamate (—NRCO—OR), for example, as: a methyl amide (—NHCO—CH3); a benzyl carbamate (—NHCO—OCH2C6H5, —NH-Cbz or NH—Z); as a tert-butyl carbamate (—NHCOOC(CH3)3, NH-Boc); a 2-biphenyl-2-propyl carbamate (—NHCO—OC(CH3)2C6H4C6H5, NH-Boc), as a 9-fluorenylmethyl carbamate (—NH-Fmoc), as a 6-nitroveratryl carbamate (—NH-Nvoc), as a 2-trimethylsilylethyl carbamate (—NH-Teoc), as a 2,2,2-trichloroethyl carbamate (—NH-Troc), as an allyl carbamate (—NH-Alloc), or as a 2(-phenylsulfonyl)ethyl carbamate (—NH-Psec).
Other protecting groups for amines, such as cyclic amines and heterocyclic N—H groups, include toluenesulfonyl (tosyl) and methanesulfonyl (mesyl) groups, benzyl groups such as a para-methoxybenzyl (PMB) group and tetrahydropyranyl (THP) groups.
A carboxylic acid group may be protected as an ester for example, as: an C1-7 alkyl ester (e.g. a methyl ester; a tert-butyl ester); a C1-7 haloalkyl ester (e.g. a C1-7 trihaloalkyl ester); a triC1-7 alkylsilyl-C1-7 alkyl ester; or a C5-20 aryl-C1-7 alkyl ester (e.g. a benzyl ester; a nitrobenzyl ester; para-methoxybenzyl ester.
It will be understood by those skilled in the art that certain compounds of the invention can be converted into other compounds of the invention according to standard chemical methods.
Pharmaceutically acceptable salts may be prepared conventionally by reaction with the appropriate acid or acid derivative.
The compounds of the invention, subgroups and examples thereof, are inhibitors of Polθ polymerase activity, and which may be useful in preventing or treating disease states or conditions described herein. In addition, the compounds of the invention, and subgroups thereof, will be useful in preventing or treating diseases or condition mediated by Polθ. References to the preventing or prophylaxis or treatment of a disease state or condition such as cancer include within their scope alleviating or reducing the incidence of cancer.
Thus, for example, it is envisaged that the compounds of the invention will be useful in alleviating or reducing the incidence of cancer.
The compounds of the present invention may be useful for the treatment of the adult population. The compounds of the present invention may be useful for the treatment of the pediatric population.
As a consequence of their inhibition of Polθ, the compounds will be useful in providing a means of disabling the ability of cells to perform MMEJ. It is therefore anticipated that the compounds may prove useful in treating or preventing proliferative disorders such as cancers. In addition, the compounds of the invention may be useful in the treatment of diseases in which there is a disorder associated with cell accumulation.
Without being bound by theory it is expected that the Polθ inhibitors of the present invention will demonstrate certain properties for them to be of particular utility in the therapeutic treatment of certain cancers. For example, in one embodiment, the Polθ inhibitors of the present invention are suitably lethal in BRCA1 and BRCA2 deficient primary and secondary solid tumours, including breast, ovarian, prostate and pancreas.
In a further embodiment, the Polθ inhibitors of the present invention are suitably lethal in a variety of primary and secondary solid tumours which are HRD by mechanisms other than BRCA deficiency, such as those with promoter hypermethylation. In these tumours where no DSB repair pathway may be fully down regulated the Polθi may be given along with another DDR modulator such as a PARP inhibitor, a DNA-PK inhibitor, an ATR inhibitor, an ATM inhibitor, a wee1 inhibitor or a CHK1 inhibitor.
In a further embodiment, the Polθ inhibitors of the present invention are suitably lethal in primary and secondary breast, ovarian, prostate and pancreatic tumours retaining BRCA1 deficiency but which, following or not following exposure to PARPi medication, are resistant to PARPi treatment.
In a further embodiment, the Polθ inhibitors of the present invention suitably increase the ORR including CRR, will delay the onset of PARPi resistance, will increase the time to relapse and DFS, and will increase the OS of HRD (BRCA1/2 deficient and other HRD mechanisms) primary and secondary tumours (breast, ovarian, prostate and pancreas) when given with PARPi treatment programmes.
In a further embodiment, the Polθ inhibitors of the present invention suitably show synthetic sickness and/or synthetic lethality in a variety of tumours with loss of ATM activity (ATM−/−) particularly in the context of WT p53. Tumour types will include around 10% of all solid tumours including gastric, lung, breast, and CRC, along with CLL. Co-medicating with another DDR modifier, such as a DNA-PK inhibitor, PARP inhibitor or ATR inhibitor, may further enhance such activity. Polθ inhibitors will resensitise CLL to classical chemotherapy and chemo-immunotherapy where drug resistance has emerged. Thus, according to a further embodiment, the pharmaceutical composition of the present invention additionally comprises a DNA-PK inhibitor, PARP inhibitor, or ATR inhibitor.
In a further embodiment, the Polθ inhibitors of the present invention suitably show synthetic sickness and/or synthetic lethality in a variety of tumours deficient in the DNA double strand break repair process of non-homologous end-joining (NHEJ-D). Tumour types will include approximately 2-10% of all solid tumours including prostate, pancreatic, cervical, breast, lung, bladder and oesophageal. Co-medicating with another DDR modifier, such as a PARP inhibitor, ATM inhibitor, wee1 inhibitor, CHK inhibitor, or ATR inhibitor, may further enhance such activity. Polθ inhibitors will further sensitise NHEJD cancer cells to DNA DSB inducing chemotherapies and to ionising radiation based therapies. Thus, according to a further embodiment, the pharmaceutical composition of the present invention additionally comprises a PARP inhibitor, ATM inhibitor, wee1 inhibitor, CHK inhibitor, or ATR inhibitor.
In a further embodiment, the Polθ inhibitors of the present invention suitably reduce the DNA replication stress response during the chemotherapy of HR proficient tumours such as ovarian, NSCL and breast tumours over expressing Polθ. This will increase the ORR to treatment and increase OS. Such effects are particularly likely with cytarabine (Ara-C) and hydroxyurea used in a wide variety of leukemias including CML, and the management of squamous cell carcinomas.
In a further embodiment, the Polθ inhibitors of the present invention suitably selectively sensitise solid tumours to radiotherapy, including EBRT and brachytherapy, with little or no sensitisation of normal tissues. In a fractionated curative-intent setting this will increase loco-regional control driving increased survival. This will be particularly evident in the management of NSCLC, SCCH&N, rectal cancer, prostate cancer and pancreatic cancer.
In a further embodiment, the Polθ inhibitors of the present invention suitably show synthetic sickness and/or synthetic lethality in PTEN deleted tumours such as CaP, with or without comedication with a PARPi. Furthermore, such tumours will exhibit exquisite sensitivity to radiotherapy both by dint of the PTEN deletion as well as the Polθ inhibitor induced radiosensitivity.
In a further embodiment, the Polθ inhibitors of the present invention suitably suppress TLS polymerase activity, sensitising primary and secondary solid tumours (e.g. breast, lung, ovarian, CRC) to drugs (e.g. cisplatin, mitomycin and cyclophosphamide) as well as reducing the acquisition of drug-induced mutations implicated in tumour resistance leading to prolongation of remission and increased TTR.
In a further embodiment, the Polθ inhibitors of the present invention suitably resensitise BCR-ABL-positive CML which is has developed imatinib resistance, as well as other solid tumours with elevated ligase IIIα levels, reduced ligase IV levels and increased dependence upon altEJ DSB repair.
In a further embodiment, the Polθ inhibitors of the present invention suitably show synthetic sickness and/or synthetic lethality in aromatase inhibitor resistant ER− primary and secondary breast cancers, again showing elevated ligase IIIα levels, reduced ligase IV levels and increased dependence upon altEJ DSB repair.
According to a further aspect of the invention there is a provided a compound of formula (I) as defined herein for use in the treatment of tumours characterised by a deficiency in homologous recombination (HRD).
It will be appreciated that references herein to “deficiency in homologous recombination (HRD)” refer to any genetic variation which results in a deficiency or loss of function of the resultant homologous recombination gene. Examples of said genetic variation include mutations (e.g. point mutations), substitutions, deletions, single nucleotide polymorphisms (SNPs), haplotypes, chromosome abnormalities, Copy Number Variation (CNV), epigenetics, DNA inversions, reduction in expression and mis-localisation.
In one embodiment, said homologous recombination genes are selected from any of: ATM, ATR, BRCA1, BRCA2, BARD1, RAD51C, RAD50, CHEK1, CHEK2, FANCA, FANCB, FANCC, FANCD2, FANCE, FANCF, FANCG, FANCI, FANCL, FANCM, PALB2 (FANCN), FANCP (BTBD12), ERCC4 (FANCQ), PTEN, CDK12, MRE11, NBS1, NBN, CLASPIN, BLM, WRN, SMARCA2, SMARCA4, LIG1, RPA1, RPA2, BRIP1 and PTEN.
It will be appreciated that references herein to “non-homologous end-joining deficiency (NHEJD)” refer to any genetic variation which results in a deficiency or loss of function of the resultant homologous recombination gene. Examples of said genetic variation include mutations (e.g. point mutations), substitutions, deletions, single nucleotide polymorphisms (SNPs), haplotypes, chromosome abnormalities, Copy Number Variation (CNV), epigenetics, DNA inversions, reduction in expression and mis-localisation.
In one embodiment, said non-homologous end-joining genes are selected from any one or more of: LIG4, NHEJ1, POLL, POLM, PRKDC, XRCC4, XRCC5, XRCC6, and DCLRE1C.
According to a further aspect of the invention there is a provided a compound of formula (I) as defined herein for use in the treatment of tumours which overexpress Polθ.
According to a further aspect of the invention there is a provided a compound of formula (I) as defined herein for use in the treatment of tumours which have elevated ligase IIIα levels, reduced ligase IV levels and increased dependence upon altEJ DSB repair.
Examples of cancers (and their benign counterparts) which may be treated (or inhibited) include, but are not limited to tumours of epithelial origin (adenomas and carcinomas of various types including adenocarcinomas, squamous carcinomas, transitional cell carcinomas and other carcinomas) such as carcinomas of the bladder and urinary tract, breast, gastrointestinal tract (including the esophagus, stomach (gastric), small intestine, colon, rectum and anus), liver (hepatocellular carcinoma), gall bladder and biliary system, exocrine pancreas, kidney, lung (for example adenocarcinomas, small cell lung carcinomas, non-small cell lung carcinomas, bronchioalveolar carcinomas and mesotheliomas), head and neck (for example cancers of the tongue, buccal cavity, larynx, pharynx, nasopharynx, tonsil, salivary glands, nasal cavity and paranasal sinuses), ovary, fallopian tubes, peritoneum, vagina, vulva, penis, cervix, myometrium, endometrium, thyroid (for example thyroid follicular carcinoma), adrenal, prostate, skin and adnexae (for example melanoma, basal cell carcinoma, squamous cell carcinoma, keratoacanthoma, dysplastic naevus); haematological malignancies (i.e. leukemias, lymphomas) and premalignant haematological disorders and disorders of borderline malignancy including haematological malignancies and related conditions of lymphoid lineage (for example acute lymphocytic leukemia [ALL], chronic lymphocytic leukemia [CLL], B-cell lymphomas such as diffuse large B-cell lymphoma [DLBCL], follicular lymphoma, Burkitt's lymphoma, mantle cell lymphoma, MALT lymphoma, T-cell lymphomas and leukaemias, natural killer [NK] cell lymphomas, Hodgkin's lymphomas, hairy cell leukaemia, monoclonal gammopathy of uncertain significance, plasmacytoma, multiple myeloma, and post-transplant lymphoproliferative disorders), and haematological malignancies and related conditions of myeloid lineage (for example acute myelogenous leukemia [AML], chronic myelogenous leukemia [CML], chronic myelomonocytic leukemia [CMML], hypereosinophilic syndrome, myeloproliferative disorders such as polycythaemia vera, essential thrombocythaemia and primary myelofibrosis, myeloproliferative syndrome, myelodysplastic syndrome, and promyelocytic leukemia); tumours of mesenchymal origin, for example sarcomas of soft tissue, bone or cartilage such as osteosarcomas, fibrosarcomas, chondrosarcomas, rhabdomyosarcomas, leiomyosarcomas, liposarcomas, angiosarcomas, Kaposi's sarcoma, Ewing's sarcoma, synovial sarcomas, epithelioid sarcomas, gastrointestinal stromal tumours, benign and malignant histiocytomas, and dermatofibrosarcoma protuberans; tumours of the central or peripheral nervous system (for example astrocytomas, gliomas and glioblastomas, meningiomas, ependymomas, pineal tumours and schwannomas); endocrine tumours (for example pituitary tumours, adrenal tumours, islet cell tumours, parathyroid tumours, carcinoid tumours and medullary carcinoma of the thyroid); ocular and adnexal tumours (for example retinoblastoma); germ cell and trophoblastic tumours (for example teratomas, seminomas, dysgerminomas, hydatidiform moles and choriocarcinomas); and paediatric and embryonal tumours (for example medulloblastoma, neuroblastoma, Wilms tumour, and primitive neuroectodermal tumours); or syndromes, congenital or otherwise, which leave the patient susceptible to malignancy (for example Xeroderma Pigmentosum).
Many diseases are characterized by persistent and unregulated angiogenesis. Chronic proliferative diseases are often accompanied by profound angiogenesis, which can contribute to or maintain an inflammatory and/or proliferative state, or which leads to tissue destruction through the invasive proliferation of blood vessels. Tumour growth and metastasis have been found to be angiogenesis-dependent. Compounds of the invention may therefore be useful in preventing and disrupting initiation of tumour angiogenesis. In particular, the compounds of the invention may be useful in the treatment of metastasis and metastatic cancers.
Metastasis or metastatic disease is the spread of a disease from one organ or part to another non-adjacent organ or part. The cancers which can be treated by the compounds of the invention include primary tumours (i.e. cancer cells at the originating site), local invasion (cancer cells which penetrate and infiltrate surrounding normal tissues in the local area), and metastatic (or secondary) tumours ie. tumours that have formed from malignant cells which have circulated through the bloodstream (haematogenous spread) or via lymphatics or across body cavities (trans-coelomic) to other sites and tissues in the body.
Particular cancers include hepatocellular carcinoma, melanoma, oesophageal, renal, colon, colorectal, lung e.g. mesothelioma or lung adenocarcinoma, breast, bladder, gastrointestinal, ovarian and prostate cancers.
A further aspect provides the use of a compound for the manufacture of a medicament for the treatment of a disease or condition as described herein, in particular cancer.
The compounds may also be useful in the treatment of tumour growth, pathogenesis, resistance to chemo- and radio-therapy by sensitising cells to chemotherapy and as an anti-metastatic agent.
The potency of the compounds of the invention as inhibitors of Polθ can be measured using the biological and biophysical assays set forth in the examples herein and the level of affinity exhibited by a given compound can be defined in terms of the IC50 value. Particular compounds of the present invention are compounds having an IC50 value of less than 1 μM, more particularly less than 0.1 μM.
A role for the loss of Polθ enhancing the efficacy of CRISPR mediated gene editing has been described in WO 2017/062754. Thus, Polθ inhibitory compounds are likely to be useful in enhancing the efficiency of CRISPR based editing methodologies and/or CRISPR based editing therapeutics. Furthermore, compound mediated Polθ inhibition is likely to reduce the frequency of random integration events and thus provide a route to ameliorate any safety concerns of CRISPR mediated technology. Thus, according to a further aspect of the invention, there is provided the use of a compound of formula (I) as defined herein in a CRISPR based editing methodology and/or CRISPR based editing therapeutics, such as the enhancement of efficiency of CRISPR based editing methodology and/or CRISPR based editing therapeutics.
While it is possible for the active compound to be administered alone, it is preferable to present it as a pharmaceutical composition (e.g. formulation). In one embodiment this is a sterile pharmaceutical composition.
Thus, the present invention further provides pharmaceutical compositions, as defined above, and methods of making a pharmaceutical composition comprising (e.g admixing) at least one compound of formula (I) (and sub-groups thereof as defined herein), together with one or more pharmaceutically acceptable excipients and optionally other therapeutic or prophylactic agents, as described herein.
The pharmaceutically acceptable excipient(s) can be selected from, for example, carriers (e.g. a solid, liquid or semi-solid carrier), adjuvants, diluents, fillers or bulking agents, granulating agents, coating agents, release-controlling agents, binding agents, disintegrants, lubricating agents, preservatives, antioxidants, buffering agents, suspending agents, thickening agents, flavouring agents, sweeteners, taste masking agents, stabilisers or any other excipients conventionally used in pharmaceutical compositions. Examples of excipients for various types of pharmaceutical compositions are set out in more detail below.
The term “pharmaceutically acceptable” as used herein pertains to compounds, materials, compositions, and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of a subject (e.g. human) without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio. Each carrier, excipient, etc. must also be “acceptable” in the sense of being compatible with the other ingredients of the formulation.
Pharmaceutical compositions containing compounds of the formula (I) can be formulated in accordance with known techniques, see for example, Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, PA, USA.
The pharmaceutical compositions can be in any form suitable for oral, parenteral, topical, intranasal, intrabronchial, sublingual, ophthalmic, otic, rectal, intra-vaginal, or transdermal administration. Where the compositions are intended for parenteral administration, they can be formulated for intravenous, intramuscular, intraperitoneal, subcutaneous administration or for direct delivery into a target organ or tissue by injection, infusion or other means of delivery. The delivery can be by bolus injection, short term infusion or longer term infusion and can be via passive delivery or through the utilisation of a suitable infusion pump or syringe driver.
Pharmaceutical formulations adapted for parenteral administration include aqueous and non-aqueous sterile injection solutions which may contain anti-oxidants, buffers, bacteriostats, co-solvents, surface active agents, organic solvent mixtures, cyclodextrin complexation agents, emulsifying agents (for forming and stabilizing emulsion formulations), liposome components for forming liposomes, gellable polymers for forming polymeric gels, lyophilisation protectants and combinations of agents for, inter alia, stabilising the active ingredient in a soluble form and rendering the formulation isotonic with the blood of the intended recipient. Pharmaceutical formulations for parenteral administration may also take the form of aqueous and non-aqueous sterile suspensions which may include suspending agents and thickening agents (R. G. Strickly, Solubilizing Excipients in oral and injectable formulations, Pharmaceutical Research, Vol 21(2) 2004, p 201-230).
The formulations may be presented in unit-dose or multi-dose containers, for example sealed ampoules, vials and prefilled syringes, and may be stored in a freeze-dried (lyophilised) condition requiring only the addition of the sterile liquid carrier, for example water for injections, immediately prior to use. In one embodiment, the formulation is provided as an active pharmaceutical ingredient in a bottle for subsequent reconstitution using an appropriate diluent.
The pharmaceutical formulation can be prepared by lyophilising a compound of formula (I), or sub-groups thereof. Lyophilisation refers to the procedure of freeze-drying a composition. Freeze-drying and lyophilisation are therefore used herein as synonyms.
Extemporaneous injection solutions and suspensions may be prepared from sterile powders, granules and tablets.
Pharmaceutical compositions of the present invention for parenteral injection can also comprise pharmaceutically acceptable sterile aqueous or non-aqueous solutions, dispersions, suspensions or emulsions as well as sterile powders for reconstitution into sterile injectable solutions or dispersions just prior to use.
Examples of suitable aqueous and nonaqueous carriers, diluents, solvents or vehicles include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol, and the like), carboxymethylcellulose and suitable mixtures thereof, vegetable oils (such as sunflower oil, safflower oil, corn oil or olive oil), and injectable organic esters such as ethyl oleate. Proper fluidity can be maintained, for example, by the use of thickening or coating materials such as lecithin, by the maintenance of the required particle size in the case of dispersions, and by the use of surfactants.
The compositions of the present invention may also contain adjuvants such as preservatives, wetting agents, emulsifying agents, and dispersing agents. Prevention of the action of microorganisms may be ensured by the inclusion of various antibacterial and antifungal agents, for example, paraben, chlorobutanol, phenol, sorbic acid, and the like. It may also be desirable to include agents to adjust tonicity such as sugars, sodium chloride, and the like. Prolonged absorption of the injectable pharmaceutical form may be brought about by the inclusion of agents which delay absorption such as aluminum monostearate and gelatin.
In one particular embodiment of the invention, the pharmaceutical composition is in a form suitable for i.v. administration, for example by injection or infusion. For intravenous administration, the solution can be dosed as is, or can be injected into an infusion bag (containing a pharmaceutically acceptable excipient, such as 0.9% saline or 5% dextrose), before administration.
In another particular embodiment, the pharmaceutical composition is in a form suitable for sub-cutaneous (s.c.) administration.
Pharmaceutical dosage forms suitable for oral administration include tablets (coated or uncoated), capsules (hard or soft shell), caplets, pills, lozenges, syrups, solutions, powders, granules, elixirs and suspensions, sublingual tablets, wafers or patches such as buccal patches.
Thus, tablet compositions can contain a unit dosage of active compound together with an inert diluent or carrier such as a sugar or sugar alcohol, eg; lactose, sucrose, sorbitol or mannitol; and/or a non-sugar derived diluent such as sodium carbonate, calcium phosphate, calcium carbonate, or a cellulose or derivative thereof such as microcrystalline cellulose (MCC), methyl cellulose, ethyl cellulose, hydroxypropyl methyl cellulose, and starches such as corn starch. Tablets may also contain such standard ingredients as binding and granulating agents such as polyvinylpyrrolidone, disintegrants (e.g. swellable crosslinked polymers such as crosslinked carboxymethylcellulose), lubricating agents (e.g. stearates), preservatives (e.g. parabens), antioxidants (e.g. BHT), buffering agents (for example phosphate or citrate buffers), and effervescent agents such as citrate/bicarbonate mixtures. Such excipients are well known and do not need to be discussed in detail here.
Tablets may be designed to release the drug either upon contact with stomach fluids (immediate release tablets) or to release in a controlled manner (controlled release tablets) over a prolonged period of time or with a specific region of the GI tract.
Capsule formulations may be of the hard gelatin or soft gelatin variety and can contain the active component in solid, semi-solid, or liquid form. Gelatin capsules can be formed from animal gelatin or synthetic or plant derived equivalents thereof.
The solid dosage forms (eg; tablets, capsules etc.) can be coated or un-coated. Coatings may act either as a protective film (e.g. a polymer, wax or varnish) or as a mechanism for controlling drug release or for aesthetic or identification purposes. The coating (e.g. a Eudragit™ type polymer) can be designed to release the active component at a desired location within the gastro-intestinal tract. Thus, the coating can be selected so as to degrade under certain pH conditions within the gastrointestinal tract, thereby selectively release the compound in the stomach or in the ileum, duodenum, jejenum or colon.
Instead of, or in addition to, a coating, the drug can be presented in a solid matrix comprising a release controlling agent, for example a release delaying agent which may be adapted to release the compound in a controlled manner in the gastrointestinal tract. Alternatively the drug can be presented in a polymer coating e.g. a polymethacrylate polymer coating, which may be adapted to selectively release the compound under conditions of varying acidity or alkalinity in the gastrointestinal tract. Alternatively, the matrix material or release retarding coating can take the form of an erodible polymer (e.g. a maleic anhydride polymer) which is substantially continuously eroded as the dosage form passes through the gastrointestinal tract. In another alternative, the coating can be designed to disintegrate under microbial action in the gut. As a further alternative, the active compound can be formulated in a delivery system that provides osmotic control of the release of the compound. Osmotic release and other delayed release or sustained release formulations (for example formulations based on ion exchange resins) may be prepared in accordance with methods well known to those skilled in the art.
The compound of formula (I) may be formulated with a carrier and administered in the form of nanoparticles, the increased surface area of the nanoparticles assisting their absorption. In addition, nanoparticles offer the possibility of direct penetration into the cell. Nanoparticle drug delivery systems are described in “Nanoparticle Technology for Drug Delivery”, edited by Ram B Gupta and Uday B. Kompella, Informa Healthcare, ISBN 9781574448573, published 13th Mar. 2006. Nanoparticles for drug delivery are also described in J. Control. Release, 2003, 91 (1-2), 167-172, and in Sinha et al., Mol. Cancer Ther. August 1, (2006) 5, 1909.
The pharmaceutical compositions typically comprise from approximately 1% (w/w) to approximately 95% (w/w) active ingredient and from 99% (w/w) to 5% (w/w) of a pharmaceutically acceptable excipient or combination of excipients. Particularly, the compositions comprise from approximately 20% (w/w) to approximately 90%,% (w/w) active ingredient and from 80% (w/w) to 10% of a pharmaceutically acceptable excipient or combination of excipients. The pharmaceutical compositions comprise from approximately 1% to approximately 95%, particularly from approximately 20% to approximately 90%, active ingredient. Pharmaceutical compositions according to the invention may be, for example, in unit dose form, such as in the form of ampoules, vials, suppositories, pre-filled syringes, dragées, tablets or capsules.
The pharmaceutically acceptable excipient(s) can be selected according to the desired physical form of the formulation and can, for example, be selected from diluents (e.g solid diluents such as fillers or bulking agents; and liquid diluents such as solvents and co-solvents), disintegrants, buffering agents, lubricants, flow aids, release controlling (e.g. release retarding or delaying polymers or waxes) agents, binders, granulating agents, pigments, plasticizers, antioxidants, preservatives, flavouring agents, taste masking agents, tonicity adjusting agents and coating agents.
The skilled person will have the expertise to select the appropriate amounts of ingredients for use in the formulations. For example tablets and capsules typically contain 0-20% disintegrants, 0-5% lubricants, 0-5% flow aids and/or 0-99% (w/w) fillers/or bulking agents (depending on drug dose). They may also contain 0-10% (w/w) polymer binders, 0-5% (w/w) antioxidants, 0-5% (w/w) pigments. Slow release tablets would in addition contain 0-99% (w/w) release-controlling (e.g. delaying) polymers (depending on dose). The film coats of the tablet or capsule typically contain 0-10% (w/w) polymers, 0-3% (w/w) pigments, and/or 0-2% (w/w) plasticizers.
Parenteral formulations typically contain 0-20% (w/w) buffers, 0-50% (w/w) cosolvents, and/or 0-99% (w/w) Water for Injection (WFI) (depending on dose and if freeze dried). Formulations for intramuscular depots may also contain 0-99% (w/w) oils.
Pharmaceutical compositions for oral administration can be obtained by combining the active ingredient with solid carriers, if desired granulating a resulting mixture, and processing the mixture, if desired or necessary, after the addition of appropriate excipients, into tablets, dragee cores or capsules. It is also possible for them to be incorporated into a polymer or waxy matrix that allow the active ingredients to diffuse or be released in measured amounts.
The compounds of the invention can also be formulated as solid dispersions. Solid dispersions are homogeneous extremely fine disperse phases of two or more solids. Solid solutions (molecularly disperse systems), one type of solid dispersion, are well known for use in pharmaceutical technology (see (Chiou and Riegelman, J. Pharm. Sci., 60, 1281-1300 (1971)) and are useful in increasing dissolution rates and increasing the bioavailability of poorly water-soluble drugs.
This invention also provides solid dosage forms comprising the solid solution described above. Solid dosage forms include tablets, capsules, chewable tablets and dispersible or effervescent tablets. Known excipients can be blended with the solid solution to provide the desired dosage form. For example, a capsule can contain the solid solution blended with (a) a disintegrant and a lubricant, or (b) a disintegrant, a lubricant and a surfactant. In addition a capsule can contain a bulking agent, such as lactose or microcrystalline cellulose. A tablet can contain the solid solution blended with at least one disintegrant, a lubricant, a surfactant, a bulking agent and a glidant. A chewable tablet can contain the solid solution blended with a bulking agent, a lubricant, and if desired an additional sweetening agent (such as an artificial sweetener), and suitable flavours. Solid solutions may also be formed by spraying solutions of drug and a suitable polymer onto the surface of inert carriers such as sugar beads (‘non-pareils’). These beads can subsequently be filled into capsules or compressed into tablets.
The pharmaceutical formulations may be presented to a patient in “patient packs” containing an entire course of treatment in a single package, usually a blister pack. Patient packs have an advantage over traditional prescriptions, where a pharmacist divides a patient's supply of a pharmaceutical from a bulk supply, in that the patient always has access to the package insert contained in the patient pack, normally missing in patient prescriptions. The inclusion of a package insert has been shown to improve patient compliance with the physician's instructions.
Compositions for topical use and nasal delivery include ointments, creams, sprays, patches, gels, liquid drops and inserts (for example intraocular inserts). Such compositions can be formulated in accordance with known methods.
Examples of formulations for rectal or intra-vaginal administration include pessaries and suppositories which may be, for example, formed from a shaped moldable or waxy material containing the active compound. Solutions of the active compound may also be used for rectal administration.
Compositions for administration by inhalation may take the form of inhalable powder compositions or liquid or powder sprays, and can be administrated in standard form using powder inhaler devices or aerosol dispensing devices. Such devices are well known. For administration by inhalation, the powdered formulations typically comprise the active compound together with an inert solid powdered diluent such as lactose.
The compounds of the formula (I) will generally be presented in unit dosage form and, as such, will typically contain sufficient compound to provide a desired level of biological activity. For example, a formulation may contain from 1 nanogram to 2 grams of active ingredient, e.g. from 1 nanogram to 2 milligrams of active ingredient. Within these ranges, particular sub-ranges of compound are 0.1 milligrams to 2 grams of active ingredient (more usually from 10 milligrams to 1 gram, e.g. 50 milligrams to 500 milligrams), or 1 microgram to 20 milligrams (for example 1 microgram to 10 milligrams, e.g. 0.1 milligrams to 2 milligrams of active ingredient).
For oral compositions, a unit dosage form may contain from 1 milligram to 2 grams, more typically 10 milligrams to 1 gram, for example 50 milligrams to 1 gram, e.g. 100 milligrams to 1 gram, of active compound.
The active compound will be administered to a patient in need thereof (for example a human or animal patient) in an amount sufficient to achieve the desired therapeutic effect.
The compounds of the formula (I) and sub-groups as defined herein may be useful in the prophylaxis or treatment of a range of disease states or conditions mediated by Polθ. Thus, according to a further aspect of the invention there is provided a method of treating a disease state or condition mediated by Polθ (e.g. cancer) which comprises administering to a subject in need thereof a compound of formula (I) as described herein. Examples of such disease states and conditions are set out above, and in particular include cancer.
The compounds are generally administered to a subject in need of such administration, for example a human or animal patient, particularly a human.
The compounds will typically be administered in amounts that are therapeutically or prophylactically useful and which generally are non-toxic. However, in certain situations (for example in the case of life threatening diseases), the benefits of administering a compound of the formula (I) may outweigh the disadvantages of any toxic effects or side effects, in which case it may be considered desirable to administer compounds in amounts that are associated with a degree of toxicity.
The compounds may be administered over a prolonged term to maintain beneficial therapeutic effects or may be administered for a short period only. Alternatively they may be administered in a continuous manner or in a manner that provides intermittent dosing (e.g. a pulsatile manner).
A typical daily dose of the compound of formula (I) can be in the range from 100 picograms to 100 milligrams per kilogram of body weight, more typically 5 nanograms to 25 milligrams per kilogram of bodyweight, and more usually 10 nanograms to 15 milligrams per kilogram (e.g. 10 nanograms to 10 milligrams, and more typically 1 microgram per kilogram to 20 milligrams per kilogram, for example 1 microgram to 10 milligrams per kilogram) per kilogram of bodyweight although higher or lower doses may be administered where required. The compound of the formula (I) can be administered on a daily basis or on a repeat basis every 2, or 3, or 4, or 5, or 6, or 7, or 10 or 14, or 21, or 28 days for example.
The compounds of the invention may be administered orally in a range of doses, for example 1 to 1500 mg, 2 to 800 mg, or 5 to 500 mg, e.g. 2 to 200 mg or 10 to 1000 mg, particular examples of doses including 10, 20, 50 and 80 mg. The compound may be administered once or more than once each day. The compound can be administered continuously (i.e. taken every day without a break for the duration of the treatment regimen). Alternatively, the compound can be administered intermittently (i.e. taken continuously for a given period such as a week, then discontinued for a period such as a week and then taken continuously for another period such as a week and so on throughout the duration of the treatment regimen). Examples of treatment regimens involving intermittent administration include regimens wherein administration is in cycles of one week on, one week off; or two weeks on, one week off; or three weeks on, one week off; or two weeks on, two weeks off; or four weeks on two weeks off; or one week on three weeks off—for one or more cycles, e.g. 2, 3, 4, 5, 6, 7, 8, 9 or 10 or more cycles.
In one particular dosing schedule, a patient will be given an infusion of a compound of the formula (I) for periods of one hour daily for up to ten days in particular up to five days for one week, and the treatment repeated at a desired interval such as two to four weeks, in particular every three weeks.
More particularly, a patient may be given an infusion of a compound of the formula (I) for periods of one hour daily for 5 days and the treatment repeated every three weeks.
In another particular dosing schedule, a patient is given an infusion over 30 minutes to 1 hour followed by maintenance infusions of variable duration, for example 1 to 5 hours, e.g. 3 hours.
In a further particular dosing schedule, a patient is given a continuous infusion for a period of 12 hours to 5 days, an in particular a continuous infusion of 24 hours to 72 hours.
In another particular dosing schedule, a patient is given the compound orally once a week.
In another particular dosing schedule, a patient is given the compound orally once-daily for between 7 and 28 days such as 7, 14 or 28 days.
In another particular dosing schedule, a patient is given the compound orally once-daily for 1 day, 2 days, 3 days, 5 days or 1 week followed by the required amount of days off to complete a one or two week cycle.
In another particular dosing schedule, a patient is given the compound orally once-daily for 2 weeks followed by 2 weeks off.
In another particular dosing schedule, a patient is given the compound orally once-daily for 2 weeks followed by 1 week off.
In another particular dosing schedule, a patient is given the compound orally once-daily for 1 week followed by 1 week off.
Ultimately, however, the quantity of compound administered and the type of composition used will be commensurate with the nature of the disease or physiological condition being treated and will be at the discretion of the physician.
It will be appreciated that Polθ inhibitors can be used as a single agent or in combination with other anticancer agents. Combination experiments can be performed, for example, as described in Chou T C, Talalay P. Quantitative analysis of dose-effect relationships: the combined effects of multiple drugs or enzyme inhibitors. Adv Enzyme Regulat 1984; 22: 27-55.
The compounds as defined herein can be administered as the sole therapeutic agent or they can be administered in combination therapy with one of more other compounds (or therapies) for treatment of a particular disease state, for example a neoplastic disease such as a cancer as hereinbefore defined. For the treatment of the above conditions, the compounds of the invention may be advantageously employed in combination with one or more other medicinal agents, more particularly, with other anti-cancer agents or adjuvants (supporting agents in the therapy) in cancer therapy. Examples of other therapeutic agents or treatments that may be administered together (whether concurrently or at different time intervals) with the compounds of the formula (I) include but are not limited to:
Particular examples of anti-cancer agents or adjuvants (or salts thereof), include but are not limited to any of the agents selected from groups (i)-(xlvi), and optionally group (xlvii), below:
In one embodiment the anticancer is selected from recombinant interferons (such as interferon-γ and interferon α) and interleukins (e.g. interleukin 2), for example aldesleukin, denileukin diftitox, interferon alfa 2a, interferon alfa 2b, or peginterferon alfa 2b; interferon-α2 (500 μ/ml) in particular interferon-β; and signal transduction inhibitors such as kinase inhibitors (e.g. EGFR (epithelial growth factor receptor) inhibitors, VEGFR (vascular endothelial growth factor receptor) inhibitors, PDGFR (platelet-derived growth factor receptor) inhibitors, MTKI (multi target kinase inhibitors), Raf inhibitors, mTOR inhibitors for example imatinib mesylate, erlotinib, gefitinib, dasatinib, lapatinib, dovotinib, axitinib, nilotinib, vandetanib, vatalinib, pazopanib, sorafenib, sunitinib, temsirolimus, everolimus (RAD 001), vemurafenib (PLX4032/RG7204), dabrafenib, encorafenib or an IκB kinase inhibitor such as SAR-113945, bardoxolone, BMS-066, BMS-345541, IMD-0354, IMD-2560, or IMD-1041, or MEK inhibitors such as Selumetinib (AZD6244) and Trametinib (GSK121120212), in particular Raf inhibitors (e.g. vemurafenib) or MEK inhibitors (e.g. trametinib).
Each of the compounds present in the combinations of the invention may be given in individually varying dose schedules and via different routes. As such, the posology of each of the two or more agents may differ: each may be administered at the same time or at different times. A person skilled in the art would know through his or her common general knowledge the dosing regimes and combination therapies to use. For example, the compound of the invention may be using in combination with one or more other agents which are administered according to their existing combination regimen. Examples of standard combination regimens are provided below.
The taxane compound is advantageously administered in a dosage of 50 to 400 mg per square meter (mg/m2) of body surface area, for example 75 to 250 mg/m2, particularly for paclitaxel in a dosage of about 175 to 250 mg/m2 and for docetaxel in about 75 to 150 mg/m2 per course of treatment.
The camptothecin compound is advantageously administered in a dosage of 0.1 to 400 mg per square meter (mg/m2) of body surface area, for example 1 to 300 mg/m2, particularly for irinotecan in a dosage of about 100 to 350 mg/m2 and for topotecan in about 1 to 2 mg/m2 per course of treatment.
The anti-tumour podophyllotoxin derivative is advantageously administered in a dosage of 30 to 300 mg per square meter (mg/m2) of body surface area, for example 50 to 250 mg/m2, particularly for etoposide in a dosage of about 35 to 100 mg/m2 and for teniposide in about 50 to 250 mg/m2 per course of treatment.
The anti-tumour vinca alkaloid is advantageously administered in a dosage of 2 to 30 mg per square meter (mg/m2) of body surface area, particularly for vinblastine in a dosage of about 3 to 12 mg/m2, for vincristine in a dosage of about 1 to 2 mg/m2, and for vinorelbine in dosage of about 10 to 30 mg/m2 per course of treatment.
The anti-tumour nucleoside derivative is advantageously administered in a dosage of 200 to 2500 mg per square meter (mg/m2) of body surface area, for example 700 to 1500 mg/m2, particularly for 5-FU in a dosage of 200 to 500 mg/m2, for gemcitabine in a dosage of about 800 to 1200 mg/m2 and for capecitabine in about 1000 to 2500 mg/m2 per course of treatment.
The alkylating agents such as nitrogen mustard or nitrosourea is advantageously administered in a dosage of 100 to 500 mg per square meter (mg/m2) of body surface area, for example 120 to 200 mg/m2, particularly for cyclophosphamide in a dosage of about 100 to 500 mg/m2, for chlorambucil in a dosage of about 0.1 to 0.2 mg/kg, for carmustine in a dosage of about 150 to 200 mg/m2, and for lomustine in a dosage of about 100 to 150 mg/m2 per course of treatment.
The anti-tumour anthracycline derivative is advantageously administered in a dosage of 10 to 75 mg per square meter (mg/m2) of body surface area, for example 15 to
60 mg/m2, particularly for doxorubicin in a dosage of about 40 to 75 mg/m2, for daunorubicin in a dosage of about 25 to 45 mg/m2, and for idarubicin in a dosage of about 10 to 15 mg/m2 per course of treatment.
The antiestrogen agent is advantageously administered in a dosage of about 1 to 100 mg daily depending on the particular agent and the condition being treated. Tamoxifen is advantageously administered orally in a dosage of 5 to 50 mg, particularly 10 to 20 mg twice a day, continuing the therapy for sufficient time to achieve and maintain a therapeutic effect. Toremifene is advantageously administered orally in a dosage of about 60 mg once a day, continuing the therapy for sufficient time to achieve and maintain a therapeutic effect. Anastrozole is advantageously administered orally in a dosage of about 1 mg once a day. Droloxifene is advantageously administered orally in a dosage of about 20-100 mg once a day. Raloxifene is advantageously administered orally in a dosage of about 60 mg once a day. Exemestane is advantageously administered orally in a dosage of about 25 mg once a day.
Antibodies are advantageously administered in a dosage of about 1 to 5 mg per square meter (mg/m2) of body surface area, or as known in the art, if different. Trastuzumab is advantageously administered in a dosage of 1 to 5 mg per square meter (mg/m2) of body surface area, particularly 2 to 4 mg/m2 per course of treatment.
Where the compound of the formula (I) is administered in combination therapy with one, two, three, four or more other therapeutic agents (particularly one or two, more particularly one), the compounds can be administered simultaneously or sequentially. In the latter case, the two or more compounds will be administered within a period and in an amount and manner that is sufficient to ensure that an advantageous or synergistic effect is achieved. When administered sequentially, they can be administered at closely spaced intervals (for example over a period of 5-10 minutes) or at longer intervals (for example 1, 2, 3, 4 or more hours apart, or even longer periods apart where required), the precise dosage regimen being commensurate with the properties of the therapeutic agent(s). These dosages may be administered for example once, twice or more per course of treatment, which may be repeated for example every 7, 14, 21 or 28 days.
In one embodiment is provided a compound of formula (I) for the manufacture of a medicament for use in therapy wherein said compound is used in combination with one, two, three, or four other therapeutic agents. In another embodiment is provided a medicament for treating cancer which comprises a compound of formula (I) wherein said medicament is used in combination with one, two, three, or four other therapeutic agents. The invention further provides use of a compound of formula (I) for the manufacture of a medicament for enhancing or potentiating the response rate in a patient suffering from a cancer where the patient is being treated with one, two, three, or four other therapeutic agents.
It will be appreciated that the particular method and order of administration and the respective dosage amounts and regimes for each component of the combination will depend on the particular other medicinal agent and compound of the present invention being administered, their route of administration, the particular tumour being treated and the particular host being treated. The optimum method and order of administration and the dosage amounts and regime can be readily determined by those skilled in the art using conventional methods and in view of the information set out herein.
The weight ratio of the compound according to the present invention and the one or more other anticancer agent(s) when given as a combination may be determined by the person skilled in the art. Said ratio and the exact dosage and frequency of administration depends on the particular compound according to the invention and the other anticancer agent(s) used, the particular condition being treated, the severity of the condition being treated, the age, weight, gender, diet, time of administration and general physical condition of the particular patient, the mode of administration as well as other medication the individual may be taking, as is well known to those skilled in the art. Furthermore, it is evident that the effective daily amount may be lowered or increased depending on the response of the treated subject and/or depending on the evaluation of the physician prescribing the compounds of the instant invention. A particular weight ratio for the present compound of formula (I) and another anticancer agent may range from 1/10 to 10/1, more in particular from 1/5 to 5/1, even more in particular from 1/3 to 3/1.
The compounds of the invention may also be administered in conjunction with non-chemotherapeutic treatments such as radiotherapy, photodynamic therapy, gene therapy; surgery and controlled diets.
The compounds of the present invention also have therapeutic applications in sensitising tumour cells for radiotherapy and chemotherapy. Hence the compounds of the present invention can be used as “radiosensitizer” and/or “chemosensitizer” or can be given in combination with another “radiosensitizer” and/or “chemosensitizer”. In one embodiment the compound of the invention is for use as chemosensitiser.
The term “radiosensitizer” is defined as a molecule administered to patients in therapeutically effective amounts to increase the sensitivity of the cells to ionizing radiation and/or to promote the treatment of diseases which are treatable with ionizing radiation.
The term “chemosensitizer” is defined as a molecule administered to patients in therapeutically effective amounts to increase the sensitivity of cells to chemotherapy and/or promote the treatment of diseases which are treatable with chemotherapeutics.
In one embodiment the compound of the invention is administered with a “radiosensitizer” and/or “chemosensitizer”. In one embodiment the compound of the invention is administered with an “immune sensitizer”.
The term “immune sensitizer” is defined as a molecule administered to patients in therapeutically effective amounts to increase the sensitivity of cells to a Polθ inhibitor.
Many cancer treatment protocols currently employ radiosensitizers in conjunction with radiation of x-rays. Examples of x-ray activated radiosensitizers include, but are not limited to, the following: metronidazole, misonidazole, desmethylmisonidazole, pimonidazole, etanidazole, nimorazole, mitomycin C, RSU 1069, SR 4233, EO9, RB 6145, nicotinamide, 5-bromodeoxyuridine (BUdR), 5-iododeoxyuridine (IUdR), bromodeoxycytidine, fluorodeoxyuridine (FudR), hydroxyurea, cisplatin, and therapeutically effective analogs and derivatives of the same.
Photodynamic therapy (PDT) of cancers employs visible light as the radiation activator of the sensitizing agent. Examples of photodynamic radiosensitizers include the following, but are not limited to: hematoporphyrin derivatives, Photofrin, benzoporphyrin derivatives, tin etioporphyrin, pheoborbide-a, bacteriochlorophyll-a, naphthalocyanines, phthalocyanines, zinc phthalocyanine, and therapeutically effective analogs and derivatives of the same.
Radiosensitizers may be administered in conjunction with a therapeutically effective amount of one or more other compounds, including but not limited to: compounds of the invention; compounds which promote the incorporation of radiosensitizers to the target cells; compounds which control the flow of therapeutics, nutrients, and/or oxygen to the target cells; chemotherapeutic agents which act on the tumour with or without additional radiation; or other therapeutically effective compounds for treating cancer or other diseases.
Chemosensitizers may be administered in conjunction with a therapeutically effective amount of one or more other compounds, including but not limited to: compounds of the invention; compounds which promote the incorporation of chemosensitizers to the target cells; compounds which control the flow of therapeutics, nutrients, and/or oxygen to the target cells; chemotherapeutic agents which act on the tumour or other therapeutically effective compounds for treating cancer or other disease. Calcium antagonists, for example verapamil, are found useful in combination with antineoplastic agents to establish chemosensitivity in tumor cells resistant to accepted chemotherapeutic agents and to potentiate the efficacy of such compounds in drug-sensitive malignancies.
Examples of immune sensitizers include the following, but are not limited to: immunomodulating agents, for example monoclonal antibodies such as immune checkpoint antibodies [e.g. CTLA-4 blocking antibodies and/or antibodies against PD-1 and PD-L1 and/or PD-L2 for example ipilimumab (CTLA4), MK-3475 (pembrolizumab, formerly lambrolizumab, anti-PD-1), nivolumab (anti-PD-1), BMS-936559 (anti-PD-L1), MPDL320A, AMP-514 or MEDI4736 (anti-PD-L1), or tremelimumab (formerly ticilimumab, CP-675,206, anti-CTLA-4)]; or Signal Transduction inhibitors; or cytokines (such as recombinant interferons); or oncolytic viruses; or immune adjuvants (e.g. BCG).
Immune sensitizers may be administered in conjunction with a therapeutically effective amount of one or more other compounds, including but not limited to: compounds of the invention; compounds which promote the incorporation of immune sensitizers to the target cells; compounds which control the flow of therapeutics, nutrients, and/or oxygen to the target cells; therapeutic agents which act on the tumour or other therapeutically effective compounds for treating cancer or other disease.
For use in combination therapy with another chemotherapeutic agent, the compound of the formula (I) and one, two, three, four or more other therapeutic agents can be, for example, formulated together in a dosage form containing two, three, four or more therapeutic agents i.e. in a unitary pharmaceutical composition containing all agents. In an alternative embodiment, the individual therapeutic agents may be formulated separately and presented together in the form of a kit, optionally with instructions for their use.
In one embodiment is provided a combination of a compound of formula (I) with one or more (e.g. 1 or 2) other therapeutic agents (e.g. anticancer agents as described above). In a further embodiment is provided a combination of a Polθ inhibitor as described herein and a PI3K/AKT pathway inhibitor selected from: apitolisib, buparlisib, Copanlisib, pictilisib, ZSTK-474, CUDC-907, GSK-2636771, LY-3023414, ipatasertib, afuresertib, MK-2206, MK-8156, Idelalisib, BEZ235 (dactolisib), BYL719, GDC-0980, GDC-0941, GDC-0032 and GDC-0068.
In another embodiment is provided a compound of formula (I) in combination with one or more (e.g. 1 or 2) other therapeutic agents (e.g. anticancer agents) for use in therapy, such as in the prophylaxis or treatment of cancer.
In one embodiment the pharmaceutical composition comprises a compound of formula (I) together with a pharmaceutically acceptable carrier and optionally one or more therapeutic agent(s).
In another embodiment the invention relates to the use of a combination according to the invention in the manufacture of a pharmaceutical composition for inhibiting the growth of tumour cells.
In a further embodiment the invention relates to a product containing a compound of formula (I) and one or more anticancer agent, as a combined preparation for simultaneous, separate or sequential use in the treatment of patients suffering from cancer.
The invention will now be illustrated, but not limited, by reference to the specific embodiments described in the following examples.
For purification of samples by HPLC the following columns were typically used; SunFire C18, Xtimate C18, Phenomenex Gemini, Phenomenex Synergi C18, Phenomenex Luna, Waters Xbridge C18, Boston Prime C18 and Shim-pack C18. Typical mobile phases used were water and MeCN, with either acidic or basic additives, such as formic acid (0.1% v/v) or ammonium hydroxide (0.05% v/v). A typical method started with 95% water:5% MeCN and decreasingly polar ratios of water and MeCN over a period of 5 to 12 min, with a typical flow rate of 25 mL/min.
Step a. To a solution of tert-butyl 2-((diphenylmethylene)amino acetate (CAS: 81477-94-3; 100 g, 339 mmol) and dimethyl fumarate (CAS: 624-49-7; 73.2 g, 508 mmol) in EtOAc (1 L) was added (S)-2-((2,3-bis(dicyclohexylamino)cycloprop-2-en-1-ylidene)amino)propan-1-ol (CAS: 1808186-23-3, prepared as described in J. S. Bander et. al. Chem. Sci. 2015, 6, 1537; 18.5 g, 33.9 mmol). The reaction mixture was stirred at rt for 48 h. Upon completion, the reaction mixture was evaporated and the crude product was purified by column chromatography (hexane/EtOAc=4/1) to give 1-(tert-butyl) 2,3-dimethyl (1S,2S)-1-((diphenylmethylene)amino)propane-1,2,3-tricarboxylate as a colourless oil (140 g, 94% yield).
Step b. To a solution of 1-(tert-butyl) 2,3-dimethyl (1S,2S)-1-((diphenylmethylene)amino)propane-1,2,3-tricarboxylate (140 g, 318.5 mmol) in THF (1.4 L) was added 15% w/w aq. citric acid (1.4 L). The reaction mixture was stirred at rt for 48 h. Upon completion, the reaction was quenched with sat. aq. NaHCO3 and extracted with EtOAc (3×1 L). The combined organic layers were dried over Na2SO4 and evaporated. The crude product was purified by column chromatography (hexane/EtOAc=3/7) to give the title compound as a white solid (70 g, 90% yield).
1H NMR (400 MHz, CDCl3) δ ppm 6.63 (br s, 1H), 4.46 (d, J=6.0 Hz, 1H), 3.77 (s, 3H), 3.37 (dt, J=6.0, 8.0, 9.2 Hz, 1H), 2.76-2.55 (m, 2H), 1.47 (s, 9H).
Step a. To a solution of 2-fluoro-6-nitroaniline (CAS: 17809-36-8; 100 g, 641 mmol) in DCM (500 mL) was added di-tert-butyl dicarbonate (308 g, 1.41 mol), TEA (130 g, 1.28 mol) and DMAP (7.83 g, 64.1 mmol). The mixture was stirred at 40° C. for 12 h. Upon completion, the reaction mixture was quenched with sat. aq. NH4Cl (500 mL) at 25° C., diluted with water (500 mL) and extracted with DCM (3×1 L). The combined organic layers were washed with brine (3×500 mL), dried over Na2SO4 and evaporated. The crude product was triturated (PE/EtOAc=20/1, 500 mL) to give tert-butyl (tert-butoxycarbonyl)(2-fluoro-6-nitrophenyl)carbamate (210 g, 92% yield) as a yellow solid.
1H NMR (400 MHz, CDCl3) δ ppm 7.87 (d, J=8.0 Hz, 1H), 7.53-7.36 (m, 2H), 1.41 (s, 18H).
Step b. To a solution of tert-butyl (tert-butoxycarbonyl)(2-fluoro-6-nitrophenyl)carbamate (210 g, 589 mmol) in DCM (2.1 L) was added TFA (87.4 g, 766 mmol) and the mixture was stirred at 25° C. for 12 h. Upon completion, the reaction mixture was quenched with sat. aq. Na2CO3 (500 mL), diluted with water (500 mL) and extracted with DCM (3×1 L). The combined organic layers were washed with brine (3×500 mL), dried over Na2SO4 and evaporated to give tert-butyl (2-fluoro-6-nitrophenyl)carbamate (145 g, 96% yield) as a yellow solid.
1H NMR (400 MHz, CDCl3) δ ppm 7.82-7.78 (m, 1H), 7.38-7.30 (m, 2H), 7.23-7.15 (m, 1H), 1.43 (s, 9H).
Step c. To a solution of tert-butyl (2-fluoro-6-nitrophenyl)carbamate (145 g, 567 mmol) in MeOH (2 L) was added 10% Pd/C (15 g) and the mixture was stirred at 25° C. for 12 h under a H2 atmosphere (15 psi). Upon completion, the reaction mixture was filtered and evaporated to give tert-butyl (2-amino-6-fluorophenyl)carbamate (128 g, 99% yield) as an off-white solid.
1H NMR (400 MHz, CDCl3) δ ppm 7.01-6.95 (m, 1H), 6.60-6.46 (m, 2H), 6.10 (br s, 1H), 4.02 (br s, 2H), 1.51 (s, 9H).
Step d. To a solution of tert-butyl (2-amino-6-fluorophenyl)carbamate (128 g, 566 mmol) in MeOH (1.5 L) was added paraformaldehyde (25.5 g, 849 mmol) and sodium methoxide (306 g, 5.66 mol). The reaction mixture was stirred at 25° C. for 12 h, and then NaBH3CN (107 g, 2.83 mol) was slowly added and stirred at 25° C. for 12 h. Upon completion, the reaction mixture was quenched with sat. aq. NH4Cl (500 mL), diluted with water (500 mL) and extracted with EtOAc (3×1 L). The combined organic layers were washed with brine (3×500 mL), dried over Na2SO4 and evaporated. The crude product was purified by column chromatography (PE/EtOAc=10/1) to give the title compound (56 g, 41% yield) as a yellow solid.
m/z ES+ [M+H]+ 241.2; 1H NMR (400 MHz, CDCl3) δ ppm 7.14-7.07 (m, 1H), 6.50-6.43 (m, 2H), 5.81 (br s, 1H), 4.49 (br s, 1H), 2.87 (s, 3H), 1.51 (s, 9H).
The title compound was prepared in a similar manner to Intermediate 2a, using 2-chloro-6-nitroaniline (CAS: 769-11-9) in step a.
1H NMR (400 MHz, CDCl3) δ ppm 7.10 (t, J=8.0 Hz, 1H), 6.76 (dd, J=1.2, 8.0 Hz, 1H), 6.58 (d, J=8.0 Hz, 1H), 5.91 (br s, 1H), 4.52 (br s, 1H), 2.87 (s, 3H), 1.51 (s, 9H).
The title compound was prepared in a similar manner to Intermediate 2a, using 4-fluoro-2-nitroaniline (CAS: 364-78-3) in step a.
1H NMR (400 MHz, CDCl3) δ ppm 7.12-7.08 (m, 1H), 6.42-6.32 (m, 2H), 5.85 (br s, 1H), 4.12 (br s, 1H), 2.83 (s, 3H), 1.50 (s, 9H).
The title compound was prepared in a similar manner to Intermediate 2a, using 2,4-difluoro-6-nitroaniline (CAS: 364-30-7) in step a.
1H NMR (400 MHz, CDCl3) δ ppm 6.24-6.07 (m, 2H), 5.66 (br s, 1H), 4.58 (br s, 1H), 2.84 (d, J=5.2 Hz, 3H), 1.49 (s, 9H).
Step a. To a solution of 4-fluoro-2-nitroaniline (16.5 g, 106 mmol) in DMF (330 mL) was added N-chlorosuccinimide (15.5 g, 116 mmol) in DMF (330 mL) and the mixture was stirred at 40° C. for 12 h. Upon completion, the reaction mixture was quenched with sat. aq. NH4Cl (100 mL) at 25° C., diluted with water (300 mL) and extracted with EtOAc (3×300 mL). The combined organic layers were washed with brine (3×200 mL), dried over Na2SO4 and evaporated. The crude product was purified by column chromatography (PE/EtOAc=50/1) to give 2-chloro-4-fluoro-6-nitroaniline (17 g, 84% yield) as a brown solid.
1H NMR (400 MHz, CDCl3) δ ppm 7.85 (dd, J=8.8, 3.2 Hz, 1H), 7.40 (dd, J=7.2, 3.2 Hz, 1H), 6.44 (br s, 2H).
Steps b-e. These 4 steps were conducted in a similar manner to Intermediate 2a, steps a-d.
m/z ES+ [M-tBu+H]+ 219.4; 1H NMR (400 MHz, CDCl3) δ ppm 6.48 (dd, J=8.4, 2.4 Hz, 1H), 6.26 (dd, J=11.2, 2.4 Hz, 1H), 5.74 (br s, 1H), 4.61 (br s, 1H), 2.84 (s, 3H) 1.50 (s, 9H).
Step a. To a solution of 2-chloro-4,5-difluoroaniline (CAS: 2613-32-3; 19.0 g, 116 mmol) in EtOAc (400 mL) was added DIPEA (30.0 g, 232 mmol) and acetic anhydride (17.8 g, 174 mmol) dropwise. The mixture was stirred at 50° C. for 16 h. Upon completion, the reaction mixture was washed with water (4×100 mL), dried over Na2SO4, and evaporated to give N-(2-chloro-4,5-difluorophenyl)acetamide (23 g, crude) as brown solid.
1H NMR (400 MHz, CDCl3) δ ppm 8.38 (dd, J=8.4, 12.4 Hz, 1H), 7.53 (br s, 1H), 7.26-7.19 (m, 1H), 2.25 (s, 3H).
Step b. To a solution of N-(2-chloro-4,5-difluorophenyl)acetamide (23.0 g, 112 mmol) in conc. H2SO4 (200 mL) was added 80% HNO3 (32.2 g, 409 mmol) at 0° C. The mixture was allowed to warm to 25° C. over 1 h. Upon completion, the reaction mixture was poured into ice water (600 mL), and extracted with EtOAc (3×200 mL). The combined organic layers were washed with brine (3×200 mL), dried over Na2SO4 and evaporated. The crude product was purified by column chromatography (PE/EtOAc=3/1) to give N-(6-chloro-3,4-difluoro-2-nitrophenyl)acetamide (26.0 g, 92% yield) as yellow solid.
1H NMR (400 MHz, CDCl3) δ ppm 7.54 (dd, J=7.6, 8.8 Hz, 1H), 7.35 (br s, 1H), 2.23 (s, 3H).
Step c. To a solution of N-(6-chloro-3,4-difluoro-2-nitrophenyl)acetamide (26.0 g, 104 mmol) in 1,4-dioxane (250 mL) was added 12 M aqueous hydrochloric acid (17 mL). The mixture was stirred at 80° C. for 18 h. Upon completion, the reaction mixture was diluted with water (60 mL) and extracted with EtOAc (2×40 mL). The combined organic layers were dried over Na2SO4, and evaporated to give 6-chloro-3,4-difluoro-2-nitroaniline (22 g, crude) as yellow solid which was used directly in the next step.
Steps d-g. These 4 steps were conducted in a similar manner to Intermediate 2a, steps a-d.
m/z ES+ [M-tBu+H]+ 236.7; 1H NMR (400 MHz, CDCl3) δ ppm 6.64-6.59 (m, 1H), 5.72 (br s, 1H), 4.31 (br s, 1H), 3.08 (d, J=4.4 Hz, 3H), 1.49 (s, 9H).
Step a. A mixture of Intermediate 1 (40.0 g, 164 mmol) and TFA (250 mL) was stirred at 25° C. for 12 h under a N2 atmosphere. Upon completion, the reaction mixture was evaporated. The residue was triturated (PE/EtOAc=10/1) for 60 min. The suspension was filtered to give (2S,3S)-3-(methoxycarbonyl)-5-oxopyrrolidine-2-carboxylic acid (27.0 g, 87% yield) as a white solid.
1H NMR (400 MHz, DMSO-d6) δ ppm 13.13 (br s, 1H), 8.15 (s, 1H), 4.26 (d, J=0.8 Hz, 1H), 3.68 (s, 3H), 3.37-3.33 (m, 1H), 2.56-2.52 (m, 1H), 2.36-2.32 (m, 1H).
Step b. To a solution of (2S,3S)-3-(methoxycarbonyl)-5-oxopyrrolidine-2-carboxylic acid (5 g, 26.7 mmol) in MeCN (50 mL) was added Ghosez's reagent (CAS: 26189-59-3; 3.93 g, 29.3 mmol) at 0° C. The mixture was stirred at rt for 1 h and then added dropwise to a mixture of Intermediate 2a (6.42 g, 26.7 mmol) and N,N-dimethylpyridin-2-amine (6.53 g, 53.4 mmol) in MeCN (50 mL) at 0° C. The reaction mixture was stirred at 0° C. for 30 min. Upon completion, the mixture was quenched with water (6 mL) and then evaporated to remove MeCN. The residue was diluted with additional water (35 mL) and extracted with EtOAc (3×50 mL). The combined organic layers were washed with brine (100 mL), dried over Na2SO4 and evaporated. The crude product was purified by column chromatography (PE/EtOAc=0/1) to give methyl (2S,3S)-2-((2-((tert-butoxycarbonyl)amino)-3-fluorophenyl)(methyl)carbamoyl)-5-oxopyrrolidine-3-carboxylate (8.7 g, 78% yield) as a white solid.
m/z ES+ [M+H]+ 410.0; 1H NMR (400 MHz, CDCl3) δ ppm 7.29 (s, 1H), 7.25-7.17 (m, 1H), 7.09 (d, J=7.6 Hz, 1H), 6.69-6.43 (m, 2H), 4.57 (d, J=4.0 Hz, 1H), 3.72-3.66 (m, 3H), 3.54-3.45 (m, 1H), 3.25-3.18 (m, 3H), 2.84 (m, 1H), 2.61-2.43 (m, 1H), 1.48 (s, 9H).
Step c. A mixture of methyl (2S,3S)-2-((2-((tert-butoxycarbonyl)amino)-3-fluorophenyl)(methyl)carbamoyl)-5-oxopyrrolidine-3-carboxylate (22.0 g, 53.7 mmol), 2-bromo-6-methyl-4-(trifluoromethyl)pyridine (CAS: 451459-17-9; 15.4 g, 64.4 mmol), Pd2(dba)3 (4.92 g, 5.37 mmol), Xantphos (6.22 g, 10.7 mmol) and K2CO3 (18.5 g, 134 mmol) in 1,4-dioxane (220 mL) was degassed and purged with N2 3 times. The reaction mixture was stirred at 100° C. for 3 h under a N2 atmosphere. Upon completion, the mixture was filtered, evaporated, water (150 mL) was added, and the mixture was extracted with EtOAc (3×170 mL). The combined organic layers were washed with brine (300 mL), dried over Na2SO4 and evaporated. The crude product was purified by column chromatography (PE/EtOAc=1/1) to give methyl (2S,3S)-2-((2-((tert-butoxycarbonyl)amino)-3-fluorophenyl)(methyl)carbamoyl)-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-5-oxopyrrolidine-3-carboxylate (27.7 g, 89% yield) as a yellow solid.
m/z ES+ [M+H]+ 569.2; 1H NMR (400 MHz, DMSO-d6) δ ppm 8.95 (s, 1H), 8.41 (s, 1H), 7.75 (d, J=8.0 Hz, 1H), 7.50-7.44 (m, 2H), 7.41-7.36 (m, 1H), 3.76 (s, 1H), 3.55 (s, 1H), 3.49 (m, 1H), 3.31 (s, 3H), 3.14-3.08 (m, 3H), 2.66 (s, 3H), 2.62-2.58 (m, 1H), 1.45 (s, 9H).
Step d. To a solution of (2S,3S)-2-((2-((tert-butoxycarbonyl)amino)-3-fluorophenyl)(methyl)carbamoyl)-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-5-oxopyrrolidine-3-carboxylate (53.4 g, 93.9 mmol) in THF (530 mL) and MeOH (53 mL) was added NaBH4 (7.11 g, 187 mmol) portion-wise at 0° C. The reaction mixture was stirred at 20° C. for 1 h. Upon completion, the mixture was slowly quenched with sat. aq. NH4Cl 50 (mL). The mixture was stirred for a further 30 min, evaporated, water (200 mL) was added and the mixture was extracted with EtOAc (3×250 mL). The combined organic layers were washed with brine (500 mL), dried over Na2SO4 and evaporated. The crude product was purified by column chromatography (PE/EtOAc=0/1) to give tert-butyl (2-fluoro-6-((2S,3S)-3-(hydroxymethyl)-N-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-5-oxopyrrolidine-2-carboxamido)phenyl)carbamate (47 g, 93% yield) as a yellow solid.
m/z ES+ [M+H]+ 541.1; 1H NMR (400 MHz, CDCl3) δ ppm 8.60 (s, 1H), 7.81 (d, J=8.0 Hz, 1H), 7.42 (m, 1H), 7.27 (s, 2H), 7.11 (s, 1H), 6.13 (s, 1H), 3.41-3.39 (m, 1H), 3.28 (s, 3H), 3.25-3.16 (m, 2H), 2.97-2.93 (m, 1H), 2.63 (s, 3H), 2.23-2.14 (m, 1H), 2.09 (d, J=17.6 Hz, 1H), 1.53 (s, 9H).
Step e. To a solution of tert-butyl (2-fluoro-6-((2S,3S)-3-(hydroxymethyl)-N-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-5-oxopyrrolidine-2-carboxamido)phenyl)carbamate (47.0 g, 86.9 mmol) in DCM (500 mL) was added TEA (35.2 g, 347 mmol) at 0° C. Then MsCl (14.9 g, 130 mmol) was added dropwise at 0° C. The reaction mixture was stirred at 0° C. for 1 h. Upon completion, the reaction mixture was quenched with water (400 mL) and extracted with EtOAc (2×400 mL). The combined organic layers were washed with brine (800 mL), dried over Na2SO4 and evaporated to give ((2S,3S)-2-((2-((tert-butoxycarbonyl)amino)-3-fluorophenyl)(methyl)carbamoyl)-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-5-oxopyrrolidin-3-yl)methyl methanesulfonate (52.0 g, 98% yield) as a yellow solid.
m/z ES+ [M+H]+ 619.1.
Step f. To a solution of ((2S,3S)-2-((2-((tert-butoxycarbonyl)amino)-3-fluorophenyl)(methyl)carbamoyl)-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-5-oxopyrrolidin-3-yl)methyl methanesulfonate (26.0 g, 42.0 mmol) in NMP (400 mL) was added K3PO4 (26.7 g, 126 mmol). The mixture was stirred at 60° C. for 12 h. Upon completion, the mixture was diluted with water (1.2 L) and extracted with EtOAc (3×800 mL). The combined organic layers were washed with water (2×1 L), brine (2×1 L), dried over Na2SO4 and evaporated. The crude product was purified by column chromatography (PE/EtOAc=3/1) to give tert-butyl (3aR,11aS)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-5-carboxylate (31.0 g, 67% yield) as a yellow solid.
m/z ES+ [M+H]+ 523.0.
Step g. To a solution of tert-butyl (3aR,11aS)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-5-carboxylate (31.0 g, 59.3 mmol) in DCM (320 mL) was added TFA (123 g, 1.08 mol). The mixture was stirred at 20° C. for 2 h. Upon completion, the mixture was basified to pH 8 with sat. aq. NaHCO3. The organic layer was separated and the aqueous was further extracted with EtOAc (3×600 mL). The combined organic layers were washed with brine (2×500 mL), dried over Na2SO4 and evaporated to give the title compound (22.3 g, 90% yield) as a yellow solid.
m/z ES+ [M+H]+ 423.1; 1H NMR (400 MHz, CDCl3) δ ppm 8.35 (s, 1H), 7.32-7.27 (m, 1H), 7.24-7.20 (m, 1H), 7.20-7.14 (m, 1H), 7.06 (s, 1H), 4.79 (d, J=9.2 Hz, 1H), 3.78-3.73 (m, 1H), 3.36 (s, 3H), 2.87-2.70 (m, 3H), 2.50 (s, 3H), 2.29-2.21 (m, 1H).
The title compound was prepared in a similar manner to Intermediate 5a, using Intermediate 2b in step b.
1H NMR (400 MHz, CDCl3) δ ppm 8.36 (s, 1H), 7.50 (dd, J=1.6, 8.0 Hz, 1H), 7.39-7.33 (m, 1H), 7.30-7.25 (m, 1H), 7.08 (s, 1H), 4.78 (d, J=9.2 Hz, 1H), 3.75 (dd, J=4.0, 13.2 Hz, 1H), 3.37 (s, 3H), 2.93-2.74 (m, 3H), 2.52 (s, 3H), 2.33-2.23 (m, 1H).
The title compound was prepared in a similar manner to Intermediate 5a, using Intermediate 2c in step b.
1H NMR (400 MHz, CDCl3) δ ppm 8.35 (s, 1H), 7.28 (d, J=6.0 Hz, 1H), 7.15-7.03 (m, 3H), 4.83-4.74 (m, 1H), 3.79 (d, J=8.8 Hz, 1H), 3.36 (s, 3H), 2.87-2.68 (m, 3H), 2.52 (s, 3H), 2.27-2.20 (m, 2H).
The title compound was prepared in a similar manner to Intermediate 5a, using Intermediate 2d in step b.
m/z ES+ [M+H]+ 441.1.
The title compound was prepared in a similar manner to Intermediate 5a, using Intermediate 3 in step b.
m/z ES+ [M+H]+ 457.1; 1H NMR (400 MHz, CDCl3) δ ppm 8.35 (s, 1H), 7.25 (d, J=2.8 Hz, 1H), 7.10-7.04 (m, 2H), 4.73 (d, J=9.2 Hz, 1H), 3.72 (dd, J=13.2, 3.6 Hz, 1H), 3.34 (s, 4H), 2.89-2.75 (m, 2H), 2.74-2.67 (m, 1H), 2.51 (s, 3H), 2.32-2.20 (m, 1H).
The title compound was prepared in a similar manner to Intermediate 5a, using Intermediate 4 in step b.
m/z ES+ [M+H]+ 475.0.
Step a. To a solution of Intermediate 5e (200 mg, 0.44 mmol), methylboronic acid (262 mg, 4.38 mmol) and Na2CO3 (139 mg, 1.31 mmol) in 1,4-dioxane (1.5 mL) and water (0.2 mL) was added XPhos-Pd-G2 (34.4 mg, 0.044 mmol). The mixture was degassed and purged with N2 3 times. The reaction mixture was stirred at 110° C. for 1 h under a N2 atmosphere. Upon completion, the reaction mixture was diluted with water (5 mL) and extracted with EtOAc (3×25 mL). The combined organic layers were washed with brine (5 mL), dried over Na2SO4, and evaporated. The residue was purified by Prep-TLC (PE/EtOAc=3/1) to give the title compound (150 mg, 72% yield) as yellow solid.
1H NMR (400 MHz, CDCl3) δ ppm 8.35 (s, 1H), 7.07 (s, 1H), 6.97 (m, 2H), 4.83 (d, J=9.2 Hz, 1H), 3.73 (dd, J=4.8, 13.6 Hz, 1H), 3.34 (s, 3H), 2.84-2.71 (m, 3H), 2.62-2.54 (m, 1H), 2.51 (s, 3H), 2.42 (s, 3H), 2.27-2.16 (m, 1H).
The title compound was prepared in a similar manner to Intermediate 6a, using cyclopropylboronic acid in step a.
m/z ES+ [M+H]+ 463.2; 1H NMR (400 MHz, CDCl3) δ ppm 8.35 (s, 1H), 7.07 (s, 1H), 6.91 (dd, J=2.8, 8.8 Hz, 1H), 6.59 (dd, J=2.8, 9.6 Hz, 1H), 4.84 (d, J=8.4 Hz, 1H), 3.77 (m, 1H), 3.34 (s, 3H), 3.09-2.90 (m, 1H), 2.86-2.69 (m, 3H), 2.51 (s, 3H), 2.33-2.18 (m, 2H), 1.18-1.05 (m, 2H), 0.80-0.63 (m, 2H).
The title compound was prepared in a similar manner to Intermediate 6a, using Intermediate 5b and methylboronic acid in step a.
m/z ES+ [M+H]+ 418.9; 1H NMR (400 MHz, CDCl3) δ ppm 8.34 (s, 1H), 7.26-7.14 (m, 3H), 7.05 (s, 1H), 4.85 (d, J=9.2 Hz, 1H), 3.81-3.69 (m, 1H), 3.37 (s, 3H), 2.84-2.72 (m, 2H), 2.67-2.58 (m, 1H), 2.51 (s, 3H), 2.42 (s, 3H), 2.28-2.16 (m, 1H).
Step a. A mixture of Intermediate 5a (7.6 g, 17.9 mmol), Na2CO3 (5.72 g, 53.9 mmol), TBAB (580 mg, 1.80 mmol) in DMF (70 mL) was degassed and purged with N2 3 times, and then 3-bromoprop-1-ene (8.71 g, 71.9 mmol) was added dropwise at 20° C. The mixture was stirred at 100° C. for 2 h under a N2 atmosphere. Upon completion, the reaction mixture was diluted with water (200 mL) and extracted with EtOAc (3×200 mL). The combined organic layers were washed with water (3×200 mL), brine (2×200 mL), dried over Na2SO4 and evaporated. The crude product was purified by column chromatography (PE/EtOAc=5/1) to give (3aR,11aS)-5-allyl-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (7 g, 82% yield) as a yellow solid.
m/z ES+ [M+H]+ 463.2; 1H NMR (400 MHz, CDCl3) δ ppm 8.32 (s, 1H), 7.36-7.29 (m, 1H), 7.23-7.21 (m, 1H), 7.15-7.11 (m, 1H), 7.05 (s, 1H), 5.78-5.65 (m, 1H), 5.23-5.18 (m, 1H), 5.06 (dd, J=1.6, 10.0 Hz, 1H), 4.66-4.61 (m, 1H), 3.77-3.63 (m, 2H), 3.59-3.49 (m, 1H), 3.34 (s, 3H), 2.92-2.82 (m, 2H), 2.75-2.66 (m, 1H), 2.50 (s, 3H), 2.24-2.14 (m, 1H).
Step b. To a solution of (3aR,11aS)-5-allyl-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (7.0 g, 15.1 mmol) in THF (140 mL) and water (70 mL) was added NaIO4 (9.71 g, 45.4 mmol) and OsO4 (385 mg, 1.51 mmol) at 0° C. The mixture was stirred at 0° C. for 1 h. Upon completion, sat. aq. Na2S2O3 (100 mL) was added, stirred at rt for a further 30 min and then the aqueous mixture was extracted with EtOAc (3×200 mL). The combined organic layers were washed with sat. aq. Na2S2O3 (2×150 mL), sat. NaHCO3 (2×150 mL), brine (300 mL), dried over Na2SO4 and evaporated to give the title compound (7 g, crude) as a yellow solid.
m/z ES+ [M+H]+ 465.2.
The title compound was prepared in a similar manner to Intermediate 7a, using Intermediate 5b in step a.
1H NMR (400 MHz, CDCl3) δ ppm 9.67 (t, J=1.6 Hz, 1H), 8.34 (s, 1H), 7.52-7.46 (m, 1H), 7.39-7.31 (m, 2H), 7.06 (s, 1H), 4.68 (d, J=9.2 Hz, 1H), 3.88-3.71 (m, 2H), 3.56 (dd, J=4.8, 14.8 Hz, 1H), 3.37 (s, 3H), 3.15 (dd, J=12.0, 14.4 Hz, 1H), 3.07-2.94 (m, 1H), 2.75 (dd, J=8.8, 16.8 Hz, 1H), 2.50-2.46 (m, 3H), 2.24 (dd, J=11.6, 16.8 Hz, 1H).
The title compound was prepared in a similar manner to Intermediate 7a, using Intermediate 5c in step a.
m/z ES+ [M+H]+ 465.2.
The title compound was prepared in a similar manner to Intermediate 7a, using Intermediate 5d in step a.
m/z ES+ [M+H]+ 483.4.
The title compound was prepared in a similar manner to Intermediate 7a, using Intermediate 5e in step a.
m/z ES+ [M+H]+ 499.2.
The title compound was prepared in a similar manner to Intermediate 7a, using Intermediate 5f in step a.
m/z ES+ [M+H]+ 516.9.
The title compound was prepared in a similar manner to Intermediate 7a, using Intermediate 6a in step a.
1H NMR (400 MHz, CDCl3) δ ppm 9.63 (s, 1H), 8.35 (s, 1H), 7.07 (s, 1H), 7.02-6.98 (m, 1H), 6.93 (dd, J=2.8, 8.4 Hz, 1H), 4.75 (d, J=9.2 Hz, 1H), 3.84 (d, J=18.4 Hz, 1H), 3.67-3.57 (m, 2H), 3.36 (s, 3H), 3.01-2.85 (m, 2H), 2.75 (dd, J=8.4, 16.8 Hz, 1H), 2.50 (s, 6H), 2.25-2.18 (m, 1H).
The title compound was prepared in a similar manner to Intermediate 7a, using Intermediate 6b in step a.
m/z ES+ [M+H]+ 505.4.
The title compound was prepared in a similar manner to Intermediate 7a, using Intermediate 6c in step a.
m/z ES+ [M+H]+ 461.3; 1H NMR (400 MHz, CDCl3) δ ppm 9.65 (t, J=1.6 Hz, 1H), 8.34 (s, 1H), 7.27-7.19 (m, 3H), 7.05 (s, 1H), 4.75 (d, J=9.2 Hz, 1H), 3.84 (dd, J=18.0, 1.6 Hz, 1H), 3.68-3.59 (m, 2H), 3.40-3.35 (m, 3H), 3.04-2.89 (m, 2H), 2.78-2.71 (m, 1H), 2.50 (d, J=2.4 Hz, 6H), 2.26-2.18 (m, 1H).
Step a. To a solution of Intermediate 7a (1.00 g, 2.15 mmol) in MeOH (10 mL) was added NaBH3CN (244 mg, 6.46 mmol) and the mixture was stirred at 0° C. for 30 min. Upon completion, the reaction mixture was quenched with sat. aq. NH4Cl (20 mL) at 0° C. The mixture was diluted with water (30 mL) and extracted with EtOAc (3×50 mL). The combined organic layers were washed with brine (3×20 mL), dried over Na2SO4 and evaporated to give (3aR,11aS)-6-fluoro-5-(2-hydroxyethyl)-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (1 g, 99% yield) as a yellow solid.
m/z ES+ [M+H]+ 467.2; 1H NMR (400 MHz, CDCl3) δ ppm 8.33 (s, 1H), 7.41-7.33 (m, 1H), 7.24 (d, J=8.0 Hz, 1H), 7.17 (t, J=9.2 Hz, 1H), 7.06 (s, 1H), 4.65 (d, J=9.2 Hz, 1H), 4.13 (q, J=7.2 Hz, 1H), 3.59 (dd, J=13.6, 4.4 Hz, 1H), 3.55-3.42 (m, 2H), 3.39-3.35 (m, 3H), 3.32-3.23 (m, 2H), 3.05-2.95 (m, 1H), 2.94-2.80 (m, 1H), 2.75 (dd, J=16.8, 8.2 Hz, 1H), 2.50 (s, 3H), 2.27-2.17 (m, 1H).
Step b. To a solution of (3aR,11aS)-6-fluoro-5-(2-hydroxyethyl)-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (1 g, 2.14 mmol) in THF (10 mL) was added I2 (816 mg, 3.22 mmol), PPh3 (844 mg, 3.22 mmol) and imidazole (292 mg, 4.29 mmol). The reaction mixture was stirred at 0° C. for 12 h. Upon completion, the reaction mixture was quenched with sat. aq. NH4Cl (20 mL), diluted with water (30 mL) and extracted with EtOAc (3×50 mL). The combined organic layers were washed with brine (3×30 mL), dried over Na2SO4 and evaporated. The crude product was purified by column chromatography (PE/EtOAc=4/1) to give the title compound (1 g, 80% yield) as a yellow solid.
m/z ES+ [M+H]+ 577.1.
Step a. To a solution of Intermediate 5a (300 mg, 0.71 mmol) and tert-butyl 2-bromoacetate (277 mg, 1.42 mmol) in NMP (4 mL) was added K2CO3 (294 mg, 2.13 mmol) and TBAl (26.2 mg, 0.071 mmol). The mixture was stirred at 120° C. for 24 h. Upon completion, the reaction mixture was diluted with EtOAc (50 mL) and washed with brine (3×10 mL). The organic layer was dried over Na2SO4 and evaporated. The crude product was purified by column chromatography (PE/EtOAc=5/1) to give tert-butyl 2-((3aR,11aS)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)acetate (210 mg, 54% yield) as a yellow gum.
1H NMR (400 MHz, CDCl3) δ ppm 8.33 (s, 1H), 7.38-7.33 (m, 1H), 7.22 (d, J=8.0 Hz, 1H), 7.17-7.14 (m, 1H), 7.05 (s, 1H), 4.65 (d, J=9.2 Hz, 1H), 3.87-3.76 (m, 1H), 3.68-3.56 (m, 2H), 3.38-3.34 (m, 3H), 3.03-2.90 (m, 2H), 2.85 (s, 1H), 2.77-2.70 (m, 1H), 2.49 (s, 3H), 1.43 (s, 9H).
Step b. To a solution of tert-butyl 2-((3aR,11aS)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)acetate (100 mg, 0.19 mmol) in DCM (2 mL) was added TFA (3.08 g, 27.0 mmol). The mixture was stirred at 20° C. for 30 min. Upon completion, the reaction mixture was evaporated to give the title compound (100 mg, crude) as a yellow oil.
m/z ES+ [M+H]+ 481.3.
The title compound was prepared in a similar manner to Intermediate 9a, using Intermediate 5b in step a.
m/z ES+ [M+H]+ 497.1.
Step a. To a solution of Intermediate 7a (100 mg, 0.22 mmol), Intermediate A3 (99 mg, 0.32 mmol) in MeOH (2 mL), 4 Å molecular sieves (100 mg) and acetic acid (65 mg, 1.08 mmol) were added. The mixture was stirred at 25° C. for 30 min, after which, NaBH3CN (27 mg, 0.43 mmol) was added. The mixture was stirred at 25° C. for 30 min. Upon completion, the mixture was diluted with DCM (5 mL), and basified to pH 8 with sat. aq. NaHCO3. The organic phase was separated and the aqueous phase was further extracted with DCM (2×5 mL). The combined organic layers were dried over Na2SO4 and evaporated. The residue was purified by column chromatography (PE/EtOAc=1/4) and further purified by chiral SFC (column: Daicel ChiralPak AS (250×30 mm, 10 μm); mobile phase: A: 0.1% NH4OH/MeOH; B %: 50% CO2) to give two products.
Intermediate 10a: benzyl 3-((R)-4-(2-((3aR,11aS)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)ethyl)-2-(fluoromethyl)piperazin-1-yl)azetidine-1-carboxylate (22 mg, 22% yield) as a yellow solid.
m/z ES+ [M+H]+ 756.3; 1H NMR (400 MHz, CDCl3) δ ppm 8.33 (s, 1H), 7.37-7.29 (m, 6H), 7.22 (d, J=8.0 Hz, 1H), 7.17-7.11 (m, 1H), 7.06 (s, 1H), 5.09 (s, 2H), 4.71-4.64 (m, 0.5H), 4.60 (d, J=9.2 Hz, 1H), 4.58-4.53 (m, 0.5H), 4.42-4.38 (m, 0.5H), 4.30-4.28 (m, 0.5H), 4.05-3.98 (m, 2H), 3.96-3.85 (m, 2H), 3.68-3.59 (m, 1H), 3.58-3.53 (m, 1H), 3.35 (s, 3H), 3.22-3.12 (m, 2H), 2.96-2.78 (m, 3H), 2.76-2.64 (m, 2H), 2.52-2.42 (m, 6H), 2.42-2.32 (m, 3H), 2.31-2.24 (m, 1H), 2.24-2.15 (m, 1H).
Intermediate 10b: Benzyl 3-((S)-6-fluoro-4-(2-((3aR,11aS)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)ethyl)-1,4-diazepan-1-yl)azetidine-1-carboxylate (50 mg, 48% yield) as a yellow solid.
m/z ES+ [M+H]+ 756.3; 1H NMR (400 MHz, CDCl3) δ ppm 8.32 (s, 1H), 7.39-7.29 (m, 6H), 7.22 (d, J=8.0 Hz, 1H), 7.14 (dt, J=1.2, 9.2 Hz, 1H), 7.05 (s, 1H), 5.09 (s, 2H), 4.65 (t, J=5.2 Hz, 0.5H), 4.62-4.57 (m, 1H), 4.52 (t, J=5.2 Hz, 0.5H), 4.05-3.96 (m, 2H), 3.78 (m, 2H), 3.62-3.53 (m, 1H), 3.40 (t, J=5.2 Hz, 1H), 3.35 (s, 3H), 3.17 (t, J=6.8 Hz, 2H), 2.96-2.84 (m, 4H), 2.79-2.67 (m, 4H), 2.66-2.58 (m, 1H), 2.58-2.50 (m, 3H), 2.49 (s, 3H), 2.45-2.36 (m, 1H), 2.23-2.11 (m, 1H).
Step a. To a solution of Intermediate 5a (400 mg, 0.95 mmol) and 2-bromoacetonitrile (1.14 g, 9.47 mmol) in DMF (5 mL) was added Na2CO3 (402 mg, 3.79 mmol) and TBAB (31 mg, 0.095 mmol). The reaction mixture was stirred at 100° C. for 12 h. Upon completion, the reaction mixture was diluted with water (20 mL) and extracted with EtOAc (3×30 mL). The combined organic layers were washed with brine (20 mL), dried over Na2SO4, evaporated and purified by column chromatography (PE/EtOAc=2/1) to give 2-((3aR,11aS)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)acetonitrile (410 mg, 94% yield) as a brown solid.
m/z ES+ [M+H]+ 462.3; 1H NMR (400 MHz, CDCl3) δ ppm 8.33 (s, 1H), 7.47-7.44 (m, 1H), 7.31 (d, J=8.0 Hz, 1H), 7.24-7.22 (m, 1H), 7.07 (s, 1H), 4.61 (d, J=8.8 Hz, 1H), 4.21-4.14 (m, 1H), 4.04-3.96 (m, 1H), 3.62-3.58 (m, 1H), 3.42 (s, 3H), 3.09-3.00 (m, 1H), 2.97-2.92 (m, 1H), 2.79-2.73 (m, 1H), 2.50 (s, 3H), 2.22 (dd, J=16.8, 11.6 Hz, 1H).
Step b. To a solution of 2-((3aR,11aS)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)acetonitrile (200 mg, 0.43 mmol) in MeOH (4 mL) was added sodium methoxide (30 mg, 0.56 mmol) and tert-butyl 3-(hydrazinecarbonyl)azetidine-1-carboxylate (CAS: 1001907-44-3; 280 mg, 1.3 mmol), the mixture was stirred at 80° C. for 12 h. Upon completion, the reaction mixture was quenched with sat. aq. NH4Cl (20 mL), diluted with water (30 mL) and extracted with EtOAc (3×50 mL). The combined organic layers were washed with brine (3×20 mL), dried over Na2SO4, evaporated and purified by column chromatography (EtOAc) to give tert-butyl 3-(3-(((3aR,11aS)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)methyl)-1H-1,2,4-triazol-5-yl)azetidine-1-carboxylate (110 mg, 39% yield) as a yellow solid.
m/z ES+ [M+H]+ 659.5; 1H NMR (400 MHz, CDCl3) δ ppm 8.32 (s, 1H), 7.40-7.36 (m, 1H), 7.26-7.17 (m, 2H), 7.07 (s, 1H), 4.67 (d, J=8.8 Hz, 1H), 4.55-4.47 (m, 1H), 4.42-4.35 (m, 1H), 4.29-4.21 (m, 2H), 4.11-4.02 (m, 2H), 3.89-3.76 (m, 1H), 3.64-3.61 (m, 1H), 3.33 (s, 3H), 3.10-3.02 (m, 1H), 3.00-2.98 (m, 1H), 2.73 (dd, J=16.8, 8.4 Hz, 1H), 2.51 (s, 3H), 2.21 (dd, J=16.8, 11.6 Hz, 1H), 1.44 (s, 9H).
Step c. To a solution of tert-butyl 3-(3-(((3aR,11aS)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)methyl)-1H-1,2,4-triazol-5-yl)azetidine-1-carboxylate (100 mg, 0.15 mmol) in DMF (3 mL) was added K2CO3 (63 mg, 0.46 mmol) and MeI (43 mg, 0.30 mmol). The reaction mixture was stirred at 25° C. for 2 h. Upon completion, the reaction mixture was diluted with water (20 mL) and extracted with EtOAc (3×30 mL). The combined organic layers were washed with brine (20 mL), dried over Na2SO4, evaporated and purified by prep-TLC (EtOAc/MeOH=10/1) to give two products. The regiochemistry was confirmed by 2D NMR.
Intermediate 11a: tert-butyl 3-(5-(((3aR,11aS)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)methyl)-1-methyl-1H-1,2,4-triazol-3-yl)azetidine-1-carboxylate (50 mg, 49% yield) as a yellow solid.
m/z ES+ [M+H]+ 673.2; 1H NMR (400 MHz, CDCl3) δ ppm 8.30 (s, 1H), 7.39-7.36 (m, 1H), 7.22-7.15 (m, 2H), 7.10 (s, 1H), 4.61 (d, J=8.8 Hz, 1H), 4.40 (d, J=8.8 Hz, 1H), 4.36-4.33 (m, 1H), 4.23-4.49 (m, 2H), 4.19-4.07 (m, 2H), 3.84 (s, 3H), 3.77-3.74 (m, 1H), 3.59-3.27 (m, 1H), 2.99 (s, 3H), 2.99-2.96 (m, 1H), 2.99-2.96 (m, 1H), 2.95-2.73 (m, 1H), 2.72 (s, 3H), 2.20-2.19 (m, 1H), 1.47 (s, 9H).
Intermediate 11b: tert-butyl 3-(3-(((3aR,11aS)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)methyl)-1-methyl-1H-1,2,4-triazol-5-yl)azetidine-1-carboxylate (20 mg, 20% yield) as a yellow solid.
m/z ES+ [M+H]+ 673.2; 1H NMR (400 MHz, CDCl3) δ ppm 8.30 (s, 1H), 7.34-7.28 (m, 1H), 7.20-7.10 (m, 2H), 7.04 (s, 1H), 4.64 (d, J=9.2 Hz, 1H), 4.37-4.28 (m, 2H), 4.23-4.13 (m, 2H), 4.10-4.03 (m, 2H), 3.86-3.73 (m, 3H), 3.70 (s, 3H), 3.30 (s, 3H), 3.03-2.98 (m, 1H), 2.69 (dd, J=16.8, 8.4 Hz, 1H), 2.48 (s, 3H), 2.19 (dd, J=16.8, 11.6 Hz, 1H), 1.43 (s, 9H).
Step a. This step was conducted in a similar manner to Intermediate 7a, step a, using Intermediate 5b.
Step b. To a mixture of (3aR,11aS)-5-allyl-6-chloro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (100 mg, 0.21 mmol) in DCM (5 mL) was added m-CPBA (144 mg, 0.63 mmol). The reaction mixture was stirred at 30° C. for 20 h. Upon completion, the reaction mixture was diluted with water (50 mL) and extracted with EtOAc (3×30 mL). The organic layers were combined, washed with brine (3×50 mL), dried over Na2SO4 and evaporated. Purification by Prep-TLC (PE/EtOAc=1/2) gave (3aR,11aS)-6-chloro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-5-(oxiran-2-ylmethyl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (90 mg, 87% yield) as a yellow solid.
m/z ES+ [M+H]+ 495.0; 1H NMR (400 MHz, CDCl3) δ ppm 8.34 (s, 1H), 7.48-7.45 (m, 1H), 7.36-7.30 (m, 2H), 7.05 (s, 1H), 4.67-4.62 (m, 1H), 3.72-3.62 (m, 1H), 3.39 (d, J=2.4 Hz, 3H), 3.36-3.30 (m, 1H), 3.13-2.96 (m, 4H), 2.80-2.68 (m, 2H), 2.53-2.50 (m, 1H), 2.49 (s, 3H), 2.27-2.18 (m, 1H).
Step c. To a mixture of (3aR,11aS)-6-chloro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-5-(oxiran-2-ylmethyl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (208 mg, 0.42 mmol) in MeOH (5 mL) was added tert-butyl 3-(methylamino)azetidine-1-carboxylate (CAS 454703-20-9; 391 mg, 2.10 mmol). The reaction mixture was stirred at 70° C. for 18 h. Upon completion, the reaction mixture was evaporated and the residue was purified by Prep-HPLC to give tert-butyl 3-(((rac)-3-((3aR,11aS)-6-chloro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)-2-hydroxypropyl)(methyl)amino)azetidine-1-carboxylate (150 mg, 52% yield) as a yellow solid.
m/z ES+ [M+H]+ 681.4; 1H NMR (400 MHz, CDCl3) δ ppm 8.34 (s, 1H), 7.49-7.44 (m, 1H), 7.38-7.28 (m, 2H), 7.06 (s, 1H), 4.68-4.58 (m, 1H), 4.00-3.89 (m, 2H), 3.87-3.71 (m, 3H), 3.68-3.60 (m, 1H), 3.46-3.26 (m, 5H), 3.24-2.86 (m, 5H), 2.81-2.70 (m, 1H), 2.48 (s, 3H), 2.45-2.35 (m, 1H), 2.34-2.17 (m, 4H), 1.44 (s, 9H).
The racemic material (150 mg) was purified by chiral SFC (column: Daicel ChiralPak IC (250×30 mm, 10 μm); mobile phase: A: 0.1% NH4OH/MeOH; B: 55% CO2) to give two products.
Intermediate 12a: tert-butyl 3-(((S/R)-3-((3aR,11aS)-6-chloro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)-2-hydroxypropyl)(methyl)amino)azetidine-1-carboxylate (60 mg, 40% yield) as a colourless oil.
m/z ES+ [M+H]+ 681.3.
Intermediate 12b: tert-butyl 3-(((R/S)-3-((3aR,11aS)-6-chloro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)-2-hydroxypropyl)(methyl)amino)azetidine-1-carboxylate (75 mg, 50% yield) as a colourless oil.
m/z ES+ [M+H]+ 681.3.
Step a. This step was conducted as described in Intermediate 7a, step a.
Step b. To a solution of (3aR,11aS)-5-allyl-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (100 mg, 0.22 mmol) in tetrahydrofuran (3 mL) was added a solution of borane dimethyl sulfide complex (10 M in THF, 65 μL) in DCM (0.59 mL). The reaction mixture was stirred at 25° C. for 2 h. The reaction mixture was quenched with methanol (2 mL) and evaporated. The residue was dissolved in THF (4 mL) and 1 M sodium hydroxide (0.65 mL) and 30% hydrogen peroxide (221 mg, 1.95 mmol) were added. The resulting mixture was stirred at 25° C. for 1 hr. Upon completion, the reaction mixture was diluted with water (30 mL) and extracted with EtOAc (30 mL). The organic layer was washed with water (2×30 mL), 1 M sodium thiosulfate (30 mL), brine (30 mL), dried over Na2SO4 and evaporated. The residue was purified by Prep-HPLC to give (3aR,11aS)-6-fluoro-5-(3-hydroxypropyl)-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (14 mg, 69% yield) as a white solid.
m/z ES+ [M+H]+ 481.1; 1H NMR (400 MHz, CDCl3) δ ppm 8.32 (s, 1H), 7.35-7.31 (m, 1H), 7.22 (d, J=8.0 Hz, 1H), 7.18-7.14 (m, 1H), 7.05 (s, 1H), 4.63 (d, J=9.0 Hz, 1H), 3.71 (t, J=6.0 Hz, 2H), 3.62-3.54 (m, 1H), 3.37 (s, 3H), 3.29-3.17 (m, 2H), 2.98-2.85 (m, 2H), 2.79-2.67 (m, 1H), 2.49 (s, 3H), 2.27-2.14 (m, 1H), 1.67-1.55 (m, 2H).
Step c. To a solution of (3aR,11aS)-6-fluoro-5-(3-hydroxypropyl)-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (300 mg, 0.62 mmol) in DCM (5 mL) was added DMP (397 mg, 0.94 mmol). The reaction mixture was stirred at 25° C. for 1 h. Upon completion, the reaction mixture was quenched with sat. aq. Na2S2O3 (20 mL), diluted with water (30 mL) and extracted with DCM (3×50 mL). The combined organic layers were washed with sat. aq. NaHCO3 (20 mL), dried over Na2SO4 and evaporated to give 3-((3aR,11aS)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)propanal (290 mg, crude) as a yellow solid.
m/z ES+ [M+H]+ 479.2.
Step d. To a solution of 3-((3aR,11aS)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)propanal (0.29 g, 0.61 mmol) and tert-butyl 3-aminoazetidine-1-carboxylate (CAS: 193269-78-2; 209 mg, 1.21 mmol) in MeOH (5 mL) was added acetic acid (182 mg, 3.03 mmol), 4 Å molecular sieves (300 mg) and NaBH(OAc)3 (257 mg, 1.21 mmol). The reaction mixture was stirred at 25° C. for 12 h. Upon completion, the reaction mixture was evaporated and purified by column chromatography (EtOAc) to give the title compound (370 mg, 96% yield) as a yellow solid.
m/z ES+ [M+H]+ 635.2; 1H NMR (400 MHz, CDCl3) δ ppm 8.32 (s, 1H), 7.37-7.29 (m, 1H), 7.21 (d, J=8.0 Hz, 1H), 7.14 (t, J=9.2 Hz, 1H), 7.06 (s, 1H), 4.61 (d, J=8.8 Hz, 1H), 4.10-4.03 (m, 2H), 3.83-3.74 (m, 2H), 3.60 (dd, J=9.2, 5.2 Hz, 4H), 3.55 (d, J=8.8 Hz, 1H), 3.35 (s, 3H), 3.21-3.09 (m, 2H), 2.95-2.84 (m, 2H), 2.75-2.62 (m, 1H), 2.65-2.54 (m, 2H), 2.49 (s, 3H), 2.25-2.19 (m, 1H), 1.43 (s, 9H).
Step a. A mixture of Intermediate 1 (100 g, 411 mmol), 2-bromo-6-methyl-4-(trifluoromethyl)pyridine (CAS: 451459-17-9; 128 g, 534 mmol), K2CO3 (114 g, 822 mmol), CuI (15.7 g, 82.2 mmol) and N1,N2-dimethylethane-1,2-diamine (14.5 g, 164.4 mmol) in toluene (1.4 L) was stirred at 110° C. for 12 h under a N2 atmosphere. Upon completion, the reaction mixture was filtered. The filter cake was washed with EtOAc (2 L). The combined filtrate was washed with water (2 L), brine (2 L), dried over Na2SO4, evaporated and purified by column chromatography (PE/EtOAc=8/1) to give 2-(tert-butyl) 3-methyl (2S,3S)-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-5-oxopyrrolidine-2,3-dicarboxylate (108 g, 63% yield) as a yellow solid.
m/z ES+ [M+H]+ 403.2; 1H NMR (400 MHz, CDCl3) δ ppm 8.46 (s, 1H), 7.11 (s, 1H), 5.21 (d, J=2.8 Hz, 1H), 3.80 (s, 3H), 3.25-3.16 (m, 1H), 3.10-2.93 (m, 2H), 2.50 (s, 3H), 1.46 (s, 9H).
Step b. Note: The following process was conducted in 6 parallel batches.
To a solution of 2-(tert-butyl) 3-methyl (2S,3S)-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-5-oxopyrrolidine-2,3-dicarboxylate (50 g, 124 mmol) in THF (450 mL) and MeOH (50 mL) was added NaBH4 (7.05 g, 186 mmol) portion-wise at 0° C. The mixture was stirred at 0-5° C. for 3 h. Upon completion, the reaction mixture was quenched with sat. aq. NH4Cl (3 L) and extracted with EtOAc (2×3 L). The organic layers from the parallel batches were combined, washed with brine (3 L), dried over Na2SO4 and evaporated to give tert-butyl (2S,3S)-3-(hydroxymethyl)-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-5-oxopyrrolidine-2-carboxylate (300 g, crude) as a yellow gum.
m/z ES+ [M+H]+ 375.0; 1H NMR (400 MHz, CDCl3) δ ppm 8.49 (s, 1H), 7.09 (s, 1H), 4.82 (d, J=3.2 Hz, 1H), 3.77 (d, J=6.0 Hz, 2H), 2.99-2.87 (m, 1H), 2.58-2.51 (m, 2H), 2.49 (s, 3H), 2.24-2.08 (m, 1H), 1.43 (s, 9H).
Step c. To a solution of tert-butyl (2S,3S)-3-(hydroxymethyl)-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-5-oxopyrrolidine-2-carboxylate (100 g, 267 mmol) and TEA (54.1 g, 534 mmol) in DCM (1 L) was added MsCl (33.7 g, 294 mmol) dropwise at 0° C. The reaction mixture was stirred at 20° C. for 1 h. Upon completion, the reaction mixture was diluted with water (1 L). The layers were separated, the organic layer was washed with brine (3×1 L), dried over Na2SO4, filtered through a pad of silica gel (100-200 mesh) and evaporated to give tert-butyl (2S,3S)-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-3-(((methylsulfonyl)oxy)methyl)-5-oxopyrrolidine-2-carboxylate (120 g, 99% yield) as a yellow gum.
m/z ES+ [M+H]+ 453.0; 1H NMR (400 MHz, CDCl3) δ ppm 8.47 (s, 1H), 7.12 (s, 1H), 4.81 (d, J=3.2 Hz, 1H), 4.33 (d, J=6.0 Hz, 2H), 3.08 (s, 3H), 3.06-2.96 (m, 1H), 2.84-2.75 (m, 1H), 2.60-2.53 (m, 1H), 2.51 (s, 3H), 1.45 (s, 9H).
Step d. A mixture of tert-butyl (2S,3S)-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-3-(((methylsulfonyl)oxy)methyl)-5-oxopyrrolidine-2-carboxylate (50 g, 111 mmol), isoindoline-1,3-dione (16.4 g, 112 mmol) and K2CO3 (25.1 g, 181 mmol) in NMP (400 mL) was stirred at 100° C. for 12 h. Upon completion, the reaction mixture was diluted with water (1 L) and filtered. The filter cake was dissolved with EtOAc (1 L), washed with brine (3×1 L), dried over Na2SO4 and evaporated. The crude product was triturated (PE/MTBE=1/1, 3.0 L) at 25° C. for 12 h to give tert-butyl (2S,3R)-3-((1,3-dioxoisoindolin-2-yl)methyl)-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-5-oxopyrrolidine-2-carboxylate (25 g, 44% yield) as a white solid.
m/z ES+ [M+H]+ 504.1; 1H NMR (400 MHz, CDCl3) δ ppm 8.46 (s, 1H), 7.92-7.85 (m, 2H), 7.80-7.73 (m, 2H), 7.09 (s, 1H), 4.74 (d, J=3.2 Hz, 1H), 4.07-3.94 (m, 1H), 3.86-3.74 (m, 1H), 3.02-2.89 (m, 2H), 2.61-2.51 (m, 1H), 2.48 (s, 3H), 1.34 (s, 9H).
Step e. Note: The following process was conducted in 4 parallel batches.
To a solution of tert-butyl (2S,3R)-3-((1,3-dioxoisoindolin-2-yl)methyl)-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-5-oxopyrrolidine-2-carboxylate (25.0 g, 49.7 mmol) in MeOH (250 mL) was added hydrazine monohydrate (85% purity; 4.68 g, 79.5 mmol). The reaction mixture was stirred at 70° C. for 12 h under a N2 atmosphere. Upon completion, the reaction mixture was cooled to rt, filtered and the filtrate was evaporated. The resulting residues from the parallel batches were combined, diluted with water (1 L), extracted with EtOAc (2×1 L), washed with brine (1 L), dried over Na2SO4 and evaporated to give tert-butyl (2S,3R)-3-(aminomethyl)-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-5-oxopyrrolidine-2-carboxylate (76 g, crude) as a brown gum.
m/z ES+ [M+H]+ 374.0; 1H NMR (400 MHz, CDCl3) δ ppm 8.49 (s, 1H), 7.09 (s, 1H), 4.77 (d, J=3.2 Hz, 1H), 3.01-2.85 (m, 3H), 2.46 (s, 3H), 2.43-2.38 (m, 2H), 1.43 (s, 9H).
Step f. Note: The following process was conducted in 4 parallel batches.
A mixture of tert-butyl (2S,3R)-3-(aminomethyl)-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-5-oxopyrrolidine-2-carboxylate (40 g, 107 mmol), 1-chloro-2-fluoro-3-nitrobenzene (CAS: 2106-49-2; 17.8 g, 102 mmol) and NaHCO3 (18.0 g, 214 mmol) in EtOH (380 mL) and water (20 mL) was stirred at 105° C. for 12 h. Upon completion, the reaction mixture was evaporated, diluted with EtOAc (5 L) and washed with water (2 L). The organic layers from the parallel batches were combined, dried over Na2SO4, evaporated and purified by column chromatography (PE/EA=10/1) to give tert-butyl (2S,3R)-3-(((2-chloro-6-nitrophenyl)amino)methyl)-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-5-oxopyrrolidine-2-carboxylate (175 g, 77% yield) as a yellow oil.
m/z ES+ [M+H]+ 529.1; 1H NMR (400 MHz, CDCl3) δ ppm 8.41 (s, 1H), 7.93 (dd, J=7.6, 1.6 Hz, 1H), 7.47 (dd, J=7.6, 1.6 Hz, 1H), 7.03 (s, 1H), 6.78 (t, J=8.4 Hz, 1H), 4.76 (d, J=2.4 Hz, 1H), 3.68-3.63 (m, 1H), 3.48-3.42 (m, 1H), 2.97-2.90 (m, 1H), 2.61-2.52 (m, 1H), 2.47-2.38 (m, 4H), 1.36 (s, 9H).
Step g. To a solution of tert-butyl (2S,3R)-3-(((2-chloro-6-nitrophenyl)amino)methyl)-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-5-oxopyrrolidine-2-carboxylate (110 g, 208 mmol) in THF (1.1 L) was added 3% Pt—V/C (11 g, 1.26 mmol). The reaction mixture was stirred at 25° C. for 16 h under a H2 (30 psi) atmosphere. Upon completion, the reaction mixture was filtered and evaporated to give tert-butyl (2S,3R)-3-(((2-amino-6-chlorophenyl)amino)methyl)-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-5-oxopyrrolidine-2-carboxylate (102 g, 98% yield) as a brown oil.
m/z ES+ [M+H]+ 499.0; 1H NMR (400 MHz, CDCl3) δ ppm 8.48 (s, 1H), 7.10 (s, 1H), 6.91-6.76 (m, 2H), 6.68-6.61 (m, 1H), 4.88 (d, J=4.0 Hz, 1H), 3.29-3.22 (m, 1H), 3.17-3.10 (m, 1H), 3.07-2.98 (m, 1H), 2.68-2.53 (m, 2H), 2.50 (s, 3H), 1.43 (s, 9H).
Step h. Note: The following process was conducted in 3 parallel batches.
To a solution of tert-butyl (2S,3R)-3-(((2-amino-6-chlorophenyl)amino)methyl)-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-5-oxopyrrolidine-2-carboxylate (34.0 g, 68.2 mmol) in DCM (170 mL) was added HCl in 1,4-dioxane (4 M, 170 mL). The reaction mixture was stirred at 25° C. for 12 h. Upon completion, the reaction mixture was diluted with water (1 L). The pH of the reaction mixture was adjusted to pH 5 with solid NaOH and then evaporated to remove the DCM. The resulting residues from the parallel batches were combined, extracted with EtOAc (3×1 L), washed with brine (1 L), dried over Na2SO4 and evaporated to give (2S,3R)-3-[(2-amino-6-chloro-anilino)methyl]-1-[6-methyl-4-(trifluoromethyl)-2-pyridyl]-5-oxo-pyrrolidine-2-carboxylic acid (90 g, 99% yield) as a yellow oil.
m/z ES+ [M+H]+ 442.9; 1H NMR (400 MHz, DMSO-d6) δ ppm 8.39 (s, 1H), 7.40 (s, 1H), 6.74-6.66 (m, 1H), 6.61-6.51 (m, 2H), 4.92 (d, J=2.8 Hz, 1H), 3.14-3.07 (m, 1H), 3.03-2.95 (m, 1H), 2.87-2.78 (m, 1H), 2.70-2.61 (m, 1H), 2.53-2.52 (m, 1H), 2.47 (s, 3H).
Step i. Note: The following process was conducted in 2 parallel batches.
To a solution of (2S,3R)-3-(((2-amino-6-chlorophenyl)amino)methyl)-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-5-oxopyrrolidine-2-carboxylic acid (30.0 g, 67.8 mmol) and DIPEA (17.5 g, 136 mmol) in DCM (1800 mL) and EtOAc (600 mL) was added T3P (50% in EtOAc; 43.1 g, 67.8 mmol). The reaction mixture was stirred at 20° C. for 1 h. Upon completion, the reaction mixture was quenched with water (500 mL) and extracted with EtOAc (3×1 L). The organic layers from the parallel batches were combined, washed with brine (1 L), dried over Na2SO4 and evaporated. Purification by column chromatography (PE/EtOAc/NH4OH=1/1/0.002) gave the title compound (40 g, 67% yield) as an off-white solid.
m/z ES+ [M+H]+ 425.1; 1H NMR (400 MHz, CDCl3) δ ppm 8.32 (s, 1H), 7.80 (s, 1H), 7.49-7.43 (m, 1H), 7.23-7.14 (m, 2H), 7.06 (s, 1H), 4.75 (d, J=8.8 Hz, 1H), 3.84-3.72 (m, 1H), 3.67-3.34 (m, 1H), 2.87-2.73 (m, 3H), 2.44 (s, 3H), 2.36-2.21 (m, 1H).
Step a. To a solution of tert-butyl 3-oxoazetidine-1-carboxylate (CAS: 398489-26-4; 29.2 g, 170 mmol) and benzyl piperazine-1-carboxylate (CAS: 31166-44-6; 25 g, 114 mmol) in MeOH (500 mL) was added 4 Å molecular sieves (25 g) and acetic acid (6.8 g, 114 mmol). The mixture was stirred at rt for 12 h, and then NaBH3CN (21.4 g, 341 mmol) was added. The mixture was stirred at rt for 1 h. Upon completion, the mixture was filtered and evaporated. The crude product was purified by reverse phase flash chromatography (water (0.1% TFA)/MeCN). The product containing fractions were combined, basified to pH 8 with sat. aq. Na2CO3 and evaporated to remove MeCN and water. The resulting residue was extracted with EtOAc (3×500 mL). The combined layers were washed with brine (800 mL), dried over Na2SO4 and evaporated to give obtain benzyl 4-(1-(tert-butoxycarbonyl)azetidin-3-yl)piperazine-1-carboxylate (30 g, 67% yield) as a yellow solid.
m/z ES+ [M+H]+ 376.4; 1H NMR (400 MHz, CDCl3) δ ppm 7.40-7.30 (m, 5H), 5.14 (s, 2H), 3.99-3.90 (m, 2H), 3.82 (s, 2H), 3.57 (s, 4H), 3.09 (s, 1H), 2.22 (s, 4H), 1.43 (s, 9H).
Step b. A mixture of benzyl 4-(1-(tert-butoxycarbonyl)azetidin-3-yl)piperazine-1-carboxylate (60 g, 160 mmol) and 10% Pd/C (9 g) in MeOH (600 mL) was stirred at 40° C. for 16 h under a H2 atmosphere (50 psi). Upon completion, the mixture was filtered and evaporated to give the title compound (37 g, crude) as a white solid.
1H NMR (400 MHz, CDCl3) δ ppm 3.93-3.87 (m, 2H), 3.79 (m, 2H), 3.09-3.01 (m, 1H), 2.92-2.89 (m, 4H), 2.32 (s, 4H), 1.41 (s, 9H).
Step a. To a solution of tert-butyl 1,4-diazepane-1-carboxylate (CAS: 112275-50-0; 10.0 g, 49.9 mmol) in THF (100 mL) and water (30 mL) was added CbzCl (8.52 g, 49.9 mmol) and Na2CO3 (15.9 g, 150 mmol). The mixture was stirred at 25° C. for 2 h. Upon completion, the reaction mixture was quenched with sat. aq. NH4Cl (100 mL), diluted with water (100 mL) and extracted with EtOAc (3×200 mL). The combined organic layers were washed with brine (3×100 mL), dried over Na2SO4 and evaporated. The residue was purified by column chromatography (PE/EtOAc=4/1) to give 1-benzyl 4-(tert-butyl) 1,4-diazepane-1,4-dicarboxylate (15 g, 89% yield) as a yellow oil.
1H NMR (400 MHz, DMSO-d6) δ ppm 7.38-7.31 (m, 5H), 5.11-5.01 (m, 2H), 3.51-3.48 (m, 2H), 3.45-3.36 (m, 4H), 3.31-3.28 (m, 2H), 1.74-1.57 (m, 2H), 1.41-1.30 (m, 9H).
Step b. To a solution of 1-benzyl 4-(tert-butyl) 1,4-diazepane-1,4-dicarboxylate (5 g, 15.0 mmol) in DCM (50 mL) was added TFA (10 mL) and the mixture was stirred at 25° C. for 30 min. Upon completion, the reaction mixture was evaporated to give benzyl 1,4-diazepane-1-carboxylate as a TFA salt (5.1 g, crude) as a colourless oil.
m/z ES+ [M+H]+ 235.2.
Step c. To a solution of benzyl 1,4-diazepane-1-carboxylate (5.1 g, 14.6 mmol) and tert-butyl 3-oxoazetidine-1-carboxylate (3.01 g, 17.6 mmol) in MeOH (50 mL) was added acetic acid (4.40 g, 73.2 mmol), 4 Å molecular sieves (5 g) and NaBH3CN (1.84 g, 29.3 mmol). The mixture was stirred at 25° C. for 2 h. Upon completion, the reaction mixture was filtered and evaporated. The residue was diluted with sat. aq. NaHCO3 (50 mL) and water (100 mL), and extracted with EtOAc (3×150 mL). The combined organic layers were washed with brine (3×100 mL), dried over Na2SO4, filtered and evaporated. The residue was purified by column chromatography (PE/EtOAc=3/1) to give benzyl 4-(1-(tert-butoxycarbonyl)azetidin-3-yl)-1,4-diazepane-1-carboxylate (3.5 g, 61% yield) as a colourless oil.
m/z ES+ [M+H]+ 390.2. 1H NMR (400 MHz, CDCl3) δ ppm 7.30-7.40 (m, 5H), 5.14 (s, 2H), 4.60-4.57 (m, 4H), 3.98-3.87 (m, 2H), 3.77-3.72 (m, 2H), 3.59-3.51 (m, 4H), 3.31-3.25 (m, 1H), 2.38 (d, J=4.8 Hz, 2H), 1.66 (s, 9H).
Step d. To a solution of benzyl 4-(1-(tert-butoxycarbonyl)azetidin-3-yl)-1,4-diazepane-1-carboxylate (3.5 g, 8.99 mmol) in MeOH (50 mL) was added 10% Pd/C (954 mg) and the mixture was stirred at 25° C. for 1 h under a H2 atmosphere (15 psi). Upon completion, the reaction mixture was filtered and evaporated to give the title compound (2.1 g, 91% yield) as a colourless oil.
1H NMR (400 MHz, DMSO-d6) δ ppm 5.63 (s, 1H), 4.37-4.35 (m, 1H), 3.98 (t, J=7.6 Hz, 2H), 3.59-3.55 (m, 2H), 3.32-3.23 (m, 2H), 2.78-2.65 (m, 1H), 2.54-2.50 (m, 4H), 2.45-2.30 (m, 1H), 1.37-1.35 (m, 11H).
Step a. To a solution of tert-butyl (R)-3-(hydroxymethyl)piperazine-1-carboxylate (CAS: 278788-66-2; 5.00 g, 23.1 mmol) and benzaldehyde (2.94 g, 27.7 mmol) in DCE (50 mL) was added acetic acid (2.08 g, 34.6 mmol) and 4 Å molecular sieves (5 g). The mixture was stirred at 25° C. for 1 h, and then NaBH(OAc)3 (5.88 g, 27.7 mmol) was added. The mixture was stirred at 25° C. for 12 h. Upon completion, the mixture was filtered, diluted with water (20 mL) extracted with EtOAc (3×20 mL). The combined layers were dried over Na2SO4 and evaporated. The residue was purified by column chromatography (PE/EtOAc=3/1) to give tert-butyl (R)-4-benzyl-3-(hydroxymethyl)piperazine-1-carboxylate (3.5 g, 47% yield) as a yellow solid.
m/z ES+ [M+H]+ 307.2.
Step b. To a solution of tert-butyl (R)-4-benzyl-3-(hydroxymethyl)piperazine-1-carboxylate (500 mg, 1.63 mmol) in DCM (10 mL) was added DAST (1.32 g, 8.16 mmol) dropwise at 0° C. The mixture was stirred at 25° C. for 5 h. Upon completion, the mixture was diluted with sat. aq. NaHCO3 (20 mL) and extracted with EtOAc (2×30 mL). The combined organic layers were dried over Na2SO4 and evaporated. The residue was purified by column chromatography (PE/EtOAc=20/1) to give an inseparable mixture of tert-butyl (R)-4-benzyl-6-fluoro-1,4-diazepane-1-carboxylate and tert-butyl (R)-4-benzyl-3-(fluoromethyl)piperazine-1-carboxylate (450 mg, 86% yield) as a brown oil.
m/z ES+ [M+H]+ 309.3.
Step c. A mixture of tert-butyl (R)-4-benzyl-6-fluoro-1,4-diazepane-1-carboxylate and tert-butyl (R)-4-benzyl-3-(fluoromethyl)piperazine-1-carboxylate (400 mg, 1.30 mmol), 10% Pd/C (100 mg), and 20% Pd(OH)2/C (100 mg) in MeOH (10 mL) was stirred at 40° C. for 24 h under a H2 atmosphere (15 psi). Upon completion, the reaction mixture was filtered and evaporated to give an inseparable mixture of tert-butyl (R)-6-fluoro-1,4-diazepane-1-carboxylate and tert-butyl (R)-3-(fluoromethyl)piperazine-1-carboxylate (280 mg, 99% yield) as a colourless oil.
m/z ES+ [M+H]+ 219.1.
Step d. To a solution of tert-butyl (R)-6-fluoro-1,4-diazepane-1-carboxylate and tert-butyl (R)-3-(fluoromethyl)piperazine-1-carboxylate (280 mg, 1.28 mmol) and benzyl 3-oxoazetidine-1-carboxylate (395 mg, 1.92 mmol) in MeOH (5 mL) was added 4 Å molecular sieves (280 mg) and acetic acid (385 mg, 6.41 mmol). The mixture was stirred at 25° C. for 30 min, then NaBH3CN (121 mg, 1.92 mmol) was added. The mixture was stirred at 25° C. for 12 h. Upon completion, the reaction mixture was diluted with DCM (5 mL), basified to pH 8 with sat. aq. NaHCO3. The organic layer was separated and the aqueous was further extracted with DCM (2×5 mL). The combined organic layers were dried over Na2SO4 and evaporated. The residue was purified by column chromatography (PE/EtOAc=5/1) and then re-purified by Prep-TLC (PE/EtOAc=1/1) to give an inseparable mixture of tert-butyl (R)-4-(1-((benzyloxy)carbonyl)azetidin-3-yl)-6-fluoro-1,4-diazepane-1-carboxylate and tert-butyl (R)-4-(1-((benzyloxy)carbonyl)azetidin-3-yl)-3-(fluoromethyl)piperazine-1-carboxylate (250 mg, 37% yield) as a colourless oil.
m/z ES+ [M+H]+ 408.2.
Step e. To a solution of tert-butyl (R)-4-(1-((benzyloxy)carbonyl)azetidin-3-yl)-6-fluoro-1,4-diazepane-1-carboxylate and tert-butyl (R)-4-(1-((benzyloxy)carbonyl)azetidin-3-yl)-3-(fluoromethyl)piperazine-1-carboxylate (220 mg, 0.54 mmol) in DCM (3 mL) was added TFA (1 mL). The mixture was stirred at rt for 30 min. Upon completion, the mixture was diluted with DCM (5 mL) and basified to pH 8 with sat. aq. NaHCO3. The organic layer was separated and the aqueous was further extracted with DCM (2×5 mL). The combined organic layers were dried over Na2SO4 and evaporated to give an inseparable mixture of the title compounds (100 mg, crude) as a colourless oil.
m/z ES+ [M+H]+ 307.9.
Step a. To a solution of tert-butyl 3-oxoazetidine-1-carboxylate (100 g, 584 mmol) in MeNO2 (678 g, 11.1 mol) was added TEA (11.8 g, 117 mmol). The mixture was stirred at 25° C. for 1 h. Upon completion, the mixture was evaporated to give tert-butyl 3-hydroxy-3-(nitromethyl)azetidine-1-carboxylate (135 g, crude) as a yellow solid.
1H NMR (400 MHz, DMSO-d6) δ ppm 6.45 (s, 1H), 4.86 (s, 2H), 4.04 (d, J=8.8 Hz, 2H), 3.74 (d, J=9.2 Hz, 2H), 1.38 (s, 9H).
Step b. To a solution of tert-butyl 3-hydroxy-3-(nitromethyl)azetidine-1-carboxylate (50 g, 215 mmol) in DCM (1 L) was added TEA (43.6 g, 431 mmol). Then MsCl (24.7 g, 215 mmol) in DCM (500 mL) was added dropwise at −78° C. The mixture was stirred at −78° C. for 1.5 h. Methyl 2-aminoacetate hydrochloride (CAS: 34582-32-6; 54.0 g, 430 mmol) and TEA (43.6 g, 431 mmol) was dissolved in DCM (500 mL) and stirred for 10 min, and this solution was added into the reaction mixture at −78° C. The mixture was stirred at 25° C. for 16 h. Upon completion, the mixture was diluted with water (400 mL) and extracted with DCM (2×500 mL). The combined organic layers were washed with brine (300 mL), dried over Na2SO4 and evaporated. The crude product was purified by silica gel chromatography column (PE/EtOAc=5/1) to give tert-butyl 3-((2-methoxy-2-oxoethyl)amino)-3-(nitromethyl)azetidine-1-carboxylate (41.5 g, 63% yield) as a white solid.
m/z ES+ [M-tBu+H]+ 248.1; 1H NMR (400 MHz, CDCl3) δ ppm 4.70 (d, J=4.4 Hz, 2H), 3.97-3.90 (m, 2H), 3.89-3.85 (m, 2H), 3.76 (s, 3H), 3.48 (s, 2H), 1.45 (s, 9H).
Step c. To a solution of tert-butyl 3-((2-methoxy-2-oxoethyl)amino)-3-(nitromethyl)azetidine-1-carboxylate (20 g, 66.0 mmol) in MeOH (500 mL) was added Raney-Ni (20 g, 233 mmol) and the reaction mixture was stirred at 25° C. for 12 h under a H2 atmosphere (50 psi). Upon completion, the mixture was filtered and evaporated to give tert-butyl 7-oxo-2,5,8-triazaspiro[3.5]nonane-2-carboxylate (63 g, crude) as a colourless oil.
m/z ES+ [M+H]+ 242.1.
Step d. To a solution of tert-butyl 7-oxo-2,5,8-triazaspiro[3.5]nonane-2-carboxylate (63 g, 261 mmol) and NaHCO3 (65.8 g, 783 mmol) in THF (600 mL) and water (200 mL) was added CbzCl (66.8 g, 392 mmol) and the mixture was stirred at 25° C. for 12 h. Upon completion, the mixture was diluted with water (200 mL) and extracted with EtOAc (2×200 mL). The combined organic layers were washed with brine (200 mL), dried over Na2SO4 and evaporated. The crude product was purified by column chromatography (PE/EtOAc=1/1) to give 5-benzyl 2-(tert-butyl) 7-oxo-2,5,8-triazaspiro[3.5]nonane-2,5-dicarboxylate (46 g, 42% yield) as a colourless oil.
m/z ES+ [M+Na]+ 398.2; 1H NMR (400 MHz, CDCl3) δ ppm 7.37-7.23 (m, 5H), 6.49 (s, 1H), 5.08 (s, 2H), 4.21 (d, J=9.2 Hz, 2H), 4.10-4.06 (m, 2H), 3.73 (d, J=9.2 Hz, 2H), 3.56 (d, J=1.6 Hz, 2H), 1.37 (s, 9H).
Step e. To a solution of 5-benzyl 2-(tert-butyl) 7-oxo-2,5,8-triazaspiro[3.5]nonane-2,5-dicarboxylate (13 g, 34.6 mmol) in THF (100 mL) was added borane dimethyl sulfide complex (10 M in THF, 10.39 mL) at 0° C., and the mixture was stirred at 25° C. for 1 h. Upon completion, the mixture was quenched with MeOH (10 mL) and evaporated. The crude product was purified by column chromatography (PE/EtOAc=1/1) to give the title compound (2 g, 16% yield) as a yellow oil.
1H NMR (400 MHz, DMSO-d6) δ ppm 7.41-7.33 (m, 5H), 5.09 (s, 2H), 3.96 (d, J=9.2 Hz, 2H), 3.70 (d, J=8.8 Hz, 2H), 3.36-3.33 (m, 2H), 3.24-3.19 (m, 2H), 2.78 (s, 2H), 1.38 (s, 9H).
The title compound was prepared in a similar manner to Intermediate A4, using methyl D-alaninate (CAS: 14316-06-4) in step b.
m/z ES+ [M+H]+ 376.2.
The title compound was prepared in a similar manner to Intermediate A4, using methyl L-alaninate (CAS: 2491-20-5) in step b.
m/z ES+ [M-tBu+H]+ 320.2.
The title compound was prepared in a similar manner to Intermediate A4, using EtNO2 in step a.
m/z ES+ [M+H]+ 376.2.
The title compound was prepared in a similar manner to Intermediate A4, using EtNO2 in step a, and methyl D-alaninate in step b.
m/z ES+ [M+H]+ 390.3.
Step a. To a solution of Intermediate A5 (0.8 g, 2.13 mmol) in THF (9 mL) and water (3 mL) was added Na2CO3 (226 mg, 2.13 mmol) and CbzCl (545 mg, 3.20 mmol). The mixture was stirred at 25° C. for 12 h. Upon completion, the reaction mixture was quenched with sat. aq. NH4Cl (20 mL), further diluted with water (30 mL) and extracted with EtOAc (3×50 mL). The combined organic layers were washed with brine (3×20 mL), dried over Na2SO4 and evaporated. The residue was purified by column chromatography (PE/EtOAc=5/1) to give 5,8-dibenzyl 2-(tert-butyl) (R)-6-methyl-2,5,8-triazaspiro[3.5]nonane-2,5,8-tricarboxylate (1 g, 92% yield) as a colourless solid.
m/z ES+ [M+H]+ 510.4; 1H NMR (400 MHz, CDCl3) δ ppm 7.37-7.27 (m, 10H), 5.25-5.08 (m, 4H), 4.51-4.98 (m, 1H), 4.37-4.23 (m, 2H), 4.00-3.96 (m, 1H), 3.77-3.48 (m, 4H), 3.26-3.22 (d, J=13.2 Hz, 1H), 1.44 (s, 9H), 1.17 (d, J=6.0 Hz, 3H).
Step b. To a solution of 5,8-dibenzyl 2-(tert-butyl) (R)-6-methyl-2,5,8-triazaspiro[3.5]nonane-2,5,8-tricarboxylate (1.0 g, 1.96 mmol) in THF (10 mL) was added 10% Pd/C (208 mg) and the mixture was stirred at 25° C. for 12 h under a H2 atmosphere (15 psi). Upon completion, the reaction mixture was filtered and evaporated. The residue was purified by reverse phase flash chromatography (water (0.1% FA)/MeCN) to give the title compound (450 mg, 95% yield) as a colourless oil.
1H NMR (400 MHz, CDCl3) δ ppm 3.94 (d, J=8.8 Hz, 1H), 3.76-3.68 (m, 2H), 3.56 (d, J=9.2 Hz, 1H), 3.04 (d, J=11.8 Hz, 1H), 2.89-2.84 (m, 1H), 2.82-2.78 (m, 1H), 2.73-2.66 (m, 1H), 2.34-2.28 (m, 1H), 1.44 (s, 9H), 1.02 (d, J=6.0 Hz, 3H).
Step a. This step was conducted as described in Intermediate A4, step a.
Step b. To a solution of tert-butyl 3-hydroxy-3-(nitromethyl)azetidine-1-carboxylate (5.0 g, 21.5 mmol) and TEA (4.4 g, 43.1 mmol) in DCM (100 mL) was added MsCl (2.5 g, 21.5 mmol) at −78° C. under a N2 atmosphere. The mixture was stirred at −78° C. for 2 h. A solution of methyl D-prolinate hydrochloride (CAS: 184719-80-0; 5.6 g, 33.6 mmoll) and TEA (4.4 g, 43.1 mmol) in DCM (100 mL) was then added to the reaction mixture at −78° C. The reaction mixture was stirred at 25° C. for 12 h under a N2 atmosphere. Upon completion, the reaction mixture was diluted with DCM (100 mL), washed with brine (3×100 mL), dried over Na2SO4 and evaporated. The crude product was purified by column chromatography (PE/EtOAc=1/1) to give methyl (1-(tert-butoxycarbonyl)-3-(nitromethyl)azetidin-3-yl)-D-prolinate (5.0 g, 68% yield) as a yellow oil.
1H NMR (400 MHz, CDCl3) δ ppm 4.83-4.68 (m, 2H), 4.15-4.10 (m, 2H), 4.08-3.99 (m, 2H), 3.90-3.80 (m, 2H), 3.71 (s, 3H), 3.27-3.18 (m, 1H), 2.04-1.89 (m, 4H), 1.47 (s, 9H).
Step c. To a solution of methyl (1-(tert-butoxycarbonyl)-3-(nitromethyl)azetidin-3-yl)-D-prolinate (5.0 g, 14.6 mmol) in MeOH (100 mL) was added Raney-Ni (1.3 g, 14.6 mmol) and the mixture was at 25° C. for 12 h under a H2 atmosphere (50 psi). Upon completion, the reaction mixture was filtered and evaporated to give tert-butyl (R)-1′-oxohexahydro-1′H-spiro[azetidine-3,4′-pyrrolo[1,2-a]pyrazine]-1-carboxylate (3.5 g, 85% yield) as a yellow solid.
m/z ES+ [M+H]+ 282.2.
Step d. To a mixture of tert-butyl (R)-1′-oxohexahydro-1′H-spiro[azetidine-3,4′-pyrrolo[1,2-a]pyrazine]-1-carboxylate (400 mg, 1.42 mmol) in THF (5 mL) was added borane dimethyl sulfide complex (10 M in THF, 0.43 mL). The mixture was stirred at 25° C. for 1 h under a N2 atmosphere. Upon completion, the reaction mixture was quenched with MeOH (10 mL) at 0° C. and stirred at 25° C. for 2 h. The mixture was evaporated to give the title compound (400 mg, crude) as a white solid.
1H NMR (400 MHz, CDCl3) δ ppm 4.09-3.91 (m, 2H), 3.86-3.70 (m, 1H), 3.67-3.54 (m, 2H), 3.53-3.33 (m, 1H), 3.28-3.09 (m, 1H), 2.97-2.73 (m, 1H), 2.73-2.56 (m, 1H), 2.52-2.17 (m, 2H), 2.03-1.70 (m, 4H), 1.52-1.43 (m, 9H).
Step a. To a solution of N-(tert-butoxycarbonyl)-O-methyl-D-serine (CAS Number 86123-95-7; 500 mg, 2.28 mmol) in MeOH (5 mL) was added SOCl2 (597 mg, 5.02 mmol) dropwise at 0° C. The reaction mixture was stirred at 20° C. for 2 h. Upon completion, the mixture was evaporated. The residue was triturated with EtOAc (30 mL), filtered and the filter cake was dried to give methyl O-methyl-D-serinate hydrochloride (310 mg, 80% yield) as a white solid.
1H NMR (400 MHz, DMSO-d6) δ ppm 8.74 (br s, 3H), 4.27 (t, J=3.6 Hz, 1H), 3.78 (d, J=3.6 Hz, 2H), 3.74 (s, 3H), 3.29-3.26 (s, 3H).
Steps b-d. These 3 steps were conducted in a similar manner to Intermediate A10, steps b-d.
m/z ES+ [M+H]+ 271.9.
Step a. This step was conducted as described in Intermediate A4, step a.
Step b. A To a solution of tert-butyl 3-hydroxy-3-(nitromethyl)azetidine-1-carboxylate (2 g, 8.61 mmol) in DCM (20 mL) was added DAST (2.08 g, 12.9 mmol) dropwise at 0° C. The reaction mixture was stirred at 0° C. for 30 min. Upon completion, the mixture was quenched with water (20 mL), and the layers were separated. The organic layer was washed with brine (50 mL), dried over Na2SO4 and evaporated. The crude product was purified by column chromatography (PE/EtOAc=8/1) to give tert-butyl 3-(nitromethylene)azetidine-1-carboxylate (1.55 g, 84% yield) as a yellow solid.
1H NMR (400 MHz, CDCl3) δ ppm 7.08-7.02 (m, 1H), 5.06-5.00 (m, 2H), 4.75-4.67 (m, 2H), 1.48 (s, 9H).
Step c. To a solution of tert-butyl 3-(nitromethylene)azetidine-1-carboxylate (1.35 g, 6.30 mmol) in DCM (20 mL) was added methyl morpholine-3-carboxylate (CAS: 126264-49-1; 1.37 g, 9.45 mmol). The reaction mixture was stirred at 20° C. for 12 h. Upon completion, the mixture was evaporated and the crude product was purified by column chromatography (PE/EtOAc=5/1) to give methyl 4-(1-(tert-butoxycarbonyl)-3-(nitromethyl)azetidin-3-yl)morpholine-3-carboxylate (1.2 g, 53% yield) as yellow oil.
1H NMR (400 MHz, CDCl3) δ ppm 4.77-4.65 (m, 2H), 4.33 (d, J=8.8 Hz, 1H), 4.23 (d, J=10.8 Hz, 1H), 4.07 (d, J=9.6 Hz, 1H), 3.94 (d, J=9.6 Hz, 1H), 3.84 (dd, J=2.8, 10.8 Hz, 1H), 3.76 (s, 3H), 3.71 (d, J=8.8 Hz, 1H), 3.62 (dd, J=3.2, 11.2 Hz, 1H), 3.57-3.49 (m, 2H), 3.37-3.26 (m, 1H), 2.43 (d, J=11.8 Hz, 1H), 1.46 (s, 9H).
Steps d-e. These 2 steps were conducted in a similar manner to Intermediate A10, steps c-d.
m/z ES+ [M+H]+ 284.0.
The title compound was prepared in a similar manner to Intermediate A12, using nitroethane in step a, and methyl (R)-morpholine-3-carboxylate (CAS: 1187933-47-6) in step c. The two diastereoisomers of unknown absolute configuration were separated during step d using Prep-HPLC. The separated enantiomers were then used separately in step e.
Intermediate A13a
m/z ES+ [M+H]+ 298.0.
Intermediate A13b
m/z ES+ [M+H]+ 298.0.
Step a. A mixture of 1,4-bis(tert-butoxycarbonyl)piperazine-2-carboxylic acid (CAS: 181955-79-3; 5 g, 15.1 mmol), methyl L-alaninate hydrochloride (CAS: 2491-20-5; 2.53 g, 18.2 mmol), HATU (5.75 g, 15.1 mmol) and DIPEA (9.78 g, 75.7 mmol) in DCM (50 mL) was stirred at 25° C. for 1 h. Upon completion, the reaction mixture was evaporated and the crude product was purified by column chromatography (PE/EtOAc=1/1) to give di-tert-butyl 2-(((S)-1-methoxy-1-oxopropan-2-yl)carbamoyl)piperazine-1,4-dicarboxylate (6.0 g, 95% yield) as a colourless oil.
m/z ES+ [M+H]+ 416.5; 1H NMR (400 MHz, CDCl3) δ ppm 7.01-6.57 (m, 1H), 4.65-4.61 (m, 1H), 4.58-4.37 (m, 2H), 3.99-3.80 (m, 2H), 3.76-3.71 (m, 3H), 3.29-2.85 (m, 3H), 1.49 (s, 9H), 1.45 (s, 9H), 1.40 (d, J=7.2 Hz, 3H).
Step b. A mixture of di-tert-butyl 2-(((S)-1-methoxy-1-oxopropan-2-yl)carbamoyl)piperazine-1,4-dicarboxylate (2.0 g, 4.8 mmol) and TFA (15.4 g, 135 mmol) in DCM (10 mL) was stirred at 0° C. for 2 h. Upon completion, the reaction mixture was evaporated to give methyl (piperazine-2-carbonyl)-L-alaninate as a TFA salt (1.6 g, crude) as a yellow oil.
m/z ES+ [M+H]+ 216.1.
Step c. A mixture of methyl (piperazine-2-carbonyl)-L-alaninate (1.6 g, 4.86 mmol) and TEA (2.46 g, 24.3 mmol) in THF (20 mL) was stirred at 60° C. for 4 h. Upon completion, the reaction mixture was evaporated and the crude product was purified by column chromatography (DCM/MeOH=10/1) to give (7S)-7-methylhexahydro-2H-pyrazino[1,2-a]pyrazine-6,9-dione (1.0 g, crude) as a yellow oil.
m/z ES+ [M+H]+ 184.2; 1H NMR (400 MHz, CDCl3) δ ppm 4.74-4.60 (m, 1H), 4.50-4.45 (m, 1H), 4.23-4.14 (m, 2H), 3.85-3.79 (m, 1H), 3.42-3.30 (m, 1H), 3.07-2.85 (m, 3H), 1.55 (t, J=6.8 Hz, 3H).
Step d. A mixture of (7S)-7-methylhexahydro-2H-pyrazino[1,2-a]pyrazine-6,9-dione (1.0 g, 5.46 mmol), CbzCl (1.12 g, 6.55 mmol), NaHCO3 (2.29 g, 27.3 mmol) in THF (10 mL) and water (10 mL) was stirred at 25° C. for 12 h. Upon completion, the reaction mixture was evaporated, water (100 mL) was added and the mixture was extracted with EtOAc (3×50 mL). The combined organic layers were washed with brine (50 mL), dried over Na2SO4 and evaporated. The crude product was purified by column chromatography (PE/EtOAc=1/1) to give benzyl (7S)-7-methyl-6,9-dioxooctahydro-2H-pyrazino[1,2-a]pyrazine-2-carboxylate (550 mg, 32% yield) as a colourless oil.
1H NMR (400 MHz, CDCl3) δ ppm 7.38-7.23 (m, 5H), 6.54-6.41 (m, 1H), 5.22-5.02 (m, 2H), 4.64-4.62 (m, 1H), 4.48 (t, J=12.4 Hz, 1H), 4.24-4.02 (m, 2H), 3.98-3.85 (m, 1H), 2.83-2.78 (m, 2H), 2.72-2.56 (m, 1H), 1.46 (t, J=7.6 Hz, 3H).
Step e. A mixture of benzyl (7S)-7-methyl-6,9-dioxooctahydro-2H-pyrazino[1,2-a]pyrazine-2-carboxylate (500 mg, 1.58 mmol) and borane dimethyl sulfide complex (10 M in THF, 0.47 mL) in THF (5 mL) was stirred at 20° C. for 1 h. Upon completion, the reaction mixture was quenched with MeOH (10 mL) and stirred at 25° C. for 2 h. The reaction mixture was evaporated to give benzyl (7S)-7-methyloctahydro-2H-pyrazino[1,2-a]pyrazine-2-carboxylate (500 mg, crude) as a colourless oil.
m/z ES+ [M+H]+ 290.5.
Step f. A mixture of benzyl (7S)-7-methyloctahydro-2H-pyrazino[1,2-a]pyrazine-2-carboxylate (500 mg, 1.73 mmol), di-tert-butyl dicarbonate (377 mg, 1.73 mmol) and DIPEA (669 mg, 5.18 mmol) in THF (10 mL) was stirred at 25° C. for 1 h. Upon completion, the reaction mixture was evaporated and purified by column chromatography (PE/EtOAc=1/1) to give two diastereoisomers of unknown absolute configuration.
8-Benzyl 2-(tert-butyl) (3S,9aS/R)-3-methylhexahydro-2H-pyrazino[1,2-a]pyrazine-2,8(1H)-dicarboxylate (110 mg, 16% yield) as a yellow oil.
m/z ES+ [M+H]+ 390.1; 1H NMR (400 MHz, CDCl3) δ ppm 7.46-7.31 (m, 5H), 5.15 (s, 2H), 4.00-3.88 (m, 3H), 3.61-3.56 (m, 1H), 3.17-3.07 (m, 2H), 2.99-2.96 (m, 1H), 2.90-2.84 (m, 1H), 2.76-2.72 (m, 1H), 2.62-2.51 (m, 1H), 2.45-2.37 (m, 1H), 2.15 (dd, J=7.2, 11.6 Hz, 1H), 1.47 (s, 9H), 1.34 (d, J=6.4 Hz, 3H).
8-Benzyl 2-(tert-butyl) (3S,9aR/S)-3-methylhexahydro-2H-pyrazino[1,2-a]pyrazine-2,8(1H)-dicarboxylate (110 mg, 16% yield) as a yellow oil.
m/z ES+ [M+H]+ 390.1; 1H NMR (400 MHz, CDCl3) δ ppm 7.38-7.32 (m, 5H), 5.15 (s, 2H), 4.22-3.91 (m, 3H), 3.87-3.60 (m, 1H), 3.08 (s, 1H), 2.79-2.51 (m, 4H), 2.32-2.29 (m, 1H), 2.16-2.04 (m, 1H), 1.96-1.90 (m, 1H), 1.47 (s, 9H), 1.28-1.21 (m, 3H).
Step g. To a solution of 8-benzyl 2-(tert-butyl) (3S,9aR/S)-3-methylhexahydro-2H-pyrazino[1,2-a]pyrazine-2,8(1H)-dicarboxylate (100 mg, 0.26 mmol) in 2-propanol (10 mL) was added 10% Pd/C (10 mg) and the mixture was stirred at rt for 4 h under a H2 atmosphere (15 psi). Upon completion, the reaction mixture was filtered and evaporated to give tert-butyl (3S,9aR/S)-3-methyloctahydro-2H-pyrazino[1,2-a]pyrazine-2-carboxylate (50 mg, 76% yield) as a colourless oil.
Intermediate A14a:
m/z ES+ [M+H]+ 256.5.
Intermediate A14b was prepared in a similar manner to Intermediate A14a, using 8-benzyl 2-(tert-butyl) (3S,9aS/R)-3-methylhexahydro-2H-pyrazino[1,2-a]pyrazine-2,8(1H)-dicarboxylate in step g.
Intermediate A14b:
m/z ES+ [M+H]+ 256.5.
Step a. To a solution of benzyl 6-oxa-3-azabicyclo[3.1.0]hexane-3-carboxylate (CAS: 31865-25-5; 10.0 g, 45.6 mmol) in DMSO (40 mL) and water (40 mL) was added Cs2CO3 (22.3 g, 68.4 mmol) and tert-butyl 3-hydroxyazetidine-1-carboxylate (CAS: 141699-55-0; 9.5 g, 54.7 mmol). The reaction mixture was then stirred at 110° C. under microwave irradiation for 45 min. Upon completion, the reaction mixture was poured into cold water (50 mL) and extracted with EtOAc (3×100 mL). The combined organic layers were washed with cold water, brine, dried over Na2SO4 and evaporated. Purification by column chromatography gave benzyl (rel-trans)-3-((1-(tert-butoxycarbonyl)azetidin-3-yl)oxy)-4-hydroxypyrrolidine-1-carboxylate as an off-white solid (3.7 g, 21% yield).
m/z ES+ [M-CO2tBu]+ 293.6.
Step b. To a solution of benzyl (rel-trans)-3-((1-(tert-butoxycarbonyl)azetidin-3-yl)oxy)-4-hydroxypyrrolidine-1-carboxylate (3.7 g, 9.4 mmol) in MeOH (100 mL) was added 10% Pd/C (3.6 g) and the mixture was stirred at rt for 2 h under a H2 atmosphere. Upon completion, the reaction mixture was filtered and evaporated to give the title compound as an off-white solid (1.5 g, 62% yield).
m/z ES+ [M+H]+ 259.6.
Step a. To a solution of tert-butyl (rel-trans)-hexahydropyrrolo[3,4-b][1,4]oxazine-6(2H)-carboxylate (CAS: 138026-93-4; 950 mg, 4.16 mmol) and benzyl 3-oxoazetidine-1-carboxylate (CAS: 105258-93-3; 1.28 g, 6.24 mmol) in DCE (15 mL) was added 4 Å molecular sieves (1 g). The mixture was stirred at 20° C. for 30 min, then NaBH(OAc)3 (2.65 g, 12.8 mmol) was added. The mixture was stirred at 20° C. for a further 12 h. Upon completion, the mixture was filtered and evaporated. The residue was purified by column chromatography (PE/EtOAc=1/1) to give tert-butyl (rel-trans)-4-(1-((benzyloxy)carbonyl)azetidin-3-yl)hexahydropyrrolo[3,4-b][1,4]oxazine-6(2H)-carboxylate (1.8 g, crude) as a colourless oil.
1H NMR (400 MHz, CDCl3) δ ppm 7.36-7.32 (m, 5H), 5.11 (s, 2H), 4.68-4.61 (m, 1H), 4.27-4.23 (m, 1H), 4.05-3.99 (m, 2H), 3.90 (dd, J=4.4, 10.4 Hz, 2H), 3.86-3.76 (m, 1H), 3.65-3.54 (m, 2H), 3.31-3.19 (m, 1H), 3.14-3.03 (m, 2H), 2.79-2.67 (m, 1H), 2.29-2.12 (m, 2H), 1.46 (s, 9H).
Step b. To a solution of tert-butyl (rel-trans)-4-(1-((benzyloxy)carbonyl)azetidin-3-yl)hexahydropyrrolo[3,4-b][1,4]oxazine-6(2H)-carboxylate (1.0 g, 2.40 mmol) in DCM (10 mL) was added TFA (2 mL). The mixture was stirred at 20° C. for 30 min. Upon completion, the mixture was diluted with DCM (5 mL) and basified to pH 8 with sat. aq. NaHCO3. The layers were separated, and the aqueous phase was further extracted with DCM (4×15 mL). The combined organic layers were washed with brine (20 mL), dried over Na2SO4 and evaporated to give the title compound (760 mg, crude) as a colourless oil.
m/z ES+ [M+H]+ 318.2.
Step a. To a solution of tert-butyl 3-cyanopiperazine-1-carboxylate (CAS: 859518-35-7; 1.3 g, 6.15 mmol) in THF (10 mL) and water (10 mL) was added Na2CO3 (1.3 g, 12.3 mmol) and CbzCl (1.15 g, 6.77 mmol). The mixture was stirred at 25° C. for 12 h. Upon completion, the reaction mixture was diluted with H2O (20 mL) and extracted with EtOAc (3×30 mL). The combined organic layers were washed with brine (50 mL), dried over Na2SO4, evaporated and purified by prep-TLC (PE/EtOAc=6/1) to give 1-benzyl 4-(tert-butyl) 2-cyanopiperazine-1,4-dicarboxylate (100 mg, 43% yield) as an off-white solid.
m/z ES+ [M+Na]+ 368.1.
Step b. A mixture of 1-benzyl 4-(tert-butyl) 2-cyanopiperazine-1,4-dicarboxylate (2.1 g, 6.08 mmol), Raney-Ni (1 g, 11.7 mmol) and 30% NH4OH (71 mg, 0.61 mmol) in MeOH (25 mL) was degassed and purged with H2 3 times. The reaction mixture was stirred at 25° C. for 5 h under a H2 atmosphere (50 psi). Upon completion, the reaction mixture was filtered and evaporated to give 1-benzyl 4-(tert-butyl) 2-(aminomethyl)piperazine-1,4-dicarboxylate (2.1 g, crude) as a colourless oil.
1H NMR (400 MHz, CDCl3) δ ppm 7.36-7.28 (m, 5H), 5.22-5.11 (m, 2H), 4.15-3.84 (m, 4H), 3.12-2.80 (m, 5H), 1.47 (s, 9H), 1.07 (s, 2H).
Step c. To a solution of 1-benzyl 4-(tert-butyl) 2-(aminomethyl)piperazine-1,4-dicarboxylate (2.1 g, 6.01 mmol) in THF (25 mL) was added TEA (1.22 g, 12.0 mmol) and methyl 2-bromopropanoate (CAS: 5445-17-0; 1.30 g, 7.81 mmol). The reaction mixture was stirred at 80° C. for 12 h. Upon completion, the mixture was diluted with water (40 mL) and extracted with EtOAc (3×40 mL). The combine organic layers were washed with brine (100 mL), dried over Na2SO4, evaporated and purified by column chromatography (PE/EtOAc=1/1) to give 1-benzyl 4-(tert-butyl) 2-(((1-methoxy-1-oxopropan-2-yl)amino)methyl)piperazine-1,4-dicarboxylate (1.5 g, 57% yield) as a colourless oil.
m/z ES+ [M+H]+ 436.2; 1H NMR (400 MHz, CDCl3) δ ppm 7.39-7.31 (m, 5H), 5.20-5.09 (m, 2H), 4.23-4.06 (m, 2H), 4.02-3.83 (m, 2H), 3.70 (s, 3H), 3.44-3.27 (m, 1H), 3.09-2.90 (m, 2H), 2.80 (dd, J=12.0, 8.0 Hz, 2H), 2.56 (dd, J=11.6, 8.0 Hz, 1H), 1.67-1.53 (m, 1H), 1.47 (d, J=3.6 Hz, 9H), 1.30-1.20 (m, 3H).
Step d. A mixture of 1-benzyl 4-(tert-butyl) 2-(((1-methoxy-1-oxopropan-2-yl)amino)methyl)piperazine-1,4-dicarboxylate (2.9 g, 6.66 mmol), 10% Pd/C (300 mg) in MeOH (40 mL) was degassed and purged with H2 3 times. The reaction mixture was stirred at 45° C. for 12 h under a H2 atmosphere (15 psi). Upon completion, the reaction mixture was filtered and evaporated to give tert-butyl 7-methyl-6-oxooctahydro-2H-pyrazino[1,2-a]pyrazine-2-carboxylate (1.75 g, crude) as a colourless oil.
1H NMR. (400 MHz, CDCl3) δ ppm 4.64-4.54 (m, 1H), 4.11-3.91 (m, 2H), 3.57-3.32 (m, 2H), 3.31-3.13 (m, 1H), 2.98-2.69 (m, 3H), 2.68-2.45 (m, 2H), 1.48 (s, 9H), 1.40 (dd, J=6.8, 4.0 Hz, 3H).
Step e. To a solution of tert-butyl 7-methyl-6-oxooctahydro-2H-pyrazino[1,2-a]pyrazine-2-carboxylate (1.2 g, 4.46 mmol) in THF (12 mL) and water (12 mL) was added Na2CO3 (944 mg, 8.91 mmol) and CbzCl (1.14 g, 6.68 mmol). The reaction mixture was stirred at 25° C. for 12 h. Upon completion, the reaction mixture was diluted with water (20 mL) and extracted with EtOAc (3×30 mL). The combined organic layers were washed with brine (60 mL), dried over Na2SO4, evaporated and purified by column chromatography (PE/EtOAc=3/2) to give two products of unknown absolute configuration.
Intermediate A17a: 2-benzyl 8-(tert-butyl) (rel-3R/S,9aR/S)-3-methyl-4-oxohexahydro-2H-pyrazino[1,2-a]pyrazine-2,8(1H)-dicarboxylate (550 mg, 29% yield) as a white solid.
m/z ES+ [M-tBu+H]+ 348.0; 1H NMR (400 MHz, CDCl3) δ ppm 7.43-7.30 (m, 5H), 5.16 (s, 2H), 4.84-4.65 (m, 1H), 4.52 (d, J=12.4 Hz, 1H), 4.39-3.97 (m, 3H), 3.51-3.50 (m, 1H), 2.99-2.77 (m, 2H), 2.75-2.63 (m, 1H), 2.50-2.49 (m, 1H), 1.48 (s, 9H), 1.46 (d, J=7.2 Hz, 3H).
Intermediate A17b: 2-benzyl 8-(tert-butyl) (rel-3R/S,9aS/R)-3-methyl-4-oxohexahydro-2H-pyrazino[1,2-a]pyrazine-2,8(1H)-dicarboxylate (430 mg, 23% yield) as a yellow oil.
m/z ES+ [M-tBu+H]+ 348.0; 1H NMR (400 MHz, CDCl3) δ ppm 7.43-7.30 (m, 5H), 5.22-5.11 (m, 2H), 4.75 (d, J=6.0 Hz, 1H), 4.59 (d, J=11.4 Hz, 1H), 4.10-3.90 (m, 3H), 3.45 (dd, J=14.0, 4.8 Hz, 1H), 3.37 (d, J=11.6 Hz, 1H), 2.88-2.73 (m, 2H), 2.72-2.62 (m, 1H), 1.47 (s, 9H), 1.45 (d, J=7.2 Hz, 3H).
The title compounds were prepared by chiral SFC separation of Intermediate.
Intermediate A18a:
m/z ES+ [M+H]+ 404.0; 1H NMR (400 MHz, CDCl3) δ ppm 7.40-7.30 (m, 5H), 5.16 (s, 2H), 4.84-4.63 (m, 1H), 4.52 (d, J=12.4 Hz, 1H), 4.39-4.01 (m, 3H), 3.51-3.50 (m, 1H), 3.00-2.78 (m, 2H), 2.74-2.62 (m, 1H), 2.51-2.40 (m, 1H), 1.48 (s, 9H), 1.46 (d, J=7.2 Hz, 3H).
Intermediate A18b:
m/z ES+ [M-tBu]+ 347.9; 1H NMR (400 MHz, CDCl3) δ ppm 7.40-7.30 (m, 5H), 5.16 (s, 2H), 4.85-4.63 (m, 1H), 4.52 (d, J=12.4 Hz, 1H), 4.39-4.02 (m, 3H), 3.51-3.49 (s, 1H), 2.99-2.78 (m, 2H), 2.74-2.63 (m, 1H), 2.50-2.48 (s, 1H), 1.48 (s, 9H), 1.46 (d, J=7.2 Hz, 3H).
The title compounds were prepared by chiral SFC separation of Intermediate A17b.
Intermediate A18c:
m/z ES+ [M+H]+ 404.0.
Intermediate A18d:
m/z ES+ [M+H]+ 404.0.
Step a. To a solution of Intermediate A18a (110 mg, 0.27 mmol) in DCM (1.5 mL) was added TFA (508 mg, 4.46 mmol). The mixture was stirred at 25° C. for 1 h. Upon completion, the reaction mixture was basified to pH 8 with sat. aq. NaHCO3 and extracted with EtOAc (3×15 mL). The combine organic layers were washed with brine (30 mL), dried over Na2SO4 and evaporated to give the title compound (68 mg, crude) as a yellow solid.
m/z ES+ [M+H]+ 304.0
The title compound was prepared in a similar manner to Intermediate A19a, using Intermediate A18b.
m/z ES+ [M+H]+ 304.1.
The title compound was prepared in a similar manner to Intermediate A19a, using Intermediate A18c.
m/z ES+ [M+H]+ 304.1.
The title compound was prepared in a similar manner to Intermediate A19a, using Intermediate A18d.
m/z ES+ [M+H]+ 304.2.
Step a. A mixture of Intermediate A18a (110 mg, 0.27 mmol) and 10% Pd/C (10 mg) in MeOH (2 mL) was degassed and purged with H2 3 times. The mixture was stirred at 30° C. for 1 h under a H2 atmosphere (15 psi). The mixture was filtered and evaporated to give the title compound (65 mg, crude) as a yellow solid.
m/z ES+ [M+H]+ 270.0.
The title compound was prepared in a similar manner to Intermediate A20a, using Intermediate A18b.
m/z ES+ [M+H]+ 270.1.
The title compound was prepared in a similar manner to Intermediate A20a, using Intermediate A18c.
m/z ES+ [M+H]+ 270.1.
The title compound was prepared in a similar manner to Intermediate A20a, using Intermediate A18d.
m/z ES+ [M+H]+ 270.1.
Steps a-d. These 4 steps were conducted in a similar manner to Intermediate A17a and Intermediate A17b, steps a-d, using methyl 2-bromoacetate in step c.
m/z ES+ [M+H]+ 256.2; 1H NMR (400 MHz, CDCl3) δ ppm 4.62 (d, J=13.2 Hz, 1H), 4.06 (s, 2H), 3.61-3.46 (m, 2H), 3.45-3.36 (m, 1H), 3.30-3.20 (m, 1H), 2.83-2.78 (m, 1H), 2.77-2.70 (m, 1H), 2.69-2.53 (m, 2H), 1.48 (s, 9H).
Step a. This step was conducted in a similar manner to Intermediate A17a and Intermediate A17b, step e, using Intermediate A21.
Step b. This step was conducted in a similar manner to Intermediate 19a.
m/z ES+ [M+H]+ 290.1; 1H NMR (400 MHz, CDCl3) δ ppm 7.40-7.30 (m, 5H), 5.16 (s, 2H), 4.58 (d, J=10.8 Hz, 1H), 4.41 (d, J=18.0 Hz, 1H), 4.09 (s, 1H), 3.97 (d, J=18.0 Hz, 1H), 3.55-3.45 (m, 1H), 3.21-2.96 (m, 3H), 2.78-2.63 (m, 2H), 2.49 (t, J=11.2 Hz, 1H).
Step a. To a mixture of 6-methylpyrazine-2-carboxylic acid (10.0 g, 72.4 mmol) and Cs2CO3 (47.1 g, 144 mmol) in DMF (100 mL) was added MeI (15.4 g, 108 mmol). The reaction mixture was stirred at 25° C. for 1 h. Upon completion, the reaction mixture was quenched with water (100 mL) and extracted with EtOAc (3×150 mL). The combined organic layers were washed with brine (100 mL), dried over Na2SO4 and evaporated to give methyl 6-methylpyrazine-2-carboxylate (6.5 g, 59% yield) as a yellow solid.
1H NMR (400 MHz, CDCl3) δ ppm 9.15 (s, 1H), 8.67 (s, 1H), 4.06 (s, 3H), 2.71 (s, 3H).
Step b. A mixture of methyl 6-methylpyrazine-2-carboxylate (6.50 g, 42.7 mmol), PtO2 (650 mg, 2.86 mmol) and acetic acid (5.13 g, 85.4 mmol) in MeOH (120 mL) was stirred at 50° C. for 48 h under a H2 atmosphere (50 psi). Upon completion, the reaction mixture was filtered and evaporated to give methyl 6-methylpiperazine-2-carboxylate (6.0 g, 88% yield) as a yellow oil.
m/z ES+ [M+H]+ 158.9.
Step c. A mixture of methyl 6-methylpiperazine-2-carboxylate (5.0 g, 15.8 mmol) and benzaldehyde (1.68 g, 15.8 mmol) in MeOH (100 mL) was stirred at 0° C. for 30 min. After which, NaBH3CN (1.19 g, 18.9 mmol) was added and stirred at 0° C. for a further 1 h. Upon completion, the reaction mixture was evaporated, dissolved in water (50 mL) and extracted with EtOAc (2×50 mL). The combined organic layers were washed with brine (50 mL), dried over Na2SO4 and evaporated. The residue was purified by reverse phase flash chromatography (water (0.1% NH4OH)/MeCN) to give two products.
Intermediate A23a: methyl (rel-2R,6R)-4-benzyl-6-methylpiperazine-2-carboxylate (2.10 g, 53% yield) as yellow oil
m/z ES+ [M+H]+ 249.1; 1H NMR (400 MHz, CDCl3) δ ppm 7.43-7.28 (m, 5H), 3.74 (s, 3H), 3.64 (s, 1H), 3.59-3.51 (m, 1H), 3.47-3.40 (m, 1H), 3.29-3.15 (m, 2H), 2.70 (d, J=10.6 Hz, 1H), 2.34 (dd, J=11.2, 3.2 Hz, 1H), 1.86 (t, J=10.4 Hz, 1H), 1.04 (d, J=6.4 Hz, 3H).
Intermediate A23b: methyl (rel-2S,6R)-4-benzyl-6-methylpiperazine-2-carboxylate (1.20 g, 31% yield) as yellow oil.
m/z ES+ [M+H]+ 249.1; 1H NMR (400 MHz, CDCl3) δ ppm 7.41-7.28 (m, 5H), 3.72 (s, 3H), 3.60-3.55 (m, 2H), 3.17-3.14 (m, 1H), 3.00-2.85 (m, 1H), 2.80-2.75 (m, 1H), 2.10-1.95 (m, 1H), 1.75-1.67 (m, 2H), 1.07 (d, J=6.4 Hz, 3H).
Step a. A mixture of (tert-butoxycarbonyl)glycine (1.69 g, 9.66 mmol), HATU (3.67 g, 9.66 mmol) and DIPEA (3.12 g, 24.1 mmol) in DMF (15 mL) was stirred at 25° C. for 30 min. Intermediate A23a (2.00 g, 8.05 mmol) in DMF (15 mL) was then added and the reaction mixture was stirred at 25° C. for a further 2 h. Upon completion, the reaction mixture was diluted with water (50 mL) and extracted with EtOAc (2×50 mL). The combined organic layers were washed with brine (20 mL), dried over Na2SO4, evaporated and purified by column chromatography (PE/EtOAc=3/1) to give methyl (rel-2R,6R)-4-benzyl-1-((tert-butoxycarbonyl)glycyl)-6-methylpiperazine-2-carboxylate (2.3 g, 70% yield) as a yellow oil.
m/z ES+ [M+H]+ 406.2; 1H NMR (400 MHz, CDCl3) δ ppm 7.37-7.30 (m, 5H), 5.55 (s, 1H), 5.17-5.09 (m, 1H), 4.26-4.16 (m, 1H), 4.07-3.97 (m, 1H), 3.95-3.82 (m, 1H), 3.79-3.71 (m, 3H), 3.68-3.46 (m, 3H), 2.75-2.58 (m, 1H), 2.31-2.17 (m, 2H), 1.47 (s, 9H), 1.36 (d, J=6.4 Hz, 2H), 1.23 (d, J=4.4 Hz, 1H).
Step b. To a solution of methyl (rel-2R,6R)-4-benzyl-1-((tert-butoxycarbonyl)glycyl)-6-methylpiperazine-2-carboxylate (2.0 g, 4.93 mmol) in MeOH (20 mL) was added 4 M HCl in MeOH (10 mL) and the reaction mixture was stirred at 25° C. for 16 h. Upon completion, the mixture was evaporated to give methyl (rel-2R,6R)-4-benzyl-1-glycyl-6-methylpiperazine-2-carboxylate hydrochloride (1.50 g, 88% yield) as a white solid.
m/z ES+ [M+H]+ 306.0.
Step c. A mixture of methyl (rel-2R,6R)-4-benzyl-1-glycyl-6-methylpiperazine-2-carboxylate hydrochloride (1.50 g, 4.91 mmol) and TEA (2.49 g, 24.5 mmol) in MeOH (30 mL) was stirred at 70° C. for 2 h. Upon completion, the reaction mixture was evaporated and triturated with THF (5 mL) to give (rel-4R,9aR)-2-benzyl-4-methylhexahydro-2H-pyrazino[1,2-a]pyrazine-6,9-dione (1.20 g, 89% yield) as a yellow solid.
m/z ES+ [M+H]+ 274.0; 1H NMR (400 MHz, CDCl3) δ ppm 7.31-7.20 (m, 5H), 6.02 (s, 1H), 4.12-3.95 (m, 1H), 3.88 (s, 2H), 3.69-3.47 (m, 3H), 3.38-3.26 (m, 1H), 2.84-2.81 (m, 1H), 2.26-2.25 (m, 1H), 2.13-2.05 (m, 1H), 1.45 (d, J=6.4 Hz, 3H).
Step d. To a solution of (rel-4R,9aR)-2-benzyl-4-methylhexahydro-2H-pyrazino[1,2-a]pyrazine-6,9-dione (400 mg, 1.46 mmol) in THF (1 mL) was added LiAlH4 (555 mg, 14.6 mmol) and the reaction mixture was stirred at 0° C. for 1 h. Upon completion, the reaction mixture was slowly quenched with water (1 mL), 15% aq. NaOH solution (3 mL) and additional water (1 mL) at 0° C. and then Na2SO4 was added. The mixture was filtered and the filtrate was evaporated to give the title compound (330 mg, 91% yield) as yellow oil.
m/z ES+ [M+H]+ 246.1; 1H NMR (400 MHz, CDCl3) δ ppm 7.36-7.30 (m, 5H), 3.76-3.69 (m, 1H), 3.53-3.45 (m, 2H), 3.16-3.09 (m, 1H), 3.08-3.00 (m, 1H), 2.96-2.87 (m, 1H), 2.82-2.75 (m, 2H), 2.70-2.62 (m, 1H), 2.60-2.51 (m, 1H), 2.39-2.32 (m, 1H), 2.30-2.24 (m, 1H), 2.03-1.94 (m, 2H), 1.91-1.85 (m, 1H), 1.04 (d, J=6.4 Hz, 3H).
The title compound was prepared in a similar manner to Intermediate A24a, using Intermediate A23b in step a.
m/z ES+ [M+H]+ 246.1; 1H NMR 1H NMR (400 MHz, CDCl3) δ ppm 7.27-7.16 (m, 5H), 3.68-3.63 (m, 1H), 3.46-3.43 (m, 1H), 3.36-3.30 (m, 1H), 2.93-2.82 (m, 3H), 2.73 (dd, J=11.6, 1.6 Hz, 1H), 2.67-2.46 (m, 6H), 2.42-2.35 (m, 1H), 2.29 (dd, J=10.8, 3.2 Hz, 1H), 1.10 (d, J=6.4 Hz, 3H).
Step a. To a mixture of 1-benzhydryl-3-methyl-azetidin-3-ol (CAS: 40320-63-6; 500 mg, 1.97 mmol) and TEA (399 mg, 3.95 mmol) in DCM (10 mL) was added MsCl (339 mg, 2.96 mmol) at 0° C. The reaction mixture was stirred at 20° C. for 1 h under a N2 atmosphere. Upon completion, the mixture was diluted with water (50 mL) and extracted with EtOAc (3×20 mL). The combined organic layers were washed with brine (3×50 mL), dried over Na2SO4 and evaporated to give 1-benzhydryl-3-methylazetidin-3-yl methanesulfonate (650 mg, 99% yield) as a yellow solid.
1H NMR (400 MHz, CDCl3) δ ppm 7.42-7.37 (m, 4H), 7.31-7.25 (m, 4H), 7.24-7.16 (m, 2H), 4.44 (s, 1H), 3.35 (s, 4H), 3.04 (s, 3H), 1.92 (s, 3H).
Step b. A mixture of 1-benzhydryl-3-methylazetidin-3-yl methanesulfonate (1.0 g, 3.02 mmol) and piperazine (2.60 g, 30.1 mmol) in 2-propanol (10 mL) was stirred at 70° C. for 16 h. Upon completion, the reaction mixture was evaporated and purified by reverse phase flash chromatography (water (0.1% NH4OH)/MeCN) to give the title compound (950 mg, 97% yield) as a yellow solid.
m/z ES+ [M+H]+ 322.2; 1H NMR (400 MHz, CDCl3) δ ppm 7.48-7.37 (m, 4H), 7.32-7.28 (m, 4H), 7.23-7.17 (m, 2H), 4.44 (s, 1H), 3.13-3.06 (m, 6H), 2.86 (d, J=6.8 Hz, 2H), 2.58-2.51 (m, 4H), 1.38 (s, 3H).
Step a. To a solution of tert-butyl 3-oxoazetidine-1-carboxylate (5.0 g, 29.2 mmol) in THF (50 mL) was added Ti(OEt)4 (20 g, 87.6 mmol) and 2-methylpropane-2-sulfinamide (4.25 g, 35.1 mmol). The reaction mixture was stirred at 25° C. for 12 h. Upon completion, the mixture was diluted with EtOAc (300 mL) and quenched with water (150 mL) and filtered. The filtrate was extracted with EtOAc (3×150 mL) and the combined organic layers were washed with brine (150 mL), dried over Na2SO4, evaporated and purified by column chromatography (PE/EtOAc=5/1) to give tert-butyl 3-((tert-butylsulfinyl)imino)azetidine-1-carboxylate (7.5 g, 93.6% yield) as a yellow solid.
m/z ES+ [M+H]+ 275.2; 1H NMR (400 MHz, CDCl3) δ ppm 5.16-5.06 (m, 1H), 5.00-4.90 (m, 1H), 4.74 (t, J=2.4 Hz, 2H), 1.45 (s, 9H), 1.25 (s, 9H).
Step b. To a solution of tert-butyl 3-((tert-butylsulfinyl)imino)azetidine-1-carboxylate (7.1 g, 25.9 mmol) in DCM (100 mL) was added Ti(OEt)4 (11.8 g, 51.8 mmol) and trimethylsilanecarbonitrile (6.42 g, 64.7 mmol). The reaction mixture was stirred at 25° C. for 12 h. Upon completion, the mixture was quenched with water (200 mL) and extracted with DCM (300 mL and 3×200 mL). The combined organic layers were washed with brine (3×100 mL), dried over Na2SO4, evaporated and purified by column chromatography (PE/EtOAc=1/1) to give tert-butyl 3-((tert-butylsulfinyl)amino)-3-cyanoazetidine-1-carboxylate (6.6 g, 84% yield) as a yellow solid.
m/z ES+ [M+Na]+ 324.2; 1H NMR (400 MHz, CDCl3) δ ppm 4.50-4.38 (m, 2H), 4.31-4.24 (m, 1H), 4.16-4.14 (m, 1H), 4.04 (s, 1H), 1.46 (s, 9H), 1.29 (s, 9H).
Step c. To a solution of tert-butyl 3-((tert-butylsulfinyl)amino)-3-cyanoazetidine-1-carboxylate (2.0 g, 6.64 mmol) in THF (25 mL) and water (5 mL) was added iodine (337 mg, 1.33 mmol) and the mixture was stirred at 50° C. for 12 h. Upon completion, the reaction mixture was quenched with sat. aq. NaHCO3 (20 mL), diluted with water (30 mL) and extracted with EtOAc (3×50 mL). The combined organic layers were washed with brine (20 mL), dried over Na2SO4, evaporated and purified by column chromatography (PE/EtOAc=1/1) to give tert-butyl 3-amino-3-cyanoazetidine-1-carboxylate (950 mg, 73% yield) as a yellow solid.
1H NMR (400 MHz, CDCl3) δ ppm 4.35 (d, J=8.8 Hz, 2H), 3.89 (d, J=8.8 Hz, 2H), 2.03 (brs, 2H), 1.45 (s, 9H).
Step d. A mixture of tert-butyl 3-amino-3-cyanoazetidine-1-carboxylate (300 mg, 1.52 mmol), CbzCl (389 mg, 2.28 mmol), and Na2CO3 (484 mg, 4.56 mmol) in THF (6 mL) and water (2 mL) was stirred at 25° C. for 2 h. Upon completion, the reaction was quenched with sat. aq. NH4Cl (20 mL), diluted with water (20 mL) and extracted with EtOAc (3×30 mL). The combined organic layers were washed with brine (20 mL), dried over Na2SO4, evaporated and purified by column chromatography (PE/EtOAc=5/1) to give tert-butyl 3-(((benzyloxy)carbonyl)amino)-3-cyanoazetidine-1-carboxylate (450 mg, 89% yield) as a yellow solid.
m/z ES+ [M+Na]+ 354.1; 1H NMR (400 MHz, CDCl3) δ ppm 7.38-7.30 (m, 5H), 5.66-5.46 (m, 1H), 5.18 (s, 2H), 4.43-4.41 (m, 2H), 4.11-4.00 (m, 2H), 1.45 (s, 9H).
Step e. To a solution of tert-butyl 3-(((benzyloxy)carbonyl)amino)-3-cyanoazetidine-1-carboxylate (300 mg, 0.91 mmol) in MeOH (30 mL) was added Raney-Ni (78 mg, 0.91 mmol) The suspension was degassed and purged with H2 3 times and the reaction mixture was stirred at 25° C. for 12 h under a H2 atmosphere (15 psi). Upon completion the reaction mixture was filtered and evaporated to give tert-butyl 3-(aminomethyl)-3-(((benzyloxy)carbonyl)amino)azetidine-1-carboxylate (300 mg, crude) as a yellow solid which was used in the next step directly.
m/z ES+ [M+H]+ 336.2.
Step f. To a solution of tert-butyl 3-(aminomethyl)-3-(((benzyloxy)carbonyl)amino)azetidine-1-carboxylate (300 mg, 0.90 mmol) in THF (5 mL) was added TEA (272 mg, 2.68 mmol) and methyl 2-bromoacetate (274 mg, 1.79 mmol). The reaction mixture was stirred at 25° C. for 12 h. Upon completion, the mixture was quenched with sat. aq. NH4Cl (20 mL), diluted with water (20 mL) and extracted with EtOAc (3×30 mL). The combined organic layers were washed with brine (20 mL), dried over Na2SO4, evaporated and purified by column chromatography (PE/EtOAc=1/2) to give tert-butyl 3-(((benzyloxy)carbonyl)amino)-3-(((2-methoxy-2-oxoethyl)amino)methyl)azetidine-1-carboxylate (300 mg, 82% yield) as a yellow solid.
m/z ES+ [M+H]+ 408.2; 1H NMR (400 MHz, CDCl3) δ ppm 7.41-7.29 (m, 5H), 5.81 (br s, 1H), 5.12 (s, 2H), 4.19-4.10 (m, 2H), 3.82 (d, J=8.8 Hz, 2H), 3.75 (s, 3H), 3.48 (s, 2H), 3.08 (s, 2H), 1.44 (s, 9H).
Step g. To a solution of tert-butyl 3-(((benzyloxy)carbonyl)amino)-3-(((2-methoxy-2-oxoethyl)amino)methyl)azetidine-1-carboxylate (1.3 g, 3.19 mmol) in MeOH (50 mL) was added 10% Pd/C (300 mg) and the mixture was stirred at 40° C. for 12 h under a H2 atmosphere (15 psi). Upon completion, the reaction mixture was filtered and evaporated to give the title compound (670 mg, crude) as a white solid.
1H NMR (400 MHz, DMSO-d6) δ ppm 8.27 (s, 1H), 3.81-3.69 (m, 4H), 3.11 (s, 2H), 2.90 (s, 2H), 2.75 (br s, 1H), 1.38 (s, 9H).
Step a. This step was conducted in a similar manner to Intermediate A26, step d, using Intermediate A26.
Step b. To a solution of 8-benzyl 2-(tert-butyl) 6-oxo-2,5,8-triazaspiro[3.5]nonane-2,8-dicarboxylate (0.25 g, 0.67 mmol) in DMF (5 mL) was added Cs2CO3 (651 mg, 2.0 mmol) and MeI (284 mg, 2.00 mmol). The reaction mixture was stirred at 25° C. for 12 h. Upon completion, the reaction mixture was quenched with sat. aq. NH4Cl (20 mL), diluted with water (30 mL) and extracted with EtOAc (3×30 mL). The combined organic layers were washed with brine (20 mL), dried over Na2SO4, evaporated and purified by column chromatography (PE/EtOAc=1/2) to give 8-benzyl 2-(tert-butyl) 5-methyl-6-oxo-2,5,8-triazaspiro[3.5]nonane-2,8-dicarboxylate (220 mg, 84% yield) as a colourless oil.
m/z ES+ [M+H]+ 390.1; 1H NMR (400 MHz, DMSO-d6) δ ppm 7.44-7.28 (m, 5H), 5.12 (s, 2H), 4.08 (d, J=9.2 Hz, 2H), 4.02 (s, 2H), 3.87 (s, 2H), 3.77 (d, J=9.6 Hz, 2H), 2.96 (s, 3H), 1.38 (s, 9H).
Step c. This step was conducted in a similar manner to Intermediate A26, step g.
m/z ES+ [M+H]+ 256.1.
Steps a-b. These 2 steps were conducted as described in Intermediate A12, steps a-b.
1H NMR (400 MHz, CDCl3) δ ppm 7.05 (s, 1H), 5.03 (d, J=3.2 Hz, 2H), 4.71 (s, 2H), 1.48 (s, 9H).
Step c. To a solution of methyl 1H-imidazole-2-carboxylate (2.59 g, 20.5 mmol) in toluene (40 mL) was added NaH (971 mg, 24.3 mmol, 60% dispersion in mineral oil). The mixture was stirred at 0° C. for 30 min, after which tert-butyl 3-(nitromethylene)azetidine-1-carboxylate (4 g, 18.7 mmol) in toluene (40 mL) was slowly added. The mixture was stirred at 0° C. for a further 30 min. Upon completion, the reaction mixture was quenched with sat. aq. NH4Cl (100 mL) at 25° C., diluted with water (100 mL) and extracted with EtOAc (3×150 mL). The combined organic layers were washed with brine (50 mL), dried over Na2SO4, evaporated and purified by column chromatography (PE/EtOAc=1/2) to give methyl 1-(1-(tert-butoxycarbonyl)-3-(nitromethyl)azetidin-3-yl)-1H-imidazole-2-carboxylate (0.72 g, 2.12 mmol, 11% yield) as a yellow solid.
m/z ES+ [M+H]+ 341.1; 1H NMR (400 MHz, CDCl3) δ ppm 7.23 (s, 2H), 5.25 (s, 2H), 4.56-4.30 (m, 4H), 3.97 (s, 3H), 1.46 (s, 9H).
Step d. This step was conducted in a similar manner to Intermediate A10, step c.
Step e. To a solution of tert-butyl 8′-oxo-7′,8′-dihydro-6′H-spiro[azetidine-3,5′-imidazo[1,2-a]pyrazine]-1-carboxylate (200 mg, 0.72 mmol mmol) in THF (3 mL) was added a solution of borane dimethyl sulfide complex (10 M in THF, 0.36 mL). The reaction mixture was stirred at 25° C. for 12 h. Upon completion, the reaction mixture was quenched with MeOH (2 mL) at 0° C., evaporated and purified by reverse phase flash chromatography (water (0.1% FA)/MeCN) to give the title compound (90 mg, 47% yield) as a yellow solid.
m/z ES+ [M+H]+ 265.1.
Steps a-b. These 2 steps were conducted as described in Intermediate A12, steps a-b.
Step c. This step was conducted in a similar manner to Intermediate A28, step c, using 2-bromo-1H-imidazole (CAS: 16681-56-4).
Step d. To a solution of tert-butyl 3-(2-bromo-1H-imidazol-1-yl)-3-(nitromethyl)azetidine-1-carboxylate (700 mg, 1.94 mmol) in MeOH (20 mL) was added cobalt (II) chloride (1.51 g, 11.6 mmol) and NaBH4 (440 mg, 11.6 mmol). The reaction mixture was stirred at 0° C. for 1 h. Upon completion, the mixture was quenched with sat. aq. NH4Cl (1 mL), filtered and the filtrate was evaporated. The residue was purified by reverse phase flash chromatography (water (0.1% FA)/MeCN) to give the title compound (300 mg, 61.9% yield) as a white solid.
m/z ES+ [M+H]+ 250.9.
Step a. To a suspension of NaH (128 mg, 3.20 mmol, 60% dispersion in mineral oil) in DMF (5 mL) was added benzyl 3-oxopiperazine-1-carboxylate (CAS: 78818-15-2; 0.5 g, 2.13 mmol) at 0° C. The mixture was stirred at 0° C. for 30 min under a N2 atmosphere, after which tert-butyl 3-iodoazetidine-1-carboxylate (CAS: 254454-54-1; 724 mg, 2.56 mmol) was added. The reaction mixture was stirred at 20° C. for a further 1 h. Upon completion, the reaction mixture was partitioned between water (40 mL) and EtOAc (40 mL). The aqueous layer was further extracted with EtOAc (2×40 mL) and the combined organic layers were dried over Na2SO4 and evaporated. The residue was purified by reverse phase flash chromatography (water (0.1% FA)/MeCN) to give benzyl 4-(1-(tert-butoxycarbonyl)azetidin-3-yl)-3-oxopiperazine-1-carboxylate (60 mg, 7% yield) as a yellow oil.
m/z ES+ [M+H]+ 390.1
Step b. To a solution of benzyl 4-(1-(tert-butoxycarbonyl)azetidin-3-yl)-3-oxopiperazine-1-carboxylate (60 mg, 0.15 mmol) in 2-propanol (2 mL) was added 10% Pd/C (5 mg). The reaction mixture was stirred at 40° C. for 12 h under a H2 (15 psi) atmosphere. Upon completion, the reaction mixture was filtered and the filtrate was evaporated to give the title compound (44 mg, crude) as a yellow oil which was used without further purification.
Step a. A solution of tert-butyl 7-oxa-3-azabicyclo[4.1.0]heptane-3-carboxylate (CAS: 161157-50-2; 1.0 g, 5.02 mmol) in methanamine (2M in THF, 50.2 mL) was stirred at 100° C. for 12 h. Upon completion, the reaction mixture was filtered and evaporated to give a yellow oil (1.15 g, crude). The oil was dissolved in THF (12 mL) and water (6 mL) and CbzCl (1.22 g, 7.16 mmol) and Na2CO3 (1.52 g, 14.3 mmol) were added. The reaction mixture was stirred at 25° C. for 1 h. Upon completion, the mixture was diluted with water (30 mL) and extracted with EtOAc (3×50 mL). The combined organic layers were washed with brine (3×20 mL), dried over Na2SO4 and evaporated. The residue was purified by column chromatography (PE/EtOAc=1/1) to give tert-butyl (rel-3R,4R)-3-(((benzyloxy)carbonyl)(methyl)amino)-4-hydroxypiperidine-1-carboxylate (200 mg, 11% yield) as a colourless oil. The regiochemistry was confirmed by 2D NMR.
m/z ES+ [M+H]+ 365.2; 1H NMR (400 MHz, CDCl3) δ ppm 7.44-7.29 (m, 5H), 5.22-5.10 (m, 2H), 4.23-3.98 (m, 2H), 3.82-3.74 (m, 2H), 2.93 (s, 3H), 2.89-2.74 (m, 1H), 2.70-2.64 (m, 1H), 2.05-1.99 (m, 1H), 1.59-1.50 (m, 1H), 1.46 (s, 9H).
Step b. To a solution of tert-butyl (rel-3R,4R)-3-(((benzyloxy)carbonyl)(methyl)amino)-4-hydroxypiperidine-1-carboxylate (200 mg, 0.55 mmol) in THF (5 mL) was added 10% Pd/C (30 mg). The mixture was stirred at 25° C. for 30 min under a H2 atmosphere (15 psi). Upon completion, the reaction mixture was filtered and the filtrate was evaporated to give the title compound (110 mg, crude) as a colourless oil which was used without further purification.
Step a. Ethyl 2,3-dibromopropanoate (CAS: 3674-13-3; 21.6 g, 83.2 mmol) was added dropwise to a solution of N1,N2-dibenzylethane-1,2-diamine (CAS: 140-28-3; 20 g, 83.2 mmol) and triethylamine (26 mL, 164 mmol) in toluene (300 mL) at 40° C. The reaction mixture was stirred at 80° C. for 4 h. Upon completion, the reaction mixture was cooled to rt and the solvent was evaporated under vacuum. The reaction mixture was diluted with EtOAc (300 mL) and washed with water (2×100 mL). The organic layer was further washed with brine, dried over dried over Na2SO4 and evaporated. Purification by column chromatography (n-hexane/EtOAc=4/1) gave ethyl 1,4-dibenzylpiperazine-2-carboxylate (20 g, 73% yield) as a pale yellow oil.
m/z ES+ [M+H]+ 339.7.
Step b. To a solution of ethyl 1,4-dibenzylpiperazine-2-carboxylate (10 g, 29.6 mmol) in THF (200 mL), cooled to −78° C., was added paraformaldehyde (0.89 g) followed by a solution of LiHMDS (1 M in THF, 88.8 mL, 88.8 mmol) under a N2 atmosphere. The reaction mixture was stirred at −78° C. for 15 min and then stirred at rt 2 h. Upon completion, the reaction mixture was quenched with water (200 mL) and the aqueous mixture was extracted with EtOAc (2×500 mL). The organic layers were combined, dried over dried over Na2SO4 and evaporated.
Purification by column chromatography (n-hexane/EtOAc=3/2) gave 5,8-dibenzyl-2,5,8-triazaspiro[3.5]nonan-1-one (5.6 g, 60% yield) as a white solid.
m/z ES+ [M+H]+ 322.2.
Step c. To a solution of 5,8-dibenzyl-2,5,8-triazaspiro[3.5]nonan-1-one (5.6 g, 17.4 mmol) in THF (60 mL), cooled to 0° C., was added a solution of LiAlH4 (2.5 M in THF, 20.9 mL, 52.3 mmol). The reaction mixture was stirred at 60° C. for 15 h. Upon completion, the reaction mixture was cooled to 0° C. and quenched with the slow addition of sat. aq. NH4Cl. The reaction mixture was extracted with EtOAc (2×100 mL). The organic layers were combined, dried over dried over Na2SO4 and evaporated to give 5,8-dibenzyl-2,5,8-triazaspiro[3.5]nonane (5.2 g, 97% crude) as a yellow solid.
m/z ES+ [M+H]+ 308.2.
Step d. To a solution of 5,8-dibenzyl-2,5,8-triazaspiro[3.5]nonane (5.2 g, 16.9 mmol) in methanol (52 mL) were added triethylamine (4.7 mL, 33.8 mmol), DMAP (0.12 g, 1.0 mmol) and di-tert-butyl decarbonate (4.6 g, 21.1 mmol). The reaction mixture was stirred at rt for 3 h. Upon completion, the reaction mixture was quenched with water (50 mL) and extracted with EtOAc (2×100 mL). The organic layers were combined, dried over Na2SO4 and evaporated. Purification by column chromatography (n-hexane/EtOAc=4/1) gave tert-butyl 5,8-dibenzyl-2,5,8-triazaspiro[3.5]nonane-2-carboxylate (5.1 g, 74% yield) as a yellow solid.
m/z ES+ [M+H]+ 408.3.
Step e. To a solution of tert-butyl 5,8-dibenzyl-2,5,8-triazaspiro[3.5]nonane-2-carboxylate (5.0 g, 12.3 mmol) in 1,2-dimethoxyethane (50 mL), cooled to 0° C., was added 1-chloroethyl chloroformate (0.69 g, 18.4 mmol). The reaction mixture was stirred at 80° C. for 2 h. Upon completion, the reaction mixture was poured into cold water (50 mL) and extracted with DCM (2×100 mL). The organic layers were combined, washed with cold water and brine, dried over Na2SO4 and evaporated. The crude was triturated with diethyl ether to give tert-butyl 5-benzyl-2,5,8-triazaspiro[3.5]nonane-2-carboxylate (1.5 g, 39% yield) as white solid.
m/z ES+ [M+H]+ 318.7.
Step f. To a solution of tert-butyl 5-benzyl-2,5,8-triazaspiro[3.5]nonane-2-carboxylate (2.5 g, 7.88 mmol) in MeOH (25 mL) was added 10% Pd/C (250 mg) under a N2 atmosphere. The suspension was degassed and purged with H2 3 times. The mixture was stirred at 25° C. for 4 h under a H2 atmosphere (15 psi). Upon completion, the reaction mixture was filtered and the filtrate was evaporated to give the title compound (1.2 g, crude) as yellow solid.
m/z ES+ [M+H]+ 228.2; 1H NMR (400 MHz, CDCl3) δ ppm 4.11-4.05 (m, 2H), 3.82-3.75 (m, 2H), 3.29 (s, 2H), 3.23-3.15 (m, 4H), 1.44 (s, 9H).
Step a. N-benzyl-1-methoxy-N-((trimethylsilyl)methyl)methanamine (CAS: 93102-05-7; 4.40 g, 18.5 mmol) and LiF (1.20 g, 46.3 mmol) were added to a solution of tert-butyl 3-(cyanomethylene)azetidine-1-carboxylate (CAS: 1153949-11-1; 3 g, 15.4 mmol) in MeCN (30 mL). The mixture was stirred at 60° C. for 72 h. Upon completion, the mixture was filtered and the filtrate was evaporated. The residue was purified by column chromatography (PE/EtOAc=3/1) to give tert-butyl 6-benzyl-8-cyano-2,6-diazaspiro[3.4]octane-2-carboxylate (3.7 g, 73% yield) as a colourless oil.
m/z ES+ [M+H]+ 328.1.
Step b. A mixture of tert-butyl 6-benzyl-8-cyano-2,6-diazaspiro[3.4]octane-2-carboxylate (3.2 g, 9.77 mmol), 10% Pd/C (1.0 g) and ammonium formate (3.08 g, 48.8 mmol) in EtOH (30 mL) was stirred at 80° C. for 12 h. Upon completion, the mixture was filtered and evaporated to give the title compound (2 g, crude) as a light yellow oil.
m/z ES+ [M+H]+ 238.2.
Step a. To a solution of diethyl (1-cyanoethyl)phosphonate (1.0 g, 5.23 mmol) in THF (50 mL) was added NaH (313 mg, 7.85 mmol, 60% dispersion in mineral oil) at 0° C. The mixture was stirred for 30 min, after which tert-butyl 3-oxoazetidine-1-carboxylate (1.79 g, 10.46 mmol) was added and the reaction mixture was slowly warmed to 15° C. and stirred for 12 h. Upon completion, the reaction was quenched with water (50 mL) and extracted with EtOAc (2×50 mL). The combined organic layers were evaporated and the residue was purified by column chromatography (PE/EtOAc=5/1) to give tert-butyl 3-(1-cyanoethylidene)azetidine-1-carboxylate (600 mg, 55% yield) as a colourless oil.
1H NMR (400 MHz, CDCl3) δ ppm 4.60-4.53 (m, 2H), 4.45-4.46 (m, 2H), 1.74 (t, J=1.6 Hz, 3H), 1.39 (s, 9H).
Steps b-c. These 2 steps were conducted in a similar manner to Intermediate A33, steps a-b.
m/z ES+ [M+H]+ 252.2; 1H NMR (400 MHz, CD3OD) δ ppm 4.06-3.93 (m, 2H), 3.81 (d, J=9.2 Hz, 1H), 3.68 (d, J=9.2 Hz, 1H), 3.26-3.17 (m, 2H), 3.16-3.08 (m, 1H), 2.86 (d, J=11.6 Hz, 1H), 1.40-1.35 (m, 12H).
Step a. A mixture of tert-butyl 3-iodoazetidine-1-carboxylate (CAS: 254454-54-1; 10.0 g, 35.3 mmol), methyl 1H-imidazole-4-carboxylate (CAS: 17325-26-7; 8.91 g, 70.6 mmol) and Cs2CO3 (34.5 g, 106 mmol) in DMF (100 mL) was stirred at 100° C. for 1 h. Upon completion, the reaction mixture was diluted with water (300 mL) and extracted with EtOAc (300 mL). The organic layer was washed with brine (300 mL), dried over Na2SO4 and evaporated. The residue was purified by reverse phase flash chromatography (water (0.1% NH4OH)/MeCN) to give methyl 1-(1-(tert-butoxycarbonyl)azetidin-3-yl)-1H-imidazole-4-carboxylate (5.5 g, 55% yield) as a white solid. The regiochemistry was confirmed by 2D NMR.
m/z ES+ [M+H]+ 282.2; 1H NMR (400 MHz, CDCl3) δ ppm 7.85 (d, J=1.6 Hz, 1H), 7.62 (d, J=1.6 Hz, 1H), 4.96-4.89 (m, 1H), 4.50-4.42 (m, 2H), 4.10-4.06 (m, 2H), 3.88 (s, 3H), 1.45 (s, 9H).
Step b. To a solution of methyl 1-(1-(tert-butoxycarbonyl)azetidin-3-yl)-1H-imidazole-4-carboxylate (2.00 g, 7.11 mmol) in THF (20 mL) was added LiBH4 (465 mg, 21.3 mmol) at 0° C. The reaction mixture was stirred at 0° C. for 1 h, and then MeOH (2 mL) was added. The reaction mixture was stirred at 25° C. for a further 12 h. Upon completion, the reaction mixture was poured into sat. aq. NH4Cl (50 mL) and extracted with EtOAc (50 mL). The organic layer was washed with brine (50 mL), dried over Na2SO4 and evaporated. The residue was purified by column chromatography (DCM/MeOH=10/1) to give tert-butyl 3-(4-(hydroxymethyl)-1H-imidazol-1-yl)azetidine-1-carboxylate (0.65 g, 36% yield) as a white solid.
1H NMR (400 MHz, CDCl3) δ ppm 7.56 (d, J=1.2 Hz, 1H), 7.12 (s, 1H), 4.92-4.81 (m, 1H), 4.61 (s, 2H), 4.46-4.37 (m, 2H), 4.14-4.04 (m, 2H), 1.46 (s, 9H).
Step c. To a solution of tert-butyl 3-(4-(hydroxymethyl)-1H-imidazol-1-yl)azetidine-1-carboxylate (50 mg, 0.20 mmol) in DCM (1 mL) was added MnO2 (52 mg, 0.59 mmol). The reaction mixture was stirred at 25° C. for 2 h. Upon completion, the reaction mixture was filtered and evaporated to give the title compound (40 mg, 81% yield) as a brown oil.
1H NMR (400 MHz, CDCl3) δ ppm 9.91 (s, 1H), 7.88 (s, 1H), 7.72 (s, 1H), 5.05-4.86 (m, 1H), 4.60-4.40 (m, 2H), 4.20-4.04 (m, 2H), 1.49 (s, 9H).
The title compound was prepared in a similar manner to Intermediate B1, using methyl 1H-pyrazole-3-carboxylate (CAS: 15366-34-4) in step a.
m/z ES+ [M+Na]+ 274.1.
Step a. This step was conducted in a similar manner to Intermediate B1, step a, using methyl 1H-1,2,4-triazole-3-carboxylate (CAS: 4928-88-5).
Step b. To a solution of methyl 1-(1-(tert-butoxycarbonyl)azetidin-3-yl)-1H-1,2,4-triazole-3-carboxylate (250 mg, 0.89 mmol) in DCM (10 mL) was added DIBAL-H (1 M in DCM, 2.66 mL). The reaction mixture was stirred at −70° C. for 1 h. Upon completion, the mixture was quenched with MeOH (0.5 mL) and stirred at 20° C. for 20 min. The mixture was filtered and evaporated to give the title compound (170 mg, 76% yield) as a white solid.
1H NMR (400 MHz, CDCl3) δ ppm 10.08 (s, 1H), 8.32 (s, 1H), 5.28-5.17 (m, 1H), 4.61-4.20 (m, 4H), 1.51 (s, 9H).
Step a. To a solution of tert-butyl 3-hydroxyazetidine-1-carboxylate (CAS: 141699-55-0; 10 g, 57.7 mmol) in THF (100 mL) was added NaH (346 mg, 8.66 mmol, 60% dispersion in mineral oil). The mixture was stirred at 20° C. for 30 min. Then methyl acrylate (CAS: 96-33-3; 24.9 g, 289 mmol) was added and the reaction mixture was stirred at 20° C. for 12 h. Upon completion, the mixture was quenched with sat. aq. NH4Cl (100 mL) and extracted with EtOAc (2×100 mL). The combined organic layers were washed with brine (200 mL), dried over Na2SO4, and evaporated. The crude product was purified by column chromatography (PE/EtOAc=5/1) to give tert-butyl 3-(3-methoxy-3-oxopropoxy)azetidine-1-carboxylate (6 g, 40% yield) as a colourless oil.
1H NMR (400 MHz, CDCl3) δ ppm 4.27-4.19 (m, 1H), 4.09-4.01 (m, 2H), 3.82 (dd, J=4.4, 9.6 Hz, 2H), 3.71 (s, 3H), 3.64 (t, J=6.4 Hz, 2H), 2.59 (t, J=6.4 Hz, 2H), 1.43 (s, 9H).
Step b. To a solution of tert-butyl 3-(3-methoxy-3-oxopropoxy)azetidine-1-carboxylate (2 g, 7.71 mmol) in DCM (50 mL) was added DIBAL-H (1 M in THF, 23.1 mL) dropwise at −70° C. Then the reaction mixture was stirred at −70° C. for 30 min. Upon completion, the mixture was quenched with MeOH (5 mL) at −70° C., and allowed to warm to 20° C. over 30 min. The mixture was filtered and evaporated to give the title compound (1.4 g, 79% yield) as a colourless oil, which was used for the next step directly without further purification.
1H NMR (400 MHz, CDCl3) δ ppm 9.80 (t, J=1.6 Hz, 1H), 4.26-4.20 (m, 1H), 4.12-4.04 (m, 2H), 3.84-3.78 (m, 2H), 3.74-3.66 (m, 2H), 2.76-2.64 (m, 2H), 1.44 (s, 9H).
Step a. This step was conducted in a similar manner to Intermediate B1, step a, using methyl 1H-pyrazole-4-carboxylate (CAS: 51105-90-9).
Step b. This step was conducted in a similar manner to Intermediate B3, step b.
Step c. This step was conducted in a similar manner to Intermediate B1, step c.
1H NMR (400 MHz, CDCl3) δ ppm 9.91 (s, 1H), 8.08 (s, 1H), 8.07 (s, 1H), 5.13-5.07 (m, 1H), 4.47-4.42 (m, 2H), 4.37-4.32 (m, 2H), 1.49 (s, 9H).
Step a. To a solution of tert-butyl 3-formylazetidine-1-carboxylate (4.8 g, 25.9 mmol) and glyoxal (40 wt % in water; 18.0 g, 129 mmol) in MeOH (50 mL) was added 30% ammonium hydroxide (18.1 g, 155 mmol). The reaction mixture was stirred at 20° C. for 12 h. Upon completion, the mixture was evaporated and the residue was purified by reverse phase flash chromatography (water (0.1% NH4OH)/MeCN) to give tert-butyl 3-(1H-imidazol-2-yl)azetidine-1-carboxylate (5.79 g, crude) as a brown oil.
m/z ES+ [M+H]+ 224.2.
Step b. To a solution of tert-butyl 3-(1H-imidazol-2-yl)azetidine-1-carboxylate (5.79 g, 25.9 mmol) in THF (3 mL) was added NaH (1.24 g, 31.1 mmol, 60% dispersion in mineral oil). The mixture was stirred at 0° C. for 30 min, after which SEMCl (5.62 g, 33.7 mmol) was added and the reaction mixture was stirred for a further 1 h. Upon completion, the mixture was quenched with sat. aq. NH4Cl (5 mL), diluted with water (100 mL) and extracted with EtOAc (3×100 mL). The combined organic layers were evaporated and the residue was purified by reverse phase flash chromatography (water (0.1% NH4OH)/MeCN) to give tert-butyl 3-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-imidazol-2-yl)azetidine-1-carboxylate (6.0 g, 65% yield) as a brown oil.
m/z ES+ [M+H]+ 354.0.
Step c. A mixture of tert-butyl 3-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-imidazol-2-yl)azetidine-1-carboxylate (3.1 g, 8.77 mmol), NBS (1.56 g, 8.77 mmol) in DCM (30 mL) and DMF (30 mL) was stirred at 25° C. for 12 h under a N2 atmosphere. Upon completion, the mixture was filtered and the filtrate was evaporated. The residue was purified by reverse phase flash chromatography (water (0.1% NH4OH)/MeCN) to give tert-butyl 3-(4-bromo-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-imidazol-2-yl)azetidine-1-carboxylate (3.2 g, 84% yield) as a yellow oil.
m/z ES+ [M+H]+ 434.2; 1H NMR (400 MHz, CDCl3) δ ppm 7.03-6.93 (m, 1H), 5.21-5.10 (m, 2H), 4.29-4.17 (m, 4H), 3.99-3.85 (m, 1H), 3.55-3.42 (m, 2H), 1.45 (s, 9H), 0.89 (t, J=8.4 Hz, 2H), 0.00 (s, 9H).
Step d. A mixture of tert-butyl 3-(4-bromo-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-imidazol-2-yl)azetidine-1-carboxylate (3.2 g, 7.40 mmol), Pd(dppf)Cl2 (270 mg, 0.37 mmol), TEA (793 mg, 7.84 mmol) in MeOH (30 mL) was stirred at 60° C. for 12 h under a CO atmosphere (50 psi). Upon completion, the mixture was filtered and the filtrate was evaporated. The residue was purified by by reverse phase flash chromatography (water (0.1% NH4OH)/MeCN) to give the title compound (3.0 g, 69% yield) as a yellow oil.
m/z ES+ [M+H]+ 412.2.
Step e. This step was conducted in a similar manner to Intermediate B3, step b.
Step f. This step was conducted in a similar manner to Intermediate B1, step c.
m/z ES+ [M+H]+ 382.2.
Step a. To a solution of ethyl 2-chlorooxazole-4-carboxylate (CAS: 460081-18-9; 1.0 g, 5.70 mmol) in DMF (10 mL) was added K2CO3 (1.57 g, 11.3 mmol) and tert-butyl piperazine-1-carboxylate (1.27 g, 6.83 mmol). The mixture was stirred at 25° C. for 5 h. Upon completion, the reaction mixture was diluted with water (20 mL) and extracted with EtOAc (3×10 mL). The combined organic layers were evaporated and the residue was purified by column chromatography (PE/EtOAc=3/1) to give ethyl 2-(4-(tert-butoxycarbonyl)piperazin-1-yl)oxazole-4-carboxylate (1.3 g, 68% yield) as a light yellow solid.
m/z ES+ [M+H]+ 326.2; 1H NMR (400 MHz, CDCl3) δ ppm 7.79 (s, 1H), 4.41-4.32 (m, 2H), 3.55-3.49 (m, 8H), 1.48 (s, 9H), 1.38-1.33 (m, 3H).
Step b. This step was conducted in a similar manner to Intermediate B3, step b.
m/z ES+ [M+H]+ 282.1.
Step a. To a mixture of N,N-dimethyl-1H-imidazole-1-sulfonamide (CAS: 78162-58-0; 9 g, 51.3 mmol) in THF (90 mL) was added n-BuLi (2.5 M in THF, 24.6 mL) dropwise at −70° C. The mixture was stirred at −70° C. for 30 min, after which CBr4 (18.7 g, 56.5 mmol) was added at −70° C. The reaction mixture was stirred at 20° C. for 16 h. Upon completion, the reaction mixture was quenched with sat. aq. NH4Cl (300 mL) and extracted with EtOAc (3×200 mL). The combined organic layers were washed with brine (3×500 mL) and evaporated. The residue was purified by column chromatography (PE/EtOAc=2/1) to give 2-bromo-N,N-dimethyl-1H-imidazole-1-sulfonamide (10.0 g, 77% yield) as a red solid.
m/z ES+ [M+H]+ 255.9; 1H NMR (400 MHz, CDCl3) δ ppm 7.42 (d, J=1.6 Hz, 1H), 7.01 (d, J=1.6 Hz, 1H), 3.02 (s, 6H).
Step b. A mixture of 2-bromo-N,N-dimethyl-1H-imidazole-1-sulfonamide (2 g, 7.87 mmol) and tert-butyl piperazine-1-carboxylate (CAS: 143238-38-4; 14.6 g, 78.7 mmol) was stirred at 90° C. for 20 h. Upon completion, the reaction mixture was evaporated and the residue was purified by reverse phase flash chromatography (water (0.1% FA)/MeCN) to give tert-butyl 4-(1-(N,N-dimethylsulfamoyl)-1H-imidazol-2-yl)piperazine-1-carboxylate (1 g, 35% yield) as a yellow solid.
m/z ES+ [M+H]+ 359.9; 1H NMR (400 MHz, CDCl3) δ ppm 7.09 (d, J=2.0 Hz, 1H), 6.78 (d, J=2.0 Hz, 1H), 3.59-3.52 (m, 4H), 3.23-3.18 (m, 4H), 2.89 (s, 6H), 1.48 (s, 9H).
Step c. To a mixture of tert-butyl 4-(1-(N,N-dimethylsulfamoyl)-1H-imidazol-2-yl)piperazine-1-carboxylate (647 mg, 1.80 mmol) in THF (6 mL) was added n-BuLi (2.5 M in THF, 1.44 mL) dropwise at −70° C. The mixture was stirred at −70° C. for 30 min, after which DMF (789 mg, 10.8 mmol) was added at −70° C. The reaction mixture was stirred at 20° C. for 2 h. Upon completion, the reaction mixture was quenched with sat. aq. Na2CO3 (30 mL) and extracted with EtOAc (3×10 mL). The combined organic layers were evaporated and the residue was purified by reverse phase flash chromatography (water (0.1% FA)/MeCN) followed by Prep-TLC (EtOAc) to give tert-butyl 4-(1-(N,N-dimethylsulfamoyl)-5-formyl-1H-imidazol-2-yl)piperazine-1-carboxylate (154 mg, 22% yield) as a yellow solid.
1H NMR (400 MHz, CDCl3) δ ppm 9.83 (s, 1H), 7.63 (s, 1H), 3.62-3.57 (m, 4H), 3.47-3.42 (m, 4H), 2.84 (s, 6H), 1.49 (s, 9H).
Step d. A mixture of tert-butyl 4-(1-(N,N-dimethylsulfamoyl)-5-formyl-1H-imidazol-2-yl)piperazine-1-carboxylate (160 mg, 0.41 mmol) in conc. HCl (2 mL) was stirred at 60° C. for 2 h. Upon completion, the reaction mixture was basified to pH 7 with sat aq. Na2CO3 and extracted with EtOAc (3×10 mL). The combined organic layers were evaporated to give 2-(piperazin-1-yl)-1H-imidazole-4-carbaldehyde (70 mg, 94% yield) as a yellow solid.
m/z ES+ [M+H]+ 181.2.
Step e. To a mixture of 2-(piperazin-1-yl)-1H-imidazole-4-carbaldehyde (70 mg, 0.39 mmol) and Na2CO3 (82 mg, 0.78 mmol) in THF (3 mL) was added Boc2O (169 mg, 0.78 mmol). The reaction mixture was stirred at 20° C. for 30 min. Upon completion, the reaction mixture was filtered and the filtrate was evaporated. The residue was purified by reverse phase flash chromatography (water (0.1% FA)/MeCN) to give the title compound (80 mg, 73% yield) as a white solid.
m/z ES+ [M+H]+ 281.3; 1H NMR (400 MHz, CDCl3) δ ppm 9.83 (s, 1H), 9.31 (s, 1H), 7.61 (s, 1H), 3.57 (s, 8H), 1.49 (s, 9H).
The title compound was prepared in a similar manner to Intermediate B3, using methyl 1H-1,2,4-triazole-3-carboxylate (CAS: 4928-88-5) and tert-butyl 4-iodopiperidine-1-carboxylate (CAS: 301673-14-3) in step a.
1H NMR (400 MHz, CDCl3) δ ppm 10.01 s, 1H), 8.24 (s, 1H), 4.49-4.45 (m, 1H), 4.32-4.28 (m, 2H), 2.97-2.91 (m, 2H), 2.22-2.17 (m, 2H), 2.08-1.98 (m, 2H), 1.49 (s, 9H).
Step a. A mixture of methyl 3-bromobenzoate (500 mg, 2.33 mmol), tert-butyl 3-bromoazetidine-1-carboxylate (CAS: 1064194-10-0; 714 mg, 3.02 mmol), (Ir[dF(CF3)ppy]2(dtbpy))PF6 (CAS: 870987-63-6; 26 mg, 0.023 mmol), [4,4′-bis(1,1-dimethylethyl)-2,2′-bipyridine]nickel (II) dichloride (CAS: 1034901-50-2; 4.6 mg, 0.012 mmol), tris(trimethylsilyl)silane (CAS: 1873-77-4; 578 mg, 2.33 mmol) and Na2CO3 (493 mg, 4.65 mmol) in 1,2-dimethoxyethane (50 mL) was degassed and purged with N2 3 times and then the mixture was stirred at 25° C. for 14 h under a N2 atmosphere. Upon completion, the reaction mixture was filtered and evaporated. The residue was diluted with water (20 mL) and extracted with EtOAc (3×30 mL). The combined organic layers were dried over Na2SO4, evaporated and purified by column chromatography (PE/EtOAc=9/1) to give tert-butyl 3-(3-(methoxycarbonyl)phenyl)azetidine-1-carboxylate (590 mg, 75% yield) as a light yellow oil.
1H NMR (400 MHz, CDCl3) δ ppm 8.00-7.98 (m, 1H), 7.95-7.92 (m, 1H), 7.54-7.49 (m, 1H), 7.46-7.41 (m, 1H), 4.35 (t, J=8.8 Hz, 2H), 4.00-3.97 (m, 2H), 3.93 (s, 3H), 3.83-3.75 (m, 1H), 1.47 (s, 9H).
Step b. A mixture of tert-butyl 3-(3-(methoxycarbonyl)phenyl)azetidine-1-carboxylate (100 mg, 0.34 mmol) in DCM (5 mL) was degassed and purged with N2 3 times. DIBAL-H (1 M in toluene, 1.03 mL) was added and the reaction mixture was stirred at −60° C. for 1 h under a N2 atmosphere. Upon completion, the reaction mixture was quenched with MeOH (0.5 mL) at −60° C., filtered and evaporated to give tert-butyl 3-(3-(hydroxymethyl)phenyl)azetidine-1-carboxylate (90 mg, crude) as a light yellow oil.
1H NMR (400 MHz, CDCl3) δ ppm 7.37-7.33 (m, 2H), 7.27-7.23 (m, 2H), 4.73 (s, 2H), 4.33-4.30 (m, 2H), 4.02-3.95 (m, 2H), 3.84-3.68 (m, 1H), 1.48 (s, 9H).
Step c. To a solution of tert-butyl 3-(3-(hydroxymethyl)phenyl)azetidine-1-carboxylate (90 mg, 0.34 mmol) in DCM (1 mL) was added MnO2 (594 mg, 6.84 mmol). The mixture was stirred at 25° C. for 16 h. Upon completion, the reaction mixture was filtered and evaporated to give the title compound (70 mg, crude) as a light yellow oil.
1H NMR (400 MHz, CDCl3) δ ppm 10.04 (s, 1H), 7.87 (s, 1H), 7.85-7.78 (m, 1H), 7.65-7.52 (m, 2H), 4.42-4.31 (m, 2H), 4.03-3.96 (m, 2H), 3.87-3.78 (m, 1H), 1.48 (s, 9H).
Step a. This step was conducted in a similar manner to Intermediate B10, step a, using methyl 6-bromopicolinate (CAS: 26218-75-7).
Step b. To a solution of methyl 6-(1-(tert-butoxycarbonyl)azetidin-3-yl)picolinate (100 mg, 0.34 mmol) in DCM (5 mL) was added DIBAL-H (1 M in THF, 1.03 mL) at −70° C. The reaction mixture was stirred at −70° C. for 1 h. Upon completion, the mixture was quenched with MeOH (0.2 mL), stirred at 20° C. for 30 min, filtered and the filtrate was evaporated to give the title compound (80 mg, 89% yield) as a white solid.
1H NMR (400 MHz, CDCl3) δ ppm 10.09 (s, 1H), 7.89-7.80 (m, 2H), 7.50-7.43 (m, 1H), 4.38-4.33 (m, 2H), 4.26-4.20 (m, 2H), 4.04-3.91 (m, 1H), 1.48 (s, 9H).
The title compound was prepared in a similar manner to Intermediate B10, using methyl 2-bromopyrimidine-4-carboxylate (CAS: 1209459-78-8) in step a.
1H NMR (400 MHz, CDCl3) δ ppm 10.05 (s, 1H), 9.01 (d, J=4.8 Hz, 1H), 7.70 (d, J=4.8 Hz, 1H), 4.46-4.26 (m, 4H), 4.18-4.10 (m, 1H), 1.48 (s, 9H).
Step a. This step was conducted in a similar manner to Intermediate B10, step a, using tert-butyl 3-bromoazetidine-1-carboxylate and methyl 4-bromobenzoate.
Steps b-c. These 2 steps were conducted in a similar manner to Intermediate B1, steps b-c.
m/z ES+ [M-tBu+H]+ 206.1; 1H NMR (400 MHz, CDCl3) δ ppm 10.01 (s, 1H), 7.88 (d, J=8.0 Hz, 2H), 7.49 (d, J=8.0 Hz, 2H), 4.38 (t, J=8.8 Hz, 2H), 3.99 (t, J=8.8 Hz, 2H), 3.86-3.78 (m, 1H), 1.48 (s, 9H).
Step a. This step was conducted in a similar manner to Intermediate B10, step a, using tert-butyl 3-bromoazetidine-1-carboxylate and methyl 4-bromobenzoate.
Step b. This step was conducted in a similar manner to Intermediate A23a and Intermediate A23b, step b.
Step c. This step was conducted in a similar manner to Intermediate B3, step b.
m/z ES+ [M-tBu+H]+ 212.0; 1H NMR (400 MHz, CDCl3) δ ppm 9.72-9.59 (m, 1H), 3.98-3.89 (m, 2H), 3.69-3.56 (m, 2H), 2.48-1.93 (m, 5H), 1.90-1.75 (m, 2H), 1.64-1.49 (m, 2H), 1.43 (d, J=2.0 Hz, 9H), 1.33-1.11 (m, 1H), 1.01-0.81 (m, 1H).
The title compound was prepared in a similar manner to Intermediate B13, using tert-butyl 3-bromoazetidine-1-carboxylate and methyl 3-bromo-4-fluorobenzoate (CAS: 82702-31-6).
1H NMR (400 MHz, CDCl3) δ ppm 9.99 (s, 1H), 7.95 (dd, J=7.2, 2.0 Hz, 1H), 7.85-7.79 (m, 1H), 7.22 (dd, J=9.6, 8.8 Hz, 1H), 4.40-4.32 (m, 2H), 4.10-4.01 (m, 3H), 1.48 (s, 9H).
The title compound was prepared in a similar manner to Intermediate B10, using methyl 4-bromobenzoate and (tert-butoxycarbonyl)proline (CAS: 59433-50-0) in step a.
1H NMR (400 MHz, CDCl3) δ ppm 10.00 (s, 1H), 7.83 (d, J=8.0 Hz, 2H), 7.34 (d, J=8.0 Hz, 2H), 5.00-4.83 (m, 1H), 3.66-3.64 (m, 2H), 2.39-2.37 (m, 1H), 1.92-1.80 (m, 3H), 1.48-1.17 (m, 9H).
Step a. A mixture of tert-butyl 6-bromo-3,4-dihydroisoquinoline-2(1H)-carboxylate (CAS: 893566-74-0; 1.0 g, 3.20 mmol), N-formylsaccharin (CAS: 50978-45-5; 1.35 g, 6.41 mmol), Pd(OAc)2 (58 mg, 0.26 mmol), 1,4-bis(diphenylphosphino)butane (CAS: 7688-25-7; 163 mg, 0.38 mmol), triethylsilane (819 mg, 7.05 mmol) and Na2CO3 (848 mg, 8.01 mmol) in DMF (20 mL) was stirred at 80° C. for 16 h under a N2 atmosphere. Upon completion, the reaction mixture was filtered, evaporated and purified by column chromatography (PE/EtOAc=5/1) to give the title compound (100 mg, 11% yield) as a colourless oil.
1H NMR (400 MHz, CDCl3) δ ppm 9.90 (s, 1H), 7.69-7.54 (m, 2H), 7.21 (s, 1H), 4.58 (s, 2H), 3.61 (t, J=5.6 Hz, 2H), 2.85 (t, J=5.6 Hz, 2H), 1.43 (s, 9H).
The title compound was prepared in a similar manner to Intermediate B17, using tert-butyl 5-bromoisoindoline-2-carboxylate (CAS: 201940-08-1).
1H NMR (400 MHz, CDCl3) δ ppm 10.00 (s, 1H), 7.81-7.75 (m, 2H), 7.45-7.38 (m, 1H), 4.75-4.72 (m, 4H), 1.52 (s, 9H).
Step a. To a solution of 7-(tert-butyl) 2-ethyl 5,6-dihydroimidazo[1,2-a]pyrazine-2,7(8H)-dicarboxylate (CAS: 1053656-22-6; 300 mg, 1.07 mmol) in DCM (8 mL) was added DIBAL-H (1 M in toluene, 3.20 mL). The reaction mixture was stirred at −70° C. for 1 h. Upon completion, the mixture was quenched with MeOH (0.7 mL), stirred at 20° C. for 30 min, filtered and the filtrate was evaporated to give the title compound (210 mg, 78% yield) as a white solid.
1H NMR (400 MHz, CDCl3) δ ppm 9.84 (s, 1H), 7.56 (s, 1H), 4.74 (s, 2H), 4.09 (t, J=7.2 Hz, 2H), 3.89 (t, J=7.2 Hz, 2H), 1.49 (s, 9H).
Step a. To a solution of NaH (1.59 g, 39.6 mmol, 60% dispersion in mineral oil) in DMF (100 mL) was added methyl (tert-butoxycarbonyl)glycinate (CAS: 31954-27-5; 5 g, 26.4 mmol) at 0° C. The reaction mixture was stirred at 0° C. for 30 min, then 3-bromoprop-1-ene (3.20 g, 26.4 mmol) was added. The reaction mixture was stirred at 0° C. for a further 3 h. Upon completion, the reaction mixture was quenched with sat. aq. NH4Cl (20 mL) at 0° C. and then diluted with water (100 mL) and extracted with EtOAc (3×50 mL). The combined organic layers were washed with brine (50 mL), dried over Na2SO4, evaporated and purified by column chromatography (PE/EtOAc=5/1) to give methyl N-allyl-N-(tert-butoxycarbonyl)glycinate (3.0 g, 50% yield) as a colourless oil.
1H NMR (400 MHz, CDCl3) δ ppm 5.76-5.65 (m, 1H), 5.11-5.03 (m, 2H), 3.93-3.73 (m, 4H), 3.66 (s, 3H), 1.40-1.32 (m, 9H).
Step b. A mixture of methyl N-allyl-N-(tert-butoxycarbonyl)glycinate (500 mg, 2.18 mmol), OsO4 (55.4 mg, 0.22 mmol), NaIO4 (1.40 g, 6.54 mmol) in THF (5 mL) and water (5 mL) was stirred at 0° C. for 3 h under a N2 atmosphere. Upon completion, the mixture was quenched with sat. aq. Na2S2O3 (5 mL), stirred at 25° C. for a further 30 min, then extracted with EtOAc (3×20 mL). The combined organic layers were washed with sat. aq. Na2S2O3 (2×15 mL) and brine (20 mL), dried over Na2SO4, filtered and evaporated to give methyl N-(tert-butoxycarbonyl)-N-(2-oxoethyl)glycinate (350 mg, 1.51 mmol, 69% yield) as a black oil.
1H NMR (400 MHz, CDCl3) δ ppm 9.69 (d, J=7.2 Hz, 1H), 4.15-3.91 (m, 4H), 3.77 (s, 3H), 1.47 (s, 9H).
Step c. A mixture of benzyl hydrazinecarboxylate (862 mg, 5.19 mmol), methyl N-(tert-butoxycarbonyl)-N-(2-oxoethyl)glycinate (1.2 g, 5.19 mmol), NaBH3CN (326 mg, 5.19 mmol), 4 Å molecular sieves (300 mg) and acetic acid (1.56 g, 25.9 mmol) in MeOH (30 mL) was stirred at 25° C. for 6 h under a N2 atmosphere. Upon completion, the reaction mixture was filtered and the filtrate was evaporated. The residue was purified by column chromatography (PE/EtOAc=1/1) to give benzyl 2-(2-((tert-butoxycarbonyl)(2-methoxy-2-oxoethyl)amino)ethyl)hydrazine-1-carboxylate (1.5 g, 76% yield) as a colourless oil.
1H NMR (400 MHz, CDCl3) δ ppm 7.33-7.23 (m, 5H), 7.20 (br s, 1H), 5.07 (s, 2H), 3.95-3.77 (m, 2H), 3.66 (s, 3H), 3.42-3.29 (m, 2H), 2.99-2.83 (m, 2H), 1.43-1.31 (m, 9H).
Step d. To a solution of benzyl 2-(2-((tert-butoxycarbonyl)(2-methoxy-2-oxoethyl)amino)ethyl)hydrazine-1-carboxylate (1.5 g, 3.93 mmol) in MeOH (30 mL) was added 10% Pd/C (150 mg) under a N2 atmosphere. The suspension was degassed and purged with H2 3 times. The mixture was stirred under a H2 atmosphere (15 psi) at 25° C. for 5 h. Upon completion, the reaction mixture was filtered and evaporated to give tert-butyl 4-amino-3-oxopiperazine-1-carboxylate (0.8 g, 95% yield) as a yellow solid.
1H NMR (400 MHz, CDCl3) δ ppm 4.16 (s, 2H), 3.76-3.68 (m, 2H), 3.58-3.54 (m, 2H), 1.49 (s, 9H).
Step e. A mixture of tert-butyl 4-amino-3-oxopiperazine-1-carboxylate (0.8 g, 3.72 mmol), ethyl 2-ethoxy-2-iminoacetate (CAS: 816-27-3; 540 mg, 3.72 mmol) in EtOH (10 mL) was degassed and purged with N2 3 times. The mixture was stirred at 25° C. for 1 h under a N2 atmosphere. Upon completion, the mixture was evaporated and purified by reverse phase flash chromatography (water (0.1% FA)/MeCN) to give 7-(tert-butyl) 2-ethyl 5,6-dihydro-[1,2,4]triazolo[1,5-a]pyrazine-2,7(8H)-dicarboxylate (500 mg, 45% yield) as a colourless oil.
m/z ES+ [M+H]+ 297.1; 1H NMR (400 MHz, CDCl3) δ ppm 4.81 (s, 2H), 4.47 (q, J=7.2 Hz, 2H), 4.31 (t, J=5.6 Hz, 2H), 3.96 (t, J=5.2 Hz, 2H), 1.50 (s, 9H), 1.43 (t, J=7.2 Hz, 3H).
Step f. To a solution of 7-(tert-butyl) 2-ethyl 5,6-dihydro-[1,2,4]triazolo[1,5-a]pyrazine-2,7(8H)-dicarboxylate (100 mg, 0.34 mmol) in THF (5 mL) was added DIBAL-H (1 M in toluene, 1.01 mL). The mixture was stirred at −78° C. for 2 h. Upon completion, the reaction mixture was quenched with MeOH (1 mL) at −78° C. and then stirred for a further 2 h. The mixture was filtered and evaporated to give the title compound (60 mg, 71% yield) as a white solid.
1H NMR (400 MHz, CDCl3) δ ppm 10.01 (s, 1H), 4.84 (s, 2H), 4.34 (t, J=5.4 Hz, 2H), 4.02 (t, J=5.4 Hz, 2H), 1.52 (s, 9H).
The title compound was prepared in a similar manner to Intermediate B19, using 7-(tert-butyl) 3-ethyl 5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazine-3,7(8H)-dicarboxylate (CAS: 1215852-11-1).
1H NMR (400 MHz, CDCl3) δ ppm 10.10 (s, 1H), 4.93 (s, 2H), 4.43-4.38 (m, 2H), 3.90-3.87 (m, 2H), 1.52 (s, 9H).
Steps a-b. These 2 steps were conducted in a similar manner to Intermediate A25, steps a-b, using ethyl piperidine-4-carboxylate (CAS: 1126-09-6) in step b.
Step c. A mixture of ethyl 1-(1-benzhydryl-3-methylazetidin-3-yl)piperidine-4-carboxylate (580 mg, 1.48 mmol), 10% Pd/C (10 mg), 10% Pd(OH)2/C (10 mg) and 4 M HCl in MeOH (0.37 mL, 1.48 mmol) in MeOH (5 mL) was stirred at 25° C. for 16 h. Upon completion, the mixture was filtered and the filtrate was evaporated to give ethyl 1-(3-methylazetidin-3-yl)piperidine-4-carboxylate (320 mg, 95% yield) as colourless oil. A mixture of ethyl 1-(3-methylazetidin-3-yl)piperidine-4-carboxylate (220 mg, 0.97 mmol), Boc2O (318 mg, 1.46 mmol) and TEA (295 mg, 2.92 mmol) in THF (2 mL) was stirred at 25° C. for 16 h. Upon completion, the reaction mixture was evaporated and the residue was purified by column chromatography (PE/EtOAc=2/1) to give ethyl 1-(1-(tert-butoxycarbonyl)-3-methylazetidin-3-yl)piperidine-4-carboxylate (300 mg, 94% yield) as white solid.
m/z ES+ [M+H]+ 327.1; 1H NMR (400 MHz, CDCl3) δ ppm 4.21-4.11 (m, 2H), 3.78 (d, J=8.0 Hz, 2H), 3.52 (d, J=7.6 Hz, 2H), 2.71-2.55 (m, 2H), 2.37-2.07 (m, 3H), 2.00-1.88 (m, 2H), 1.84-1.69 (m, 2H), 1.46 (s, 9H), 1.28-1.20 (m, 6H).
Step d. This step was conducted in a similar manner to Intermediate B3, step b.
1H NMR (400 MHz, CDCl3) δ ppm 9.67 (s, 1H), 3.77 (d, J=8.0 Hz, 2H), 3.58-3.48 (m, 2H), 2.70-2.53 (m, 2H), 2.32-2.16 (m, 3H), 2.01-1.87 (m, 2H), 1.76-1.62 (m, 2H), 1.46 (s, 9H), 1.26 (s, 3H).
Step a. A mixture of tert-butyl 3-(methylamino)azetidine-1-carboxylate (100 mg, 0.54 mmol), TEA (271 mg, 2.68 mmol) and methyl acrylate (138 mg, 1.61 mmol) in MeOH (1 mL) was stirred at 25° C. for 16 h. Upon completion, the reaction mixture was diluted with water (10 mL) and extracted with EtOAc (3×10 mL). The combined organic layers were washed with brine (10 mL), dried over Na2SO4 and evaporated. The residue was purified by column chromatography (EtOAc) to give tert-butyl 3-((3-methoxy-3-oxopropyl)(methyl)amino)azetidine-1-carboxylate (100 mg, 68% yield) as a yellow oil.
1H NMR (400 MHz, CDCl3) δ ppm 3.97-3.88 (m, 2H), 3.86-3.74 (m, 2H), 3.70 (s, 3H), 3.29-3.13 (m, 1H), 2.68-2.42 (m, 4H), 2.15 (s, 3H), 1.44 (s, 9H).
Step b. This step was conducted in a similar manner to Intermediate B3, step b.
1H NMR (400 MHz, CDCl3) δ ppm 9.81 (s, 1H), 3.98-3.91 (m, 2H), 3.85-3.75 (m, 2H), 3.26-3.12 (m, 1H), 2.68-2.60 (m, 4H), 2.21 (m, 3H), 1.46 (s, 9H).
Intermediate B24: tert-butyl (7S)-7-formyl-8-methyloctahydro-2H-pyrazino[1,2-a]pyrazine-2-carboxylate
Step a. To a solution of 1-(tert-butyl) 3-methyl piperazine-1,3-dicarboxylate (5.0 g, 20.4 mmol) and ((benzyloxy)carbonyl)-D-serine (4.90 g, 20.4 mmol) in DCM (100 mL) was added HATU (7.78 g, 20.4 mmol) and DIPEA (5.29 g, 40.9 mmol). The mixture was stirred at rt for 1 h. Upon completion, the reaction mixture was diluted with water (100 mL) and extracted with EtOAc (3×100 mL). The combined organic layers were evaporated and the residue was purified by reverse phase flash chromatography (water (0.1% FA)/MeCN) to give 1-(tert-butyl) 3-methyl 4-(((benzyloxy)carbonyl)-D-seryl)piperazine-1,3-dicarboxylate (4.3 g, 44% yield) as an off-white solid.
m/z ES+ [M+H]+ 466.1; 1H NMR (400 MHz, DMSO-d6) δ ppm 7.38-7.31 (m, 5H), 5.07-4.92 (m, 3H), 4.92-4.71 (m, 1H), 4.67-4.45 (m, 1H), 4.42-4.13 (m, 2H), 3.98-3.73 (m, 2H), 3.71-3.53 (m, 4H), 3.48-3.38 (m, 1H), 3.21-3.08 (m, 1H), 3.04-2.62 (m, 2H), 1.39-1.37 (m, 9H).
Step b. A solution of borane dimethyl sulfide complex (10 M in THF, 13.9 mL) was added dropwise to a solution of 1-(tert-butyl) 3-methyl 4-(((benzyloxy)carbonyl)-D-seryl)piperazine-1,3-dicarboxylate (6.5 g, 13.9 mmol) in THF (60 mL) at 0° C. The mixture was stirred at 25° C. for 12 h. Upon completion, MeOH (30 mL) was slowly added and the mixture was diluted with sat. aq. NaHCO3 (100 mL) and extracted with DCM (3×100 mL). The combined organic layers were washed with brine (300 mL), dried over Na2SO4 and evaporated. The residue was purified by reverse phase flash chromatography (water (0.1% NH4OH)/MeCN) to give 1-(tert-butyl) 3-methyl 4-((S)-2-(((benzyloxy)carbonyl)amino)-3-hydroxypropyl)piperazine-1,3-dicarboxylate (4.7 g, 74% yield) as a yellow solid.
m/z ES+ [M+H]+ 452.2; 1H NMR (400 MHz, DMSO-d6) δ ppm 7.41-7.27 (m, 5H), 6.95-6.85 (m, 1H), 5.07-4.94 (m, 2H), 4.66-4.57 (m, 1H), 4.06-3.82 (m, 1H), 3.77-3.49 (m, 5H), 3.46-3.35 (m, 2H), 3.13-3.09 (m, 1H), 3.05-2.75 (m, 2H), 2.72-2.61 (m, 1H), 2.60 (br s, 1H), 2.48-2.32 (m, 2H), 1.36 (s, 9H).
Step c. A mixture of 1-(tert-butyl) 3-methyl 4-((S)-2-(((benzyloxy)carbonyl)amino)-3-hydroxypropyl)piperazine-1,3-dicarboxylate (4.7 g, 10.4 mmol), 10% Pd/C (470 mg) in MeOH (80 mL) was stirred at 20° C. for 2 h under a H2 (15 psi) atmosphere. Upon completion, the reaction mixture was filtered and the filtrate was evaporated to give tert-butyl (7S)-7-(hydroxymethyl)-9-oxooctahydro-2H-pyrazino[1,2-a]pyrazine-2-carboxylate (2.97 g, crude) as an off-white solid.
1H NMR (400 MHz, DMSO-d6) δ ppm 7.91-7.58 (m, 1H), 4.85 (br s, 1H), 4.30-4.17 (m, 1H), 3.84 (br s, 1H), 3.47-3.40 (m, 1H), 3.32-3.26 (m, 1H), 3.19-3.08 (m, 2H), 3.01-2.93 (m, 1H), 2.87-2.72 (m, 2H), 2.43-2.35 (m, 2H), 2.15-2.01 (m, 1H), 1.40 (s, 9H).
Step d. A solution of borane dimethyl sulfide complex (10 M in THF, 6.7 mL) was added dropwise to a solution of tert-butyl (7S)-7-(hydroxymethyl)-9-oxooctahydro-2H-pyrazino[1,2-a]pyrazine-2-carboxylate (1.9 g, 6.66 mmol) in THF (30 mL) at 0° C. The mixture was stirred at 50° C. for 12 h. Upon completion, MeOH (20 mL) was slowly added to the reaction mixture, diluted with sat. aq. NaHCO3 (50 mL) and extracted with DCM (3×50 mL). The combined organic layers were washed with brine (120 mL), dried over Na2SO4 and evaporated to give tert-butyl (7S)-7-(hydroxymethyl)octahydro-2H-pyrazino[1,2-a]pyrazine-2-carboxylate (1.8 g, crude) as a yellow solid.
1H NMR (400 MHz, DMSO-d6) δ ppm 6.14-5.70 (m, 1H), 5.02-4.74 (m, 1H), 3.86-3.57 (m, 5H), 2.90-2.76 (m, 3H), 2.74-2.70 (m, 1H), 2.64-2.61 (m, 1H), 2.59-2.52 (m, 2H), 2.42-2.38 (m, 1H), 2.01-1.92 (m, 1H), 1.40 (s, 9H).
Step e. To a solution of tert-butyl (7S)-7-(hydroxymethyl)octahydro-2H-pyrazino[1,2-a]pyrazine-2-carboxylate (1.8 g, 6.63 mmol) and formaldehyde (37 wt. % in H2O; 2.69 g, 33.1 mmol) in MeOH (25 mL) was added NaBH3CN (833 mg, 13.2 mmol). The mixture was stirred at 20° C. for 12 h. Upon completion, the reaction mixture was evaporated and the residue was purified by reverse phase flash chromatography (water (0.1% NH4OH)/MeCN) to give tert-butyl (7S)-7-(hydroxymethyl)-8-methyloctahydro-2H-pyrazino[1,2-a]pyrazine-2-carboxylate (750 mg, 33% yield) as a white solid.
m/z ES+ [M+H]+ 286.2.
Step f. A solution of DMSO (657 mg, 8.41 mmol) in DCM (0.5 mL) was added dropwise to a solution of oxalyl chloride (533 mg, 4.20 mmol) in DCM (6 mL) at −78° C. The mixture was stirred at −78° C. for 10 min, after which a solution of tert-butyl (7S)-7-(hydroxymethyl)-8-methyloctahydro-2H-pyrazino[1,2-a]pyrazine-2-carboxylate (400 mg, 1.40 mmol) in DCM (2 mL) was added dropwise at −78° C. The mixture was stirred at −78° C. for 20 min, and then TEA (1.42 g, 14.0 mmol) was added at −78° C. The mixture was stirred and warmed to 20° C. for 30 min. Upon completion, the reaction mixture was diluted with sat. aq. NaHCO3 (10 mL) and the layers separated. The aqueous mixture as further extracted with DCM (2×5 mL). The combined organic layers were washed with brine (10 mL), dried over Na2SO4 and evaporated to give the title compound (360 mg, crude) as a brown solid.
m/z ES+ [M+H]+ 284.2.
Step a. A mixture of 3-methylpyrazin-2-amine (CAS: 19838-08-5; 3.00 g, 27.5 mmol) and ethyl 3-bromo-2-oxopropanoate (6.43 g, 33.0 mmol) in 1,2-dimethoxyethane (30 mL) was stirred at 25° C. for 16 h. Upon completion, the mixture was cooled to 0° C. and filtered. The filter cake was dissolved in EtOH (40 mL) and stirred at 80° C. for 2 h. Upon completion, the reaction mixture was cooled to 0° C. and filtered to give ethyl 8-methylimidazo[1,2-a]pyrazine-2-carboxylate (3.2 g, crude) as a white solid.
m/z ES+ [M+H]+ 205.9; 1H NMR (400 MHz, DMSO-d6) δ ppm 8.86 (s, 1H), 8.70 (d, J=4.8 Hz, 1H), 7.96 (d, J=4.8 Hz, 1H), 4.39-4.30 (m, 2H), 2.88 (s, 3H), 1.34 (t, J=7.2 Hz, 3H).
Step b. To a solution of ethyl 8-methylimidazo[1,2-a]pyrazine-2-carboxylate (1.2 g, 5.85 mmol) in EtOH (40 mL) was added 10% Pd/C (320 mg). The mixture was stirred at 50° C. for 16 h under a H2 (50 psi) atmosphere. Upon completion, the reaction mixture was filtered and evaporated to give ethyl 8-methyl-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazine-2-carboxylate (1.2 g, crude) as a yellow solid. To a mixture of ethyl 8-methyl-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazine-2-carboxylate (1.2 g, 5.73 mmol) and Na2CO3 (1.52 g, 14.3 mmol) in DCM (15 mL) was added Boc2O (1.88 g, 8.60 mmol). The reaction mixture was stirred at 20° C. for 2 h. Upon completion, the reaction mixture was quenched with water (30 mL) and extracted with EtOAc (3×20 mL). The combined organic layers were washed with brine (3×20 mL), dried over Na2SO4 and evaporated. The residue was purified by column chromatography (PE/EtOAc=1/1) to give 7-(tert-butyl) 2-ethyl 8-methyl-5,6-dihydroimidazo[1,2-a]pyrazine-2,7(8H)-dicarboxylate (1.1 g, 61% yield) as a white solid.
m/z ES+ [M+H]+ 310.4; 1H NMR (400 MHz, CDCl3) δ ppm 7.52 (s, 1H), 5.43-5.40 (m, 1H), 4.50-4.32 (m, 3H), 4.06-3.97 (m, 2H), 3.39-3.25 (m, 1H), 1.56 (d, J=6.8 Hz, 3H), 1.49 (s, 9H), 1.38 (t, J=7.2 Hz, 3H).
Step c. This step was conducted in a similar manner to Intermediate B3, step b.
1H NMR (400 MHz, CDCl3) δ ppm 9.86 (s, 1H), 7.53 (s, 1H), 5.43-5.39 (m, 1H), 4.52-4.39 (m, 1H), 4.14-3.94 (m, 2H), 3.39-3.26 (m, 1H), 1.58 (d, J=6.8 Hz, 3H), 1.51 (s, 9H).
The title compound was prepared in a similar manner to Intermediate B25, using 5-methylpyrazin-2-amine (CAS: 5521-58-4) in step a.
1H NMR (400 MHz, CDCl3) δ ppm 9.86 (s, 1H), 7.56 (s, 1H), 5.07 (d, J=17.6 Hz, 1H), 4.94-4.82 (m, 1H), 4.35 (d, J=17.6 Hz, 1H), 4.21 (dd, J=12.4, 4.8 Hz, 1H), 3.90 (d, J=12.4 Hz, 1H), 1.50 (s, 9H), 1.21 (d, J=7.2 Hz, 3H).
Step a. A solution of ethyl 2-diazoacetate (8.50 g, 74.5 mmol) in DCM (80 mL) at 10° C. was added dropwise over 2 h to a solution of tert-butyl 4-hydroxypiperidine-1-carboxylate (5.0 g, 24.8 mmol) and Rh(OAc)2 (275 mg, 1.24 mmol) in DCM (80 mL). The mixture was stirred at 20° C. for 46 h. Upon completion, the reaction mixture was evaporated and the residue was purified by column chromatography (PE/EtOAc=8/1) to give tert-butyl 4-(2-ethoxy-2-oxoethoxy)piperidine-1-carboxylate (3 g, 40% yield) as a light yellow oil.
1H NMR (400 MHz, CDCl3) δ ppm 4.25-4.20 (m, 2H), 4.12 (s, 2H), 3.84-3.76 (m, 2H), 3.61-3.53 (m, 1H), 3.12-3.07 (m, 2H), 1.91-1.82 (m, 2H), 1.60-1.57 (m, 2H), 1.46 (s, 9H), 1.30 (t, J=7.2 Hz, 3H).
Step b. This step was conducted in a similar manner to Intermediate B3, step b.
1H NMR (400 MHz, CDCl3) δ ppm 9.74 (s, 1H), 3.85-3.71 (m, 2H), 3.58-3.51 (m, 2H), 3.14-3.01 (m, 2H), 1.95-1.80 (m, 2H), 1.69-1.65 (m, 1H), 1.63-1.53 (m, 2H), 1.46 (s, 9H).
The title compound was prepared in a similar manner to Intermediate B27, using tert-butyl (S)-3-(hydroxymethyl)pyrrolidine-1-carboxylate (CAS: 199174-24-8) in step a.
1H NMR (400 MHz, CDCl3) δ ppm 9.73 (s, 1H), 4.10 (s, 2H), 3.59-3.51 (m, 3H), 3.47-3.43 (m, 1H), 3.35-3.31 (m, 1H), 3.15-3.10 (m, 1H), 2.55-2.50 (m, 1H), 2.04-1.94 (m, 1H), 1.78-1.67 (m, 1H), 1.47 (s, 9H).
The title compound was prepared in a similar manner to Intermediate B27, using tert-butyl (R)-3-(hydroxymethyl)pyrrolidine-1-carboxylate (CAS: 138108-72-2) in step a.
1H NMR (400 MHz, CDCl3) δ ppm 9.72 (s, 1H), 4.09 (s, 2H), 3.47-3.43 (m, 4H), 3.34-3.30 (m, 1H), 3.13-3.08 (m, 1H), 2.54-2.49 (m, 1H), 2.02-1.96 (m, 2H), 1.46 (s, 9H).
Step a. To a solution of oxalyl chloride (946 mg, 7.45 mmol) in DCM (5 mL) was added DMSO (1.16 g, 14.9 mmol) at −78° C. The mixture was stirred at −78° C. for 10 min. A solution of tert-butyl 3-(2-hydroxyethyl)azetidine-1-carboxylate (CAS: 152537-03-6; 500 mg, 2.48 mmol) in DCM (4 mL) was then added and the mixture was stirred at −78° C. for a further 30 min. After which, TEA (2.51 g, 24.8 mmol) was added and the mixture was stirred at −78° C. for an additional 30 min. Upon completion, the reaction mixture was warmed to 20° C., diluted with water (50 mL) and extracted with DCM (20 mL). The combined organic layers were washed with brine (50 mL), dried over Na2SO4 and evaporated to give tert-butyl 3-(2-oxoethyl)azetidine-1-carboxylate (500 mg, crude) as a colourless liquid.
1H NMR (400 MHz, CDCl3) δ ppm 9.78 (s, 1H), 4.13 (t, J=8.4 Hz, 2H), 3.57 (dd, J=8.8, 5.6 Hz, 2H), 2.99-2.87 (m, 1H), 2.86-2.80 (m, 2H), 1.44 (s, 9H).
Step b. To a solution of tert-butyl 3-(2-oxoethyl)azetidine-1-carboxylate (500 mg, 2.51 mmol) in DCM (10 mL) was added ethyl 2-(triphenyl-λ5-phosphanylidene)acetate (CAS: 1099-45-2; 962 mg, 2.76 mmol). The mixture was stirred at 20° C. for 16 h. Upon completion, the reaction mixture was filtered and evaporated. The residue was purified by column chromatography (E/EtOAc=5/1) to give tert-butyl (E)-3-(4-ethoxy-4-oxobut-2-en-1-yl)azetidine-1-carboxylate (440 mg, 63% yield) as a colourless liquid.
1H NMR (400 MHz, CDCl3) δ ppm 6.90-6.82 (m, 1H), 5.80 (d, J=15.6 Hz, 1H), 4.23-4.16 (m, 2H), 4.06-4.02 (t, J=8.4 Hz, 2H), 3.60-3.55 (m, 2H), 2.71-2.60 (m, 1H), 2.50 (t, J=6.8 Hz, 2H), 1.44 (s, 9H), 1.30 (t, J=7.2 Hz, 3H).
Step c. To a solution of tert-butyl (E)-3-(4-ethoxy-4-oxobut-2-en-1-yl)azetidine-1-carboxylate (440 mg, 1.63 mmol) in EtOH (5 mL) was added 10% Pd/C (50 mg). The mixture was stirred at 25° C. for 16 h under a H2 atmosphere (50 psi). Upon completion, the reaction mixture was filtered and evaporated to give tert-butyl 3-(4-ethoxy-4-oxobutyl)azetidine-1-carboxylate (400 mg, crude) as a colourless liquid.
1H NMR (400 MHz, CDCl3) δ ppm 4.17-4.11 (m, 2H), 4.02-3.97 (m, 2H), 3.56-3.52 (m, 2H), 2.30 (t, J=6.8 Hz, 2H), 1.67-1.58 (m, 5H), 1.44 (s, 9H), 1.27 (t, J=7.2 Hz, 3H).
Step d. This step was conducted in a similar manner to Intermediate B3, step b.
1H NMR (400 MHz, CDCl3) δ ppm 9.78 (t, J=1.6 Hz, 1H), 4.01 (t, J=8.4 Hz, 2H), 3.54 (dd, J=8.8, 5.6 Hz, 2H), 2.47 (m, 3H), 1.65-1.55 (m, 4H), 1.44 (s, 9H).
Step a. This step was conducted in a similar manner to Intermediate B27, step a, using tert-butyl 3-(hydroxymethyl)azetidine-1-carboxylate (CAS: 142253-56-3)
Step b. To a solution of tert-butyl 3-((2-ethoxy-2-oxoethoxy)methyl)azetidine-1-carboxylate (300 mg, 1.10 mmol) in THF (3 mL) was added LiBH4 (72 mg, 3.29 mmol) at 0° C. The mixture was stirred at 0-20° C. for 16 h. Upon completion, the reaction mixture was quenched with sat. aq. NH4Cl (1 mL) at 0° C., filtered and evaporated to give tert-butyl 3-((2-hydroxyethoxy)methyl)azetidine-1-carboxylate (210 mg, crude) as a colourless oil.
m/z ES+ [M+H]+ 232.2; 1H NMR (400 MHz, CDCl3) δ ppm 4.01 (t, J=8.4 Hz, 2H), 3.77-3.72 (m, 2H), 3.68-3.64 (m, 2H), 3.63 (d, J=6.8 Hz, 2H), 3.60-3.56 (m, 2H), 2.83-2.70 (m, 1H), 1.95 (br s, 1H), 1.45 (s, 9H).
Step c. To a solution of oxalyl chloride (346 mg, 2.72 mmol) in DCM (10 mL) was added DMSO (426 mg, 5.45 mmol) in DCM (5 mL) at −78° C. The mixture was stirred at −70° C. for 15 min. A mixture of tert-butyl 3-((2-hydroxyethoxy)methyl)azetidine-1-carboxylate (210 mg, 0.91 mmol) in DCM (5 mL) was then added and the mixture was stirred at −70° C. for a further 2 h 30 min. After which, TEA (827 mg, 8.17 mmol) was added and the reaction mixture was stirred at −70° C. for a further 30 min. Upon completion, the reaction mixture was diluted with water (5 mL) and extracted with DCM (3×5 mL). The combined organic layers were dried over Na2SO4 and evaporated to give the title compound (180 mg, crude) as a light yellow oil.
1H NMR (400 MHz, CDCl3) δ ppm 9.72 (s, 1H), 4.12 (s, 2H), 4.06-3.99 (m, 2H), 3.73-3.68 (m, 2H), 3.14-3.11 (m, 2H), 2.87-2.76 (m, 1H), 1.44 (s, 9H).
The title compound was prepared in a similar manner to to Intermediate B3, step b, using methyl 3-((tert-butoxycarbonyl)amino)bicyclo[1.1.1]pentane-1-carboxylate (CAS: 676371-64-5).
1H NMR (400 MHz, CDCl3) δ ppm 9.67 (s, 1H), 5.00 (brs, 1H), 2.29 (s, 6H), 1.46 (s, 9H).
Step a. A mixture of dimethyl 1H-pyrazole-3,5-dicarboxylate (5.0 g, 27.2 mmol), tert-butyl (2-bromoethyl)carbamate (7.30 g, 32.6 mmol) and K2CO3 (5.6 g, 40.7 mmol) in DMF (100 mL) was stirred at 0° C. for 2 h under a N2 atmosphere. Upon completion, the reaction mixture was quenched with water (100 mL) and filtered. The filter cake was washed with water (50 mL) to give dimethyl 1-(2-((tert-butoxycarbonyl)amino)ethyl)-1H-pyrazole-3,5-dicarboxylate (8.50 g, 96% yield) as a yellow solid.
m/z ES+ [M+H]+ 328.1; 1H NMR (400 MHz, CDCl3) δ ppm 7.38 (s, 1H), 4.79-4.75 (m, 3H), 3.95 (s, 3H), 3.93 (s, 3H), 3.64 (d, J=4.0 Hz, 2H), 1.39 (s, 9H).
Step b. A mixture of dimethyl 1-(2-((tert-butoxycarbonyl)amino)ethyl)-1H-pyrazole-3,5-dicarboxylate (1 g, 3.06 mmol) and TFA (6.97 g, 61.1 mmol) in DCM (10 mL) was stirred at 25° C. for 1 h under a N2 atmosphere. Upon completion, the mixture was evaporated to give dimethyl 1-(2-aminoethyl)-1H-pyrazole-3,5-dicarboxylate as a TFA salt (1.0 g, 96% yield) as a yellow oil.
m/z ES+ [M+H]+ 228.0.
Step c. A mixture of dimethyl 1-(2-aminoethyl)-1H-pyrazole-3,5-dicarboxylate (TFA salt, 1.0 g, 2.93 mmol) and TEA (2.97 g, 29.3 mmol, 4.08 mL) in MeCN (100 mL) was stirred at 60° C. for 12 h under a N2 atmosphere. Upon completion, the reaction mixture was diluted with water (100 mL) and filtered to give methyl 4-oxo-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazine-2-carboxylate (500 mg, 87% yield) as a white solid.
m/z ES+ [M+H]+ 196.1; 1H NMR (400 MHz, DMSO-d6) δ ppm 8.43 (s, 1H), 7.10 (s, 1H), 4.40 (s, 2H), 3.82 (s, 3H), 3.64 (s, 2H).
Step d. A mixture of methyl 4-oxo-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazine-2-carboxylate (500 mg, 2.56 mmol) and a solution of borane dimethyl sulfide complex (10 M in THF, 1.28 mL) in THF (5 mL) was stirred at 25° C. for 12 h under a N2 atmosphere. Upon completion, the reaction mixture was quenched with MeOH (10 mL) at 0° C. and stirred for a further 2 h. The mixture was evaporated to give methyl 4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazine-2-carboxylate (0.5 g, crude) as a yellow oil.
m/z ES+ [M+H]+ 182.1.
Step e. To a solution of methyl 4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazine-2-carboxylate (8.0 g, 44.1 mmol) in DCM (80 mL) was added Boc2O (14.4 g, 66.2 mmol) and TEA (13.4 g, 132 mmol). The reaction mixture was stirred at 25° C. for 1 h. Upon completion, the mixture was diluted with water (100 mL) and extracted with EtOAc (3×40 mL). The combined organic layers were dried over Na2SO4 and evaporated. The residue was purified by column chromatography (PE/EtOAc=1/1) to give 5-(tert-butyl) 2-methyl 6,7-dihydropyrazolo[1,5-a]pyrazine-2,5(4H)-dicarboxylate (2.5 g, 19% yield) as a white solid.
m/z ES+ [M+H]+ 282.1; 1H NMR (400 MHz, CDCl3) δ ppm 6.63 (s, 1H), 4.68 (s, 2H), 4.26 (t, J=5.6 Hz, 2H), 3.93 (s, 4H), 3.92-3.89 (m, 1H), 1.50 (s, 9H).
Steps f-g. These 2 steps were conducted in a similar manner to Intermediate B5, steps b-c.
m/z ES+ [M+H]+ 251.8.
Step a. Oxalyl chloride (4.48 g, 35.3 mmol) and DMF (380 mg, 5.20 mmol) were added to a solution of 3-(methoxycarbonyl)bicyclo[1.1.1]pentane-1-carboxylic acid (CAS: 83249-10-9; 5.00 g, 29.4 mmol) in DCM (50 mL). The mixture was stirred at 25° C. for 2 h. Upon completion, the reaction mixture was evaporated to give the acid chloride (5.5 g, crude) as a white solid. Ammonia gas was bubbled through DCM (40 mL) for 30 mins, after which, a solution of the acid chloride in DCM (10 mL) was added. The reaction mixture was stirred at 25° C. for 2 h. Upon completion, the reaction mixture was diluted with water (50 mL) and extracted with EtOAc (3×20 mL). The combined organic layers were dried over Na2SO4, filtered and evaporated. The residue was purified by column chromatography (PE/EtOAc=1/1) to give methyl 3-carbamoylbicyclo[1.1.1]pentane-1-carboxylate (3.7 g, 75% yield) as a white solid.
1H NMR (400 MHz, DMSO-d6) δ ppm 7.35 (s, 1H), 7.04 (s, 1H), 3.60 (s, 3H), 2.12 (s, 6H).
Step b. To a mixture of methyl 3-carbamoylbicyclo[1.1.1]pentane-1-carboxylate (1.5 g, 8.9 mmol) in THF (20 mL) was added a solution of borane dimethyl sulfide complex (10 M in THF, 2.66 mL). The reaction mixture was stirred at 25° C. for 4 h under a N2 atmosphere. Upon completion, the reaction mixture was quenched with MeOH (5 mL) at 0° C., filtered and evaporated to give methyl 3-(aminomethyl)bicyclo[1.1.1]pentane-1-carboxylate (1.4 g, crude) as a white solid which was used without further purification.
Step c. To a solution of methyl 3-(aminomethyl)bicyclo[1.1.1]pentane-1-carboxylate (1.4 g, 9.02 mmol) in THF (10 mL) and water (10 mL) was added TEA (2.74 g, 27.1 mmol) and Boc2O (2.95 g, 13.5 mmol). The mixture was stirred at 25° C. for 16 h. Upon completion, the reaction mixture was diluted with water (30 mL) and extracted with EtOAc (3×50 mL). The combined organic layers were dried over Na2SO4, and evaporated. The residue was purified by column chromatography (PE/EtOAc=3/1) to give methyl 3-(((tert-butoxycarbonyl)amino)methyl)bicyclo[1.1.1]pentane-1-carboxylate (400 mg, 17% yield) as a light yellow oil.
1H NMR (400 MHz, CDCl3) δ ppm 4.54 (br s, 1H), 3.66 (s, 3H), 3.26-3.17 (m, 2H), 1.95 (s, 6H), 1.43 (s, 9H).
Step d. This step was conducted in a similar manner to Intermediate B1, step b.
1H NMR (400 MHz, CDCl3) δ ppm 4.68 (s, 1H), 4.51 (brs, 1H), 3.59 (s, 2H), 3.20 (d, J=5.6 Hz, 2H), 1.61 (s, 6H), 1.44 (s, 9H).
Step e. To a solution of oxalyl chloride (212 mg, 1.67 mmol) in DCM (3 mL) was added DMSO (261 mg, 3.34 mmol) in DCM (3 mL) at −78° C. The mixture was stirred at −60° C. for 15 min, A mixture of methyl 3-(((tert-butoxycarbonyl)amino)methyl)bicyclo[1.1.1]pentane-1-carboxylate (190 mg, 0.84 mmol) in DCM (5 mL) was then added. The mixture was stirred at −60° C. for a further 2 h 30 min. After which, TEA (507 mg, 5.02 mmol) was added and the reaction mixture was stirred at −60° C. for a further 30 min. Upon completion, the reaction mixture was diluted with water (15 mL) and extracted with DCM (3×10 mL). The combined organic layers were dried over Na2SO4 and evaporated to give the title compound (180 mg, crude) as a colourless oil which was used without further purification.
1H NMR (400 MHz, CDCl3) δ ppm 9.57 (s, 1H), 4.54 (br s, 1H), 3.23 (d, J=5.6 Hz, 2H), 1.95 (s, 6H), 1.45 (s, 9H).
Step a. To a solution of 2-(2-aminoethoxy)ethanol (CAS: 929-06-6; 10.0 g, 95.1 mmol) in toluene (100 mL) was added isobenzofuran-1,3-dione (14.1 g, 95.1 mmol). The reaction mixture was stirred at 110° C. for 4 h. Upon completion, the reaction mixture was evaporated. The residue was triturated with PE/EtOAc (5/1; 40 mL) and filtered to give 2-(2-(2-hydroxyethoxy)ethyl)isoindoline-1,3-dione (21 g, 92% yield) as a white solid.
1H NMR (400 MHz, CDCl3) δ ppm 7.88-7.82 (m, 2H), 7.76-7.69 (m, 2H), 3.95-3.87 (m, 2H), 3.79-3.72 (m, 2H), 3.71-3.66 (m, 2H), 3.63-3.56 (m, 2H), 2.80 (br s, 1H).
Step b. To a solution of oxalyl chloride (3.24 g, 25.5 mmol) in DCM (30 mL) was added a solution of DMSO (3.99 g, 51.0 mmol) in DCM (5 mL) dropwise at −78° C. After stirring for 10 min, a solution of 2-(2-(2-hydroxyethoxy)ethyl)isoindoline-1,3-dione (2 g, 8.50 mmol) in DCM (20 mL) was added dropwise. The mixture was stirred at −78° C. for 30 min, after which TEA (7.74 g, 76.5 mmol) was added. The reaction mixture was stirred at −78° C. for a further 30 min. Upon completion, the reaction mixture was warmed to 20° C. and water (50 mL) was added. The layers were separated and the organic layer was washed with brine (50 mL), dried over Na2SO4 and evaporated to give the title compound (1.5 g, 76% yield) as a yellow oil.
1H NMR (400 MHz, CDCl3) δ ppm 9.66 (s, 1H), 7.93-7.78 (m, 2H), 7.77-7.62 (m, 2H), 4.14-4.12 (m, 1H), 4.00-3.94 (m, 1H), 3.94-3.86 (m, 1H), 3.85-3.81 (m, 1H), 3.80-3.65 (m, 1H), 3.63-3.39 (m, 1H).
The title compound was prepared in a similar manner to Intermediate B35, using 5-aminopentan-1-ol (CAS: 2508-29-4) in step a.
m/z ES+ [M+H]+ 232.1 1H NMR (400 MHz, CDCl3) δ ppm 9.77 (t, J=1.6 Hz, 1H), 7.89-7.82 (m, 2H), 7.76-7.70 (m, 2H), 3.72 (t, J=6.8 Hz, 2H), 2.54-2.50 (m, 2H), 1.79-1.65 (m, 4H).
Step a. To a solution of 3-hydroxy-4-nitrobenzaldehyde (CAS: 704-13-2; 1.00 g, 6.0 mmol) in DMF (12 mL) was added K2CO3 (910 mg, 6.6 mmol) and MeI (1.70 g, 12.0 mmol). The mixture was stirred at 20° C. for 2 h. Upon completion, the reaction mixture was diluted with water (50 mL) and extracted with EtOAc (3×50 mL). The combined organic layers were washed with water (2×100 mL), brine (2×200 mL), dried over Na2SO4 and evaporated. The residue was purified by column chromatography (PE/EtOAc=6/1) to give the title compound (1.1 g, 99% yield) as a yellow solid.
1H NMR (400 MHz, CDCl3) δ ppm 10.07 (s, 1H), 7.94 (d, J=8.0 Hz, 1H), 7.61 (d, J=1.2 Hz, 1H), 7.55 (dd, J=8.0, 1.6 Hz, 1H), 4.05 (s, 3H).
Step a. To a solution of oxalyl chloride (1.63 g, 12.9 mmol) in DCM (5 mL) was added DMSO (2.01 g, 25.7 mmol) in DCM (5 mL) at −78° C. The mixture was stirred at −70° C. for 15 min, after which, a mixture of tert-butyl 4-fluoro-4-(hydroxymethyl)piperidine-1-carboxylate (CAS: 614730-97-1; 1.5 g, 6.43 mmol) in DCM (5 mL) was added and the mixture was stirred at −70° C. for a further 2 h 30 min. After this, TEA (3.90 g, 38.6 mmol) was added and the reaction mixture was stirred at −70° C. for a further 30 min. Upon completion, the reaction mixture was diluted with water (40 mL) and extracted with DCM (2×20 mL). The combined organic layers were washed with brine (3×20 mL), dried over with Na2SO4 and evaporated. The residue was purified by column chromatography (PE/EtOAc=3/1) to give the title compound (1.1 g, 74% yield) as a light yellow oil.
1H NMR (400 MHz, CDCl3) δ ppm 9.73 (d, J=5.2 Hz, 1H), 4.05-3.95 (m, 2H), 3.17-2.97 (m, 2H), 1.94-1.79 (m, 2H), 1.73-1.55 (m, 2H), 1.46 (s, 9H).
Step a. To a solution of 1-(tert-butyl) 4-ethyl 3-oxopiperidine-1,4-dicarboxylate (CAS: 71233-25-5; 10 g, 36.9 mmol) in EtOH (60 mL) was added NaBH4 (697 mg, 18.4 mmol). The reaction mixture was stirred at 20° C. for 2 h. Upon completion, the reaction mixture was slowly poured into sat. aq. NH4Cl (50 mL) and extracted with EtOAc (3×50 mL). The combined organic layers were washed with brine (50 mL), dried over Na2SO4 and evaporated. The crude product was purified by column chromatography (PE/EtOAc=5/1) to give 1-(tert-butyl) 4-ethyl 3-hydroxypiperidine-1,4-dicarboxylate (7.5 g, 74% yield) as a colourless oil.
1H NMR (400 MHz, CDCl3) δ ppm 4.31-4.15 (m, 3H), 4.07-3.97 (m, 2H), 3.04-2.91 (m, 1H), 2.89-2.79 (m, 1H), 2.78-2.59 (m, 1H), 2.58-2.49 (m, 1H), 2.12-1.99 (m, 1H), 1.79-1.68 (m, 1H), 1.48-1.45 (m, 9H), 1.30-1.25 (m, 3H).
Step b. To a solution of 1-(tert-butyl) 4-ethyl 3-hydroxypiperidine-1,4-dicarboxylate (1.5 g, 5.49 mmol) and imidazole (1.12 g, 16.46 mmol) in DCM (15 mL) was added TBDPSCl (1.96 g, 7.13 mmol) at 0° C. The reaction mixture was stirred at 10° C. for 12 h. Upon completion, the mixture was filtered and evaporated. The residue was purified by reverse phase flash chromatography (water (0.1% FA)/MeCN) to give 1-(tert-butyl) 4-ethyl 3-((tert-butyldiphenylsilyl)oxy)piperidine-1,4-dicarboxylate (1.9 g, 64% yield) as a yellow oil.
m/z ES+ [M+H]+ 512.2.
Step c. This step was conducted in a similar manner to Intermediate B3, step b.
1H NMR (400 MHz, CDCl3) δ ppm 9.44-9.40 (m, 1H), 7.69-7.53 (m, 5H), 7.44-7.25 (m, 5H), 4.27-4.12 (m, 1H), 3.87-3.53 (m, 2H), 3.26-2.99 (m, 1H), 2.95-2.75 (m, 1H), 2.45-2.20 (m, 1H), 1.80-1.45 (m, 2H), 1.30-1.23 (m, 9H), 1.06-0.94 (m, 9H).
Step a. To a solution of methyl 6-cyanonicotinate (CAS: 89809-65-4; 1 g, 6.17 mmol) and 10% Pd/C (0.2 g) in MeOH (20 mL) was added HCl (729 mg, 7.40 mmol). The reaction mixture was stirred at 20° C. for 15 h under a H2 atmosphere (15 psi). Upon completion, the mixture was filtered and the filtrate was evaporated to give methyl 6-(aminomethyl)nicotinate as the HCl salt (1.2 g, 96% yield) as a yellow oil.
1H NMR (400 MHz, DMSO-d6) δ ppm 9.07 (d, J=1.6 Hz, 1H), 8.73 (br s, 3H), 8.38-8.30 (m, 1H), 7.70 (d, J=8.4 Hz, 1H), 4.30-4.20 (m, 2H), 3.89 (s, 3H).
Step b. To a solution of methyl 6-(aminomethyl)nicotinate hydrochloride (1.2 g, 5.92 mmol) and TEA (1.80 g, 17.8 mmol) in MeOH (20 mL) was added Boc2O (1.94 g, 8.88 mmol). The reaction mixture was stirred at 20° C. for 12 h. Upon completion, the mixture was evaporated, water (50 mL) was added and the aqueous mixture was extracted with EtOAc (2×50 mL). The combined organic layers were evaporated. The residue was purified by column chromatography (PE/EtOAc=6/1) to give methyl 6-(((tert-butoxycarbonyl)amino)methyl)nicotinate (1.4 g, 89% yield) as a white solid.
1H NMR (400 MHz, CDCl3) δ ppm 9.15 (d, J=2.0 Hz, 1H), 8.41-8.29 (m, 1H), 7.46 (d, J=8.0 Hz, 1H), 5.66 (br s, 1H), 4.56 (d, J=5.2 Hz, 2H), 3.97 (s, 3H), 1.46 (s, 9H).
Step c. This step was conducted in a similar manner to Intermediate B3, step b.
1H NMR (400 MHz, CDCl3) δ ppm 10.11 (s, 1H), 9.00 (d, J=1.6 Hz, 1H), 8.16 (dd, J=8.0, 2.0 Hz, 1H), 7.47 (d, J=8.0 Hz, 1H), 5.55 (br s, 1H), 4.55 (d, J=5.2 Hz, 2H), 1.48 (s, 9H).
Step a. To a solution of (R)-2-(1-(tert-butoxycarbonyl)pyrrolidin-3-yl)acetic acid (CAS: 204688-60-8; 1.00 g, 4.36 mmol) in THF (10 mL) was added a solution of borane tetrahydrofuran complex (1 M in THF, 5.23 mL) at 0° C. The reaction mixture was stirred at 20° C. for 1 h. Upon completion, the reaction mixture was quenched with 10% aq. NaOH (2 mL). The reaction mixture was diluted with water (30 mL) and extracted with EtOAc (30 mL). The organic layer was dried over Na2SO4 and evaporated to give tert-butyl (R)-3-(2-hydroxyethyl)pyrrolidine-1-carboxylate (0.93 g, 99% yield) as a colourless oil.
1H NMR (400 MHz, CDCl3) δ ppm 3.74-3.66 (m, 2H), 3.60-3.52 (m, 1H), 3.50-3.41 (m, 1H), 3.31-3.21 (m, 1H), 2.94-2.86 (m, 1H), 2.34-2.19 (m, 1H), 2.07-1.97 (m, 1H), 1.71-1.63 (m, 2H), 1.58-1.49 (m, 1H), 1.46 (s, 9H), 1.45-1.42 (m, 1H).
Step b. To a solution of tert-butyl (R)-3-(2-hydroxyethyl)pyrrolidine-1-carboxylate (0.3 g, 1.39 mmol) in DCM (3 mL) was added Dess-Martin periodinane (1.18 g, 2.79 mmol). The reaction mixture was stirred at 20° C. for 1 h. Upon completion, the pH of the reaction mixture was adjusted to ˜pH 9 with sat. aq. NaHCO3 (20 mL). The layers were separated and the organic layer was washed with brine (20 mL), dried over Na2SO4, evaporated and purified by column chromatography (PE/EtOAc=3/1) to give the title compound (150 mg, 50% yield) as a colourless oil.
1H NMR (400 MHz, CDCl3) δ ppm 9.79 (s, 1H), 3.62-3.56 (m, 1H), 3.50-3.42 (m, 1H), 3.34-3.30 (m, 1H), 3.00-2.85 (m, 1H), 2.68-2.57 (m, 3H), 2.11-2.08 (m, 1H), 1.56-1.49 (m, 1H), 1.48 (s, 9H).
The title compound was prepared in a similar manner to Intermediate B41, using tert-butyl (S)-3-(hydroxymethyl)pyrrolidine-1-carboxylate (CAS: 204688-61-9) in step a.
1H NMR (400 MHz, CDCl3) δ ppm 9.80 (s, 1H), 3.65-3.29 (m, 3H), 3.02-2.85 (m, 1H), 2.66-2.54 (m, 3H), 2.12-2.07 (m, 1H), 1.56-1.50 (m, 1H), 1.46 (s, 9H).
Step a. This step was conducted in a similar manner to Intermediate B41, step b, using tert-butyl (S)-3-(hydroxymethyl)pyrrolidine-1-carboxylate (CAS: 199174-24-8).
1H NMR (400 MHz, CDCl3) δ ppm 9.70 (s, 1H), 3.80-3.63 (m, 1H), 3.59-3.46 (m, 1H), 3.43-3.40 (m, 2H), 3.10-2.95 (m, 1H), 2.29-2.07 (m, 2H), 1.47 (s, 9H).
The title compound was prepared in a similar manner to Intermediate B43, using tert-butyl (R)-3-(hydroxymethyl)pyrrolidine-1-carboxylate (CAS: 138108-72-2).
1H NMR (400 MHz, CDCl3) δ ppm 9.69 (s, 1H), 3.79-3.63 (m, 1H), 3.55-3.45 (m, 1H), 3.43-3.30 (m, 2H), 3.10-2.95 (m, 1H), 2.20-2.04 (m, 2H), 1.47 (s, 9H).
The title compound was prepared in a similar manner to Intermediate B43, using tert-butyl 4-(3-hydroxypropyl)piperidine-1-carboxylate (CAS: 156185-63-6).
1H NMR (400 MHz, CDCl3) δ ppm 9.79 (t, J=1.6 Hz, 1H), 4.17-4.00 (m, 2H), 2.70-2.62 (m, 2H), 2.52-2.43 (m, 2H), 1.68-1.56 (m, 4H), 1.45 (s, 9H), 1.44-1.34 (m, 1H), 1.13-1.08 (m, 2H).
Step a. To a solution of benzyl piperazine-1-carboxylate (3.00 g, 13.6 mmol) and 1-hydroxypropan-2-one (1.11 g, 15.0 mmol) in MeOH (40 mL) was added acetic acid (82 mg, 1.36 mmol) and 4 Å molecular sieves (1 g). The mixture was stirred at 15° C. for 30 min, after which, NaBH3CN (2.57 g, 40.9 mmol) was added. The reaction mixture was stirred at 15° C. for 90 min. Upon completion, the reaction mixture was filtered and the filtrate was evaporated. Sat. aq. NaHCO3 (10 mL) was added and the aqueous mixture was extracted with EtOAc (3×20 mL). The combined organic layers were washed with brine (10 mL), dried over Na2SO4 and evaporated. The crude product was purified by column chromatography (PE/EtOAc=5/1) to give benzyl 4-(1-hydroxypropan-2-yl)piperazine-1-carboxylate (1.7 g, 43% yield) as a yellow oil.
m/z ES+ [M+H]+ 279.2; 1H NMR (400 MHz, CDCl3) δ ppm 7.40-7.31 (m, 5H), 5.12 (s, 2H), 3.64-3.45 (m, 6H), 2.81-2.75 (m, 1H), 2.69-2.59 (m, 3H), 2.39-2.24 (m, 2H), 0.90 (d, J=6.8 Hz, 3H).
Step b. DMSO (505 mg, 6.47 mmol) was added to a solution of oxalyl chloride (410 mg, 3.23 mmol) in DCM (5 mL) at −78° C. The mixture was stirred at −78° C. for 10 min. After which, a solution of benzyl 4-(1-hydroxypropan-2-yl)piperazine-1-carboxylate (300 mg, 1.08 mmol) in DCM (2 mL) was added. The mixture was stirred at −78° C. for a further 30 min, and then TEA (1.09 g, 10.8 mmol) was added. The mixture was stirred at −78° C. for a further 60 min. Upon completion, the reaction mixture was quenched with water (30 mL) and extracted with EtOAc (3×25 mL). The combined organic layers were washed with brine (25 mL), dried over Na2SO4 and evaporated to give the title compound (300 mg, crude) as a yellow oil.
1H NMR (400 MHz, CDCl3) δ ppm 9.67 (s, 1H), 7.39-7.30 (m, 5H), 5.14 (s, 2H), 3.62-3.50 (m, 4H), 3.13-3.09 (m, 1H), 2.61-2.45 (m, 4H), 1.15 (d, J=6.8 Hz, 3H).
Step a. To a solution of 1-(tert-butyl) 4-ethyl 3-oxopiperidine-1,4-dicarboxylate (CAS: 71233-25-5; 10 g, 36.9 mmol) in EtOH (60 mL) was added NaBH4 (697 mg, 18.4 mmol). The reaction mixture was stirred at 20° C. for 2 h. Upon completion, the reaction mixture was slowly added into sat. aq. NH4Cl (50 mL) and the aqueous mixture was extracted with EtOAc (3×50 mL). The combined organic layers were washed with brine (50 ml), dried over Na2SO4, evaporated and purified by column chromatography (PE/EtOAc=5/1) to give 1-(tert-butyl) 4-ethyl 3-hydroxypiperidine-1,4-dicarboxylate (7.5 g, 74% yield) as a colourless oil.
1H NMR (400 MHz, CDCl3) δ ppm 4.31-4.15 (m, 4H), 4.07-3.97 (m, 1H), 3.04-2.91 (m, 1H), 2.89-2.79 (m, 1H), 2.78-2.59 (m, 1H), 2.58-2.49 (m, 1H), 2.12-1.99 (m, 1H), 1.79-1.68 (m, 1H), 1.48-1.45 (m, 9H), 1.30-1.25 (m, 3H).
Step b. To a solution of 1-(tert-butyl) 4-ethyl 3-hydroxypiperidine-1,4-dicarboxylate (7.5 g, 27.4 mmol) and TEA (11.1 g, 110 mmol) in DCM (75 mL) was added MsCl (6.29 g, 54.9 mmol) at 0° C. The reaction mixture was stirred at 20° C. for 12 h. Upon completion, the reaction mixture was diluted with water (50 mL) and extracted with DCM (100 mL). The organic layer was washed with brine (50 mL), dried over Na2SO4 and evaporated to give 1-(tert-butyl) 4-ethyl 3-((methylsulfonyl)oxy)piperidine-1,4-dicarboxylate (9.6 g, crude) as a yellow oil.
m/z ES+ [M+Na]+ 374.0.
Step c. A mixture of 1-(tert-butyl) 4-ethyl 3-((methylsulfonyl)oxy)piperidine-1,4-dicarboxylate (9.6 g, 27.3 mmol), TBAF (1 M in THF, 41.0 mL) and trimethylsilyl cyanide (4.07 g, 41.0 mmol) in MeCN (90 mL) was stirred at 80° C. for 12 h. Upon completion, the reaction mixture was diluted with water (40 mL) and extracted with EtOAc (3×40 mL). The combined organic layers were washed with brine (40 mL), dried over Na2SO4 and evaporated. The residue was purified by column chromatography (PE/EtOAc=5/1) to give two isomers.
The first eluting isomer was confirmed by 2D NMR to be Intermediate B47b: 1-(tert-butyl) 4-ethyl (rac-trans)-3-cyanopiperidine-1,4-dicarboxylate (1.4 g, 18% yield) as a yellow oil.
1H NMR (400 MHz, CDCl3) δ ppm 4.27-4.21 (m, 3H), 3.91-3.87 (m, 1H), 3.28-3.10 (m, 1H), 3.09-3.03 (m, 1H), 3.00-2.92 (m, 1H), 2.78-2.74 (m, 1H), 2.06-2.04 (m, 1H), 1.70-1.65 (m, 1H), 1.47 (s, 9H), 1.32-1.27 (m, 3H).
The second eluting isomer was confirmed by 2D NMR to be Intermediate B47a: 1-(tert-butyl) 4-ethyl (rac-cis)-3-cyanopiperidine-1,4-dicarboxylate (5 g, crude) as a yellow oil.
1H NMR (400 MHz, CDCl3) δ ppm 4.51-4.38 (m, 1H), 4.28-4.19 (m, 3H), 3.25-3.21 (m, 1H), 3.03-2.98 (m, 1H), 2.95-2.68 (m, 1H), 2.66-2.60 (m, 1H), 2.05-2.03 (m, 1H), 1.99-1.91 (m, 1H), 1.48 (s, 9H), 1.32-1.28 (m, 3H).
Step a. To a solution of Intermediate B47a (0.8 g, 2.83 mmol) in THF (8 mL) and MeOH (1 mL) was added LiBH4 (185 mg, 8.5 mmol) and the mixture was stirred at 10° C. for 12 h. Upon completion, the mixture was quenched with sat. aq. NH4Cl (0.2 mL), diluted with EtOAc (30 mL), filtered and evaporated. The residue was purified by column chromatography (PE/EtOAc=1/3) to give tert-butyl (rac-cis)-3-cyano-4-(hydroxymethyl)piperidine-1-carboxylate (500 mg, 73% yield) as a colourless oil.
1H NMR (400 MHz, CDCl3) δ ppm 4.50-4.36 (m, 1H), 4.35-4.20 (m, 1H), 3.67-3.53 (m, 2H), 3.06-3.00 (m, 1H), 2.92-2.73 (m, 1H), 2.72-2.54 (m, 1H), 1.89-1.86 (m, 1H), 1.56-1.52 (m, 3H), 1.42 (s, 9H).
Step b. To a solution of tert-butyl (rac-cis)-3-cyano-4-(hydroxymethyl)piperidine-1-carboxylate (450 mg, 1.87 mmol) in DCM (10 mL) was added Dess-Martin periodinane (1.59 g, 3.75 mmol) and the mixture was stirred at 15° C. for 1 h. Upon completion, the mixture was filtered, evaporated and purified by column chromatography (PE/EtOAc=1/1) to give the title compound (434 mg, crude) as a yellow oil which was used directly in subsequent steps.
The title compound was prepared in a similar manner to Intermediate B48a, using Intermediate B47b.
1H NMR (400 MHz, CDCl3) δ ppm 9.74 (s, 1H), 4.51-4.13 (m, 1H), 4.12-3.87 (m, 1H), 3.81-3.56 (m, 1H), 3.52-3.10 (m, 1H), 3.07-2.90 (m, 1H), 2.88-2.77 (m, 1H), 2.23-2.12 (m, 1H), 1.72-1.54 (m, 1H), 1.48 (s, 9H).
Step a. To a solution of 2,3-dimethylpyrazine (10 g, 92.5 mmol) in CCl4 (320 mL) was added NBS (41.2 g, 231 mmol) and AIBN (106 mg, 0.65 mmol). The mixture was stirred at 80° C. for 1 h under a N2 atmosphere. After 1 h, additional AIBN (106 mg, 0.65 mmol) was added and stirred at 80° C. for a further 5 h under a N2 atmosphere. Upon completion, the mixture was filtered and the filter cake was washed with CCl4 (100 mL). The filtrate was evaporated and purified by column chromatography (PE/EtOAc=20/1) to give 2,3-bis(bromomethyl)pyrazine (20 g, 31% yield) as a purple oil.
m/z ES+ [M+H]+ 266.8; 1H NMR (400 MHz, CDCl3) δ ppm 8.51 (s, 2H), 4.72 (s, 4H).
Step b. To a solution of 2,3-bis(bromomethyl)pyrazine (40 g, 116 mmol) in DMF (1500 mL) was added a solution of triphenylmethanamine (90.1 g, 347 mmol) in DMF (500 mL) at 0° C. The mixture was stirred at 20° C. for 1 h followed by 60° C. for 12 h. Upon completion, the mixture was evaporated, diluted with water (400 mL) and extracted with EtOAc (3×400 mL). The combined organic layers were washed with brine (2×500 mL), dried over Na2SO4, evaporated and purified by column chromatography (PE/EtOAc=7/1) followed by trituration with MeOH (100 mL) at 20° C. to give 6-trityl-6,7-dihydro-5H-pyrrolo[3,4-b]pyrazine (16 g, 37% yield) as a yellow solid.
m/z ES+ [M+H]+ 364.0; 1H NMR (400 MHz, CDCl3) δ ppm 8.22 (s, 2H), 7.64-7.56 (m, 6H), 7.35-7.28 (m, 6H), 7.23-7.16 (m, 3H), 4.08 (s, 4H).
Step c. A solution of 6-trityl-6,7-dihydro-5H-pyrrolo[3,4-b]pyrazine (16.0 g, 44.0 mmol) in 4 M HCl in MeOH (120 mL) was stirred at 20° C. for 2 h. Upon completion, the mixture was evaporated, diluted with water (200 mL) and extracted with EtOAc (3×150 mL). The aqueous phase was lyophilized to give 6,7-dihydro-5H-pyrrolo[3,4-b]pyrazine hydrochloride (7 g) as a black solid.
1H NMR (400 MHz, DMSO-d6) δ ppm 10.60 (br s, 2H), 8.60 (s, 2H), 4.58-4.48 (m, 4H).
Step d. To a solution of 6,7-dihydro-5H-pyrrolo[3,4-b]pyrazine hydrochloride (8.67 g, 55.0 mmol) in THF (200 mL) and water (100 mL) was added Na2CO3 (14.6 g, 138 mmol) and CbzCl (11.3 g, 66.0 mmol). The reaction mixture was stirred at 20° C. for 5 h. Upon completion, the mixture was diluted with water (100 mL) and extracted with EtOAc (3×200 mL). The combined organic layers were washed with brine (500 mL), dried over Na2SO4, evaporated and purified by column chromatography (PE/EtOAc=2/1) to give benzyl 5,7-dihydro-6H-pyrrolo[3,4-b]pyrazine-6-carboxylate (7.5 g, 64% yield) as a yellow solid.
m/z ES+ [M+H]+ 256.1; 1H NMR (400 MHz, CDCl3) δ ppm 8.46 (s, 2H), 7.45-7.33 (m, 5H), 5.26 (s, 2H), 4.85-4.83 (m, 4H).
Step e. To a solution of benzyl 5,7-dihydro-6H-pyrrolo[3,4-b]pyrazine-6-carboxylate (10 g, 39.2 mmol) in DCM (150 mL) was added m-CPBA (80%; 9.3 g, 43.1 mmol). The reaction mixture was stirred at 25° C. for 12 h. Upon completion, the mixture was diluted with sat. aq. Na2S2O3 (150 mL) and extracted with DCM (200 mL). The combined organic layers were washed with sat. aq. Na2S2O3 (150 mL), brine (300 mL), dried over Na2SO4, evaporated and purified by column chromatography (PE/EtOAc=0/1) to give 6-((benzyloxy)carbonyl)-6,7-dihydro-5H-pyrrolo[3,4-b]pyrazine 1-oxide (8.3 g, 76% yield) as a yellow solid.
m/z ES+ [M+H]+ 272.0; 1H NMR (400 MHz, CDCl3) δ ppm 8.39-8.36 (m, 1H), 8.01 (d, J=4.0 Hz, 1H), 7.45-7.32 (m, 5H), 5.24 (d, J=3.2 Hz, 2H), 4.90 (s, 2H), 4.89-4.84 (m, 2H).
Step f. To a solution of 6-((benzyloxy)carbonyl)-6,7-dihydro-5H-pyrrolo[3,4-b]pyrazine 1-oxide (14.3 g, 52.7 mmol) in DMF (200 mL) was added oxalyl chloride (16.7 g, 132 mmol) dropwise at 0° C. The reaction mixture was stirred at 40° C. for 12 h. Upon completion, the mixture was basified to pH 8 with sat. aq. NaHCO3, diluted with water (500 mL) and extracted with EtOAc (3×500 mL). The combined organic layers were washed with brine (2×800 mL), dried over Na2SO4, evaporated and purified by column chromatography (PE/EtOAc=7/1) to give benzyl 2-chloro-5,7-dihydro-6H-pyrrolo[3,4-b]pyrazine-6-carboxylate (11.6 g, 76% yield) as a yellow solid.
m/z ES+ [M+H]+ 290.1; 1H NMR (400 MHz, CDCl3) δ ppm 8.48 (d, J=4.0 Hz, 1H), 7.49-7.32 (m, 5H), 5.25 (s, 2H), 4.84-4.77 (m, 4H).
Step g. A mixture of benzyl 2-chloro-5,7-dihydro-6H-pyrrolo[3,4-b]pyrazine-6-carboxylate (10 g, 34.5 mmol), potassium trifluoro(vinyl)borate (CAS: 13682-77-4; 9.25 g, 69.0 mmol), Pd(dppf)Cl2 (2.53 g, 3.45 mmol) and Na2CO3 (9.15 g, 86.3 mmol) in 1,4-dioxane (200 mL) and water (30 mL) was degassed and purged with N2 3 times. The reaction mixture was stirred at 80° C. for 12 h under a N2 atmosphere. Upon completion, the mixture was diluted with water (250 mL) and extracted with EtOAc (3×250 mL). The combined organic layers were washed with brine (600 mL), dried over Na2SO4, evaporated and purified by column chromatography (PE/EtOAc=9/1) to give benzyl 2-vinyl-5,7-dihydro-6H-pyrrolo[3,4-b]pyrazine-6-carboxylate (9.04 g, 93% yield) as an off-white solid.
m/z ES+ [M+H]+ 282.2; 1H NMR (400 MHz, CDCl3) δ ppm 8.43 (s, 1H), 7.46-7.31 (m, 5H), 6.82-6.79 (m, 1H), 6.41-6.34 (m, 1H), 5.63-5.69 (m, 1H), 5.26 (s, 2H), 4.81 (s, 4H).
Step h. To a solution of benzyl 2-vinyl-5,7-dihydro-6H-pyrrolo[3,4-b]pyrazine-6-carboxylate (8.8 g, 31.3 mmol) in THF (150 mL) and water (150 mL) was added NaIO4 (20.1 g, 93.9 mmol) and OsO4 (795 mg, 3.13 mmol) at 0° C. The reaction mixture was stirred at 0° C. for 1 h. Upon completion, the mixture was quenched with sat. aq. Na2S2O3 (200 mL) and stirred at 25° C. for 10 min. The mixture was diluted with water (50 mL) and extracted with EtOAc (3×250 mL). The combined organic layers were washed with sat. aq. Na2S2O3 (400 mL), brine (500 mL), dried over Na2SO4, evaporated and purified by column chromatography (PE/EtOAc=3/1) to give the title compound (6.4 g, 72% yield) as a yellow solid.
m/z ES+ [M+H]+ 284.0; 1H NMR (400 MHz, CDCl3) δ ppm 10.14 (d, J=6.4 Hz, 1H), 9.06 (d, J=4.0 Hz, 1H), 7.48-7.32 (m, 5H), 5.27 (s, 2H), 4.92 (d, J=3.2 Hz, 4H).
Step a. To a solution of (tert-butoxycarbonyl)serine (50 g, 244 mmol) and imidazole (33.2 g, 487 mmol) in DMF (500 mL) was added TBSCl (36.7 g, 244 mmol) at 0° C. The reaction mixture was stirred at 20° C. for 12 h. Upon completion, the mixture was diluted with water (1.5 L) and extracted with EtOAc (3×1 L). The combined organic layers were washed with brine (3×1 L), dried over Na2SO4 and evaporated to give N-(tert-butoxycarbonyl)-O-(tert-butyldimethylsilyl)serine (64 g, 82% yield) as a colourless oil.
1H NMR (400 MHz, CDCl3) δ ppm 5.49-5.30 (m, 1H), 4.37-4.21 (m, 1H), 4.10-4.02 (m, 1H), 3.89-3.81 (m, 1H), 1.46 (s, 9H), 0.89 (s, 9H), −0.06 (s, 6H).
Step b. A solution of N-(tert-butoxycarbonyl)-O-(tert-butyldimethylsilyl)serine (66.9 g, 209 mmol) and CDI (40.8 g, 251 mmol) in THF (670 mL) was stirred at 20° C. for 1 h, after which potassium 3-methoxy-3-oxopropanoate (32.7 g, 209 mmol) and MgCl2 (19.9 g, 209 mmol) was added. The reaction mixture was stirred at 50° C. for a further 16 h. Upon completion, the mixture was diluted with water (500 mL) and extracted with EtOAc (500 mL). The organic layer was washed with brine (500 mL), dried over Na2SO4, evaporated and purified by column chromatography (PE/EtOAc=10/1) to give methyl 4-((tert-butoxycarbonyl)amino)-5-((tert-butyldimethylsilyl)oxy)-3-oxopentanoate (27.6 g, 35% yield) as a colourless oil.
1H NMR (400 MHz, CDCl3) δ ppm 5.49-5.39 (m, 1H), 4.45-4.36 (m, 1H), 4.12-4.04 (m, 1H), 3.83-3.79 (m, 1H), 3.74 (s, 3H), 3.62 (d, J=3.6 Hz, 2H), 1.46 (s, 9H), 0.88 (s, 9H), 0.06 (s, 6H).
Step c. To a solution of methyl 4-((tert-butoxycarbonyl)amino)-5-((tert-butyldimethylsilyl)oxy)-3-oxopentanoate (27.6 g, 73.5 mmol) in THF (280 mL) was added t-BuOK (9.07 g, 80.9 mmol) at 0° C. The mixture was stirred at 0° C. for 45 min, after which DABCO (9.07 g, 80.9 mmol) and (Z)—N-(2-chloro-3-(dimethylamino)allylidene)-N-methylmethanaminium hexafluorophosphate (23.7 g, 77.2 mmol) was added at 0° C. The mixture was stirred at 20° C. for 3 h and then ammonium acetate (6.23 g, 80.9 mmol) was added. The reaction mixture was stirred at 20° C. for a further 16 h. Upon completion, the mixture was diluted with water (500 mL) and extracted with EtOAc (3×300 mL). The combined organic layers were washed with brine (300 mL), dried over Na2SO4, evaporated and purified by column chromatography (PE/EtOAc=10/1) to give methyl 5-chloro-2-(2,2,3,3,10,10-hexamethyl-8-oxo-4,9-dioxa-7-aza-3-silaundecan-6-yl)nicotinate (23 g, 69% yield) as a white solid.
m/z ES+ [M+H]+ 445.1; 1H NMR (400 MHz, CDCl3) δ ppm 8.63 (d, J=2.4 Hz, 1H), 8.19 (d, J=2.4 Hz, 1H), 6.01-5.78 (m, 2H), 3.94 (s, 3H), 3.90-3.83 (m, 1H), 3.82-3.74 (m, 1H), 1.44 (s, 9H), 0.77 (s, 9H), −0.09 (s, 3H), −0.11 (s, 3H).
Step d. To a solution of methyl 5-chloro-2-(2,2,3,3,10,10-hexamethyl-8-oxo-4,9-dioxa-7-aza-3-silaundecan-6-yl)nicotinate (13 g, 29.2 mmol) in EtOH (130 mL) was added NaBH4 (2.21 g, 58.4 mmol) and CaCl2 (3.24 g, 29.2 mmol) at 0° C. The reaction mixture was stirred at 0° C. for 2 h. Upon completion, the mixture was quenched with sat. aq. NH4Cl (300 mL) and extracted with EtOAc (3×300 mL). The combined organic layers were washed with brine (300 mL), dried over Na2SO4, evaporated and purified by column chromatography (PE/EtOAc=5/1) to give tert-butyl (2-((tert-butyldimethylsilyl)oxy)-1-(5-chloro-3-(hydroxymethyl)pyridin-2-yl)ethyl)carbamate (6.6 g, 53% yield) as a colourless oil.
m/z ES+ [M+H]+ 417.3; 1H NMR (400 MHz, CDCl3) δ ppm 8.48 (d, J=2.4 Hz, 1H), 7.69 (d, J=2.4 Hz, 1H), 5.57 (d, J=8.0 Hz, 1H), 5.18-5.06 (m, 1H), 4.80-4.66 (m, 2H), 4.02-3.95 (m, 1H), 3.75-3.62 (m, 2H), 1.42 (s, 9H), 0.75 (s, 9H), −0.05 (s, 3H), −0.11 (s, 3H).
Step e. To a solution of tert-butyl (2-((tert-butyldimethylsilyl)oxy)-1-(5-chloro-3-(hydroxymethyl)pyridin-2-yl)ethyl)carbamate (5.6 g, 13.4 mmol) in DCM (60 mL) was added TEA (4.08 g, 40.3 mmol) and TsCl (5.12 g, 26.9 mmol) at 0° C. The reaction mixture was stirred at 0° C. for 1 h. Upon completion, the mixture was quenched with sat. aq. NaHCO3 (100 mL) and extracted with EtOAc (100 mL). The organic layer was washed with brine (100 mL), dried over Na2SO4, evaporated and purified by column chromatography (PE/EtOAc=10/1) to give (5-chloro-2-(2,2,3,3,10,10-hexamethyl-8-oxo-4,9-dioxa-7-aza-3-silaundecan-6-yl)pyridin-3-yl)methyl 4-methylbenzenesulfonate (5.3 g, 69% yield) as a colourless oil.
m/z ES+ [M+H]+ 571.1; 1H NMR (400 MHz, CDCl3) δ ppm 8.45 (d, J=2.4 Hz, 1H), 7.84 (d, J=8.0 Hz, 2H), 7.69 (d, J=2.4 Hz, 1H), 7.36 (d, J=8.0 Hz, 2H), 5.68 (d, J=7.2 Hz, 1H), 5.31-5.16 (m, 2H), 4.87-4.80 (m, 1H), 3.93-3.84 (m, 1H), 3.52-3.44 (m, 1H), 2.46 (s, 3H), 1.60 (s, 9H), 1.43 (s, 9H), −0.16-−0.13 (m, 3H), −0.20 (s, 3H).
Step f. To a solution of (5-chloro-2-(2,2,3,3,10,10-hexamethyl-8-oxo-4,9-dioxa-7-aza-3-silaundecan-6-yl)pyridin-3-yl)methyl 4-methylbenzenesulfonate (4.8 g, 8.40 mmol) in DMF (150 mL) was added NaH (1.01 g, 25.2 mmol, 60% dispersion in mineral oil) at 0° C. The reaction mixture was stirred at 0° C. for 1 h. Upon completion, the mixture was diluted with water (500 mL) and extracted with EtOAc (500 mL). The organic layer was washed with brine (500 mL), dried over Na2SO4, evaporated and purified by reverse phase flash chromatography (water (0.1% NH4OH)/MeCN) to give tert-butyl 7-(((tert-butyldimethylsilyl)oxy)methyl)-3-chloro-5,7-dihydro-6H-pyrrolo[3,4-b]pyridine-6-carboxylate (2.2 g, 66% yield) as a colourless oil.
m/z ES+ [M+H]+ 399.0; 1H NMR (400 MHz, CDCl3) δ ppm 8.46 (s, 1H), 7.57-7.49 (m, 1H), 4.99-4.83 (m, 1H), 4.80-4.63 (m, 1H), 4.60-4.50 (m, 1H), 4.44-4.18 (m, 1H), 4.07-3.98 (m, 1H), 1.53 (s, 9H), 0.68 (d, J=2.4 Hz, 9H), −0.07 (d, J=4.0 Hz, 3H), −0.18 (d, J=8.4 Hz, 3H).
Step g. This step was conducted in a similar manner to Intermediate B49, step g, and an additional chiral SFC step was conducted to afford the title compounds.
Intermediate B50a:
m/z ES+ [M+H]+ 391.3; 1H NMR (400 MHz, CDCl3) δ ppm 8.50-8.45 (m, 1H), 7.66-7.54 (m, 1H), 6.79-6.68 (m, 1H), 5.86-5.76 (m, 1H), 5.42-5.33 (m, 1H), 5.00-4.83 (m, 1H), 4.81-4.65 (m, 1H), 4.62-4.51 (m, 1H), 4.20-4.45 (m, 1H), 4.10-4.00 (m, 1H), 1.53 (s, 9H), 0.67 (d, J=3.2 Hz, 9H), −0.08 (d, J=5.6 Hz, 3H), −0.20 (d, J=10.4 Hz, 3H).
Intermediate B50b:
m/z ES+ [M+H]+ 391.3; 1H NMR (400 MHz, CDCl3) δ ppm 8.48 (d, J=2.8 Hz, 1H), 7.67-7.54 (m, 1H), 6.65-6.80 (m, 1H), 5.75-5.90 (m, 1H), 5.37 (d, J=11.2 Hz, 1H), 5.02-4.84 (m, 1H), 4.82-4.64 (m, 1H), 4.61-4.49 (m, 1H), 4.46-4.16 (m, 1H), 4.12-3.98 (m, 1H), 1.53 (s, 9H), 0.67 (d, J=3.2 Hz, 9H), −0.08 (d, J=5.6 Hz, 3H), −0.20 (d, J=10.4 Hz, 3H).
Step a. This step was conducted in a similar manner to Intermediate B49, step h, using Intermediate B50a.
m/z ES+ [M+H]+ 393.4; 1H NMR (400 MHz, CDCl3) δ ppm 10.12 (s, 1H), 8.96 (s, 1H), 8.08-7.95 (m, 1H), 5.11-4.92 (m, 1H), 4.90-4.71 (m, 1H), 4.68-4.56 (m, 1H), 4.50-4.21 (m, 1H), 4.00-4.18 (m, 1H), 1.54 (s, 9H), 0.65 (d, J=2.8 Hz, 9H), −0.07 (d, J=4.0 Hz, 3H), −0.19 (d, J=8.4 Hz, 3H).
The title compound was prepared in a similar manner to Intermediate B51a, using Intermediate B50b.
m/z ES+ [M+H]+ 393.4; 1H NMR (400 MHz, CDCl3) δ ppm 10.12 (s, 1H), 8.96 (s, 1H), 8.09-7.97 (m, 1H), 5.10-4.93 (m, 1H), 4.90-4.73 (m, 1H), 4.68-4.58 (m, 1H), 4.49-4.21 (m, 1H), 4.14-4.07 (m, 1H), 1.54 (s, 9H), 0.65 (d, J=2.8 Hz, 9H), −0.07 (d, J=4.4 Hz, 3H), −0.19 (d, J=8.8 Hz, 3H).
Step a. To a solution of methyl 5-bromopicolinate (15 g, 69.4 mmol) in DCM (250 mL) was added m-CPBA (80%; 30.0 g, 139 mmol). The reaction mixture was stirred at 25° C. for 12 h. Upon completion, the mixture was diluted with sat. aq. Na2SO3 (500 mL) and extracted with DCM (500 mL). The combined organic layers were washed with sat. aq. Na2SO3 (300 mL), brine (500 mL), dried over Na2SO4, evaporated and purified by column chromatography (PE/EtOAc=1/2) to give 5-bromo-2-(methoxycarbonyl)pyridine 1-oxide (11.7 g, 70% yield) as an off-white solid.
m/z ES+ [M+H]+ 234.0; 1H NMR (400 MHz, CDCl3) δ ppm 8.41 (s, 1H), 7.55 (d, J=8.4 Hz, 1H), 7.41 (d, J=8.4 Hz, 1H), 4.00 (s, 3H).
Step b. A mixture of 5-bromo-2-(methoxycarbonyl)pyridine 1-oxide (11.7 g, 50.4 mmol), TMSCN (50.0 g, 504 mmol) and dimethylcarbamoyl chloride (54.2 g, 504 mmol) was stirred at 50° C. for 16 h under a N2 atmosphere. Upon completion, the mixture was basified to pH 8 with 1 M NaOH and extracted with EtOAc (3×250 mL). The combined organic layers were washed with brine (600 mL), dried over Na2SO4, evaporated and purified by column chromatography (PE/EtOAc=7/1) to give methyl 5-bromo-6-cyanopicolinate (10.4 g, 85% yield) as an off-white solid.
m/z ES+ [M+H]+ 241.0; 1H NMR (400 MHz, CDCl3) δ ppm 8.24-8.15 (m, 2H), 4.04 (s, 3H).
Step c. To a solution of methyl 5-bromo-6-cyanopicolinate (10.4 g, 43.2 mmol) in THF (120 mL) was added a solution of borane tetrahydrofuran complex (1 M in THF, 215.73 mL) dropwise at 0° C. The reaction mixture was stirred at 20° C. for 3 h. Upon completion, the mixture was quenched with MeOH (50 mL) at 0° C. and evaporated to give methyl 6-(aminomethyl)-5-bromopicolinate (12.0 g, crude) as a yellow solid.
Step d. To a solution of methyl 6-(aminomethyl)-5-bromopicolinate (12.0 g, 42.6 mmol) in THF (150 mL) was added NaHCO3 (7.16 g, 85.3 mmol) and Boc2O (11.2 g, 51.2 mmol). The reaction mixture was stirred at 20° C. for 12 h. Upon completion, the mixture was diluted with water (150 mL) and extracted with EtOAc (3×150 mL). The combined organic layers were washed with brine (300 mL), dried over Na2SO4, evaporated and purified by column chromatography (PE/EtOAc=7/1) to give methyl 5-bromo-6-(((tert-butoxycarbonyl)amino)methyl)picolinate (4.5 g, 29% yield) as a yellow solid.
m/z ES+ [M+Na]+ 367.0; 1H NMR (400 MHz, CDCl3) δ ppm 8.01 (d, J=8.0 Hz, 1H), 7.90 (d, J=8.4 Hz, 1H), 5.96 (s, 1H), 4.60 (s, 2H), 4.00 (s, 3H), 1.50 (s, 9H).
Step e. A mixture of methyl 5-bromo-6-(((tert-butoxycarbonyl)amino)methyl)picolinate (2.5 g, 7.24 mmol), potassium trifluoro(vinyl)borate (CAS: 13682-77-4; 1.94 g, 14.5 mmol), Pd(dppf)Cl2 (265 mg, 0.36 mmol) and Na2CO3 (1.92 g, 18.1 mmol) in 1,4-dioxane (40 mL) and water (4 mL) was degassed and purged with N2 3 times. The reaction mixture was stirred at 80° C. for 24 h under a N2 atmosphere. Upon completion, the mixture was diluted with water (50 mL) and extracted with EtOAc (3×50 mL). The combined organic layers were washed with brine (150 mL), dried over Na2SO4, evaporated and purified by column chromatography (PE/EtOAc=8/1) to give methyl 6-(((tert-butoxycarbonyl)amino)methyl)-5-vinylpicolinate (1.68 g, 76% yield) as a yellow solid.
m/z ES+ [M+Na]+ 315.0; 1H NMR (400 MHz, CDCl3) δ ppm 8.03 (d, J=8.0 Hz, 1H), 7.92 (d, J=8.0 Hz, 1H), 6.99-6.92 (m, 1H), 5.95 (br s, 1H), 5.83 (d, J=17.2 Hz, 1H), 5.59 (d, J=11.2 Hz, 1H), 4.58 (s, 2H), 4.01 (s, 3H), 1.49 (s, 9H).
Step f. To a solution of methyl 6-(((tert-butoxycarbonyl)amino)methyl)-5-vinylpicolinate (1.7 g, 5.82 mmol) in MeCN (15 mL), tBuOH (15 mL) and water (15 mL) was added OsO4 (148 mg, 0.58 mmol) and NMO (818 mg, 6.98 mmol). The reaction mixture was stirred at 20° C. for 2 h. Upon completion, the mixture was quenched with sat. aq. Na2SO3 (25 mL) at 20° C. and extracted with EtOAc (3×25 mL). The combined organic layers were washed with sat. aq. Na2SO3 (35 mL), brine (60 mL), dried over Na2SO4, evaporated and purified by column chromatography (PE/EtOAc=1/2) to give methyl 6-(((tert-butoxycarbonyl)amino)methyl)-5-(1,2-dihydroxyethyl)picolinate (1.5 g, 78% yield) as a yellow solid.
m/z ES+ [M+H]+ 327.2; 1H NMR (400 MHz, CDCl3) δ ppm 8.10-8.01 (m, 2H), 6.04 (s, 1H), 5.26-5.17 (m, 1H), 4.69-4.57 (m, 1H), 4.53-4.43 (m, 1H), 4.15-4.03 (m, 1H), 3.99 (s, 3H), 3.84-3.81 (m, 1H), 3.70-3.67 (m, 1H), 3.00 (br s, 1H), 1.44 (s, 9H).
Step g. This step was conducted in a similar manner to Intermediate B50a and Intermediate B50b, step a, using TBDPSCl.
m/z ES+ [M+H]+ 565.3; 1H NMR (400 MHz, CDCl3) δ ppm 8.05-8.01 (m, 2H), 8.00-7.96 (m, 2H), 7.52-7.49 (m, 2H), 7.46-7.41 (m, 6H), 5.84 (s, 1H), 5.14 (s, 1H), 4.52-4.51 (m, 1H), 4.26-4.25 (m, 1H), 3.89 (s, 3H), 3.85-3.82 (m, 1H), 3.70-3.66 (m, 1H), 3.31 (s, 1H), 1.44 (s, 9H), 1.05 (s, 9H).
Step h. To a solution of methyl 6-(((tert-butoxycarbonyl)amino)methyl)-5-(2-((tert-butyldiphenylsilyl)oxy)-1-hydroxyethyl)picolinate (2 g, 3.54 mmol) in DCM (50 mL) was added TEA (1.08 g, 10.6 mmol) and methanesulfonic anhydride (925 mg, 5.31 mmol). The reaction mixture was stirred at 20° C. for 2 h. Upon completion, the mixture was diluted with water (50 mL) and extracted with DCM (2×50 mL). The combined organic layers were washed with brine (120 mL), dried over Na2SO4, evaporated and purified by column chromatography (PE/EtOAc=4/1) to give methyl 6-(((tert-butoxycarbonyl)amino)methyl)-5-(2-((tert-butyldiphenylsilyl)oxy)-1-((methylsulfonyl)oxy)ethyl)picolinate (2 g, 88% yield) as a white solid.
m/z ES+ [M+H]+ 643.2; 1H NMR (400 MHz, CDCl3) δ ppm 8.06-8.00 (m, 1H), 7.99-7.92 (m, 1H), 7.60-7.51 (m, 4H), 7.48-7.41 (m, 2H), 7.41-7.34 (m, 4H), 6.08 (s, 1H), 5.68 (t, J=4.8 Hz, 1H), 4.62-4.57 (m, 1H), 4.39-4.35 (m, 1H), 4.01 (s, 3H), 3.99-3.93 (m, 1H), 3.89-3.83 (m, 1H), 3.02 (s, 3H), 1.45 (s, 9H), 1.02 (s, 9H).
Step i. To a solution of methyl 6-(((tert-butoxycarbonyl)amino)methyl)-5-(2-((tert-butyldiphenylsilyl)oxy)-1-((methylsulfonyl)oxy)ethyl)picolinate (150 mg, 0.23 mmol) in NMP (5 mL) was added K3PO4 (149 mg, 0.70 mmol). The reaction mixture was stirred at 60° C. for 48 h. Upon completion, the mixture was diluted with water (5 mL) and extracted with EtOAc (3×5 mL). The combined organic layers were washed with brine (2×15 mL), dried over Na2SO4, evaporated and purified by prep-TLC (PE/EtOAc=1/1) to give 6-(tert-butyl) 2-methyl 5-(((tert-butyldiphenylsilyl)oxy)methyl)-5,7-dihydro-6H-pyrrolo[3,4-b]pyridine-2,6-dicarboxylate (30 mg, 23% yield) as a brown solid.
m/z ES+ [M+H]+ 547.3; 1H NMR (400 MHz, CDCl3) δ ppm 8.14-8.04 (m, 1H), 7.72-7.65 (m, 1H), 7.53-7.47 (m, 3H), 7.44-7.38 (m, 3H), 7.38-7.31 (m, 4H), 5.23-5.08 (m, 1H), 4.97-4.82 (m, 1H), 4.80-4.72 (m, 1H), 4.05 (s, 3H), 4.02-3.95 (m, 1H), 3.95-3.87 (m, 1H), 1.54-1.41 (m, 9H), 0.90 (d, J=12.8 Hz, 9H).
Step j. To a solution of 6-(tert-butyl) 2-methyl 5-(((tert-butyldiphenylsilyl)oxy)methyl)-5,7-dihydro-6H-pyrrolo[3,4-b]pyridine-2,6-dicarboxylate (320 mg, 0.59 mmol) in THF (5 mL) and MeOH (0.5 mL) was added LiBH4 (64 mg, 2.93 mmol). The reaction mixture was stirred at 20° C. for 2 h. Upon completion, the mixture was quenched with sat. aq. NH4Cl (10 mL) at 20° C. and extracted with EtOAc (3×10 mL). The combined organic layers were washed with brine (25 mL), dried over Na2SO4, evaporated and purified by column chromatography (PE/EtOAc=6/1) to give tert-butyl 5-(((tert-butyldiphenylsilyl)oxy)methyl)-2-(hydroxymethyl)-5,7-dihydro-6H-pyrrolo[3,4-b]pyridine-6-carboxylate (210 mg, 66% yield) as a yellow solid.
m/z ES+ [M+H]+ 519.3; 1H NMR (400 MHz, CDCl3) δ ppm 7.68-7.48 (m, 4H), 7.46-7.29 (m, 7H), 7.20-7.15 (m, 1H), 5.19-5.04 (m, 1H), 4.87-4.74 (m, 3H), 4.71-4.61 (m, 1H), 4.08-3.81 (m, 2H), 3.52-3.39 (m, 1H), 1.55-1.41 (m, 9H), 0.91 (d, J=13.2 Hz, 9H).
Step k. To a solution of tert-butyl 5-(((tert-butyldiphenylsilyl)oxy)methyl)-2-(hydroxymethyl)-5,7-dihydro-6H-pyrrolo[3,4-b]pyridine-6-carboxylate (210 mg, 0.41 mmol) in DCM (5 mL) was added MnO2 (704 mg, 8.10 mmol). The reaction mixture was stirred at 20° C. for 12 h. Upon completion, the mixture was filtered, evaporated and purified by Prep-TLC (PE/EtOAc=2/1) to give the title compound (120 mg, 57% yield) as a white solid.
m/z ES+ [M+H]+ 517.2; 1H NMR (400 MHz, CDCl3) δ ppm 10.10 (d, J=6.8 Hz, 1H), 7.92-7.90 (m, 1H), 7.81-7.67 (m, 1H), 7.55-7.43 (m, 4H), 7.41-7.30 (m, 6H), 5.25-5.09 (m, 1H), 4.96-4.75 (m, 2H), 4.01-3.90 (m, 2H), 1.56 (d, J=6.8 Hz, 9H), 0.89 (d, J=10.8 Hz, 9H).
Step a. To a solution of 2-chloro-6,7-dihydro-5H-pyrrolo[3,4-b]pyridine hydrochloride (CAS: 1841081-37-5; 24.0 g, 125 mmol) in 1,4-dioxane (240 mL) was added Boc2O (41.1 g, 188 mmol), TEA (38.1 g, 376 mmol) and DMAP (1.53 g, 12.5 mmol). The reaction mixture was stirred at 25° C. for 30 min. Upon completion, the mixture was diluted with water (250 mL) and extracted with EtOAc (2×150 mL). The combined organic layers were washed with brine (200 mL), dried over Na2SO4, evaporated and purified by column chromatography (PE/EtOAc=3/1) to give tert-butyl 2-chloro-5,7-dihydro-6H-pyrrolo[3,4-b]pyridine-6-carboxylate (30.0 g, 93% yield) as a white solid.
m/z ES+ [M+H]+ 255.0; 1H NMR (400 MHz, DMSO-d6) δ ppm 7.82 (t, J=7.2 Hz, 1H), 7.42 (d, J=8.0 Hz, 1H), 4.47-4.56 (m, 4H), 1.46 (s, 9H).
Step b. To a solution of tert-butyl 2-chloro-5,7-dihydro-6H-pyrrolo[3,4-b]pyridine-6-carboxylate (30.0 g, 117 mmol) in DMSO (150 mL) and MeOH (150 mL) was added Pd(OAc)2 (5.29 g, 23.5 mmol), DPPP (9.72 g, 23.5 mmol) and TEA (119 g, 1.18 mol). The reaction mixture was stirred at 60° C. for 12 h under a CO atmosphere (50 psi). Upon completion, the mixture was filtered, evaporated and purified by column chromatography (PE/EtOAc=2/1) to give 6-(tert-butyl) 2-methyl 5,7-dihydro-6H-pyrrolo[3,4-b]pyridine-2,6-dicarboxylate (32.0 g, 96% yield) as a yellow solid.
m/z ES+ [M+H]+ 279.1; 1H NMR (400 MHz, DMSO-d6) δ ppm 7.89-8.01 (m, 2H), 4.67 (d, J=11.6 Hz, 2H), 4.59 (d, J=6.4 Hz, 2H), 3.88 (s, 3H), 1.46 (s, 9H).
Step c. To a solution of 6-(tert-butyl) 2-methyl 5,7-dihydro-6H-pyrrolo[3,4-b]pyridine-2,6-dicarboxylate (43.5 g, 156 mmol) in THF (545 mL) at −78° C. was slowly added DIBAL-H (1 M in toluene, 390 mL) over 30 min. The reaction mixture was stirred at −78° C. for 3 h. Upon completion, the mixture was added dropwise to MeOH (700 mL) at −40° C. under a N2 atmosphere. The mixture was stirred for 30 min, after which 0.5 M HCl (800 mL) was added and the mixture was stirred at 15° C. for a further 30 min. The aqueous mixture was extracted with EtOAc (2×500 mL) and the combined organic layers were dried over Na2SO4, evaporated and purified by column chromatography (PE/EtOAc=5/1) to give the title compound (25.0 g, 64% yield) as a white solid.
m/z ES+ [M+H]+ 249.2; 1H NMR (400 MHz, DMSO-d6) δ ppm 9.97 (d, J=2.0 Hz, 1H), 7.96-8.04 (m, 1H), 7.87 (d, J=7.6 Hz, 1H), 4.63-4.76 (m, 4H), 1.47 (s, 9H).
Step a. To a solution of 6-chloronicotinic acid (CAS: 5326-23-8; 23.0 g, 145 mmol) in toluene (200 mL) was added DMF (872 mg, 11.9 mmol) at 0° C., after which SOCl2 (86.8 g, 729 mmol) was added to the mixture slowly, keeping the temperature between 0-5° C. The reaction mixture was stirred at 0° C. for 30 min followed by 85° C. for a further 5 h. Upon completion, the mixture was evaporated to give 6-chloronicotinoyl chloride (25 g, crude) as yellow oil. To a solution of the acid chloride (25.0 g, 142.1 mmol) in DCM (250 mL) was added diisopropylamine (57.5 g, 568 mmol). The mixture was stirred at 0° C. for 2 h. Upon completion, the reaction mixture was quenched with water (50 mL) at 0° C. and extracted with DCM (3×50 mL). The combined organic layers were washed with brine (60 mL), dried over Na2SO4 and evaporated to give 6-chloro-N,N-diisopropylnicotinamide (32.0 g, 93% yield) as a yellow solid.
1H NMR (400 MHz, CDCl3) δ ppm 8.37 (d, J=2.0 Hz, 1H), 7.72-7.54 (m, 1H), 7.37 (d, J=8.4 Hz, 1H), 3.89-3.45 (m, 2H), 1.58-1.11 (m, 12H).
Step b. A solution of LDA (2 M in THF, 93.4 mL) was added dropwise to a solution of 6-chloro-N,N-diisopropylnicotinamide (30.0 g, 124 mmol) in THF (300 mL) at −78° C. under a N2 atmosphere. The mixture was stirred at −78° C. for 2 h, after which DMF (23.3 g, 318 mmol) was added slowly. The reaction mixture was stirred at −78° C. for 1 h, slowly warmed to 25° C. and stirred at that temperature for an additional 30 min. Upon completion, the mixture was slowly quenched with 10% aq. citric acid (200 mL) and extracted with EtOAc (3×250 mL). The combined organic layers were washed with brine (250 mL), dried over Na2SO4, evaporated and purified by column chromatography (PE/EtOAc=3/1) to give 6-chloro-4-formyl-N,N-diisopropylnicotinamide (24.0 g, 60% yield) as a yellow solid.
m/z ES+ [M+H]+ 269.0; 1H NMR (400 MHz, DMSO-d6) δ ppm 9.99 (s, 1H), 8.54 (s, 1H), 8.04 (s, 1H), 3.62-3.56 (m, 1H), 3.51-3.46 (m, 1H), 1.47 (d, J=6.4 Hz, 6H), 1.10 (d, J=6.4 Hz, 6H).
Step c. To a solution of 6-chloro-4-formyl-N,N-diisopropylnicotinamide (24.0 g, 89.3 mmol) in EtOH (500 mL) was slowly added NaBH4 (23.9 g, 634 mmol) at 25° C. The reaction mixture was stirred at 25° C. for 2 h. Upon completion, mixture was slowly poured into water (1000 mL) at 0° C., quenched with 1 M HCl (100 mL) and extracted with EtOAc (3×250 mL). The combined organic layers were washed with brine (300 mL), dried over (Na2SO4), evaporated and purified by column chromatography (PE/EtOAc=3/1) to give 6-chloro-4-(hydroxymethyl)-N,N-diisopropylnicotinamide (10.0 g, 33% yield) as a light yellow solid.
m/z ES+ [M+H]+ 271.1; 1H NMR (400 MHz, CDCl3) δ ppm 8.18 (s, 1H), 7.45 (s, 1H), 4.76-4.41 (m, 2H), 3.77-3.68 (m, 1H), 3.57-3.52 (m, 2H), 1.55 (d, J=6.4 Hz, 6H), 1.17 (s, 6H).
Step d. A solution of 6-chloro-4-(hydroxymethyl)-N,N-diisopropylnicotinamide (9.00 g, 33.2 mmol) in 6 M HCl (180 mL) was stirred at 100° C. for 1 h. Upon completion, the mixture was basified to pH 7-8 with sat. aq. Na2CO3 at 0° C. and extracted with EtOAc (3×100 mL). The combined organic layers were washed with brine (100 mL), dried over Na2SO4 and evaporated to give 6-chlorofuro[3,4-c]pyridin-3(1H)-one (3.00 g, 53% yield) as a yellow solid.
m/z ES+ [M+H]+ 170.0.
Step e. To a solution of 6-chlorofuro[3,4-c]pyridin-3(1H)-one (2.80 g, 16.5 mmol) in EtOH (60 mL) was slowly added NaBH4 (4.37 g, 115 mmol) in batches at 25° C. The reaction mixture was stirred at 25° C. for 2 h. Upon completion, the mixture was poured into water (100 mL), quenched with 1 M HCl (50 ml) and extracted with EtOAc (3×100 mL). The combined organic layers were dried over Na2SO4 and evaporated to give (6-chloropyridine-3,4-diyl)dimethanol (1.40 g, 48% yield) as a yellow solid.
m/z ES+ [M+H]+ 174.1.
Step f. To a solution of (6-chloropyridine-3,4-diyl)dimethanol (1.10 g, 6.34 mmol) in DCM (60 mL) was added SOCl2 (3.77 g, 31.6 mmol). The mixture was stirred at 50° C. for 2 h. Upon completion, the mixture was evaporated to give 2-chloro-4,5-bis(chloromethyl)pyridine (1.33 g, crude) as a yellow oil that was used directly without further purification.
Step g. To a solution of 2-chloro-4,5-bis(chloromethyl)pyridine (1.33 g, 6.32 mmol) in DCM (30 mL) was added DIPEA (2.45 g, 18.9 mmol) and (2,4-dimethoxyphenyl)methanamine (1.58 g, 9.48 mmol). The reaction mixture was stirred at 25° C. for 6 h. Upon completion, the mixture was evaporated and purified by column chromatography (PE/EtOAc=3/1) to give 6-chloro-2-(2,4-dimethoxybenzyl)-2,3-dihydro-1H-pyrrolo[3,4-c]pyridine (1.20 g, 57% yield) as a yellow solid.
m/z ES+ [M+H]+ 305.1; 1H NMR (400 MHz, CDCl3) δ ppm 8.18 (s, 1H), 7.26-7.23 (m, 1H), 7.15 (s, 1H), 6.51-6.48 (m, 2H), 3.95-3.93 (m, 4H), 3.87 (s, 2H), 3.83 (s, 6H).
Step h. A solution of 6-chloro-2-(2,4-dimethoxybenzyl)-2,3-dihydro-1H-pyrrolo[3,4-c]pyridine (1.00 g, 3.28 mmol) in TFA (3 mL) was stirred at 80° C. for 2 h. Upon completion, the mixture was evaporated to give 6-chloro-2,3-dihydro-1H-pyrrolo[3,4-c]pyridine (500 mg, 98% yield) as yellow oil that was used directly without further purification.
Step i. To a solution of 6-chloro-2,3-dihydro-1H-pyrrolo[3,4-c]pyridine (500 mg, 3.23 mmol) in DCM (10 mL) was added Boc2O (1.06 g, 4.85 mmol) and DIPEA (835 mg, 6.47 mmol). The reaction mixture was stirred at 25° C. for 6 h. Upon completion, the mixture was evaporated and purified by column chromatography (PE/EtOAc=3/1) to give tert-butyl 6-chloro-1,3-dihydro-2H-pyrrolo[3,4-c]pyridine-2-carboxylate (700 mg, 84% yield) as yellow solid
m/z ES+ [M+H]+ 255.0; 1H NMR (400 MHz, CDCl3) δ ppm 8.35-8.30 (m, 1H), 7.32-7.24 (m, 1H), 4.73-4.66 (m, 4H), 1.54 (s, 9H).
Steps j-k. These 2 steps were conducted in a similar manner to Intermediate B49, steps g-h.
m/z ES+ [M+H]+ 249.1.
Step a. To a solution of 3-bromo-6,7-dihydro-5H-pyrrolo[3,4-b]pyridine hydrochloride (CAS: 1394117-24-8; 1 g, 4.25 mmol) in DCM (15 mL) was added DIPEA (1.21 g, 9.34 mmol) and Boc2O (1.11 g, 5.10 mmol). The reaction mixture was stirred at 30° C. for 1 h. Upon completion, the mixture was evaporated and purified by column chromatography (PE/EtOAc=2/1) to give tert-butyl 3-bromo-5,7-dihydro-6H-pyrrolo[3,4-b]pyridine-6-carboxylate (1.2 g, 94% yield) as a white solid.
1H NMR (400 MHz, CDCl3) δ ppm 8.55 (s, 1H), 7.82-7.63 (m, 1H), 4.71-4.61 (m, 4H), 1.53 (s, 9H).
Step b. A mixture of tert-butyl 3-bromo-5,7-dihydro-6H-pyrrolo[3,4-b]pyridine-6-carboxylate (1 g, 3.34 mmol), Pd(OAc)2 (150 mg, 0.67 mmol), DPPP (276 mg, 0.67 mmol) and TEA (3.38 g, 33.4 mmol) in MeOH (10 mL) and DMSO (10 mL) was degassed and purged with N2 and CO 3 times. The reaction mixture was stirred at 60° C. for 12 h under a CO atmosphere (50 psi). Upon completion, the reaction mixture was evaporated, dissolved in EtOAc (100 mL) and washed with water (2×100 mL). The organic layer was dried over Na2SO4, evaporated and purified by column chromatography (PE/EtOAc=4/1) to give 6-(tert-butyl) 3-methyl 5,7-dihydro-6H-pyrrolo[3,4-b]pyridine-3,6-dicarboxylate (900 mg, 97% yield) as a white solid.
m/z ES+ [M+H]+ 279.1; 1H NMR (400 MHz, CDCl3) δ ppm 9.11 (s, 1H), 8.20-8.15 (m, 1H), 4.81-4.68 (m, 4H), 3.96 (s, 3H), 1.53 (s, 9H).
Step c. To a solution of 6-(tert-butyl) 3-methyl 5,7-dihydro-6H-pyrrolo[3,4-b]pyridine-3,6-dicarboxylate (0.1 g, 0.36 mmol) in DCM (4 mL) was added DIBAL-H (1 M in toluene, 1.08 mL) at −78° C. The reaction mixture was stirred at −78° C. for 30 min. Upon completion, the mixture was quenched with MeOH (0.5 mL) at −78° C. and the mixture was warmed to 20° C. until a solid was formed. The solid was filtered and the filtrate was evaporated and purified by column chromatography (PE/EtOAc=2/1) to give the title compound (0.1 g, crude) as a yellow oil.
1H NMR (400 MHz, CDCl3) δ ppm 10.12 (s, 1H), 8.96 (s, 1H), 8.08-8.03 (m, 1H), 4.82-4.75 (m, 4H), 1.54 (s, 9H).
Steps a-b. These 2 steps were conducted in a similar manner to Intermediate B49, steps g-h, using tert-butyl 2-chloro-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidine-6-carboxylate.
1H NMR (400 MHz, CDCl3) δ ppm 10.05 (s, 1H), 8.83-8.76 (m, 1H), 4.78-4.69 (m, 4H), 1.48 (s, 9H).
Step a. To a solution of 2-bromo-5-chlorobenzonitrile (CAS: 57381-37-0; 5 g, 23.1 mmol) in THF (100 mL) was added a solution of borane dimethyl sulfide complex (10 M in THF, 6.93 mL) at 0° C. The reaction mixture was stirred at 20° C. for 12 h. Upon completion, the mixture was quenched with MeOH (50 mL) at 0° C., evaporated and purified by column chromatography (PE/EtOAc=10/1) to give (2-bromo-5-chlorophenyl)methanamine (3 g, crude) as a yellow solid.
Step b. To a solution of (2-bromo-5-chlorophenyl)methanamine (8 g, 36.3 mmol) and Boc2O (9.50 g, 43.5 mmol) in DCM (200 mL) was added TEA (7.34 g, 72.5 mmol). The reaction mixture was stirred at 15° C. for 12 h. Upon completion, the mixture was quenched with water (40 mL) and extracted with EtOAc (2×30 mL). The combined organic layers were washed with brine (30 mL), dried over Na2SO4, evaporated and purified by column chromatography (PE/EtOAc=5/1) to give tert-butyl (2-bromo-5-chlorobenzyl)carbamate (8 g, 58% yield) as a white solid.
m/z ES+ [M-tBu+H]+ 265.9; 1H NMR (400 MHz, DMSO-d6) δ ppm 7.63 (d, J=8.4 Hz, 1H), 7.53 (t, J=5.6 Hz, 1H), 7.35-7.22 (m, 2H), 4.14 (d, J=6.0 Hz, 2H), 1.41 (s, 9H).
Step c. To a solution of tert-butyl (2-bromo-5-chlorobenzyl)carbamate (2 g, 6.24 mmol), acrylonitrile (1.66 g, 31.2 mmol) and t-Bu3P—Pd-G2 (CAS: 1375325-71-5; 160 mg, 0.31 mmol) in DMF (20 mL) was added N-cyclohexyl-N-methylcyclohexanamine (CAS: 7560-83-0; 1.34 g, 6.86 mmol). The reaction mixture was purged with N2 3 times and stirred at 110° C. for 16 h under a N2 atmosphere. Upon completion, the mixture was quenched with water (50 mL) and extracted with EtOAc (3×50 mL). The combined organic layers were washed with brine (60 mL), dried over Na2SO4, evaporated and purified by column chromatography (PE/EtOAc=3/1) to give tert-butyl (E)-(5-chloro-2-(2-cyanovinyl)benzyl)carbamate (1.4 g, 77% yield) as a white solid.
m/z ES+ [M+Na]+ 315.1; 1H NMR 400 MHz, CDCl3) δ ppm 7.79 (d, J=8.4 Hz, 1H), 7.61 (d, J=16.4 Hz, 1H), 7.35 (d, J=8.4 Hz, 1H), 5.74 (d, J=16.4 Hz, 1H), 5.50 (d, J=12.0 Hz, 1H), 4.75 (br s, 1H), 4.31-4.26 (m, 2H), 1.40 (s, 9H).
Step d. To a solution of tert-butyl (E)-(5-chloro-2-(2-cyanovinyl)benzyl)carbamate (300 mg, 1.02 mmol) in THF (3 mL) was added NaH (49 mg, 1.23 mmol, 60% dispersion in mineral oil) at 0° C. and the reaction mixture was stirred for at 0° C. for 30 min. Upon completion, the mixture was quenched with sat. aq. NH4Cl (0.1 mL), diluted with water (30 mL) and extracted with EtOAc (2×30 mL). The combined organic layers were washed with brine (30 mL), dried over Na2SO4, evaporated and purified by column chromatography (PE/EtOAc=3/1) to give tert-butyl 5-chloro-1-(cyanomethyl)isoindoline-2-carboxylate (180 mg, 60% yield) as a white solid.
1H NMR (400 MHz, CDCl3) δ ppm 7.37-7.27 (m, 3H), 5.23-5.15 (m, 1H), 4.80-4.63 (m, 2H), 3.29-3.22 (m, 1H), 3.00-2.90 (m, 1H), 1.53-1.51 (m, 9H).
Steps e-f. These 2 steps were conducted in a similar manner to Intermediate B49, steps g-h.
1H NMR (400 MHz, CDCl3) δ ppm 10.05 (s, 1H), 7.96-7.79 (m, 2H), 7.56 (d, J=8.0 Hz, 1H), 5.35-5.26 (m, 1H), 4.93-4.73 (m, 2H), 3.36-3.28 (m, 1H), 3.03-2.96 (m, 1H), 1.57-1.53 (m, 9H).
The title compound was prepared in a similar manner to Intermediate B57, using 2-bromo-4-chlorobenzonitrile (CAS: 57381-49-4) in step a.
1H NMR (400 MHz, CDCl3) δ ppm 10.05 (s, 1H), 7.91 (d, J=8.0 Hz, 1H), 7.88 (s, 1H), 7.56-7.44 (m, 1H), 5.36-5.23 (m, 1H), 4.93-4.73 (m, 2H), 3.46-3.06 (m, 1H), 3.00-2.90 (m, 1H), 1.57-1.51 (m, 9H).
Step a. A mixture of (3-bromo-6-chloropyridin-2-yl)methanol (CAS: 1227601-71-9; 1 g, 4.50 mmol) and MnO2 (3.91 g, 45.0 mmol) in DCM (5 mL) was stirred at 25° C. for 12 h under a N2 atmosphere. Upon completion, the mixture was evaporated and purified by column chromatography (PE/EtOAc=5/1) to give 3-bromo-6-chloropicolinaldehyde (300 mg, 30% yield) as a white solid.
1H NMR (400 MHz, CDCl3) δ ppm 10.04 (s, 1H), 7.92 (d, J=8.4 Hz, 1H), 7.33 (d, J=8.4 Hz, 1H).
Step b. A mixture of 3-bromo-6-chloropicolinaldehyde (800 mg, 3.63 mmol) and 2-(triphenyl-λ5-phosphanylidene)acetonitrile (CAS: 16640-68-9; 1.64 g, 5.44 mmol) in DCM (5 mL) was stirred at 25° C. for 4 h under a N2 atmosphere. Upon completion, the mixture was evaporated and purified by column chromatography (PE/EtOAc=5/1) to give (E)-3-(3-bromo-6-chloropyridin-2-yl)acrylonitrile (0.85 g, 96% yield) as a white solid.
1H NMR (400 MHz, CDCl3) δ ppm 7.91-7.87 (m, 1H), 7.79 (d, J=15.6 Hz, 1H), 7.24 (d, J=8.4 Hz, 1H), 6.72 (d, J=11.6 Hz, 1H).
Step c. A mixture of potassium (((tert-butoxycarbonyl)amino)methyl)trifluoroborate (CAS: 1314538-55-0; 535 mg, 2.26 mmol), (E)-3-(3-bromo-6-chloropyridin-2-yl)acrylonitrile (500 mg, 2.05 mmol), cataCXium-A-Pd-G3 (CAS: 1651823-59-4; 149 mg, 0.21 mmol) and Cs2CO3 (2.01 g, 6.16 mmol) in 1,4-dioxane (2 mL) and water (0.4 mL) was degassed and purged with N2 3 times. The reaction mixture was stirred at 100° C. for 7 h under a N2 atmosphere. Upon completion, the mixture was filtered, evaporated and purified by column chromatography (PE/EtOAc=3/1) to give tert-butyl (E)-((6-chloro-2-(2-cyanovinyl)pyridin-3-yl)methyl)carbamate (0.38 g, 63% yield) as a white solid.
m/z ES+ [M+H]+ 294.1; 1H NMR (400 MHz, CDCl3) δ ppm 7.69-7.63 (m, 2H), 6.96-6.72 (m, 2H), 4.75 (br s, 1H), 4.42 (d, J=6.0 Hz, 2H), 1.48 (s, 9H).
Steps d-f. These 3 steps were conducted in a similar manner to Intermediate B57, steps d-f.
1H NMR (400 MHz, CDCl3) δ ppm 10.06-9.92 (m, 1H), 7.91 (d, J=7.6 Hz, 1H), 7.83-7.69 (m, 1H), 5.21-5.10 (m, 1H), 4.88-4.70 (m, 2H), 3.61-3.21 (m, 1H), 3.08-3.06 (m, 1H), 1.51-1.45 (m, 9H).
Steps a-e. These 5 steps were conducted in a similar manner to Intermediate B59, steps b-f, using 2,6-dichloronicotinaldehyde (CAS: 55304-73-9) in step a.
1H NMR (400 MHz, CDCl3) δ ppm 10.22-9.97 (m, 1H), 8.00-7.92 (m, 2H), 5.39-5.28 (m, 1H), 4.82 (s, 2H), 3.30-3.03 (m, 2H), 1.58-1.55 (m, 9H).
Step a. A solution of isopropylmagnesium chloride lithium complex (1.3 M in THF, 31.2 mL) was added dropwise to a solution of 2,3-dibromo-5-chloropyridine (CAS: 137628-17-2; 10 g, 36.8 mmol) in THF (100 mL) at −40° C. under a N2 atmosphere. The mixture was stirred at −40° C. for 1 h, after which DMF (10 mL) was added dropwise and the reaction mixture was allowed to warm to 25° C. over 30 min. Upon completion, the mixture was quenched with 1 M HCl and extracted with EtOAc (2×80 mL). The combined organic layers were washed with brine (60 mL), dried over Na2SO4, evaporated and purified by column chromatography (PE/EtOAc=9/1) to give 2-bromo-5-chloronicotinaldehyde (9.5 g, 58% yield) as a pink solid.
1H NMR (400 MHz, CDCl3) δ ppm 10.29 (s, 1H), 8.55 (d, J=2.8 Hz, 1H), 8.13 (d, J=2.8 Hz, 1H).
Steps b-f. These 5 steps were conducted in a similar manner to Intermediate B59, steps b-f.
m/z ES+ [M+H]+ 288.1; 1H NMR (400 MHz, CDCl3) δ ppm 10.16 (s, 1H), 9.07 (s, 1H), 8.20 (s, 1H), 5.37-5.32 (m, 1H), 4.99-4.79 (m, 2H), 3.41-3.01 (m, 2H), 1.58-1.53 (m, 9H).
Intermediate B62: tert-butyl 7-(cyanomethyl)-3-formyl-5,7-dihydro-6H-pyrrolo[3,4-b]pyridine-6-carboxylate
Step a. To a solution of 3-bromo-5-chloropicolinic acid (CAS: 1189513-50-5; 10 g, 42.3 mmol) in MeOH (100 mL) was added H2SO4 (20.7 g, 211 mmol) at 0° C. The reaction mixture was stirred at 90° C. for 4 h. Upon completion, the mixture was quenched with water (500 mL) at 0° C. and extracted with EtOAc (2×500 mL). The combined organic layers were washed with brine (2×1 L), dried over Na2SO4 and evaporated to give the methyl 3-bromo-5-chloropicolinate (9.4 g, 89% yield) as a white solid.
1H NMR (400 MHz, DMSO-d6) δ ppm 8.72 (s, 1H), 8.56 (s, 1H), 3.91 (s, 3H).
Steps c-h. These 6 steps were conducted in a similar manner to Intermediate B59, steps a-f.
m/z ES+ [M+H]+ 288.2; 1H NMR (400 MHz, DMSO-d6) δ ppm 10.13 (s, 1H), 9.08 (s, 1H), 8.30 (d, J=6.0 Hz, 1H), 5.27-5.16 (m, 1H), 4.83-4.57 (m, 2H), 3.50-3.32 (m, 1H), 3.25-3.13 (m, 1H), 1.50 (s, 9H).
Step a. To a mixture of tert-butyl 3-amino-4-hydroxypyrrolidine-1-carboxylate (CAS: 190141-99-2; 37 g, 183 mmol) and NaHCO3 (46.1 g, 548.8 mmol) in THF (300 mL) and water (100 mL) was added CbzCl (37.5 g, 219 mmol). The reaction mixture was stirred at 25° C. for 12 h under a N2 atmosphere. Upon completion, the mixture was diluted with water (600 mL) and extracted with EtOAc (3×200 mL). The combined organic layers were washed with brine (200 mL), dried over Na2SO4, evaporated and purified by column chromatography (PE/EtOAc=5/1 to 1/1) to give tert-butyl 3-(((benzyloxy)carbonyl)amino)-4-hydroxypyrrolidine-1-carboxylate (60 g, 97% yield) as a yellow oil.
1H NMR (400 MHz, CDCl3) δ ppm 7.38-7.32 (m, 5H), 5.10 (s, 2H), 5.06-4.87 (m, 1H), 4.25 (br s, 1H), 4.05-3.92 (m, 1H), 3.85-3.75 (m, 1H), 3.68-3.55 (m, 1H), 3.50-2.98 (m, 3H), 1.45 (s, 9H).
Step b. To a solution of tert-butyl 3-(((benzyloxy)carbonyl)amino)-4-hydroxypyrrolidine-1-carboxylate (40 g, 118 mmol) and TEA (36.1 g, 356 mmol) in DCM (400 mL) was added MsCl (27.8 g, 242 mmol). The reaction mixture was stirred at 0° C. for 1 h. Upon completion, the mixture was quenched with water (500 mL) at 0° C. and extracted with DCM (2×300 mL). The combined organic layers were washed with brine (250 mL), dried over Na2SO4 and evaporated to give tert-butyl 3-(((benzyloxy)carbonyl)amino)-4-((methylsulfonyl)oxy)pyrrolidine-1-carboxylate (50 g, crude) as a yellow oil.
1H NMR (400 MHz, CDCl3) δ ppm 7.38-7.32 (m, 5H), 5.21-5.01 (m, 4H), 4.30-4.13 (m, 1H), 3.84-3.56 (m, 3H), 3.48-3.30 (m, 1H), 3.14 (s, 3H), 1.45 (s, 9H).
Step c. A mixture of tert-butyl 3-(((benzyloxy)carbonyl)amino)-4-((methylsulfonyl)oxy)pyrrolidine-1-carboxylate (25 g, 60.3 mmol) and NaN3 (19.3 g, 296.6 mmol) in DMF (250 mL) was stirred at 100° C. for 12 h under a N2 atmosphere. Upon completion, the mixture was quenched with sat. aq. NaHCO3 (50 mL) at 25° C., diluted with water (1000 mL) and extracted with EtOAc (3×200 mL). The combined organic layers were washed with brine (200 mL), dried over Na2SO4 and filtered to give a solution of tert-butyl 3-azido-4-(((benzyloxy)carbonyl)amino)pyrrolidine-1-carboxylate (21 g, crude) in EtOAc as a yellow liquid that was used directly in the next step without further purification.
m/z ES+ [M+Na]+ 384.0.
Step d. To a solution of tert-butyl 3-azido-4-(((benzyloxy)carbonyl)amino)pyrrolidine-1-carboxylate (21 g, 58.1 mmol) in EtOAc was added MeOH (500 mL) and 10% Pd/C (2 g) under a N2 atmosphere. The suspension was degassed, purged with H2 3 times and then stirred at 25° C. for 12 h under a H2 atmosphere (15 psi). Upon completion, the mixture was filtered and evaporated to give tert-butyl 3-amino-4-(((benzyloxy)carbonyl)amino)pyrrolidine-1-carboxylate (20 g, crude) as a yellow oil.
m/z ES+ [M+H]+ 336.1; 1H NMR (400 MHz, CDCl3) δ ppm 7.43-7.32 (m, 5H), 5.57-5.38 (m, 1H), 5.13 (s, 2H), 4.22-4.10 (m, 1H), 3.71-3.67 (m, 1H), 3.64-3.50 (m, 2H), 3.33-3.07 (m, 2H), 1.47 (s, 9H).
Step e. To a solution of tert-butyl 3-amino-4-(((benzyloxy)carbonyl)amino)pyrrolidine-1-carboxylate (40 g, 119 mmol) in MeOH (500 mL) was added 10% Pd/C (6 g) under a N2 atmosphere. The suspension was degassed and purged with H2 3 times and the reaction mixture was stirred at 25° C. for 12 h under a H2 atmosphere (40 psi). Upon completion, the reaction mixture was filtered and evaporated to give tert-butyl 3,4-diaminopyrrolidine-1-carboxylate (16 g, crude) as a yellow oil.
1H NMR (400 MHz, CDCl3) δ ppm 3.71-3.67 (m, 1H), 3.64-3.48 (m, 2H), 3.37-3.34 (m, 1H), 3.20-3.07 (m, 2H), 1.47 (s, 9H).
Step f. A mixture of tert-butyl 3,4-diaminopyrrolidine-1-carboxylate (5 g, 24.8 mmol) and ethyl 2-ethoxy-2-iminoacetate (CAS: 816-27-3; 3.61 g, 24.8 mmol) in HFIP (50 mL) was stirred at 60° C. for 12 h under a N2 atmosphere. Upon completion, the reaction mixture was evaporated to give 5-(tert-butyl) 2-ethyl 3a,4,6,6a-tetrahydropyrrolo[3,4-d]imidazole-2,5(1H)-dicarboxylate (7.04 g, crude) as a colourless oil.
m/z ES+ [M+H]+ 284.1.
To a solution of oxalyl chloride (7.42 mL, 84.7 mmol) in DCM (100 mL) at −65° C. was added DMSO (13.2 mL, 169 mmol). The mixture was stirred for 30 min, after which, the crude 5-(tert-butyl) 2-ethyl 3a,4,6,6a-tetrahydropyrrolo[3,4-d]imidazole-2,5(1H)-dicarboxylate (8 g, 28.2 mmol) in DCM (20 mL) was added at −65° C. The mixture was stirred for a further 30 min and then TEA (25.7 g, 254.1 mmol) was added and the reaction mixture was allowed to warm to 25° C. over 1 h. Upon completion, the reaction mixture was evaporated, diluted with water (100 mL) and extracted with EtOAc (3×50 mL). The combined organic layers were washed with brine (50 mL), dried over Na2SO4 and evaporated to give 5-(tert-butyl) 2-ethyl 4,6-dihydropyrrolo[3,4-d]imidazole-2,5(1H)-dicarboxylate (8 g, crude) as a yellow oil.
m/z ES+ [M+H]+ 282.1; 1H NMR (400 MHz, CDCl3) δ ppm 4.53-4.33 (m, 6H), 1.51-1.40 (m, 9H), 1.23-1.17 (m, 3H).
Step g. To a mixture of 5-(tert-butyl) 2-ethyl 4,6-dihydropyrrolo[3,4-d]imidazole-2,5(1H)-dicarboxylate (7 g, 24.8 mmol) and K2CO3 (10.3 g, 74.7 mmol) in DMF (100 mL) was added MeI (3.53 g, 24.9 mmol). The reaction mixture was stirred at 25° C. for 12 h under a N2 atmosphere. Upon completion, the mixture was quenched with water (200 mL) at 25° C. and extracted with EtOAc (3×100 mL). The combined organic layers were washed with brine (50 mL), dried over Na2SO4, evaporated and purified by column chromatography (PE/EtOAc=1/1 to 0/1) to give 5-(tert-butyl) 2-ethyl 1-methyl-4,6-dihydropyrrolo[3,4-d]imidazole-2,5(1H)-dicarboxylate (2.4 g, 33% yield) as a yellow solid.
m/z ES+ [M+H]+ 296.1; 1H NMR (400 MHz, CDCl3) δ ppm 4.52-4.38 (m, 6H), 3.96 (s, 3H), 1.51 (s, 9H), 1.46-1.42 (m, 3H).
Step h. To a solution of 5-(tert-butyl) 2-ethyl 1-methyl-4,6-dihydropyrrolo[3,4-d]imidazole-2,5(1H)-dicarboxylate (2.1 g, 7.11 mmol) in DCM (100 mL) was added DIBAL-H (1 M in toluene, 21.3 mL). The reaction mixture was stirred at −78° C. for 1 h. Upon completion, the mixture was quenched by with MeOH (4 mL) at −65° C., and then stirred at 25° C. for 1 h. The mixture was filtered and the filtrate was evaporated to give the title compound (1.1 g, 62% yield) as a yellow solid.
1H NMR (400 MHz, CDCl3) δ ppm 9.72 (s, 1H), 4.59-4.41 (m, 4H), 3.98 (d, J=2.8 Hz, 3H), 1.53 (s, 9H).
The title compound was prepared in a similar manner to Intermediate B63, using iodoethane in step g.
1H NMR (400 MHz, CDCl3) δ ppm 9.73 (s, 1H), 4.63-4.37 (m, 6H), 1.54 (s, 9H), 1.46-1.42 (m, 3H).
Step a. To a mixture of tert-butyl 2,5-dihydro-1H-pyrrole-1-carboxylate (5 g, 29.6 mmol) in water (2.5 mL) and DMSO (50 mL) was added NBS (6.84 g, 38.4 mmol) at 0° C. The reaction mixture was stirred at 25° C. for 2 h. Upon completion, the mixture was diluted with water (200 mL) and extracted with EtOAc (2×100 mL). The combined organic layers were washed with brine (200 mL), dried over Na2SO4, evaporated and purified by column chromatography (PE/EtOAc=1/1) to give tert-butyl (rel-trans)-3-bromo-4-hydroxypyrrolidine-1-carboxylate (6.85 g, 87% yield) as a colourless oil.
1H NMR (400 MHz, CDCl3) δ ppm 4.47 (s, 1H), 4.22-4.09 (m, 1H), 4.05-3.95 (m, 1H), 3.92-3.67 (m, 2H), 3.48-3.33 (m, 1H), 3.01-2.78 (m, 1H), 1.47 (s, 9H).
Step b. To a solution of tert-butyl (rel-trans)-3-bromo-4-hydroxypyrrolidine-1-carboxylate (6.85 g, 25.7 mmol) in DCM (70 mL) was added DMP (21.8 g, 51.5 mmol). The reaction mixture was stirred at 20° C. for 12 h. Upon completion, the mixture was filtered, evaporated and purified by column chromatography (PE/EtOAc=1/1) to give tert-butyl 3-bromo-4-oxopyrrolidine-1-carboxylate (6.4 g, 94% yield) as a black solid.
1H NMR (400 MHz, CDCl3) δ ppm 4.41-4.36 (m, 1H), 4.20-4.12 (m, 1H), 4.11-4.03 (m, 1H), 4.02-3.90 (m, 1H), 3.86-3.72 (m, 1H), 1.49 (s, 9H).
Step c. A mixture of tert-butyl 3-bromo-4-oxopyrrolidine-1-carboxylate (3 g, 11.4 mmol), ethyl 2-amino-2-thioxoacetate (CAS: 16982-21-1; 1.51 g, 11.4 mmol), PPTS (856 mg, 3.41 mmol) and 4 Å molecular sieves (6 g, 3.79 mmol) in toluene (50 mL) was stirred at 120° C. for 16 h under a N2 atmosphere. Upon completion, the reaction mixture was evaporated, diluted with water (30 mL) and extracted with EtOAc (30 mL). The organic layer was washed with brine (30 mL), dried over Na2SO4, evaporated and purified by column chromatography (PE/EtOAc=5/1) followed by reverse phase flash chromatography (water (0.1% FA)/MeCN) to give 5-(tert-butyl) 2-ethyl 4,6-dihydro-5H-pyrrolo[3,4-d]thiazole-2,5-dicarboxylate (740 mg, 22% yield) as a yellow solid.
m/z ES+ [M+H]+ 299.3; 1H NMR (400 MHz, CDCl3) δ ppm 4.79-4.72 (m, 2H), 4.70-4.65 (m, 1H), 4.63-4.58 (m, 1H), 4.53-4.45 (m, 2H), 1.53 (s, 9H), 1.49-1.43 (m, 3H).
Step d. This step was conducted in a similar manner to Intermediate B63, step h.
1H NMR (400 MHz, CDCl3) δ ppm 9.95 (s, 1H), 4.83-4.72 (m, 2H), 4.72-4.62 (m, 2H), 1.53 (s, 9H).
Step a. This step was conducted in a similar manner to Intermediate B1, step c, using tert-butyl 2-(hydroxymethyl)-4,6-dihydro-5H-pyrrolo[3,4-d]oxazole-5-carboxylate (CAS: 1251012-01-7) in step a.
1H NMR (400 MHz, CDCl3) δ ppm 9.71 (s, 1H), 4.61-4.57 (m, 2H), 4.51-4.46 (m, 2H), 1.53 (s, 9H).
Step a. To a solution of tert-butyl 4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxylate (CAS: 657428-42-7; 15.0 g, 71.7 mmol) in DCE (100 mL) was added NIS (24.2 g, 107.5 mmol). The reaction mixture was stirred at 80° C. for 12 h under a N2 atmosphere. Upon completion, the mixture was quenched with water (120 mL) and extracted with EtOAc (3×150 mL). The combined organic layers were washed with brine (150 mL), dried over Na2SO4, evaporated and purified by column chromatography (PE/EtOAc=1/1) to give tert-butyl 3-iodo-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxylate (1.50 g, 6% yield) as a yellow solid.
m/z ES+ [M+H]+ 336.0; 1H NMR (400 MHz, CDCl3) δ ppm 4.62-4.52 (m, 2H), 4.39-4.36 (m, 2H), 1.53-1.51 (m, 9H).
Step b. To a mixture of tert-butyl 3-iodo-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxylate (1.50 g, 4.48 mmol) and K2CO3 (1.86 g, 13.4 mmol) in DMF (6 mL) was added MeI (6.99 g, 49.2 mmol). The reaction mixture was stirred at 25° C. for 12 h under a N2 atmosphere. Upon completion, the mixture was quenched with water (60 mL) and extracted with EtOAc (2×100 mL). The combined organic layers were washed with brine (60 mL), dried over Na2SO4, and evaporated to give an inseparable mixture of tert-butyl 3-iodo-1-methyl-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxylate and tert-butyl 3-iodo-2-methyl-2,6-dihydropyrrolo[3,4-c]pyrazole-5(4H)-carboxylate (3:2 ratio; 1.2 g, 77% yield) as a red solid.
m/z ES+ [M+H]+ 349.9
Step c. A mixture of tert-butyl 3-iodo-1-methyl-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxylate and tert-butyl 3-iodo-2-methyl-2,6-dihydropyrrolo[3,4-c]pyrazole-5(4H)-carboxylate (1.15 g, 3.3 mmol), Pd(dppf)Cl2 (241 mg, 0.33 mmol) and TEA (3.33 g, 33 mmol) in MeOH (100 mL) was stirred at 50° C. for 16 h under a CO atmosphere (50 psi). Upon completion, the mixture was evaporated and purified by column chromatography (PE/EtOAc=1/1) to give two products. The regiochemistry was confirmed by 2D NMR.
Intermediate B67a: 5-(tert-butyl) 3-methyl 1-methyl-4,6-dihydropyrrolo[3,4-c]pyrazole-3,5(1H)-dicarboxylate (450 mg, 49% yield) as a red solid.
m/z ES+ [M+H]+ 282.2; 1H NMR (400 MHz, CDCl3) δ ppm 4.58-4.48 (m, 4H), 3.92 (s, 3H), 3.89 (s, 3H), 1.51 (s, 9H).
Intermediate B67b: 5-(tert-butyl) 3-methyl 2-methyl-2,6-dihydropyrrolo[3,4-c]pyrazole-3,5(4H)-dicarboxylate (270 mg, 29% yield) as a white solid.
m/z ES+ [M+H]+ 282.2; 1H NMR (400 MHz, CDCl3) δ ppm 4.58-4.43 (m, 4H), 4.18 (s, 3H), 3.88 (d, J=12.8 Hz, 3H), 1.52 (d, J=3.2 Hz, 9H).
Steps a. This step was conducted in a similar manner to Intermediate B53, step c, using Intermediate B67a.
m/z ES+ [M+H]+ 252.3; 1H NMR (400 MHz, CDCl3) δ ppm 9.89 (s, 1H), 4.62-4.47 (m, 4H), 3.92 (s, 3H), 1.50 (s, 9H).
Step a. To a solution of 5-bromopyrazin-2-amine (59489-71-3; 15 g, 86.2 mmol) in 1,4-dioxane (150 mL) was added NaHCO3 (21.7 g, 259 mmol) and ethyl 3-bromo-2-oxopropanoate (25.2 g, 129 mmol). The reaction mixture was stirred at 100° C. for 16 h. Upon completion, the mixture was filtered, evaporated and purified by column chromatography (EtOAc to EtOAc/MeOH=20/1) to give ethyl 6-bromoimidazo[1,2-a]pyrazine-2-carboxylate (11.5 g, 49% yield) was obtained as yellow solid.
m/z ES+ [M+H]+ 272.0; 1H NMR (400 MHz, CDCl3) δ ppm 9.01 (s, 1H), 8.31 (s, 1H), 8.24 (s, 1H), 4.53-4.47 (m, 2H), 1.46 (t, J=7.2 Hz, 3H).
Step b. A mixture of ethyl 6-bromoimidazo[1,2-a]pyrazine-2-carboxylate (10.5 g, 38.9 mmol), potassium trifluoro(vinyl)borate (CAS: 13682-77-4; 52.1 g, 389 mmol), Cs2CO3 (38.0 g, 117 mmol) and XPhos-Pd-G2 (3.06 g, 3.89 mmol) in 1,4-dioxane (150 mL) and water (150 mL) was degassed and purged with N2 3 times. The reaction mixture was stirred at 100° C. for 16 h under a N2 atmosphere. Upon completion, the mixture diluted with water (800 mL) and extracted with EtOAc (3×300 mL). The combined organic layers were dried over Na2SO4, evaporated and purified by column chromatography (PE/EtOAc=4/1 to 2/1) to give ethyl 6-vinylimidazo[1,2-a]pyrazine-2-carboxylate (5.5 g, 65% yield) as a yellow solid.
1H NMR (400 MHz, CDCl3) δ ppm 9.17 (s, 1H), 8.21 (s, 1H), 7.96 (s, 1H), 6.74-6.67 (m, 1H), 6.39-6.35 (m, 1H), 5.56-5.53 (m, 1H), 4.52-4.46 (m, 2H), 1.46 (t, J=7.2 Hz, 3H).
Step c. This step was conducted in a similar manner to Intermediate B49, step h.
m/z ES+ [M+H]+ 220.0; 1H NMR (400 MHz, CDCl3) δ ppm 10.17 (s, 1H), 9.29 (s, 1H), 8.81 (s, 1H), 8.41 (s, 1H), 4.55-4.49 (m, 2H), 1.48 (t, J=7.2 Hz, 3H).
Step d. To a solution of ethyl 6-formylimidazo[1,2-a]pyrazine-2-carboxylate (2 g, 9.12 mmol) in DCM (100 mL) was added DAST (5.88 g, 36.5 mmol) at −78° C. The reaction mixture was allowed to warm to 0° C. over 16 h. Upon completion, the mixture was diluted with sat. aq. NaHCO3 (50 mL) and extracted with EtOAc (3×20 mL). The combined organic layers were washed with brine (3×20 mL), dried over Na2SO4, evaporated and purified by column chromatography (PE/EtOAc=3/1 to 1/1) to give ethyl 6-(difluoromethyl)imidazo[1,2-a]pyrazine-2-carboxylate (1.24 g, 56% yield) as a yellow solid.
1H NMR (400 MHz, CDCl3) δ ppm 9.22 (s, 1H), 8.46 (s, 1H), 8.36 (s, 1H), 6.95-6.65 (m, 1H), 4.54-4.48 (m, 2H), 1.47 (t, J=7.2 Hz, 3H).
Step e. This step was conducted in a similar manner to Intermediate B63, step h.
1H NMR (400 MHz, CDCl3) δ ppm 9.76 (s, 1H), 7.47 (s, 1H), 6.26 (d, J=1.6 Hz, 1H), 6.21-5.92 (m, 1H), 4.89-4.84 (m, 1H), 4.62 (d, J=2.0 Hz, 2H).
Step f. To a solution of 6-(difluoromethyl)-5,6-dihydroimidazo[1,2-a]pyrazine-2-carbaldehyde (660 mg, 3.31 mmol) in DCM (50 mL) was added MnO2 (5.76 g, 66.3 mmol). The reaction mixture was stirred at 25° C. for 16 h. Upon completion, the mixture was filtered and evaporated to give the title compound (490 mg, crude) as a red solid.
1H NMR (400 MHz, CDCl3) δ ppm 10.24 (s, 1H), 9.25 (s, 1H), 8.50 (s, 1H), 8.32 (s, 1H), 6.96-6.65 (m, 1H).
Step a. A mixture of (5-chloropyrazin-2-yl)methanol (CAS: 72788-94-4; 8.4 g, 58.1 mmol), diphenylmethanimine (12.6 g, 69.7 mmol), Pd2(dba)3 (2.66 g, 2.91 mmol), Xantphos (3.36 g, 5.81 mmol) and Cs2CO3 (56.8 g, 174 mmol) in 1,4-dioxane (180 mL) was degassed and purged with N2 3 times. The reaction mixture was stirred at 110° C. for 36 h under a N2 atmosphere. Upon completion, the reaction mixture was diluted with water (200 mL) and extracted with EtOAc (3×100 mL). The combined organic layers were dried over Na2SO4, evaporated and purified by column chromatography (PE/EtOAc=4/1 to 1/1) to give (5-((diphenylmethylene)amino)pyrazin-2-yl)methanol (3.32 g, 20% yield) as a yellow oil.
m/z ES+ [M+H]+ 290.1; 1H NMR (400 MHz, CDCl3) δ ppm 8.22 (d, J=1.2 Hz, 1H), 7.83 (d, J=1.2 Hz, 1H), 7.75 (d, J=7.6 Hz, 2H), 7.51-7.43 (m, 1H), 7.37 (t, J=7.2 Hz, 2H), 7.31-7.20 (m, 3H), 7.09 (d, J=6.8 Hz, 2H), 4.65 (d, J=4.4 Hz, 2H), 2.75-2.65 (m, 1H).
Step b. To a solution of 5-((diphenylmethylene)amino)pyrazin-2-yl)methanol (2.1 g, 7.26 mmol) in DCM (30 mL) was added TFA (6 mL). The reaction mixture was stirred at 20° C. for 16 h. Upon completion, the mixture was evaporated to give (5-aminopyrazin-2-yl)methanol (1.8 g, crude) as a red oil.
1H NMR (400 MHz, CDCl3) δ ppm 8.50 (s, 1H), 7.95 (s, 1H), 5.39 (s, 2H).
Step c. This step was conducted in a similar manner to Intermediate B50a and Intermediate B50b, step a, using TBDPSCl.
m/z ES+ [M+H]+ 364.2; 1H NMR (400 MHz, CDCl3) δ ppm 8.20 (s, 1H), 7.89 (d, J=1.2 Hz, 1H), 7.75-7.65 (m, 4H), 7.46-7.41 (m, 2H), 7.41-7.35 (m, 4H), 4.77 (s, 2H), 4.54 (s, 2H), 1.10 (s, 9H).
Step d. This step was conducted in a similar manner to Intermediate B69, step a.
m/z ES+ [M+H]+ 460.3; 1H NMR (400 MHz, CDCl3) δ ppm 9.07 (s, 1H), 8.28 (s, 1H), 8.23 (d, J=1.6 Hz, 1H), 7.71-7.69 (m, 4H), 7.49-7.43 (m, 2H), 7.43-7.37 (m, 4H), 4.92 (d, J=1.6 Hz, 2H), 4.53-4.47 (m, 2H), 1.47 (t, J=7.2 Hz, 3H), 1.16 (s, 9H).
Step e. To a solution of ethyl 6-(((tert-butyldiphenylsilyl)oxy)methyl)imidazo[1,2-a]pyrazine-2-carboxylate (130 mg, 0.28 mmol) in THF (5 mL) was added LiBH4 (19 mg, 0.85 mmol) at 0° C. The reaction mixture was allowed to warm to 20° C. over 5 h. Upon completion, the reaction mixture was quenched with sat. aq. NH4Cl (0.5 mL) at 0° C., diluted with water (10 mL) and extracted with EtOAc (3×5 mL). The combined organic layers were dried over Na2SO4 and evaporated to give (6-(((tert-butyldiphenylsilyl)oxy)methyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-2-yl)methanol (120 mg, crude) as a light yellow solid.
m/z ES+ [M+H]+ 422.3.
Step f. To a solution of (6-(((tert-butyldiphenylsilyl)oxy)methyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-2-yl)methanol (120 mg, 0.29 mmol) in DCE (5 mL) was added MnO2 (990 mg, 11.4 mmol). The reaction mixture was stirred at 60° C. for 16 h. Upon completion, the reaction mixture was filtered and evaporated to give the title compound (80 mg, crude) as a light yellow solid.
m/z ES+ [M+H]+ 418.2; 1H NMR (400 MHz, CDCl3) δ ppm 9.91 (s, 1H), 8.38 (s, 1H), 7.64-7.16 (m, 4H), 7.45-7.38 (m, 7H), 4.40-4.31 (m, 1H), 4.21-4.10 (m, 3H), 3.74-3.70 (m, 1H), 1.06 (s, 9H).
Step a. This step was conducted in a similar manner to Intermediate B69, step a, using 5-(trifluoromethyl)pyrazin-2-amine (CAS: 69816-38-2).
m/z ES+ [M+H]+ 260.0; 1H NMR (400 MHz, CDCl3) δ ppm 8.97 (s, 1H), 8.78 (s, 1H), 8.39 (s, 1H), 4.26 (d, J=7.2 Hz, 2H), 1.26-1.19 (m, 3H).
Step b. To a solution of ethyl 6-(trifluoromethyl)imidazo[1,2-a]pyrazine-2-carboxylate (5.8 g, 22.4 mmol) and acetic acid (4.03 g, 67.1 mmol) in EtOH (100 mL) was added PtO2 (508 mg, 2.24 mmol) under a N2 atmosphere. The suspension was degassed and purged with H2 and the mixture was stirred at 50° C. for 12 h under a H2 atmosphere (50 psi). Upon completion, the mixture was filtered and evaporated to give ethyl 6-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazine-2-carboxylate (5.8 g, crude) as a yellow oil.
m/z ES+ [M+H]+ 264.1.
Steps c-d. These 2 steps were conducted in a similar manner to Intermediate B40, steps b-c.
m/z ES+ [M+H]+ 320.2; 1H NMR (400 MHz, CDCl3) δ ppm 9.94 (s, 1H), 7.68 (s, 1H), 5.50-5.15 (m, 2H), 4.61-4.42 (m, 3H), 1.61 (s, 9H).
Step a. To a solution of dimethyl 1H-pyrazole-3,5-dicarboxylate (5 g, 27.1 mmol), tert-butyl (1-hydroxypropan-2-yl)carbamate (5.71 g, 32.5 mmol) and PPh3 (14.2 g, 54.3 mmol) in THF (10 mL) under a N2 atmosphere was added DEAD (7.09 g, 40.7 mmol). The reaction mixture was stirred at 25° C. for 12 h. Upon completion, the mixture was evaporated and purified by column chromatography (PE/EtOAc=10/1 to 2/1) to give dimethyl 1-(2-((tert-butoxycarbonyl)amino)propyl)-1H-pyrazole-3,5-dicarboxylate (9.0 g, 97% yield) as a white solid.
1H NMR (400 MHz, CDCl3) δ ppm 7.35 (s, 1H), 4.81-4.65 (m, 2H), 4.58-4.45 (m, 1H), 4.27-4.21 (m, 1H), 3.93 (d, J=6.8 Hz, 6H), 1.30 (s, 9H), 1.20 (d, J=6.8 Hz, 3H).
Steps b-e. These 4 steps were conducted in a similar manner to Intermediate B33, steps b-e.
m/z ES+ [M+H]+ 296.2; 1H NMR (400 MHz, CDCl3) δ ppm 6.65 (s, 1H), 5.00-4.85 (m, 2H), 4.35-4.24 (m, 2H), 4.21-4.16 (m, 1H), 3.93 (s, 3H), 1.50 (s, 9H), 1.26 (t, J=7.2 Hz, 3H).
Step f. This step was conducted in a similar manner to Intermediate B3, step b.
m/z ES+ [M+H]+ 265.9; 1H NMR (400 MHz, CDCl3) δ ppm 9.95 (s, 1H), 6.64 (s, 1H), 5.03-4.88 (m, 2H), 4.35-4.30 (m, 2H), 4.20-4.16 (m, 1H), 1.51 (s, 9H), 1.20 (d, J=7.2 Hz, 3H).
The title compound was prepared in a similar manner to Intermediate B71, using 5-nitropyridin-2-amine (CAS: 4214-76-0) in step a.
1H NMR (400 MHz, CDCl3) δ ppm 9.82 (s, 1H), 7.49 (s, 1H), 4.75 (d, J=5.6 Hz, 1H), 4.33-4.29 (m, 1H), 4.20 (br s, 1H), 3.93-3.88 (m, 1H), 3.10-3.00 (m, 2H), 2.25-2.06 (m, 2H), 1.46 (s, 9H).
The title compound was prepared in a similar manner to Intermediate B71, using 4-nitropyridin-2-amine (CAS: 4487-50-7) in step a.
1H NMR (400 MHz, CDCl3) δ ppm 9.87 (s, 1H), 7.57 (s, 1H), 4.73 (s, 1H), 4.30-4.15 (m, 3H), 3.34-3.27 (m, 1H), 2.90-2.84 (m, 1H), 2.35-2.30 (m, 1H), 2.25-2.13 (m, 1H), 1.51 (s, 9H).
Step a. A solution of n-BuLi (2.5 M in THF, 1.15 mL) was added dropwise to a solution of tert-butyl 6-bromo-2,3-dihydro-4H-benzo[b][1,4]oxazine-4-carboxylate (CAS: 719310-31-3; 600 mg, 1.91 mmol) in THF (20 mL) at −70° C. under a N2 atmosphere. The reaction mixture was stirred at −70° C. for 30 min, after which DMF (1.47 mL, 19.1 mmol) in THF (10 mL) was added dropwise at −70° C. The resulting mixture was stirred at −70° C. for a further 1 h. Upon completion, the reaction mixture was quenched with water (10 mL) and extracted with EtOAc (5×2 mL). The combined organic layers were dried over Na2SO4, filtered, evaporated and purified by column chromatography (PE/EtOAc=7/1) to give the title compound (450 mg, 90% yield) as a light yellow oil.
1H NMR (400 MHz, CDCl3) δ ppm 9.87 (s, 1H), 8.38 (br s, 1H), 7.55 (d, J=8.4 Hz, 1H), 6.99 (d, J=8.4 Hz, 1H), 4.35-4.30 (m, 2H), 3.93-3.88 (m, 2H), 1.58 (s, 9H).
Steps a-c. These 3 steps were conducted in a similar manner to Intermediate B33, steps e-g, using methyl 3,4-dihydro-2H-benzo[b][1,4]oxazine-7-carboxylate (CAS: 142166-01-6) in step a.
1H NMR (400 MHz, CDCl3) δ ppm 9.79 (s, 1H), 7.99 (d, J=8.4 Hz, 1H), 7.38-7.27 (m, 2H), 4.22-4.18 (m, 2H), 3.85-3.82 (m, 2H), 1.49 (s, 9H).
Steps a-b. These 2 steps were conducted in a similar manner to Intermediate B40, steps b-c, using methyl 5-aminopicolinate (CAS: 67515-76-8) in step a.
1H NMR (400 MHz, CDCl3) δ ppm 9.92 (s, 1H), 8.53 (d, J=2.0 Hz, 1H), 8.12 (d, J=8.0 Hz, 1H), 7.89 (d, J=8.8 Hz, 1H), 6.77 (s, 1H), 1.48 (s, 9H).
Step a. To a mixture of 2-chloropyrimidin-5-amine (CAS: 56621-90-0; 0.5 g, 3.86 mmol) and NaHCO3 (973 mg, 11.6 mmol) in THF (5 mL) and water (2 mL) was added CbzCl (988 mg, 5.79 mmol). The reaction mixture was stirred at 25° C. for 12 h. Upon completion, the mixture was evaporated and the residue was purified by column chromatography (PE/EtOAc=2/1) to give benzyl (2-chloropyrimidin-5-yl)carbamate (0.7 g, 69% yield) as a white solid.
m/z ES+ [M+H]+ 264.0.
Steps b-c. These 2 steps were conducted in a similar manner to Intermediate B49, steps g-h.
m/z ES+ [M+H]+ 258.0; 1H NMR (400 MHz, CDCl3) δ ppm 10.05 (s, 1H), 9.10 (s, 2H), 7.46-7.39 (m, 5H), 7.12 (s, 1H), 5.28 (s, 2H).
Step a. This step was conducted in a similar manner to Intermediate B40, steps b, using 4-bromo-2,6-difluoroaniline (CAS: 67567-26-4).
1H NMR (400 MHz, CDCl3) δ ppm 7.19-7.11 (m, 2H), 5.85 (br s, 1H), 1.45 (s, 9H).
Steps b-c. These 2 steps were conducted in a similar manner to Intermediate B55, steps b-c.
1H NMR (400 MHz, CDCl3) δ ppm 9.90 (t, J=1.6 Hz, 1H), 7.53-7.43 (m, 2H), 6.19 (br s, 1H), 1.53 (s, 9H).
The title compound was prepared in a similar manner to Intermediate B27, using tert-butyl 3-fluoro-3-(hydroxymethyl)azetidine-1-carboxylate (CAS: 1126650-66-5) in step a.
1H NMR (400 MHz, CDCl3) δ ppm 9.79 (s, 1H), 4.31-4.28 (m, 1H), 4.18-4.14 (m, 1H), 4.13-4.03 (m, 1H), 3.97-3.90 (m, 2H), 3.65-3.52 (m, 3H), 1.54 (s, 9H).
Step a. A mixture of 5-bromo-1,3-dihydro-2H-inden-2-one (CAS: 174349-93-0; 0.5 g, 2.37 mmol), (2,4-dimethoxyphenyl)methanamine (594 mg, 3.55 mmol), NaBH(OAc)3 (1.00 g, 4.74 mmol) and acetic acid (14 mg, 0.24 mmol) in DCE (5 mL) was degassed and purged with N2 3 times. The reaction mixture was stirred at 25° C. for 12 h under a N2 atmosphere. Upon completion, the mixture was quenched with sat. aq. NH4Cl (1 mL), diluted with water (30 mL) and extracted with EtOAc (2×30 mL). The combined organic layers were washed with brine (30 mL), dried over Na2SO4 and evaporated to give 5-bromo-N-(2,4-dimethoxybenzyl)-2,3-dihydro-1H-inden-2-amine (140 mg, 16% yield) as a brown oil.
m/z ES+ [M+H]+ 362.0.
Step b. To a solution of 5-bromo-N-(2,4-dimethoxybenzyl)-2,3-dihydro-1H-inden-2-amine (4.3 g, 11.9 mmol), TEA (2.40 g, 23.7 mmol) and DMAP (145 mg, 1.19 mmol) in DCM (70 mL) was added Boc2O (3.89 g, 17.80 mmol). The reaction mixture was stirred at 20° C. for 1 h. Upon completion, the mixture was evaporated and purified by column chromatography (PE/EtOAc=5/1) to give tert-butyl (5-bromo-2,3-dihydro-1H-inden-2-yl)(2,4-dimethoxybenzyl)carbamate (1.7 g, 29% yield) as a brown oil.
m/z ES+ [M+Na]+ 484.0; 1H NMR (400 MHz, CDCl3) δ ppm 7.25-7.19 (m, 2H), 7.10 (d, J=8.0 Hz, 1H), 6.97 (d, J=8.0 Hz, 1H), 6.50-6.45 (m, 1H), 6.42 (d, J=2.4 Hz, 1H), 5.10-4.56 (m, 1H), 4.39 (s, 2H), 3.82 (s, 3H), 3.75 (s, 3H), 3.08-2.99 (m, 2H), 2.97 (d, J=8.4 Hz, 2H), 1.41 (s, 9H).
Step c. This step was conducted in a similar manner to to Intermediate B55, step b.
m/z ES+ [M+Na]+ 464.3; 1H NMR (400 MHz, CDCl3) δ ppm 7.79 (d, J=2.8 Hz, 1H), 7.25-7.20 (m, 1H), 7.16 (d, J=7.6 Hz, 1H), 7.12 (d, J=8.8 Hz, 1H), 6.50-6.45 (m, 1H), 6.42 (d, J=2.0 Hz, 1H), 4.48-4.33 (m, 2H), 3.89 (s, 3H), 3.82 (s, 3H), 3.77-3.73 (m, 3H), 3.14-2.91 (m, 5H), 1.42-1.37 (m, 9H).
Step d. A mixture of methyl 2-((tert-butoxycarbonyl)(2,4-dimethoxybenzyl)amino)-2,3-dihydro-1H-indene-5-carboxylate (900 mg, 2.04 mmol), 10% Pd(OH)2 (286 mg, 2.04 mmol) and 10% Pd/C (217 mg, 2.04 mmol) in MeOH (5 mL) was degassed and purged with H2 3 times. The reaction mixture was stirred at 25° C. for 12 h under a H2 atmosphere (15 psi). Upon completion, the mixture was filtered, evaporated and purified by column chromatography (PE/EtOAc=6/1) followed by prep-HPLC to give methyl 2-((tert-butoxycarbonyl)amino)-2,3-dihydro-1H-indene-5-carboxylate (190 mg, 32% yield) as a white solid.
m/z ES+ [M-tBu+H]+ 236.1; 1H NMR (400 MHz, CDCl3) δ ppm 8.00-7.78 (m, 2H), 7.30 (s, 1H), 4.81-4.68 (m, 1H), 4.60-4.46 (m, 1H), 3.92 (s, 3H), 3.42-3.25 (m, 2H), 2.98-2.73 (m, 2H), 1.47 (s, 9H).
Steps e-f. These 2 steps were conducted in a similar manner to Intermediate B5, steps b-c.
m/z ES+ [M-tBu+H]+ 206.0; 1H NMR (400 MHz, CDCl3) δ ppm 9.89 (s, 1H), 7.71-7.58 (m, 2H), 7.30 (d, J=7.6 Hz, 1H), 4.69-4.67 (m, 1H), 4.50-4.45 (m, 1H), 3.40-3.20 (m, 2H), 2.83-2.72 (m, 2H), 1.38 (s, 9H).
The title compound was prepared in a similar manner to Intermediate B35, using 4-aminobutan-1-ol (CAS: 13325-10-5) in step a.
1H NMR (400 MHz, CDCl3) δ ppm 9.71 (s, 1H), 7.80-7.76 (m, 2H), 7.67-7.64 (m, 2H), 3.70-3.66 (m, 2H), 2.47 (t, J=7.2 Hz, 2H), 2.00-1.97 (m, 2H).
Step a. To a solution of Intermediate B36 (770 mg, 3.33 mmol) and proline (115 mg, 1.00 mmol) in MeCN (10 mL) was added Selectfluor (1.77 g, 4.99 mmol) followed by TFA (114 mg, 1.00 mmol) at 0° C. The reaction mixture was stirred at 25° C. for 16 h. Upon completion, the mixture was diluted with sat. aq. NaHCO3 (20 mL) and extracted with EtOAc (3×20 mL). The combined layers were washed with brine (50 mL), dried over Na2SO4 and evaporated to give the title compound (800 mg, crude) as a yellow oil.
m/z ES+ [M+H]+ 250.3; 1H NMR (400 MHz, CH3OD) δ ppm 9.77 (d, J=5.6 Hz, 1H), 7.90-7.79 (m, 2H), 7.77-7.67 (m, 2H), 5.16-4.98 (m, 1H), 3.80-3.64 (m, 2H), 2.05-1.65 (m, 4H).
Step a. This step was conducted in a similar manner to Intermediate B35, step b, using 3-(benzyloxy)propan-1-ol (CAS: 4799-68-2).
1H NMR (400 MHz, CDCl3) δ ppm 9.73 (t, J=1.6 Hz, 1H), 7.32-7.27 (m, 2H), 7.27-7.19 (m, 3H), 4.47 (s, 2H), 3.75 (t, J=6.0 Hz, 2H), 2.66-2.60 (m, 2H).
Step a. This step was conducted in a similar manner to Intermediate B55, step a, using 3-fluoropiperidine hydrochloride (CAS: 737000-77-0).
1H NMR (400 MHz, CDCl3) δ ppm 4.73-4.46 (m, 1H), 3.70-3.39 (m, 3H), 3.38-3.19 (m, 1H), 1.95-1.75 (m, 3H), 1.52-1.43 (m, 10H).
Step b. To a mixture of RuCl3 (1.78 g, 8.56 mmol) and NaIO4 (30.5 g, 143 mmol) in water (160 mL) was added a solution of tert-butyl 3-fluoropiperidine-1-carboxylate (5.8 g, 28.5 mmol) in EtOAc (230 mL). The reaction mixture was stirred at 25° C. for 12 h. Upon completion, the reaction mixture was quenched with sat. aq. Na2S2O3 (200 mL), diluted with water (150 mL) and extracted with EtOAc (3×250 mL). The combined organic layers were dried over Na2SO4, evaporated and purified by column chromatography (PE/EtOAc=5/1) to give tert-butyl 5-fluoro-2-oxopiperidine-1-carboxylate (4.4 g, 71% yield) as a colourless oil.
1H NMR (400 MHz, CDCl3) δ ppm 5.17-4.96 (m, 1H), 4.19-4.05 (m, 1H), 3.78-3.59 (m, 1H), 2.79-2.65 (m, 1H), 2.56-2.49 (m, 1H), 2.25-2.12 (m, 1H), 2.02-1.87 (m, 1H), 1.54 (s, 9H).
Step c. To a solution of tert-butyl 5-fluoro-2-oxopiperidine-1-carboxylate (2.00 g, 9.21 mmol) in MeOH (20 mL) was added NaOMe (995 mg, 18.4 mmol). The reaction mixture was stirred at 25° C. for 30 min. Upon completion, the mixture was quenched with 1 M HCl (20 mL) and extracted with EtOAc (2×40 mL). The combined organic layers were washed with brine (40 mL), dried over Na2SO4, evaporated and purified by column chromatography (PE/EtOAc=5/1) to give methyl 5-((tert-butoxycarbonyl)amino)-4-fluoropentanoate (1.1 g, 48% yield) as a colourless oil.
1H NMR (400 MHz, CDCl3) δ ppm 4.86 (br s, 1H), 4.72-4.46 (m, 1H), 3.69 (s, 3H), 3.58-3.35 (m, 1H), 3.31-3.13 (m, 1H), 2.60-2.39 (m, 2H), 2.03-1.85 (m, 2H), 1.45 (s, 9H).
Step d. This step was conducted in a similar manner to Intermediate B3, step b.
1H NMR (400 MHz, CDCl3) δ ppm 9.81 (s, 1H), 4.86 (br s, 1H), 4.73-4.69 (m, 1H), 3.54-3.48 (m, 2H), 3.25-3.10 (m, 2H), 2.03-1.85 (m, 2H) 1.45 (s, 9H).
The title compound was prepared in a similar manner to Intermediate B88, using 3,3-difluoropiperidine hydrochloride (CAS: 496807-97-7) in step a.
1H NMR (400 MHz, CDCl3) δ ppm 9.83 (s, 1H), 4.85 (br s, 1H), 3.54 (d, J=6.8 Hz, 2H), 2.76 (t, J=7.6 Hz, 2H), 2.30-2.13 (m, 2H), 1.47 (s, 9H).
The title compound was prepared as described in Intermediate B30, step a.
1H NMR (400 MHz, CDCl3) δ ppm 9.78 (s, 1H), 4.13 (t, J=8.4 Hz, 2H), 3.61-3.52 (m, 2H), 2.99-2.87 (m, 1H), 2.86-2.80 (m, 2H), 1.44 (s, 9H).
The title compound was prepared in a similar manner to Intermediate B43, using tert-butyl 3-(3-hydroxypropyl)azetidine-1-carboxylate (CAS: 158602-43-8).
1H NMR (400 MHz, CDCl3) δ ppm 9.79 (t, J=1.2 Hz, 1H), 4.01 (t, J=8.4 Hz, 2H), 3.59-3.50 (m, 2H), 2.58-2.48 (m, 1H), 2.45-2.43 (m, 2H), 1.97-1.87 (m, 2H), 1.44 (s, 9H).
Step a. A mixture of ethyl 2-(diethoxyphosphoryl)-2-fluoroacetate (CAS: 2356-16-3; 10 g, 41.3 mmol), tert-butyl 3-oxoazetidine-1-carboxylate (CAS: 398489-26-4; 8.48 g, 49.6 mmol) and DBU (9.43 g, 61.9 mmol) in THF (100 mL) was degassed and purged with N2 3 times. The reaction mixture was stirred at 25° C. for 1 h under a N2 atmosphere. Upon completion, the mixture was evaporated and purified by column chromatography (PE/EtOAc=1/1) to give tert-butyl 3-(2-ethoxy-1-fluoro-2-oxoethylidene)azetidine-1-carboxylate (7.5 g, 70% yield) as a colourless oil.
1H NMR (400 MHz, CDCl3) δ ppm 4.71-4.63 (m, 2H), 4.58-4.56 (m, 2H), 4.22 (q, J=7.2 Hz, 2H), 1.39 (s, 9H), 1.26 (t, J=7.2 Hz, 3H).
Step b. This step was conducted in a similar manner to Intermediate B3, step b.
1H NMR (400 MHz, CDCl3) δ ppm 9.43 (d, J=8.8 Hz, 1H), 4.88-4.85 (m, 2H), 4.78-4.68 (m, 2H), 1.47 (s, 9H).
Step c. A mixture of tert-butyl 3-(1-fluoro-2-oxoethylidene)azetidine-1-carboxylate (2.2 g, 10.2 mmol) and ethyl 2-(triphenyl-λ5-phosphanylidene)acetate (CAS: 1099-45-2; 3.56 g, 10.2 mmol) in DCM (100 mL) was degassed and purged with N2 3 times. The reaction mixture was stirred at 25° C. for 12 h under a N2 atmosphere. Upon completion, the mixture was evaporated and purified by column chromatography (PE/EtOAc=5/1) to give tert-butyl (E)-3-(4-ethoxy-1-fluoro-4-oxobut-2-en-1-ylidene)azetidine-1-carboxylate (1.0 g, 34% yield) as a yellow solid.
1H NMR (400 MHz, CDCl3) δ ppm 6.99-6.77 (m, 1H), 6.12 (d, J=15.6 Hz, 1H), 4.64 (d, J=2.8 Hz, 4H), 4.24 (q, J=7.2 Hz, 2H), 1.46 (s, 9H), 1.31 (t, J=7.2 Hz, 3H).
Step d. To a solution of tert-butyl (E)-3-(4-ethoxy-1-fluoro-4-oxobut-2-en-1-ylidene)azetidine-1-carboxylate (1.0 g, 3.50 mmol) in EtOH (20 mL) was added 10% Pd/C (100 mg) under a N2 atmosphere. The suspension was degassed and purged with H2 3 times. The mixture was stirred at 25° C. for 12 h under a H2 atmosphere (15 psi). Upon completion, the reaction mixture was evaporated to give tert-butyl 3-(4-ethoxy-1-fluoro-4-oxobutyl)azetidine-1-carboxylate (1.0 g, 99% yield) as a white oil.
1H NMR (400 MHz, CDCl3) δ ppm 4.76-4.51 (m, 1H), 4.21-4.09 (m, 2H), 4.04-3.94 (m, 2H), 3.93-3.83 (m, 1H), 3.81-3.69 (m, 1H), 2.80-2.61 (m, 1H), 2.56-2.40 (m, 2H), 1.96-1.76 (m, 1H), 1.63-1.53 (m, 1H), 1.44 (s, 9H), 1.32-1.20 (m, 3H).
Step e. This step was conducted in a similar manner to Intermediate B3, step b.
1H NMR (400 MHz, CDCl3) δ ppm 9.83-9.62 (m, 1H), 4.70-4.42 (m, 1H), 3.95-3.87 (m, 2H), 3.85-3.77 (m, 1H), 3.72-3.62 (m, 1H), 3.51-3.38 (m, 1H), 2.66-2.56 (m, 2H), 2.45-2.33 (m, 1H), 1.79-1.69 (m, 1H), 1.37 (s, 9H).
Step a. Allylmagnesium bromide (1 M in Et2O, 40.1 mL) was added to a solution of Intermediate B90 (4 g, 20.1 mmol) in THF (120 mL) at −40° C. The reaction mixture was at 0° C. for 5 h. Upon completion, the reaction mixture was quenched with sat. aq. NH4Cl and extracted with EtOAc (3×50 mL). The combined organic layers were washed with brine, dried over Na2SO4, evaporated and purified by column chromatography (PE/EtOAc=4/1) to give tert-butyl 3-(2-hydroxypent-4-en-1-yl)azetidine-1-carboxylate (800 mg, 16% yield) as a brown oil.
1H NMR (400 MHz, CDCl3) δ ppm 5.91-5.73 (m, 1H), 5.24-5.03 (m, 2H), 4.08-4.00 (m, 2H), 3.69-3.58 (m, 3H), 2.80-2.68 (m, 1H), 2.33-2.25 (m, 1H), 2.20-2.10 (m, 1H), 1.78-1.73 (m, 2H), 1.60-1.55 (m, 1H), 1.44 (s, 9H).
Step b. To a solution of tert-butyl 3-(2-hydroxypent-4-en-1-yl)azetidine-1-carboxylate (0.8 g, 3.32 mmol) in DCM (20 mL) was added DAST (641 mg, 3.98 mmol) at −60° C. The reaction mixture was stirred at −60° C. for 5 h. Upon completion, the mixture was quenched with sat. aq. NaHCO3 and extracted with EtOAc (3×10 mL). The combined organic layers were washed with brine (10 mL), dried over Na2SO4, evaporated and purified by column chromatography (PE/EtOAc=20/1) to give tert-butyl 3-(2-fluoropent-4-en-1-yl)azetidine-1-carboxylate (400 mg, 49% yield) as a light yellow oil.
1H NMR (400 MHz, CDCl3) δ ppm 5.84-5.76 (m, 1H), 5.17-5.12 (m, 2H), 4.60-4.40 (m, 1H), 4.08-4.03 (m, 2H), 3.62-3.58 (m, 2H), 2.75-2.73 (m, 1H), 2.44-2.35 (m, 2H), 2.00-1.89 (m, 2H), 1.44 (s, 9H).
Step c. This step was conducted in a similar manner to Intermediate B49, step h.
1H NMR (400 MHz, CDCl3) δ ppm 9.80 (s, 1H), 5.10-4.90 (m, 1H), 4.18-3.96 (m, 2H), 3.69-3.51 (m, 2H), 2.96-2.57 (m, 3H), 2.18-1.67 (m, 2H), 1.44 (s, 9H).
Steps a-c. These 3 steps were conducted in a similar manner to Intermediate B92, steps c-e, using Intermediate B90 in step a.
1H NMR (400 MHz, CDCl3) δ ppm 9.78 (t, J=1.6 Hz, 1H), 4.01 (t, J=8.4 Hz, 2H), 3.54-3.51 (m, 2H), 2.52-2.46 (m, 3H), 1.65-1.55 (m, 4H), 1.44 (s, 9H).
Step d. A mixture of tert-butyl 3-(4-oxobutyl)azetidine-1-carboxylate (400 mg, 1.76 mmol) and proline (61 mg, 0.53 mmol) in MeCN (1 mL) was added Selectfluor (935 mg, 2.64 mmol) and TFA (60 mg, 0.53 mmol). The reaction mixture was stirred for 12 h under a N2 atmosphere. Upon completion, the reaction mixture was diluted with H2O (10 mL) and extracted with EtOAc (3×10 mL). The combined organic layers were washed with brine (15 mL), dried over Na2SO4, evaporated and purified by column chromatography (PE/EtOAc=1/1) to give the title compound (130 mg, crude) as a colourless oil.
1H NMR 1H NMR (400 MHz, CDCl3) δ ppm 9.80 (s, 1H), 4.04-4.00 (m, 1H), 3.58-3.54 (m, 2H), 2.52-2.45 (m, 2H), 2.40-2.36 (m, 1H), 1.69-1.61 (m, 4H), 1.45 (s, 9H).
Step a. A mixture of 4-tert-butoxycarbonylmorpholine-3-carboxylic acid (1 g, 4.32 mmol), 4-bromobenzaldehyde (1.20 g, 6.49 mmol), Ir[dF(CF3)ppy]2(dtbpy)(PF6) (97 mg, 0.087 mmol), phthalimide (636 mg, 4.32 mmol), [4,4′-bis(1,1-dimethylethyl)-2,2′-bipyridine]nickel (II) dichloride (86 mg, 0.22 mmol) and 2-(tert-butyl)-1,1,3,3-tetramethylguanidine in DMSO (10 mL) was degassed and purged with N2 for 5 min. The reaction was stirred under irradiation from a 34 W 455 nm LED lamp (7 cm away) at 25° C. (fan cooling) for 14 hr. Upon completion, the reaction mixture was diluted with H2O (30 mL) and extracted with EtOAc (2×25 mL). The combined organic layers were washed with brine (30 mL), dried over Na2SO4, evaporated and purified by column chromatography (PE/EtOAc=5/1) to give the title compound (0.6 g, crude) as a colourless oil.
m/z ES+ [M-Boc+H]+ 192.2; 1H NMR (400 MHz, CDCl3) δ ppm 10.02 (s, 1H), 7.88 (d, J=8.4 Hz, 2H), 7.63 (d, J=8.4 Hz, 2H), 5.14 (s, 1H), 4.36 (d, J=12.0 Hz, 1H), 3.97-3.88 (m, 2H), 3.84 (d, J=13.2 Hz, 1H), 3.68-3.56 (m, 1H), 3.17-3.07 (m, 1H), 1.48 (s, 9H).
Step a. This step was conducted in a similar manner to Intermediate B95, step a, using 4-(tert-butoxycarbonyl)morpholine-3-carboxylic acid and methyl 6-bromonicotinate (CAS: 26218-78-0).
m/z ES+ [M+H]+ 323.0; 1H NMR (400 MHz, CDCl3) δ ppm 9.22 (s, 1H), 8.33-8.23 (m, 1H), 7.27-7.25 (m, 1H), 5.13 (s, 1H), 4.77 (d, J=11.2 Hz, 1H), 3.96 (s, 3H), 3.94-3.83 (m, 3H), 3.66-3.54 (m, 1H), 3.34-3.22 (m, 1H), 1.44 (s, 9H).
Step b. This step was conducted in a similar manner to Intermediate B3, step b.
m/z ES+ [M-tBu+H]+ 237.0; 1H NMR (400 MHz, CDCl3) δ ppm 10.12 (s, 1H), 9.07 (d, J=1.6 Hz, 1H), 8.20-8.13 (m, 1H), 7.36 (d, J=8.0 Hz, 1H), 5.15 (s, 1H), 4.82-4.72 (m, 1H), 3.99-3.82 (m, 3H), 3.67-3.56 (m, 1H), 3.35-3.24 (m, 1H), 1.46 (s, 9H).
Step a. To a mixture of tert-butyl 6-oxa-3-azabicyclo[3.1.0]hexane-3-carboxylate (CAS: 114214-49-2; 2.00 g, 10.80 mmol) and ethyl 1H-pyrazole-4-carboxylate (CAS: 37622-90-5; 1.51 g, 10.8 mmol) in DMF (20 mL) was added Cs2CO3 (7.04 g, 21.6 mmol). The reaction mixture was stirred at 60° C. for 12 h. Upon completion, the mixture was quenched with water (100 mL) and extracted with EtOAc (3×100 mL). The combined organic layers were washed with water (120 mL), brine (2×150 mL), dried over Na2SO4, evaporated and purified by reverse phase flash chromatography (water (0.1% NH4OH)/MeCN) to give ethyl 1-((rac-trans)-1-(tert-butoxycarbonyl)-4-hydroxypyrrolidin-3-yl)-1H-pyrazole-4-carboxylate (2.10 g, 60% yield) as a white solid.
m/z ES+ [M+Na]+ 348.3; 1H NMR (400 MHz, CDCl3) δ ppm 7.94 (s, 2H), 4.68-4.63 (m, 2H), 4.30 (q, J=7.2 Hz, 2H), 4.10-3.97 (m, 1H), 3.91-3.71 (m, 2H), 3.46-3.35 (m, 1H), 3.00 (br s, 1H), 1.48 (s, 9H), 1.35 (t, J=7.2 Hz, 3H).
Step b. A solution of ethyl 1-((rac-trans)-1-(tert-butoxycarbonyl)-4-hydroxypyrrolidin-3-yl)-1H-pyrazole-4-carboxylate (2.10 g, 6.45 mmol) in DCM (25 mL) was degassed and purged with N2 3 times, after which DAST (2.08 g, 12.9 mmol) in DCM (5 mL) was added dropwise at −70° C. The reaction mixture was warmed to 25° C. and stirred at that temperature for 16 h. Upon completion, the mixture was quenched with sat. aq. NaHCO3 (20 mL) and extracted with DCM (3×15 mL). The combined organic layers were washed with brine (40 mL), dried over Na2SO4, evaporated and purified by column chromatography (PE/EtOAc=7/1) followed by reverse phase flash chromatography (water (0.1% NH4OH)/MeCN) to give two products. The relative stereochemistry was confirmed by 2D NMR.
Intermediate B97a: ethyl 1-((rac-trans)-1-(tert-butoxycarbonyl)-4-fluoropyrrolidin-3-yl)-1H-pyrazole-4-carboxylate (680 mg, 31% yield) as a white solid.
m/z ES+ [M-tBu+H]+ 272.4; 1H NMR (400 MHz, DMSO-d6) δ ppm 8.48 (s, 1H), 7.94 (s, 1H), 5.47-5.32 (m, 1H), 5.26-5.23 (m, 1H), 4.25-4.19 (m, 2H), 3.85-3.50 (m, 4H), 1.41 (s, 9H), 1.26 (t, J=7.2 Hz, 3H).
Intermediate B97b: ethyl 1-((rac-cis)-1-(tert-butoxycarbonyl)-4-fluoropyrrolidin-3-yl)-1H-pyrazole-4-carboxylate (580 mg, 27% yield) as a white solid.
m/z ES+ [M-tBu+H]+ 272.1; 1H NMR (400 MHz, DMSO-d6) δ ppm 8.49 (s, 1H), 7.93 (s, 1H), 5.52-5.35 (m, 1H), 5.28-5.15 (m, 1H), 4.25-4.19 (m, 2H), 4.00-3.93 (m, 1H), 3.90-3.76 (s, 1H), 3.74-3.58 (m, 2H), 1.43 (s, 9H), 1.27 (t, J=7.2 Hz, 3H).
The title compounds were prepared by chiral SFC separation of Intermediate B97a.
Intermediate B98a:
m/z ES+ [M-tBu+H]+ 272.3; 1H NMR (400 MHz, CDCl3) δ ppm 7.95-7.92 (m, 2H), 5.46-5.19 (m, 1H), 4.99-4.85 (m, 1H), 4.40-4.23 (m, 2H), 4.07-3.65 (m, 4H), 1.50 (s, 9H), 1.27 (t, J=7.2 Hz, 3H).
Intermediate B98b:
m/z ES+ [M-tBu+H]+ 272.4; 1H NMR 400 MHz, CDCl3) δ ppm 7.96-7.92 (m, 2H), 5.47-5.16 (m, 1H), 5.04-4.81 (m, 1H), 4.43-4.24 (m, 2H), 4.04-3.61 (m, 4H), 1.50 (s, 9H), 1.27 (t, J=7.2 Hz, 3H).
The title compounds were prepared by chiral SFC separation of Intermediate B97b.
Intermediate B98c:
m/z ES+ [M-tBu+H]+ 272.3; 1H NMR (400 MHz, CDCl3) δ ppm 8.08 (d, J=9.2 Hz, 1H), 7.96 (s, 1H), 5.39-5.20 (m, 1H), 5.05-4.88 (m, 1H), 4.36-4.28 (m, 2H), 4.19-4.07 (m, 1H), 3.99-3.61 (m, 3H), 1.50 (s, 9H), 1.36 (t, J=7.2 Hz, 3H).
Intermediate B98d:
m/z ES+ [M-tBu+H]+ 272.3; 1H NMR (400 MHz, CDCl3) δ ppm 8.08 (d, J=9.2 Hz, 1H), 7.96 (s, 1H), 5.40-5.19 (m, 1H), 5.05-4.89 (m, 1H), 4.35-4.26 (m, 2H), 4.19-4.07 (m, 1H), 4.00-3.60 (m, 3H), 1.50 (s, 9H), 1.35 (t, J=7.2 Hz, 3H).
Steps a-b. These 2 steps were conducted in a similar manner to Intermediate B10, steps b-c, using Intermediate B98a in step a.
m/z ES+ [M-tBu+H]+ 228.1; 1H NMR (400 MHz, CDCl3) δ ppm 9.88 (s, 1H), 8.01 (d, J=6.8 Hz, 2H), 5.47-5.22 (m, 1H), 5.04-4.90 (m, 1H), 4.07-3.69 (m, 4H), 1.50 (s, 9H).
The title compound was prepared in a similar manner to Intermediate B99a, using Intermediate B98b in step a.
1H NMR (400 MHz, CDCl3) δ ppm 9.88 (s, 1H), 8.01 (d, J=7.6 Hz, 2H), 5.46-5.21 (m, 1H), 5.05-4.90 (m, 1H), 4.07-3.63 (m, 4H), 1.50 (s, 9H).
The title compound was prepared in a similar manner to Intermediate B99a, using Intermediate B98c in step a.
m/z ES+ [M-tBu+H]+ 228.4; 1H NMR (400 MHz, CDCl3) δ ppm 9.90 (s, 1H), 8.13 (d, J=13.2 Hz, 1H), 8.02 (s, 1H), 5.40-5.23 (m, 1H), 5.08-4.93 (m, 1H), 4.23-4.12 (m, 1H), 4.01-3.63 (m, 3H), 1.50 (s, 9H).
The title compound was prepared in a similar manner to Intermediate B99a, using Intermediate B98d in step a.
m/z ES+ [M-tBu+H]+ 228.3, 1H NMR (400 MHz, CDCl3) δ ppm 9.90 (s, 1H), 8.18-8.09 (m, 1H), 8.02 (s, 1H), 5.40-5.24 (m, 1H), 5.08-4.93 (m, 1H), 4.24-4.09 (m, 1H), 4.01-3.84 (m, 1H), 3.83-3.61 (m, 2H), 1.50 (s, 9H).
The title compound was prepared in a similar manner to Intermediate B97a and Intermediate B97b, using tert-butyl 6-oxa-3-azabicyclo[3.1.0]hexane-3-carboxylate and ethyl 1H-imidazole-4-carboxylate in step a.
m/z ES+ [M+H]+ 328.1; 1H NMR (400 MHz, CDCl3) δ ppm 7.77 (s, 1H), 7.64 (s, 1H), 5.34-5.12 (m, 1H), 4.82-4.63 (m, 1H), 4.43-4.31 (m, 2H), 4.10-4.01 (m, 1H), 3.99-3.80 (m, 1H), 3.79-3.64 (m, 2H), 1.49 (s, 9H), 1.38 (t, J=7.2 Hz, 3H).
The title compounds were prepared by chiral SFC separation of Intermediate B100.
Intermediate B101a:
m/z ES+ [M+H]+ 328.4; 1H NMR (400 MHz, CDCl3) δ ppm 7.77 (s, 1H), 7.67 (s, 1H), 5.33-5.11 (m, 1H), 4.79-4.64 (m, 1H), 4.43-4.32 (m, 2H), 4.17-4.00 (m, 1H), 3.99-3.82 (m, 1H), 3.76-3.65 (m, 2H), 1.50 (s, 9H), 1.39 (t, J=7.2 Hz, 3H).
Intermediate B101b:
m/z ES+ [M+H]+ 328.4; 1H NMR (400 MHz, CDCl3) δ ppm 7.77 (s, 1H), 7.65 (s, 1H), 5.34-5.12 (m, 1H), 4.80-4.63 (m, 1H), 4.41-4.33 (m, 2H), 4.13-4.01 (m, 1H), 3.99-3.80 (m, 1H), 3.76-3.63 (m, 2H), 1.49 (s, 9H), 1.38 (t, J=7.2 Hz, 3H).
The title compound was prepared in a similar manner to Intermediate B99a, using Intermediate B101a in step a.
m/z ES+ [M+H]+ 284.3, 1H NMR (400 MHz, CDCl3) δ ppm 9.90 (s, 1H), 7.80 (s, 1H), 7.73 (s, 1H), 5.34-5.16 (m, 1H), 4.85-4.67 (m, 1H), 4.18-4.03 (m, 1H), 4.01-3.64 (m, 3H), 1.50 (s, 9H).
The title compound was prepared in a similar manner to Intermediate B99a, using Intermediate B101b in step a.
m/z ES+ [M+H]+ 284.2; 1H NMR (400 MHz, CDCl3) δ ppm 9.91 (s, 1H), 7.87-7.69 (m, 2H), 5.35-5.13 (m, 1H), 4.82-4.67 (m, 1H), 4.18-4.04 (m, 1H), 3.94-3.66 (m, 3H), 1.50 (s, 9H).
Step a. A mixture of 1-(tert-butyl) 3-methyl (S)-piperazine-1,3-dicarboxylate (CAS: 314741-39-4; 1.0 g, 4.09 mmol), benzyl (2-bromoethyl)carbamate (CAS: 53844-02-3; 1.27 g, 4.91 mmol), K2CO3 (1.70 g, 12.3 mmol) and potassium iodide (68 mg, 0.41 mmol) in MeCN (20 mL) was stirred at 80° C. for 4 h. Upon completion, the reaction was filtered and evaporated. The crude product was purified by column chromatography (PE/EtOAc=3/1) to give 1-(tert-butyl) 3-methyl (S)-4-(2-(((benzyloxy)carbonyl)amino)ethyl)piperazine-1,3-dicarboxylate (0.9 g, 52% yield) as a colourless oil.
1H NMR (400 MHz, CDCl3) δ ppm 7.47-7.29 (m, 5H), 5.30 (br s, 1H), 5.11 (s, 2H), 3.96-3.35 (m, 7H), 3.32-3.19 (m, 3H), 3.12-3.01 (m, 1H), 2.82-2.70 (m, 1H), 2.65-2.52 (m, 1H), 2.44-2.27 (m, 1H), 1.45 (s, 9H).
Step b. To a solution of 1-(tert-butyl) 3-methyl (S)-4-(2-(((benzyloxy)carbonyl)amino)ethyl)piperazine-1,3-dicarboxylate (900 mg, 2.14 mmol) in MeOH (5 mL) was added 10% Pd/C (100 mg), and the mixture was stirred at 50° C. for 3 h under a H2 atmosphere (15 psi). Upon completion, the reaction was filtered and evaporated to give the title compound (500 mg, crude) as a white solid.
1H NMR (400 MHz, CDCl3) δ ppm 6.03 (br s, 1H), 4.64-4.39 (m, 1H), 4.07-4.03 (m, 1H), 3.69-3.51 (m, 1H), 3.29-3.17 (m, 1H), 3.00-2.87 (m, 2H), 2.86-2.84 (m, 1H), 2.81-2.72 (m, 1H), 2.70-2.63 (m, 1H), 2.60-2.50 (m, 1H), 2.33-2.17 (m, 1H), 1.47 (s, 9H).
The title compound was prepared in a similar manner to Intermediate C1, using 1-(tert-butyl) 3-methyl (R)-piperazine-1,3-dicarboxylate (CAS: 438631-77-7) in step a.
1H NMR (400 MHz, CDCl3) δ ppm 5.75 (br s, 1H), 4.61-4.42 (m, 1H), 4.10-3.98 (m, 1H), 3.64-3.53 (m, 1H), 3.29-3.18 (m, 1H), 3.00-2.73 (m, 4H), 2.71-2.64 (m, 1H), 2.62-2.51 (m, 1H), 2.31-2.20 (m, 1H), 1.46 (s, 9H).
Step a. To a solution of tert-butyl (S)-3-cyanopiperazine-1-carboxylate (CAS: 1217650-60-6; 0.5 g, 2.37 mmol) and Rh(OAc)2 (52 mg, 0.24 mmol) in DCM (10 mL) was added ethyl 2-diazenylacetate (550 mg, 4.7 mmol) in DCM (10 mL) over 30 min at 0° C. The reaction mixture was stirred at 25° C. for 4 d under a N2 atmosphere. Upon completion, the mixture was evaporated and the crude product was purified by column chromatography (PE/EtOAc=2/1) to give tert-butyl (S)-3-cyano-4-(2-ethoxy-2-oxoethyl)piperazine-1-carboxylate (470 mg, 67% yield) as a colourless oil.
1H NMR (400 MHz, CDCl3) δ ppm 4.15-4.08 (m, 3H), 4.00 (s, 2H), 3.28 (d, J=6.0 Hz, 2H), 3.09-2.84 (m, 2H), 2.69-2.67 (m, 1H), 2.59-2.47 (m, 1H), 1.41 (s, 9H), 1.20 (t, J=6.4 Hz, 3H).
Step b. To a solution of tert-butyl (S)-3-cyano-4-(2-ethoxy-2-oxoethyl)piperazine-1-carboxylate (470 mg, 1.58 mmol) in MeOH (80 mL) was added Raney-Ni (0.4 g), and the reaction mixture was stirred at 25° C. for 12 h under a H2 atmosphere (45 psi). Upon completion, the mixture was filtered and evaporated to give the title compound (240 mg, 59% yield) as a white solid.
m/z ES+ [M+H]+ 256.3.
The title compound was prepared in a similar manner to Intermediate C3, using tert-butyl (R)-3-cyanopiperazine-1-carboxylate (CAS: 1217791-74-6) in step a.
1H NMR (400 MHz, CDCl3) δ ppm 5.97 (br s, 1H), 4.09-3.97 (m, 1H), 3.91-3.70 (m, 1H), 3.53-3.48 (m, 1H), 3.32-3.24 (m, 1H), 3.24-3.12 (m, 1H), 3.11-2.99 (m, 1H), 2.94 (d, J=16.4 Hz, 1H), 2.90-2.82 (m, 1H), 2.78-2.57 (m, 1H), 2.53-2.39 (m, 1H), 2.29-2.13 (m, 1H), 1.49 (s, 9H).
Steps a-e. These 5 steps were conducted as described in Intermediate B63, steps a-e.
Step f. A mixture of tert-butyl 3,4-diaminopyrrolidine-1-carboxylate (500 mg, 2.48 mmol) and trimethoxymethane (263 mg, 2.48 mmol) in HFIP (6 mL) was stirred at 34° C. for 12 h. Upon completion, the reaction mixture was evaporated to give tert-butyl 3a,4,6,6a-tetrahydropyrrolo[3,4-d]imidazole-5(1H)-carboxylate (500 mg, crude) as a white oil.
m/z ES+ [M+H]+ 212.1.
Step g. To a mixture of oxalyl chloride (874 mg, 6.89 mmol) in DCM (15 mL) was added DMSO (1.08 g, 13.7 mmol) at −65° C. and the mixture was stirred for 30 min, after which tert-butyl 3a,4,6,6a-tetrahydropyrrolo[3,4-d]imidazole-5(1H)-carboxylate (485 mg, 2.30 mmol) in DCM (3 mL) was added at −65° C. The mixture was stirred for a further 30 min and then TEA (2.09 g, 20.6 mmol) was added and the reaction mixture was allowed to warm to 25° C. over 1 h. Upon completion, the reaction mixture was quenched with H2O (30 mL) and extracted with EtOAc (2×30 mL). The combined organic layers were washed with brine (2×30 mL), dried over Na2SO4 and evaporated. The residue was purified by column chromatography (PE/EtOAc=1/1) to give the title compound (110 mg, 8% yield) as a yellow oil.
m/z ES+ [M+H]+ 210.0.
Step a. To a solution of tert-butyl 3-hydroxyazetidine-1-carboxylate (1.0 g, 5.8 mmol) in THF (15 mL) was added NaH (300 mg, 7.5 mmol, 60% dispersion in mineral oil) at 0° C. The mixture was stirred at rt for 30 min, and then a solution of 2-bromo-6-fluoropyridine (CAS: 144100-07-2; 914 mg, 5.2 mmol) in THF (2 mL) was added dropwise at 0° C. The mixture was stirred at rt for 30 min. Upon completion, the reaction mixture was quenched with sat. aq. NH4Cl (30 mL) at 0° C. and extracted with EtOAc (3×35 mL). The combined organic layers were washed with brine (70 mL), dried over Na2SO4, and evaporated. The crude product was purified by column chromatography (PE/EtOAc=15/1) to give the title compound (720 mg, 38% yield) as a colourless oil.
m/z ES+ [M+H]+ 329.3.
Step a. To a solution of 2-(bromomethyl)acrylic acid (CAS: 72707-66-5; 350 mg, 2.12 mmol) in THF (10 mL) was added Me2NH (2 M in THF, 3.18 mL). The reaction mixture was stirred at 15° C. for 12 h. Upon completion, the mixture was evaporated to give the title compound (300 mg, crude) as a white solid, which was used without further purification.
1H NMR (400 MHz, DMSO-d6) δ ppm 6.11 (s, 1H), 5.66 (s, 1H), 3.47 (s, 2H), 2.55 (s, 3H), 2.50 (s, 3H).
Step a. To a solution of ethyl 2-(hydroxymethyl)acrylate (CAS: 10029-04-6; 1 g, 7.68 mmol) in THF (10 mL) and water (10 mL) was added lithium hydroxide monohydrate (644 mg, 15.3 mmol). The reaction mixture was stirred at 20° C. for 12 h. Upon completion, the reaction mixture was acidified to pH 5 with 0.5 M HCl and extracted with EtOAc (3×20 mL). The combined organic layers were washed with brine (60 mL), dried over Na2SO4 and evaporated to give the title compound (450 mg, crude) as a yellow oil, which was used without further purification.
1H NMR (400 MHz, DMSO-d6) δ ppm 12.50 (br s, 1H), 6.07 (s, 1H), 5.78 (s, 1H), 4.97 (br s, 1H), 4.09 (s, 2H).
Step a. To a solution of ethyl 2-(diethoxyphosphoryl)acetate (CAS: 867-13-0; 10 g, 44.6 mmol) in THF (600 mL) was added KHMDS (1 M in THF, 98.1 mL) dropwise over 10 min at −78° C. under a N2 atmosphere. The reaction mixture was stirred for 30 min at −78° C., after which, a solution of NCS (7.15 g, 53.5 mmol) in THF (200 mL) was added dropwise. The reaction mixture was stirred for 30 min at −78° C. and then warmed to 25° C. and stirred for a further 2 h. Upon completion, the reaction mixture was quenched with 1 M HCl (100 mL) and extracted with EtOAc (2×500 mL). The combined organic layers were washed with brine (500 mL), dried over Na2SO4 and evaporated. The crude product was purified by column chromatography (PE/EtOAc=5/1) to give ethyl 2-chloro-2-(diethoxyphosphoryl)acetate (6.5 g, 56% yield) as a yellow oil.
m/z ES+ [M+H]+ 259.0; 1H NMR (400 MHz, CDCl3) δ ppm 4.51 (d, J=16.4 Hz, 1H) 4.25-4.34 (m, 6H) 1.33-1.40 (m, 9H).
Step b. To a solution of ethyl 2-chloro-2-(diethoxyphosphoryl)acetate (1 g, 3.87 mmol) in THF (20 mL) was added NaH (309 mg, 7.73 mmol, 60% dispersion in mineral oil) at 0° C. The mixture was stirred for 1 h, after which, a solution of tert-butyl methyl(2-oxoethyl)carbamate (CAS: 123387-72-4; 670 mg, 3.87 mmol) in THF (4 mL) was added. The reaction mixture was stirred at 0° C. for a further 1 h. Upon completion, the reaction mixture was quenched with sat. aq. NH4Cl (40 mL) and extracted with EtOAc (3×40 mL). The combined organic layers were washed with brine (40 mL), dried over Na2SO4 and evaporated. The residue was purified by column chromatography (PE/EtOAc=5/1) followed by Prep-TLC (PE/EtOAc=5/1) to give ethyl (Z)-4-((tert-butoxycarbonyl)(methyl)amino)-2-chlorobut-2-enoate (130 mg, 12% yield) as a yellow oil.
m/z ES+ [M+H]+ 278.1; 1H NMR (400 MHz, CDCl3) δ ppm 6.39 (s, 1H) 4.38-4.14 (m, 4H), 2.86 (s, 3H), 1.44 (s, 9H) 1.34 (t, J=7.2 Hz, 3H).
Step c. To a solution of ethyl (Z)-4-((tert-butoxycarbonyl)(methyl)amino)-2-chlorobut-2-enoate (150 mg, 0.54 mmol) in 1,4-dioxane (2 mL) and water (2 mL) was added KOH (151 mg, 2.70 mmol). The reaction mixture was stirred at 60° C. for 2 h. Upon completion, the reaction mixture was acidified to pH 2 with 1 M HCl and extracted with EtOAc (3×20 mL). The combined organic layers were washed with brine (20 mL), dried over Na2SO4 and evaporated to give the title compound (0.13 g, 96% yield) as a yellow oil.
1H NMR (400 MHz, CDCl3) δ ppm 6.40 (s, 1H) 4.33-4.10 (m, 2H) 2.93 (s, 3H) 1.47 (s, 9H).
Step a. A mixture of tert-butyl acrylate (10 g, 78.0 mmol), acetaldehyde (50% purity; 13.7 g, 17.5 mL, 156 mmol) and DABCO (1.75 g, 15.6 mmol) in 1,4-dioxane (50 mL) was stirred at 25° C. for 96 h under a N2 atmosphere. Upon completion, the reaction mixture was quenched with water (200 mL) and extracted with EtOAc (3×120 mL). The combined organic layers were washed with brine (100 mL), dried over Na2SO4 and evaporated. The residue was purified by column chromatography (PE/EtOAc=3/1) to give tert-butyl 3-hydroxy-2-methylenebutanoate (12 g, 89% yield) as a colourless oil.
1H NMR (400 MHz, CDCl3) δ ppm 6.11 (d, J=0.4 Hz, 1H), 5.72 (d, J=0.4 Hz, 1H), 4.57 (q, J=6.4 Hz, 1H), 2.43 (br s, 1H), 1.51 (s, 9H), 1.37 (d, J=6.4 Hz, 3H).
Step b. A mixture of tert-butyl 3-hydroxy-2-methylenebutanoate (500 mg, 2.90 mmol) and TFA (6.62 g, 58.0 mmol) in DCM (5 mL) was stirred at 25° C. for 12 h under a N2 atmosphere. Upon completion, the reaction mixture was evaporated. The residue was dissolved in DCM (100 mL), washed with water (3×50 mL), dried over Na2SO4 and evaporated. The residue was purified by Prep-TLC (EtOAc) to give the title compound (100 mg, 29% yield) as a colourless oil.
1H NMR (400 MHz, DMSO-d6) δ ppm 12.5 (br s, 1H), 6.03 (d, J=1.2 Hz, 1H), 5.81 (d, J=1.2 Hz, 1H), 4.95 (d, J=4.4 Hz, 1H), 4.50-4.38 (m, 1H), 1.23-1.15 (m, 3H).
Step a. This step was conducted as described in Intermediate E4, step a.
Step b. Dimethyl sulfide (649 mg, 10.4 mmol) was added dropwise to a solution of NBS (1.71 g, 9.58 mmol) in DCM (3 mL) at 0° C. The mixture was stirred for 10 min, after which, tert-butyl 3-hydroxy-2-methylenebutanoate (1.5 g, 8.71 mmol) was added. The reaction mixture was stirred at 25° C. for 12 h. Upon completion, the reaction mixture was quenched with water (1 mL) and evaporated. The residue was purified by column chromatography (PE/EtOAc=5/1) to give tert-butyl (Z)-2-(bromomethyl)but-2-enoate (1.5 g, 73% yield) as a yellow oil.
1H NMR (400 MHz, CDCl3) δ ppm 6.99 (q, J=7.2 Hz, 1H), 4.23 (s, 2H), 1.91 (d, J=7.2 Hz, 3H), 1.53 (s, 9H).
Step c. A mixture of tert-butyl (Z)-2-(bromomethyl)but-2-enoate (1.4 g, 5.95 mmol) and Me2NH (2 M in THF, 29.7 mL) in THF (5 mL) was stirred at 25° C. for 1 h under a N2 atmosphere. Upon completion, the reaction mixture was evaporated to give tert-butyl (E)-2-((dimethylamino)methyl)but-2-enoate (1.0 g, crude) as a yellow oil.
1H NMR (400 MHz, CDCl3) δ ppm 6.99 (q, J=7.2 Hz, 1H), 3.23 (s, 2H), 2.29 (s, 6H), 1.89 (d, J=7.2 Hz, 3H), 1.50 (s, 9H).
Step d. A mixture of tert-butyl (E)-2-((dimethylamino)methyl)but-2-enoate (1.0 g, 5.02 mmol) and 1 M HCl (25 mL) was stirred at 100° C. for 15 min under a N2 atmosphere. Upon completion, the reaction mixture was evaporated to give the title compound (1.5 g, crude) as a yellow solid.
1H NMR (400 MHz, D2O) δ ppm 7.35 (q, J=7.2 Hz, 1H), 3.91 (s, 2H), 2.75 (s, 6H), 1.85 (d, J=7.2 Hz, 3H).
Step a. A mixture of Intermediate E5 (100 mg, 0.70 mmol), SOCl2 (249 mg, 2.10 mmol) in DCE (1 mL) was degassed and purged with N2 3 times. The reaction mixture was stirred at 60° C. for 2 h under a N2 atmosphere. Upon completion, the reaction mixture was evaporated to give the title compound (120 mg, crude) as a colourless oil.
1H NMR (400 MHz, DMSO-d6) δ ppm 8.70 (br s, 1H), 7.21 (q, J=7.2 Hz, 1H), 3.88 (d, J=5.6 Hz, 2H), 2.65 (d, J=4.8 Hz, 6H), 1.91 (d, J=7.2 Hz, 3H).
The title compound was prepared in a similar manner to Intermediate E1, using diethyl amine (CAS: 109-89-7).
m/z ES+ [M+H]+ 158.1; 1H NMR (400 MHz, DMSO-d6) δ ppm 9.00 (br s, 1H), 6.53 (s, 1H), 6.29 (s, 1H), 3.94 (s, 2H), 3.13-3.07 (m, 4H), 1.23-1.16 (m, 6H).
The title compound was prepared in a similar manner to Intermediate E1, using 1-methylpiperazine.
1H NMR (400 MHz, CD3OD) δ ppm 6.19 (d, J=1.6 Hz, 1H), 5.70 (d, J=1.6 Hz, 1H), 3.54 (s, 2H), 2.83-2.74 (m, 8H), 2.65 (s, 3H).
Step a. A mixture of 1-ethoxy-2,2-difluoroethan-1-ol (CAS: 148992-43-2; 2 g, 15.9 mmol), ethyl 2-(triphenyl-λ5-phosphanylidene)acetate (CAS: 1099-45-2; 6.63 g, 19.0 mmol) and 4-methylbenzenesulfonic acid monohydrate (362 mg, 1.90 mmol) in toluene (32 mL) was stirred at 120° C. for 16 h. Upon completion, the crude reaction mixture was used directly in the next step without further purification.
Step b. To a solution of ethyl (E)-4,4-difluorobut-2-enoate in toluene (2.4 g, 16.0 mmol, crude), THF (8 mL) and 4 M aq. NaOH (8.0 mL) were added. The reaction mixture was stirred at 20° C. for 1 h. Upon completion, the reaction mixture was washed with EtOAc (30 mL) and the organic layer was discarded. The aqueous layer was acidified to pH 3-4 with 1 M HCl and extracted with EtOAc (3×20 mL). The combined organic layers were dried over Na2SO4, evaporated and purified by column chromatography (PE/EtOAc=3/1) to give the title compound (1.3 g, 67% yield) as a yellow oil.
1H NMR (400 MHz, DMSO-d6) δ ppm 13.06 (br s, 1H), 7.33-6.96 (m, 1H), 6.75-6.67 (m, 1H), 6.36-6.32 (m, 1H).
Step a. A mixture of methyl (E)-4-bromobut-2-enoate (CAS: 1117-71-1; 1.38 g, 6.14 mmol), tert-butyl piperazine-1-carboxylate (1.04 g, 5.59 mmol) and DIPEA (1.80 g, 13.97 mmol) in DCM (10 mL) was stirred at 20° C. for 12 h under a N2 atmosphere. Upon completion, the reaction mixture was quenched with water (80 mL) and extracted with EtOAc (70 mL). The combined organic layers were washed with brine (70 mL), dried over Na2SO4, evaporated and purified by column chromatography (PE/EtOAc=2/1) to give tert-butyl (E)-4-(4-methoxy-4-oxobut-2-en-1-yl)piperazine-1-carboxylate (1.0 g, 57% yield) as a white solid.
m/z ES+ [M+H]+ 285.0; 1H NMR (400 MHz, CDCl3) δ ppm 6.90-6.71 (m, 1H), 5.95-5.78 (m, 1H), 3.67-3.56 (m, 3H), 3.33-3.30 (m, 4H), 3.07-2.96 (m, 2H), 2.30-2.26 (m, 4H), 1.47 (s, 9H).
Step b. This step was conducted in a similar manner to Intermediate E2, step a.
m/z ES− [M−H]− 269.0; 1H NMR (400 MHz, CDCl3) δ ppm 6.79-6.65 (m, 1H), 5.98-5.86 (m, 1H), 3.51-3.40 (m, 4H), 3.19-3.10 (m, 2H), 2.41-2.31 (m, 4H), 1.48 (s, 9H).
Step a. A mixture of tert-butyl (R)-2-cyanopyrrolidine-1-carboxylate (CAS: 228244-20-0; 500 mg, 2.55 mmol), hydroxylamine hydrochloride (266 mg, 3.82 mmol) and Na2CO3 (405 mg, 3.82 mmol) in EtOH (5 mL) was stirred at 60° C. for 90 min. Upon completion, the mixture was diluted with EtOAc (20 mL), filtered and the filtrate was evaporated to give the title compound (0.65 g, crude) as a white solid.
m/z ES+ [M+H]+ 230.1; 1H NMR (400 MHz, CDCl3) δ ppm 5.30 (br s, 1H), 4.75-4.07 (m, 2H), 3.54-3.33 (m, 2H), 2.41-2.23 (m, 1H), 2.20-2.07 (m, 1H), 2.05-1.95 (m, 1H), 1.94-1.79 (m, 2H), 1.47 (s, 9H).
The title compound was prepared in a similar manner to Intermediate F1, using tert-butyl (S)-2-cyanopyrrolidine-1-carboxylate (CAS: 228244-04-0).
m/z ES+ [M+H]+ 230.1; 1H NMR (400 MHz, CDCl3) δ ppm 5.35 (br s, 1H), 5.03-4.21 (m, 2H), 3.57-3.29 (m, 2H), 2.39-2.11 (m, 2H), 2.05-1.82 (m, 3H), 1.47 (s, 9H).
Step a. To a solution of 1-(tert-butyl) 2-methyl (R)-pyrrolidine-1,2-dicarboxylate (CAS: 73323-65-6; 1 g, 4.36 mmol) in water (15 mL) was added hydrazine monohydrate (3.85 g, 65.4 mmol). The reaction mixture was stirred at 100° C. for 3 h. Upon completion, the mixture was extracted with DCM (3×15 mL) and the combined organic layers were dried over Na2SO4, filtered and evaporated to give the title compound (350 mg, 35% yield) as a colourless oil.
m/z ES+ [M+Na]+ 252.1; 1H NMR (400 MHz, DMSO-d6) δ ppm 9.02 (br s, 1H), 4.16 (s, 2H), 4.06-3.90 (m, 1H), 3.31-3.22 (m, 2H), 2.09-1.95 (m, 1H), 1.90-1.63 (m, 3H), 1.44-1.28 (m, 9H).
The title compound was prepared in a similar manner to Intermediate G1, using 1-(tert-butyl) 2-methyl (S)-pyrrolidine-1,2-dicarboxylate (CAS: 59936-29-7).
m/z ES+ [M+Na]+ 252.1; 1H NMR (400 MHz, DMSO-d6) δ ppm 9.02 (br s, 1H), 4.18-4.09 (m, 2H), 4.06-3.92 (m, 1H), 3.35-3.22 (m, 2H), 2.09-1.99 (m, 1H), 1.89-1.78 (m, 1H), 1.77-1.70 (m, 2H), 1.41-1.29 (m, 9H).
The title compound was prepared in a similar manner to Intermediate G1, using 1-(tert-butyl) 2-methyl (R)-azetidine-1,2-dicarboxylate (CAS: 1260593-39-2).
m/z ES+ [M+Na]+ 238.2; 1H NMR (400 MHz, DMSO-d6) δ ppm 9.21 (br s, 1H), 4.50-4.42 (m, 1H), 4.34 (s, 2H), 3.94-3.84 (m, 1H), 3.80 (br s, 1H), 2.45-2.31 (m, 1H), 2.08-1.99 (m, 1H), 1.40 (s, 9H)
The title compound was prepared in a similar manner to Intermediate G1, using 1-(tert-butyl) 2-methyl (S)-azetidine-1,2-dicarboxylate (CAS: 107020-12-2).
1H NMR (400 MHz, CDCl3) δ ppm 8.58-8.01 (m, 1H), 4.65 (t, J=8.0 Hz, 1H), 3.94-3.87 (m, 2H), 3.84-3.76 (m, 2H), 2.44 (d, J=7.2 Hz, 2H), 1.45 (s, 9H).
Step a. To a solution of Intermediate 7a (5 g, 10.8 mmol) and Intermediate A1 (5.2 g, 21.5 mmol) in MeOH (50 mL) was added 4 Å molecular sieves (5 g) and acetic acid (3.2 g, 53.8 mmol). The mixture was stirred at rt for 30 min, and then NaBH3CN (1.69 g, 26.9 mmol) was added at 0° C. The mixture was stirred at rt for 30 min. Upon completion, the mixture was filtered and evaporated under vacuum. Purification by column chromatography (PE/EtOAc=0/1 to DCM/MeOH=15/1) to obtain tert-butyl 3-(4-(2-((3aR,11aS)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)ethyl)piperazin-1-yl)azetidine-1-carboxylate (5 g, 65% yield) as a white solid.
m/z ES+ [M+H]+ 690.3; 1H NMR (400 MHz, CDCl3) δ ppm 8.31 (s, 1H), 7.46-7.38 (m, 1H), 7.29-7.27 (m, 1H), 7.23-7.18 (m, 1H), 7.08 (s, 1H), 4.61 (d, J=8.8 Hz, 1H), 3.96-3.93 (m, 2H), 3.79-3.74 (m, 2H), 3.71-3.65 (m, 1H), 3.62-3.55 (m, 1H), 3.51-3.50 (m, 1H), 3.49 (s, 4H), 3.43 (s, 3H), 3.25-3.22 (m, 2H), 3.19-3.18 (m, 1H), 3.12-3.11 (m, 1H), 2.98-2.92 (m, 2H), 2.90-2.82 (m, 2H), 2.61-2.60 (m, 2H), 2.51 (s, 3H), 2.27-2.23 (m, 1H), 1.42 (s, 9H).
Step b. To a solution of tert-butyl 3-(4-(2-((3aR,11aS)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)ethyl)piperazin-1-yl)azetidine-1-carboxylate (3 g, 4.35 mmol) in DCM (40 mL) was added TFA (15.4 g, 135 mmol). The mixture was stirred at 20° C. for 1 h. Upon completion, the mixture was evaporated under vacuum to obtain (3aR,11aS)-5-(2-(4-(azetidin-3-yl)piperazin-1-yl)ethyl)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione as a TFA salt (2.55 g, crude) as a brown gum.
m/z ES+ [M+H]+ 590.1.
Step c. To a solution of (3aR,11aS)-5-(2-(4-(azetidin-3-yl)piperazin-1-yl)ethyl)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (TFA salt, 2.55 g, 4.32 mmol) in DCM (25 mL) was added DIPEA (2.79 g, 21.62 mmol) and subsequently cooled to 0° C. Acryloyl chloride (CAS: 814-68-6; 705 mg, 7.78 mmol) in DCM (2 mL) was then added dropwise. The mixture was stirred at 0° C. for 10 min. Upon completion, the mixture was diluted with water (100 mL) and basified to pH 8 with sat. aq. NaHCO3. The organic layer was separated and the aqueous layer was extracted with DCM (2×150 mL). The combined organic layers were washed with brine (200 mL), dried over Na2SO4 and evaporated. The crude product was purified by column chromatography (DCM/MeOH=10/1) to give the title compound (2.14 g, 77% yield) as a yellow solid.
m/z ES+ [M+H]+ 644.3; 1H NMR (400 MHz, CDCl3) δ ppm 8.32 (s, 1H), 7.35-7.32 (m, 1H), 7.22 (d, J=8.0 Hz, 1H), 7.17-7.11 (m, 1H), 7.06 (s, 1H), 6.36-6.29 (m, 1H), 6.22-6.12 (m, 1H), 5.66 (dd, J=1.6, 10.0 Hz, 1H), 4.60 (d, J=8.8 Hz, 1H), 4.23-4.17 (m, 1H), 4.11-4.02 (m, 2H), 3.91 (dd, J=5.6, 10.6 Hz, 1H), 3.61-3.51 (m, 1H), 3.35 (s, 3H), 3.31-3.23 (m, 1H), 3.22-3.13 (m, 2H), 2.95-2.86 (m, 2H), 2.73-2.68 (m, 1H), 2.61-2.13 (m, 14H).
The Examples in Table 1 were prepared using methods similar to those described in the synthesis of Example 1, using the listed Intermediates in step a.
The Examples in Table 2 were prepared using methods similar to those described in the synthesis of Example 1, using the listed Intermediates in step a, and an additional chiral SFC step conducted after step a.
1H NMR (400 MHz) δ ppm
1H NMR (400 MHz) δ ppm
Steps a-b. These 2 steps were conducted in a similar manner to Example 1, steps a-b, using Intermediate 7a and tert-butyl 5-oxa-2,8-diazaspiro[3.5]nonane-2-carboxylate (CAS: 1251011-05-8) in step a.
Step c. A mixture of (3aR,11aS)-5-(2-(5-oxa-2,8-diazaspiro[3.5]nonan-8-yl)ethyl)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (40 mg, 0.069 mmol), (E)-4-(dimethylamino)but-2-enoic acid hydrochloride (CAS: 848133-35-7; 14 mg, 0.083 mmol), HATU (26 mg, 0.069 mmol) and DIPEA (9 mg, 0.069 mmol) in DCM (2 mL) was stirred at 25° C. for 1 h under a N2 atmosphere. Upon completion, the mixture was quenched with water (0.1 mL) and evaporated. The residue was purified by Prep-HPLC. The combined product containing fractions were evaporated, and the residue was lyophilised with 0.1 N hydrochloric acid to give the title compound as a HCl salt (22 mg, 43% yield) as a white solid.
m/z ES+ [M+H]+ 688.5; 1H NMR (400 MHz, CD3OD) δ ppm 8.32 (s, 1H), 7.56-7.44 (m, 2H), 7.31 (t, J=9.2 Hz, 1H), 7.26 (s, 1H), 6.81-6.70 (m, 1H), 6.54 (d, J=16.0 Hz, 1H), 4.70 (d, J=9.6 Hz, 1H), 4.60-4.44 (m, 1H), 4.32 (d, J=8.8 Hz, 1H), 4.21-4.13 (m, 1H), 4.05-3.96 (m, 5H), 3.77-3.66 (m, 2H), 3.60-3.50 (m, 2H), 3.42 (d, J=3.6 Hz, 3H), 3.40-3.38 (m, 2H), 3.29-3.20 (s, 3H), 3.04 (t, J=12.4 Hz, 1H), 2.95-2.79 (m, 7H), 2.78-2.67 (m, 1H), 2.57 (s, 3H), 2.45-2.38 (m, 1H).
Step a. To a solution of Intermediate 10a (20 mg, 0.03 mmol) in DCM (1 mL) was added BCl3 (1 M in DCM; 0.26 mL) at 0° C. dropwise. The mixture was stirred at 25° C. for 30 min. Upon completion, the reaction mixture was poured into sat. aq. NaHCO3 (2 mL). The mixture was extracted with EtOAc (3×2 mL). The combined organic layers were dried over Na2SO4 and evaporated to give (3aR,11aS)-5-(2-((R)-4-(azetidin-3-yl)-3-(fluoromethyl)piperazin-1-yl)ethyl)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (16 mg, crude) as a yellow solid.
m/z ES+ [M+H]+ 622.6.
Step b. To a solution of (3aR,11aS)-5-(2-((R)-4-(azetidin-3-yl)-3-(fluoromethyl)piperazin-1-yl)ethyl)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (15 mg, 0.02 mmol) and DIPEA (6 mg, 0.05 mmol) in DCM (0.5 mL) was added a solution of acryloyl chloride (2 mg, 0.02 mmol) in DCM (0.1 mL) at 0° C. The mixture was stirred at 0° C. for 10 min. Upon completion, the reaction mixture was evaporated. The residue was purified by Prep-HPLC to give the title compound (4 mg, 23% yield) as a white solid.
m/z ES+ [M+H]+ 676.3; 1H NMR (400 MHz, CDCl3) δ ppm 8.33 (s, 1H), 7.41-7.33 (m, 1H), 7.27-7.23 (m, 1H), 7.21-7.13 (m, 1H), 7.06 (s, 1H), 6.37-6.29 (m, 1H), 6.22-6.12 (m, 1H), 5.70-5.66 (m, 1H), 4.79-4.31 (m, 3H), 4.30-4.05 (m, 3H), 4.03-3.91 (m, 1H), 3.82-3.54 (m, 3H), 3.39 (s, 5H), 3.18-2.53 (m, 11H), 2.50 (s, 3H), 2.25-2.20 (m, 1H).
The title compound was prepared in a similar manner to Example 28, using Intermediate 10b in step a.
m/z ES+ [M+H]+ 676.3; 1H NMR (400 MHz, CDCl3) δ ppm 8.32 (s, 1H), 7.40-7.32 (m, 1H), 7.23 (d, J=8.0 Hz, 1H), 7.19-7.12 (m, 1H), 7.06 (s, 1H), 6.38-6.30 (m, 1H), 6.23-6.13 (m, 1H), 5.71-5.64 (m, 1H), 4.60 (m, 1H), 4.23 (m, 1H), 4.14-4.06 (m, 1H), 4.04-3.93 (m, 1H), 3.83 (m, 1H), 3.64-3.56 (m, 1H), 3.55-3.46 (m, 1H), 3.44-3.12 (m, 6H), 3.09-2.51 (m, 12H), 2.50 (s, 4H), 2.20 (m, 1H).
Step a. To a solution of Intermediate 7a (80 mg, 0.17 mmol) and Intermediate A4 (62 mg, 0.17 mmol) in MeOH (1 mL) was added 4 Å molecular sieves (10 mg) and acetic acid (1 mg, 0.02 mmol). The mixture was stirred at rt for 5 min, and then NaBH3CN (13 mg, 0.21 mmol) was added. The mixture was stirred at rt for a further 25 min. Upon completion, the mixture was diluted with sat. aq. NaHCO3 (5 mL), and then extracted with EtOAc (3×5 mL). The combined organic layers were washed with brine (10 mL), dried over Na2SO4 and evaporated. The crude product was purified by Prep-TLC (PE/EtOAc=1/1) to give 5-benzyl 2-(tert-butyl) 8-(2-((3aR,11aS)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)ethyl)-2,5,8-triazaspiro[3.5]nonane-2,5-dicarboxylate (80 mg, 45% yield) as a yellow solid.
m/z ES+ [M+H]+ 810.3.
Step b. To a solution of 5-benzyl 2-(tert-butyl) 8-(2-((3aR,11aS)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)ethyl)-2,5,8-triazaspiro[3.5]nonane-2,5-dicarboxylate (70 mg, 0.07 mmol) in DCM (1 mL) was added TFA (308 mg, 2.70 mmol). The mixture was stirred at rt for 30 min. Upon completion, the mixture was diluted with DCM (3 mL) and then basified to pH 8 with sat. aq. NaHCO3. The organic layer was separated and the aqueous was further extracted with DCM (2×4 mL). The combined organic layers were washed with brine (6 mL), dried over Na2SO4 and evaporated to give benzyl 8-(2-((3aR,11aS)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)ethyl)-2,5,8-triazaspiro[3.5]nonane-5-carboxylate as a TFA salt (60 mg, crude) as a yellow oil.
m/z ES+ [M+H]+ 710.2.
Step c. To a solution of benzyl 8-(2-((3aR,11aS)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)ethyl)-2,5,8-triazaspiro[3.5]nonane-5-carboxylate (60 mg, 0.08 mmol) in DCM (0.5 mL) was added DIPEA (22 mg, 0.17 mmol)) and subsequently cooled to 0° C. Acryloyl chloride (8 mg, 0.09 mmol) in DCM (0.1 mL) was added dropwise. The mixture was stirred at 0° C. for 10 min. Upon completion, the reaction mixture was evaporated under vacuum. The crude product was purified by reverse phase flash chromatography (water (0.1% NH4OH)/MeCN) to give benzyl 2-acryloyl-8-(2-((3aR,11aS)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)ethyl)-2,5,8-triazaspiro[3.5]nonane-5-carboxylate (40 mg, 55% yield) as a white solid.
m/z ES+ [M+H]+ 764.3; 1H NMR (400 MHz, CDCl3) δ ppm 8.33 (s, 1H), 7.36-7.34 (m, 6H), 7.23-7.22 (m, 1H), 7.15 (t, J=8.8 Hz, 1H), 7.06 (s, 1H), 6.39-6.31 (m, 1H), 5.70 (d, J=9.6 Hz, 1H), 5.31 (s, 2H), 5.20-5.10 (m, 2H), 4.59 (d, J=8.8 Hz, 1H), 4.26-3.67 (m, 5H), 3.61-3.40 (m, 3H), 3.35 (s, 3H), 3.33-3.12 (m, 2H), 2.99-2.66 (m, 5H), 2.49 (s, 3H), 2.44-2.16 (m, 3H).
Step d. A mixture of benzyl 2-acryloyl-8-(2-((3aR,11aS)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)ethyl)-2,5,8-triazaspiro[3.5]nonane-5-carboxylate (40 mg, 0.051 mmol) and TFA (0.5 mL) was stirred at 45° C. for 16 h. Upon completion, the mixture was evaporated under vacuum, and the residue was dissolved in DCM (5 mL) and basified to pH 9 with sat. aq. Na2CO3. The organic layer was separated and the aqueous was extracted with DCM (2×5 mL). The combined organic layers were washed with brine (10 mL), dried over Na2SO4 and evaporated. The crude product was purified by Prep-HPLC to give the title compound (11 mg, 35% yield) as a yellow solid.
m/z ES+ [M+H]+ 630.3; 1H NMR (400 MHz, CDCl3) δ ppm 8.33 (s, 1H), 7.35-7.33 (m, 1H), 7.22 (d, J=7.6 Hz, 1H), 7.14 (t, J=9.2 Hz, 1H), 7.06 (s, 1H), 6.40-6.31 (m, 1H), 6.27-6.16 (m, 1H), 5.72-5.66 (m, 1H), 4.61 (d, J=9.2 Hz, 1H), 4.04 (d, J=8.4 Hz, 1H), 3.92-3.84 (m, 2H), 3.72 (d, J=10.8 Hz, 1H), 3.60-3.52 (m, 1H), 3.35 (d, J=2.0 Hz, 3H), 3.24-3.16 (m, 2H), 2.98-2.78 (m, 4H), 2.72-2.68 (m, 1H), 2.61-2.50 (m, 1H), 2.49 (s, 3H), 2.48-2.37 (m, 3H), 2.37-2.26 (m, 2H), 2.21 (m, 1H).
The Examples in Table 3 were prepared using methods similar to those described in the synthesis of Example 30, using the listed Intermediates in step a.
The Examples in Table 4 were prepared using methods similar to those described in the synthesis of Example 30, using the listed Intermediates in step a, and an additional chiral SFC step was conducted after step a.
1H NMR (400 MHz) δ ppm
1H NMR (400 MHz) δ ppm
Step a. This step was conducted as described in Example 30, step a, using Intermediate 7a and Intermediate A4.
Step b. To a solution of 5-benzyl 2-(tert-butyl) 8-(2-((3aR,11aS)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)ethyl)-2,5,8-triazaspiro[3.5]nonane-2,5-dicarboxylate (110 mg, 0.14 mmol) in THF (4 mL) was added 10% Pd/C (29 mg), and the mixture was stirred at 25° C. for 1 h under a H2 atmosphere (15 psi). Upon completion, the reaction mixture was filtered and evaporated to give tert-butyl 8-(2-((3aR,11aS)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)ethyl)-2,5,8-triazaspiro[3.5]nonane-2-carboxylate (90 mg, 98% yield) as a colourless solid.
m/z ES+ [M+H]+ 676.4.
Step c. To a solution of tert-butyl 8-(2-((3aR,11aS)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)ethyl)-2,5,8-triazaspiro[3.5]nonane-2-carboxylate (90 mg, 0.13 mmol) and acetaldehyde (18 mg, 0.40 mmol) in MeOH (3 mL) was added acetic acid (40 mg, 0.67 mmol), 4 Å molecular sieves (100 mg) and NaBH3CN (17 mg, 0.27 mmol). The mixture was stirred at 25° C. for 1 h. Upon completion, the reaction mixture was filtered and evaporated to give tert-butyl 5-ethyl-8-(2-((3aR,11aS)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)ethyl)-2,5,8-triazaspiro[3.5]nonane-2-carboxylate (90 mg, crude) as a colourless solid.
m/z ES+ [M+H]+ 704.4.
Step d. To a solution of tert-butyl 5-ethyl-8-(2-((3aR,11aS)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)ethyl)-2,5,8-triazaspiro[3.5]nonane-2-carboxylate (90 mg, 0.13 mmol) in DCM (3 mL) was added TFA (1.46 g, 12.8 mmol) and the mixture was stirred at 25° C. for 30 min. Upon completion, the reaction mixture was evaporated to give (3aR,11aS)-5-(2-(5-ethyl-2,5,8-triazaspiro[3.5]nonan-8-yl)ethyl)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione as a TFA salt (90 mg, crude) as a yellow solid.
m/z ES+ [M+H]+ 604.4.
Step e. To a solution of (3aR,11aS)-5-(2-(5-ethyl-2,5,8-triazaspiro[3.5]nonan-8-yl)ethyl)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (90 mg, 0.13 mmol) in DCM (3 mL) was added DIPEA (81 mg, 0.63 mmol) and acryloyl chloride (17 mg, 0.19 mmol). The mixture was stirred at 25° C. for 30 min. Upon completion, the reaction mixture was evaporated and the resulting residue was purified by Prep-HPLC to give the title compound (47 mg, 56% yield) as a yellow solid.
m/z ES+ [M+H]+ 658.4; 1H NMR (400 MHz, CDCl3) δ ppm 8.32 (s, 1H), 7.33 (td, J=8.4, 5.6 Hz, 1H), 7.21 (d, J=7.6 Hz, 1H), 7.13 (t, J=9.2 Hz, 1H), 7.05 (s, 1H), 6.38-6.31 (m, 1H), 6.28-6.15 (m, 1H), 5.69-5.65 (m, 1H), 4.60 (d, J=9.2 Hz, 1H), 4.21-4.14 (m, 1H), 4.05 (d, J=11.2 Hz, 1H), 3.78 (d, J=9.2 Hz, 1H), 3.62 (dd, J=10.8, 6.4 Hz, 1H), 3.58-3.52 (m, 1H), 3.34 (d, J=2.0 Hz, 3H), 3.25-3.15 (m, 2H), 2.96-2.82 (m, 2H), 2.70 (dd, J=16.4, 8.0 Hz, 2H), 2.63-2.57 (m, 4H), 2.49 (s, 3H), 2.47-2.39 (m, 3H), 2.37-2.27 (m, 2H), 2.19 (dd, J=16.8, 11.2 Hz, 1H), 1.08 (t, J=7.2 Hz, 3H).
Steps a-b. These steps were conducted as described in Example 35, steps a-b, using Intermediate 7a and Intermediate A4.
Step c. To a solution of tert-butyl 8-(2-((3aR,11aS)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)ethyl)-2,5,8-triazaspiro[3.5]nonane-2-carboxylate (150 mg, 0.22 mmol) and acetic acid (17 mg, 0.29 mmol) in DCM (2 mL) was added DIPEA (72 mg, 0.55 mmol) and HATU (101 mg, 0.27 mmol). The mixture was stirred at 25° C. for 30 min. Upon completion, the mixture was diluted with water (5 mL) and extracted with EtOAc (3×5 mL). The combined organic layers were washed with brine (10 mL), dried over Na2SO4 and evaporated. The residue was purified by Prep-TLC (EtOAc) to give tert-butyl 5-acetyl-8-(2-((3aR,11aS)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)ethyl)-2,5,8-triazaspiro[3.5]nonane-2-carboxylate (145 mg, 91% yield) as a white solid.
m/z ES+ [M+H]+ 718.3; 1H NMR (400 MHz, CDCl3) δ ppm 8.33 (s, 1H), 7.38-7.31 (m, 1H), 7.23 (d, J=8.0 Hz, 1H), 7.18-7.12 (m, 1H), 7.06 (s, 1H), 4.61 (d, J=9.2 Hz, 1H), 4.11-4.04 (m, 1H), 4.03-3.90 (m, 1H), 3.78 (dd, J=9.2, 16.0 Hz, 2H), 3.60-3.53 (m, 1H), 3.35 (s, 3H), 3.32-3.23 (m, 1H), 3.20 (m, 2H), 2.97-2.85 (m, 2H), 2.81 (s, 4H), 2.72 (dd, J=8.0, 16.8 Hz, 1H), 2.66-2.53 (m, 2H), 2.50 (s, 3H), 2.48-2.37 (m, 2H), 2.36-2.26 (m, 2H), 2.20 (dd, J=11.2, 16.8 Hz, 1H), 1.45 (s, 9H).
Step d. To a solution of tert-butyl 5-acetyl-8-(2-((3aR,11aS)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)ethyl)-2,5,8-triazaspiro[3.5]nonane-2-carboxylate (120 mg, 0.17 mmol) in DCM (2 mL) was added TFA (1 mL). The mixture was stirred at 25° C. for 30 min. Upon completion, the mixture was adjusted to pH 8 with sat. aq. NaHCO3 and then extracted with DCM (3×5 mL). The combined organic layers were washed with brine (10 mL), dried over Na2SO4 and evaporated to give (3aR,11aS)-5-(2-(5-acetyl-2,5,8-triazaspiro[3.5]nonan-8-yl)ethyl)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (100 mg, crude) as a white solid.
m/z ES+ [M+H]+ 618.6.
Step e. To a solution of (3aR,11aS)-5-(2-(5-acetyl-2,5,8-triazaspiro[3.5]nonan-8-yl)ethyl)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (100 mg, 0.16 mmol) in DCM (1 mL) was added DIPEA (42 mg, 0.32 mmol). A solution of acryloyl chloride (19 mg, 0.21 mmol) in DCM (0.1 mL) was added dropwise at 0° C. The mixture was stirred at 0° C. for 10 min. Upon completion, the mixture was diluted with water (10 mL) and extracted with EtOAc (3×10 mL). The combined organic layers were washed with brine (20 mL), dried over Na2SO4 and evaporated. The crude product was purified by Prep-HPLC to give the title compound (37 mg, 31% yield) as a white solid.
m/z ES+ [M+H]+ 672.3; 1H NMR (400 MHz, CDCl3) δ ppm 8.32 (s, 1H), 7.39-7.31 (m, 1H), 7.28-7.23 (m, 1H), 7.18-7.11 (m, 1H), 7.05 (s, 1H), 6.39-6.30 (m, 1H), 6.26-6.13 (m, 1H), 5.69-5.67 (m, 1H), 4.60 (d, J=9.2 Hz, 1H), 4.55-4.40 (m, 1H), 4.15-4.01 (m, 2H), 3.95 (s, 1H), 3.58-3.50 (m, 1H), 3.48-3.36 (m, 1H), 3.34 (d, J=3.2 Hz, 3H), 3.30-3.14 (m, 3H), 2.98-2.79 (m, 2H), 2.77-2.64 (m, 2H), 2.64-2.55 (m, 1H), 2.54-2.38 (m, 5H), 2.36-2.29 (m, 1H), 2.29-2.14 (m, 2H), 2.06 (d, J=2.0 Hz, 3H).
Step a. A mixture of Intermediate 7a (0.3 g, 0.65 mmol), Intermediate A8 (503 mg, 1.29 mmol), acetic acid (193 mg, 3.23 mmol) and 4 Å molecular sieves (0.3 g) in MeOH (3 mL) was stirred at 25° C. for 30 min, after which, NaBH3CN (41 mg, 0.65 mmol) was added and the mixture was stirred at 25° C. for a further 1 h. Upon completion, the mixture was filtered and evaporated. The residue was purified by Prep-HPLC to give 5-benzyl 2-(tert-butyl) (6R)-8-(2-((3aR,11aS)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)ethyl)-6,9-dimethyl-2,5,8-triazaspiro[3.5]nonane-2,5-dicarboxylate (300 mg, 55% yield) as a white solid.
m/z ES+ [M+H]+ 838.6.
Step b. A mixture 5-benzyl 2-(tert-butyl) (6R)-8-(2-((3aR,11aS)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)ethyl)-6,9-dimethyl-2,5,8-triazaspiro[3.5]nonane-2,5-dicarboxylate (100 mg, 0.12 mmol) and 10% Pd/C (20 mg) in 2-propanol (1 mL) was stirred at 25° C. under a H2 atmosphere (15 psi) for 12 h. Upon completion, the mixture was filtered and evaporated to give tert-butyl (6R)-8-(2-((3aR,11aS)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)ethyl)-6,9-dimethyl-2,5,8-triazaspiro[3.5]nonane-2-carboxylate (83 mg, crude) as a white solid.
m/z ES+ [M+H]+ 704.4.
Step c. A mixture of tert-butyl (6R)-8-(2-((3aR,11aS)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)ethyl)-6,9-dimethyl-2,5,8-triazaspiro[3.5]nonane-2-carboxylate (83 mg, 0.12 mmol) and TFA (1 mL) in DCM (1 mL) was stirred at 25° C. for 1 h. Upon completion, the mixture was adjusted pH to 8-9 with sat. aq. NaHCO3 and diluted with water (30 mL). The mixture was extracted with EtOAc (2×30 mL). The combined organic layers were dried over Na2SO4 and evaporated to give (3aR,11aS)-5-(2-((6R)-6,9-dimethyl-2,5,8-triazaspiro[3.5]nonan-8-yl)ethyl)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (60 mg, 84% yield) as a colourless oil.
m/z ES+ [M+H]+ 604.3.
Step d. To a solution of (3aR,11aS)-5-(2-((6R)-6,9-dimethyl-2,5,8-triazaspiro[3.5]nonan-8-yl)ethyl)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (60 mg, 0.099 mmol), acrylic acid (11 mg, 0.15 mmol) and HATU (49 mg, 0.13 mmol) in DCM (1 mL) was added DIPEA (26 mg, 0.20 mmol). The mixture was stirred at 25° C. for 1 h. Upon completion, the mixture was evaporated. The residue was purified by Prep-HPLC to give two diastereoisomers with unknown absolute configuration.
Example 37a: (15 mg, 23% yield) as an off-white solid.
m/z ES+ [M+H]+ 658.3; 1H NMR (400 MHz, CDCl3) δ ppm 8.34 (s, 1H), 7.40-7.32 (m, 1H), 7.26-7.21 (m, 1H), 7.16 (t, J=9.2 Hz, 1H), 7.07 (s, 1H), 6.39-6.31 (m, 1H), 6.28-6.14 (m, 1H), 5.77-5.60 (m, 1H), 4.62 (d, J=8.8 Hz, 1H), 4.10-3.78 (m, 1H), 4.10-3.78 (m, 2H), 3.76-3.48 (m, 3H), 3.37 (s, 3H), 3.26-3.15 (m, 2H), 2.99-2.81 (m, 2H), 2.80-2.60 (m, 3H), 2.51 (s, 3H), 2.41-2.15 (m, 3H), 2.04-1.89 (m, 1H), 1.56 (t, J=7.2 Hz, 1H), 1.16-0.99 (m, 6H).
Example 37b: (15 mg, 23% yield) as a white solid.
m/z ES+ [M+H]+ 658.3; 1H NMR (400 MHz, CDCl3) δ ppm 8.34 (s, 1H), 7.36 (dt, J=5.6, 8.4 Hz, 1H), 7.24 (d, J=8.0 Hz, 1H), 7.16 (t, J=9.2 Hz, 1H), 7.07 (s, 1H), 6.41-6.31 (m, 1H), 6.31-6.13 (m, 1H), 5.74-5.64 (m, 1H), 4.71-4.57 (m, 1H), 4.17-4.01 (m, 1H), 3.98-3.84 (m, 2H), 3.77-3.53 (m, 2H), 3.44-3.30 (m, 3H), 3.26-3.05 (m, 2H), 3.00-2.80 (m, 4H), 2.79-2.67 (m, 1H), 2.51 (s, 3H), 2.47-2.30 (m, 3H), 2.28-2.09 (m, 2H), 1.57 (t, J=7.2 Hz, 1H), 1.12-0.99 (m, 3H), 0.88 (d, J=6.4 Hz, 3H).
Step a. A mixture of Intermediate B1 (140 mg, 0.56 mmol), Intermediate 5a (150 mg, 0.36 mmol) and acetic acid (107 mg, 1.78 mmol) in MeOH (3 mL) was stirred at 25° C. for 30 min, then NaBH3CN (44.6 mg, 0.71 mmol) was added. The reaction mixture was stirred at 25° C. for 12 h. Upon completion, the reaction mixture was filtered and evaporated. The residue was purified by Prep-TLC (EtOAc/MeOH=10/1) to give tert-butyl 3-(4-(((3aR,11aS)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)methyl)-1H-imidazol-1-yl)azetidine-1-carboxylate (100 mg, 40% yield) as a yellow oil.
m/z ES+ [M+H]+ 658.3.
Step b. To a solution of tert-butyl 3-(4-(((3aR,11aS)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)methyl)-1H-imidazol-1-yl)azetidine-1-carboxylate (60 mg, 0.091 mmol) in DCM (3 mL) was added TFA (1 mL). The reaction mixture was stirred at 20° C. for 30 min. Upon completion, the reaction mixture was evaporated. The residue was basified with sat. aq. NaHCO3 (30 mL) and extracted with EtOAc (30 mL). The organic layer was washed with brine (30 mL), dried over Na2SO4 and evaporated to give (3aR,11aS)-5-((1-(azetidin-3-yl)-1H-imidazol-4-yl)methyl)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione as a TFA salt (50 mg, crude) as a yellow gum.
m/z ES+ [M+H]+ 558.2.
Step c. To a solution of (3aR,11aS)-5-[[1-(azetidin-3-yl)imidazol-4-yl]methyl]-6-fluoro-10-methyl-1-[6-methyl-4-(trifluoromethyl)-2-pyridyl]-3,3a,4,11a-tetrahydropyrrolo[2,3-c][1,6]benzodiazocine-2,11-dione (TFA salt, 50 mg, 0.09 mmol) and DIPEA (35 mg, 0.27 mmol) in DCM (2 mL) was added acryloyl chloride (12 mg, 0.14 mmol). The reaction mixture was stirred at 20° C. for 10 min. Upon completion, the reaction mixture was diluted with water (20 mL) and extracted with EtOAc (20 mL). The organic layer was washed with brine (20 mL), dried over Na2SO4 and evaporated. The residue was purified by Prep-HPLC to give the title compound (31 mg, 54% yield) as a white solid.
m/z ES+ [M+H]+ 612.2; 1H NMR (400 MHz, CDCl3) δ ppm 8.86 (s, 1H), 8.30 (s, 1H), 7.42-7.32 (m, 1H), 7.25-7.10 (m, 3H), 7.06 (s, 1H), 6.46-6.30 (m, 1H), 6.27-6.12 (m, 1H), 5.75 (d, J=10.0 Hz, 1H), 5.36-5.34 (m, 1H), 4.76-4.73 (m, 1H), 4.61 (d, J=9.2 Hz, 2H), 4.52-4.76 (m, 2H), 4.26-4.20 (m, 2H), 3.77-3.67 (m, 1H), 3.28 (s, 3H), 3.07-2.84 (m, 2H), 2.82-2.68-2.55 (m, 1H), 2.49 (s, 3H), 2.26-2.10 (m, 1H).
Step a. To a solution of Intermediate 5b (349 mg, 0.80 mmol), Intermediate B1 (300 mg, 1.19 mmol) in DMF (3 mL) was added TMSCI (216 mg, 1.99 mmol). The mixture was stirred at 0° C. for 30 min. A solution of borane tetrahydrofuran complex (1 M in THF, 0.80 mL) in THF (7 mL) was added and the reaction mixture was stirred at 0° C. for 90 min, and then at 20° C. for 14 h. (LCMS showed some Boc-group cleavage). Upon completion, the reaction mixture was basified with NaHCO3 (201 mg, 2.39 mmol), and di-tert-butyl dicarbonate (347 mg, 1.59 mmol) was added. The mixture was stirred at 20° C. for 3 h. Upon completion, the reaction mixture was quenched with methanol (4 mL) at 0° C., filtered and evaporated. The crude product was purified by reverse phase flash chromatography (water (0.1 FA)/MeCN) to give tert-butyl 3-(4-(((3aR,11aS)-6-chloro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)methyl)-1H-imidazol-1-yl)azetidine-1-carboxylate (200 mg, 34% yield) as light yellow oil.
m/z ES+ [M+H]+ 674.2.
Steps b-c. These 2 steps were conducted in a similar manner to Example 38, steps b-c.
m/z ES+ [M+H]+ 628.1; 1H NMR (400 MHz, CDCl3) δ ppm 8.32 (s, 1H), 7.53 (s, 1H), 7.46 (dd, J=2.4, 7.2 Hz, 1H), 7.34-7.28 (m, 2H), 7.13 (s, 1H), 7.05 (s, 1H), 6.45-6.36 (m, 1H), 6.27-6.17 (m, 1H), 5.76 (d, J=10.4 Hz, 1H), 5.00-4.96 (m, 1H), 4.75-4.57 (m, 3H), 4.37-4.34 (m, 1H), 4.33-4.21 (m, 2H), 4.19-4.11 (m, 1H), 3.68-3.63 (m, 1H), 3.35-3.33 (m, 3H), 3.10-2.99 (m, 1H), 2.98-2.83 (m, 1H), 2.74-2.62 (m, 1H), 2.48 (s, 3H), 2.24-2.16 (m, 1H).
The Examples in Table 5 were prepared using methods similar to those described in the synthesis of Example 38 or Example 39, using the listed Intermediates in step a, and the appropriate warhead in step c.
1H NMR (400 MHz) δ ppm
The title compound was prepared in a similar manner to Example 38, using Intermediate 5a and Intermediate B4 in step a. The final step was conducted in a similar manner to the amide coupling described for Example 27.
m/z ES+ [M+H]+ 647.4; 1H NMR (400 MHz, DMSO-d6) δ ppm 8.17 (s, 1H), 7.43-7.20 (m, 4H), 7.00-6.92 (m, 1H), 6.67-6.60 (m, 1H), 6.60-6.54 (m, 1H), 6.08-6.04 (m, 1H), 4.58 (d, J=9.2 Hz, 1H), 4.37-4.30 (m, 1H), 4.26-4.20 (m, 1H), 4.10-3.90 (m, 3H), 3.63-3.58 (m, 1H), 3.50-3.45 (m, 1H), 3.42-3.40 (m, 1H), 3.24 (s, 1H), 3.19-3.14 (m, 1H), 3.13-3.04 (m, 1H), 3.04-2.90 (m, 2H), 2.12 (d, J=2.4 Hz, 2H), 2.04-1.90 (m, 2H), 1.68-1.61 (m, 1H), 1.60-1.42 (m, 2H), 1.41-1.20 (m, 6H), 0.94 (t, J=7.4 Hz, 1H), 0.88-0.83 (m, 1H).
The title compound was prepared in a similar manner to Example 43, using Intermediate 5b and Intermediate B4 in step a.
m/z ES+ [M+H]+ 663.1; 1H NMR (400 MHz, CDCl3) δ ppm 8.33 (s, 1H), 7.49-7.42 (m, 1H), 7.34-7.29 (m, 2H), 7.05 (s, 1H), 6.90-6.76 (m, 1H), 6.41-6.22 (m, 1H), 4.63 (d, J=9.2 Hz, 1H), 4.47-4.35 (m, 1H), 4.32-4.15 (m, 2H), 4.13-3.99 (m, 1H), 3.95-3.78 (m, 1H), 3.59-3.52 (m, 1H), 3.50-3.31 (m, 7H), 3.18-2.93 (m, 4H), 2.74 (dd, J=7.6, 16.8 Hz, 1H), 2.62-2.50 (m, 6H), 2.48 (s, 3H), 2.30-2.17 (m, 1H), 1.75-1.53 (m, 2H).
Step a. To a solution of Intermediate C1 (177 mg, 0.69 mmol) in DMF (5 mL) was added NaH (21 mg, 0.52 mmol, 60% dispersion in mineral oil). The mixture was stirred at 0° C. for 30 min, then Intermediate 8 (200 mg, 0.35 mmol) was added. The mixture was stirred at 20° C. for 30 min. Upon completion, the mixture was poured into sat. aq. NH4Cl (10 mL) at 0° C. and extracted with EtOAc (3×10 mL). The combined organic layers were dried over Na2SO4 and evaporated. The crude product was purified by reverse phase flash chromatography (water (0.1 FA)/MeCN) to give tert-butyl (S)-8-(2-((3aR,11aS)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)ethyl)-9-oxooctahydro-2H-pyrazino[1,2-a]pyrazine-2-carboxylate (80 mg, 33% yield) as a white solid.
1H NMR (400 MHz, CDCl3) δ ppm 8.33 (s, 1H), 7.40-7.31 (m, 1H), 7.23 (d, J=8.0 Hz, 1H), 7.16 (t, J=9.2 Hz, 1H), 7.07 (s, 1H), 4.63 (d, J=9.2 Hz, 1H), 4.58-4.46 (m, 1H), 4.18-3.94 (m, 1H), 3.78-3.54 (m, 2H), 3.49-3.41 (m, 1H), 3.40 (s, 3H), 3.38-3.27 (m, 2H), 3.26-3.16 (m, 2H), 3.01-2.67 (m, 7H), 2.66-2.54 (m, 2H), 2.51 (s, 3H), 2.30-2.13 (m, 2H), 1.47 (s, 9H).
Step b. To a solution of tert-butyl (S)-8-(2-((3aR,11aS)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)ethyl)-9-oxooctahydro-2H-pyrazino[1,2-a]pyrazine-2-carboxylate (60 mg, 0.085 mmol) in DCM (1 mL) was added TFA (770 mg, 6.75 mmol). The mixture was stirred at 15° C. for 30 min. Upon completion, the reaction mixture was evaporated to give (3aR,11aS)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-5-(2-((S)-1-oxooctahydro-2H-pyrazino[1,2-a]pyrazin-2-yl)ethyl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione as a TFA salt (60 mg, crude) as white solid.
m/z ES+ [M+H]+ 604.2.
Step c. To a solution of (3aR,11aS)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-5-(2-((S)-1-oxooctahydro-2H-pyrazino[1,2-a]pyrazin-2-yl)ethyl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (60 mg, 0.084 mmol) and TEA (25 mg, 0.25 mmol) in DCM (2 mL) was added acryloyl chloride (8 mg, 0.084 mmol). The mixture was stirred at 20° C. for 30 min. Upon completion, the reaction mixture was evaporated. The resulting residue was purified by Prep-HPLC to give the title compound (39 mg, 70% yield) as white solid.
m/z ES+ [M+H]+ 658.3; 1H NMR (400 MHz, CDCl3) δ ppm 8.32 (s, 1H), 7.40-7.32 (m, 1H), 7.23 (d, J=8.0 Hz, 1H), 7.19-7.12 (m, 1H), 7.06 (s, 1H), 6.73-6.54 (m, 1H), 6.34 (dd, J=1.2, 16.8 Hz, 1H), 5.80-5.77 (m, 1H), 4.63-4.45 (m, 2H), 4.08-3.89 (m, 1H), 3.78-3.57 (m, 3H), 3.45-3.00 (m, 13H), 2.92-2.69 (m, 4H), 2.50 (s, 3H), 2.25-2.17 (m, 1H).
The Examples in Table 6 were prepared using methods similar to those described in the synthesis of Example 45, using the listed Intermediates in step a.
For Example 49 and Example 50, step a using tert-butyl 1,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridine-5-carboxylate (CAS: 1202800-68-7) formed a mixture of regioisomers which were separated by chiral SFC and used in the subsequent steps.
1H NMR (400 MHz) δ ppm
Step a. To a solution of Intermediate 9a (90 mg, 0.19 mmol) in DCM (2 mL) was added DIPEA (73 mg, 0.56 mmol), HATU (86 mg, 0.23 mmol) and Intermediate A1 (90 mg, 0.38 mmol). The mixture was stirred at 20° C. for 1 h. Upon completion, the reaction mixture was diluted with water (5 mL) and extracted with EtOAc (3×2.5 mL). The combined organic layers were washed with brine (5 mL), dried over Na2SO4 and evaporated. The crude product was purified by Prep-TLC (EtOAc/MeOH=10/1) to give tert-butyl 3-(4-(2-((3aR,11aS)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)acetyl)piperazin-1-yl)azetidine-1-carboxylate (100 mg, 75% yield) as white solid.
m/z ES+ [M+H]+ 704.2; 1H NMR (400 MHz, CDCl3) δ ppm 8.33 (s, 1H), 7.38-7.35 (m, 1H), 7.24-7.22 (m, 1H), 7.17-7.15 (m, 1H), 7.06 (s, 1H), 4.64 (d, J=9.2 Hz, 1H), 4.03-3.90 (m, 3H), 3.84-3.73 (m, 3H), 3.71-3.43 (m, 5H), 3.39 (s, 3H), 3.14-3.04 (m, 1H), 3.01-2.86 (m, 2H), 2.75-2.70 (m, 1H), 2.49 (s, 3H), 2.42-2.15 (m, 5H), 1.43 (s, 9H).
Step b. To a solution of tert-butyl 3-(4-(2-((3aR,11aS)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)acetyl)piperazin-1-yl)azetidine-1-carboxylate (100 mg, 0.14 mmol) in DCM (1 mL) was added TFA (1.54 g, 13.5 mmol). The mixture was stirred at 20° C. for 30 min. Upon completion, the reaction mixture was evaporated to give (3aR,11aS)-5-(2-(4-(azetidin-3-yl)piperazin-1-yl)-2-oxoethyl)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione as a TFA salt (100 mg, crude) as yellow oil.
m/z ES+ [M+H]+ 604.3.
Step c. To a solution of (3aR,11aS)-5-(2-(4-(azetidin-3-yl)piperazin-1-yl)-2-oxoethyl)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (100 mg, 0.14 mmol) in DCM (2 mL) was added TEA (42 mg, 0.42 mmol) and acryloyl chloride (13 mg, 0.14 mmol). The mixture was stirred at 20° C. for 30 min. Upon completion, the reaction mixture was evaporated. The crude product was purified by Prep-HPLC to give the title compound (41 mg, 43% yield) as a white solid.
m/z ES+ [M+H]+ 658.3; 1H NMR (400 MHz, CDCl3) δ ppm 8.32 (s, 1H), 7.39-7.36 (m, 1H), 7.25-7.23 (m, 1H), 7.17 (t, J=9.2 Hz, 1H), 7.06 (s, 1H), 6.38-6.28 (m, 1H), 6.22-6.12 (m, 1H), 5.70-5.67 (m, 1H), 4.63 (d, J=8.8 Hz, 1H), 4.30-4.22 (m, 1H), 4.19-4.08 (m, 2H), 4.06-3.93 (m, 2H), 3.81-3.77 (m, 2H), 3.66-3.46 (m, 4H), 3.39 (s, 3H), 3.32-3.22 (m, 1H), 2.99-2.87 (m, 2H), 2.78-2.74 (m, 1H), 2.59-2.29 (m, 7H), 2.24-2.19 (m, 1H).
The Example in Table 7 was prepared using methods similar to those described in the synthesis of Example 51, using the listed Intermediates in step a.
1H NMR (400 MHz) δ ppm
Step a. A mixture of Intermediate 5a (200 mg, 0.47 mmol), tert-butyl 4-(4-bromopyridin-2-yl)piperazine-1-carboxylate (CAS: 1197294-80-6; 243 mg, 0.71 mmol), RuPhos (44 mg, 0.095 mmol), Cs2CO3 (462 mg, 1.42 mmol) and RuPhos-Pd-G2 (37 mg, 0.047 mmol) in 1,4-dioxane (10 mL) was stirred at 100° C. for 20 h under an N2 atmosphere. Upon completion, the reaction mixture was diluted with water (50 mL) and extracted with EtOAc (3×20 mL). The combined organic layers were washed with brine (50 mL), dried over Na2SO4 and evaporated. The crude product was purified by reverse phase flash chromatography (water (0.1 FA)/MeCN) to give tert-butyl 4-(4-((3aR,11aS)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)pyridin-2-yl)piperazine-1-carboxylate (80 mg, 24% yield) as a yellow solid.
m/z ES+ [M+H]+ 684.4; 1H NMR (400 MHz, CDCl3) δ ppm 8.32 (s, 1H), 7.89 (d, J=6.0 Hz, 1H), 7.64-7.54 (m, 1H), 7.39-7.31 (m, 2H), 7.08 (s, 1H), 5.79-5.71 (m, 1H), 5.58 (s, 1H), 4.82 (d, J=9.2 Hz, 1H), 4.52-4.40 (m, 1H), 3.55-3.51 (m, 4H), 3.48-3.39 (m, 4H), 3.24 (s, 1H), 3.19 (s, 3H), 3.15-3.07 (m, 1H), 2.96-2.86 (m, 1H), 2.50 (s, 3H), 2.40-2.29 (m, 1H), 1.48 (s, 9H).
Step b. To a mixture of tert-butyl 4-(4-((3aR,11aS)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)pyridin-2-yl)piperazine-1-carboxylate (99 mg, 0.14 mmol) in DCM (5 mL) was added TFA (3.81 g, 33.4 mmol). The reaction mixture was stirred at 25° C. for 1 h. Upon completion, the reaction mixture was evaporated to give (3aR,11aS)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-5-(2-(piperazin-1-yl)pyridin-4-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione as a TFA salt (100 mg, 99% yield) as a yellow oil.
m/z ES+ [M+H]+ 584.2.
Step c. To a mixture of (3aR,11aS)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-5-(2-(piperazin-1-yl)pyridin-4-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (100 mg, 0.14 mmol) and DIPEA (37 mg, 0.29 mmol) in DCM (4 mL) was added acryloyl chloride (14 mg, 0.16 mmol). The reaction mixture was stirred at 25° C. for 30 min. Upon completion, the reaction mixture was quenched with water (50 mL) and extracted with DCM (3×20 mL). The combined organic layers were washed with brine (50 mL), dried over Na2SO4 and evaporated. The crude product was purified by Prep-HPLC to give the title compound (60 mg, 63% yield) as white solid.
m/z ES+ [M+H]+ 638.3; 1H NMR (400 MHz, CDCl3) δ ppm 8.32 (s, 1H), 7.88 (d, J=6.0 Hz, 1H), 7.69-7.59 (m, 1H), 7.45-7.33 (m, 2H), 7.09 (s, 1H), 6.63-6.53 (m, 1H), 6.39-6.30 (m, 1H), 5.94-5.50 (m, 3H), 4.82 (d, J=9.6 Hz, 1H), 4.56-4.46 (m, 1H), 3.90-3.68 (m, 6H), 3.54-3.36 (m, 2H), 3.34-3.25 (m, 1H), 3.20 (s, 3H), 3.13-3.03 (m, 1H), 2.98-2.89 (m, 1H), 2.51 (s, 3H), 2.42-2.31 (m, 1H).
The Examples in Table 8 were prepared using methods similar to those described in the synthesis of Example 53, using the listed Intermediates in step a.
1H NMR (400 MHz) δ ppm
Step a. This step was conducted in a similar manner to Example 1, step a, using Intermediate 7a and Intermediate A19a.
Step b. A mixture of benzyl (3R/S,9aR/S)-8-(2-((3aR,11aS)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)ethyl)-3-methyl-4-oxooctahydro-2H-pyrazino[1,2-a]pyrazine-2-carboxylate (50 mg, 0.07 mmol) and 10% Pd/C (5 mg) in MeOH (1.5 mL) was stirred at 30° C. for 1 h under a H2 atmosphere (15 psi). Upon completion, the reaction mixture was filtered and evaporated to give (3aR,11aS)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-5-(2-((7R/S,9aS/R)-7-methyl-6-oxooctahydro-2H-pyrazino[1,2-a]pyrazin-2-yl)ethyl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (40 mg, crude) as a yellow solid.
m/z ES+ [M+H]+ 618.3.
Step c. This step was conducted in a similar manner to Example 1, step c.
m/z ES+ [M+H]+ 672.3; 1H NMR (400 MHz, CDCl3) δ ppm 8.32 (s, 1H), 7.40-7.31 (m, 1H), 7.24 (d, J=7.6 Hz, 1H), 7.15 (t, J=9.2 Hz, 1H), 7.06 (s, 1H), 6.66-6.32 (m, 2H), 5.79 (d, J=10.4 Hz, 1H), 5.21-4.68 (m, 1H), 4.67-4.39 (m, 3H), 3.57 (d, J=9.2 Hz, 1H), 3.53-3.43 (m, 1H), 3.37 (s, 3H), 3.32-3.11 (m, 2H), 3.00-2.81 (m, 4H), 2.81-2.63 (m, 3H), 2.50 (s, 3H), 2.48-2.38 (m, 2H), 2.26-2.16 (m, 1H), 2.12-1.95 (m, 1H), 1.80-1.72 (m, 1H), 1.52 (d, J=6.4 Hz, 3H).
The title compound was prepared in a similar manner to Example 58, using Intermediate A19b in step a.
m/z ES+ [M+H]+ 672.3; 1H NMR (400 MHz, CDCl3) δ ppm 8.32 (s, 1H), 7.38-7.31 (m, 1H), 7.23 (d, J=7.6 Hz, 1H), 7.15 (t, J=9.2 Hz, 1H), 7.06 (s, 1H), 6.54-6.35 (m, 2H), 5.79 (d, J=11.2 Hz, 1H), 4.80-4.65 (m, 1H), 4.64-4.42 (m, 3H), 3.58-3.53 (m, 1H), 3.52-3.50 (m, 1H), 3.36 (s, 3H), 3.28-3.21 (m, 2H), 2.97-2.82 (m, 4H), 2.80-2.68 (m, 3H), 2.50 (s, 3H), 2.48-2.38 (m, 2H), 2.25-2.17 (m, 1H), 2.08-1.98 (m, 1H), 1.87-1.72 (m, 1H), 1.57-1.47 (m, 3H).
The title compound was prepared in a similar manner to Example 58, using Intermediate A19c in step a.
m/z ES+ [M+H]+ 672.1; 1H NMR (400 MHz, CDCl3) δ ppm 8.33 (s, 1H), 7.39-7.30 (m, 1H), 7.23 (d, J=8.0 Hz, 1H), 7.14 (t, J=9.2 Hz, 1H), 7.06 (s, 1H), 6.55-6.45 (m, 1H), 6.45-6.33 (m, 1H), 5.79 (d, J=10.0 Hz, 1H), 5.41-4.61 (m, 1H), 4.61-4.34 (m, 3H), 4.04-3.58 (m, 1H), 3.58-3.44 (m, 2H), 3.37 (s, 3H), 3.32-3.24 (m, 1H), 3.20-3.08 (m, 1H), 3.00-2.86 (m, 2H), 2.85-2.79 (m, 1H), 2.77-2.63 (m, 3H), 2.50 (s, 3H), 2.40-2.30 (m, 2H), 2.20 (dd, J=16.8, 11.6 Hz, 1H), 2.10-1.93 (m, 2H), 1.51 (d, J=6.8 Hz, 3H).
The title compound was prepared in a similar manner to Example 58, using Intermediate A19d in step a.
m/z ES+ [M+H]+ 672.3; 1H NMR (400 MHz, CDCl3) δ ppm 8.32 (s, 1H), 7.37-7.32 (m, 1H), 7.23 (d, J=8.0 Hz, 1H), 7.11 (t, J=9.2 Hz, 1H), 7.06 (s, 1H), 6.53-6.37 (m, 2H), 5.80-5.78 (m, 2H), 4.61-4.50 (m, 3H), 3.55-3.48 (m, 2H), 3.35 (s, 3H), 3.25-3.17 (m, 2H), 2.90-2.68 (m, 6H), 2.50 (s, 3H), 2.40-2.30 (m, 2H), 2.24-2.17 (m, 1H), 2.10-1.93 (m, 2H), 1.51-1.28 (m, 4H).
The title compounds were prepared in a similar manner to Example 58, using Intermediate A22 in step a, and an additional chiral SFC step was conducted after step a.
Example 62a:
m/z ES+ [M+H]+ 658.3; 1H NMR (400 MHz, CDCl3) δ ppm 8.32 (s, 1H), 7.40-7.30 (m, 1H), 7.23 (d, J=8.0 Hz, 1H), 7.20-7.10 (m, 1H), 7.06 (s, 1H), 6.60-6.43 (m, 1H), 6.42-6.31 (m, 1H), 5.83-5.75 (m, 1H), 4.60 (d, J=9.2 Hz, 1H), 4.51 (d, J=11.6 Hz, 1H), 4.43-3.89 (m, 3H), 3.65-3.43 (d, J=9.2 Hz, 2H), 3.36 (s, 3H), 3.32-3.13 (m, 3H), 3.02-2.63 (m, 6H), 2.50 (s, 3H), 2.46-2.31 (m, 2H), 2.26-2.14 (m, 1H), 2.10-1.81 (m, 2H).
Example 62b:
m/z ES+ [M+H]+ 658.3; 1H NMR (400 MHz, CDCl3) δ ppm 8.32 (s, 1H), 7.39-7.31 (m, 1H), 7.25-7.22 (m, 1H), 7.15 (t, J=9.2 Hz, 1H), 7.06 (s, 1H), 6.61-6.43 (m, 1H), 6.41-6.33 (m, 1H), 5.81-5.76 (m, 1H), 4.60 (d, J=9.2 Hz, 1H), 4.57-4.46 (m, 1H), 4.39-4.27 (m, 1H), 4.12-4.05 (m, 1H), 3.58-3.50 (m, 2H), 3.36 (s, 3H), 3.33-3.11 (m, 3H), 2.99-2.65 (m, 6H), 2.50 (s, 3H), 2.45-2.35 (m, 2H), 2.26-2.14 (m, 1H), 2.08-2.01 (m, 1H), 1.94-1.78 (m, 1H), 1.72-1.60 (m, 1H).
The title compounds were prepared in a similar manner to Example 58, using Intermediate A24a in step a, and an additional chiral SFC step was conducted after step a.
Example 63a:
m/z ES+ [M+H]+ 658.4; 1H NMR (400 MHz, CDCl3) δ ppm 8.33 (s, 1H), 7.38-7.30 (m, 1H), 7.22 (d, J=8.0 Hz, 1H), 7.15 (t, J=9.2 Hz, 1H), 7.06 (s, 1H), 6.66-6.50 (m, 1H), 6.38-6.21 (m, 1H), 5.71 (d, J=8.8 Hz, 1H), 4.60 (d, J=9.2 Hz, 1H), 4.56-4.40 (m, 1H), 3.83-3.67 (m, 1H), 3.62-3.53 (m, 1H), 3.35 (s, 3H), 3.32-3.22 (m, 1H), 3.22-3.04 (m, 2H), 3.03-2.56 (m, 7H), 2.50 (s, 3H), 2.48-2.25 (m, 3H), 2.25-2.13 (m, 3H), 2.13-2.00 (m, 1H), 1.94-1.71 (m, 1H), 1.09 (d, J=5.6 Hz, 3H).
Example 63b:
m/z ES+ [M+H]+ 658.3; 1H NMR (400 MHz, CDCl3) δ ppm 8.31 (s, 1H), 7.38-7.29 (m, 1H), 7.22 (d, J=8.0 Hz, 1H), 7.14 (t, J=9.2 Hz, 1H), 7.05 (s, 1H), 6.55 (dd, J=16.8, 10.4 Hz, 1H), 6.28 (dd, J=16.8, 1.6 Hz, 1H), 5.70 (d, J=10.4 Hz, 1H), 4.60 (d, J=9.2 Hz, 1H), 4.55-4.32 (m, 1H), 3.85-3.64 (m, 1H), 3.60-3.50 (m, 1H), 3.36 (s, 3H), 3.31-3.18 (m, 2H), 3.13-3.05 (m, 1H), 3.00-2.79 (m, 4H), 2.75-2.65 (m, 2H), 2.51-2.48 (m, 4H), 2.45-2.30 (m, 3H), 2.25-2.15 (m, 3H), 2.12-1.98 (m, 1H), 1.97-1.83 (m, 1H), 1.09 (s, 3H).
The title compounds were prepared in a similar manner to Example 58, using Intermediate A24b in step a, and an additional chiral SFC step was conducted after step a.
Example 64a:
m/z ES+ [M+H]+ 658.4; 1H NMR (400 MHz, CDCl3) δ ppm 8.32 (s, 1H), 7.37-7.29 (m, 1H), 7.22 (d, J=7.6 Hz, 1H), 7.18-7.10 (m, 1H), 7.05 (s, 1H), 6.67-6.45 (m, 1H), 6.38-6.22 (m, 1H), 5.70 (dd, J=10.4, 1.6 Hz, 1H), 4.60 (d, J=9.2 Hz, 1H), 4.50-4.38 (m, 1H), 3.83-3.68 (m, 1H), 3.64-3.40 (m, 2H), 3.35 (s, 3H), 3.30-3.13 (m, 2H), 3.08-2.98 (m, 1H), 2.96-2.86 (m, 2H), 2.83-2.54 (m, 6H), 2.49 (s, 3H), 2.46-2.13 (m, 5H), 1.89-1.70 (m, 1H), 0.99 (d, J=6.4 Hz, 3H).
Example 64b:
m/z ES+ [M+H]+ 658.4; 1H NMR (400 MHz, CDCl3) δ ppm 8.33 (s, 1H), 7.38-7.31 (m, 1H), 7.23 (d, J=7.6 Hz, 1H), 7.20-7.12 (m, 1H), 7.07 (s, 1H), 6.70-6.45 (m, 1H), 6.31 (d, J=16.8 Hz, 1H), 5.71 (d, J=10.4 Hz, 1H), 4.62 (d, J=9.2 Hz, 1H), 4.46 (d, J=11.2 Hz, 1H), 3.73 (d, J=12.4 Hz, 1H), 3.63-3.51 (m, 1H), 3.37 (s, 3H), 3.31-3.13 (m, 2H), 3.10-2.82 (m, 4H), 2.81-2.60 (m, 5H), 2.58-2.50 (m, 4H), 2.48-2.28 (m, 3H), 2.21 (dd, J=16.8, 11.2 Hz, 2H), 1.88-1.77 (m, 1H), 0.99 (d, J=6.4 Hz, 3H).
Step a. This step was conducted in a similar manner to Example 1, step a, using Intermediate 7b and methanamine (2 M in THF).
Step b. To a solution of (3aR,11aS)-6-chloro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-5-(2-(methylamino)ethyl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (60 mg, 0.093 mmol) and 1-(tert-butoxycarbonyl)azetidine-3-carboxylic acid (24 mg, 0.12 mmol) in DCM (1.2 mL) was added DIPEA (24 mg, 0.19 mmol) and HATU (43 mg, 0.11 mmol). The mixture was stirred at 20° C. for 30 min. Upon completion, the reaction mixture was quenched with H2O (5 mL) and extracted with EtOAc (3×5 mL). The combined organic layers were washed with brine (15 mL), dried over Na2SO4 and evaporated. The residue was purified by Prep-TLC (EtOAc) to give tert-butyl 3-((2-((3aR,11aS)-6-chloro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)ethyl)(methyl)carbamoyl)azetidine-1-carboxylate (50 mg, 79% yield) as a yellow solid.
m/z ES+ [M+H]+ 679.1.
Steps c-d. These 2 steps were conducted in a similar manner to Example 27, steps b-c, using Intermediate E1 in step d.
m/z ES+ [M+H]+ 690.1; 1H NMR (400 MHz, CDCl3) δ ppm 8.33 (s, 1H), 7.50-7.41 (m, 1H), 7.37-7.29 (m, 2H), 7.05 (s, 1H), 5.91 (s, 1H), 5.73 (s, 1H), 4.67-4.51 (m, 2H), 4.45-4.28 (m, 2H), 4.19-4.03 (m, 1H), 3.78-3.50 (m, 4H), 3.42-3.32 (m, 4H), 3.25-3.08 (m, 3H), 3.01-2.87 (m, 5H), 2.80-2.72 (m, 1H), 2.62-2.54 (m, 6H), 2.49-2.46 (m, 3H), 2.24-2.17 (m, 1H).
Step a. This step was conducted in a similar manner to Example 1, step a, using Intermediate 7b and tert-butyl (3R,4S)-4-amino-3-fluoropiperidine-1-carboxylate (CAS: 907544-17-6).
Step b. To a solution of tert-butyl (3R,4S)-4-((2-((3aR,11aS)-6-chloro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)ethyl)amino)-3-fluoropiperidine-1-carboxylate (120 mg, 0.15 mmol) in THF (1.5 mL) and H2O (1.5 mL) was added NaHCO3 (26 mg, 0.31 mmol) and CbzCl (34 mg, 0.2 mmol). The mixture was stirred at 25° C. for 30 min. Upon completion, the reaction mixture was quenched with H2O (5 mL) and extracted with EtOAc (3×5 mL). The combined organic layers were washed with brine (12 mL), dried over Na2SO4 and evaporated. The residue was purified by Prep-TLC (PE/EtOAc=1/1) to give tert-butyl (3R,4S)-4-(((benzyloxy)carbonyl)(2-((3aR,11aS)-6-chloro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)ethyl)amino)-3-fluoropiperidine-1-carboxylate (110 mg, 86% yield) as an off-white solid.
m/z ES+ [M+H]+ 817.3; 1H NMR (400 MHz, CDCl3) δ ppm 8.31 (s, 1H), 7.43 (d, J=8.8 Hz, 1H), 7.40-7.27 (m, 6H), 7.26 (s, 1H), 7.04 (s, 1H), 5.15-5.06 (m, 2H), 4.68-4.58 (m, 1H), 4.53 (d, J=9.2 Hz, 1H), 4.40-4.31 (m, 1H), 4.27-4.17 (m, 1H), 3.50-3.22 (m, 4H), 3.16 (s, 3H), 3.07-2.87 (m, 3H), 2.85-2.69 (m, 3H), 2.53-2.42 (m, 4H), 2.04-2.02 (m, 1H), 1.55-1.47 (m, 2H), 1.46 (s, 9H).
Step c. This step was conducted in a similar manner to Example 1, step b.
Step d. To a solution of benzyl (2-((3aR,11aS)-6-chloro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)ethyl)((3R,4S)-3-fluoropiperidin-4-yl)carbamate (96 mg, 0.13 mmol) and acrylic acid (13 mg, 0.17 mmol) in DCM (2 mL) was added DIPEA (35 mg, 0.27 mmol) and HATU (61 mg, 0.16 mmol). The mixture was stirred at 25° C. for 1 h. Upon completion, the reaction mixture was quenched with H2O (5 mL) and extracted with EtOAc (3×5 mL). The combined organic layers were washed with brine (10 mL), dried over Na2SO4 and evaporated. The residue was purified by Prep-TLC (EtOAc) to give benzyl ((3R,4S)-1-acryloyl-3-fluoropiperidin-4-yl)(2-((3aR,11aS)-6-chloro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)ethyl)carbamate (45 mg, 42% yield) as a white solid.
m/z ES+ [M+H]+ 771.2.
Step e. A solution of benzyl ((3R,4S)-1-acryloyl-3-fluoropiperidin-4-yl)(2-((3aR,11aS)-6-chloro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)ethyl)carbamate (40.0 mg, 0.05 mmol) in TFA (1 mL) was stirred at 45° C. for 12 h. Upon completion, the reaction mixture was evaporated. The residue was purified by Prep-HPLC to give the title compound (11 mg, 33% yield) as a white solid.
m/z ES+ [M+H]+ 637.2; 1H NMR (400 MHz, CDCl3) δ ppm 8.33 (s, 1H), 7.46 (d, J=6.4 Hz, 1H), 7.35-7.28 (m, 2H), 7.06 (s, 1H), 6.65-6.50 (m, 1H), 6.35-6.23 (m, 1H), 5.71 (dd, J=10.4, 1.6 Hz, 1H), 4.98-4.76 (m, 1H), 4.71-4.56 (m, 2H), 4.37-3.95 (m, 1H), 3.59 (d, J=10.0 Hz, 1H), 3.39 (s, 3H), 3.33-3.11 (m, 3H), 3.02-2.81 (m, 4H), 2.80-2.70 (m, 2H), 2.65-2.56 (m, 1H), 2.48 (s, 3H), 2.28-2.20 (m, 1H), 1.83-1.77 (m, 2H), 1.72-1.65 (m, 1H).
Step a. This step was conducted as described in Example 66, step a.
Step b. To a solution of tert-butyl (3R,4S)-4-((2-((3aR,11aS)-6-chloro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)ethyl)amino)-3-fluoropiperidine-1-carboxylate (55 mg, 0.08 mmol) and paraformaldehyde (24 mg, 0.81 mmol) in MeOH (2 mL) was added NaBH3CN (15 mg, 0.24 mmol) and acetic acid (10 mg, 0.16 mmol). The mixture was stirred at 40° C. for 12 h. Upon completion, the reaction mixture was diluted with sat. aq. NaHCO3 (5 mL), and then extracted with EtOAc (3×5 mL). The combined organic layers were washed with brine (15 mL), dried over Na2SO4 and evaporated. The crude product was purified by Prep-TLC (PE/EtOAc=1/1) to give tert-butyl (3R,4S)-4-((2-((3aR,11aS)-6-chloro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)ethyl)(methyl)amino)-3-fluoropiperidine-1-carboxylate (45 mg, 78% yield) as a white solid.
m/z ES+ [M+H]+ 697.3; 1H NMR (400 MHz, CDCl3) δ ppm 8.33 (s, 1H), 7.45 (d, J=7.2 Hz, 1H), 7.36-7.29 (m, 2H), 7.05 (s, 1H), 4.94-4.68 (m, 1H), 4.65-4.58 (m, 1H), 4.47-4.26 (m, 2H), 3.59-3.56 (m, 1H), 3.38 (s, 3H), 3.23-3.07 (m, 2H), 3.05-2.86 (m, 3H), 2.82-2.54 (m, 5H), 2.48 (s, 3H), 2.35-2.25 (m, 2H), 2.25-2.20 (m, 1H), 1.97-1.86 (m, 1H), 1.64-1.54 (m, 2H), 1.46 (s, 9H).
Steps c-d. These 2 steps were conducted in a similar manner to Example 1, steps b-c.
m/z ES+ [M+H]+ 651.3; 1H NMR (400 MHz, CDCl3) δ ppm 8.33 (s, 1H), 7.47-7.43 (m, 1H), 7.35-7.28 (m, 2H), 7.05 (s, 1H), 6.65-6.48 (m, 1H), 6.35-6.20 (m, 1H), 5.71 (d, J=10.4 Hz, 1H), 5.11-4.91 (m, 1H), 4.90-4.77 (m, 1H), 4.61 (d, J=9.2 Hz, 1H), 4.29-4.10 (m, 1H), 3.57 (d, J=10.0 Hz, 1H), 3.38 (s, 3H), 3.31-3.05 (m, 3H), 3.04-2.93 (m, 2H), 2.86-2.53 (m, 5H), 2.49 (s, 3H), 2.48-2.27 (m, 3H), 2.21 (dd, J=16.8, 11.2 Hz, 1H), 2.02-1.89 (m, 1H), 1.75-1.65 (m, 1H).
Steps a-b. These 2 steps were conducted in a similar manner to Example 66, steps c-d, using Intermediate 12a.
m/z ES+ [M+H]+ 635.3; 1H NMR (400 MHz, CDCl3) δ ppm 8.34 (s, 1H), 7.50-7.45 (m, 1H), 7.36-7.29 (m, 2H), 7.06 (s, 1H), 6.38-6.28 (m, 1H), 6.24-6.13 (m, 1H), 5.67 (d, J=10.4 Hz, 1H), 4.66-4.58 (m, 1H), 4.30-4.18 (m, 1H), 4.15-3.99 (m, 2H), 3.96-3.86 (m, 1H), 3.74-3.56 (m, 2H), 3.53-3.41 (m, 1H), 3.39 (s, 3H), 3.32-3.14 (m, 2H), 3.09-3.01 (m, 1H), 3.00-2.86 (m, 2H), 2.80-2.71 (m, 1H), 2.49 (s, 3H), 2.46-2.38 (m, 1H), 2.34-2.28 (m, 1H), 2.26 (s, 3H), 2.24-2.18 (m, 1H).
The title compound was prepared in a similar manner to Example 68, using Intermediate 12b in step a.
m/z ES+ [M+H]+ 635.3; 1H NMR (400 MHz, CDCl3) δ ppm 8.34 (s, 1H), 7.47 (dd, J=7.2, 2.4 Hz, 1H), 7.38-7.29 (m, 2H), 7.06 (s, 1H), 6.39-6.28 (m, 1H), 6.24-6.12 (m, 1H), 5.68 (d, J=10.4 Hz, 1H), 4.71-4.57 (m, 1H), 4.30-4.15 (m, 1H), 4.16-4.02 (m, 2H), 4.00-3.85 (m, 1H), 3.83-3.58 (m, 2H), 3.55-3.25 (m, 5H), 3.16-2.89 (m, 4H), 2.76 (dd, J=16.8, 7.6 Hz, 1H), 2.49 (s, 3H), 2.46-2.16 (m, 6H).
Step a. To a solution of Intermediate 13 (0.17 g, 0.27 mmol) in THF (3 mL) and water (1 mL) was added CbzCl (69 mg, 0.40 mmol) and Na2CO3 (85 mg, 0.80 mmol). The mixture was stirred at 25° C. for 1 h. Upon completion, the reaction mixture was quenched with sat. aq. NH4Cl (20 mL), diluted with water (30 mL) and extracted with EtOAc (3×30 mL). The combined organic layers were washed with brine (20 mL), dried over Na2SO4 and evaporated. Purification by column chromatography (PE/EtOAc=1/1) afforded tert-butyl 3-(((benzyloxy)carbonyl)(3-((3aR,11aS)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)propyl)amino)azetidine-1-carboxylate (180 mg, 87% yield) as a yellow solid.
m/z ES+ [M+H]+ 769.1; 1H NMR (400 MHz, CDCl3) δ ppm 8.33 (s, 1H), 7.38-7.30 (m, 6H), 7.20 (d, J=8.4 Hz, 1H), 7.13 (t, J=9.2 Hz, 1H), 7.05 (s, 1H), 5.12 (s, 2H), 4.61 (d, J=8.4 Hz, 2H), 4.26-4.02 (m, 3H), 3.99-3.85 (m, 2H), 3.51-3.40 (m, 1H), 3.32-3.25 (m, 5H), 3.10-2.96 (m, 2H), 2.90-2.75 (m, 2H), 2.70-2.57 (m, 1H), 2.49 (s, 3H), 2.20-2.13 (m, 1H), 1.56-1.51 (m, 1H), 1.44 (s, 9H).
Steps b-c. These 2 steps were conducted in a similar manner to Example 67, steps c-d.
Step d. To a solution of benzyl (1-acryloylazetidin-3-yl)(3-((3aR,11aS)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)propyl)carbamate (150 mg, 0.21 mmol) in DCM (3 mL) was added BCl3 (1 M in DCM, 2.08 mL) and the mixture was stirred at 0° C. for 30 min. Upon completion, the reaction mixture was quenched with sat. aq. NaHCO3 (30 mL), diluted with water (10 mL) and extracted with DCM (3×30 mL). The combined organic layers were washed with sat. aq. NaHCO3 (3×20 mL), dried over Na2SO4 and evaporated. The crude product was purified by Prep-HPLC to give the title compound (55 mg, 45% yield) as a white solid.
m/z ES+ [M+H]+ 589.1; 1H NMR (400 MHz, CDCl3) δ ppm 8.32 (s, 1H), 7.37-7.30 (m, 1H), 7.22 (d, J=8.0 Hz, 1H), 7.14 (t, J=9.2 Hz, 1H), 7.05 (s, 1H), 6.35-6.28 (m, 1H), 6.23-6.14 (m, 1H), 5.65 (d, J=10.4 Hz, 1H), 4.61 (d, J=9.2 Hz, 1H), 4.38-4.31 (m, 1H), 4.26-4.18 (m, 1H), 3.91-3.84 (m, 1H), 3.74-3.70 (m, 1H), 3.65-3.62 (m, 1H), 3.56-3.52 (m, 1H), 3.35 (s, 3H), 3.23-3.08 (m, 2H), 2.95-2.82 (m, 2H), 2.74-2.69 (m, 1H), 2.67-2.55 (m, 2H), 2.49 (s, 3H), 2.24-2.16 (m, 1H), 1.54-1.47 (m, 3H).
Step a. To a solution of Intermediate 13 (100 mg, 0.16 mmol) in MeOH (2 mL) was added formaldehyde (37 wt. % in H2O, 2.18 g, 26.9 mmol), acetic acid (47 mg, 0.79 mmol) and NaBH3CN (20 mg, 0.32 mmol). The mixture was stirred at 25° C. for 16 h. Upon completion, the reaction mixture was diluted with water (10 mL) and EtOAc (5 mL). The layers were separated and the aqueous layer was further extracted with EtOAc (2×5 mL), dried over Na2SO4 and evaporated to give tert-butyl 3-((3-((3aR,11aS)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)propyl)(methyl)amino)azetidine-1-carboxylate (100 mg, crude) as yellow oil.
m/z ES+ [M+H]+ 649.2.
Steps b-c. These 2 steps were conducted in a similar manner to Example 67, steps c-d.
m/z ES+ [M+H]+ 603.3; 1H NMR (400 MHz, CDCl3) δ ppm 8.33 (s, 1H), 7.38-7.31 (m, 1H), 7.22 (d, J=8.0 Hz, 1H), 7.19-7.11 (m, 1H), 7.06 (s, 1H), 6.37-6.28 (m, 1H), 6.25-6.14 (m, 1H), 5.68 (m, 1H), 4.62 (d, J=9.2 Hz, 1H), 4.34-3.84 (m, 4H), 3.57-3.54 (m, 1H), 3.43-3.30 (m, 4H), 3.05 (m, 2H), 2.97-2.68 (m, 4H), 2.50 (s, 4H), 2.34-2.12 (m, 4H), 1.68-1.51 (m, 2H).
Step a. This step was conducted in a similar manner to Example 1, step a, using Intermediate 7i and Intermediate A25.
Step b. To a mixture of (3aR,11aS)-5-(2-(4-(1-benzhydryl-3-methylazetidin-3-yl)piperazin-1-yl)ethyl)-6,10-dimethyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (100 mg, 0.13 mmol) in 2-propanol (10 mL) was added 10% Pd/C (20 mg), 10% Pd(OH)2/C (20 mg) and 4 M HCl in dioxane (33 uL). The reaction mixture was stirred at 40° C. for 28 hr under a H2 atmosphere (15 psi). Upon completion, the reaction mixture was filtered through a pad of Celite, washing with MeOH (2×3 mL). The filtrate was evaporated to give (3aR,11aS)-6,10-dimethyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-5-(2-(4-(3-methylazetidin-3-yl)piperazin-1-yl)ethyl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (60 mg, 76% yield) as a yellow oil.
m/z ES+ [M+H]+ 600.3.
Step c. This step was conducted in a similar manner to Example 1, step c.
m/z ES+ [M+H]+ 654.1; 1H NMR (400 MHz, CDCl3) δ ppm 8.33 (s, 1H), 7.27-7.19 (m, 3H), 7.05 (s, 1H), 6.38-6.29 (m, 1H), 6.24-6.12 (m, 1H), 5.70-5.63 (m, 1H), 4.69 (d, J=9.2 Hz, 1H), 4.02 (d, J=8.0 Hz, 1H), 3.88 (d, J=9.6 Hz, 1H), 3.79 (d, J=7.6 Hz, 1H), 3.72-3.58 (m, 2H), 3.38 (s, 3H), 3.27-2.87 (m, 4H), 2.86-2.67 (m, 3H), 2.60-2.50 (m, 2H), 2.49 (s, 3H), 2.48-2.40 (m, 4H), 2.39 (s, 3H), 2.38-2.26 (m, 2H), 2.24-2.14 (m, 1H), 1.32 (s, 3H).
The title compounds were prepared in a similar manner to Example 39, using Intermediate 5b and Intermediate B26 in step a, and 2-chloroacetyl chloride in step c. An additional chiral SFC step was conducted after step a.
Example 73a:
m/z ES+ [M+H]+ 664.2; 1H NMR (400 MHz, CDCl3) δ ppm 8.31 (s, 1H), 7.47 (dd, J=7.2, 2.4 Hz, 1H), 7.35-7.29 (m, 2H), 7.06 (s, 1H), 6.90 (s, 1H), 5.44-5.22 (m, 1H), 5.00-4.47 (m, 3H), 4.41-4.20 (m, 2H), 4.19-4.10 (m, 3H), 3.96-3.83 (m, 1H), 3.67-3.61 (m, 1H), 3.33 (s, 3H), 3.10-3.03 (m, 1H), 3.02-2.93 (m, 1H), 2.69 (dd, J=16.8, 8.0 Hz, 1H), 2.49 (s, 3H), 2.20 (dd, J=16.8, 11.6 Hz, 1H), 1.36-1.20 (m, 3H).
Example 73b:
m/z ES+ [M+H]+ 664.1; 1H NMR (400 MHz, CDCl3) δ ppm 8.31 (s, 1H), 7.47 (dd, J=6.8, 2.8 Hz, 1H), 7.38-7.29 (m, 2H), 7.06 (s, 1H), 6.91 (s, 1H), 5.51-5.24 (m, 1H), 5.11-4.57 (m, 3H), 4.53-4.24 (m, 2H), 4.22-4.13 (m, 3H), 3.95-3.80 (m, 1H), 3.65-3.60 (m, 1H), 3.33 (s, 3H), 3.12-2.90 (m, 2H), 2.70 (dd, J=16.8, 8.4 Hz, 1H), 2.49 (s, 3H), 2.22-2.15 (m, 1H), 1.37-1.19 (m, 3H).
Step a. This step was conducted in a similar manner to Example 39, step a, using Intermediate 5b and Intermediate B19.
Step b. A solution of tert-butyl 2-(((3aR,11aS)-6-chloro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)methyl)-5,6-dihydroimidazo[1,2-a]pyrazine-7(8H)-carboxylate (280 mg, 0.42 mmol) and NBS (70 mg, 0.39 mmol) in MeCN (20 mL) was stirred at 20° C. for 1 h. Upon completion, the mixture was evaporated and the residue was purified by reverse phase flash chromatography (water (0.1% FA)/MeCN) to give tert-butyl 3-bromo-2-(((3aR,11aS)-6-chloro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)methyl)-5,6-dihydroimidazo[1,2-a]pyrazine-7(8H)-carboxylate (160 mg, 51% yield) as a white solid.
m/z ES+ [M+H]+ 754.1.
Step c. A mixture of methylboronic acid (238 mg, 3.98 mmol), tert-butyl 3-bromo-2-(((3aR,11aS)-6-chloro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)methyl)-5,6-dihydroimidazo[1,2-a]pyrazine-7(8H)-carboxylate (150 mg, 0.20 mmol), XPhos-Pd-G2 (31 mg, 0.40 mmol) and Cs2CO3 (194 mg, 0.60 mmol) in toluene (10 mL) was degassed and purged with N2 3 times. The reaction mixture was stirred at 100° C. for 3 h. Upon completion, the mixture was filtered and the filtrate was evaporated. The residue was purified by reverse phase flash chromatography (water (0.1% FA)/MeCN) to give tert-butyl 2-(((3aR,11aS)-6,10-dimethyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)methyl)-3-methyl-5,6-dihydroimidazo[1,2-a]pyrazine-7(8H)-carboxylate (120 mg, 90% yield) as a yellow solid.
m/z ES+ [M+H]+ 668.3.
Steps d-e. These 2 steps were conducted in a similar manner to Example 39, steps b-c.
m/z ES+ [M+H]+ 622.2; 1H NMR (400 MHz, CDCl3) δ ppm 8.36 (s, 1H), 7.34-7.30 (m, 1H), 7.27-7.24 (m, 2H), 7.09 (s, 1H), 6.75-6.60 (m, 1H), 6.43 (d, J=16.4 Hz, 1H), 5.86 (d, J=11.2 Hz, 1H), 4.86 (s, 2H), 4.81-4.74 (m, 1H), 4.65-4.47 (m, 1H), 4.24-4.01 (m, 3H), 3.91-3.83 (m, 2H), 3.65-3.55 (m, 1H), 3.42 (s, 3H), 3.17-2.99 (m, 1H), 2.81-2.65 (m, 2H), 2.54 (s, 3H), 2.43 (s, 3H), 2.24-2.17 (m, 1H), 2.15 (s, 3H).
The title compound was prepared in a similar manner to Example 51, using Intermediate 9b in step a.
m/z ES+ [M+H]+ 674.3; 1H NMR (400 MHz, CD3OD) δ ppm 8.30 (s, 1H), 7.59-7.49 (m, 2H), 7.46-7.36 (m, 1H), 7.22 (s, 1H), 6.42-6.23 (m, 2H), 5.85-5.76 (m, 1H), 4.70 (d, J=9.6 Hz, 1H), 4.67-4.59 (m, 2H), 4.41-4.30 (m, 2H), 4.27-4.17 (m, 1H), 4.15-3.68 (m, 6H), 3.62 (dd, J=14.4, 4.4 Hz, 1H), 3.41 (s, 6H), 3.30-3.22 (m, 1H), 3.17-3.13 (m, 1H), 3.09-2.94 (m, 1H), 2.68 (dd, J=16.8, 8.4 Hz, 1H), 2.54 (s, 3H), 2.38 (dd, J=16.8, 11.6 Hz, 1H).
The title compounds were prepared in a similar manner to Example 51, using tert-butyl octahydro-2H-pyrazino[1,2-a]pyrazine-2-carboxylate (CAS: 1159825-34-9) in step a, and an additional chiral SFC step was conducted after step a.
Example 76a:
m/z ES+ [M+H]+ 658.2; 1H NMR (400 MHz, CDCl3) δ ppm 8.33 (s, 1H), 7.43-7.32 (m, 1H), 7.23 (d, J=8.0 Hz, 1H), 7.16 (t, J=9.2 Hz, 1H), 7.09-7.02 (m, 1H), 6.62-6.44 (m, 1H), 6.30 (d, J=16.4 Hz, 1H), 5.74 (d, J=10.4 Hz, 1H), 4.76-4.35 (m, 3H), 4.03-3.52 (m, 5H), 3.47-3.28 (m, 4H), 3.16-2.64 (m, 7H), 2.57-2.37 (m, 4H), 2.34-1.86 (m, 4H).
Example 76b:
m/z ES+ [M+H]+ 658.3; 1H NMR (400 MHz, CDCl3) δ ppm 8.33 (s, 1H), 7.44-7.34 (m, 1H), 7.27-7.13 (m, 2H), 7.06 (s, 1H), 6.65-6.45 (m, 1H), 6.40-6.23 (m, 1H), 5.87-5.62 (m, 1H), 4.79-4.43 (m, 3H), 4.20-3.57 (m, 6H), 3.40 (m, 4H), 3.10-2.68 (m, 7H), 2.49 (s, 4H), 2.37-2.12 (m, 3H).
Step a. This step was conducted in a similar manner to Example 39, step a, using Intermediate 5b and Intermediate B4.
Step b. A mixture of tert-butyl 3-(3-((3aR,11aS)-6-chloro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)propoxy)azetidine-1-carboxylate (300 mg, 0.46 mmol), methylboronic acid (551 mg, 9.20 mmol), XPhos-Pd-G2 (36.2 mg, 0.046 mmol) and Cs2CO3 (375 mg, 1.15 mmol) in toluene (5 mL) was degassed and purged with N2 3 times. The reaction mixture was stirred at 110° C. for 12 h under a N2 atmosphere. Upon completion, the mixture was filtered, evaporated and purified by reverse phase flash chromatography (water (0.1% TFA)/MeCN) to give tert-butyl 3-(3-((3aR,11aS)-6,10-dimethyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)propoxy)azetidine-1-carboxylate (310 mg, 85% yield) as a white solid.
m/z ES+ [M+H]+ 632.2.
Steps c-d. These 2 steps were conducted in a similar manner to Example 27, steps b-c, using Intermediate E1 in step d.
m/z ES+ [M+H]+ 643.3; 1H NMR (400 MHz, CDCl3) δ ppm 8.33 (s, 1H), 7.27-7.21 (m, 3H), 7.04 (s, 1H), 5.61 (s, 1H), 5.49 (s, 1H), 4.70 (d, J=9.6 Hz, 1H), 4.37-4.33 (m, 1H), 4.25-4.20 (m, 2H), 4.10-4.01 (m, 1H), 3.95-3.79 (m, 1H), 3.60-3.56 (m, 1H), 3.42-3.27 (m, 6H), 3.13-2.94 (m, 4H), 2.84-2.69 (m, 2H), 2.49 (s, 3H), 2.37 (s, 3H), 2.29 (s, 6H), 2.23-2.16 (m, 1H), 1.69-1.62 (m, 2H).
Steps a-b. These 2 steps were conducted in a similar manner to Example 39, steps a-b, using Intermediate 5b and Intermediate B4 in step a.
Step c. This step was conducted in a similar manner to Example 77, step d, using Intermediate E2.
Step d. Acetyl chloride (9 mg, 0.11 mmol) was added to a solution of (3aR,11aS)-6-chloro-5-(3-((1-(2-(hydroxymethyl)acryloyl)azetidin-3-yl)oxy)propyl)-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (40 mg, 0.063 mmol) and TEA (26 mg, 0.25 mmol) in THF (3 mL) at 0° C. The reaction mixture was stirred at 20° C. for 2 h. Upon completion, the mixture was diluted with water (20 mL), extracted with EtOAc (20 mL) and evaporated. Purification by by Prep-TLC (EtOAc) afforded the title compound (14 mg, 33% yield) as a white solid.
m/z ES+ [M+H]+ 678.2; 1H NMR (400 MHz, CDCl3) δ ppm 8.33 (s, 1H), 7.45 (dd, J=7.2, 2.0 Hz, 1H), 7.35-7.28 (m, 2H), 7.05 (s, 1H), 5.64 (s, 1H), 5.50 (s, 1H), 4.77 (s, 2H), 4.63 (d, J=9.2 Hz, 1H), 4.39-4.17 (m, 3H), 4.10-3.78 (m, 2H), 3.62-3.51 (m, 1H), 3.44 (t, J=5.2 Hz, 2H), 3.38 (s, 3H), 3.20-3.07 (m, 2H), 3.05-2.87 (m, 2H), 2.74 (dd, J=16.8, 8.0 Hz, 1H), 2.48 (s, 3H), 2.23 (dd, J=16.8, 11.2 Hz, 1H), 2.10 (s, 3H), 1.64-1.55 (m, 2H).
Steps a-c. These 3 steps were conducted in a similar manner to Example 78, steps a-c, using Intermediate 5a and Intermediate B4 in step a, and Intermediate E3 in step c.
Step d. To a solution of tert-butyl ((Z)-3-chloro-4-(3-(3-((3aR,11aS)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)propoxy)azetidin-1-yl)-4-oxobut-2-en-1-yl)(methyl)carbamate (30 mg, 0.039 mmol) in DCM (0.5 mL) was added TFA (0.1 mL). The mixture was stirred at 25° C. for 1 h. Upon completion, the reaction mixture was evaporated. The residue was diluted with sat. aq. K2CO3 (5 mL) and extracted with EtOAc (3×3 mL). The combined organic layers were washed with brine, dried over Na2SO4, and evaporated to give (3aR,11aS)-5-(3-((1-((Z)-2-chloro-4-(methylamino)but-2-enoyl)azetidin-3-yl)oxy)propyl)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (30 mg, crude) as light yellow oil.
m/z ES+ [M+H]+ 667.0.
Step e. To a solution of (3aR,11aS)-5-(3-((1-((Z)-2-chloro-4-(methylamino)but-2-enoyl)azetidin-3-yl)oxy)propyl)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (25 mg, 0.038 mmol) in MeOH (0.5 mL) was added acetic acid (2 mg, 0.038 mmol), NaBH3CN (5 mg, 0.075 mmol) and formaldehyde (37 wt. % in H2O; 9 mg, 0.11 mmol). The mixture was stirred at 25° C. for 16 h. Upon completion, the reaction mixture was evaporated and the residue was purified by Prep-TLC (EtOAc/MeOH=9/1) followed by Prep-HPLC to give the title compound (1.4 mg, 5% yield) as a yellow solid.
m/z ES+ [M+H]+ 681.2; 1H NMR (400 MHz, CDCl3) δ ppm 8.32 (s, 1H), 7.37-7.31 (m, 1H), 7.22 (d, J=7.6 Hz, 1H), 7.18-7.11 (m, 1H), 7.06 (s, 1H), 6.71 (t, J=6.4 Hz, 1H), 4.62 (d, J=9.2 Hz, 1H), 4.57-4.44 (m, 1H), 4.31-4.13 (m, 3H), 3.97-3.84 (m, 1H), 3.57-3.48 (m, 1H), 3.40 (t, J=6.4 Hz, 2H), 3.35 (s, 3H), 3.24-3.11 (m, 4H), 2.97-2.81 (m, 2H), 2.75-2.69 (m, 1H), 2.50 (s, 3H), 2.30-2.24 (m, 6H), 2.23-2.13 (m, 1H), 1.39-1.24 (m, 2H).
Steps a-c. These 3 steps were conducted in a similar manner to Example 78, steps a-c, using Intermediate 5a and Intermediate B4 in step a, and Intermediate E4 in step c.
Step d. To a mixture of (3aR,11aS)-6-fluoro-5-(3-((1-(3-hydroxy-2-methylenebutanoyl)azetidin-3-yl)oxy)propyl)-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (150 mg, 0.24 mmol) and TEA (479 mg, 4.73 mmol) in DCM (10 mL) was added methanesulfonic anhydride (206 mg, 1.18 mmol). The mixture was stirred at 25° C. for 1 h under a N2 atmosphere. Upon completion, the reaction mixture was diluted with DCM (50 mL) and washed with water (3×15 mL), dried over Na2SO4 and evaporated to give (E)-2-(3-(3-((3aR,11aS)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)propoxy)azetidine-1-carbonyl)but-2-en-1-yl methanesulfonate (150 mg, crude) as a yellow oil.
m/z ES+ [M+H]+ 712.2.
Step e. To a mixture of (E)-2-(3-(3-((3aR,11aS)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)propoxy)azetidine-1-carbonyl)but-2-en-1-yl methanesulfonate (150 mg, 0.21 mmol) in THF (2 mL) was added dimethylamine (2 M in THF, 0.53 mL). The mixture was stirred at 25° C. for 30 min under a N2 atmosphere. Upon completion, the reaction mixture was evaporated and purified by Prep-HPLC to give the title compound (9 mg, 6% yield) as an off-white solid.
m/z ES+ [M+H]+ 661.3; 1H NMR (400 MHz, CDCl3) δ ppm 8.33 (s, 1H), 7.37-7.33 (m, 1H), 7.23 (d, J=8.0 Hz, 1H), 7.19-7.13 (m, 1H), 7.07 (s, 1H), 6.20-6.11 (m, 1H), 4.63 (d, J=8.8 Hz, 1H), 4.38-4.19 (m, 3H), 4.10-3.99 (m, 1H), 3.93-3.80 (m, 1H), 3.59-3.51 (m, 1H), 3.43-3.33 (m, 6H), 3.23-3.14 (m, 3H), 2.97-2.84 (m, 2H), 2.79-2.69 (m, 1H), 2.51 (s, 3H), 2.31-2.17 (m, 7H), 1.83 (d, J=7.2 Hz, 3H), 1.63-1.55 (m, 2H).
Step a. This step was conducted as described in Example 77, step a.
Step b. To a solution of tert-butyl 3-(3-((3aR,11aS)-6-chloro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)propoxy)azetidine-1-carboxylate (90 mg, 0.14 mmol) in MeOH (2 mL) was added 10% Pd/C (20 mg). The reaction mixture was stirred at 40° C. for 12 hr under a H2 atmosphere (15 psi). Upon completion, the reaction mixture was filtered and evaporated to give tert-butyl 3-(3-((3aR,11aS)-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)propoxy)azetidine-1-carboxylate (40 mg, 46% yield) as a white solid.
m/z ES+ [M+H]+ 618.1.
Steps c-d. These 2 steps were conducted in a similar manner to Example 77, steps c-d, using Intermediate E1 in step d.
m/z ES+ [M+H]+ 629.3; 1H NMR (400 MHz, CDCl3) δ ppm 8.31 (s, 1H), 7.47-7.39 (m, 3H), 7.32-7.38 (m, 1H), 7.05 (s, 1H), 5.62 (s, 1H), 5.50 (s, 1H), 4.61 (d, J=8.4 Hz, 1H), 4.50-4.30 (m, 1H), 4.30-4.15 (m, 2H), 4.14-3.96 (m, 1H), 3.96-3.80 (m, 1H), 3.45-3.57 (m, 1H), 3.25-3.44 (m, 6H), 3.05-3.24 (m, 3H), 2.79-2.89 (m, 2H), 2.75-2.65 (m, 1H), 2.51 (s, 3H), 2.43-2.25 (m, 6H), 2.21-2.12 (m, 1H), 1.63-1.53 (m, 2H).
Steps a-d. These 4 steps were conducted in a similar manner to Example 77, steps a-d, using Intermediate 5b and Intermediate B4 in step a, and 1-(tert-butoxycarbonyl)-1,2,5,6-tetrahydropyridine-3-carboxylic acid (CAS: 86447-11-2) in step d.
Step e. To a solution of tert-butyl 5-(3-(3-((3aR,11aS)-6,10-dimethyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)propoxy)azetidine-1-carbonyl)-3,6-dihydropyridine-1(2H)-carboxylate (50 mg, 0.067 mmol) in DCM (1 mL) was added TFA (0.3 mL). The reaction mixture was stirred at 20° C. for 30 min. Upon completion, the mixture was evaporated and the residue was purified by reverse phase flash chromatography (water (0.1% NH4OH)/MeCN) to give the title compound (40 mg, 89% yield) as a yellow solid.
m/z ES+ [M+H]+ 641.2; 1H NMR (400 MHz, CDCl3) δ ppm 8.33 (s, 1H), 7.26-7.18 (m, 3H), 7.05 (s, 1H), 6.32 (s, 1H), 4.70 (d, J=9.6 Hz, 1H), 4.37-4.20 (m, 2H), 3.87-3.71 (m, 2H), 3.59 (dd, J=14.4, 4.0 Hz, 1H), 3.43-3.30 (m, 6H), 3.23-2.96 (m, 9H), 2.86-2.65 (m, 3H), 2.49 (s, 3H), 2.37 (s, 3H), 2.20 (dd, J=16.8, 11.6 Hz, 1H), 1.75-1.55 (m, 2H).
Steps a-b. These 2 steps were conducted in a similar manner to Example 77. Steps a-b, using Intermediate 5b and Intermediate B36 in step a.
Step c. To a solution of (3aR,11aS)-5-(5-(1,3-dioxoisoindolin-2-yl)pentyl)-6,10-dimethyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (140 mg, 0.22 mmol) in EtOH (2.5 mL) was added hydrazine monohydrate (112 mg, 2.21 mmol). The mixture was stirred at 25° C. for 2 h. Upon completion, the reaction mixture was filtered and evaporated to give (3aR,11aS)-5-(5-aminopentyl)-6,10-dimethyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (110 mg, crude) as a yellow solid.
m/z ES+ [M+H]+ 504.0.
Step d. This step was conducted in a similar manner to Example 27, step c.
m/z ES+ [M+H]+ 615.3; 1H NMR (400 MHz, CDCl3) δ ppm 8.34 (s, 1H), 7.25-7.20 (m, 3H), 7.06 (s, 1H), 6.84-6.76 (m, 1H), 6.10-6.00 (m, 2H), 4.67 (d, J=9.2 Hz, 1H), 3.55 (dd, J=14.0, 4.4 Hz, 1H), 3.45-3.36 (m, 1H), 3.35 (s, 3H), 3.24-3.17 (m, 1H), 3.17-3.07 (m, 2H), 3.06-2.98 (m, 1H), 2.96-2.85 (m, 2H), 2.81 (d, J=14.0 Hz, 1H), 2.71 (dd, J=16.8, 8.4 Hz, 1H), 2.51 (s, 3H), 2.37 (s, 3H), 2.32 (s, 6H), 2.18 (dd, J=16.8, 11.6 Hz, 1H), 1.54-1.28 (m, 6H).
The title compound was prepare in a similar manner to Example 83, using Intermediate E1 in step d.
m/z ES+ [M+H]+ 615.3; 1H NMR (400 MHz, CDCl3) δ ppm 9.09 (br s, 1H), 8.33 (s, 1H), 7.25-7.18 (m, 3H), 7.04 (s, 1H), 6.20 (d, J=1.6 Hz, 1H), 5.37 (s, 1H), 4.70 (d, J=9.6 Hz, 1H), 3.58 (dd, J=14.4, 4.4 Hz, 1H), 3.35 (s, 3H), 3.30-3.21 (m, 2H), 3.09 (s, 2H), 3.00-2.85 (m, 3H), 2.82-2.68 (m, 2H), 2.49 (s, 3H), 2.37 (s, 3H), 2.23-2.15 (m, 7H), 1.53-1.45 (m, 2H), 1.44-1.35 (m, 2H), 1.35-1.29 (m, 2H).
The title compound was prepared in a similar manner to Example 83, using Intermediate B35 in step a, and Intermediate E1 in step d.
m/z ES+ [M+H]+ 617.3; 1H NMR (400 MHz, DMSO-d6) δ ppm 8.83-8.70 (m, 1H), 8.17 (s, 1H), 7.35 (s, 1H), 7.32-7.21 (m, 3H), 5.86 (d, J=2.0 Hz, 1H), 5.39 (s, 1H), 4.62 (d, J=9.2 Hz, 1H), 3.55-3.47 (m, 1H), 3.39-3.36 (m, 2H), 3.32-3.29 (m, 2H), 3.29-3.24 (m, 2H), 3.24 (s, 3H), 3.11-3.05 (m, 1H), 3.04 (s, 2H), 2.93-2.74 (m, 3H), 2.59-2.55 (m, 1H), 2.48 (s, 3H), 2.42-2.36 (m, 1H), 2.33 (s, 3H), 2.10 (s, 6H).
Steps a-c. These 3 steps were conducted in a similar manner to Intermediate 13, steps a-c, using Intermediate 5b in step a.
Step d. To a solution of 3-((3aR,11aS)-6-chloro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)propanal (360 mg, 0.73 mmol), NaH2PO4 (261 mg, 2.18 mmol) and 2-methylbut-2-ene (714 mg, 10.1 mmol) in tert-butanol (4 mL) and H2O (4 mL) was added sodium chlorite (197 mg, 2.18 mmol). The mixture was stirred at 20° C. for 1 h. Upon completion, the reaction mixture was acidified to pH 5 with the slow addition of 0.5 M HCl. The aqueous mixture was extracted with EtOAc (3×10 mL). The combined organic layers were washed with brine (30 mL), dried over Na2SO4 and evaporated. The residue was purified by reverse phase flash chromatography (water (0.1% FA)/MeCN) to give 3-((3aR,11aS)-6-chloro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)propanoic acid (160 mg, 43% yield) as a white solid.
m/z ES+ [M+H]+ 511.0.
Step e. HATU (167 mg, 0.44 mmol) was added to a solution of 3-((3aR,11aS)-6-chloro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)propanoic acid (150 mg, 0.29 mmol), tert-butyl piperazine-1-carboxylate (109 mg, 0.59 mmol) and DIPEA (76 mg, 0.59 mmol) in DCM (3 mL) at 20° C. The reaction mixture was stirred for 1 h. Upon completion, the mixture was evaporated and the residue was purified by reverse phase flash chromatography (water (0.1% NH4OH)/MeCN) to give tert-butyl 4-(3-((3aR,11aS)-6-chloro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)propanoyl)piperazine-1-carboxylate (90 mg, 45% yield) as a yellow oil.
m/z ES+ [M+H]+ 679.3.
Steps f-g. These 2 steps were conducted in a similar manner to Example 77, steps c-d.
m/z ES+ [M+H]+ 690.2; 1H NMR (400 MHz, CDCl3) δ ppm 8.32 (s, 1H), 7.47-7.43 (m, 1H), 7.36-7.28 (m, 2H), 7.05 (s, 1H), 6.19 (s, 1H), 5.73 (s, 1H), 4.58 (d, J=9.2 Hz, 1H), 3.88 (s, 2H), 3.82-3.61 (m, 7H), 3.55 (s, 2H), 3.50-3.40 (m, 2H), 3.38 (s, 3H), 3.03-2.92 (m, 1H), 2.84 (s, 6H), 2.75 (dd, J=16.8, 8.4 Hz, 1H), 2.48 (s, 3H), 2.47-2.28 (m, 2H), 2.25-2.01 (m, 2H).
Step a. This step was conducted in a similar manner to Example 39, step a, using Intermediate 5b and 4-nitrobenzaldehyde (CAS: 555-16-8).
Step b. This step was conducted in a similar manner to Example 77, step b.
Step c. To a solution of (3aR,11aS)-6,10-dimethyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-5-(4-nitrobenzyl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (100 mg, 0.18 mmol) in EtOAc (3 mL) was added 3% Pt—V/C (50 mg, 0.006 mmol). The reaction mixture was stirred at 40° C. for 12 hr under a H2 atmosphere (15 psi). Upon completion, the reaction mixture was filtered and evaporated to give (3aR,11aS)-5-(4-aminobenzyl)-6,10-dimethyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (80 mg, 85% yield) as a yellow gum.
m/z ES+ [M+H]+ 524.2.
Step d. This step was conducted in a similar manner to Example 27, step c, using acrylic acid.
m/z ES+ [M+H]+ 578.2; 1H NMR (400 MHz, CDCl3) δ ppm 8.34 (s, 1H), 7.71 (s, 1H), 7.60 (d, J=7.6 Hz, 2H), 7.34 (d, J=8.4 Hz, 2H), 7.31-7.27 (m, 2H), 7.27-7.24 (m, 1H), 7.08 (s, 1H), 6.51-6.40 (m, 1H), 6.31-6.19 (m, 1H), 5.76 (d, J=10.4 Hz, 1H), 4.78 (d, J=9.6 Hz, 1H), 4.15 (d, J=13.6 Hz, 1H), 3.99 (d, J=13.6 Hz, 1H), 3.49-3.39 (m, 4H), 3.07-2.91 (m, 1H), 2.79-2.68 (m, 1H), 2.62 (dd, J=16.8, 8.8 Hz, 1H), 2.53 (s, 3H), 2.49 (s, 3H), 2.20-2.10 (m, 1H).
Steps a-d. These 4 steps were conducted in a similar manner to Example 87, steps a-d, using Intermediate 5b and 4-nitrobenzaldehyde (CAS: 555-16-8) in step a, 1-(tert-butoxycarbonyl)-1,2,5,6-tetrahydropyridine-3-carboxylic acid (CAS: 86447-11-2) in step d.
Step e. This step was conducted in a similar manner to Example 82, step e.
m/z ES+ [M+H]+ 633.2; 1H NMR (400 MHz, CDCl3) δ ppm 8.31 (s, 1H), 7.87 (br s, 1H), 7.58-7.48 (m, 2H), 7.36-7.28 (m, 2H), 7.27-7.19 (m, 3H), 7.04 (s, 1H), 6.72 (s, 1H), 4.78-4.69 (m, 1H), 4.18-4.06 (m, 1H), 3.98-3.95 (m, 1H), 3.78-3.71 (m, 1H), 3.47-3.31 (m, 4H), 3.08 2.91 (m, 2H), 2.75-2.57 (m, 2H), 2.50 (s, 3H), 2.46 (s, 3H), 2.39-2.31 (m, 2H), 2.20-2.08 (m, 4H).
Steps a-c. These 3 steps were conducted as described in Example 87, steps a-c.
Step d. A mixture of (3aR,11aS)-5-(4-aminobenzyl)-6,10-dimethyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (100 mg, 0.19 mmol), Intermediate E6 (113 mg, 0.57 mmol) and TEA (193 mg, 1.91 mmol) in DCM (1 mL) was degassed and purged with N2 3 times. The reaction mixture was stirred at 25° C. for 1 h under a N2 atmosphere. Upon completion, the reaction mixture was filtered and evaporated. The residue was purified by Prep-HPLC to give the title compound (19 mg, 15% yield) as a white solid.
m/z ES+ [M+H]+ 649.3; 1H NMR (400 MHz, CDCl3) δ ppm 11.55 (s, 1H), 8.34 (s, 1H), 7.57 (d, J=8.4 Hz, 2H), 7.31 (d, J=8.4 Hz, 2H), 7.28-7.23 (m, 3H), 7.12-7.04 (m, 2H), 4.77 (d, J=9.6 Hz, 1H), 4.17-4.09 (m, 1H), 4.00-3.94 (m, 1H), 3.48-3.41 (m, 4H), 3.25 (s, 2H), 3.09-2.94 (m, 1H), 2.75-2.61 (m, 2H), 2.52 (s, 3H), 2.48 (s, 3H), 2.37 (s, 6H), 2.14 (dd, J=16.8, 11.6 Hz, 1H), 1.89 (d, J=7.2 Hz, 3H).
The title compound was prepared in a similar manner to Example 89, using Intermediate B37 in step a, and acryloyl chloride in step d.
m/z ES+ [M+H]+ 608.1; 1H NMR (400 MHz, CDCl3) δ ppm 8.41 (d, J=8.0 Hz, 1H), 8.33 (s, 1H), 7.87 (s, 1H), 7.27-7.24 (s, 3H), 7.05 (s, 1H), 6.97-6.90 (m, 2H), 6.47-6.39 (m, 1H), 6.34-6.25 (m, 1H), 5.77 (dd, J=10.0, 1.2 Hz, 1H), 4.74 (d, J=9.6 Hz, 1H), 4.15-4.02 (m, 2H), 3.93 (s, 3H), 3.46 (dd, J=14.0, 4.8 Hz, 1H), 3.40 (s, 3H), 3.00-2.87 (m, 1H), 2.75 (dd, J=14.0, 11.6 Hz, 1H), 2.64 (dd, J=16.8, 8.4 Hz, 1H), 2.50 (s, 6H), 2.13 (dd, J=16.8, 11.6 Hz, 1H).
Step a. This step was conducted in a similar manner to Example 39, step a, using Intermediate 5b and 3-fluoro-4-nitrobenzaldehyde (CAS: 160538-51-2).
Step b. To a solution of (3aR,11aS)-6-chloro-5-(3-fluoro-4-nitrobenzyl)-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (180 mg, 0.30 mmol) in MeCN (10 mL) was added K2CO3 (126 mg, 0.91 mmol) and N1,N1,N2-trimethylethane-1,2-diamine (CAS: 142-25-6; 37 mg, 0.37 mmol). The mixture was stirred at 40° C. for 3 h. Upon completion, the reaction mixture was diluted with H2O (10 mL) and extracted with EtOAc (3×5 mL). The combined organic layers were dried over Na2SO4, filtered and evaporated. The residue was purified by reverse phase flash chromatography (water (0.1% FA)/MeCN) to give (3aR,11aS)-6-chloro-5-(3-((2-(dimethylamino)ethyl)(methyl)amino)-4-nitrobenzyl)-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (200 mg, 96% yield) as red oil.
m/z ES+ [M+H]+ 674.1.
Steps c-e. These 3 steps were conducted in a similar manner to Example 89, steps b-d, using acryloyl chloride in step d.
m/z ES+ [M+H]+ 678.4; 1H NMR (400 MHz, CDCl3) δ ppm 10.11 (br s, 1H), 8.44 (s, 1H), 8.33 (s, 1H), 7.27-7.22 (m, 4H), 7.16 (s, 1H), 7.11 (d, J=8.4 Hz, 1H), 7.05 (s, 1H), 6.45 (d, J=16.0 Hz, 1H), 5.73 (d, J=11.6 Hz, 1H), 4.73 (d, J=9.2 Hz, 1H), 4.12-4.01 (m, 2H), 3.48-3.42 (m, 1H), 3.38 (s, 3H), 2.97-2.84 (m, 2H), 2.76 (d, J=13.6 Hz, 1H), 2.72 (s, 4H), 2.67-2.58 (m, 2H), 2.51 (s, 4H), 2.48 (s, 4H), 2.43-2.25 (m, 4H), 2.20-2.10 (m, 2H).
Steps a-b. These 2 steps were conducted in a similar manner to Example 39, steps a-b, using Intermediate 5b and tert-butyl 4-formylpiperidine-1-carboxylate (CAS: 137076-22-3) in step a.
Step c. To a solution of (3aR,11aS)-6-chloro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-5-(piperidin-4-ylmethyl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (300 mg, 0.56 mmol) and tert-butyl 3-oxoazetidine-1-carboxylate (115 mg, 0.67 mmol) in MeOH (5 mL) was added 4 Å molecular sieves (100 mg) and acetic acid (3 mg, 0.056 mmol). The mixture was stirred at 20° C. for 30 min and then NaBH3CN (70 mg, 1.12 mmol) was added. The reaction mixture was stirred at 50° C. for 15 h 30 min. Upon completion, the reaction mixture was filtered and evaporated. Sat. aq. NaHCO3 (10 mL) was added to the residue and the aqueous mixture was further diluted with water (10 mL) and extracted with EtOAc (3×20 mL). The combined organic layers were washed with brine (3×20 mL), dried over Na2SO4 and evaporated. The crude product was purified by column chromatography (EtOAc) to give tert-butyl 3-(4-(((3aR,11aS)-6-chloro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)methyl)piperidin-1-yl)azetidine-1-carboxylate (240 mg, 62% yield) as a white solid.
m/z ES+ [M+H]+ 691.4; 1H NMR (400 MHz, CDCl3) δ ppm 8.32 (s, 1H), 7.44 (dd, J=7.2, 2.4 Hz, 1H), 7.33-7.27 (m, 2H), 7.05 (s, 1H), 4.62 (d, J=9.2 Hz, 1H), 3.90 (m, 2H), 3.80 (m, 2H), 3.51 (dd, J=14.0, 4.4 Hz, 1H), 3.36 (s, 3H), 3.07-2.67 (m, 8H), 2.48 (s, 3H), 2.22 (dd, J=16.8, 11.2 Hz, 1H), 1.93-1.65 (m, 4H), 1.42 (s, 9H), 1.35-1.29 (m, 1H), 1.23-0.99 (m, 2H).
Steps d-e. These 2 steps were conducted in a similar manner to Example 38, steps b-c.
m/z ES+ [M+H]+ 645.1; 1H NMR (400 MHz, CDCl3) δ ppm 8.32 (s, 1H), 7.45 (dd, J=7.2, 2.4 Hz, 1H), 7.33-7.28 (m, 2H), 7.05 (s, 1H), 6.37-6.27 (m, 1H), 6.23-6.11 (m, 1H), 5.66 (dd, J=10.4, 1.6 Hz, 1H), 4.62 (d, J=9.2 Hz, 1H), 4.26-4.17 (m, 1H), 4.15-3.68 (m, 3H), 3.51 (dd, J=14.0, 4.0 Hz, 1H), 3.41-3.30 (m, 3H), 3.25-3.12 (m, 1H), 3.07-2.79 (m, 6H), 2.73 (dd, J=16.8, 8.4 Hz, 1H), 2.48 (s, 3H), 2.22 (dd, J=16.8, 11.2 Hz, 1H), 1.90-1.81 (m, 3H), 1.39-1.13 (m, 4H).
Steps a-c. These 3 steps were conducted as described in Example 92, steps a-c.
Step d. This step was conducted in a similar manner to Example 77, step b.
Steps e-f. These 2 steps were conducted in a similar manner to Example 92, steps d-e.
m/z ES+ [M+H]+ 625.4; 1H NMR (400 MHz, CDCl3) δ ppm 8.31 (s, 1H), 7.27-7.19 (m, 3H), 7.05 (s, 1H), 6.33 (dd, J=16.8, 1.6 Hz, 1H), 6.20-6.09 (m, 1H), 5.72 (dd, J=10.4, 1.2 Hz, 1H), 4.97-4.71 (m, 1H), 4.68 (d, J=9.2 Hz, 1H), 4.58-4.18 (m, 3H), 3.84-3.65 (m, 1H), 3.63-3.27 (m, 5H), 3.04-2.79 (m, 4H), 2.72 (dd, J=16.8, 8.4 Hz, 1H), 2.60-2.48 (m, 4H), 2.37 (s, 3H), 2.20 (dd, J=16.8, 11.2 Hz, 1H), 2.09-1.55 (m, 7H).
Step a. This step was conducted in a similar manner to Example 92, step a, using Intermediate 5b and Intermediate B38.
Step b. This step was conducted in a similar manner to Example 77, step b.
Steps c-f. These 4 steps were conducted in a similar manner to Example 92, steps b-e.
m/z ES+ [M+H]+ 643.1; 1H NMR (400 MHz, CDCl3) δ ppm 8.31 (s, 1H), 7.27-7.22 (s, 3H), 7.04 (s, 1H), 6.38-6.25 (m, 1H), 6.23-6.11 (m, 1H), 5.66 (d, J=10.4 Hz, 1H), 4.66 (d, J=9.6 Hz, 1H), 4.25-4.17 (m, 1H), 4.15-3.98 (m, 2H), 3.94-3.84 (m, 1H), 3.73-3.61 (m, 1H), 3.35 (s, 3H), 3.23-3.08 (m, 2H), 3.07-2.91 (m, 2H), 2.86-2.75 (m, 1H), 2.72-2.65 (m, 1H), 2.63-2.55 (m, 2H), 2.49 (s, 3H), 2.43 (s, 3H), 2.22-2.12 (m, 3H), 2.01-1.92 (m, 2H), 1.66-1.40 (m, 2H).
The title compounds were prepared in a similar manner to Example 94, using Intermediate 5b and tert-butyl 4-formylpiperidine-1-carboxylate (CAS: 137076-22-3) in step a, and tert-butyl 3-oxopyrrolidine-1-carboxylate (CAS: 101385-93-7) in step d. An additional chiral SFC step was conducted after step d.
Example 95a:
m/z ES+ [M+H]+ 639.2; 1H NMR (400 MHz, CDCl3) δ ppm 8.32 (s, 1H), 7.26-7.19 (m, 3H), 7.05 (s, 1H), 6.47-6.33 (m, 2H), 5.75-5.65 (m, 1H), 4.69 (d, J=9.2 Hz, 1H), 4.05-3.67 (m, 2H), 3.61-3.41 (m, 2H), 3.35 (s, 3H), 3.06-2.67 (m, 7H), 2.49 (s, 3H), 2.39 (s, 3H), 2.24-2.16 (m, 2H), 2.12-1.89 (m, 2H), 1.75-1.55 (m, 5H), 1.48-1.06 (m, 3H).
Example 95b:
m/z ES+ [M+H]+ 639.3; 1H NMR (400 MHz, CDCl3) δ ppm 8.32 (s, 1H), 7.25-7.19 (m, 3H), 7.04 (s, 1H), 6.45-6.32 (m, 2H), 5.72-5.65 (m, 1H), 4.71-4.66 (m, 1H), 3.98-3.71 (m, 2H), 3.59-3.45 (m, 2H), 3.45-3.31 (m, 4H), 3.11-2.66 (m, 8H), 2.49 (s, 3H), 2.39 (s, 3H), 2.29-1.88 (m, 5H), 1.85-1.56 (m, 4H), 1.30-1.19 (m, 1H).
Steps a-e. These 5 steps were conducted in a similar manner to Example 92, steps a-e, using Intermediate 5b and Intermediate B39 in step a.
Step f. A mixture of (3aR,11aS)-5-((1-(1-acryloylazetidin-3-yl)-3-((tert-butyldiphenylsilyl)oxy)piperidin-4-yl)methyl)-6-chloro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (20 mg, 0.022 mmol) and TBAF (1 M in THF, 0.13 mL) in THF (1 mL) was stirred at 15° C. for 12 h. Upon completion, the mixture was quenched with water (1 mL), further diluted with water (30 mL) and extracted with EtOAc (2×30 mL). The combined organic layers were evaporated and the residue was purified by reverse phase flash chromatography (water (0.1% FA)/MeCN) followed by Prep-HPLC to give the title compound (0.7 mg, 5% yield) as a white solid.
m/z ES+ [M+H]+ 661.1; 1H NMR (400 MHz, CDCl3) δ ppm 8.34 (s, 1H), 7.48 (dd, J=6.8, 2.8 Hz, 1H), 7.35-7.29 (m, 2H), 7.05 (s, 1H), 6.41-6.28 (m, 1H), 6.25-6.13 (m, 1H), 5.67 (d, J=11.6 Hz, 1H), 4.62 (d, J=8.8 Hz, 1H), 4.29-3.86 (m, 4H), 3.66-3.50 (m, 2H), 3.40 (s, 3H), 3.34-3.17 (m, 2H), 3.12-2.70 (m, 7H), 2.47 (s, 3H), 2.31-2.19 (m, 1H), 1.89-1.82 (m, 3H), 1.38-1.28 (m, 2H).
Step a. This step was conducted in a similar manner to Example 92, step a, using Intermediate 5a and Intermediate B46 in step a.
Step b. To a solution of benzyl 4-(1-((3aR,11aS)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)propan-2-yl)piperazine-1-carboxylate (50 mg, 0.073 mmol) in MeOH (2 mL) was added 10% Pd/C (50 mg). The mixture was stirred at 30° C. for 16 h under a H2 atmosphere (15 psi). Upon completion, the reaction mixture was filtered and evaporated to give (3aR,11aS)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-5-(2-(piperazin-1-yl)propyl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (34 mg, crude) as white solid.
m/z ES+ [M+H]+ 549.3.
Steps c-e. These 3 steps were conducted in a similar manner Example 92, steps c-e.
m/z ES+ [M+H]+ 658.3; 1H NMR (400 MHz, CDCl3) δ ppm 8.32 (s, 1H), 7.41-7.29 (m, 1H), 7.26-7.22 (m, 1H), 7.19-7.13 (m, 1H), 7.06 (s, 1H), 6.37-6.27 (m, 1H), 6.22-6.12 (m, 1H), 5.69-5.65 (m, 1H), 4.61-4.56 (m, 1H), 4.26-4.17 (m, 1H), 4.15-3.99 (m, 2H), 3.94-3.83 (m, 1H), 3.62-3.55 (m, 1H), 3.37 (s, 3H), 3.30-3.19 (m, 1H), 3.09-2.80 (m, 5H), 2.78-2.51 (m, 5H), 2.50 (s, 3H), 2.25-2.15 (m, 1H), 1.80-1.41 (m, 4H), 1.37-0.88 (m, 3H).
Steps a-b. These 2 steps were conducted as described in Intermediate 11a and Intermediate 11b, steps a-b.
Step c. This step was conducted in a similar manner to Example 38, step b.
Step d. To a mixture of (3aR,11aS)-5-((5-(azetidin-3-yl)-1H-1,2,4-triazol-3-yl)methyl)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (30 mg, 0.045 mmol, TFA salt), acrylic acid (3 mg, 0.045 mmol) and DIPEA (17 mg, 0.13 mmol) in DCM (2 mL) was added HATU (20 mg, 0.054 mmol). The reaction mixture was stirred at 20° C. for 30 min. Upon completion, the reaction mixture was evaporated and the residue was purified by Prep-TLC (DCM/MeOH=20/1) followed by Prep-HPLC to give the title compound (12 mg, 43% yield) as a white solid.
m/z ES+ [M+H]+ 613.1; 1H NMR (400 MHz, CDCl3) δ ppm 8.32 (s, 1H), 7.45-7.36 (m, 1H), 7.26-7.17 (m, 2H), 7.08 (s, 1H), 6.38-6.30 (m, 1H), 6.26-6.15 (m, 1H), 5.74-5.65 (m, 1H), 4.66 (d, J=9.6 Hz, 1H), 4.59-4.37 (m, 5H), 4.32-4.20 (m, 1H), 4.04-3.93 (m, 1H), 3.71-3.60 (m, 1H), 3.33 (s, 3H), 3.12-3.01 (m, 1H), 2.99-2.87 (m, 1H), 2.78-2.68 (m, 1H), 2.51 (s, 3H), 2.27-2.17 (m, 1H).
Steps a-b. These 2 steps were conducted in a similar manner to Example 38, steps b-c, using Intermediate 11a in step a.
m/z ES+ [M+H]+ 627.3; 1H NMR (400 MHz, CDCl3) δ ppm 8.30 (s, 1H), 7.42-7.34 (m, 1H), 7.23-7.13 (m, 2H), 7.06 (s, 1H), 6.40-6.30 (m, 1H), 6.25-6.16 (m, 1H), 5.68 (d, J=10.4 Hz, 1H), 4.61 (dd, J=9.6, 1.6 Hz, 1H), 4.53-4.47 (m, 1H), 4.47-4.41 (m, 2H), 4.40-4.32 (m, 2H), 4.13-4.06 (m, 1H), 3.90-3.85 (m, 1H), 3.84 (d, J=4.0 Hz, 3H), 3.66-3.56 (m, 1H), 3.26 (d, J=13.6 Hz, 3H), 3.06-2.93 (m, 1H), 2.79 (dd, J=9.6, 4.8 Hz, 1H), 2.74-2.66 (m, 1H), 2.48 (s, 3H), 2.20 (dd, J=16.8, 11.6 Hz, 1H).
The title compound was prepared in a similar manner to Example 99, using Intermediate 11b in step a.
m/z ES+ [M+H]+ 627.3; 1H NMR (400 MHz, CDCl3) δ ppm 8.30 (s, 1H), 7.36-7.28 (m, 1H), 7.20 (d, J=8.4 Hz, 1H), 7.17-7.10 (m, 1H), 7.04 (s, 1H), 6.41-6.30 (m, 1H), 6.26-6.16 (m, 1H), 5.70 (t, J=9.6 Hz, 1H), 4.73-4.53 (m, 3H), 4.50-4.41 (m, 1H), 4.39-4.31 (m, 1H), 4.26-4.08 (m, 2H), 3.94-3.74 (m, 2H), 3.72 (d, J=4.4 Hz, 3H), 3.35-3.21 (m, 3H), 3.05-2.75 (m, 2H), 2.72-2.63 (m, 1H), 2.48 (s, 3H), 2.26-2.11 (m, 1H).
Step a. This step was conducted as described in Intermediate 11a and Intermediate 11b, step a.
Step b. A mixture of 2-((3aR,11aS)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)acetonitrile (300 mg, 0.65 mmol), hydroxylamine hydrochloride (90 mg, 1.30 mmol) and K2CO3 (269 mg, 1.95 mmol) in EtOH (3 mL) was stirred at 100° C. for 16 hr. Upon completion, the reaction mixture was diluted with water (10 ml) and extracted with EtOAc (2×10 ml). The combined organic layers were evaporated and the residue was purified by Prep-TLC (EtOAc) to give 2-((3aR,11aS)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)-N′-hydroxyacetimidamide (150 mg, 46% yield) as a white solid.
1H NMR (400 MHz, CDCl3) δ ppm 8.34 (s, 1H), 7.46-7.36 (m, 1H), 7.26 (d, J=8.4 Hz, 1H), 7.23-7.16 (m, 1H), 7.08 (s, 1H), 5.57 (br s, 2H), 4.73-4.60 (m, 1H), 4.03-3.69 (m, 3H), 3.63-3.54 (m, 1H), 3.44-3.38 (m, 3H), 3.07-2.70 (m, 3H), 2.51 (s, 3H), 2.28-2.13 (m, 1H).
Step c. To a solution of 1-(tert-butoxycarbonyl)azetidine-3-carboxylic acid (63 mg, 0.32 mmol) in DMF (1 mL) was added CDI (79 mg, 0.49 mmol). The reaction mixture was stirred at 25° C. for 6 h, after which, 2-((3aR,11aS)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)-N′-hydroxyacetimidamide (120 mg, 0.24 mmol) was added and the reaction mixture was stirred at 25° C. for a further 10 h. Upon completion, the reaction mixture was concentrated to give tert-butyl 3-((2-((3aR,11aS)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)-1-(hydroxyimino)ethyl)carbamoyl)azetidine-1-carboxylate (140 mg, crude) as a yellow oil.
m/z ES+ [M+H]+ 678.0.
Step d. To a solution of tert-butyl 3-((2-((3aR,11aS)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)-1-(hydroxyimino)ethyl)carbamoyl)azetidine-1-carboxylate (140 mg, 0.21 mmol) in 1,4-dioxane (1 mL) was added DBU (63 mg, 0.41 mmol) and the mixture was stirred at 25° C. for 3 h. Upon completion, the reaction mixture was evaporated and the residue was purified by column chromatography (EtOAc) to give tert-butyl 3-(3-(((3aR,11aS)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)methyl)-1,2,4-oxadiazol-5-yl)azetidine-1-carboxylate (120 mg, 79% yield) as a white solid.
m/z ES+ [M+H]+ 660.1.
Steps e-f. These 2 steps were conducted in a similar manner to Example 98, steps c-d.
m/z ES+ [M+H]+ 614.1; 1H NMR (400 MHz, CDCl3) δ ppm 8.33 (s, 1H), 7.43-7.36 (m, 1H), 7.27-7.16 (m, 2H), 7.07 (s, 1H), 6.45-6.35 (m, 1H), 6.28-6.17 (m, 1H), 5.79-5.70 (m, 1H), 4.68-4.62 (m, 2H), 4.61-4.42 (m, 3H), 4.37-4.21 (m, 2H), 4.12-4.07 (m, 1H), 3.83-3.72 (m, 1H), 3.32 (s, 3H), 3.12-3.02 (m, 1H), 3.00-2.79 (m, 1H), 2.78-2.69 (m, 1H), 2.50 (s, 3H), 2.27-2.14 (m, 1H).
Step a. To a solution of Intermediate 9a (170 mg, 0.35 mmol) in DMF (2 mL) was added CDI (207 mg, 1.27 mmol) and the mixture was stirred at 25° C. for 6 h, after which, tert-butyl 3-(N′-hydroxycarbamimidoyl)azetidine-1-carboxylate (CAS: 1309207-05-3; 91 mg, 0.43 mmol) was added. The reaction mixture was stirred at 25° C. for a further 10 h. Upon completion, the mixture was evaporated to give tert-butyl 3-(N-(2-((3aR,11aS)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)acetyl)-N′-hydroxycarbamimidoyl)azetidine-1-carboxylate (240 mg, crude) as a white solid.
m/z ES+ [M+H]+ 678.2.
Step b. This step was conducted in a similar manner to Example 101, step d.
Steps c-d. These 2 steps were conducted in a similar manner to Example 38, steps b-c.
m/z ES+ [M+H]+ 614.2; 1H NMR (400 MHz, CDCl3) δ ppm 8.32 (s, 1H), 7.43-7.36 (m, 1H), 7.27-7.23 (m, 1H), 7.22-7.15 (m, 1H), 7.06 (s, 1H), 6.41-6.32 (m, 1H), 6.26-6.15 (m, 1H), 5.75-5.67 (m, 1H), 4.63 (d, J=9.6 Hz, 1H), 4.60-4.54 (m, 1H), 4.52 (s, 2H), 4.49-4.41 (m, 2H), 4.26-4.14 (m, 1H), 4.02-3.98 (m, 1H), 3.80-3.69 (m, 1H), 3.32 (d, J=17.2 Hz, 3H), 3.16-3.06 (m, 1H), 3.03-2.82 (m, 1H), 2.75 (dd, J=16.8, 8.4 Hz, 1H), 2.49 (s, 3H), 2.22 (dd, J=16.8, 11.6 Hz, 1H).
Step a. To a solution of Intermediate 9a (200 mg, 0.42 mmol) and tert-butyl 3-(hydrazinecarbonyl)azetidine-1-carboxylate (CAS: 1001907-44-3; 107 mg, 0.5 mmol) in DCM (4 mL) was added EDCI (96 mg, 0.5 mmol) and HOBt (73 mg, 0.54 mmol). The mixture was stirred at 25° C. for 5 h. Upon completion, the reaction mixture was diluted in water (10 mL) and extracted with EtOAc (5 mL). The aqueous layer was further extracted with EtOAc (2×10 mL). The combined organic layers were dried over Na2SO4 and evaporated. The residue was purified by column chromatography (EtOAc/MeOH=20/1) to give tert-butyl 3-(2-(2-((3aR,11aS)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)acetyl)hydrazine-1-carbonyl)azetidine-1-carboxylate (135 mg, 46% yield) as a white solid.
m/z ES+ [M+H]+ 678.1; 1H NMR (400 MHz, CDCl3) δ ppm 9.12 (d, J=3.2 Hz, 1H), 8.33 (s, 1H), 7.95 (d, J=4.8 Hz, 1H), 7.43-7.40 (m, 1H), 7.27-7.19 (m, 2H), 7.08 (s, 1H), 4.73-4.68 (m, 1H), 4.16-4.06 (m, 4H), 3.83 (s, 2H), 3.67-3.64 (m, 1H), 3.43 (s, 3H), 3.30-3.22 (m, 1H), 3.11-3.01 (m, 2H), 2.85-2.74 (m, 1H), 2.50 (s, 3H), 2.28-2.23 (m, 1H), 1.44 (s, 9H).
Step b. To a solution of tert-butyl 3-(2-(2-((3aR,11aS)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)acetyl)hydrazine-1-carbonyl)azetidine-1-carboxylate (100 mg, 0.15 mmol) in MeCN (1 mL) was added POCl3 (68 mg, 0.44 mmol). The mixture was stirred at 80° C. for 4 h. Upon completion, the reaction mixture was quenched with sat. aq. NaHCO3 (20 mL). The aqueous mixture was extracted with EtOAc (3×25 mL). The combined organic layers were washed with brine (50 mL), dried over Na2SO4 and evaporated to give (3aR,11aS)-5-((5-(azetidin-3-yl)-1,3,4-oxadiazol-2-yl)methyl)-6-fluoro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (100 mg, crude) was obtained as a yellow solid.
m/z ES+ [M+H]+ 560.2.
Step c. This step was conducted in a similar manner to Example 38, step c.
m/z ES+ [M+H]+ 614.2; 1H NMR (400 MHz, CDCl3) δ ppm 8.32 (s, 1H), 7.45-7.30 (m, 2H), 7.24-7.17 (m, 1H), 7.07 (s, 1H), 6.44-6.32 (m, 1H), 6.30-6.18 (m, 1H), 5.75-5.70 (m, 1H), 4.71-4.57 (m, 4H), 4.53-4.46 (m, 1H), 4.40-4.22 (m, 2H), 4.19-4.05 (m, 1H), 3.82-3.69 (m, 1H), 3.27-3.21 (m, 3H), 3.13-3.00 (m, 1H), 2.79-2.58 (m, 2H), 2.49 (s, 3H), 2.27-2.12 (m, 1H).
The Examples in Table 9 were prepared using methods similar to those described in the synthesis of Example 1, using the listed Intermediates in step a.
1H NMR (400 MHz) δ ppm
The Examples in Table 10 were prepared using methods similar to those described in the synthesis of Example 1, using the listed Intermediates in step a, and an additional chiral SFC step was conducted.
1H NMR (400 MHz) δ ppm
The Examples in Table 11 were prepared using methods similar to those described in the synthesis of Example 38 or Example 39, using the listed Intermediates in step a, and the appropriate warhead in step c.
1H NMR (400 MHz) δ ppm
The Examples in Table 12 were prepared using methods similar to those described in the synthesis of Example 38 or Example 39, using the listed Intermediates in step a, and an additional chiral SFC step was conducted.
1H NMR (400 MHz) δ ppm
The Examples in Table 13 were prepared using methods similar to those described in the synthesis of Example 39 or Example 43, using the listed Intermediates in step a, and the appropriate warhead in step c.
1H NMR (400 MHz) δ ppm
The Examples in Table 14 were prepared using methods similar to those described in the synthesis of Example 39 or Example 43, using the listed Intermediates in step a, and the appropriate warhead in step c. An additional chiral SFC step was conducted after step a.
1H NMR (400 MHz)
The Examples in Table 15 were prepared using methods similar to those described in the synthesis of Example 77, using the listed Intermediates in step a, and the appropriate warhead in step d. For Example 199, an additional chiral SFC step was conducted after step b.
1H NMR (400 MHz)
The Examples in Table 16 were prepared using methods similar to those described in the synthesis of Example 77, using the listed Intermediates in step a. The final step was conducted in a similar manner to the amide coupling with acryloyl chloride described for Example 1. For Example 206a and Example 206b an additional chiral SFC step was conducted after step b.
1H NMR (400 MHz) δ ppm
The Examples in Table 17 were prepared using methods similar to those described in the synthesis of Example 87, using the listed Intermediates in step a, and the appropriate warhead in step d.
1H NMR (400 MHz) δ ppm
The Examples in Table 18 were prepared using methods similar to those described in the synthesis of Example 92, using the listed Intermediates in step a, and the appropriate ketone in step c.
1H NMR (400 MHz) δ ppm
The Examples in Table 19 were prepared using methods similar to those described in the synthesis of Example 92, using the listed Intermediates in step a, and the appropriate ketone in step c. An additional chiral SFC step was conducted after step c.
1H NMR (400 MHz) δ ppm
Step a. To a solution of Intermediate 5b (7.4 g, 16.7 mmol) and Intermediate B49 (6.21 g, 21.9 mmol) in DMF (100 mL) was added TMSCI (4.58 g, 42.2 mmol). The mixture was stirred at 0° C. for 30 min. A solution of borane tetrahydrofuran complex (1 M in THF, 18.5 mL) in THF (186 mL) was added and the reaction mixture was stirred at 20° C. for 11 h 30 min. Upon completion, the reaction mixture was quenched with sat. aq. NaHCO3 (50 mL) and H2O (300 mL). The aqueous mixture was extracted with EtOAc (3×300 mL). The combined organic layers were washed with brine (500 mL), dried over Na2SO4 and evaporated. The residue was purified by prep-TLC (PE/EtOAc=1/3) followed by reverse phase flash chromatography (water (0.1% NH4OH)/MeCN) to give benzyl 2-(((3aR,11aS)-6-chloro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)methyl)-5,7-dihydro-6H-pyrrolo[3,4-b]pyrazine-6-carboxylate (3.7 g, 31% yield) as a yellow solid.
m/z ES+ [M+H]+ 706.2; 1H NMR (400 MHz, CDCl3) δ ppm 8.68 (d, J=17.2 Hz, 1H), 8.32 (s, 1H), 7.51-7.45 (m, 1H), 7.44-7.37 (m, 4H), 7.37-7.30 (m, 3H), 7.06 (s, 1H), 5.24 (d, J=1.6 Hz, 2H), 4.82-4.73 (m, 4H), 4.69-4.66 (m, 1H), 4.43-4.28 (m, 2H), 3.60-3.56 (m, 1H), 3.40-3.29 (m, 3H), 3.15-3.05 (m, 1H), 3.03-2.89 (m, 1H), 2.73-2.66 (m, 1H), 2.48 (s, 3H), 2.25-2.17 (m, 1H).
Step b. A mixture of benzyl 2-(((3aR,11aS)-6-chloro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)methyl)-5,7-dihydro-6H-pyrrolo[3,4-b]pyrazine-6-carboxylate (2.66 g, 3.77 mmol), methylboronic acid (4.51 g, 75.3 mmol), Cy3P—Pd-G3 (244 mg, 0.38 mmol) and Cs2CO3 (3.68 g, 11.3 mmol) in toluene (100 mL) was degassed and purged with N2 3 times. The mixture was stirred at 100° C. for 16 h under a N2 atmosphere. Upon completion, the reaction mixture was diluted with H2O (150 mL) and extracted with EtOAc (3×150 mL). The combined organic layers were washed with brine (400 mL), dried over Na2SO4 and evaporated. The residue was purified by column chromatography (PE/EtOAc=1/3) followed by reverse phase flash chromatography (water (0.1% NH4OH)/MeCN) to give benzyl 2-(((3aR,11aS)-6,10-dimethyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)methyl)-5,7-dihydro-6H-pyrrolo[3,4-b]pyrazine-6-carboxylate (2 g, 75% yield) as a yellow solid.
m/z ES+ [M+H]+ 686.3; 1H NMR (400 MHz, CDCl3) δ ppm 8.49 (s, 1H), 8.33 (s, 1H), 7.45-7.32 (m, 5H), 7.25 (s, 3H), 7.05 (s, 1H), 5.25 (s, 2H), 4.84-4.73 (m, 5H), 4.34-4.33 (m, 1H), 4.22-4.15 (m, 1H), 3.55-3.54 (m, 1H), 3.40 (d, J=16.4 Hz, 3H), 3.15-3.01 (m, 1H), 2.91-2.81 (m, 1H), 2.76-2.67 (m, 1H), 2.50 (s, 3H), 2.41 (s, 3H), 2.21-2.14 (m, 1H).
Step c. A mixture of benzyl 2-(((3aR,11aS)-6,10-dimethyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)methyl)-5,7-dihydro-6H-pyrrolo[3,4-b]pyrazine-6-carboxylate (2.0 g, 2.92 mmol) and 10% Pd/C (50% w/w H2O; 400 mg) in 2-propanol (40 mL) was degassed and purged with H2 (15 psi) 3 times. The mixture was stirred at 45° C. under a H2 atmosphere (15 psi) for 3 h. Upon completion, the reaction mixture was filtered and evaporated to give (3aR,11aS)-5-((6,7-dihydro-5H-pyrrolo[3,4-b]pyrazin-2-yl)methyl)-6,10-dimethyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (1.61 g, crude) as a yellow solid.
m/z ES+ [M+H]+ 552.2; 1H NMR (400 MHz, CDCl3) δ ppm 8.41 (s, 1H), 8.33 (s, 1H), 7.26-7.22 (m, 3H), 7.05 (s, 1H), 4.76 (d, J=9.6 Hz, 1H), 4.40-4.29 (m, 5H), 4.16 (d, J=14.8 Hz, 1H), 4.07-3.99 (m, 1H), 3.59 (dd, J=14.0, 4.4 Hz, 1H), 3.40 (s, 3H), 3.16-3.04 (m, 1H), 2.84-2.74 (m, 1H), 2.72-2.68 (m, 1H), 2.50 (s, 3H), 2.42 (s, 3H), 2.22-2.18 (m, 1H).
Step d. Acryloyl chloride (479 mg, 5.30 mmol) was added dropwise to a solution of (3aR,11aS)-5-((6,7-dihydro-5H-pyrrolo[3,4-b]pyrazin-2-yl)methyl)-6,10-dimethyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (1.95 g, 3.54 mmol) and DIPEA (913 mg, 7.07 mmol) in DCM (30 mL) at 0° C. The reaction mixture was stirred at 0° C. for 10 min. Upon completion, the reaction mixture was evaporated and purified by column chromatography (EtOAc) followed by reverse phase flash chromatography (water (0.1% NH4OH)/MeCN) to give the title compound (1.81 g, 85% yield) as a white solid.
m/z ES+ [M+H]+ 606.2; 1H NMR (400 MHz, CDCl3) δ ppm 8.53 (d, J=3.2 Hz, 1H), 8.33 (s, 1H), 7.27-7.21 (m, 3H), 7.05 (s, 1H), 6.61-6.49 (m, 2H), 5.88-5.82 (m, 1H), 4.97 (s, 2H), 4.91 (d, J=8.4 Hz, 2H), 4.77-4.74 (m, 1H), 4.39-4.34 (m, 1H), 4.25-4.17 (m, 1H), 3.61-2.57 (m, 1H), 3.46-3.33 (m, 3H), 3.19-3.01 (m, 1H), 2.93-2.83 (m, 1H), 2.77-2.68 (m, 1H), 2.50 (s, 3H), 2.42 (d, J=6.0 Hz, 3H), 2.22-2.15 (m, 1H).
Step a. This step was conducted as described in Example 228, step a.
Step b. A solution of benzyl 2-(((3aR,11aS)-6-chloro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)methyl)-5,7-dihydro-6H-pyrrolo[3,4-b]pyrazine-6-carboxylate (150 mg, 0.21 mmol) in TFA (2 mL) was stirred at 60° C. for 3 h. Upon completion, the reaction mixture was evaporated. The residue was dissolved in DCM (3 mL), basified to pH 8 with sat. aq. Na2CO3 and extracted with EtOAc (3×8 mL). The combined organic layers were washed with brine (15 mL), dried over Na2SO4 and evaporated to give (3aR,11aS)-6-chloro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-5-((6-(2,2,2-trifluoroacetyl)-6,7-dihydro-5H-pyrrolo[3,4-b]pyrazin-2-yl)methyl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (150 mg, crude) as a black solid.
m/z ES+ [M+H]+ 668.1.
Step c. To a solution of (3aR,11aS)-6-chloro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-5-((6-(2,2,2-trifluoroacetyl)-6,7-dihydro-5H-pyrrolo[3,4-b]pyrazin-2-yl)methyl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (150 mg, 0.22 mmol) in MeOH (4 mL) and H2O (1 mL) was added Na2CO3 (48 mg, 0.45 mmol). The mixture was stirred at 20° C. for 1 h. Upon completion, the reaction mixture was diluted with H2O (8 mL) and extracted with EtOAc (3×10 mL). The combined layers were washed with brine (20 mL), dried over Na2SO4 and evaporated to give (3aR,11aS)-6-chloro-5-((6,7-dihydro-5H-pyrrolo[3,4-b]pyrazin-2-yl)methyl)-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (115 mg, crude) as a black solid.
m/z ES+ [M+H]+ 572.0.
Step d. This step was conducted in a similar manner to Example 228, step c.
m/z ES+ [M+H]+ 626.0; 1H NMR (400 MHz, CDCl3) δ ppm 8.75-8.65 (m, 1H), 8.32 (s, 1H), 7.52-7.46 (m, 1H), 7.36-7.30 (m, 2H), 7.06 (s, 1H), 6.61-6.49 (m, 2H), 5.88-5.80 (m, 1H), 4.97 (s, 2H), 4.91 (d, J=9.2 Hz, 2H), 4.70-4.66 (m, 1H), 4.44-4.31 (m, 2H), 3.62-3.57 (m, 1H), 3.42-3.25 (m, 3H), 3.16-3.07 (m, 1H), 3.06-2.91 (m, 1H), 2.73-2.67 (m, 1H), 2.48 (s, 3H), 2.25-2.18 (m, 1H).
Step a. To a solution of Intermediate 5b (615 mg, 1.40 mmol), Intermediate B51a (550 mg, 1.40 mmol) in DMF (6 mL) was added TMSCI (381 mg, 3.50 mmol). The reaction mixture was stirred at 0° C. for 30 min. A solution of borane tetrahydrofuran complex (1 M in THF, 2.1 mL) in THF (6 mL) was added and the reaction mixture was stirred at 30° C. for 12 h. Upon completion, the reaction mixture was quenched with sat. aq. NaHCO3 (10 mL), diluted with water (70 mL) and extracted with EtOAc (3×30 mL). The combined organic layers were washed with brine (3×30 mL), dried over Na2SO4 and evaporated. The residue was purified by column chromatography (PE/EtOAc=1/3) to give tert-butyl (R/S)-3-(((3aR,11aS)-6-chloro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)methyl)-7-(hydroxymethyl)-5,7-dihydro-6H-pyrrolo[3,4-b]pyridine-6-carboxylate (530 mg, 54% yield) as a yellow solid.
m/z ES+ [M+H]+ 701.3.
Step b. To a solution of tert-butyl (R/S)-3-(((3aR,11aS)-6-chloro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)methyl)-7-(hydroxymethyl)-5,7-dihydro-6H-pyrrolo[3,4-b]pyridine-6-carboxylate (480 mg, 0.69 mmol) in DCM (4 mL) was added TFA (1.54 g, 13.5 mmol). The reaction mixture was stirred for 1 h at 25° C. Upon completion, the reaction mixture was evaporated to give (3aR,11aS)-6-chloro-5-(((R/S)-7-(hydroxymethyl)-6,7-dihydro-5H-pyrrolo[3,4-b]pyridin-3-yl)methyl)-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (411 mg, crude) as a yellow oil.
m/z ES+ [M+H]+ 601.2.
Step c. To a solution of (3aR,11aS)-6-chloro-5-(((R/S)-7-(hydroxymethyl)-6,7-dihydro-5H-pyrrolo[3,4-b]pyridin-3-yl)methyl)-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (400 mg, 0.67 mmol) and DIPEA (258 mg, 2.00 mmol) in DCM (5 mL) was added acryloyl chloride (57 mg, 0.63 mmol) at −40° C. The reaction mixture was stirred at −40° C. for 5 min. Upon completion, the mixture was quenched with water (1 mL), further diluted with water (30 mL) and extracted with DCM (2×30 mL). The combined organic layers were washed with brine (30 mL), dried over Na2SO4 and evaporated. The residue was purified by reverse phase flash chromatography (water (0.1% NH4OH)/MeCN) to give ((R/S)-6-acryloyl-3-(((3aR,11aS)-6-chloro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)methyl)-6,7-dihydro-5H-pyrrolo[3,4-b]pyridin-7-yl)methyl acrylate (250 mg, 53% yield) as a white solid.
m/z ES+ [M+H]+ 709.3.
Step d. To a solution of ((R/S)-6-acryloyl-3-(((3aR,11aS)-6-chloro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)methyl)-6,7-dihydro-5H-pyrrolo[3,4-b]pyridin-7-yl)methyl acrylate (230 mg, 0.32 mmol) in THF (1 mL) and H2O (0.5 mL) was added lithium hydroxide monohydrate (41 mg, 0.97 mmol). The reaction mixture was stirred at 30° C. for 1 h. Upon completion, the mixture was quenched with water (1 mL), further diluted with water (30 mL) and extracted with EtOAc (2×30 mL). The combined organic layers were washed with brine (30 mL), dried over Na2SO4 and evaporated. The residue was purified by reverse phase flash chromatography (water (0.1% NH4OH)/MeCN) to give the title compound (120 mg, 56% yield) as a white solid.
m/z ES+ [M+H]+ 655.2; 1H NMR (400 MHz, CDCl3) δ ppm 8.45 (s, 1H), 8.33 (s, 1H), 7.85-7.68 (m, 1H), 7.54-7.45 (m, 1H), 7.35-7.30 (m, 2H), 7.07 (s, 1H), 6.64-6.48 (m, 2H), 5.91-5.82 (m, 1H), 5.45-5.36 (m, 1H), 4.96 (d, J=3.2 Hz, 2H), 4.66 (d, J=9.6 Hz, 1H), 4.33-4.16 (m, 3H), 4.00-3.83 (m, 1H), 3.52-3.46 (m, 1H), 3.34-3.28 (m, 3H), 3.11-3.01 (m, 1H), 2.97-2.81 (m, 1H), 2.73-2.63 (m, 1H), 2.49 (s, 3H), 2.25-2.16 (m, 1H).
The title compound was prepared in a similar manner to Example 230, using Intermediate B51b in step a.
m/z ES+ [M+H]+ 655.2; 1H NMR (400 MHz, CDCl3) δ ppm 8.53-8.44 (m, 1H), 8.32 (s, 1H), 7.79-7.67 (m, 1H), 7.50-7.47 (m 1H), 7.37-7.28 (m, 2H), 7.06 (s, 1H), 6.71-6.46 (m, 2H), 5.91-5.77 (m, 1H), 5.43-5.11 (m, 1H), 5.08-4.90 (m, 2H), 4.78-4.62 (m, 1H), 4.34-4.21 (m, 2H), 4.21-4.05 (m, 1H), 4.00-3.84 (m, 1H), 3.50-3.47 (m, 1H), 3.39-3.19 (m, 3H), 3.10-2.98 (m, 1H), 2.97-2.84 (m, 1H), 2.69-2.64 (m, 1H), 2.48 (s, 3H), 2.24-2.16 (m, 1H).
Step a. This step was conducted as described in Example 230.
Step b. This step was conducted in a similar manner as Example 77, step b.
m/z ES+ [M+H]+ 681.4; 1H NMR (400 MHz, CDCl3) δ ppm 8.43 (s, 1H), 8.33 (s, 1H), 7.66-7.51 (m, 1H), 7.27 (s, 3H), 7.06 (s, 1H), 5.19-4.89 (m, 1H), 4.79-4.40 (m, 4H), 4.18 (d, J=14.4 Hz, 2H), 4.09-3.87 (m, 2H), 3.56-3.23 (m, 4H), 3.05-2.61 (m, 3H), 2.54-2.47 (m, 3H), 2.45 (s, 3H), 2.22-2.07 (m, 1H), 1.54 (s, 9H).
Steps c-d. These 2 steps were conducted in a similar manner to Example 27, steps b-c, using acrylic acid in step d.
m/z ES+ [M+H]+ 635.5; 1H NMR (400 MHz, CDCl3) δ ppm 8.56-8.49 (m, 1H), 8.32 (s, 1H), 7.89-7.64 (m, 1H), 7.31-7.27 (m, 3H), 7.07 (s, 1H), 6.72-6.48 (m, 2H), 5.93-5.80 (m, 1H), 5.53-5.30 (m, 1H), 5.17-4.78 (m, 2H), 4.78-4.70 (m, 1H), 4.36-4.27 (m, 1H), 4.26-4.17 (m, 1H), 4.17-4.06 (m, 1H), 4.06-3.86 (m, 1H), 3.45-3.50 (m, 1H), 3.42-3.34 (m, 3H), 3.07-2.87 (m, 1H), 2.87-2.78 (m, 1H), 2.60-2.75 (m, 1H), 2.51 (s, 3H), 2.47 (s, 3H), 2.21-2.12 (m, 1H).
The title compound was prepared in a similar manner to Example 232, using Intermediate B51b in step a.
m/z ES+ [M+H]+ 635.4; 1H NMR (400 MHz, CDCl3) δ ppm 8.57-8.46 (m, 1H), 8.33 (s, 1H), 7.69-7.56 (m, 1H), 7.31-7.27 (m, 2H), 7.25 (s, 1H), 7.07 (s, 1H), 6.72-6.49 (m, 2H), 5.92-5.81 (m, 1H), 5.46-5.13 (m, 1H), 5.11-4.77 (m, 2H), 4.76-4.70 (m, 1H), 4.36-4.27 (m, 1H), 4.27-4.19 (m, 1H), 4.17-4.03 (m, 1H), 4.03-3.85 (m, 1H), 3.49-3.41 (m, 1H), 3.42-3.32 (m, 3H), 3.08-2.89 (m, 1H), 2.88-2.75 (m, 1H), 2.74-2.63 (m, 1H), 2.51 (s, 3H), 2.49-2.43 (m, 3H), 2.23-2.12 (m, 1H).
Step a. To a solution of Intermediate B51b (215 mg, 0.55 mmol) and Intermediate 5b (0.2 g, 0.46 mmol) in DMF (5 mL) was added TMSCI (124 mg, 1.14 mmol) at 0° C. The mixture was stirred for 30 min, after which a solution of borane tetrahydrofuran complex (1 M in THF, 0.46 mL) in THF (5 mL) was added. The reaction mixture was stirred at 20° C. for 11 h 30 min. Upon completion, the reaction mixture was quenched with sat. aq. NaHCO3 solution (30 mL) and extracted with EtOAc (30 mL). The organic layer was washed with brine (30 mL), dried over Na2SO4, evaporated and purified by column chromatography (EtOAc) to give tert-butyl (S/R)-3-(((3aR,11aS)-6-chloro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)methyl)-7-(hydroxymethyl)-5,7-dihydro-6H-pyrrolo[3,4-b]pyridine-6-carboxylate (220 mg, 64% yield) as a colourless gum.
m/z ES+ [M+H]+ 701.5, 1H NMR (400 MHz, CDCl3) δ ppm 8.52-8.38 (m, 1H), 8.33 (s, 1H), 7.71-7.63 (m, 1H), 7.52-7.45 (m, 1H), 7.32 (d, J=4.0 Hz, 2H), 7.06 (s, 1H), 5.16-4.87 (m, 1H), 4.84-4.56 (m, 3H), 4.34-4.25 (m, 1H), 4.24-4.16 (m, 3H), 3.98-3.86 (m, 1H), 3.53-3.44 (m, 1H), 3.36-3.24 (m, 3H), 3.09-2.99 (m, 1H), 2.98-2.85 (m, 1H), 2.74-2.63 (m, 1H), 2.48 (s, 3H), 2.26-2.15 (m, 1H), 1.53 (s, 9H).
Step b. A mixture of tert-butyl (S/R)-3-(((3aR,11aS)-6-chloro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)methyl)-7-(hydroxymethyl)-5,7-dihydro-6H-pyrrolo[3,4-b]pyridine-6-carboxylate (0.21 g, 0.30 mmol), methylboronic acid (717 mg, 12.0 mmol), Cy3P—Pd-G3 (39 mg, 0.06 mmol) and Cs2CO3 (293 mg, 0.90 mmol) in 1,4-dioxane (6 mL) was stirred at 100° C. for 12 h under a N2 atmosphere. Upon completion, the reaction mixture was evaporated, the residue was diluted with water (20 mL) and extracted with EtOAc (20 mL). The organic layer was washed with brine (20 mL), dried over Na2SO4, evaporated and purified by column chromatography (EtOAc) to give tert-butyl (S/R)-3-(((3aR,11aS)-6,10-dimethyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)methyl)-7-(hydroxymethyl)-5,7-dihydro-6H-pyrrolo[3,4-b]pyridine-6-carboxylate (160 mg, 78% yield) as a white solid.
m/z ES+ [M+H]+ 681.3; 1H NMR (400 MHz, CDCl3) δ ppm 8.46 (s, 1H), 8.33 (s, 1H), 7.62-7.52 (m, 1H), 7.30-7.27 (m, 1H), 7.27-7.22 (m, 2H), 7.06 (s, 1H), 5.16-4.89 (m, 1H), 4.88-4.57 (m, 3H), 4.26-4.16 (m, 2H), 4.09-3.87 (m, 2H), 3.49-3.42 (m, 2H), 3.42-3.34 (m, 3H), 3.07-2.90 (m, 1H), 2.86-2.75 (m, 1H), 2.74-2.62 (m, 1H), 2.50 (s, 3H), 2.46 (s, 3H), 2.22-2.12 (m, 1H), 1.54 (s, 9H).
Step c. To a solution of tert-butyl (S/R)-3-(((3aR,11aS)-6,10-dimethyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)methyl)-7-(hydroxymethyl)-5,7-dihydro-6H-pyrrolo[3,4-b]pyridine-6-carboxylate (0.19 g, 0.28 mmol) in DMF (3 mL) was added NaH (13 mg, 0.34 mmol, 60% dispersion in mineral oil) at 0° C. The mixture was stirred for 30 min, after which MeI (59 mg, 0.42 mmol) was added. The reaction mixture was stirred at 0° C. for 1 h. Upon completion, the reaction mixture was poured into sat. aq. NH4Cl (20 mL) and extracted with EtOAc (20 mL). The organic layer was washed with brine (20 mL), dried over Na2SO4, evaporated and purified by prep-TLC (EtOAc) to give tert-butyl (S/R)-3-(((3aR,11aS)-6,10-dimethyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)methyl)-7-(methoxymethyl)-5,7-dihydro-6H-pyrrolo[3,4-b]pyridine-6-carboxylate (40 mg, 19% yield) as a white solid.
m/z ES+ [M+H]+ 695.3.
Step d. To a solution of tert-butyl (S/R)-3-(((3aR,11aS)-6,10-dimethyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)methyl)-7-(methoxymethyl)-5,7-dihydro-6H-pyrrolo[3,4-b]pyridine-6-carboxylate (40 mg, 0.58 mmol) in DCM (3 mL) was added TFA (1.54 g, 13.5 mmol). The reaction mixture was stirred at 25° C. for 30 min. Upon completion, the mixture was evaporated to give (3aR,11aS)-5-(((S/R)-7-(methoxymethyl)-6,7-dihydro-5H-pyrrolo[3,4-b]pyridin-3-yl)methyl)-6,10-dimethyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione as a TFA salt (40 mg, 98% yield) as a yellow gum.
m/z ES+ [M+H]+ 595.2.
Step e. To a solution of (3aR,11aS)-5-(((S/R)-7-(methoxymethyl)-6,7-dihydro-5H-pyrrolo[3,4-b]pyridin-3-yl)methyl)-6,10-dimethyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (TFA salt, 40 mg, 0.057 mmol) in DCM (5 mL) was added DIPEA (37 mg, 0.28 mmol) and acryloyl chloride (6 mg, 68 mmol). The reaction mixture was stirred at 20° C. for 10 min. Upon completion, the reaction mixture was diluted with water (20 mL) and extracted with EtOAc (20 mL). The organic layer was washed with brine (20 mL), dried over Na2SO4, evaporated and purified by prep-TLC (EtOAc) to give the title compound (36 mg, 96% yield) as a white solid.
m/z ES+ [M+H]+ 649.3, 1H NMR (400 MHz, CDCl3) δ ppm 8.48 (d, J=4.0 Hz, 1H), 8.33 (s, 1H), 7.58 (d, J=18.0 Hz, 1H), 7.28 (s, 1H), 7.27-7.21 (m, 2H), 7.06 (s, 1H), 6.77-6.54 (m, 1H), 6.54-6.45 (m, 1H), 5.84-5.75 (m, 1H), 5.40-5.20 (m, 1H), 5.13-4.64 (m, 3H), 4.30-4.04 (m, 2H), 4.03-3.77 (m, 2H), 3.52-3.42 (m, 1H), 3.41-3.29 (m, 3H), 3.28-3.17 (m, 3H), 3.08-2.93 (m, 1H), 2.86-2.74 (m, 1H), 2.73-2.64 (m, 1H), 2.50 (s, 3H), 2.45 (d, J=7.2 Hz, 3H), 2.22-2.11 (m, 1H).
The title compound was prepared in a similar manner to Example 234, using Intermediate B51a in step a.
m/z ES+ [M+H]+ 649.2, 1H NMR (400 MHz, CDCl3) δ ppm 8.52-8.41 (m, 1H), 8.32 (s, 1H), 7.67-7.56 (m, 1H), 7.31-7.28 (m, 1H), 7.25 (s, 2H), 7.06 (s, 1H), 6.76-6.54 (m, 1H), 6.54-6.44 (m, 1H), 5.88-5.72 (m, 1H), 5.43-5.19 (m, 1H), 5.10-4.68 (m, 3H), 4.29-4.10 (m, 2H), 4.09-3.79 (m, 2H), 3.48 (d, J=13.2 Hz, 1H), 3.42-3.30 (m, 3H), 3.28-3.16 (m, 3H), 3.07-2.90 (m, 1H), 2.88-2.75 (m, 1H), 2.74-2.63 (m, 1H), 2.50 (s, 3H), 2.46 (d, J=7.2 Hz, 3H), 2.23-2.12 (m, 1H).
Steps a-d. These 4 steps were conducted in a similar manner to Example 234, steps a, c-e, using Intermediate B51b in step a.
m/z ES+ [M+H]+ 669.2; 1H NMR (400 MHz, CDCl3) δ ppm 8.53-8.43 (m, 1H), 8.33 (s, 1H), 7.72-7.63 (m, 1H), 7.52-7.45 (m, 1H), 7.37-7.28 (m, 2H), 7.06 (s, 1H), 6.75-6.44 (m, 2H), 5.83-5.74 (m, 1H), 5.37-5.19 (m, 1H), 5.11-4.63 (m, 3H), 4.37-4.28 (m, 1H), 3.96-3.90 (m, 1H), 4.28-3.76 (m, 2H), 3.57-3.43 (m, 1H), 3.35-3.16 (m, 6H), 3.11-2.86 (m, 2H), 2.76-2.63 (m, 1H), 2.48 (s, 3H), 2.27-2.14 (m, 1H).
The title compound was prepared in a similar manner to Example 236, using Intermediate B51a in step a.
m/z ES+ [M+H]+ 669.2, 1H NMR (400 MHz, CDCl3) δ ppm 8.49-8.37 (m, 1H), 8.32 (s, 1H), 7.77-7.64 (m, 1H), 7.48 (d, J=6.8 Hz, 1H), 7.36-7.28 (m, 2H), 7.12-7.01 (m, 1H), 6.76-6.36 (m, 2H), 5.85-5.73 (m, 1H), 5.38-5.17 (m, 1H), 5.10-4.79 (m, 2H), 4.73-4.60 (m, 1H), 4.35-4.12 (m, 2H), 3.96-3.76 (m, 2H), 3.58-3.46 (m, 1H), 3.35-3.15 (m, 6H), 3.11-2.85 (m, 2H), 2.75-2.63 (m, 1H), 2.48 (s, 3H), 2.27-2.20 (m, 1H).
Step a. This step was conducted in a similar manner to Example 228, step a, using Intermediate B52.
m/z ES+ [M+H]+ 939.4; 1H NMR (400 MHz, CDCl3) δ ppm 8.33 (s, 1H), 7.66-7.56 (m, 2H), 7.56-7.45 (m, 4H), 7.44-7.29 (m, 9H), 7.06 (s, 1H), 5.17-5.00 (m, 1H), 4.84-4.62 (m, 3H), 4.51-4.22 (m, 3H), 4.00-3.89 (m, 1H), 3.57-3.49 (m, 1H), 3.46-3.36 (m, 3H), 3.18-2.93 (m, 2H), 2.70-2.55 (m, 1H), 2.49 (s, 3H), 2.24-2.13 (m, 1H), 1.56-1.39 (m, 9H), 0.90-0.81 (m, 9H).
Steps b-d. These 3 steps were conducted in a similar manner to Example 232, steps b-d.
m/z ES+ [M+H]+ 873.1; 1H NMR (400 MHz, CDCl3) δ ppm 8.33 (s, 1H), 7.67-7.53 (m, 2H), 7.49-7.42 (m, 3H), 7.42-7.35 (m, 5H), 7.34-7.29 (m, 3H), 7.26-7.24 (m, 2H), 7.06 (s, 1H), 6.63-6.38 (m, 2H), 5.83-5.80 (m, 1H), 5.50 (s, 1H), 4.95-4.89 (m, 1H), 4.80-4.75 (m, 1H), 4.42-4.17 (m, 3H), 4.02-3.97 (m, 1H), 3.57-3.47 (m, 1H), 3.46-3.39 (m, 3H), 3.18-3.03 (m, 1H), 2.90-2.78 (m, 1H), 2.72-2.58 (m, 1H), 2.51 (s, 3H), 2.49-2.47 (m, 3H), 2.27-2.07 (m, 2H), 0.97-0.86 (m, 9H).
Step e. To a solution of (3aR,11aS)-5-((6-acryloyl-5-(((tert-butyldiphenylsilyl)oxy)methyl)-6,7-dihydro-5H-pyrrolo[3,4-b]pyridin-2-yl)methyl)-6,10-dimethyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (20 mg, 0.023 mmol) in THF (0.5 mL) was added TBAF (1 M in THF, 23 μL). The mixture was stirred at 20° C. for 3 h. Upon completion, the reaction mixture was evaporated and purified by Prep-TLC (EtOAc/MeOH=10/1) to give the title compound (11 mg, 73% yield) as an off-white solid.
m/z ES+ [M+H]+ 635.3; 1H NMR (400 MHz, CDCl3) δ ppm 8.32 (s, 1H), 7.68-7.63 (m, 1H), 7.40 (t, J=8.4 Hz, 1H), 7.27-7.22 (m, 3H), 7.05 (s, 1H), 6.66-6.50 (m, 2H), 5.91-5.88 (m, 1H), 5.54 (s, 1H), 4.93 (d, J=4.8 Hz, 2H), 4.87-4.71 (m, 2H), 4.38-4.31 (m, 1H), 4.23-4.15 (m, 1H), 4.06-3.98 (m, 1H), 3.92-3.83 (m, 1H), 3.55-3.50 (m, 1H), 3.36 (d, J=8.8 Hz, 3H), 3.15-3.03 (m, 1H), 2.89-2.85 (m, 1H), 2.73-2.64 (m, 1H), 2.50 (s, 3H), 2.47 (s, 3H), 2.21-2.12 (m, 1H).
Step a. This step was conducted as described in Example 238, step a.
Steps b-d. These 3 steps were conducted in a similar manner to Example 238, steps c-e.
m/z ES+ [M+H]+ 655.0; 1H NMR (400 MHz, CDCl3) δ ppm 8.32 (s, 1H), 7.68-7.63 (m, 1H), 7.61-7.55 (m, 1H), 7.51-7.47 (m, 1H), 7.35-7.30 (m, 2H), 7.06 (s, 1H), 6.65-6.50 (m, 2H), 5.90-5.87 (m, 1H), 5.54 (s, 1H), 4.93 (d, J=4.8 Hz, 2H), 4.79-4.65 (m, 2H), 4.35 (s, 2H), 4.05-3.97 (m, 1H), 3.91-3.81 (m, 1H), 3.56-3.53 (m, 1H), 3.31 (d, J=12.4 Hz, 3H), 3.15-2.98 (m, 2H), 2.73-2.63 (m, 1H), 2.48 (s, 3H), 2.25-2.15 (m, 1H).
Step a. To a solution of Intermediate 5b (45.0 g, 102 mmol) and Intermediate B53 (26.8 g, 102 mmol) in DMF (450 mL) was added TMSCI (27.8 g, 256 mmol) at 0° C. The mixture was stirred at 0° C. for 30 min. A solution of borane tetrahydrofuran complex (1 M in THF, 102 mL) was then added dropwise at 0° C. The reaction mixture was stirred at 25° C. for 3 h. Upon completion, the reaction mixture was quenched with MeOH (350 mL) and sat. aq. NaHCO3 (1000 mL) was added. The aqueous mixture was extracted with EtOAc (3×1000 mL). The combined organic layers were washed with brine (1000 mL), dried over Na2SO4 and evaporated. The residue was purified by column chromatography (PE/EtOAc=10/1 to 2/1) to give tert-butyl 2-(((3aR,11aS)-6-chloro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)methyl)-5,7-dihydro-6H-pyrrolo[3,4-b]pyridine-6-carboxylate (52.8 g, 72% yield) as a yellow solid.
m/z ES+ [M+H]+ 671.4; 1H NMR (400 MHz, CDCl3) δ ppm 8.32 (s, 1H), 7.63-7.52 (m, 2H), 7.50-7.46 (m, 1H), 7.35-7.30 (m, 2H), 7.05 (s, 1H), 4.73-4.61 (m, 5H), 4.46-4.35 (m, 1H), 3.59-3.50 (m, 1H), 3.48-3.41 (m, 2H), 3.37-3.34 (m, 2H), 3.13-3.04 (m, 1H), 3.02-2.89 (m, 1H), 2.72-2.63 (m, 1H), 2.48 (s, 3H), 2.24-2.14 (m, 1H), 1.52 (s, 9H).
Step b. To a solution of tert-butyl 2-(((3aR,11aS)-6-chloro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)methyl)-5,7-dihydro-6H-pyrrolo[3,4-b]pyridine-6-carboxylate (52.8 g, 73.9 mmol) and Cy3P—Pd-G3 (2.80 g, 3.81 mmol) in 1,4-dioxane (510 mL) was added Cs2CO3 (74.5 g, 228 mmol) at 25° C. Methylboronic acid (27.3 g, 457 mmol) in 1,4-dioxane (150 mL) was then added dropwise at 90° C. The resulting mixture was stirred at 110° C. for 4 h. Upon completion, the reaction mixture was diluted with water (1000 mL) and extracted with EtOAc (3×500 mL). The combined organic layers were dried over Na2SO4 and evaporated. The residue was purified by column chromatography (PE/EtOAc=7/1 to 2/1) to give tert-butyl 2-(((3aR,11aS)-6,10-dimethyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)methyl)-5,7-dihydro-6H-pyrrolo[3,4-b]pyridine-6-carboxylate (44.0 g, 67% yield) as a white solid.
m/z ES+ [M+H]+ 651.4; 1H NMR (400 MHz, CDCl3) δ ppm 8.32 (s, 1H), 7.56 (dd, J=16.4, 7.6 Hz, 1H), 7.32 (dd, J=7.6, 3.2 Hz, 1H), 7.25 (s, 3H), 7.05 (s, 1H), 4.79-4.73 (m, 1H), 4.71-4.60 (m, 4H), 4.35-4.24 (m, 1H), 3.54 (td, J=14.0, 4.0 Hz, 1H), 3.44-3.37 (m, 4H), 3.12-2.95 (m, 1H), 2.92-2.78 (m, 1H), 2.72-2.60 (m, 1H), 2.50 (s, 3H), 2.46 (s, 3H), 2.16 (dd, J=16.8, 11.6 Hz, 1H), 1.53 (s, 9H).
Step c. To a solution of tert-butyl 2-(((3aR,11aS)-6,10-dimethyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)methyl)-5,7-dihydro-6H-pyrrolo[3,4-b]pyridine-6-carboxylate (36.5 g, 41.5 mmol) in DCM (365 mL) was added TFA (54 mL, 730 mmol). The reaction mixture was stirred at 25° C. for 30 min. Upon completion, the reaction mixture was evaporated to give (3aR,11aS)-5-((6,7-dihydro-5H-pyrrolo[3,4-b]pyridin-2-yl)methyl)-6,10-dimethyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione as a TFA salt (40.0 g, crude) as a brown oil that was used without further purification.
m/z ES+ [M+H]+ 551.2.
Step d. DIPEA was added to a solution of (3aR,11aS)-5-((6,7-dihydro-5H-pyrrolo[3,4-b]pyridin-2-yl)methyl)-6,10-dimethyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (35.7 g, 53.7 mmol) in DCM (360 mL) until the pH of the mixture was ˜7. After which, additional DIPEA (18.7 mL, 107 mmol) was added followed by the dropwise addition of acryloyl chloride (6.79 mL, 83.2 mmol) in DCM (36 mL) at 0° C. The resulting mixture was stirred at 0° C. for 10 min. Upon completion, the reaction mixture was diluted with water (1000 mL) and extracted with EtOAc (3×500 mL). The combined organic layers were dried over Na2SO4, filtered and evaporated. The residue was purified by reverse phase flash chromatography (water (0.1% FA)/MeCN) to give the title compound (20.0 g, 60% yield) as a white solid.
m/z ES+ [M+H]+ 605.2; 1H NMR (400 MHz, CDCl3) δ ppm 8.32 (s, 1H), 7.67-7.57 (m, 1H), 7.36 (d, J=7.6 Hz, 1H), 7.27-7.23 (m, 3H), 7.05 (s, 1H), 6.62-6.47 (m, 2H), 5.83-5.78 (m, 1H), 4.95-4.86 (m, 4H), 4.76 (d, J=9.6 Hz, 1H), 4.36-4.16 (m, 2H), 3.57-3.51 (m, 1H), 3.42-3.39 (m, 3H), 3.15-2.96 (m, 1H), 2.90-2.81 (m, 1H), 2.72-2.63 (m, 1H), 2.50 (s, 3H), 2.47 (s, 3H), 2.19-2.12 (m, 1H).
Step a. To a solution of Intermediate B63 (500 mg, 1.99 mmol) and Intermediate 5b (873 mg, 1.99 mmol) in DMF (50 mL) was added TMSCI (540 mg, 4.97 mmol). The mixture was stirred at 0° C. for 30 min. A solution of borane tetrahydrofuran complex (1 M in THF, 1.99 mL) in THF (40 mL) was added. The mixture was stirred at 25° C. for 12 h 30 min. Upon completion, the reaction mixture was evaporated and purified by reverse phase flash chromatography (water (0.1% FA)/MeCN) to give tert-butyl 2-(((3aR,11aS)-6-chloro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)methyl)-1-methyl-4,6-dihydropyrrolo[3,4-d]imidazole-5(1H)-carboxylate (600 mg, 36% yield) as a yellow solid.
m/z ES+ [M+H]+ 674.4; 1H NMR (400 MHz, CDCl3) δ ppm 8.34 (s, 1H), 7.51-7.44 (m, 1H), 7.37-7.32 (m, 2H), 7.06 (s, 1H), 4.64-4.47 (m, 1H), 4.50-4.29 (m, 5H), 4.27-4.16 (m, 1H), 3.69-3.55 (m, 4H), 3.35 (d, J=13.2 Hz, 3H), 3.09-2.93 (m, 2H), 2.80-2.74 (m, 1H), 2.48 (s, 3H), 2.24-2.17 (m, 1H), 1.52 (s, 9H).
Step b. A mixture of tert-butyl 2-(((3aR,11aS)-6-chloro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)methyl)-1-methyl-4,6-dihydropyrrolo[3,4-d]imidazole-5(1H)-carboxylate (560 mg, 0.83 mmol), methylboronic acid (2.49 g, 41.5 mmol), XPhos-Pd-G2 (65.4 mg, 0.083 mmol) and Cs2CO3 (676 mg, 2.08 mmol) in toluene (50 mL) was degassed and purged with N2 3 times. The reaction mixture was stirred at 110° C. for 12 h under a N2 atmosphere. Upon completion, the reaction mixture was filtered, evaporated and purified by reverse phase flash chromatography (water (0.1% NH4OH)/MeCN) to give tert-butyl 2-(((3aR,11aS)-6,10-dimethyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)methyl)-1-methyl-4,6-dihydropyrrolo[3,4-d]imidazole-5(1H)-carboxylate (350 mg, 61% yield) as a colourless oil.
m/z ES+ [M+H]+ 654.3; 1H NMR (400 MHz, CDCl3) δ ppm 8.34 (s, 1H), 7.28-7.27 (m, 3H), 7.06 (s, 1H), 4.72-4.68 (m, 1H), 4.51-4.27 (m, 5H), 4.16-4.01 (m, 1H), 3.73-3.61 (m, 1H), 3.57 (d, J=1.6 Hz, 3H), 3.40 (d, J=10.4 Hz, 3H), 3.19-2.91 (m, 1H), 2.84-2.72 (m, 2H), 2.50 (s, 3H), 2.41-2.40 (m, 3H), 2.21-2.18 (m, 1H), 1.52 (s, 9H).
Step c. A mixture of tert-butyl 2-(((3aR,11aS)-6,10-dimethyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)methyl)-1-methyl-4,6-dihydropyrrolo[3,4-d]imidazole-5(1H)-carboxylate (300 mg, 0.46 mmol) and TFA (523 mg, 4.6 mmol) in DCM (1 mL) was stirred at 25° C. for 1 h under a N2 atmosphere. Upon completion, the reaction mixture was evaporated to give (3aR,11aS)-6,10-dimethyl-5-((1-methyl-1,4,5,6-tetrahydropyrrolo[3,4-d]imidazol-2-yl)methyl)-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione as the TFA salt (300 mg, crude) as a yellow oil.
m/z ES+ [M+H]+ 554.3.
Step d. To a solution of (3aR,11aS)-6,10-dimethyl-5-((1-methyl-1,4,5,6-tetrahydropyrrolo[3,4-d]imidazol-2-yl)methyl)-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (300 mg, 0.54 mmol), acrylic acid (51 mg, 0.70 mmol) in DCM (2 mL) was added DIPEA (210 mg, 1.6 mmol) and HATU (288 mg, 0.76 mmol). The mixture was stirred at 25° C. for 10 min. Upon completion, the reaction mixture was diluted with water (50 mL) and extracted with DCM (3×30 mL). The combined organic layers were washed with brine (30 mL), dried over Na2SO4 and evaporated. The residue was purified by reverse phase flash (water (0.1% NH4OH)/MeCN) to give the title compound (130 mg, 40% yield) as a white solid.
m/z ES+ [M+H]+ 608.3; 1H NMR (400 MHz, CDCl3) δ ppm 8.34 (s, 1H), 7.29-7.25 (m, 3H), 7.06 (s, 1H), 6.62-6.42 (m, 2H), 5.81 (dd, J=7.6, 4.4 Hz, 1H), 4.83-4.52 (m, 5H), 4.39-4.28 (m, 1H), 4.17-3.99 (m, 1H), 3.74-3.54 (m, 4H), 3.50-3.34 (m, 3H), 3.21-2.93 (m, 1H), 2.88-2.71 (m, 2H), 2.51 (s, 3H), 2.45-2.37 (m, 3H), 2.19 (dd, J=17.2, 11.2 Hz, 1H).
Step a. This step was conducted as described in Example 241, step a.
Steps b-c. These two steps were conducted in a similar manner to Example 241, steps c-d.
m/z ES+ [M+H]+ 628.2; 1H NMR (400 MHz, CDCl3) ppm 8.34 (s, 1H), 7.54-7.45 (m, 1H), 7.40-7.31 (m, 2H), 7.07 (s, 1H), 6.56-6.37 (m, 2H), 5.84-5.68 (m, 1H), 4.81-4.53 (m, 5H), 4.45-4.36 (m, 1H), 4.29-4.14 (m, 1H), 3.75-3.65 (m, 3H), 3.62-3.56 (m, 1H), 3.43-3.29 (m, 3H), 3.13-2.94 (m, 2H), 2.83-2.71 (m, 1H), 2.49 (s, 3H), 2.31-2.16 (m, 1H).
Step a. This step was conducted in a similar manner to Example 39, step a, using Intermediate 5b and Intermediate B69.
m/z ES+ [M+H]+ 620.1; 1H NMR (400 MHz, CDCl3) δ ppm 9.00 (s, 1H), 8.44 (s, 1H), 8.33 (s, 1H), 7.51-7.49 (m, 1H), 7.38-7.30 (m, 2H), 7.07 (s, 1H), 6.96-6.59 (m, 1H), 4.71 (d, J=9.6 Hz, 1H), 4.68-4.42 (m, 2H), 3.64-3.59 (m, 1H), 3.42 (s, 1H), 3.39 (s, 3H), 3.15-3.11 (m, 1H), 3.09-2.98 (m, 1H), 2.71-2.65 (m, 1H), 2.50 (s, 3H), 2.25-2.18 (m, 1H).
Step b. To a solution of (3aR,11aS)-6-chloro-5-((6-(difluoromethyl)imidazo[1,2-a]pyrazin-2-yl)methyl)-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (150 mg, 0.24 mmol) in EtOH (7.5 mL) was added NaBH3CN (46 mg, 0.73 mmol) and acetic acid (3 mg, 0.048 mmol). The reaction mixture was stirred at 25-40° C. for 56 h. The mixture was diluted with water (10 mL) and extracted with EtOAc (3×5 mL). The combined organic layers were dried over Na2SO4, filtered and evaporated. The residue was purified by reverse phase flash chromatography (water (0.1% FA)/MeCN) to give (3aR,11aS)-6-chloro-5-((6-(difluoromethyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-2-yl)methyl)-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (120 mg, 80% yield) as a white solid.
m/z ES+ [M+H]+ 624.3; 1H NMR (400 MHz, MeOD-d4) δ ppm 8.30 (s, 1H), 7.53-7.41 (m, 2H), 7.44-7.34 (m, 1H), 7.22 (s, 1H), 7.01 (s, 1H), 6.26-5.83 (m, 1H), 4.20-4.07 (m, 4H), 4.02-3.89 (m, 2H), 3.69-3.46 (m, 3H), 3.32 (s, 4H), 3.17-3.06 (m, 1H), 2.98-2.87 (m, 1H), 2.67-2.61 (m, 1H), 2.53 (s, 3H), 2.39-2.35 (m, 1H).
Step c. This step was conducted in a similar manner to Example 38, step c. An additional chiral SFC step afforded the title compounds.
Example 243a:
m/z ES+ [M+H]+ 678.1; 1H NMR (400 MHz, CDCl3) δ ppm 8.31 (s, 1H), 7.47-7.45 (m, 1H), 7.37-7.28 (m, 2H), 7.06 (s, 1H), 6.90 (s, 1H), 6.68-6.57 (m, 1H), 6.55-6.38 (m, 1H), 6.11-5.58 (m, 2H), 5.56-4.92 (m, 2H), 4.77-4.52 (m, 2H), 4.43-4.23 (m, 2H), 4.22-4.07 (m, 2H), 3.62-3.57 (m, 1H), 3.33 (s, 3H), 3.10-2.90 (m, 2H), 2.70-2.64 (m, 1H), 2.49 (s, 3H), 2.23-2.16 (m, 1H).
Example 243b:
m/z ES+ [M+H]+ 678.1; 1H NMR (400 MHz, CDCl3) δ ppm 8.32 (s, 1H), 7.47-7.45 (m, 1H), 7.35-7.28 (m, 2H), 7.05 (s, 1H), 6.90 (s, 1H), 6.72-6.56 (m, 1H), 6.55-6.36 (m, 1H), 6.07-5.64 (m, 2H), 5.57-4.92 (m, 2H), 4.78-4.50 (m, 2H), 4.41-4.20 (m, 2H), 4.20-4.09 (m, 2H), 3.63 (d, J=9.2 Hz, 1H), 3.31 (s, 3H), 3.09-2.91 (m, 2H), 2.6-2.65 (m, 1H), 2.48 (s, 3H), 2.23-2.16 (m, 1H).
Step a. This step was conducted in a similar manner to Example 39, step a, using Intermediate 5b and Intermediate B70.
m/z ES+ [M+H]+ 842.1; 1H NMR (400 MHz, CDCl3) δ ppm 8.31 (s, 1H), 7.67-7.65 (m, 4H), 7.46-7.39 (m, 7H), 7.35-7.28 (m, 3H), 7.05 (s, 1H), 6.80 (s, 1H), 4.70-4.63 (m, 1H), 4.35-4.22 (m, 1H), 4.20-4.09 (m, 2H), 4.05-4.01 (m, 1H), 3.94-3.85 (m, 1H), 3.84-3.69 (m, 3H), 3.68-3.56 (m, 1H), 3.34 (s, 3H), 3.29-3.21 (m, 1H), 3.08-2.95 (m, 2H), 2.74-2.63 (m, 1H), 2.49 (s, 3H), 2.25-2.13 (m, 1H), 1.08 (s, 9H).
Step b. This step was conducted in a similar manner to Example 38, step c.
m/z ES+ [M+H]+ 896.1.
Step c. To a solution of (3aR,11aS)-5-((7-acryloyl-6-(((tert-butyldiphenylsilyl)oxy)methyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin-2-yl)methyl)-6-chloro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (5 mg, 0.006 mmol) in THF (0.5 mL) was added TBAF (1 M in THF, 7 μL). The reaction mixture was stirred at 20° C. for 3 h. Upon completion, the reaction mixture was diluted with water (2 mL) and extracted with a mixture of DCM/2-propanol (3/1; 3×1 mL). The combined organic layers were dried over Na2SO4, filtered and evaporated. The residue was purified by prep-HPLC to give the title compound (1 mg, 27% yield) as a brown solid.
m/z ES+ [M+H]+ 658.2; 1H NMR (400 MHz, CDCl3) δ ppm 8.33 (s, 1H), 7.50-7.47 (m, 1H), 7.39-7.31 (m, 3H), 7.07 (s, 1H), 6.93-6.82 (m, 1H), 6.76-6.52 (m, 1H), 6.44 (d, J=16.4 Hz, 1H), 5.96-5.78 (m, 1H), 5.46-5.27 (m, 1H), 5.25-4.94 (m, 1H), 4.83-4.46 (m, 2H), 4.44-4.22 (m, 2H), 4.19-3.97 (m, 2H), 3.77-3.70 (m, 2H), 3.67-3.57 (m, 1H), 3.34 (d, J=3.2 Hz, 3H), 3.14-2.80 (m, 2H), 2.77-2.64 (m, 1H), 2.50 (s, 3H), 2.27-2.19 (m, 1H).
Step a. This step was conducted in a similar manner to Example 87, step a, using Intermediate 5b and 3-fluoro-4-nitrobenzaldehyde (CAS: 160538-51-2).
Step b. This step was conducted in a similar manner to Example 87, step c.
Step c. To a solution of (3aR,11aS)-5-(4-amino-3-fluorobenzyl)-6-chloro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (100 mg, 0.18 mmol) in pyridine (2.5 mL) was added T3P (340 mg, 0.53 mmol) and (E)-4-(dimethylamino)but-2-enoic acid hydrochloride (CAS: 848133-35-7; 35 mg, 0.21 mmol). The reaction mixture was stirred at 50° C. for 3 h. Upon completion, the mixture was diluted with water (10 mL) and extracted with EtOAc (3×5 mL). The combined organic layers were dried over Na2SO4, filtered and evaporated. The residue was purified by prep-HPLC to give the title compound (40 mg, 32% yield) as a pink solid.
m/z ES+ [M+H]+ 673.1; 1H NMR (400 MHz, CDCl3) δ ppm 8.37-8.28 (m, 2H), 7.48-7.45 (m, 2H), 7.30-7.27 (m, 2H), 7.25-7.24 (m, 1H), 7.13 (d, J=8.4 Hz, 1H), 7.05 (s, 1H), 7.01-6.97 (m, 1H), 6.20 (d, J=15.2 Hz, 1H), 4.68 (d, J=9.2 Hz, 1H), 4.23-4.16 (m, 1H), 4.12-4.06 (m, 1H), 3.51-3.44 (m, 1H), 3.36 (s, 3H), 3.19 (m, 2H), 3.04-2.91 (m, 2H), 2.67 (m, 1H), 2.48 (s, 3H), 2.34 (s, 6H), 2.22-2.15 (m, 1H).
The title compound was prepared in a similar manner to Example 245, using 2-fluoro-4-nitrobenzaldehyde (CAS: 157701-72-9) in step a.
m/z ES+ [M+H]+ 673.2; 1H NMR (400 MHz, CDCl3) δ ppm 8.43 (d, J=11.6 Hz, 2H), 8.30 (s, 1H), 7.57 (d, J=11.2 Hz, 1H), 7.47-7.44 (m, 1H), 7.29 (d, J=4.8 Hz, 2H), 7.26-7.23 (m, 1H), 7.04 (s, 1H), 6.95-6.88 (m, 1H), 6.32 (d, J=15.6 Hz, 1H), 4.64 (d, J=9.2 Hz, 1H), 4.33-4.11 (m, 2H), 3.56-3.52 (m, 1H), 3.42 (d, J=6.4 Hz, 2H), 3.24 (s, 3H), 3.00 (d, J=14.0 Hz, 1H), 2.95-2.84 (m, 1H), 2.71-2.65 (m, 1H), 2.54 (s, 6H), 2.47 (s, 3H), 2.23-2.12 (m, 1H).
The title compound was prepared in a similar manner to Example 245, using 3-nitrobenzaldehyde in step a.
m/z ES+ [M+H]+ 655.1; 1H NMR (400 MHz, CDCl3) δ ppm 8.33 (s, 1H), 7.75 (d, J=7.6 Hz, 1H), 7.51-7.38 (m, 3H), 7.35-7.28 (m, 3H), 7.07 (s, 2H), 6.989-6.94 (m, 1H), 6.13 (d, J=15.2 Hz, 1H), 4.66 (d, J=9.6 Hz, 1H), 4.33-4.10 (m, 2H), 3.51-3.46 (m, 1H), 3.30 (s, 3H), 3.12 (d, J=6.0 Hz, 2H), 3.07-2.98 (m, 1H), 2.96-2.84 (m, 1H), 2.68-2.61 (m, 1H), 2.50 (s, 3H), 2.28 (s, 6H), 2.20-2.13 (m, 1H).
Step a. This step was conducted in a similar manner to Example 87, step a, using Intermediate 5b and 3-bromo-4-nitrobenzaldehyde (CAS: 101682-68-2).
Step b. To a solution of (3aR,11aS)-5-(3-bromo-4-nitrobenzyl)-6-chloro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (200 mg, 0.31 mmol) and Zn(CN)2 (144 mg, 1.22 mmol) in DMA (1 mL) was added Pd(t-Bu3P)2 (CAS: 53199-31-8; 16 mg, 0.031 mmol). The mixture was purged with N2 3 times and stirred at 130° C. for 1 h under an N2 atmosphere. Upon completion, the reaction mixture was diluted with water (10 mL) and extracted with EtOAc (3×10 mL). The combined organic layers were washed with brine (2×20 mL), dried over Na2SO4, and evaporated. The residue was purified by reverse phase flash (water (0.1% NH4OH)/MeCN) to give 5-(((3aR,11aS)-6-chloro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)methyl)-2-nitrobenzonitrile (110 mg, 60% yield) as a white solid.
m/z ES+ [M+H]+ 599.2.
Step c. This step was conducted in a similar manner to Example 87, step c.
m/z ES+ [M+H]+ 569.2.
Step d. This step was conducted in a similar manner to Example 38, step c.
m/z ES+ [M+H]+ 623.1; 1H NMR (400 MHz, CDCl3) δ ppm 8.49 (d, J=8.8 Hz, 1H), 8.34 (s, 1H), 7.71-7.61 (m, 3H), 7.48 (t, J=4.8 Hz, 1H), 7.31 (d, J=4.4 Hz, 2H), 7.06 (s, 1H), 6.61-6.40 (m, 1H), 6.38-6.20 (m, 1H), 5.90 (d, J=10.4 Hz, 1H), 4.69 (d, J=9.6 Hz, 1H), 4.30-4.18 (m, 1H), 4.14-4.04 (m, 1H), 3.51-3.42 (m, 1H), 3.38 (s, 3H), 3.10-2.99 (m, 1H), 2.97-2.84 (m, 1H), 2.76-2.63 (m, 1H), 2.48 (s, 3H), 2.29-2.11 (m, 1H).
Steps a-c. These 3 steps were conducted as described in Example 87, steps a-c.
Step d. This step was conducted in a similar manner to Example 87, step d, using Intermediate E10.
m/z ES+ [M+H]+ 776.2; 1H NMR (400 MHz, CDCl3) δ ppm 8.33 (s, 1H), 7.61-7.51 (m, 2H), 7.35-7.30 (m, 3H), 7.27-7.23 (m, 4H), 7.05 (s, 1H), 7.03-6.93 (m, 1H), 4.77 (d, J=9.6 Hz, 1H), 4.16-4.12 (m, 1H), 4.05-3.93 (m, 1H), 3.70-3.60 (m, 3H), 3.55-3.15 (m, 7H), 3.05-2.90 (m, 1H), 2.76-2.63 (m, 5H), 2.50 (d, J=15.6 Hz, 7H), 2.21-2.10 (m, 1H), 1.49 (s, 9H).
Step e. To a solution of tert-butyl 4-((E)-4-((4-(((3aR,11aS)-6,10-dimethyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)methyl)phenyl)amino)-4-oxobut-2-en-1-yl)piperazine-1-carboxylate (90 mg, 0.12 mmol) in DCM (5 mL) was added TFA (1.54 g, 13.5 mmol). The reaction mixture was stirred at 25° C. for 1 h. Upon completion, the reaction mixture was basified to pH 8 with sat. aq. NaHCO3. The aqueous mixture was diluted with water (60 mL) and extracted with EtOAc (2×40 mL). The combined organic layers were washed with brine (3×30 mL), dried over Na2SO4 and evaporated to give (E)-N-(4-(((3aR,11aS)-6,10-dimethyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)methyl)phenyl)-4-(piperazin-1-yl)but-2-enamide (50 mg, 64% yield) as a white solid.
m/z ES+ [M+H]+ 676.1.
Step f. A mixture of (E)-N-(4-(((3aR,11aS)-6,10-dimethyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)methyl)phenyl)-4-(piperazin-1-yl)but-2-enamide (50 mg, 0.074 mmol), acetic acid (9 mg, 0.16 mmol), HATU (37 mg, 0.096 mmol) and DIPEA (24 mg, 0.19 mmol) in DCM (1 mL) was stirred at 25° C. for 12 h under a N2 atmosphere. Upon completion, the reaction mixture was evaporated. The residue was purified by reverse phase flash chromatography (water (0.1% TFA)/MeCN) followed by Prep-HPLC to give the title compound (27 mg, 47% yield) as a white solid.
m/z ES+ [M+H]+ 718.4; 1H NMR (400 MHz, CDCl3) δ ppm 8.32 (s, 1H), 7.56 (d, J=8.0 Hz, 2H), 7.47 (s, 1H), 7.32 (d, J=8.0 Hz, 2H), 7.25 (s, 3H), 7.05 (s, 1H), 7.01-6.92 (m, 1H), 6.17 (d, J=15.2 Hz, 1H), 4.76 (d, J=9.6 Hz, 1H), 4.13 (d, J=13.6 Hz, 1H), 3.96 (d, J=14.0 Hz, 1H), 3.68 (s, 2H), 3.58-3.48 (m, 2H), 3.46-3.36 (m, 4H), 3.24 (d, J=5.6 Hz, 2H), 3.05-2.91 (m, 1H), 2.76-2.59 (m, 2H), 2.53 (d, J=4.4 Hz, 2H), 2.50 (s, 5H), 2.46 (s, 3H), 2.20-2.08 (m, 4H).
Steps a. This step was conducted in a similar manner to Example 39, step a, using Intermediate 5b and Intermediate B75.
m/z ES+ [M+H]+ 686.3; 1H NMR (400 MHz, CDCl3) δ ppm 8.32 (s, 1H), 7.83 (s, 1H), 7.50-7.40 (m, 1H), 7.33-7.27 (m, 2H), 7.11-7.01 (m, 2H), 6.85 (d, J=8.4 Hz, 1H), 4.69 (d, J=9.2 Hz, 1H), 4.24 (t, J=4.4 Hz, 2H), 4.14-4.07 (m, 2H), 3.89-3.81 (m, 2H), 3.55-3.44 (m, 1H), 3.38 (s, 3H), 2.99-2.88 (m, 2H), 2.70-2.60 (m, 1H), 2.48 (s, 3H), 2.25-2.15 (m, 1H), 1.55 (s, 9H).
Step b. To a solution of tert-butyl 6-(((3aR,11aS)-6-chloro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)methyl)-2,3-dihydro-4H-benzo[b][1,4]oxazine-4-carboxylate (240 mg, 0.35 mmol) in DCM (12 mL) was added ZnBr2 (788 mg, 3.5 mmol). The mixture was stirred at 20° C. for 1 h. Upon completion, the reaction mixture was diluted with water (10 mL) and extracted with EtOAc (3×5 mL). The combined organic layers were dried over Na2SO4, and evaporated to give (3aR,11aS)-6-chloro-5-((3,4-dihydro-2H-benzo[b][1,4]oxazin-6-yl)methyl)-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (200 mg, crude) as a light yellow solid.
m/z ES+ [M+H]+ 586.2
Step c. This step was conducted in a similar manner to Example 38, step c.
m/z ES+ [M+H]+ 640.1; 1H NMR (400 MHz, CDCl3) δ ppm 8.33 (s, 1H), 7.51-7.43 (m, 1H), 7.34-7.28 (m, 2H), 7.23 (d, J=7.6 Hz, 1H), 7.14-7.03 (m, 2H), 6.89 (d, J=8.4 Hz, 1H), 6.72-6.61 (m, 1H), 6.57-6.49 (m, 1H), 5.82-5.79 (m, 1H), 4.68 (d, J=8.8 Hz, 1H), 4.33 (t, J=4.8 Hz, 2H), 4.18 (d, J=14.0 Hz, 1H), 4.09-3.95 (m, 3H), 3.56-3.42 (m, 1H), 3.35 (s, 3H), 3.02-2.90 (m, 2H), 2.71-2.67 (m, 1H), 2.48 (s, 3H), 2.24-2.20 (m, 1H).
The title compound was prepared in a similar manner to Example, using Intermediate 5b and Intermediate B75 in step a. The final step was conducted in a similar manner to the amide coupling described for Example 27.
m/z ES+ [M+H]+ 697.1; 1H NMR (400 MHz, CDCl3) δ ppm 8.32 (s, 1H), 7.47-7.45 (m, 1H), 7.33-7.27 (m, 3H), 7.11-7.10 (m, 1H), 7.07-7.01 (m, 2H), 6.89 (d, J=8.4 Hz, 1H), 6.60-6.53 (m, 1H), 4.69 (d, J=9.2 Hz, 1H), 4.36-4.26 (m, 2H), 4.18-4.12 (m, 1H), 4.11-4.04 (m, 1H), 4.03-3.98 (m, 1H), 3.98-3.90 (m, 1H), 3.55-3.44 (m, 1H), 3.36 (s, 3H), 3.15-3.05 (m, 2H), 3.00-2.88 (m, 2H), 2.75-2.60 (m, 1H), 2.48 (s, 3H), 2.24 (s, 6H), 2.23-2.14 (m, 1H).
Step a. This step was conducted in a similar manner to Example 83, step a, using Intermediate B82.
Steps b-c. These two steps were conducted in a similar manner to Example 83, steps c-d.
m/z ES+ [M+H]+ 621.3; 1H NMR (400 MHz, CDCl3) δ ppm 8.36 (s, 1H), 7.47-7.45 (m, 1H), 7.34-7.30 (m, 2H), 7.08 (s, 1H), 6.85-6.78 (m, 1H), 5.96 (d, J=15.6 Hz, 1H), 5.70-5.62 (m, 1H), 4.62 (d, J=9.2 Hz, 1H), 3.55-3.45 (m, 1H), 3.38 (s, 3H), 3.35-3.25 (m, 2H), 3.18-3.12 (m, 1H), 3.06 (d, J=5.6 Hz, 2H), 2.98-2.90 (m, 2H), 2.76-2.69 (m, 1H), 2.51 (s, 3H), 2.28 (s, 6H), 2.24-2.18 (m, 1H), 1.58-1.54 (m, 2H), 1.48-1.37 (m, 3H).
The title compound was prepared in a similar manner to Example 252, using Intermediate B86 in step a.
m/z ES+ [M+H]+ 653.2; 1H NMR (400 MHz, CDCl3) δ ppm 8.34 (s, 1H), 7.48-7.45 (m, 1H), 7.35-7.27 (m, 2H), 7.07 (s, 1H), 6.87-6.76 (m, 1H), 6.16-5.98 (m, 2H), 4.58 (d, J=9.2 Hz, 1H), 4.53-4.32 (m, 1H), 3.62-3.44 (m, 2H), 3.43-3.29 (m, 4H), 3.29-2.89 (m, 6H), 2.77-2.66 (m, 1H), 2.50 (s, 3H), 2.26-2.15 (m, 7H), 1.83-1.74 (m, 2H), 1.67-1.42 (m, 2H).
Step a. This step was conducted in a similar manner to Example 79, step a, using Intermediate 5b and Intermediate B30.
m/z ES+ [M+H]+ 650.1; 1H NMR (400 MHz, CDCl3) δ ppm 8.33 (s, 1H), 7.46-7.43 (m, 1H), 7.32-7.27 (m, 2H), 7.05 (s, 1H), 4.63 (d, J=9.2 Hz, 1H), 4.00-3.92 (m, 2H), 3.58-3.45 (m, 3H), 3.36 (s, 3H), 3.05-2.87 (m, 4H), 2.85-2.70 (m, 1H), 2.48 (s, 3H), 2.46-2.39 (m, 1H), 2.25-2.20 (m, 1H), 1.56-1.49 (m, 2H), 1.43 (s, 9H), 1.39-1.30 (m, 2H), 1.29-1.25 (m, 2H).
Step b. This step was conducted in a similar manner to Example 77, step b.
m/z ES+ [M+H]+ 630.1.
Steps c-f. These 4 steps were conducted in a similar manner to Example 79, steps b-e, using Intermediate E3 in step d.
m/z ES+ [M+H]+ 675.3; 1H NMR (400 MHz, CDCl3) δ ppm 8.33 (s, 1H) 7.16-7.26 (m, 3H) 7.04 (s, 1H) 6.71-6.63 (m, 1H) 4.70 (d, J=9.2 Hz, 1H) 4.50-4.35 (m, 1H) 4.20-4.06 (m, 1H) 3.87-4.00 (m, 1H) 3.71-3.62 (m, 1H) 3.61-3.51 (m, 1H) 3.35 (s, 3H) 3.21 (d, J=6.4 Hz, 2H) 2.84-3.01 (m, 3H) 2.67-2.83 (m, 2H) 2.51-2.61 (m, 1H) 2.49 (s, 3H) 2.37 (s, 3H) 2.28 (s, 6H) 2.23-2.14 (m, 1H) 1.51-1.63 (m, 2H) 1.22-1.39 (m, 4H).
The title compound was prepared in a similar manner to Example 254, using Intermediate 5b and Intermediate B18 in step a.
m/z ES+ [M+H]+ 695.7; 1H NMR (400 MHz, CDCl3) δ ppm 8.32 (s, 1H), 7.29 (s, 3H), 7.25 (s, 3H), 7.06 (s, 1H), 6.72 (t, J=6.4 Hz, 1H), 4.97 (s, 2H), 4.88 (s, 2H), 4.77-4.74 (m, 1H), 4.20-4.15 (m, 1H), 4.08-3.99 (m, 1H), 3.94 (d, J=6.8 Hz, 2H), 3.46-3.39 (m, 4H), 3.04-2.95 (m, 1H), 2.90 (s, 6H), 2.79-2.62 (m, 2H), 2.51 (s, 3H), 2.47 (d, J=3.2 Hz, 3H), 2.20-2.09 (m, 1H).
Step a. This step was conducted in a similar manner to Example 1, step a, using Intermediate 7b and tert-butyl piperazine-1-carboxylate.
m/z ES+ [M+H]+ 651.3.
Step b. This step was conducted in a similar manner to Example 77, step b.
m/z ES+ [M+H]+ 631.3.
Steps c-d. These 2 steps were conducted in a similar manner to Example 1, steps b-c.
m/z ES+ [M+H]+ 585.3; 1H NMR (400 MHz, CDCl3) δ ppm 8.33 (s, 1H), 7.26-7.18 (m, 3H), 7.05 (s, 1H), 6.61-6.46 (m, 1H), 6.29 (d, J=16.0 Hz, 1H), 5.70 (d, J=10.4 Hz, 1H), 4.70 (d, J=9.6 Hz, 1H), 3.79-3.44 (m, 5H), 3.38 (s, 3H), 3.30-3.03 (m, 2H), 3.03-2.92 (m, 1H), 2.86-2.77 (m, 1H), 2.76-2.68 (m, 1H), 2.49 (s, 3H), 2.49-2.40 (m, 4H), 2.40 (s, 3H), 2.39-2.26 (m, 2H), 2.24-2.15 (m, 1H).
Step a. To a mixture of Intermediate 5b (2 g, 4.56 mmol), 1,4-dioxane-2,5-diol (CAS: 23147-58-2; 2.46 g, 2.05 mmol), 4 Å molecular sieves (1 g) and acetic acid (1.37 g, 22.8 mmol) in MeOH (20 mL) was added NaBH3CN (1.00 g, 16.0 mmol) at 0° C. The reaction mixture was stirred at 20° C. for 16 h under a N2 atmosphere. Upon completion, the reaction mixture was filtered and evaporated. The residue was purified by column chromatography (PE/EtOAc=1/2) to give (3aR,11aS)-6-chloro-5-(2-hydroxyethyl)-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (1.7 g, 77% yield) as light yellow solid.
m/z ES+ [M+H]+ 483.0; 1H NMR (400 MHz, CDCl3) δ ppm 8.34 (s, 1H), 7.49-7.47 (m, 1H), 7.37-7.30 (m, 2H), 7.06 (s, 1H), 4.67 (d, J=9.2 Hz, 1H), 3.67-3.55 (m, 2H), 3.51-3.42 (m, 1H), 3.39 (s, 3H), 3.31-3.19 (m, 1H), 3.10-3.06 (m, 1H), 3.05-2.91 (m, 2H), 2.80-2.71 (m, 1H), 2.49 (s, 4H), 2.50-2.26 (m, 1H).
Step b. A mixture of (3aR,11aS)-6-chloro-5-(2-hydroxyethyl)-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (400 mg, 0.83 mmol), methanesulfonic anhydride (289 mg, 1.66 mmol) and TEA (838 mg, 8.28 mmol) in DCM (5 mL) was stirred at 25° C. for 3 h under a N2 atmosphere. Upon completion, the reaction mixture was washed with water (3×15 mL) and brine (15 mL), dried over Na2SO4 and evaporated to give 2-((3aR,11aS)-6-chloro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)ethyl methanesulfonate (0.45 g, crude) as a yellow oil, which was used without further purification.
1H NMR (400 MHz, CDCl3) δ ppm 8.26 (s, 1H), 7.41-7.39 (m, 1H), 7.34-7.24 (m, 2H), 6.99 (s, 1H), 4.54 (d, J=9.6 Hz, 1H), 4.15-4.05 (m, 1H), 4.03-3.93 (m, 1H), 3.55-3.48 (m, 1H), 3.44-3.34 (m, 3H), 3.32 (s, 3H), 2.98 (s, 3H), 2.91-2.81 (m, 1H), 2.73 (s, 3H), 2.67-2.65 (m, 1H), 2.18-2.11 (m, 1H).
Step c. A mixture of 2-((3aR,11aS)-6-chloro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)ethyl methanesulfonate (300 mg, 0.54 mmol), tert-butyl (3,3-difluoropiperidin-4-yl)carbamate (CAS: 1255666-48-8; 316 mg, 1.34 mmol) and TEA (162 mg, 1.60 mmol) in EtOH (1 mL) was stirred at 80° C. for 1 h under a N2 atmosphere. Upon completion, the reaction mixture was evaporated. The residue was redissolved with EtOAc (15 mL) and washed with water (20 mL) and brine (3×15 mL), dried over Na2SO4 and evaporated. The residue was purified by column chromatography (PE/EtOAc=3/1 to 1/1) to give tert-butyl (1-(2-((3aR,11aS)-6-chloro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)ethyl)-3,3-difluoropiperidin-4-yl)carbamate (200 mg, 53% yield) as a white solid.
m/z ES+ [M+H]+ 701.3; 1H NMR (400 MHz, CDCl3) δ ppm 8.26 (s, 1H), 7.38-7.36 (m, 1H), 7.27-7.21 (m, 2H), 6.97 (s, 1H), 4.76-4.65 (m, 2H), 4.52 (t, J=9.2 Hz, 1H), 3.56-3.44 (m, 1H), 3.29 (d, J=5.2 Hz, 3H), 3.23-3.13 (m, 2H), 3.06 (t, J=6.8 Hz, 1H), 3.02-2.96 (m, 2H), 2.95-2.84 (m, 2H), 2.82-2.72 (m, 2H), 2.64-2.54 (m, 2H), 2.41 (d, J=1.2 Hz, 3H), 2.39-2.34 (m, 1H), 2.17-2.07 (m, 2H), 1.39 (s, 9H).
Steps d-e. These two steps were conducted in a similar manner to Example 37a and Example 37b, steps c-d.
Example 257a:
m/z ES+ [M+H]+ 655.2; 1H NMR (400 MHz, CDCl3) δ ppm 8.36 (s, 1H), 7.48-7.46 (m, 1H), 7.34-7.31 (m, 2H), 7.07 (s, 1H), 6.38-6.33 (m, 1H), 6.19-6.10 (m, 1H), 5.77-5.69 (m, 2H), 4.63 (d, J=9.2 Hz, 1H), 4.41-4.23 (m, 1H), 3.64 (d, J=9.6 Hz, 1H), 3.39 (s, 3H), 3.21-3.14 (m, 2H), 3.08-2.88 (m, 4H), 2.77-2.73 (m, 1H), 2.50 (s, 3H), 2.47-2.33 (m, 2H), 2.29-2.15 (m, 3H), 2.08-1.97 (m, 1H), 1.70-1.61 (m, 1H).
Example 257b:
m/z ES+ [M+H]+ 655.2; 1H NMR (400 MHz, CDCl3) δ ppm 8.35 (s, 1H), 7.49-7.46 (m, 1H), 7.37-7.31 (m, 2H), 7.07 (s, 1H), 6.38-6.34 (m, 1H), 6.20-6.12 (m, 1H), 5.79-5.72 (m, 2H), 4.60 (d, J=9.6 Hz, 1H), 4.40-4.25 (m, 1H), 3.60-3.56 (m, 1H), 3.40 (s, 3H), 3.36-3.18 (m, 2H), 3.16-2.96 (m, 3H), 2.94-2.88 (m, 1H), 2.75-2.71 (m, 1H), 2.51 (s, 3H), 2.48-2.32 (m, 2H), 2.30-2.17 (m, 3H), 2.05-1.97 (m, 1H), 1.74-1.68 (m, 1H).
The title compound was prepared in a similar manner to Example 257a and Example 257b, using tert-butyl 2-(cyanomethyl)piperazine-1-carboxylate (CAS: 1808997-73-0) in step c.
Example 258a:
m/z ES+ [M+H]+ 644.3; 1H NMR (400 MHz, CDCl3) δ ppm 8.35 (s, 1H), 7.48-7.43 (m, 1H), 7.35-7.29 (m, 2H), 7.06 (s, 1H), 6.61-6.43 (m, 1H), 6.39-6.27 (m, 1H), 5.82-5.68 (m, 1H), 5.06-4.87 (m, 1H), 4.64 (d, J=9.2 Hz, 1H), 3.82-3.71 (m, 1H), 3.65-3.53 (m, 1H), 3.40 (s, 3H), 3.27-3.05 (m, 3H), 3.03-2.89 (m, 4H), 2.79-2.72 (m, 2H), 2.69-2.58 (m, 1H), 2.48 (s, 3H), 2.46-2.32 (m, 2H), 2.30-2.15 (m, 2H), 2.13-1.95 (m, 1H).
Example 258b:
m/z ES+ [M+H]+ 644.3; 1H NMR (400 MHz, CDCl3) δ ppm 8.34 (s, 1H), 7.53-7.47 (m, 1H), 7.38-7.30 (m, 2H), 7.06 (s, 1H), 6.63-6.45 (m, 1H), 6.39-6.29 (m, 1H), 5.76 (d, J=10.4 Hz, 1H), 5.03-4.86 (m, 1H), 4.63 (d, J=8.8 Hz, 1H), 3.88-3.71 (m, 1H), 3.61 (d, J=9.6 Hz, 1H), 3.41 (s, 3H), 3.25-3.15 (m, 2H), 3.07-2.88 (m, 5H), 2.87-2.81 (m, 1H), 2.66-2.53 (m, 1H), 2.49 (s, 3H), 2.46-2.34 (m, 2H), 2.32-2.12 (m, 3H), 1.96-1.81 (m, 1H).
The title compound was prepared in a similar manner to Example 257a and Example 257b, using Intermediate C5 in step c.
m/z ES+ [M+H]+ 628.2; 1H NMR (400 MHz, CDCl3) δ ppm 8.34 (s, 1H), 7.48-7.44 (m, 1H), 7.36 (s, 2H), 7.34 (d, J=1.6 Hz, 1H), 7.06 (s, 1H), 6.55-6.44 (m, 2H), 5.84-5.75 (m, 1H), 4.67-4.60 (m, 4H), 4.59-4.54 (m, 1H), 3.97-3.84 (m, 2H), 3.56-3.46 (m, 1H), 3.38 (s, 3H), 3.35-3.30 (m, 1H), 3.09-2.98 (m, 1H), 2.95-2.83 (m, 1H), 2.80-2.72 (m, 1H), 2.48 (s, 3H), 2.27-2.21 (m, 1H), 2.08-1.96 (m, 1H).
Step a. This step was conducted in a similar manner to Example 257a and Example 257b, step a, using Intermediate 6c.
m/z ES+ [M+H]+ 463.1; 1H NMR (400 MHz, CDCl3) δ ppm 8.33 (s, 1H), 7.27-7.22 (m, 3H), 7.05 (s, 1H), 4.71 (d, J=9.6 Hz, 1H), 3.69-3.48 (m, 3H), 3.37 (s, 3H), 3.25-3.15 (m, 1H), 3.15-3.06 (m, 1H), 3.05-2.92 (m, 1H), 2.91-2.81 (m, 1H), 2.74 (dd, J=16.8, 8.4 Hz, 1H), 2.50 (s, 3H), 2.42 (s, 3H), 2.21 (dd, J=16.8, 11.2 Hz, 1H), 2.06 (br s, 1H).
Step b. To a solution of (3aR,11aS)-5-(2-hydroxyethyl)-6,10-dimethyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (1.1 g, 2.38 mmol) in DCM (12 mL) was added imidazole (259 mg, 3.81 mmol), PPh3 (936 mg, 3.57 mmol) and I2 (905 mg, 3.57 mmol). The mixture was stirred at 25° C. for 12 h under a N2 atmosphere. Upon completion, the mixture was diluted with water (40 mL) and extracted with DCM (2×40 mL). The combined organic layers were washed with brine (40 mL), dried over Na2SO4 and evaporated. Purification by column chromatography (PE/EtOAc=3/1) afforded (3aR,11aS)-5-(2-iodoethyl)-6,10-dimethyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (0.74 g, 54% yield) as a white solid.
m/z ES+ [M+H]+ 573.2; 1H NMR (400 MHz, CDCl3) δ ppm 8.34 (s, 1H), 7.47 (d, J=2.8 Hz, 1H), 7.26-7.23 (m, 2H), 7.05 (s, 1H), 4.71 (d, J=9.2 Hz, 1H), 3.6-3.7 (m, 1H), 3.39 (s, 3H), 3.38-3.33 (m, 1H), 3.29-3.20 (m, 1H), 3.15-3.00 (m, 2H), 3.00-2.89 (m, 1H), 2.87-2.70 (m, 2H), 2.49 (s, 3H), 2.46 (s, 3H), 2.1-2.3 (m, 1H).
Step c. To a solution of (3aR,11aS)-5-(2-iodoethyl)-6,10-dimethyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (180 mg, 0.31 mmol) in MeCN (2 mL) was added DIPEA (81 mg, 0.63 mmol) and tert-butyl (S)-3-methylpiperazine-1-carboxylate (CAS: 147081-29-6; 95 mg, 0.47 mmol). The mixture was stirred at 60° C. for 12 h. Upon completion, water (30 ml) was added and the aqueous mixture was extracted with EtOAc (3×30 ml). The combined organic layers were washed with brine (75 mL), dried over Na2SO4 and evaporated. Purification by column chromatography (EtOAc) afforded tert-butyl (S)-4-(2-((3aR,11aS)-6,10-dimethyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)ethyl)-3-methylpiperazine-1-carboxylate (0.2 g, 91% yield) as a white solid.
m/z ES+ [M+H]+ 645.4; 1H NMR (400 MHz, CDCl3) δ ppm 8.33 (s, 1H), 7.26-7.19 (m, 3H), 7.06-7.03 (m, 1H), 4.70 (d, J=9.6 Hz, 1H), 3.77-3.51 (m, 3H), 3.38 (s, 3H), 3.27-2.94 (m, 4H), 2.91-2.66 (m, 5H), 2.49 (s, 3H), 2.40 (s, 3H), 2.37-2.27 (m, 1H), 2.24-2.08 (m, 3H), 1.45 (s, 9H), 0.90 (d, J=6.4 Hz, 3H).
Steps d-e. These 2 steps were conducted in a similar manner to Example 1, steps b-c.
m/z ES+ [M+H]+ 599.3; 1H NMR (400 MHz, CDCl3) δ ppm 8.34 (s, 1H), 7.27-7.20 (m, 3H), 7.06 (s, 1H), 6.61-6.50 (m, 1H), 6.24-6.35 (m, 1H), 5.70 (d, J=10.8 Hz, 1H), 4.71 (d, J=9.6 Hz, 1H), 4.15-3.86 (m, 1H), 3.77-3.54 (m, 2H), 3.39 (s, 4H), 3.18-2.63 (m, 8H), 2.51 (s, 3H), 2.41 (s, 4H), 2.27-2.14 (m, 3H), 0.94 (s, 3H).
The title compound was prepared in a similar manner to Example 260, using tert-butyl 3,6-diazabicyclo[3.1.1]heptane-3-carboxylate (CAS: 1251017-66-9) in step c.
m/z ES+ [M+H]+ 597.2; 1H NMR (400 MHz, CDCl3) δ ppm 8.32 (s, 1H), 7.25-7.17 (m, 3H), 7.04 (s, 1H), 6.62-6.51 (m, 1H), 6.49-6.39 (m, 1H), 5.7-5.85 (m, 1H), 4.65-4.75 (m, 1H), 3.78 (d, J=11.2 Hz, 1H), 3.69-3.50 (m, 6H), 3.35 (d, J=8.8 Hz, 3H), 3.07-2.85 (m, 3H), 2.84-2.67 (m, 2H), 2.62-2.51 (m, 1H), 2.48 (s, 3H), 2.47-2.43 (m, 1H), 2.41 (s, 1H), 2.37 (s, 3H), 2.1-2.25 (m, 1H), 1.48 (d, J=8.8 Hz, 1H).
The title compound was prepared in a similar manner to Example 260, using tert-butyl 3,8-diazabicyclo[3.2.1]octane-3-carboxylate (CAS: 201162-53-0) in step c.
m/z ES+ [M+H]+ 611.2; 1H NMR (400 MHz, CDCl3) δ ppm 8.33 (s, 1H), 7.26-7.19 (m, 3H), 7.05 (s, 1H), 6.55-6.44 (m, 1H), 6.2-6.35 (m, 1H), 5.67 (d, J=10.4 Hz, 1H), 4.74-4.67 (m, 1H), 4.23 (d, J=13.2 Hz, 1H), 3.72-3.49 (m, 2H), 3.38 (d, J=3.6 Hz, 3H), 3.35-3.21 (m, 1H), 3.20-2.92 (m, 5H), 2.91-2.66 (m, 3H), 2.49 (s, 3H), 2.41 (s, 3H), 2.35 (d, J=6.0 Hz, 2H), 2.15-2.25 (m, 1H), 1.84 (s, 2H), 1.62-1.47 (m, 2H).
The title compound was prepared in a similar manner to Example 260, using tert-butyl (1R,4R)-2,5-diazabicyclo[2.2.1]heptane-2-carboxylate (CAS: 134003-84-2) in step c.
m/z ES+ [M+H]+ 597.2; 1H NMR (400 MHz, CDCl3) δ ppm 8.33 (s, 1H), 7.26-7.18 (m, 3H), 7.04 (s, 1H), 6.48-6.26 (m, 2H), 5.69 (dd, J=9.6, 2.4 Hz, 1H), 4.81-4.75 (m, 1H), 4.69 (dd, J=9.6, 4.8 Hz, 1H), 3.70-3.46 (m, 3H), 3.36 (d, J=9.2 Hz, 4H), 3.08-2.76 (m, 5H), 2.72 (dd, J=16.8, 8.4 Hz, 1H), 2.66-2.53 (m, 2H), 2.49 (s, 3H), 2.49-2.39 (m, 1H), 2.37 (s, 3H), 2.24-2.13 (m, 1H), 1.68-1.61 (m, 2H).
Steps a-b. These 2 steps were conducted in a similar manner to Example 257a and Example 257b, steps a-b, using Intermediate 6c in step a.
m/z ES+ [M+H]+ 541.1; 1H NMR (400 MHz, CDCl3) δ ppm 8.33 (s, 1H), 7.27-7.22 (m, 3H), 7.05 (s, 1H), 4.71 (d, J=9.6 Hz, 1H), 4.15-4.10 (m, 1H), 3.68-3.50 (m, 3H), 3.39-3.30 (m, 4H), 3.21-3.02 (m, 1H), 3.00 (s, 3H), 2.96-2.73 (m, 2H), 2.49 (s, 3H), 2.42 (s, 3H), 2.21 (dd, J=16.8, 11.4 Hz, 1H).
Step c. A solution of 2-((3aR,11aS)-6,10-dimethyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)ethyl methanesulfonate (800 mg, 1.48 mmol) and piperazine (1.27 g, 14.8 mmol) in MeCN (10 mL) was stirred at 50° C. for 12 h. Upon completion, the mixture was evaporated. The residue was purified by prep-HPLC to give (3aR,11aS)-6,10-dimethyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-5-(2-(piperazin-1-yl)ethyl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (700 mg, 89% yield) as a yellow solid.
m/z ES+ [M+H]+ 531.3; 1H NMR (400 MHz, CDCl3) δ ppm 8.32 (s, 1H), 7.27-7.18 (m, 3H), 7.04 (s, 1H), 4.70 (d, J=9.6 Hz, 1H), 3.61 (dd, J=14.2, 4.4 Hz, 1H), 3.37 (s, 3H), 3.20-2.95 (m, 3H), 2.88-2.85 (m, 4H), 2.82-2.68 (m, 2H), 2.49 (s, 3H), 2.39-2.32 (m, 9H), 2.18 (dd, J=16.8, 11.6 Hz, 2H).
Steps d. This step was conducted in a similar manner to Example 37a and Example 37b, step d, using propiolic acid.
m/z ES+ [M+H]+ 583.3; 1H NMR (400 MHz, CDCl3) ppm 8.33 (s, 1H), 7.27-7.19 (m, 3H), 7.05 (s, 1H), 4.70 (d, J=9.6 Hz, 1H), 3.78-3.71 (m, 2H), 3.69-3.53 (m, 3H), 3.38 (s, 3H), 3.17 2.92 (m, 4H), 2.86-2.68 (m, 2H), 2.49 (s, 3H), 2.47-2.41 (m, 3H), 2.40 (s, 6H), 2.31-2.02 (m, 1H).
Step a. This step was conducted in a similar manner to Example 38, step a, using Intermediate 5b and Intermediate B87.
m/z ES+ [M+H]+ 587.0; 1H NMR (400 MHz, CDCl3) δ ppm 8.31 (s, 1H), 7.43-7.39 (m, 1H), 7.34-7.28 (m, 4H), 7.26 (d, J=6.4 Hz, 3H), 7.03 (s, 1H), 4.61 (d, J=9.2 Hz, 1H), 4.44 (s, 2H), 3.51 (t, J=6.0 Hz, 3H), 3.33 (s, 3H), 3.18-3.04 (m, 2H), 3.00-2.87 (m, 2H), 2.73-2.65 (m, 1H), 2.46 (s, 3H), 2.24-2.14 (m, 1H), 1.67-1.58 (m, 2H).
Step b. This step was conducted in a similar manner to Example 77, step b.
m/z ES+ [M+H]+ 567.3; 1H NMR (400 MHz, CDCl3) δ ppm 8.33 (s, 1H), 7.36-7.27 (m, 5H), 7.24-7.17 (m, 3H), 7.04 (s, 1H), 4.70 (d, J=9.6 Hz, 1H), 4.46 (s, 2H), 3.60-3.53 (m, 1H), 3.51-3.44 (m, 2H), 3.35 (s, 3H), 3.12-2.91 (m, 3H), 2.81-2.67 (m, 2H), 2.49 (s, 3H), 2.33 (s, 3H), 2.22-2.13 (m, 1H), 1.75-1.59 (m, 2H).
Step c. To a solution of (3aR,11aS)-5-(3-(benzyloxy)propyl)-6,10-dimethyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (350 mg, 0.62 mmol) in THF (10 mL) was added 10% Pd/C (100 mg). The reaction mixture was stirred at 25° C. for 32 h under a H2 atmosphere (15 psi). Upon completion, the reaction mixture was filtered through a pad of Celite and the solids were washed with EtOAc (2×10 mL). The filtrate were evaporated to give (3aR,11aS)-5-(3-hydroxypropyl)-6,10-dimethyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (290 mg, 98% yield) as a yellow solid.
m/z ES+ [M+H]+ 477.1; 1H NMR (400 MHz, CDCl3) δ ppm 8.33 (s, 1H), 7.26-7.18 (m, 3H), 7.04 (s, 1H), 4.71 (d, J=9.6 Hz, 1H), 3.69 (t, J=6.2 Hz, 2H), 3.65-3.58 (m, 1H), 3.38 (s, 3H), 3.17-2.94 (m, 3H), 2.85-2.68 (m, 2H), 2.49 (s, 3H), 2.38 (s, 3H), 2.24-2.15 (m, 1H), 1.98-1.95 (m, 1H), 1.69-1.61 (m, 2H).
Step d. This step was conducted in a similar manner to Example 257a and Example 257b, step b.
m/z ES+ [M+H]+ 555.2; 1H NMR (400 MHz, CDCl3) δ ppm 8.33 (s, 1H), 7.27-7.18 (m, 3H), 7.05 (s, 1H), 4.69 (d, J=9.6 Hz, 1H), 4.31-4.20 (m, 2H), 3.64-3.54 (m, 1H), 3.38 (s, 3H), 3.15-3.08 (m, 2H), 2.99 (s, 3H), 2.97-2.89 (m, 1H), 2.88-2.79 (m, 1H), 2.78-2.70 (m, 1H), 2.49 (s, 3H), 2.39 (s, 3H), 2.25-2.16 (m, 1H), 1.84-1.80 (m, 2H).
Step e. To a mixture of 3-((3aR,11aS)-6,10-dimethyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)propyl methanesulfonate (280 mg, 0.50 mmol) and 2-(trifluoromethyl)piperazine (CAS: 131922-05-9; 233 mg, 1.51 mmol) in EtOH (4 mL) was added TEA (153 mg, 1.51 mmol). The reaction mixture was stirred at 80° C. for 4 h. Additional 2-(trifluoromethyl)piperazine (78 mg, 0.50 mmol) was added. The reaction mixture was stirred at 80° C. for a further 2 h. Upon completion, the reaction mixture was diluted with water (10 mL) and extracted with EtOAc (3×10 mL). The combined organic layers were washed with brine (2×20 mL), dried over Na2SO4 and evaporated. Purification by reverse phase flash chromatography (water (0.1% NH4OH)/MeCN) afforded (3aR,11aS)-6,10-dimethyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-5-(3-(3-(trifluoromethyl)piperazin-1-yl)propyl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (200 mg, 65% yield) as a white solid.
m/z ES+ [M+H]+ 613.3; 1H NMR (400 MHz, CDCl3) δ ppm 8.33 (s, 1H), 7.26-7.16 (m, 3H), 7.04 (s, 1H), 4.74-4.67 (m, 1H), 3.65-3.53 (m, 1H), 3.39-3.35 (m, 3H), 3.35-3.26 (m, 1H), 3.08-2.79 (m, 7H), 2.78-2.68 (m, 2H), 2.49 (s, 3H), 2.39-2.38 (m, 5H), 2.24-2.15 (m, 1H), 2.09-1.97 (m, 2H), 1.61-1.51 (m, 3H).
Step f. This step was conducted in a similar manner to Example 38, step c. An additional chiral SFC step afforded the title compounds.
Example 265a:
m/z ES+ [M+H]+ 667.0; 1H NMR (400 MHz, CDCl3) δ ppm 8.33 (s, 1H), 7.26-7.19 (m, 3H), 7.04 (s, 1H), 6.61-6.46 (m, 1H), 6.40-6.30 (m, 1H), 5.82-5.74 (m, 1H), 5.37-5.09 (m, 1H), 4.69 (d, J=9.6 Hz, 1H), 4.63-4.20 (m, 1H), 3.86-3.48 (m, 2H), 3.36 (s, 3H), 3.14 (d, J=12.4 Hz, 1H), 3.11-3.03 (m, 1H), 3.02-2.91 (m, 2H), 2.90-2.77 (m, 2H), 2.76-2.68 (m, 1H), 2.49 (s, 3H), 2.43-2.30 (m, 5H), 2.25-2.09 (m, 2H), 2.06-1.93 (m, 1H), 1.59-1.49 (m, 2H).
Example 265b:
m/z ES+ [M+H]+ 667.0; 1H NMR (400 MHz, CDCl3) δ ppm 8.33 (s, 1H), 7.26-7.19 (m, 3H), 7.04 (s, 1H), 6.63-6.45 (m, 1H), 6.40-6.30 (m, 1H), 5.78 (d, J=10.8 Hz, 1H), 5.40-5.15 (m, 1H), 4.69 (d, J=9.6 Hz, 1H), 4.61-4.18 (m, 1H), 3.85-3.51 (m, 2H), 3.36 (s, 3H), 3.20 (d, J=12.8 Hz, 1H), 3.15-3.05 (m, 1H), 3.03-2.90 (m, 2H), 2.87-2.77 (m, 2H), 2.76-2.68 (m, 1H), 2.49 (s, 3H), 2.38 (s, 3H), 2.38-2.27 (m, 2H), 2.25-2.08 (m, 2H), 2.04-1.92 (m, 1H), 1.57-1.47 (m, 2H).
Step a. This step was conducted in a similar manner to Intermediate 7a, step a, using Intermediate 5b.
m/z ES+ [M+H]+ 479.0; 1H NMR (400 MHz, CDCl3) δ ppm 8.30 (s, 1H), 7.43-7.37 (m, 1H), 7.27-7.21 (m, 2H), 7.02 (s, 1H), 5.79-5.64 (m, 1H), 5.22-5.13 (m, 1H), 5.09-5.00 (m, 1H), 4.62 (d, J=9.2 Hz, 1H), 3.67-3.48 (m, 3H), 3.34 (s, 3H), 2.99-2.86 (m, 2H), 2.74-2.63 (m, 1H), 2.45 (s, 3H), 2.22-2.11 (m, 1H).
Step b. To a solution of (3aR,11aS)-5-allyl-6-chloro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (2 g, 4.18 mmol) in THF (20 mL) was added a solution of borane dimethyl sulfide complex (10 M in THF, 1.25 mL) in DCM (11 mL). The reaction mixture was stirred at 0° C. for 2 h. Upon completion, the reaction mixture was diluted with MeOH (10 mL) and evaporated. The residue was dissolved in THF (20 mL) and NaOH (501 mg, 12.5 mmol) and H2O2 (4.26 g, 37.6 mmol) were added. The resulting mixture was stirred at 0° C. for 1 h. Upon completion, the reaction mixture was quenched with sat. aq. Na2S2O3 (100 mL). The aqueous mixture was extracted with EtOAc (3×50 mL). The combined organic layers were washed with water (2×100 mL), sat. aq. Na2S2O3 (100 mL), brine (100 mL), dried over Na2SO4 and evaporated. The residue was purified by reverse phase flash chromatography (water (0.1% NH4OH)/MeCN) to give (3aR,11aS)-6-chloro-5-(3-hydroxypropyl)-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (1.3 g, 63% yield) as a yellow oil.
m/z ES+ [M+H]+ 497.2; 1H NMR (400 MHz, CDCl3) δ ppm 8.25 (s, 1H), 7.42-7.35 (m, 1H), 7.28-7.19 (m, 2H), 7.00-6.95 (m, 1H), 4.62-4.52 (m, 1H), 3.69-3.59 (m, 2H), 3.58-3.47 (m, 1H), 3.35-3.28 (m, 3H), 3.19-2.98 (m, 2H), 2.93-2.88 (m, 1H), 2.71-2.61 (m, 1H), 2.43-2.38 (m, 3H), 2.23-2.08 (m, 1H), 1.69-1.41 (m, 2H), 1.08-1.00 (m, 1H).
Step c. This step was conducted in a similar manner to Example 257a and Example 257b, step b.
m/z ES+ [M+H]+ 575.2.
Step d. To a solution of 3-((3aR,11aS)-6-chloro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)propyl methanesulfonate (190 mg, 0.33 mmol) in EtOH (2 mL) was added piperazine (285 mg, 3.30 mmol). The reaction mixture was stirred for 1 h at 80° C. Upon completion, the mixture was quenched with water (30 mL) and extracted with EtOAc (3×30 mL). The combined organic layers were washed with brine (30 mL), dried over Na2SO4 and evaporated to give (3aR,11aS)-6-chloro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-5-(3-(piperazin-1-yl)propyl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (140 mg, crude) as a yellow solid.
m/z ES+ [M+H]+ 565.3; 1H NMR (400 MHz, CDCl3) δ ppm 8.33 (s, 1H), 7.46-7.42 (m, 1H), 7.33-7.27 (m, 2H), 7.05 (s, 1H), 4.63 (d, J=9.2 Hz, 1H), 3.62-3.50 (m, 1H), 3.38 (s, 3H), 3.06 (t, J=6.8 Hz, 2H), 3.01-2.92 (m, 2H), 2.86 (t, J=4.8 Hz, 4H), 2.78-2.67 (m, 1H), 2.48 (s, 3H), 2.43-2.31 (m, 7H), 2.26-2.18 (m, 1H), 1.54 (t, J=7.2 Hz, 2H).
Step e. This step was conducted in a similar manner to Example 38, step c.
m/z ES+ [M+H]+ 619.1; 1H NMR (400 MHz, CDCl3) δ ppm 8.32 (s, 1H), 7.49-7.41 (m, 1H), 7.35-7.29 (m, 2H), 7.05 (s, 1H), 6.58-6.47 (m, 1H), 6.35-6.24 (m, 1H), 5.81-5.67 (m, 1H), 4.61 (d, J=9.2 Hz, 1H), 3.82-3.56 (m, 5H), 3.39 (s, 3H), 3.30-3.15 (m, 1H), 3.15-2.85 (m, 5H), 2.78-2.72 (m, 2H), 2.71-2.47 (m, 6H), 2.29-2.17 (m, 1H), 1.79-1.59 (m, 2H).
Steps a-c. These 3 steps were conducted as described in Example 266, steps a-c.
Steps d-f. These 3 steps were conducted in a similar manner to Example 257a and Example 257b, steps c-e, using tert-butyl 2-(cyanomethyl)piperazine-1-carboxylate (CAS: 1808997-73-0) in step d.
Example 267a:
m/z ES+ [M+H]+ 658.2; 1H NMR (400 MHz, CDCl3) δ ppm 8.34 (s, 1H), 7.50-7.43 (m, 1H), 7.35-7.29 (m, 2H), 7.06 (s, 1H), 6.63-6.43 (m, 1H), 6.37-6.26 (m, 1H), 5.75 (d, J=9.6 Hz, 1H), 5.07-4.88 (m, 0.5H), 4.63 (d, J=9.2 Hz, 1H), 3.85-3.69 (m, 0.5H), 3.61-3.51 (m, 1H), 3.39 (s, 3H), 3.34-3.09 (m, 2H), 3.08-2.94 (m, 3H), 2.93-2.71 (m, 5H), 2.68-2.55 (m, 1H), 2.49 (s, 3H), 2.47-2.40 (m, 2H), 2.26-2.19 (m, 2H), 2.10-1.98 (m, 1H), 1.54-1.47 (m, 2H).
Example 267b:
m/z ES+ [M+H]+ 658.2; 1H NMR (400 MHz, CDCl3) δ ppm 8.34 (s, 1H), 7.48-7.41 (m, 1H), 7.35-7.28 (m, 2H), 7.06 (s, 1H), 6.63-6.44 (m, 1H), 6.37-6.28 (m, 1H), 5.76 (d, J=11.2 Hz, 1H), 5.04-4.89 (m, 0.5H), 4.63 (d, J=9.2 Hz, 1H), 3.86-3.68 (m, 0.5H), 3.62-3.52 (m, 1H), 3.39 (s, 3H), 3.35-3.14 (m, 1H), 3.13-2.71 (m, 9H), 2.69-2.56 (m, 1H), 2.49-2.46 (m, 4H), 2.42-2.30 (m, 1H), 2.28-2.19 (m, 1H), 2.19-2.10 (m, 1H), 2.09-1.98 (m, 1H), 1.56-1.52 (m, 2H).
Step a. A mixture of Intermediate 9b (0.3 g, 0.60 mmol) and CDI (127 mg, 0.79 mmol) in NMP (5 mL) was stirred at 20° C. for 30 min, after which Intermediate F1 (208 mg, 0.91 mmol) was added. The reaction mixture was stirred at 120° C. for 2 h. Upon completion, the residue was diluted with water (30 mL) and extracted with EtOAc (3×30 mL). The combined organic layers were washed with brine (75 mL), dried over Na2SO4, evaporated and purified by column chromatography (PE/EtOAc=1/1) to give tert-butyl (R)-2-(5-(((3aR,11aS)-6-chloro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)methyl)-1,2,4-oxadiazol-3-yl)pyrrolidine-1-carboxylate (0.27 g, 65% yield) as a colourless oil.
m/z ES+ [M+H]+ 690.4; 1H NMR (400 MHz, CDCl3) δ ppm 8.31 (s, 1H), 7.48-7.43 (m, 1H), 7.35-7.31 (m, 2H), 7.03 (s, 1H), 5.30 (s, 1H), 5.08-4.89 (m, 1H), 4.87-4.67 (m, 1H), 4.66-4.58 (m, 1H), 3.76-3.63 (m, 1H), 3.63-3.45 (m, 2H), 3.35-3.24 (m, 4H), 3.16-3.08 (m, 1H), 2.73 (dd, J=16.8, 8.4 Hz, 1H), 2.46 (s, 3H), 2.25-2.19 (m, 2H), 1.96-1.78 (m, 3H), 1.45-1.41 (m, 9H).
Step b. This step was conducted in a similar manner to Example 77, step b.
Steps c-d. These 2 steps were conducted in a similar manner to Example 51, steps b-c.
m/z ES+ [M+H]+ 624.3; 1H NMR (400 MHz, CDCl3) δ ppm 8.34 (s, 1H), 7.27-7.20 (m, 3H), 7.05 (s, 1H), 6.56-6.31 (m, 2H), 5.76-5.55 (m, 1H), 5.40-5.11 (m, 1H), 4.75 (d, J=9.6 Hz, 1H), 4.48-4.36 (m, 1H), 4.32-4.13 (m, 1H), 3.91-3.60 (m, 3H), 3.44-3.33 (m, 3H), 3.14-2.99 (m, 1H), 2.97-2.84 (m, 1H), 2.82-2.71 (m, 1H), 2.49 (s, 3H), 2.41-2.36 (m, 3H), 2.34-2.14 (m, 3H), 2.13-1.99 (m, 2H).
The title compound was prepared in a similar manner to Example 268, using Intermediate 9b and Intermediate F2.
m/z ES+ [M+H]+ 624.3; 1H NMR (400 MHz, CDCl3) δ ppm 8.34 (s, 1H), 7.30-7.27 (m, 1H), 7.26-7.23 (m, 2H), 7.05 (s, 1H), 6.56-6.33 (m, 2H), 5.76-5.56 (m, 1H), 5.38-5.13 (m, 1H), 4.80-4.70 (m, 1H), 4.42 (d, J=16.0 Hz, 1H), 4.27-4.17 (m, 1H), 3.91-3.63 (m, 3H), 3.38 (s, 3H), 3.14-2.99 (m, 1H), 2.96-2.83 (m, 1H), 2.82-2.69 (m, 1H), 2.49 (s, 3H), 2.41-2.35 (m, 3H), 2.30-2.14 (m, 3H), 2.11-1.98 (m, 2H).
Step a. This step was conducted in a similar manner to Example 101, step a, using Intermediate 5b.
Step b. This step was conducted in a similar manner to Example 77, step b.
Steps c-g. These 5 steps were conducted in a similar manner to Example 101, steps b-f, using (tert-butoxycarbonyl)-D-proline in step d.
m/z ES+ [M+H]+ 624.3; 1H NMR (400 MHz, CDCl3) δ ppm 8.33 (s, 1H), 7.26-7.18 (m, 3H), 7.04 (s, 1H), 6.54-6.25 (m, 2H), 5.79-5.62 (m, 1H), 5.42-5.24 (m, 1H), 4.73 (d, J=9.6 Hz, 1H), 4.29-4.21 (m, 1H), 4.20-4.07 (m, 1H), 3.94-3.76 (m, 1H), 3.76-3.63 (m, 2H), 3.38-3.27 (m, 3H), 3.15-2.97 (m, 1H), 2.94-2.80 (m, 1H), 2.79-2.68 (m, 1H), 2.48 (s, 3H), 2.41-2.29 (m, 4H), 2.26-2.03 (m, 4H).
The title compound was prepared in a similar manner to Example 270, using (tert-butoxycarbonyl)-D-proline in step d.
m/z ES+ [M+H]+ 624.3; 1H NMR (400 MHz, CDCl3) δ ppm 8.33 (s, 1H), 7.26-7.19 (m, 3H), 7.04 (s, 1H), 6.57-6.21 (m, 2H), 5.80-5.59 (m, 1H), 5.40-5.21 (m, 1H), 4.77-4.69 (m, 1H), 4.30-4.21 (m, 1H), 4.19-4.08 (m, 1H), 3.96-3.78 (m, 1H), 3.76-3.64 (m, 2H), 3.38-3.27 (m, 3H), 3.14-2.95 (m, 1H), 2.91-2.80 (m, 1H), 2.78-2.68 (m, 1H), 2.49 (s, 3H), 2.41-2.29 (m, 4H), 2.27-2.05 (m, 4H).
Step a. This step was conducted in a similar manner to Example 103, step a, using Intermediate 9b and Intermediate G1.
Step b. To a solution of tert-butyl (R)-2-(2-(2-((3aR,11aS)-6-chloro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)acetyl)hydrazine-1-carbonyl)pyrrolidine-1-carboxylate (300 mg, 0.42 mmol) in DCM (10 mL) was added DIPEA (547 mg, 4.24 mmol) and Burgess reagent (CAS: 29684-56-8; 1.01 g, 4.24 mmol). The mixture was stirred at 25° C. for 12 h. Upon completion, the reaction mixture was diluted with water (15 mL) and extracted with DCM (3×20 mL). The combined organic layers were washed with brine (40 mL), dried over Na2SO4, evaporated and purified by column chromatography (EtOAc) to give tert-butyl (R)-2-(5-(((3aR,11aS)-6-chloro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)methyl)-1,3,4-oxadiazol-2-yl)pyrrolidine-1-carboxylate (180 mg, 57% yield) as a yellow oil.
m/z ES+ [M+H]+ 690.3.
Step c. This step was conducted in a similar manner to Example 77, step b.
Steps d-e. These 2 steps were conducted in a similar manner to Example 51, steps b-c.
m/z ES+ [M+H]+ 624.4; 1H NMR (400 MHz, CDCl3) δ ppm 8.34 (s, 1H), 7.27-7.19 (m, 3H), 7.10-6.97 (m, 1H), 6.54-6.43 (m, 1H), 6.43-6.35 (m, 1H), 5.91-5.58 (m, 1H), 5.44-5.25 (m, 1H), 4.71 (d, J=9.2 Hz, 1H), 4.47-4.05 (m, 2H), 3.92-3.59 (m, 3H), 3.45-3.21 (m, 3H), 3.10-2.84 (m, 2H), 2.81-2.68 (m, 1H), 2.48 (s, 3H), 2.38-2.33 (m, 3H), 2.32-2.02 (m, 5H).
The title compound was prepared in a similar manner to Example 272, using Intermediate 9b and Intermediate G2 in step a.
m/z ES+ [M+H]+ 624.2; 1H NMR (400 MHz, DMSO-d6) δ ppm 8.18 (s, 1H), 7.35 (s, 1H), 7.33-7.24 (m, 2H), 7.24-7.19 (m, 1H), 6.68-6.36 (m, 1H), 6.17-6.08 (m, 1H), 5.76-5.57 (m, 1H), 5.56-5.19 (m, 1H), 4.65 (d, J=9.6 Hz, 1H), 4.38-4.30 (m, 1H), 4.29-4.21 (m, 1H), 3.79-3.43 (m, 3H), 3.28-3.24 (m, 3H), 3.06-2.96 (m, 1H), 2.86-2.73 (m, 1H), 2.63-2.53 (m, 1H), 2.48 (s, 3H), 2.44-2.36 (m, 1H), 2.32-2.20 (m, 4H), 2.16-1.78 (m, 3H).
The title compound was prepared in a similar manner to Example 272, using Intermediate 9b and Intermediate G3 in step a.
m/z ES+ [M+H]+ 610.2; 1H NMR (400 MHz, CDCl3) δ ppm 8.34 (s, 1H), 7.25 (s, 3H), 7.05 (s, 1H), 6.42-6.30 (m, 1H), 6.28-6.04 (m, 1H), 5.82-5.53 (m, 2H), 4.72 (d, J=8.8 Hz, 1H), 4.54-4.13 (m, 4H), 3.76-3.62 (m, 1H), 3.48-3.31 (m, 3H), 2.98-2.62 (m, 5H), 2.50 (s, 3H), 2.38 (s, 3H), 2.26-2.14 (m, 1H).
The title compound was prepared in a similar manner to Example 272, using Intermediate 9b and Intermediate G4 in step a.
m/z ES+ [M+H]+ 610.2; 1H NMR (400 MHz, CDCl3) δ ppm 8.33 (s, 1H), 7.26-7.24 (m, 3H), 7.05 (s, 1H), 6.43-6.29 (m, 1H), 6.28-5.91 (m, 1H), 5.83-5.50 (m, 2H), 4.80-4.66 (m, 1H), 4.50-4.18 (m, 4H), 3.79-3.63 (m, 1H), 3.35 (s, 3H), 3.00-2.63 (m, 5H), 2.49 (s, 3H), 2.44-2.33 (m, 3H), 2.25-2.12 (m, 1H).
The Examples in Table 20 were prepared using methods similar to those described in the synthesis of Example 38 or Example 39, using the listed Intermediates in step a. For Examples 287a/b-294a/b an additional chiral SFC step was conducted after step b.
1H NMR (400 MHz) δ ppm
The Examples in Table 21 were prepared using methods similar to those described in the synthesis of Example 39 or Example 43, using the listed Intermediates in step a, and the appropriate warhead in step c. For Examples 310a/b-311a/b, an additional chiral SFC step was conducted after step a.
1H NMR (400 MHz) δ ppm
The Examples in Table 22 were prepared using methods similar to those described in the synthesis of Example 77, using the listed Intermediates in step a. The final step was conducted in a similar manner to the amide coupling with acryloyl chloride described for Example 1. For Examples 318a/b-324a/b, an additional chiral SFC step was conducted after step b.
1H NMR (400 MHz)
The Examples in Table 23 were prepared using methods similar to those described in the synthesis of Example 77, using the listed Intermediates in step a, and the appropriate warhead in step d. For Examples 329a/b-330a/b, an additional chiral SFC step was conducted after step a. For Examples 331-332, epimerisation occurred during step a, and the isomers were separated by chiral SFC after step b. The absolute configuration of Examples 331-332 was determined by 2D NMR.
1H NMR (400 MHz)
Polθ Polymerase Domain Kinact/KI Assay
A PicoGreen assay was used to measure the Kinact and KI values of covalent compounds that inhibit the activity of Polθ in vitro.
Human Polθ polymerase domain (aa1820-2590) was expressed in E. coli, purified, aliquoted and stored at −80° C. until required. Polθ substrate was generated from a 1.2:1 mix of DNA II Short to DNA II Long to give a final concentration of 20 mM substrate in annealing buffer (20 mM Tris pH 7.5, 50 mM NaCl). The substrate was heated in 50 mL aliquots to 95° C. in a heating block for 5 min before the heating block switched off, the reaction left to cool to rt and stored at −20° C. until required.
Assay measurements were performed with 1× buffer comprising 25 mM Tris pH 7.5, 12.5 mM NaCl, 0.5 mM NaCl, 5% (v/v) glycerol, 0.01% v/v Triton x-100, 0.1 mg/ml BSA, 1 mM DTT. Test compounds were prepared by dilution in 100% DMSO to give a 12 μM intermediate stock of each (100× final top concentration). 100 nL of 23×1:1.5-fold serial dilutions and a DMSO only control were dispensed using the Tecan dispenser into Greiner 384 well black low volume plates (product code 784076). DMSO concentration was maintained at 1% of the final assay volume by back filling with DMSO.
2× Working stock of substrate (200 nM of DNA substrate and 100 mM dNTPs) and enzyme (1.4 nM PolΘ) were made up in assay buffer. 5 μL/well of both enzyme and substrate 2× solutions were dispensed using a Tempest dispenser (Formulatrix) into assay plates that had been pre-dispensed with compound to give a final assay concentration of 100 nM DNA substrate, 50 mM dNTPs and 0.8 nM Polθ. To stop the reaction, 5 mL of a solution containing 25 mM Tris-HCl pH 7.5 and 20 mM EDTA was added at 6 timepoints (t=0, 2.5, 5, 7.5, 10, 12.5, 15, 20, 25, 30, 35, 40, 45, 50, 55 and 60 min) using the Tempest's time delay function. The plates were covered during the time course to prevent evaporation. After completing the assay, 5 μL of detection reagent (25 mM Tris-HCl pH 7.5 and 2.5% (v/v) PicoGreen) was dispensed into the wells using a Tempest liquid handler (Formulatrix) and plates subsequently read on the CLARIOstar Plus (BMG Labtech) using the default optical settings for fluorescein and the auto gain/focus settings.
All data analysis was carried out using GraphPad Prism V.8 (GraphPad Software Inc, San Diego, CA) Time course data for each inhibitor concentration was fitted to equation 1 to determine kobs values.1 Any time points where the control (DMSO alone) reaction was no longer linear were excluded from the analysis. 1) Evaluation of Enzyme Inhibitors in Drug Discovery (Ch9), R. A Copeland, Wiley, 2nd Edition 2013.
A secondary plot of kobs vs [I] is used to determine the Kinact and KI parameters (for a 2-step covalent inhibitor) by fitting to equation 2.1
The compounds of Examples 1 to 332 were tested in the above mentioned Polθ Polymerase Domain Kinact/KI Assay and the results are shown in the following table:
| Number | Date | Country | Kind |
|---|---|---|---|
| 2115158.4 | Oct 2021 | GB | national |
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/GB2022/052687 | 10/21/2022 | WO |
