Patent Application
20200246347
The present invention relates to protein-inhibitory substituted pyrrolopyridine derivatives, to pharmaceutical compositions and combinations comprising the compounds according to the invention, and to the prophylactic and therapeutic use of the inventive compounds, respectively to the use of said compounds for manufacturing pharmaceutical compositions for the treatment or prophylaxis of diseases, in particular for neoplastic disorders, respectively cancer or conditions with dysregulated immune responses or other disorders associated with aberrant MAP4K1 signaling, as a sole agent or in combination with other active ingredients. The present invention further relates to the use, respectively to the use of said compounds for manufacturing pharmaceutical compositions for the treatment or prophylaxis of protein inhibitors in benign hyperplasias, atherosclerotic disorders, sepsis, autoimmune disorders, vascular disorders, viral infections, in neurodegenerative disorders, in inflammatory disorders, in atherosclerotic disorders and in male fertility control.
Although cancer cell commonly can be recognize by the adaptive immune system, the response generated is evidently not capable of eliminating the tumor. A major reason for this is the presence of immunosuppressive mechanisms in the tumor microenvironment. In this respect, inhibitors of T-cell immune checkpoint such as CTLA-4, PD-1 or PD-L1 were recently shown to result in a remarkable clinical efficacy in subsets of cancer patients. Besides cell surface receptors that act as negative immune regulators, several mediators of intracellular signaling have been identified that also represent potential immunoevasive mechanisms utilized by the tumor.
One of these is MAP4K1, also known as hematopoietic progenitor kinase 1 (HPK1). MAP4K1 (GeneID11184) is a serine/threonine kinase and member of the Germinal Center Kinase family. In the adult organism MAP4K1 expression is restricted to hematopoietic cell types. The MAP4K1 protein consist of a N-terminal kinase domain, followed by a proline-rich domain that can interact with adaptor molecules through SH2 and SH3 domains, and a C-terminal citron homology domain of which the exact function remains to be identified. Through its proline-rich domain, MAP4K1 is capable of binding to a diversity of adaptors in hematopoietic cells, including those involved in T-cell receptor (TCR), B-cell receptor (BCR) and cytokine signaling (Hu et al., Genes Dev. 1996 Sep. 15; 10(18):2251-64, 2; Ling et al., J Biol Chem. 2001 June 1; 276(22), Sauer et al., J Biol Chem. 2001 Nov. 30; 276(48):45207-16., Tsuji et al., J Exp Med. 2001 Aug. 20; 194(4):529-39, Boomer et al., J Cell Biochem. 2005 May 1; 95(1):34-44). The function of MAP4K1 has been studied in greatest detail in the context of TCR signaling. Upon TCR stimulation, MAP4K1 is phosphorylated on tyrosine 381 (Y-381; Y-379 in mouse) (Di Bartolo et al., J Exp Med. 2007 Mar. 19; 204(3):681-91). Consequently, MAP4K1 is recruited to the TCR-signaling complex where it induces dissociation of this complex through its serine/threonine kinase function. In particular MAP4K1 phosphorylates the SLP-76 adaptor protein at Serine-376, resulting in downregulation of AP-1 and Erk2 pathways. As, such, MAP4K1 acts as a negative feedback on TCR-signaling (Liou et al., Immunity. 2000 April; 12(4):399-408; Lasserre et al., J Cell Biol. 2011 Nov. 28; 195(5):839-53). Alternatively, MAP4K1 can be triggered to suppress T cell function by prostaglandin E2 (PGE2), and possibly also by transforming growth factor beta (TGF-beta), factors that are commonly found in the tumor microenvironment. Notably, MAP4K1 activation by these mediators involves protein kinase A (PKA)-dependent phosphorylation of Serine 171 (S-171; also in mouse) (Alzabin et al., Cancer Immunol Immunother. 2010 March; 59(3):419-29; Sawasdikosol et al., J Biol Chem. 2007 Nov. 30; 282(48):34693-9).
Further important insights into the function of MAP4K1 in the regulation of T cell immunity stem from in vivo and in vitro experiments respectively with MAP4K1 deficient mice produced by two laboratories and with immune cells isolated from these mice (Shui et al., Nat Immunol. 2007 January; 8(1):84-91; Alzabin et al., Cancer Immunol Immunother. 2010 March; 59(3):419-29). MAP4K1-deficient mice show an apparent normal phenotype, are fertile and exhibit normal lymphocyte development. These animals are prone to develop T-cell dependent autoimmune reactivity as indicated by development of a more severe disease score in the EAE (experimental autoimmune encephalomyelitis) model of multiple sclerosis (Shui et al., Nat Immunol. 2007 January; 8(1):84-91). In case of the second strain, a dysregulation of immune function was observed when, at the age of approximately 6 months, MAP4K1-deficient mice develop a spontaneous autoimmune phenotype (Alzabin et al., Cancer Immunol Immunother. 2010 March; 59(3):419-29). In vitro studies showed that MAP4K1−/−T-cells display hyper-responsiveness upon TCR-stimulation. These cells proliferate and secrete pro-inflammatory cytokines like IL-2 or IFNg to a significantly greater extent than their wild-type counterparts (Shui et al., Nat Immunol. 2007 January; 8(1):84-91). Furthermore, MAP4K1−/− T-cells are resistant to PGE2-mediated suppression of T cell proliferation, suppression of IL-2 production and induction of apoptosis (Alzabin et al., Cancer Immunol Immunother. 2010 March; 59(3):419-29). In the context of tumor immunology, in vivo experiments revealed that MAP4K1−/− mice are much more resistant to tumorigenesis by PGE2-producing Lewis lung carcinoma than wild type mice, which correlated with increased T-lymphocyte infiltration in the tumor areas. The crucial role of T-cells in tumor rejection was supported by experiments in which MAP4K1−/− T-cells adoptively transferred into T-cell-deficient mice were able to eradicate tumors more efficiently than wild-type T-cells (Alzabin et al., Cancer Immunol Immunother. 2010 March; 59(3):419-29). The important role of the kinase enzymatic activity was demonstrated by studies were only wild type MAP4K1, but not the MAP4K1 kinase-dead mutant, could mediate serine-phosphorylation of the TCR-signaling complex component SLP-76 and subsequent binding of SLP-76 to the negative regulator of TCR-signaling 14-3-3-t (Shui et al., Nat Immunol. 2007 January; 8(1):84-91). MAP4K1 also regulates the stimulation and activation of dendritic cells. MAP4K1 deficient Bone marrow derived cells (BMDC) express after maturation and stimulation higher level of costimulatory molecules and produce more proinflammatory cytokines. Also elimination of tumors was observed to be more efficient by MAP4K1−/−BMDC compared to their wildtype counterparts (Alzabin et al., J Immunol. 2009 May 15; 182(10):6187-94).
In WO 2016/205942 HPK1, respectively inhibitors and methods of their use in cancer treatment are described. Especially, the application concerns thieno-pyridinones that can be used in anti-cancer therapy. These compounds differ from the instant compounds in their chemical structure.
In WO 2016/195776 inhibitors and methods for leukemia, cancer and diabetes treatment dependent on inhibition the interaction of menin with of MLL1, MLL2 and MLL-fusion oncoproteins are described. These compounds differ from the instant compounds in their chemical structure.
In WO 2006/014325 C-MET modulators and their use in cancer treatment are described. These compounds differ from the instant compounds in their chemical structur.
In WO 2005/058891 Rho kinase inhibitors and their use in cardiovascular and cancer treatment are described. These compounds differ from the instant compounds in their chemical structure.
In WO 2015/089479 several inhibitors are described that show inhibition of several kinases (e.g., BTK, HCK, TAK1 and HPK1). These compounds differ from the instant compounds in their chemical structure.
It would therefore be desirable to provide novel compounds having prophylactic and therapeutic properties.
Accordingly, it is an object of the present invention to provide compounds and pharmaceutical compositions comprising these compounds used for prophylactic and therapeutic applications for hyperproliferative disorders, in particular for cancer, respectively tumour disorders, and conditions with dysregulated immune responses, as a sole agent or in combination with other active ingredients.
A further object of the present invention is to provide compounds and pharmaceutical compositions comprising these compounds for manufacturing pharmaceutical compositions for the treatment or prophylaxis of protein inhibitors in benign hyperplasias, atherosclerotic disorders, sepsis, autoimmune disorders, vascular disorders, viral infections, in neurodegenerative disorders, in inflammatory disorders and in male fertility control.
Surprisingly, the compounds according to the invention inhibit the MAP4K1 protein and inhibit the growth of cancer cells. Accordingly, they provide novel structures for the therapy of human and animal disorders, in particular of cancers.
The present invention relates to compounds of formula (I)
in which
The compounds of formula (I) are particularly suitable for a large number of prophylactic and therapeutic applications, in particular for hyperproliferative disorders, for tumour disorders and as protein inhibitors and further for viral infections, for neurodegenerative disorders, for inflammatory disorders, for atherosclerotic disorders and for male fertility control.
Further, it covers their use in combination with other anti cancer medications such as immunotherapeutics, targeted anti cancer agents, radiation or chemotherapy.
The term “substituted” means that one or more hydrogen atoms on the designated atom or group are replaced with a selection from the indicated group, provided that the designated atom's normal valency under the existing circumstances is not exceeded. Combinations of substituents and/or variables are permissible.
The term “optionally substituted” means that the number of substituents can be equal to or different from zero. Unless otherwise indicated, it is possible that optionally substituted groups are substituted with as many optional substituents as can be accommodated by replacing a hydrogen atom with a non-hydrogen substituent on any available carbon atom. Commonly, it is possible for the number of optional substituents, when present, to be 1, 2 or 3.
The term “comprising” when used in the specification includes “consisting of”.
If within the present text any item is referred to as “as mentioned herein”, it means that it may be mentioned anywhere in the present text.
The terms as mentioned in the present text have the following meanings:
The term “halogen” means a fluorine, chlorine, bromine or iodine, particularly a fluorine, chlorine or bromine atom.
The term “C1-C6-alkyl” means a linear or branched, saturated, monovalent hydrocarbon group having 1, 2, 3, 4, 5 or 6 carbon atoms, e.g. a methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, isobutyl, tert-butyl, pentyl, isopentyl, 2-methylbutyl, 1-methylbutyl, 1-ethylpropyl, 1,2-dimethylpropyl, neo-pentyl, 1,1-dimethylpropyl, hexyl, 1-methylpentyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 1-ethylbutyl, 2-ethylbutyl, 1,1-dimethylbutyl, 2,2-dimethylbutyl, 3,3-dimethylbutyl, 2,3-dimethylbutyl, 1,2-dimethylbutyl or 1,3-dimethylbutyl group, or an isomer thereof. Particularly, said group has 1, 2, 3 or 4 carbon atoms (“C1-C4-alkyl”), e.g. a methyl, ethyl, propyl, isopropyl, butyl, sec-butyl isobutyl, or tert-butyl group, more particularly 1, 2 or 3 carbon atoms (“C1-C3-alkyl”), e.g. a methyl, ethyl, n-propyl or isopropyl group.
The term “hydroxy-C1-C6-alkyl” means a linear or branched, saturated, monovalent hydrocarbon group in which the term “C1-C6-alkyl” is defined supra, and in which 1 or 2 hydrogen atoms are replaced with a hydroxy group, e.g. a hydroxymethyl, 1-hydroxyethyl, 2-hydroxyethyl, 1,2-dihydroxyethyl, 3-hydroxypropyl, 2-hydroxypropyl, 1-hydroxypropyl, 1-hydroxypropan-2-yl, 2-hydroxypropan-2-yl, 2,3-dihydroxypropyl, 1,3-dihydroxypropan-2-yl, 3-hydroxy-2-methyl-propyl, 2-hydroxy-2-methyl-propyl, 1-hydroxy-2-methyl-propyl group.
The term “C1-C6-alkoxy” means a linear or branched, saturated, monovalent group of formula (C1-C6-alkyl)-O—, which means methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, sec-butoxy, isobutoxy or tert-butoxy.
The term “C1-C6-alkoxy-C1-C6-alkyl-” means a linear or branched, saturated, monovalent group of formula (C1-C6-alkyl)-O—(C1-C6-alkyl)-, in which the term “C1-C6-alkyl” is defined supra, and in which 1 to 3 hydrogen atoms of the C1-C6-alkyl group are replace by C1-C6-alkoxy.
The term “C1-C6-alkylamino-” means an amino radical having one or two alkyl substituents (selected independently of one another) having generally 1 to 6 (C1-C6-alkylamino) and preferably 1 to 3 (C1-C3-alkylamino) carbon atoms. (C1-C3)-Alkylamino represents, for example, a monoalkylamino radical having 1 to 3 carbon atoms or a dialkylamino radical having 1 to 3 carbon atoms each per alkyl substituent.
The following may be mentioned by way of example:
methylamino, ethylamino, n-propylamino, isopropylamino, tert-butylamino, n-pentylamino, n-hexylamino, N,N-dimethylamino, N,N-diethylamino, N-ethyl-N-methylamino, N-methyl-N-n-propylamino, N-isopropyl-N-n-propylamino, N-tert-butyl-N-methylamino, N-ethyl-N-n-pentylamino and N-n-hexyl-N-methylamino.
The term “amino-C1-C6-alkyl-” means a linear or branched, saturated, monovalent hydrocarbon group in which the term “C1-C6-alkyl” is defined supra, and in which 1 or 2 hydrogen atoms are replaced with an amino group, e.g. a aminomethyl, 1-aminoethyl, 2-aminoethyl, 1,2-diaminoethyl, 3-aminopropyl, 2-aminopropyl, 1-aminopropyl, 1-aminopropan-2-yl, 2-aminopropan-2-yl, 2,3-diaminopropyl, 1,3-diaminopropan-2-yl, 3-amino-2-methyl-propyl, 2-amino-2-methyl-propyl, 1-amino-2-methyl-propyl group.
The term “C1-C6-alkylamino-C1-C6-alkyl means that the alkylaminoalkyl group is attached via the alkyl moiety to the remainder of the molecule, e.g. N,N-dimethylaminoethyl-, N,N-dimethylaminomethyl-, N,N-diethylaminoethyl-, N,N-dimethylaminopropyl-, N-methylaminoethyl-, N-methylaminomethyl-.
The term “halo-C1-C6-alkyl-” means an alkyl radical having at least one halogen substituent. The term “C1-C6-alkyl-” is as defined supra.
A halo-C1-C6-alkyl radical is an alkyl radical having 1-6 carbon atoms and at least one halogen substituent. If a plurality of halogen substituents is present, these may also be different from one another. Preference is given to fluoro-C1-C6-alkyl, fluoro-C1-C4-alkyl, fluoro-C1-C3-alkyl, chloro-C1-C6-alkyl, chloro-C1-C4-alkyl, chloro-C1-C3-alkyl, bromo-C1-C6-alkyl, bromo-C1-C4-alkyl and bromo-C1-C3-alkyl radicals.
In this regard preferred are difluoromethyl, trifluoromethyl, 2,2,2-trifluoroethyl, pentafluoroethyl, 4,4,5,5,5-pentafluoropentyl or 3,3,4,4,5,5,5-heptafluoropentyl.
Preference is given to perfluorinated alkyl radicals such as trifluoromethyl or 2,2,2-trifluoroethyl.
The term “haloalkoxy” means an alkoxy radical having at least one halogen substituent. A halo-C1-C6-alkoxy radical is an alkoxy radical having 1-6 carbon atoms and at least one halogen substituent. If a plurality of halogen substituents is present, these may also be different from one another. Preference is given to fluoro-C1-C6-alkoxy, fluoro-C1-C4-alkoxy, fluoro-C1-C3-alkoxy, chloro-C1-C6-alkoxy, chloro-C1-C4-alkoxy, chloro-C1-C3-alkoxy, bromo-C1-C6-alkoxy, bromo-C1-C4-alkoxy and bromo-C1-C3-alkoxy radicals.
In this regard preferred are trifluoromethoxy and 2,2,2-trifluoroethoxy radicals.
The term “halophenyl” means a phenyl radical which is mono- or polysubstituted by identical or different substituents from the group consisting of fluorine, chlorine and bromine.
The term “C3-C10-cycloalkyl” means a saturated, monovalent, monocyclic hydrocarbon ring which contains 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms (“C3-C10-cycloalkyl”). Said C3-C10-cycloalkyl group is a monocyclic hydrocarbon ring, e.g. a cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl or cyclodecanyl. Preferred are 3 to 8 (C3-C8-cycloalkyl) and particularly preferably 3 to 7 (C3-C7-cycloalkyl) carbon atoms.
The term “C1-C6-alkylcarbonylamino” means the group alkyl-C(═O)—NH—having generally 1 to 6 (C1-C6-alkylcarbonylamino), preferably 1 to 4 and particularly preferably 1 to 3 carbon atoms in the alkyl moiety.
The term “phenyl-C1-C6-alkyl-” is understood to mean a group composed of an optionally substituted phenyl radical and a C1-C6-alkyl group, and bonded to the rest of the molecule via the C1-C6-alkyl group. Here, the C1-C6-alkyl is as defined supra.
Examples which may be mentioned include benzyl, phenethyl, phenylpropyl, phenylpentyl, with benzyl being preferred.
The term “monocyclic heterocyclyl-” means a non-aromatic monocyclic ring system having one, two or three heteroatoms which may be identical or different. The heteroatoms may be nitrogen atoms, oxygen atoms or sulphur atoms. The sulphur atom may be substituted with oxygene to form a —S(═O)— or —S(═O)2— group in the ring.
A monocyclic heterocyclyl ring according to the present invention may have 3 to 8, preferably 4 to 7, particularly preferably 5 or 6 ring atoms.
By way of example and with preference, the following may be mentioned for monocyclic heterocyclyl radicals having 3 ring atoms:
aziridinyl-.
By way of example and with preference, the following may be mentioned for monocyclic heterocyclyl radicals having 4 ring atoms:
azetidinyl-, oxetanyl- and 1,1-dioxidothietanyl-.
By way of example and with preference, the following may be mentioned for monocyclic heterocyclyl radicals having 5 ring atoms:
pyrrolidinyl-, imidazolidinyl-, pyrazolidinyl-, pyrrolinyl-, dioxolanyl-, 1,1-dioxidotetrahydro-thiophen-yl-, oxopyrrolidinyl- and tetrahydrofuranyl-.
By way of example and with preference, the following may be mentioned for monocyclic heterocyclyl radicals having 6 ring atoms:
piperidinyl-, piperazinyl-, morpholinyl-, dioxanyl-, tetrahydropyranyl-, 1,1-dioxidotetrahydro-2H-thiopyranyl-, oxopiperidinyl- and thiomorpholinyl-.
By way of example and with preference, the following may be mentioned for monocyclic heterocyclyl radicals having 7 ring atoms:
azepanyl-, oxepanyl-, 1,3-diazepanyl-, 1,4-diazepanyl-.
By way of example and with preference, the following may be mentioned for monocyclic heterocyclyl radicals having 8 ring atoms:
oxocanyl-, azocanyl-.
From among the monocyclic heterocyclyl radicals, preference is given to 4- to 7-membered saturated heterocyclyl radicals having up to two heteroatoms from the group consisting of O, N and S.
Particular preference is given to morpholinyl-, piperidinyl- and pyrrolidinyl-.
The term “4- to 10-membered heterocycloalkyl” means a monocyclic, saturated heterocycle with 4, 5, 6, 7, 8, 9 or 10 ring atoms in total, which contains one or two identical or different ring heteroatoms from the series N and O, it being possible for said heterocycloalkyl group to be attached to the rest of the molecule via any one of the carbon atoms or, if present, a nitrogen atom. The term “heterocycloalkyl” is as defined supra.
Said heterocycloalkyl group, without being limited thereto, can be a 4-membered ring, such as azetidinyl or oxetanyl, for example; or a 5-membered ring, such as tetrahydrofuranyl, 1,3-dioxolanyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl, 1,2-oxazolidinyl or 1,3-oxazolidinyl, for example; or a 6-membered ring, such as tetrahydropyranyl, piperidinyl, morpholinyl, piperazinyl, 1,3-dioxanyl, 1,4-dioxanyl or 1,2-oxazinanyl, for example.
Particularly, “4- to 6-membered heterocycloalkyl” means a 4- to 6-membered heterocycloalkyl as defined supra containing one ring oxygen atom and optionally one further ring heteroatom from the series: N, O. More particularly, “5- or 6-membered heterocycloalkyl” means a monocyclic, saturated heterocycle with 5 or 6 ring atoms in total, containing one ring oxygen atom.
The term “monocyclic heteroaryl” means a monovalent, aromatic ring having 5 or 6 ring atoms (a “5- or 6-membered heteroaryl” group), which contains at least one ring heteroatom and optionally one or two further ring heteroatoms from the series: N, O and/or S, and which is bound via a ring carbon atom or optionally via a ring nitrogen atom (if allowed by valency). Said heteroaryl group can be a 5-membered heteroaryl group, such as, for example, thienyl, furanyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl, thiadiazolyl or tetrazolyl; or a 6-membered heteroaryl group, such as, for example, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl or triazinyl.
In general, and unless otherwise mentioned, the heteroaryl or heteroarylene groups include all possible isomeric forms thereof, e.g.: tautomers and positional isomers with respect to the point of linkage to the rest of the molecule. Thus, for some illustrative non-restricting examples, the term pyridinyl includes pyridin-2-yl, pyridin-3-yl and pyridin-4-yl; or the term thienyl includes thien-2-yl and thien-3-yl.
Particularly, the heteroaryl group is a isothiazolyl, pyrazolyl, pyridinyl, pyridazinyl or pyrimidinyl group.
The terms “spirocycloalkyl” and “heterospirocycloalkyl” mean C5-C12-spirocycloalkyl or C5-C12-heterospirocycloalkyl where one, two, three or four carbon atoms are replaced by heteroatoms as defined above in any combination is understood to mean a fusion of two saturated ring systems which share one common atom.
Examples are spiro[2.2]pentyl, spiro[2.3]hexyl, azaspiro[2.3]hexyl, spiro[3.3]heptyl, azaspiro[3.3]heptyl, oxaazaspiro[3.3]heptyl, thiaazaspiro[3.3]heptyl, oxaspiro[3.3]heptyl, oxazaspiro[3.5]nonyl, oxazaspiro[3.4]octyl, oxazaspiro[5.5]undecyl, diazaspiro[3.3]heptyl, thiazaspiro[3.3]heptyl, thiazaspiro[3.4]octyl, azaspiro[5.5]decyl, and the further homologous spiro[3.4], spiro[4.4], spiro[5.5], spiro[6.6], spiro[2.4], spiro[2.5], spiro[2.6], spiro[3.5], spiro[3.6], spiro[4.5], spiro[4.6] and spiro[5.6] systems including the variants modified by heteroatoms as per the definition. Preference is given to C6-C10-heterospirocycloalkyl-, by way of example and with particular preference 2-azaspiro[3.3]heptyl-, 2,2-dioxido-2-thia-6-azaspiro[3.3]heptyl. 1,1-dioxido-1-thia-6-azaspiro[3.3]heptyl, 1-thia-6-azaspiro[3.3]heptyl-, 2-thia-6-azaspiro[3.3]heptyl-, 2-oxa-6-azaspiro[3.3]heptyl-,2,6-diazaspiro[3.3]heptyl-, 2-oxa-6-azaspiro[3.4]octyl-, 2-oxa-6-azaspiro[3.5]nonyl-,2-oxa-7-azaspiro[3.5]nonyl-, 8-azaspiro[4.5]decyl-, 2,8-diazaspiro[4.5]decyl- or 3-oxa-1,8-diazaspiro[4.5]decyl-.
The terms “bicycloalkyl” and “heterobicycloalkyl” mean C6-C12-bicycloalkyl or C6-C12-heterobicycloalkyl where one, two, three or four carbon atoms are replaced by heteroatoms as defined above in any combination is understood to mean a fusion of two saturated ring systems which share two directly adjacent atoms.
Examples are radicals derived from bicyclo[2.2.0]hexyl-, bicyclo[3.3.0]octyl-, bicyclo[4.4.0]decyl-, bicyclo[5.4.0]undecyl-, bicyclo[3.2.0]heptyl-, bicyclo[4.2.0]octyl-, bicyclo[5.2.0]nonyl-, bicyclo[6.2.0]decyl-, bicyclo[4.3.0]nonyl-, bicyclo[5.3.0]decyl-, bicyclo[6.3.0]undecyl- and bicyclo[5.4.0]undecyl-, including the variants modified by heteroatoms, for example azabicyclo[3.3.0]octyl-, azabicyclo[4.3.0]nonyl-, diazabicyclo[4.3.0]nonyl-, oxazabicyclo[4.3.0]nonyl-, thiazabicyclo[4.3.0]nonyl- or azabicyclo[4.4.0]decyl-, and the further possible combinations as per the definition. Preference is given to C6-C10-heterobicycloalkyl-, by way of example and with particular preference perhydrocyclopenta[c]pyrrolyl-, perhydrofuro[3,2-c]pyridinyl-, perhydropyrrolo[1,2-a]pyrazinyl-, perhydropyrrolo[3,4-c]pyrrolyl-.
Preferred examples of C6-C12-bicycloalkyl- are perhydronaphthalenyl- (decalinyl-), perhydrobenzoannulenyl-, perhydroazulenyl-, perhydroindanyl-, perhydropentalenyl-.
The terms “bridged cycloalkyl” and “bridged heterocycloalkyl” mean a bridged C6-C12 ring system such as bridged C6-C12-cycloalkyl- or bridged C6-C12-heterocycloalkyl- is understood to mean a fusion of at least two saturated rings which share two atoms that are not directly adjacent to one another. This may give rise either to a bridged carbocycle (bridged cycloalkyl-) or to a bridged heterocycle (bridged heterocycloalkyl-) where one, two, three or four carbon atoms are replaced by heteroatoms as defined above in any combination.
Examples are bicyclo[2.2.1]heptyl-, azabicyclo[2.2.1]heptyl-, oxazabicyclo[2.2.1]heptyl-, thiazabicyclo[2.2.1]heptyl-, diazabicyclo[2.2.1]heptyl-, bicyclo[2.2.2]octyl-, azabicyclo[2.2.2]octyl-, diazabicyclo[2.2.2]octyl-, oxazabicyclo[2.2.2]octyl-, thiazabicyclo[2.2.2]octyl-, bicyclo[3.2.1]octyl-, azabicyclo[3.2.1]octyl-, diazabicyclo[3.2.1]octyl-, oxazabicyclo[3.2.1]octyl-, thiazabicyclo[3.2.1]octyl-, bicyclo[3.3.1]nonyl-, azabicyclo[3.3.1]nonyl-, diazabicyclo[3.3.1]nonyl-oxazabicyclo[3.3.1]nonyl-, thiazabicyclo[3.3.1]nonyl-, bicyclo[4.2.1]nonyl-, azabicyclo[4.2.1]nonyl-, diazabicyclo[4.2.1]nonyl-, oxazabicyclo[4.2.1]nonyl-, thiazabicyclo[4.2.1]nonyl-, bicyclo[3.3.2]decyl-, azabicyclo[3.3.2]decyl-, diazabicyclo[3.3.2]decyl-, oxazabicyclo[3.3.2]decyl-, thiazabicyclo[3.3.2]decyl- or azabicyclo[4.2.2]decyl- and the further possible combinations according to the definition. Preference is given to bridged C6-C10-heterocycloalkyl-, by way of example and with particular preference 2-azabicyclo[2.2.1]heptyl-, 2,5-diazabicyclo[2.2.1]heptyl-, 2-oxa-5-azabicyclo[2.2.1]heptyl-, 8-azabicyclo[3.2.1]octyl-, 8-oxa-3-azabicyclo[3.2.1]octyl-, 3,9-diazabicyclo[4.2.1]nonyl-.
Where the plural form of the word compounds, salts, polymorphs, hydrates, solvates and the like, is used herein, this is taken to mean also a single compound, salt, polymorph, isomer, hydrate, solvate or the like.
By “stable compound’ or “stable structure” is meant a compound that is sufficiently robust to survive isolation to a useful degree of purity from a reaction mixture, and formulation into an efficacious therapeutic agent.
The compounds of the present invention optionally contain one or more asymmetric centres, depending upon the location and nature of the various substituents desired. It is possible that one or more asymmetric carbon atoms are present in the (R) or (S) configuration, which can result in racemic mixtures in the case of a single asymmetric centre, and in diastereomeric mixtures in the case of multiple asymmetric centres. In certain instances, it is possible that asymmetry also be present due to restricted rotation about a given bond, for example, the central bond adjoining two substituted aromatic rings of the specified compounds. Preferred compounds are those which produce the more desirable biological activity. Separated, pure or partially purified isomers and stereoisomers or racemic or diastereomeric mixtures of the compounds of the present invention are also included within the scope of the present invention. The purification and the separation of such materials can be accomplished by standard techniques known in the art.
Preferred isomers are those which produce the more desirable biological activity. These separated, pure or partially purified isomers or racemic mixtures of the compounds of this invention are also included within the scope of the present invention. The purification and the separation of such materials can be accomplished by standard techniques known in the art.
The optical isomers can be obtained by resolution of the racemic mixtures according to conventional processes, for example, by the formation of diastereoisomeric salts using an optically active acid or base or formation of covalent diastereomers. Examples of appropriate acids are tartaric, diacetyltartaric, ditoluoyltartaric and camphorsulfonic acid. Mixtures of diastereoisomers can be separated into their individual diastereomers on the basis of their physical and/or chemical differences by methods known in the art, for example, by chromatography or fractional crystallisation. The optically active bases or acids are then liberated from the separated diastereomeric salts. A different process for separation of optical isomers involves the use of chiral chromatography (e.g., HPLC columns using a chiral phase), with or without conventional derivatisation, optimally chosen to maximise the separation of the enantiomers. Suitable HPLC columns using a chiral phase are commercially available, such as those manufactured by Daicel, e.g., Chiracel OD and Chiracel OJ, for example, among many others, which are all routinely selectable. Enzymatic separations, with or without derivatisation, are also useful. The optically active compounds of the present invention can likewise be obtained by chiral syntheses utilizing optically active starting materials.
In order to distinguish different types of isomers from each other reference is made to IUPAC Rules Section E (Pure Appl Chem 45, 11-30, 1976).
The present invention includes all possible stereoisomers of the compounds of the present invention as single stereoisomers, or as any mixture of said stereoisomers, e.g. (R)- or (S)-isomers, in any ratio. Isolation of a single stereoisomer, e.g. a single enantiomer or a single diastereomer, of a compound of the present invention is achieved by any suitable state of the art method, such as chromatography, especially chiral chromatography, for example.
Further, the compounds of the present invention can exist as N-oxides, which are defined in that at least one nitrogen of the compounds of the present invention is oxidised. The present invention includes all such possible N-oxides.
The present invention also covers useful forms of the compounds of the present invention, such as metabolites, hydrates, solvates, prodrugs, salts, in particular pharmaceutically acceptable salts, and/or co-precipitates.
The compounds of the present invention can exist as a hydrate, or as a solvate, wherein the compounds of the present invention contain polar solvents, in particular water, methanol or ethanol for example, as structural element of the crystal lattice of the compounds. It is possible for the amount of polar solvents, in particular water, to exist in a stoichiometric or non-stoichiometric ratio. In the case of stoichiometric solvates, e.g. a hydrate, hemi-, (semi-), mono-, sesqui-, di-, tri-, tetra-, penta- etc. solvates or hydrates, respectively, are possible. The present invention includes all such hydrates or solvates.
Further, it is possible for the compounds of the present invention to exist in free form, e.g. as a free base, or as a free acid, or as a zwitterion, or to exist in the form of a salt. Said salt may be any salt, either an organic or inorganic addition salt, particularly any pharmaceutically acceptable organic or inorganic addition salt, which is customarily used in pharmacy, or which is used, for example, for isolating or purifying the compounds of the present invention.
The term “pharmaceutically acceptable salt” refers to an inorganic or organic acid addition salt of a compound of the present invention. For example, see S. M. Berge, et al. “Pharmaceutical Salts,” J. Pharm. Sci. 1977, 66, 1-19.
A suitable pharmaceutically acceptable salt of the compounds of the present invention may be, for example, an acid-addition salt of a compound of the present invention bearing a nitrogen atom, in a chain or in a ring, for example, which is sufficiently basic, such as an acid-addition salt with an inorganic acid, or “mineral acid”, such as hydrochloric, hydrobromic, hydroiodic, sulfuric, sulfamic, bisulfuric, phosphoric, or nitric acid, for example, or with an organic acid, such as formic, acetic, acetoacetic, pyruvic, trifluoroacetic, propionic, butyric, hexanoic, heptanoic, undecanoic, lauric, benzoic, salicylic, 2-(4-hydroxybenzoyl)-benzoic, camphoric, cinnamic, cyclopentanepropionic, digluconic, 3-hydroxy-2-naphthoic, nicotinic, pamoic, pectinic, 3-phenylpropionic, pivalic, 2-hydroxyethanesulfonic, itaconic, trifluoromethanesulfonic, dodecylsulfuric, ethanesulfonic, benzenesulfonic, para-toluenesulfonic, methanesulfonic, 2-naphthalenesulfonic, naphthalinedisulfonic, camphorsulfonic acid, citric, tartaric, stearic, lactic, oxalic, malonic, succinic, malic, adipic, alginic, maleic, fumaric, D-gluconic, mandelic, ascorbic, glucoheptanoic, glycerophosphoric, aspartic, sulfosalicylic, or thiocyanic acid, for example.
Further, another suitably pharmaceutically acceptable salt of a compound of the present invention which is sufficiently acidic, is an alkali metal salt, for example a sodium or potassium salt, an alkaline earth metal salt, for example a calcium, magnesium or strontium salt, or an aluminium or a zinc salt, or an ammonium salt derived from ammonia or from an organic primary, secondary or tertiary amine having 1 to 20 carbon atoms, such as ethylamine, diethylamine, triethylamine, ethyldiisopropylamine, monoethanolamine, diethanolamine, triethanolamine, dicyclohexylamine, dimethylaminoethanol, diethylaminoethanol, tris(hydroxymethyl)aminomethane, procaine, dibenzylamine, N-methylmorpholine, arginine, lysine, 1,2-ethylenediamine, N-methylpiperidine, N-methyl-glucamine, N,N-dimethyl-glucamine, N-ethyl-glucamine, 1,6-hexanediamine, glucosamine, sarcosine, serinol, 2-amino-1,3-propanediol, 3-amino-1,2-propanediol, 4-amino-1,2,3-butanetriol, or a salt with a quanternary ammonium ion having 1 to 20 carbon atoms, such as tetramethylammonium, tetraethylammonium, tetra(n-propyl)ammonium, tetra(n-butyl)ammonium, N-benzyl-N,N,N-trimethylammonium, choline or benzalkonium.
Those skilled in the art will further recognise that it is possible for acid addition salts of the claimed compounds to be prepared by reaction of the compounds with the appropriate inorganic or organic acid via any of a number of known methods. Alternatively, alkali and alkaline earth metal salts of acidic compounds of the present invention are prepared by reacting the compounds of the present invention with the appropriate base via a variety of known methods.
The present invention includes all possible salts of the compounds of the present invention as single salts, or as any mixture of said salts, in any ratio.
In the present text, in particular in the Experimental Section, for the synthesis of intermediates and of examples of the present invention, when a compound is mentioned as a salt form with the corresponding base or acid, the exact stoichiometric composition of said salt form, as obtained by the respective preparation and/or purification process, is, in most cases, unknown. Unless specified otherwise, suffixes to chemical names or structural formulae relating to salts, such as “hydrochloride”, “trifluoroacetate”, “sodium salt”, or “x HCl”, “x CF3COOH”, “x Na+”, for example, mean a salt form, the stoichiometry of which salt form not being specified.
This applies analogously to cases in which synthesis intermediates or example compounds or salts thereof have been obtained, by the preparation and/or purification processes described, as solvates, such as hydrates, with (if defined) unknown stoichiometric composition. Furthermore, the present invention includes all possible crystalline forms, or polymorphs, of the compounds of the present invention, either as single polymorph, or as a mixture of more than one polymorph, in any ratio.
Moreover, the present invention also includes prodrugs of the compounds according to the invention. The term “prodrugs” here designates compounds which themselves can be biologically active or inactive, but are converted (for example metabolically or hydrolytically) into compounds according to the invention during their residence time in the body. The invention further includes all possible crystallized and polymorphic forms of the inventive compounds, whereby the polymorphs are existing either as a single polymorph form or are existing as a mixture of several polymorphs in all concentrations.
The invention further includes all possible cyclodextrin clathrates, i.e alpha-, beta-, or gamma-cyclodextrins, hydroxypropyl-beta-cyclodextrins, methylbetacyclodextrins.
Of selected interest are those compounds of formula (I),
in which
Of selected interest are those compounds of formula (I),
in which
Of selected interest are those compounds of formula (I),
in which
Of selected interest are those compounds of formula (I),
in which
Of selected interest are those compounds of formula (I),
in which
and their polymorphs, enantiomers, diastereomers, racemates, tautomers, solvates, physiologically acceptable salts and solvates of these salts.
In accordance with a further embodiment, the present invention covers compounds of general formula (I), in which X represents an oxygene atom.
In accordance with a further embodiment, the present invention covers compounds of general formula (I), in which Y represents an oxygene atom.
In accordance with a further embodiment, the present invention covers compounds of general formula (I), in which X and Y represent an oxygene atom.
In accordance with a further embodiment, the present invention covers compounds of general formula (I), in which R1 represents halo-C1-C6-alkyl.
In accordance with a further embodiment, the present invention covers compounds of general formula (I), in which R1 represents trifluoromethyl.
In accordance with a further embodiment, the present invention covers compounds of general formula (I), in which R2 represents hydrogen, halogen, cyano or C1-C3-alkyl.
In accordance with a further embodiment, the present invention covers compounds of general formula (I), in which R2 represents hydrogen, fluorine, chlorine, cyano or methyl.
In accordance with a further embodiment, the present invention covers compounds of general formula (I), in which R2 represents hydrogen, chlorine, cyano or C1-C3-alkyl,
In accordance with a further embodiment, the present invention covers compounds of general formula (I), in which R2 represents hydrogen, fluorine, chlorine, methyl or cyano and R3 represents hydrogen.
In accordance with a further embodiment, the present invention covers compounds of general formula (I), in which R3 represents hydrogen.
In accordance with a further embodiment, the present invention covers compounds of general formula (I), in which X and Y represent an oxygene atom, R1 represents trifluoromethyl, R2 represents hydrogen, fluorine, chlorine, methyl or cyano and R3 represents hydrogen.
In accordance with a further embodiment, the present invention covers compounds of general formula (I), in which R4 represents
where “*” denotes the point of attachment to the remainder of the molecule.
In accordance with a further embodiment, the present invention covers compounds of general formula (I), in which R4 represents
where “*” denotes the point of attachment to the remainder of the molecule.
In accordance with a further embodiment, the present invention covers compounds of general formula (I), in which R4 represents hydrogen, C1-C6-alkyl-, which may optionally be mono- or disubstituted by identical or different substituents from the group consisting of halogen, cyano, hydroxy, C1-C3-alkoxy-, —NH2, —NH—(C1-C6-alkyl), —N(C1-C6-alkyl)2, —NH—(C1-C6-alkyl)-C1-C6-alkoxy, C1-C6-alkyl-, C1-C6-alkoxy-, —S(═O)2NH2, —C(═O)—R9, —C(═O)—NH—Rx, —NH—C(═O)—R9, —NH—S(═O)2—R9, —S(═O)2—R9, in which R9 represents methyl, —NH2 or —CH2—CH2—O—CH3 or
where “*” denotes the point of attachment to the remainder of the molecule.
and Rx represents methyl or —CH2—CH2—O—CH3.
In accordance with a further embodiment, the present invention covers compounds of general formula (I), in which R4 represents C1-C6-alkyl-, which may optionally be mono- or disubstituted by identical or different substituents from the group consisting of
where “*” denotes the point of attachment to the remainder of the molecule.
In accordance with a further embodiment, the present invention covers compounds of general formula (I), in which R4 represents C1-C6-alkyl-, which may optionally be mono- or disubstituted by identical or different substituents from the group consisting of
where “*” denotes the point of attachment to the remainder of the molecule.
In accordance with a further embodiment, the present invention covers compounds of general formula (I), in which R4 represents C1-C6-alkyl-, which may optionally be substituted by
where “*” denotes the point of attachment to the remainder of the molecule.
In accordance with a further embodiment, the present invention covers compounds of general formula (I), in which R4 represents C2- or C3-alkyl-, which is substituted by
where “*” denotes the point of attachment to the remainder of the molecule.
In accordance with a further embodiment, the present invention covers compounds of general formula (I), in which R3 and R4 together with the nitrogen atom form the group
where “*” denotes the point of attachment to the remainder of the molecule.
Compounds of most interest are those as follows:
The compounds of general formula (I) of the present invention can be converted to any salt, preferably pharmaceutically acceptable salts, as described herein, by any method which is known to the person skilled in the art. Similarly, any salt of a compound of general formula (I) of the present invention can be converted into the free compound, by any method which is known to the person skilled in the art.
Compounds of general formula (I) of the present invention demonstrate a valuable pharmacological spectrum of action, which could not have been predicted. Compounds of the present invention have surprisingly been found to effectively inhibit MAP4K1 and it is possible therefore that said compounds be used for the treatment or prophylaxis of diseases, preferably cancer or conditions with dysregulated immune responses or other disorders associated with aberrant MAP4K1 signaling, in humans and animals.
Disorders and conditions particularly suitable for treatment with an MAP4K1 inhibitor of the present invention are liquid and solid tumours, such as cancers of the breast, respiratory tract, brain, reproductive organs, digestive tract, urinary tract, eye, liver, skin, head and neck, thyroid, parathyroid and their distant metastases. Those disorders also include lymphomas, sarcomas, and leukaemias.
Examples of breast cancers include, but are not limited to, triple negative breast cancer, invasive ductal carcinoma, invasive lobular carcinoma, ductal carcinoma in situ, and lobular carcinoma in situ.
Examples of cancers of the respiratory tract include, but are not limited to, small-cell and non-small-cell lung carcinoma, as well as bronchial adenoma and pleuropulmonary blastoma.
Examples of brain cancers include, but are not limited to, brain stem and hypophtalmic glioma, cerebellar and cerebral astrocytoma, glioblastoma, medulloblastoma, ependymoma, as well as neuroectodermal and pineal tumour.
Tumours of the male reproductive organs include, but are not limited to, prostate and testicular cancer.
Tumours of the female reproductive organs include, but are not limited to, endometrial, cervical, ovarian, vaginal, and vulvar cancer, as well as sarcoma of the uterus.
Examples of ovarian cancer include, but are not limited to serous tumour, endometrioid tumour, mucinous cystadenocarcinoma, granulosa cell tumour, Sertoli-Leydig cell tumour and arrhenoblastoma.
Examples of cervical cancer include, but are not limited to squamous cell carcinoma, adenocarcinoma, adenosquamous carcinoma, small cell carcinoma, neuroendocrine tumour, glassy cell carcinoma and villoglandular adenocarcinoma.
Tumours of the digestive tract include, but are not limited to, anal, colon, colorectal, esophageal, gallbladder, gastric, pancreatic, rectal, small-intestine, and salivary gland cancers.
Examples of esophageal cancer include, but are not limited to esophageal cell carcinomas and adenocarcinomas, as well as squamous cell carcinomas, leiomyosarcoma, malignant melanoma, rhabdomyosarcoma and lymphoma.
Examples of gastric cancer include, but are not limited to intestinal type and diffuse type gastric adenocarcinoma.
Examples of pancreatic cancer include, but are not limited to ductal adenocarcinoma, adenosquamous carcinomas and pancreatic endocrine tumours.
Tumours of the urinary tract include, but are not limited to, bladder, penile, kidney, renal pelvis, ureter, urethral and human papillary renal cancers.
Examples of kidney cancer include, but are not limited to renal cell carcinoma, urothelial cell carcinoma, juxtaglomerular cell tumour (reninoma), angiomyolipoma, renal oncocytoma, Bellini duct carcinoma, clear-cell sarcoma of the kidney, mesoblastic nephroma and Wilms' tumour. Examples of bladder cancer include, but are not limited to transitional cell carcinoma, squamous cell carcinoma, adenocarcinoma, sarcoma and small cell carcinoma.
Eye cancers include, but are not limited to, intraocular melanoma and retinoblastoma. Examples of liver cancers include, but are not limited to, hepatocellular carcinoma (liver cell carcinomas with or without fibrolamellar variant), cholangiocarcinoma (intrahepatic bile duct carcinoma), and mixed hepatocellular cholangiocarcinoma.
Skin cancers include, but are not limited to, squamous cell carcinoma, Kaposi's sarcoma, malignant melanoma, Merkel cell skin cancer, and non-melanoma skin cancer.
Head-and-neck cancers include, but are not limited to, squamous cell cancer of the head and neck, laryngeal, hypopharyngeal, nasopharyngeal, oropharyngeal cancer, salivary gland cancer, lip and oral cavity cancer and squamous cell.
Lymphomas include, but are not limited to, AIDS-related lymphoma, non-Hodgkin's lymphoma, cutaneous T-cell lymphoma, Burkitt lymphoma, Hodgkin's disease, and lymphoma of the central nervous system.
Sarcomas include, but are not limited to, sarcoma of the soft tissue, osteosarcoma, malignant fibrous histiocytoma, lymphosarcoma, and rhabdomyosarcoma.
Leukemias include, but are not limited to, acute myeloid leukemia, acute lymphoblastic leukemia, chronic lymphocytic leukemia, chronic myelogenous leukemia, and hairy cell leukemia.
The term “treating” or “treatment” as stated throughout this document is used conventionally, for example the management or care of a subject for the purpose of combating, alleviating, reducing, relieving, improving the condition of a disease or disorder, such as a carcinoma. The compounds of the present invention can be used in particular in therapy and prevention, i.e. prophylaxis, of tumour growth and metastases, especially in solid tumours of all indications and stages with or without pre-treatment of the tumour growth.
Generally, the use of chemotherapeutic agents and/or anti-cancer agents in combination with a compound or pharmaceutical composition of the present invention will serve to:
In addition, the compounds of general formula (I) of the present invention can also be used in combination with radiotherapy and/or surgical intervention.
In a further embodiment of the present invention, the compounds of general formula (I) of the present invention may be used to sensitize a cell to radiation, i.e. treatment of a cell with a compound of the present invention prior to radiation treatment of the cell renders the cell more susceptible to DNA damage and cell death than the cell would be in the absence of any treatment with a compound of the present invention. In one aspect, the cell is treated with at least one compound of general formula (I) of the present invention.
Thus, the present invention also provides a method of killing a cell, wherein a cell is administered one or more compounds of the present invention in combination with conventional radiation therapy.
The present invention also provides a method of rendering a cell more susceptible to cell death, wherein the cell is treated with one or more compounds of general formula (I) of the present invention prior to the treatment of the cell to cause or induce cell death. In one aspect, after the cell is treated with one or more compounds of general formula (I) of the present invention, the cell is treated with at least one compound, or at least one method, or a combination thereof, in order to cause DNA damage for the purpose of inhibiting the function of the normal cell or killing the cell.
In other embodiments of the present invention, a cell is killed by treating the cell with at least one DNA damaging agent, i.e. after treating a cell with one or more compounds of general formula (I) of the present invention to sensitize the cell to cell death, the cell is treated with at least one DNA damaging agent to kill the cell. DNA damaging agents useful in the present invention include, but are not limited to, chemotherapeutic agents (e.g. cis platin), ionizing radiation (X-rays, ultraviolet radiation), carcinogenic agents, and mutagenic agents.
In other embodiments, a cell is killed by treating the cell with at least one method to cause or induce DNA damage. Such methods include, but are not limited to, activation of a cell signalling pathway that results in DNA damage when the pathway is activated, inhibiting of a cell signalling pathway that results in DNA damage when the pathway is inhibited, and inducing a biochemical change in a cell, wherein the change results in DNA damage. By way of a non-limiting example, a DNA repair pathway in a cell can be inhibited, thereby preventing the repair of DNA damage and resulting in an abnormal accumulation of DNA damage in a cell.
In one aspect of the invention, a compound of general formula (I) of the present invention is administered to a cell prior to the radiation or other induction of DNA damage in the cell. In another aspect of the invention, a compound of general formula (I) of the present invention is administered to a cell concomitantly with the radiation or other induction of DNA damage in the cell. In yet another aspect of the invention, a compound of general formula (I) of the present invention is administered to a cell immediately after radiation or other induction of DNA damage in the cell has begun.
In another aspect, the cell is in vitro. In another embodiment, the cell is in vivo.
The compounds of the present invention can be administered as the sole pharmaceutical agent or in combination with one or more other pharmaceutically active ingredients where the combination causes no unacceptable adverse effects. The present invention also covers such pharmaceutical combinations. For example, the compounds of the present invention can be combined with: 131I-chTNT, abarelix, abiraterone, aclarubicin, adalimumab, ado-trastuzumab emtansine, afatinib, aflibercept, aldesleukin, alectinib, alemtuzumab, alendronic acid, alitretinoin, altretamine, amifostine, aminoglutethimide, hexyl aminolevulinate, amrubicin, amsacrine, anastrozole, ancestim, anethole dithiolethione, anetumab ravtansine, angiotensin II, antithrombin III, aprepitant, arcitumomab, arglabin, arsenic trioxide, asparaginase, atezolizumab, axitinib, azacitidine, basiliximab, belotecan, bendamustine, besilesomab, belinostat, bevacizumab, bexarotene, bicalutamide, bisantrene, bleomycin, blinatumomab, bortezomib, buserelin, bosutinib, brentuximab vedotin, busulfan, cabazitaxel, cabozantinib, calcitonine, calcium folinate, calcium levofolinate, capecitabine, capromab, carbamazepine carboplatin, carboquone, carfilzomib, carmofur, carmustine, catumaxomab, celecoxib, celmoleukin, ceritinib, cetuximab, chlorambucil, chlormadinone, chlormethine, cidofovir, cinacalcet, cisplatin, cladribine, clodronic acid, clofarabine, cobimetinib, copanlisib, crisantaspase, crizotinib, cyclophosphamide, cyproterone, cytarabine, dacarbazine, dactinomycin, daratumumab, darbepoetin alfa, dabrafenib, dasatinib, daunorubicin, decitabine, degarelix, denileukin diftitox, denosumab, depreotide, deslorelin, dianhydrogalactitol, dexrazoxane, dibrospidium chloride, dianhydrogalactitol, diclofenac, dinutuximab, docetaxel, dolasetron, doxifluridine, doxorubicin, doxorubicin+estrone, dronabinol, eculizumab, edrecolomab, elliptinium acetate, elotuzumab, eltrombopag, endostatin, enocitabine, enzalutamide, epirubicin, epitiostanol, epoetin alfa, epoetin beta, epoetin zeta, eptaplatin, eribulin, erlotinib, esomeprazole, estradiol, estramustine, ethinylestradiol, etoposide, everolimus, exemestane, fadrozole, fentanyl, filgrastim, fluoxymesterone, floxuridine, fludarabine, fluorouracil, flutamide, folinic acid, formestane, fosaprepitant, fotemustine, fulvestrant, gadobutrol, gadoteridol, gadoteric acid meglumine, gadoversetamide, gadoxetic acid, gallium nitrate, ganirelix, gefitinib, gemcitabine, gemtuzumab, Glucarpidase, glutoxim, GM-CSF, goserelin, granisetron, granulocyte colony stimulating factor, histamine dihydrochloride, histrelin, hydroxycarbamide, I-125 seeds, lansoprazole, ibandronic acid, ibritumomab tiuxetan, ibrutinib, idarubicin, ifosfamide, imatinib, imiquimod, improsulfan, indisetron, incadronic acid, ingenol mebutate, interferon alfa, interferon beta, interferon gamma, iobitridol, iobenguane (123I), iomeprol, ipilimumab, irinotecan, Itraconazole, ixabepilone, ixazomib, lanreotide, lansoprazole, lapatinib, lasocholine, lenalidomide, lenvatinib, lenograstim, lentinan, letrozole, leuprorelin, levamisole, levonorgestrel, levothyroxine sodium, lisuride, lobaplatin, lomustine, lonidamine, masoprocol, medroxyprogesterone, megestrol, melarsoprol, melphalan, mepitiostane, mercaptopurine, mesna, methadone, methotrexate, methoxsalen, methylaminolevulinate, methylprednisolone, methyltestosterone, metirosine, mifamurtide, miltefosine, miriplatin, mitobronitol, mitoguazone, mitolactol, mitomycin, mitotane, mitoxantrone, mogamulizumab, molgramostim, mopidamol, morphine hydrochloride, morphine sulfate, nabilone, nabiximols, nafarelin, naloxone+pentazocine, naltrexone, nartograstim, necitumumab, nedaplatin, nelarabine, neridronic acid, netupitant/palonosetron, nivolumab, pentetreotide, nilotinib, nilutamide, nimorazole, nimotuzumab, nimustine, nintedanib, nitracrine, nivolumab, obinutuzumab, octreotide, ofatumumab, olaparib, olaratumab, omacetaxine mepesuccinate, omeprazole, ondansetron, oprelvekin, orgotein, orilotimod, osimertinib, oxaliplatin, oxycodone, oxymetholone, ozogamicine, p53 gene therapy, paclitaxel, palbociclib, palifermin, palladium-103 seed, palonosetron, pamidronic acid, panitumumab, panobinostat, pantoprazole, pazopanib, pegaspargase, PEG-epoetin beta (methoxy PEG-epoetin beta), pembrolizumab, pegfilgrastim, peginterferon alfa-2b, pembrolizumab, pemetrexed, pentazocine, pentostatin, peplomycin, Perflubutane, perfosfamide, Pertuzumab, picibanil, pilocarpine, pirarubicin, pixantrone, plerixafor, plicamycin, poliglusam, polyestradiol phosphate, polyvinylpyrrolidone+sodium hyaluronate, polysaccharide-K, pomalidomide, ponatinib, porfimer sodium, pralatrexate, prednimustine, prednisone, procarbazine, procodazole, propranolol, quinagolide, rabeprazole, racotumomab, radium-223 chloride, radotinib, raloxifene, raltitrexed, ramosetron, ramucirumab, ranimustine, rasburicase, razoxane, refametinib, regorafenib, risedronic acid, rhenium-186 etidronate, rituximab, rolapitant, romidepsin, romiplostim, romurtide, roniciclib, samarium (153Sm) lexidronam, sargramostim, satumomab, secretin, siltuximab, sipuleucel-T, sizofiran, sobuzoxane, sodium glycididazole, sonidegib, sorafenib, stanozolol, streptozocin, sunitinib, talaporfin, talimogene laherparepvec, tamibarotene, tamoxifen, tapentadol, tasonermin, teceleukin, technetium (99mTc) nofetumomab merpentan, 99mTc-HYNIC-[Tyr3]-octreotide, tegafur, tegafur+gimeracil+oteracil, temoporfin, temozolomide, temsirolimus, teniposide, testosterone, tetrofosmin, thalidomide, thiotepa, thymalfasin, thyrotropin alfa, tioguanine, tocilizumab, topotecan, toremifene, tositumomab, trabectedin, trametinib, tramadol, trastuzumab, trastuzumab emtansine, treosulfan, tretinoin, trifluridine+tipiracil, trilostane, triptorelin, trametinib, trofosfamide, thrombopoietin, tryptophan, ubenimex, valatinib, valrubicin, vandetanib, vapreotide, vemurafenib, vinblastine, vincristine, vindesine, vinflunine, vinorelbine, vismodegib, vorinostat, vorozole, yttrium-90 glass microspheres, zinostatin, zinostatin stimalamer, zoledronic acid, zorubicin.
The compounds of the invention can further be combined with other reagents targeting the immune system, such as immune checkpoint inhibitors, e.g. aPD-1/-L1 axis antagonists. PD-1, along with its ligands PD-L1 and PD-L2, function as negative regulators of T cell activation. MAP4K1 suppresses immune cell function. PD-L1 is overexpressed in many cancers and overexpression of PD-1 often occurs concomitantly in tumor infiltrating T cells. Thus results in attenuation of T cell activation and evasion of immune surveillance, which contributes to impaired antitumor immune responses. (Keir M E et al. (2008) Annu. Rev. Immunol. 26:677).
In addition, the inventive compounds can also be used as a therapeutic in a variety of other disorders wherein MAP4K1 is involved such as, cardiovascular and lung diseases. Accordingly, the compounds according to the invention are suitable for the treatment and/or prophylaxis in particular of cardiovascular, inflammatory and fibrotic disorders and of renal disorders, in particular of acute and chronic renal insufficiency, and also of acute and chronic renal failure.
Accordingly, the compounds according to the invention can be used in medicaments for the treatment and/or prophylaxis of cardiovascular, inflammatory and fibrotic disorders, renal disorders, in particular of acute and chronic renal insufficiency, and also of acute and chronic renal failure.
For the purpose of the present invention the term renal insufficiency comprises both acute and chronic manifestations of renal insufficiency, and also underlying or related renal disorders such as diabetic and non-diabetic nephropathies, hypertensive nephropathies, ischaemic renal disorders, renal hypoperfusion, intradialytic hypotension, obstructive uropathy, renal stenoses, glomerulopathies, glomerulonephritis (such as, for example, primary glomerulonephritides; minimal change glomerulonephritis (lipoidnephrosis); membranous glomerulonephritis; focal segmental glomerulosclerosis (FSGS); membrane-proliferative glomerulonephritis; crescentic glomerulonephritis; mesangioproliferative glomerulonephritis (IgA nephritis, Berger's disease); post-infectious glomerulonephritis; secondary glomerulonephritides: diabetes mellitus, lupus erythematosus, amyloidosis, Goodpasture syndrome, Wegener granulomatosis, Henoch-Schönlein purpura, microscopic polyangiitis, acute glomerulonephritis, pyelonephritis (for example as a result of: urolithiasis, benign prostate hyperplasia, diabetes, malformations, abuse of analgesics, Crohn's disease), glomerulosclerosis, arteriolonecrose of the kidney, tubulointerstitial diseases, nephropathic disorders such as primary and congenital or aquired renal disorder, Alport syndrome, nephritis, immunological kidney disorders such as kidney transplant rejection and immunocomplex-induced renal disorders, nephropathy induced by toxic substances, nephropathy induced by contrast agents, diabetic and non-diabetic nephropathy, renal cysts, nephrosclerosis, hypertensive nephrosclerosis and nephrotic syndrome which can be characterized diagnostically, for example by abnormally reduced creatinine and/or water excretion, abnormally elevated blood concentrations of urea, nitrogen, potassium and/or creatinine, altered activity of renal enzymes, for example glutamyl synthetase, altered urine osmolarity or urine volume, elevated microalbuminuria, macroalbuminuria, lesions on glomerulae and arterioles, tubular dilatation, hyperphosphataemia and/or the need for dialysis. The present invention also comprises the use of the compounds according to the invention for the treatment and/or prophylaxis of sequelae of renal insufficiency, for example pulmonary oedema, heart failure, uremia, anemia, electrolyte disturbances (for example hypercalemia, hyponatremia) and disturbances in bone and carbohydrate metabolism.
The present invention also comprises the use of the compounds according to the invention for the treatment and/or prevention of sequelae of renal insufficiency, for example pulmonary oedema, heart failure, uraemia, anaemia, electrolyte disturbances (for example hyperkalaemia, hyponatraemia) and disturbances in bone and carbohydrate metabolism.
The compounds according to the invention are further suitable for the treatment and/or prevention of polycystic kidney disease (PCKD) and of the syndrome of inappropriate ADH secretion (SIADH).
Furthermore, the compounds according to the invention are also suitable for the treatment and/or prophylaxis of metabolic syndrome, hypertension, resistant hypertension, acute and chronic heart failure, coronary heart disease, stable and unstable angina pectoris, peripheral and cardiac vascular disorders, arrhythmias, atrial and ventricular arrhythmias and impaired conduction, for example atrioventricular blocks degrees I-III (AB block I-III), supraventricular tachyarrhythmia, atrial fibrillation, atrial flutter, ventricular fibrillation, ventricular flutter, ventricular tachyarrhythmia, Torsade de pointes tachycardia, atrial and ventricular extrasystoles, AV-junctional extrasystoles, sick sinus syndrome, syncopes, AV-nodal re-entry tachycardia, Wolff-Parkinson-White syndrome, of acute coronary syndrome (ACS), autoimmune cardiac disorders (pericarditis, endocarditis, valvolitis, aortitis, cardiomyopathies), shock such as cardiogenic shock, septic shock and anaphylactic shock, aneurysms, boxer cardiomyopathy (premature ventricular contraction (PVC)), for treatment and/or prophylaxis of thromboembolic disorders and ischaemias such as myocardial ischaemia, myocardial infarction, stroke, cardiac hypertrophy, transient and ischaemic attacks, preeclampsia, inflammatory cardiovascular disorders, spasms of the coronary arteries and peripheral arteries, oedema formation, for example pulmonary oedema, cerebral oedema, renal oedema or oedema caused by heart failure, peripheral circulatory disturbances, reperfusion damage, arterial and venous thromboses, myocardial insufficiency, endothelial dysfunction, to prevent restenoses, for example after thrombolysis therapies, percutaneous transluminal angioplasties (PTA), transluminal coronary angioplasties (PTCA), heart transplants and bypass operations, and also micro- and macrovascular damage (vasculitis), increased levels of fibrinogen and of low-density lipoprotein (LDL) and increased concentrations of plasminogen activator inhibitor 1 (PAI-1), and also for treatment and/or prophylaxis of erectile dysfunction and female sexual dysfunction.
In addition, the compounds according to the invention are also suitable for treatment and/or prophylaxis of asthmatic disorders, pulmonary arterial hypertension (PAH) and other forms of pulmonary hypertension (PH) including left-heart disease, HIV, sickle cell anaemia, thromboembolisms (CTEPH), sarcoidosis, COPD or pulmonary fibrosis-associated pulmonary hypertension, chronic-obstructive pulmonary disease (COPD), acute respiratory distress syndrome (ARDS), acute lung injury (ALI), alpha-1-antitrypsin deficiency (AATD), pulmonary fibrosis, pulmonary emphysema (for example pulmonary emphysema induced by cigarette smoke) and cystic fibrosis (CF).
The compounds described in the present invention are also active compounds for control of central nervous system disorders characterized by disturbances of the NO/cGMP system. They are suitable in particular for improving perception, concentration, learning or memory after cognitive impairments like those occurring in particular in association with situations/diseases/syndromes such as mild cognitive impairment, age-associated learning and memory impairments, age-associated memory losses, vascular dementia, craniocerebral trauma, stroke, dementia occurring after strokes (post stroke dementia), post-traumatic craniocerebral trauma, general concentration impairments, concentration impairments in children with learning and memory problems, Alzheimer's disease, Lewy body dementia, dementia with degeneration of the frontal lobes including Pick's syndrome, Parkinson's disease, progressive dementia with corticobasal degeneration, amyolateral sclerosis (ALS), Huntington's disease, demyelinization, multiple sclerosis, thalamic degeneration, Creutzfeld-Jacob dementia, HIV dementia, schizophrenia with dementia or Korsakoff's psychosis. They are also suitable for treatment and/or prophylaxis of central nervous system disorders such as states of anxiety, tension and depression, CNS-related sexual dysfunctions and sleep disturbances, and for controlling pathological disturbances of the intake of food, stimulants and addictive substances.
The compounds according to the invention are furthermore also suitable for controlling cerebral blood flow and thus represent effective agents for controlling migraines. They are also suitable for the prophylaxis and control of sequelae of cerebral infarction (cerebral apoplexy) such as stroke, cerebral ischaemia and craniocerebral trauma. The compounds according to the invention can likewise be used for controlling states of pain and tinnitus. In addition, the compounds according to the invention have anti-inflammatory action and can therefore be used as anti-inflammatory agents for treatment and/or prophylaxis of sepsis (SIRS), multiple organ failure (MODS, MOF), inflammatory disorders of the kidney, chronic intestinal inflammations (IBD, Crohn's disease, UC), pancreatitis, peritonitis, rheumatoid disorders, inflammatory skin disorders and inflammatory eye disorders.
Furthermore, the compounds according to the invention can also be used for treatment and/or prophylaxis of autoimmune diseases.
The compounds according to the invention are also suitable for treatment and/or prophylaxis of fibrotic disorders of the internal organs, for example the lung, the heart, the kidney, the bone marrow and in particular the liver, and also dermatological fibroses and fibrotic eye disorders. In the context of the present invention, the term fibrotic disorders includes in particular the following terms: hepatic fibrosis, cirrhosis of the liver, pulmonary fibrosis, endomyocardial fibrosis, nephropathy, glomerulonephritis, interstitial renal fibrosis, fibrotic damage resulting from diabetes, bone marrow fibrosis and similar fibrotic disorders, scleroderma, morphea, keloids, hypertrophic scarring (also following surgical procedures), naevi, diabetic retinopathy, proliferative vitroretinopathy and disorders of the connective tissue (for example sarcoidosis). The compounds according to the invention are also suitable for controlling postoperative scarring, for example as a result of glaucoma operations.
The compounds according to the invention can also be used cosmetically for ageing and keratinized skin.
Moreover, the compounds according to the invention are suitable for treatment and/or prophylaxis of hepatitis, neoplasms, osteoporosis, glaucoma and gastroparesis.
The present invention further provides the use of the compounds according to the invention for treatment and/or prophylaxis of disorders, especially the disorders mentioned above.
The present invention further provides the use of the compounds according to the invention for the treatment and/or prophylaxis of chronic renal disorders, acute and chronic renal insufficiency, diabetic, inflammatory or hypertensive nephropathies, fibrotic disorders, cardiac insufficiency, angina pectoris, hypertension, pulmonary hypertension, ischemias, vascular disorders, thromboembolic disorders, arteriosclerosis, sickle cell anemia, erectile dysfunction, benign prostate hyperplasia, dysuria associated with benign prostate hyperplasia, Huntington, dementia, Alzheimer and Creutzfeld-Jakob.
The present invention further provides a method for treatment and/or prophylaxis of disorders, in particular the disorders mentioned above, using an effective amount of at least one of the compounds according to the invention.
The present invention further provides a method for the treatment and/or prophylaxis of chronic renal disorders, acute and chronic renal insufficiency, diabetic, inflammatory or hypertensive nephropathies, fibrotic disorders, cardiac insufficiency, angina pectoris, hypertension, pulmonary hypertension, ischemias, vascular disorders, thromboembolic disorders, arteriosclerosis, sickle cell anemia, erectile dysfunction, benign prostate hyperplasia, dysuria associated with benign prostate hyperplasia, Huntington, dementia, Alzheimer and Creutzfeld-Jakob.
In another embodiment, the inventive compounds can also be used to treat or to prevent uterine fibroids (uterine leiomyoma or uterine myoma) in women.
Uterine fibroids are benign tumors of the myometrium, the smooth muscle layer of the uterus. Uterine fibroids grow slowly during a women's life, and their growth is dependent on the female sexual hormones estradiol and progesterone [Kawaguchi K et al. Immunohistochemical analysis of oestrogen receptors, progesterone receptors and Ki-67 in leiomyoma and myometrium during the menstrual cycle and pregnancy Virchows Arch A Pathol Anat Histopathol. 1991; 419(4):309-15.], therefore the highest prevalence of uterine fibroids with approx. 70% and >80% in white and afro-american women, respectively, is found from 35 years of age onwards to menopause, when they shrink due to reduced hormone levels [Baird D D et al. High cumulative incidence of uterine leiomyoma in black and white women: Ultrasound evidence Am J Obstet Gynecol. 2003 January; 188(1):100-7.]. Approx 30% and 45% of white and afro-american women, respectively, do show clinically relevant symptoms due to their fibroids, which are heavy menstrual bleeding and pain, which is related to the menstrual cycle [David M et al. Myoma-associated pain frequency and intensity: a retrospective evaluation of 1548 myoma patients. Eur J Obstet Gynecol Reprod Biol. 2016 April; 199:137-40]. Heavy menstrual bleeding in this respect is defined by a blood loss of more than 80 mL in a menstrual bleeding period [Fraser I S et al. The FIGO Recommendations on Terminologies and Definitions for Normal and Abnormal Uterine Bleeding, Semin Reprod Med 2011; 29(5): 383-390]. Submucosal position of the uterine fibroids, e.g. those located directly below the endometrium, seems to have an even more severe effect on uterine bleeding, which may result in anemia in affected women [Yang J H et al. Impact of submucous myoma on the severity of anemia. Fertil Steril. 2011 April; 95(5):1769-72]. Furthermore, uterine fibroids, due to their symptoms, do severly affect the quality of life of affected women [Downes E et al. The burden of uterine fibroids in five European countries. Eur J Obstet Gynecol Reprod Biol. 2010 September; 152(1):96-102].
Compounds of the present invention can be utilized to inhibit, block, reduce or decrease MAP4K1 activation by exogenous and/or endogenous ligands for the reduction of tumour growth and the modulation of dysregulated immune responses e.g. to block immunosuppression and increase immune cell activation and infiltration in the context of cancer and cancer immunotherapy; This method comprises administering to a mammal in need thereof, including a human, an amount of a compound of this invention, or a pharmaceutically acceptable salt, isomer, polymorph, metabolite, hydrate, solvate or ester thereof; which is effective to treat the disorder.
The present invention also provides methods of treating a variety of other disorders wherein MAP4K1 is involved such as, but not limited to, disorders with dysregulated immune responses, inflammation, vaccination for infection & cancer, viral infections, obesity and diet-induced obesity, adiposity, metabolic disorders, hepatic steatosis and uterine fibroids. These disorders have been well characterized in humans, but also exist with a similar etiology in other mammals, and can be treated by administering pharmaceutical compositions of the present invention.
In accordance with a further aspect, the present invention covers compounds of general formula (I), as described supra, or stereoisomers, tautomers, N-oxides, hydrates, solvates, and salts thereof, particularly pharmaceutically acceptable salts thereof, or mixtures of same, for use in the treatment or prophylaxis of diseases, in particular cancer or conditions with dysregulated immune responses or other disorders associated with aberrant MAP4K1 signaling.
The pharmaceutical activity of the compounds according to the invention can be explained by their activity as MAP4K1 inhibitors.
In accordance with a further aspect, the present invention covers the use of compounds of general formula (I), as described supra, or stereoisomers, tautomers, N-oxides, hydrates, solvates, and salts thereof, particularly pharmaceutically acceptable salts thereof, or mixtures of same, for the treatment or prophylaxis of diseases, in particular cancer or conditions with dysregulated immune responses or other disorders associated with aberrant MAP4K1 signaling, particularly liquid and solid tumours.
In accordance with a further aspect, the present invention covers the compounds of general formula (I), as described supra, or stereoisomers, tautomers, N-oxides, hydrates, solvates, and salts thereof, particularly pharmaceutically acceptable salts thereof, or mixtures of same, for the use of treatment or prophylaxis of diseases, in particular cancer or conditions with dysregulated immune responses or other disorders associated with aberrant MAP4K1 signaling, particularly liquid and solid tumours.
In accordance with a further aspect, the present invention covers the use of compounds of general formula (I), as described supra, or stereoisomers, tautomers, N-oxides, hydrates, solvates, and salts thereof, particularly pharmaceutically acceptable salts thereof, or mixtures of same, in a method of treatment or prophylaxis of diseases, in particular cancer or conditions with dysregulated immune responses or other disorders associated with aberrant MAP4K1 signaling, particularly liquid and solid tumours.
In accordance with a further aspect, the present invention covers use of a compound of general formula (I), as described supra, or stereoisomers, tautomers, N-oxides, hydrates, solvates, and salts thereof, particularly pharmaceutically acceptable salts thereof, or mixtures of same, for the preparation of a pharmaceutical composition, preferably a medicament, for the prophylaxis or treatment of diseases, in particular cancer or conditions with dysregulated immune responses or other disorders associated with aberrant MAP4K1 signaling, particularly liquid and solid tumours.
In accordance with a further aspect, the present invention covers a method of treatment or prophylaxis of diseases, in particular cancer or conditions with dysregulated immune responses or other disorders associated with aberrant MAP4K1 signaling, particularly liquid and solid tumours, using an effective amount of a compound of general formula (I), as described supra, or stereoisomers, tautomers, N-oxides, hydrates, solvates, and salts thereof, particularly pharmaceutically acceptable salts thereof, or mixtures of same.
In accordance with a further aspect, the present invention covers pharmaceutical compositions, in particular a medicament, comprising a compound of general formula (I), as described supra, or a stereoisomer, a tautomer, an N-oxide, a hydrate, a solvate, a salt thereof, particularly a pharmaceutically acceptable salt, or a mixture of same, and one or more excipients), in particular one or more pharmaceutically acceptable excipient(s). Conventional procedures for preparing such pharmaceutical compositions in appropriate dosage forms can be utilized.
The present invention furthermore covers pharmaceutical compositions, in particular medicaments, which comprise at least one compound according to the invention, conventionally together with one or more pharmaceutically suitable excipients, and to their use for the above mentioned purposes.
It is possible for the compounds according to the invention to have systemic and/or local activity. For this purpose, they can be administered in a suitable manner, such as, for example, via the oral, parenteral, pulmonary, nasal, sublingual, lingual, buccal, rectal, vaginal, dermal, transdermal, conjunctival, otic route or as an implant or stent.
For these administration routes, it is possible for the compounds according to the invention to be administered in suitable administration forms.
For oral administration, it is possible to formulate the compounds according to the invention to dosage forms known in the art that deliver the compounds of the invention rapidly and/or in a modified manner, such as, for example, tablets (uncoated or coated tablets, for example with enteric or controlled release coatings that dissolve with a delay or are insoluble), orally-disintegrating tablets, films/wafers, films/lyophylisates, capsules (for example hard or soft gelatine capsules), sugar-coated tablets, granules, pellets, powders, emulsions, suspensions, aerosols or solutions. It is possible to incorporate the compounds according to the invention in crystalline and/or amorphised and/or dissolved form into said dosage forms.
Parenteral administration can be effected with avoidance of an absorption step (for example intravenous, intraarterial, intracardial, intraspinal or intralumbal) or with inclusion of absorption (for example intramuscular, subcutaneous, intracutaneous, percutaneous or intraperitoneal). Administration forms which are suitable for parenteral administration are, inter alia, preparations for injection and infusion in the form of solutions, suspensions, emulsions, lyophylisates or sterile powders.
Examples which are suitable for other administration routes are pharmaceutical forms for inhalation [inter alia powder inhalers, nebulizers], nasal drops, nasal solutions, nasal sprays; tablets/films/wafers/capsules for lingual, sublingual or buccal administration; suppositories; eye drops, eye ointments, eye baths, ocular inserts, ear drops, ear sprays, ear powders, ear-rinses, ear tampons; vaginal capsules, aqueous suspensions (lotions, mixturae agitandae), lipophilic suspensions, emulsions, ointments, creams, transdermal therapeutic systems (such as, for example, patches), milk, pastes, foams, dusting powders, implants or stents.
The compounds according to the invention can be incorporated into the stated administration forms. This can be effected in a manner known per se by mixing with pharmaceutically suitable excipients. Pharmaceutically suitable excipients include, inter alia,
The present invention furthermore relates to a pharmaceutical composition which comprise at least one compound according to the invention, conventionally together with one or more pharmaceutically suitable excipient(s), and to their use according to the present invention. In accordance with another aspect, the present invention covers pharmaceutical combinations, in particular medicaments, comprising at least one compound of general formula (I) of the present invention and at least one or more further active ingredients, in particular for the treatment and/or prophylaxis of cancer or conditions with dysregulated immune responses or other disorders associated with aberrant MAP4K1 signaling generic name disorders, particularly liquid and solid tumours.
The term “combination” in the present invention is used as known to persons skilled in the art, it being possible for said combination to be a fixed combination, a non-fixed combination or a kit-of-parts.
A “fixed combination” in the present invention is used as known to persons skilled in the art and is defined as a combination wherein, for example, a first active ingredient, such as one or more compounds of general formula (I) of the present invention, and a further active ingredient are present together in one unit dosage or in one single entity. One example of a “fixed combination” is a pharmaceutical composition wherein a first active ingredient and a further active ingredient are present in admixture for simultaneous administration, such as in a formulation. Another example of a “fixed combination” is a pharmaceutical combination wherein a first active ingredient and a further active ingredient are present in one unit without being in admixture.
A non-fixed combination or “kit-of-parts” in the present invention is used as known to persons skilled in the art and is defined as a combination wherein a first active ingredient and a further active ingredient are present in more than one unit. One example of a non-fixed combination or kit-of-parts is a combination wherein the first active ingredient and the further active ingredient are present separately. It is possible for the components of the non-fixed combination or kit-of-parts to be administered separately, sequentially, simultaneously, concurrently or chronologically staggered.
Based upon standard laboratory techniques known to evaluate compounds useful for the treatment of cancer or conditions with dysregulated immune responses or other disorders associated with aberrant MAP4K1 signaling, by standard toxicity tests and by standard pharmacological assays for the determination of treatment of the conditions identified above in mammals, and by comparison of these results with the results of known active ingredients or medicaments that are used to treat these conditions, the effective dosage of the compounds of the present invention can readily be determined for treatment of each desired indication. The amount of the active ingredient to be administered in the treatment of one of these conditions can vary widely according to such considerations as the particular compound and dosage unit employed, the mode of administration, the period of treatment, the age and sex of the patient treated, and the nature and extent of the condition treated.
The total amount of the active ingredient to be administered will generally range from about 0.001 mg/kg to about 200 mg/kg body weight per day, and preferably from about 0.01 mg/kg to about 20 mg/kg body weight per day. Clinically useful dosing schedules will range from one to three times a day dosing to once every four weeks dosing. In addition, it is possible for “drug holidays”, in which a patient is not dosed with a drug for a certain period of time, to be beneficial to the overall balance between pharmacological effect and tolerability. It is possible for a unit dosage to contain from about 0.5 mg to about 1500 mg of active ingredient, and can be administered one or more times per day or less than once a day. The average daily dosage for administration by injection, including intravenous, intramuscular, subcutaneous and parenteral injections, and use of infusion techniques will preferably be from 0.01 to 200 mg/kg of total body weight. The average daily rectal dosage regimen will preferably be from 0.01 to 200 mg/kg of total body weight. The average daily vaginal dosage regimen will preferably be from 0.01 to 200 mg/kg of total body weight. The average daily topical dosage regimen will preferably be from 0.1 to 200 mg administered between one to four times daily. The transdermal concentration will preferably be that required to maintain a daily dose of from 0.01 to 200 mg/kg. The average daily inhalation dosage regimen will preferably be from 0.01 to 100 mg/kg of total body weight.
Of course the specific initial and continuing dosage regimen for each patient will vary according to the nature and severity of the condition as determined by the attending diagnostician, the activity of the specific compound employed, the age and general condition of the patient, time of administration, route of administration, rate of excretion of the drug, drug combinations, and the like. The desired mode of treatment and number of doses of a compound of the present invention or a pharmaceutically acceptable salt or ester or composition thereof can be ascertained by those skilled in the art using conventional treatment tests.
NMR peak forms are stated as they appear in the spectra, possible higher order effects have not been considered. The multiplicities are stated according to the signal form which appears in the spectrum, NMR-spectroscopic effects of a higher order were not taken into consideration. Multiplicity of the NMR signals: s=singlet, d=doublet, t=triplet, q=quartet, quin=quintet, br=broad signal, m=multiplet. NMR signals: shift in [ppm]. Combinations of multiplicity could be e.g. dd=doublet from doublet.
Chemical names were generated using the ACD/Name software from ACD/Labs. In some cases generally accepted names of commercially available reagents were used in place of ACD/Name generated names.
Table 1 lists the abbreviations used in this paragraph and in the Examples section as far as they are not explained within the text body. Other abbreviations have their meanings customary per se to the skilled person.
The various aspects of the invention described in this application are illustrated by the following examples which are not meant to limit the invention in any way.
The example testing experiments described herein serve to illustrate the present invention and the invention is not limited to the examples given.
All reagents, for which the synthesis is not described in the experimental part, are either commercially available, or are known compounds or may be formed from known compounds by known methods by a person skilled in the art.
The compounds and intermediates produced according to the methods of the invention may require purification. Purification of organic compounds is well known to the person skilled in the art and there may be several ways of purifying the same compound. In some cases, no purification may be necessary. In some cases, the compounds may be purified by crystallization. In some cases, impurities may be stirred out using a suitable solvent. In some cases, the compounds may be purified by chromatography, particularly flash column chromatography, using for example prepacked silica gel cartridges, e.g. Biotage SNAP cartidges KP-Sil® or KP-NH® in combination with a Biotage autopurifier system (SP4® or Isolera Four®) and eluents such as gradients of hexane/ethyl acetate or DCM/methanol. In some cases, the compounds may be purified by preparative HPLC using for example a Waters autopurifier equipped with a diode array detector and/or on-line electrospray ionization mass spectrometer in combination with a suitable prepacked reverse phase column and eluents such as gradients of water and acetonitrile which may contain additives such as trifluoroacetic acid, formic acid or aqueous ammonia.
In some cases, purification methods as described above can provide those compounds of the present invention which possess a sufficiently basic or acidic functionality in the form of a salt, such as, in the case of a compound of the present invention which is sufficiently basic, a trifluoroacetate or formate salt for example, or, in the case of a compound of the present invention which is sufficiently acidic, an ammonium salt for example. A salt of this type can either be transformed into its free base or free acid form, respectively, by various methods known to the person skilled in the art, or be used as salts in subsequent biological assays. It is to be understood that the specific form (e.g. salt, free base etc.) of a compound of the present invention as isolated and as described herein is not necessarily the only form in which said compound can be applied to a biological assay in order to quantify the specific biological activity.
The following paragraphs outline a variety of synthetic approaches suitable to prepare compounds of the general formula (I), and intermediates useful for their synthesis. In addition to the routes described below, also other routes may be used to synthesise the target compounds, in accordance with common general knowledge of a person skilled in the art of organic synthesis. The order of transformations exemplified in the following schemes is therefore not intended to be limiting, and suitable synthesis steps from various schemes can be combined to form additional synthesis sequences. In addition, interconversion of any of the substituents, in particular R1, R2, R3 and R4, which are as defined in formula (I) supra, can be achieved before and/or after the exemplified transformations. These modifications can be, for example, the introduction of protective groups, cleavage of protective groups, reduction or oxidation of functional groups, halogenation, metallation, metal catalysed coupling reactions, exemplified by but not limited to e.g. Buchwald, Suzuki, Sonogashira and Ullmann coupling, ester saponifications, amide coupling reactions, and/or substitution or other reactions known to a person skilled in the art. These transformations include those which introduce a functionality allowing for further interconversion of substituents. Appropriate protective groups and their introduction and cleavage are well-known to a person skilled in the art (see for example T. W. Greene and P. G. M. Wuts in Protective Groups in Organic Synthesis, 4th edition, Wiley 2006). Further, it is possible that two or more successive steps may be performed without work-up being performed between said steps, e.g. a “one-pot” reaction, as it is well-known to a person skilled in the art.
Compounds of general formula (I) can be assembled according to Scheme 1, by reaction of amine derivatives of formula (II), in which R1, R2 and X are as defined for the compounds of general formula (I), and a second amine derivative (Ill), in which R3, and R4 are as defined for the compounds of general formula (I), by means of urea formation well known to the person skilled in the art. Said urea formation can be performed by reaction of compounds of the formula (II) with the intermediacy of a formed and possibly isolated isocyanate or isothiocyantate (IVa) using a suitable reagent such as 1,1′-carbonylbis-1H-imidazole, di- or triphosgene for Y═O, or as thiophosgene or 1,1′-thiocarbonylbis-1H-imidazole for Y═S. Compounds of general formula (I) can also be assembled by conversion of amine derivatives of formula (II) to an intermediately formed and possibly isolated carbamate or thiocarbamate (IVb) using a suitable reagent such as phenyl chloroformate or O-phenyl chlorothionoformate in which Z is H, NO2, or perfluoro in an appropriate solvent such as tetrahydrofuran, dichloromethane, or ethylacetate in the presence of an appropriate base such as pyridine, sodium hydrogencarbonate, or triethylamine. This intermediate IV(b) is then reacted with the second amine derivative (Ill) in an appropriate solvent such as pyridine, or dimethylformamide.
In a similar way the compounds of general formula (I) can be assembled using the amine (111) as starting material. With the former described reaction the amine (III) can react to the intermediately formed isocyanate or isothiocyanate (Va for R3═H), if it is not commercially available, in which R4 and Y are as defined for the compounds of general formula (I) or the carbamate or thiocarbamate (Vb) using a suitable reagent such as phenyl chloroformate or O-phenyl chlorothionoformate in which Z is H, NO2, or perfluoro and R3, R4 and Y are as defined for the compounds of general formula (I) with the second amine (11).
Depending on the choice of protecting group PG in formula (II), (IVa), and (IVb), which is preferentially trimethylsilylethyloxymethyl (SEM), but can be any other protecting group well known to the person skilled in the art, the deprotection can be performed using trifluoroacetic acid in the case of trimethylsilylethyloxymethyl, in an inert solvent such as dichloromethane, within a temperature range from 0° C. to the boiling point of the used solvent. The deprotection in the case of trimethylsilylethyloxymethyl can be also performed using tetra-butylammonium fluoride in the presence of ethylenediamine in an inert solvent such as tetrahydrofuran within a temperature range from 0° C. to the boiling point of the used solvent.
Preferred herein is the performance of said urea formation using the amine (II) and the intermediately formed and maybe isolated carbamate (IVb) and the subsequent reaction with the respective second amine (III) in DMF or using the amine (II) in a reaction with the respective cyanate or isothiocyanate (Va) in pyridine as a solvent, within a temperature range from 0° C. to 100° C.
The amine intermediates of formula (II) are known to the person skilled in the art and can, if not commercially available, be prepared according to Schemes 2 and 3 shown below. The second amine derivatives of formula (III) are either commercially available in some structural variety, or they can be prepared using synthetic methods described in many textbooks such as March's Advanced Organic Chemistry: Reactions, Mechanisms, and Structure, 7th Edition.
As illustrated in Scheme 2, the amine derivatives of formula (II) are known, commercially available, or can be prepared from the commercially available heterocyle of the formula (VI), in which R1a represents a hydrogen or a trifluoromethyl group. Said heterocycle of the formula (VI) can be reacted with nitric acid, in the case of leaving group (LG)=NO2, or in a two step sequence for LG=C; using MCPBA to form a 7-N-oxide, which then reacts in a subsequent step with methanesulfonyl chloride to give intermediate heterocycles of formula (VII).
Compounds of the formula (VII) are converted to chlorides of formula (VIII) using a two step sequence, which begins with production of a 7-N-oxide which then reacts in a subsequent step with trichloroacetyl chloride in the presence of HMDS. Said heterocycles of formula (VIII) can be transformed to the protected intermediates of the formula (IX) using an apprioate reagent such as trimetylsilylethoxymethyl chloride, triisopropylsilyl chloride or trityl chloride or other reagents known to a person skilled in the art. Preferably, trimetylsilylethoxymethyl chloride is used, in the presence of a base such as sodium hydride, triethyl amine, or ethyl diisopropyl amine in an inert solvent such as THF, DMSO or DMF.
Protected heterocycles of the formula (IX) are then reacted with a compound of the formula (X) in the presence of sodium hydride or an alkali carbonate, such as sodium carbonate, potassium carbonate, or cesium carbonate, in a suitable solvent such as DMSO or DMF, as well known to the person skilled in the art, to give compounds of formula (XI). For compounds of formula (X) with X=NH it may be necessary to protect the other amino function in an intermediate fashion.
Dechlorination is preferentially performed using a hydrogen atmosphere and palladium on carbon as catalyst in an inert solvent such as ethanol, ethyl acetate or dichloromethane at 20-50° C. as described in Org. Process Res. Dev. 2010, page 168-173, to give a subset of the amines of the formula (II) named (IIa) in which R1a are as defined for compounds of general formula (VI).
All other amines of the subset (IIb) from the general formula (II) can be assembled from the amine (IIa) with R1a=H according to Scheme 3.
Protection of the free amine in (IIa) with R1a=H using trifluoroacetic anhydride in an inert solvent such as dichloromethane in the presence of an tertiary amine such as triethyl amine or ethyl diisopropyl amine, yields the amides of the general formula (XII) in which R2 is as defined for the compounds of general formula (I). Said amides of formula (XII) can be converted into halogen substituted compounds of the general formula (XIII) with R1b=Cl, Br, or I using the corresponding N-halo-succinimide in an inert solvent such as dichloromethane or tetrachloromethane. In the case of compounds of the general formula (XIII) with R1b=CN, and C1-C6-alkyl, a subsequent reaction of a said halogen compound of the general formula (XII) with R1b=Br or I is used as starting material. For formation of the corresponding nitrile, a reaction using cuprous cyanide in an inert solvent such as dimethyl formamide or dimethyl acetamide at elevated temperatures, for example between 90-120° C. is required. Furthermore, a palladium-catalyzed method known to the person skilled in the art, for example, with zinc cyanide or potassium ferrocyanide as described in Chem. Soc. Rev., 2011, 40, 5049-5067 can be utilized. For the preparation of compounds of the general formula (XIII) with R1b=C1-C6-alkyl a Suzuki reaction known to the person skilled in the art is used as described, for example in Metal-Catalyzed Cross-Coupling Reactions, Second Edition (Editors: Armin de Meijere, Francois Diederich, Wiley-VCH) or in Catal. Lett. (2016) page 820-840. After saponification of compounds of the general formula (XIII) using lithium, sodium or potassium hydroxide, the compounds of the general formula (IIb) are obtained. Taken together, formulae (IIa) and (IIb) constitute formula (II).
A complementary approach to compounds of formula (II) is described in Scheme 4.
The respective fluoro-nitrobenzene derivatives of formula (XIII) are reacted with 4-hydroxy-7-azaindole (XII) (e.g. US2007/238726), in a suitable solvent system, for example dimethylsulfoxide, in the presence of a base, for example potassium carbonate or cesium carbonate, in a temperature range from room temperature to the boiling point of the respective solvent. Preferable the reaction is carried out at room temperature to furnish intermediates of general formula (XIV).
Intermediates of general formula (XIV), are then protected with an appropriate protecting group, for example using an appropriate reagent such as trimetylsilylethoxymethyl chloride, triisopropylsilyl chloride or trityl chloride or other reagents known to a person skilled in the art. Preferably, trimetylsilylethoxymethyl chloride is used, in the presence of a base such as sodium hydride, triethylamine, or ethyldiisopropylamine in an inert solvent such as tetrahydrofuran, dimethylsulfoxide or dimethylformamide to afford compounds of formula (XV).
Compounds of formula (XV) can be converted into halogen substituted compounds of the general formula (XVI) with R1=Cl, Br, or I using the corresponding N-halo-succinimide in an inert solvent such as dichloromethane, dimethylformamide, or tetrachloromethane, in a temperature range from room temperature to the boiling point of the respective solvent. Preferable the reaction is carried out at room temperature to furnish intermediates of general formula (XVI).
Compounds of general formula (XVI), where R1=I, can be reacted with trifluoromethylation reagents, for example diphenyl(trifluoromethyl)sulfonium trifluoromethanesulfonate as described in Angew. Chem. Int. Ed. 2011, 50, pg. 1896-1900, or any other reagent know to one skilled in the art, to afford compounds of general formula (XVI) where R1=CF3. Preferably, diphenyl(trifluoromethyl)sulfonium trifluoromethanesulfonate is used, in the presence of additives, such as copper(0), in a solvent such as dimethylformamide, in a temperature range from room temperature to the boiling point of the respective solvent. Ideally, the reaction is carried out at 60° C. to furnish intermediates of general formula (XVI) where R1=CF3.
In the case of compounds of the general formula (XVI) with R1=CN, and C1-C6-alkyl, a subsequent reaction of a said halogen compounds of the general formula (XVI) with R1=Br or I is required. For formation of the corresponding nitrile, a reaction using cuprous cyanide in an inert solvent such as dimethyl formamide or dimethyl acetamide at elevated temperatures, for example between 90-120° C. is required. Furthermore, a palladium-catalyzed method known to the person skilled in the art, for example, with zinc cyanide or potassium ferrocyanide as described in Chem. Soc. Rev., 2011, 40, 5049-5067 can be utilized. For the preparation of compounds of the general formula (XVI) with R1=C1-C6-alkyl a Suzuki reaction known to the person skilled in the art is used as described, for example in Metal-Catalyzed Cross-Coupling Reactions, Second Edition (Editors: Armin de Meijere, Francois Diederich, Wiley-VCH) or in Catal. Lett. (2016) page 820-840.
Reduction of the nitro functionality contained within compounds of formula (XVI) affords amines of formula (II). Preferentially, the reaction is performed with the addition of a reducing agent, for example Iron(0), with additives such as ammonium chloride, in an appropriate mixture of solvents, for example a mixture of water, tetrahydrofuran, and methanol, in a temperature range from room temperature to the boiling point of the respective solvent. Ideally the reaction is performed at 80° C. to furnish amine intermediates of general formula (II). The reaction can be carried out using alternative reducing agents to those skilled in the art, for example tin(II)chloride, in an appropriate solvent, such as methanol, in a temperature range from room temperature to the boiling point of the respective solvent. Ideally the reaction is performed at room temperature to furnish amine intermediates of general formula (II).
Method 1:
(prep. HPLC) System: Labomatic, Pump: HD-5000, Fraction Collector: LABOCOL Vario-4000, UV-Detector: Knauer UVD 2.1S; Column: Chromatorex RP C18 10 μm 125×30 mm; Solvent: A=water+0.1% Vol. ammonia (99%), B=Acetonitril; Flow: 150 mL/min; temperature: room temperature
A solution of 6-chloro-4-nitro-3-(trifluoromethyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}-1H-pyrrolo[2,3-b]pyridine (500 mg, 1.26 mmol, see Synthesis 2007, page 251-258, Org. Process Res. Dev. 2010, page 168-173), 4-aminophenol (207 mg, 1.89 mmol) and potassium carbonate (524 mg, 3.79 mmol) in DMSO (5.0 mL) was stirred at 120° C. for 3 hours. After cooling to room temperature the reaction mixture was diluted with ethyl actate (200 mL). This organic phase was washed two times with water (30 mL) and once with brine (20 mL), then dried over sodium sulfate and after filtration dried to dryness. The resulting residue was purified via a Biotage chromatography system (28 g snap KP-NH column, hexane/0-70% ethyl acetate) to obtain 479 mg (100% purity, 83% yield) of the desired title compound.
1H-NMR (400 MHz, DMSO-d6) δ [ppm]: −0.10-−0.06 (m, 9H), 0.81-0.89 (m, 2H), 3.53-3.60 (m, 2H), 5.24 (s, 2H), 5.60 (s, 2H), 6.31 (s, 1H), 6.63-6.68 (m, 2H), 6.89-6.94 (m, 2H), 8.32 (d, 1H).
To a solution of 4-{[6-chloro-3-(trifluoromethyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}-1H-pyrrolo[2,3-b]pyridin-4-yl]oxy}aniline (474 mg, 1.04 mmol, intermediate 1) and triethylamine (170 μL, 1.2 mmol) in ethanol (35 mL) was given 10% Pd on carbon (47.4 mg). This mixture was stirred in an hydrogen atmosphere for 7 hours at room temperature. Then the mixture was filtered through Celite and the Celite was washed with ethyl acetate. The organic phase was evaporated to dryness and the resulting residue was purified via a Biotage chromatography system (28 g snap KP-NH column, hexane/0-70% ethyl acetate) to obtain 424 mg (100% purity, 97% yield) of the desired title compound.
1H-NMR (400 MHz, DMSO-d6) δ [ppm]: −0.10-−0.07 (m, 9H), 0.79-0.87 (m, 2H), 3.53-3.59 (m, 2H), 5.16 (s, 2H), 5.66 (s, 2H), 6.41 (d, 1H), 6.62-6.67 (m, 2H), 6.85-6.90 (m, 2H), 8.20-8.22 (m, 1H), 8.21 (d, 1H), 8.26 (s, 1H).
To solution of using 4-{[3-(trifluoromethyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}-1H-pyrrolo[2,3-b]pyridin-4-yl]oxy}aniline (420 mg, 992 μmol, intermediate 2) in pyridine (0.46 mL) and THF (6.8 mL) was slowly added at 0° C. phenyl carbonochloridate (140 μL, 1.1 mmol). After stirring this mixture for 5 minutes at 0° C. and then for 30 minutes at room temperature ethyl acetate (150 mL) was added. This organic phase was washed with 1N hydrochloric acid (30 mL), water, concentrated aqueous sodium hydrogencarbonate, brine, dried over sodium sulfate, filtered and concentrated to dryness. The resulting residue was purified via a Biotage chromatography system (100 g snap KP-NH column, hexane/10-70% ethyl acetate) to obtain 471 mg (79% purity, 69% yield) of the desired title compound.
1H-NMR (400 MHz, DMSO-d6) δ [ppm]: −0.11-−0.07 (m, 9H), 0.80-0.88 (m, 2H), 3.54-3.60 (m, 2H), 5.68 (s, 2H), 6.72-6.76 (m, 3H), 7.12-7.30 (m, 4H), 7.40-7.48 (m, 1H), 7.62 (d, 2H), 8.26 (d, 1H), 8.32 (s, 1H), 10.39 (br s, 1H).
To solution of phenyl (4-{[3-(trifluoromethyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}-1H-pyrrolo[2,3-b]pyridin-4-yl]oxy}phenyl)carbamate (150 mg, 276 μmol, intermediate 3) in DMF (1.3 mL) was added 2-(morpholin-4-yl)ethanamine (36 μL, 280 μmol) and this mixture was stirred at 60° C. for 2 hours. After cooling to room temperature ethyl acetate and water was added. After separation of the organic phase the aqueous phase was extracted two times with ethyl acetate. The combined organic phases were washed 3 times with half concentrated aqueous sodium chloride solution, dried over sodium sulfate, filtered and concentrated to dryness. The resulting residue was purified via a Biotage chromatography system (snap KP-NH column, hexane/50-100% ethyl acetate, then ethyl acetate/0-50% methanol) to obtain 63.8 mg (91% purity, 36% yield) of the desired title compound.
1H-NMR (400 MHz, DMSO-d6) δ [ppm]: −0.10-−0.07 (m, 9H), 0.80-0.88 (m, 2H), 2.35-2.43 (m, 6H), 3.18-3.26 (m, 2H), 3.52-3.63 (m, 6H), 5.67 (s, 2H), 6.45 (d, 1H), 7.06-7.17 (m, 3H), 7.47-7.53 (m, 2H), 8.24 (d, 1H), 8.30 (s, 1H), 8.87 (br s, 1H).
In analogy to intermediate 4), using phenyl (4-{[3-(trifluoromethyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}-1H-pyrrolo[2,3-b]pyridin-4-yl]oxy}phenyl)carbamate (150 mg, 276 μmol, intermediate 3) and 3-(morpholin-4-yl)propan-1-amine (40 μL, 280 μmol) in DMF (1.3 mL), we obtained after one single purification using a Biotage chromatography system 119 mg (93% purity, 68% yield) of the desired title compound.
1H-NMR (400 MHz, DMSO-d6) δ [ppm]: −0.11-−0.06 (m, 9H), 0.80-0.88 (m, 2H), 1.59 (quin, 2H), 2.25-2.40 (m, 6H), 3.12 (q, 2H), 3.53-3.60 (m, 6H), 5.67 (s, 2H), 6.17 (t, 1H), 6.45 (d, 1H), 7.06-7.11 (m, 2H), 7.47-7.52 (m, 2H), 8.24 (d, 1H), 8.30 (s, 1H), 8.58 (s, 1H).
In analogy to intermediate 1), using 6-chloro-4-nitro-3-(trifluoromethyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}-1H-pyrrolo[2,3-b]pyridine (500 mg, 1.26 mmol), 4-amino-2-fluorophenol (241 mg, 1.89 mmol) and (524 mg, 3.79 mmol) in DMSO (5.0 mL) we obtained after one single purification using a Biotage chromatography system 341 mg (88% purity, 50% yield) of the desired title compound.
1H-NMR (400 MHz, DMSO-d6) δ [ppm]: −0.10-−0.06 (m, 9H), 0.81-0.88 (m, 2H), 3.54-3.60 (m, 2H), 5.57 (s, 2H), 5.61 (s, 2H), 6.36 (d, 1H), 6.45 (ddd, 1H), 6.54 (dd, 1H), 7.07 (t, 1H), 8.35 (d, 1H).
In analogy to intermediate 2), using 4-{[6-chloro-3-(trifluoromethyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}-1H-pyrrolo[2,3-b]pyridin-4-yl]oxy}-3-fluoroaniline (335 mg, 704 μmol), intermediate 6), triethylamine (120 μL, 840 μmol) and 10% Pd on carbon (33.5 mg) in ethanol (24 mL), we obtained after one single purification using a Biotage chromatography system 240 mg (92% purity, 66% yield) of the desired title compound.
1H-NMR (400 MHz, DMSO-d6) δ [ppm]: −0.11-−0.07 (m, 9H), 0.77-0.89 (m, 2H), 3.53-3.60 (m, 2H), 5.49 (s, 2H), 5.66 (s, 2H), 6.40-6.47 (m, 2H), 6.52 (dd, 1H), 7.03 (t, 1H), 8.24 (d, 1H), 8.29 (s, 1H).
In analogy to intermediate 3), using 3-fluoro-4-{[3-(trifluoromethyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}-1H-pyrrolo[2,3-b]pyridin-4-yl]oxy}aniline (235 mg, 532 μmol), intermediate 7) and phenyl carbonochloridate (73 μL, 590 μmol) in pyridine (240 μL, 3.0 mmol) and THF (3.7 mL), we obtained after one single purification using a Biotage chromatography system 83.8 mg (72% purity, 20% yield) of the desired title compound.
1H-NMR (400 MHz, DMSO-d6) δ [ppm]: −0.10-−0.08 (m, 9H), 0.80-0.87 (m, 2H), 3.54-3.60 (m, 2H), 5.68 (s, 2H), 6.49 (dd, 1H), 6.72-6.78 (m, 2H), 7.12-7.31 (m, 4H), 7.37-7.48 (m, 1H), 7.65 (dd, 1H), 8.27 (d, 1H), 8.35 (s, 1H), 10.62 (br s, 1H).
In analogy to intermediate 4), using phenyl (3-fluoro-4-{[3-(trifluoromethyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}-1H-pyrrolo[2,3-b]pyridin-4-yl]oxy}phenyl)carbamate (80.0 mg, 142 μmol), intermediate 8) and 3-(morpholin-4-yl)propan-1-amine (21 μL, 140 μmol) in DMF (700 μL), we obtained after one single purification using a Biotage chromatography system 58.9 mg (93% purity, 63% yield) of the desired title compound.
1H-NMR (400 MHz, DMSO-d6) δ [ppm]: −0.10-−0.08 (m, 9H), 0.80-0.87 (m, 2H), 1.60 (quin, 2H), 2.27-2.37 (m, 6H), 3.09-3.16 (m, 2H), 3.54-3.61 (m, 6H), 5.68 (s, 2H), 6.28 (t, 1H), 6.45 (dd, 1H), 7.12-7.18 (m, 1H), 7.27 (t, 1H), 7.68 (dd, 1H), 8.26 (d, 1H), 8.33 (s, 1H), 8.81 (s, 1H).
In analogy to intermediate 1), using 6-chloro-4-nitro-3-(trifluoromethyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}-1H-pyrrolo[2,3-b]pyridine (750 mg, 1.89 mmol), 4-amino-2-methylphenol (257 mg, 2.08 mmol) and potassium carbonate (786 mg, 5.68 mmol) in DMSO (7.5 mL), we obtained after one single purification using a Biotage chromatography system 589 mg (96% purity, 63% yield) of the desired title compound.
1H-NMR (400 MHz, DMSO-d6) δ [ppm]: −0.11-−0.08 (m, 9H), 0.80-0.88 (m, 2H), 1.93 (s, 3H), 3.54-3.61 (m, 2H), 5.16 (s, 2H), 5.61 (s, 2H), 6.20 (s, 1H), 6.50 (dd, 1H), 6.54 (d, 1H), 6.87 (d, 1H), 8.33 (s, 1H).
In analogy to intermediate 2), using 4-{[6-chloro-3-(trifluoromethyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}-1H-pyrrolo[2,3-b]pyridin-4-yl]oxy}-3-methylaniline (585 mg, 1.24 mmol), intermediate 10), triethylamine (210 μL, 1.5 mmol) and 10% Pd on carbon (58.5 mg) in ethanol (43 mL), we obtained after one single purification using a Biotage chromatography system 468 mg (90% purity, 89% yield) of the desired title compound.
1H-NMR (400 MHz, DMSO-d6) δ [ppm]: −0.11-−0.09 (m, 9H), 0.78-0.86 (m, 2H), 1.92 (s, 3H), 3.54-3.60 (m, 2H), 5.08 (s, 2H), 5.66 (s, 2H), 6.28 (d, 1H), 6.48 (dd, 1H), 6.53 (d, 1H), 6.82 (d, 1H), 8.19 (d, 1H), 8.26 (s, 1H).
In analogy to intermediate 3), using 3-methyl-4-{[3-(trifluoromethyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}-1H-pyrrolo[2,3-b]pyridin-4-yl]oxy}aniline (463 mg, 1.06 mmol), intermediate 11) and phenyl carbonochloridate (150 μL, 1.2 mmol) in pyridine (490 μL, 6.1 mmol) and THF (7.3 mL), we obtained after one single purification using a Biotage chromatography system 272 mg (81% purity, 37% yield) of the desired title compound.
1H-NMR (400 MHz, DMSO-d6) δ [ppm]: −0.11-−0.08 (m, 9H), 0.80-0.86 (m, 2H), 1.99 (s, 3H), 3.55-3.61 (m, 2H), 5.67 (s, 2H), 6.32 (d, 1H), 6.72-6.78 (m, 2H), 7.12-7.30 (m, 3H), 7.41-7.48 (m, 1H), 7.54 (d, 1H), 8.23 (d, 1H), 8.31 (s, 1H), 9.33 (s, 1H), 10.34 (br s, 1H).
In analogy to intermediate 4), using phenyl (3-methyl-4-{[3-(trifluoromethyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}-1H-pyrrolo[2,3-b]pyridin-4-yl]oxy}phenyl)carbamate (135 mg, 242 μmol, intermediate 12) and 3-(morpholin-4-yl)propan-1-amine (35 μL, 240 μmol) in DMF (1.2 mL) we obtained after one single purification using a Biotage chromatography system 91.8 mg (95% purity, 62% yield) of the desired title compound.
1H-NMR (400 MHz, DMSO-d6) δ [ppm]: −0.12-−0.08 (m, 9H), 0.79-0.86 (m, 2H), 1.59 (quin, 2H), 2.02 (s, 3H), 2.27-2.38 (m, 6H), 3.12 (q, 2H), 3.55-3.61 (m, 6H), 5.67 (s, 2H), 6.17 (t, 1H), 6.29 (d, 1H), 7.04 (d, 1H), 7.33 (dd, 1H), 7.41 (d, 1H), 8.21 (d, 1H), 8.29 (s, 1H), 8.52 (s, 1H).
In analogy to intermediate 4), using phenyl (3-methyl-4-{[3-(trifluoromethyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}-1H-pyrrolo[2,3-b]pyridin-4-yl]oxy}phenyl)carbamate (112 mg, 201 μmol, intermediate 12) and 2-(morpholin-4-yl)ethanamine (26 μL, 200 μmol) in DMF (1.0 mL) we obtained after one single purification using a Biotage chromatography system 74.1 mg (100% purity, 62% yield) of the desired title compound.
1H-NMR (400 MHz, DMSO-d6) δ [ppm]: −0.12-−0.08 (m, 9H), 0.79-0.86 (m, 2H), 2.02 (s, 3H), 2.35-2.45 (m, 6H), 3.22 (q, 2H), 3.55-3.63 (m, 6H), 5.67 (s, 2H), 6.11 (t, 1H), 6.29 (d, 1H), 7.04 (d, 1H), 7.33 (dd, 1H), 7.41 (d, 1H), 8.21 (d, 1H), 8.29 (s, 1H), 8.71 (s, 1H).
In analogy to intermediate 1), using 6-chloro-4-nitro-3-(trifluoromethyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}-1H-pyrrolo[2,3-b]pyridine (500 mg, 1.26 mmol), 5-amino-2-hydroxybenzonitrile (186 mg, 1.39 mmol) and potassium carbonate (524 mg, 3.79 mmol) in DMSO (5.0 mL), we obtained after one single purification using a Biotage chromatography system 477 mg (92% purity, 72% yield) of the desired title compound.
1H-NMR (400 MHz, DMSO-d6) δ [ppm]: −0.09-−0.06 (m, 9H), 0.81-0.89 (m, 2H), 3.55-3.61 (m, 2H), 5.63 (s, 2H), 5.75 (s, 2H), 6.41 (s, 1H), 6.94-6.99 (m, 2H), 7.19-7.23 (m, 1H), 8.39 (s, 1H).
In analogy to intermediate 2), using 5-amino-2-{[6-chloro-3-(trifluoromethyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}-1H-pyrrolo[2,3-b]pyridin-4-yl]oxy}benzonitrile (473 mg, 0.98 mmol), intermediate 15), triethylamine (164 μL, 1.2 mmol) and 10% Pd on carbon (71.6 mg) in ethanol (34 mL), we obtained after one single purification using a Biotage chromatography system 321 mg (93% purity, 68% yield) of the desired title compound.
1H-NMR (400 MHz, DMSO-d6) δ [ppm]: −0.10-−0.07 (m, 9H), 0.81-0.87 (m, 2H), 3.55-3.60 (m, 2H), 5.68 (s, 2H), 5.68 (s, 2H), 6.43 (d, 1H), 6.93-6.98 (m, 2H), 7.12-7.17 (m, 1H), 8.27 (d, 1H), 8.33 (s, 1H).
In analogy to intermediate 3), using 5-amino-2-{[3-(trifluoromethyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}-1H-pyrrolo[2,3-b]pyridin-4-yl]oxy}benzonitrile (317 mg, 707 μmol), intermediate 16) and phenyl carbonochloridate (98 μL, 770 μmol) in pyridine (325 μL, 4.0 mmol) and THF (4.8 mL), we obtained after one single purification using a Biotage chromatography system 329 mg (94% purity, 77% yield) of the desired title compound.
1H-NMR (400 MHz, DMSO-d6) δ [ppm]: −0.10-−0.07 (m, 9H), 0.81-0.88 (m, 2H), 3.56-3.62 (m, 2H), 5.70 (s, 2H), 6.65 (d, 1H), 6.72-6.78 (m, 1H), 7.24-7.31 (m, 3H), 7.43-7.48 (m, 2H), 7.85 (dd, 1H), 8.03 (d, 1H), 8.34 (d, 1H), 8.39 (s, 1H), 10.71 (br s, 1H).
In analogy to intermediate 4), using phenyl (3-cyano-4-{[3-(trifluoromethyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}-1H-pyrrolo[2,3-b]pyridin-4-yl]oxy}phenyl)carbamate (164 mg, 289 μmol), intermediate 17) and 2-(morpholin-4-yl)ethanamine (49 μL, 380 μmol) in DMF (1.4 mL), we obtained after one single purification using a Biotage chromatography system 115 mg (93% purity, 61% yield) of the desired title compound.
1H-NMR (400 MHz, DMSO-d6) δ [ppm]: −0.10-−0.07 (m, 9H), 0.81-0.88 (m, 2H), 2.36-2.43 (m, 6H), 3.23 (q, 2H), 3.55-3.63 (m, 6H), 5.69 (s, 2H), 6.28 (t, 1H), 6.57 (d, 1H), 7.33 (d, 1H), 7.67 (dd, 1H), 8.04 (d, 1H), 8.32 (d, 1H), 8.37 (s, 1H), 9.11 (s, 1H).
In analogy to intermediate 4), using phenyl (3-cyano-4-{[3-(trifluoromethyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}-1H-pyrrolo[2,3-b]pyridin-4-yl]oxy}phenyl)carbamate (161 mg, 283 μmol), intermediate 17) and 3-(morpholin-4-yl)propan-1-amine (54 μL, 367 mol) in DMF (1.4 mL), we obtained after one single purification using a Biotage chromatography system 207 mg (84% purity, 99% yield) of the desired title compound.
1H-NMR (400 MHz, DMSO-d6) δ [ppm]: −0.11-−0.06 (m, 9H), 0.81-0.88 (m, 2H), 1.60 (quin, 2H), 2.27-2.38 (m, 6H), 3.13 (q, 2H), 3.54-3.62 (m, 6H), 5.69 (s, 2H), 6.38 (t, 1H), 6.56 (d, 1H), 7.32 (d, 1H), 7.69 (dd, 1H), 8.04 (d, 1H), 8.31 (d, 1H), 8.37 (s, 1H), 8.92 (s, 1H).
In analogy to intermediate 1), using 6-chloro-4-nitro-3-(trifluoromethyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}-1H-pyrrolo[2,3-b]pyridine (500 mg, 1.26 mmol), 4-amino2-chlorophenol (199 mg, 1.39 mmol) and potassium carbonate (524 mg, 3.79 mmol) in DMSO (5.0 mL), we obtained after one single purification using a Biotage chromatography system 467 mg (75% yield) of the desired title compound.
1H-NMR (400 MHz, DMSO-d6) δ [ppm]: −0.12-−0.06 (m, 9H), 0.81-0.87 (m, 2H), 3.54-3.60 (m, 2H), 5.56 (s, 2H), 5.61 (s, 2H), 6.23 (s, 1H), 6.62 (dd, 1H), 6.77 (d, 1H), 7.11 (d, 1H), 8.35 (s, 1H).
In analogy to intermediate 2), using 3-chloro-4-{[6-chloro-3-(trifluoromethyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}-1H-pyrrolo[2,3-b]pyridin-4-yl]oxy}aniline (463 mg, 940 μmol, intermediate 20), triethylamine (160 μL, 1.1 mmol) and 10% Pd on carbon (46.3 mg) in ethanol (33 mL), we obtained after one single purification using a Biotage chromatography system 418 mg (97% yield) of the desired title compound.
1H-NMR (400 MHz, DMSO-d6) δ [ppm]: −0.11-−0.08 (m, 9H), 0.79-0.86 (m, 2H), 3.53-3.60 (m, 2H), 5.48 (s, 2H), 5.66 (s, 2H), 6.30 (d, 1H), 6.61 (dd, 1H), 6.76 (d, 1H), 7.05 (d, 1H), 8.22 (d, 1H), 8.28 (s, 1H).
In analogy to intermediate 3), using 3-chloro-4-{[3-(trifluoromethyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}-1H-pyrrolo[2,3-b]pyridin-4-yl]oxy}aniline (414 mg, 904 μmol, intermediate 21) and phenyl carbonochloridate (120 μL, 980 μmol) in pyridine (420 μL, 5.1 mmol), we obtained after one single purification using a Biotage chromatography system 470 mg (70% purity, 63% yield) of the desired title compound.
1H-NMR (400 MHz, DMSO-d6) δ [ppm]: −0.11-−0.04 (m, 9H), 0.79-0.87 (m, 2H), 3.54-3.61 (m, 2H), 5.68 (s, 2H), 6.37 (d, 1H), 7.22-7.32 (m, 3H), 7.41-7.48 (m, 2H), 7.57 (dd, 1H), 7.63 (d, 1H), 7.84 (d, 1H), 8.26 (d, 1H), 8.34 (s, 1H), 10.60 (br s, 1H).
In analogy to intermediate 4), phenyl (3-chloro-4-{[3-(trifluoromethyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}-1H-pyrrolo[2,3-b]pyridin-4-yl]oxy}phenyl)carbamate (150 mg, 259 μmol, intermediate 22) and 3-(morpholin-4-yl)propan-1-amine (38 μL, 260 μmol) in DMF (1.3 mL), we obtained after one single purification using a Biotage chromatography system 160 mg (71% purity, 69% yield) of the desired title compound.
1H-NMR (400 MHz, DMSO-d6) δ [ppm]: −0.10-−0.05 (m, 9H), 0.80-0.87 (m, 2H), 1.60 (quin, 2H), 2.26-2.38 (m, 6H), 3.08-3.17 (m, 2H), 3.54-3.61 (m, 6H), 5.67 (s, 2H), 6.27-6.34 (m, 2H), 7.26-7.35 (m, 2H), 7.86 (d, 1H), 8.24 (d, 1H), 8.32 (s, 1H), 8.80 (s, 1H).
In analogy to intermediate 4), phenyl (3-chloro-4-{[3-(trifluoromethyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}-1H-pyrrolo[2,3-b]pyridin-4-yl]oxy}phenyl)carbamate (150 mg, 259 μmol, intermediate 22) and 2-(morpholin-4-yl)ethanamine (34 μL, 260 μmol) in DMF (1.3 mL), we obtained after one single purification using a Biotage chromatography system 153 mg (72% purity, 69% yield) of the desired title compound.
1H-NMR (400 MHz, DMSO-d6) δ [ppm]: −0.11-−0.05 (m, 9H), 0.79-0.88 (m, 2H), 2.36-2.43 (m, 6H), 3.19-3.27 (m, 2H), 3.55-3.63 (m, 6H), 5.67 (s, 2H), 6.20 (t, 1H), 6.33 (d, 1H), 7.27-7.34 (m, 2H), 7.87 (d, 1H), 8.24 (d, 1H), 8.32 (s, 1H), 8.99 (s, 1H).
To a solution of 1-[2-(morpholin-4-yl)ethyl]-3-(4-{[3-(trifluoromethyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}-1H-pyrrolo[2,3-b]pyridin-4-yl]oxy}phenyl)urea (60.0 mg, 104 μmol, intermediate 4) in dichloromethane (1.4 mL) was added trifluoroacidic acid (700 μL, 9.1 mmol) and this mixture was stirred for 3 hours at room temperature. The reaction mixture was carefully poured into an aqueous solution of sodium hydrogencarbonate. This aqueous phase was extracted two times with ethyl actetate. Then the combined organic phases were washed with brine, dried over sodium sulfate and then after filtration evaporated to dryness in vacuum. The obtained crude product was purified via a Biotage chromatography system (25 g snap KP-NH column, ethyl acetate/0-50% methanol) to obtain 35.3 mg (90% purity, 68% yield) of the desired title compound.
1H-NMR (400 MHz, DMSO-d6) δ [ppm]: 2.36-2.42 (m, 6H), 3.22 (q, 2H), 3.60 (t, 4H), 6.10 (t, 1H), 6.36 (d, 1H), 7.06-7.10 (m, 2H), 7.47-7.52 (m, 2H), 8.05 (s, 1H), 8.17 (d, 1H), 8.77 (s, 1H), 12.50 (s, 1H).
In analogy to example 1), 1-[3-(morpholin-4-yl)propyl]-3-(4-{[3-(trifluoromethyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}-1H-pyrrolo[2,3-b]pyridin-4-yl]oxy}phenyl)urea (115 mg, 194 μmol, intermediate 5) was stirred with trifluoroacidic acid (1.3 mL, 17 mmol) in dichloromethane (2.6 mL) to obtain 71.1 mg (97% purity, 77% yield) of the desired title compound.
1H-NMR (400 MHz, DMSO-d6) δ [ppm]: 1.59 (quin, 2H), 2.27-2.38 (m, 6H), 3.12 (q, 2H), 3.57 (t, 4H), 6.16 (t, 1H), 6.36 (d, 1H), 7.05-7.11 (m, 2H), 7.46-7.51 (m, 2H), 8.05 (s, 1H), 8.17 (d, 1H), 8.57 (s, 1H), 12.50 (s, 1H).
In analogy to example 1), 1-(3-fluoro-4-{[3-(trifluoromethyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}-1H-pyrrolo[2,3-b]pyridin-4-yl]oxy}phenyl)-3-[3-(morpholin-4-yl)propyl]urea (56.0 mg, 91.5 μmol, intermediate 9) was stirred with trifluoroacidic acid (630 μL, 8.1 mmol) in dichloromethane (1.4 mL) to obtain 33.0 mg (94% purity, 70% yield) of the desired title compound.
1H-NMR (400 MHz, DMSO-d6) δ [ppm]: 1.60 (quin, 2H), 2.27-2.37 (m, 6H), 3.12 (q, 2H), 3.58 (t, 4H), 6.29 (t, 1H), 6.36 (dd, 1H), 7.10-7.16 (m, 1H), 7.25 (t, 1H), 7.69 (dd, 1H), 8.08 (s, 1H), 8.18 (d, 1H), 8.81 (s, 1H), 12.56 (br s, 1H).
In analogy to example 1), 1-(3-methyl-4-{[3-(trifluoromethyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}-1H-pyrrolo[2,3-b]pyridin-4-yl]oxy}phenyl)-3-[3-(morpholin-4-yl)propyl]urea (88.0 mg, 145 μmol, intermediate 13) was stirred with trifluoroacidic acid (1.0 mL, 13 mmol) in dichloromethane (2.0 mL) to obtain 53.4 mg (95% purity, 73% yield) of the desired title compound.
1H-NMR (400 MHz, DMSO-d6) δ [ppm]: 1.59 (quin, 2H), 2.03 (s, 3H), 2.27-2.38 (m, 6H), 3.12 (q, 2H), 3.58 (t, 4H), 6.16 (t, 1H), 6.20 (d, 1H), 7.02 (d, 1H), 7.32 (dd, 1H), 7.40 (d, 1H), 8.04 (s, 1H), 8.14 (d, 1H), 8.50 (s, 1H), 12.49 (s, 1H).
In analogy to example 1), 1-(3-methyl-4-{[3-(trifluoromethyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}-1H-pyrrolo[2,3-b]pyridin-4-yl]oxy}phenyl)-3-[2-(morpholin-4-yl)ethyl]urea (70.0 mg, 118 μmol, intermediate 14) was stirred with trifluoroacidic acid (800 μL, 10 mmol) in dichloromethane (1.6 mL) to obtain 45.4 mg (95% purity, 79% yield) of the desired title compound.
1H-NMR (400 MHz, DMSO-d6) δ [ppm]: 2.03 (s, 3H), 2.36-2.44 (m, 6H), 3.22 (q, 2H), 3.60 (t, 4H), 6.10 (t, 1H), 6.20 (d, 1H), 7.02 (d, 1H), 7.32 (dd, 1H), 7.41 (d, 1H), 8.04 (s, 1H), 8.14 (d, 1H), 8.70 (s, 1H), 12.49 (s, 1H).
In analogy to example 1), 1-(3-cyano-4-{[3-(trifluoromethyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}-1H-pyrrolo[2,3-b]pyridin-4-yl]oxy}phenyl)-3-[2-(morpholin-4-yl)ethyl]urea (110 mg, 182 μmol, intermediate 18) was stirred with trifluoroacidic acid (1.2 mL, 16 mmol) in dichloromethane (2.4 mL) to obtain 85.3 mg (95% purity, 90% yield) of the desired title compound.
1H-NMR (500 MHz, DMSO-d6) δ [ppm]: 2.37-2.44 (m, 6H), 3.23 (q, 2H), 3.59 (t, 4H), 6.27 (t, 1H), 6.48 (d, 1H), 7.30 (d, 1H), 7.66 (dd, 1H), 8.04 (d, 1H), 8.13 (s, 1H), 8.24 (d, 1H), 9.09 (s, 1H), 12.65 (s, 1H).
In analogy to example 1), 1-(3-cyano-4-{[3-(trifluoromethyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}-1H-pyrrolo[2,3-b]pyridin-4-yl]oxy}phenyl)-3-[3-(morpholin-4-yl)propyl]urea (203 mg, 328 μmol, intermediate 19) was stirred with trifluoroacidic acid (2.2 mL, 29 mmol) in dichloromethane (4.4 mL) to obtain 103.3 mg (95% purity, 61% yield) of the desired title compound.
1H-NMR (500 MHz, DMSO-d6) δ [ppm]: 1.60 (quin, 2H), 2.27-2.37 (m, 6H), 3.13 (q, 2H), 3.57 (t, 4H), 6.41 (t, 1H), 6.47 (d, 1H), 7.29 (d, 1H), 7.67 (dd, 1H), 8.03 (d, 1H), 8.12 (s, 1H), 8.24 (d, 1H), 8.95 (s, 1H), 12.65 (br s, 1H).
In analogy to example 1), 1-(3-chloro-4-{[3-(trifluoromethyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}-1H-pyrrolo[2,3-b]pyridin-4-yl]oxy}phenyl)-3-[3-(morpholin-4-yl)propyl]urea (155 mg, 247 μmol, intermediate 23) was stirred with trifluoroacidic acid (1.7 mL, 22 mmol) in dichloromethane (3.4 mL) to obtain after an additional HPLC purification (method 1) 21.7 mg (95% purity, 17% yield) of the desired title compound.
1H-NMR (400 MHz, DMSO-d6) δ [ppm]: 1.60 (quin, 2H), 2.27-2.38 (m, 6H), 3.13 (q, 2H), 3.58 (t, 4H), 6.24 (d, 1H), 6.28 (t, 1H), 7.26 (d, 1H), 7.32 (dd, 1H), 7.86 (d, 1H), 8.07 (d, 1H), 8.17 (d, 1H), 8.77 (s, 1H), 12.55 (d, 1H).
In analogy to example 1), 1-(3-chloro-4-{[3-(trifluoromethyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}-1H-pyrrolo[2,3-b]pyridin-4-yl]oxy}phenyl)-3-[2-(morpholin-4-yl)ethyl]urea (147 mg, 239 μmol, intermediate 24) was stirred with trifluoroacidic acid (1.7 mL, 22 mmol) in dichloromethane (3.3 mL) to obtain after an additional HPLC purification (method 1) 19.1 mg (95% purity, 16% yield) of the desired title compound.
1H-NMR (400 MHz, DMSO-d6) δ [ppm]: 2.36-2.43 (m, 6H), 3.23 (q, 2H), 3.60 (t, 4H), 6.19 (t, 1H), 6.24 (d, 1H), 7.26 (d, 1H), 7.31 (dd, 1H), 7.86 (d, 1H), 8.07 (d, 1H), 8.17 (d, 1H), 8.97 (s, 1H), 12.54 (br s, 1H).
The in vitro activity of the compounds of the present invention can be demonstrated in the following assays:
The example testing experiments described herein serve to illustrate the present invention and the invention is not limited to the examples given.
In order that this invention may be better understood, the following examples are set forth.
These examples are for the purpose of illustration only, and are not to be construed as limiting the scope of the invention in any manner. All publications mentioned herein are incorporated by reference in their entirety.
Demonstration of the activity of the compounds of the present invention may be accomplished through in vitro and in vivo assays that are well known in the art. For example, to demonstrate the efficacy of a pharmaceutical agent to inhibit and be selective against e.g. TBK1 the following assays may be used.
The ability of the compounds of the present invention to inhibit the binding of an Alexa647-labelled ATP-competitive kinase inhibitor to a Glutathione-S-transferase- (GST-) fusion protein was quantified employing the TR-FRET-based binding competition assay as described in the following paragraphs.
A recombinant fusion protein of N-terminal GST and full-length human, expressed by baculovirus infected SF9 insect cells and purified by Glutathione Sepharose affinity chromatography, was used as GST-fusion protein. Tracer 222 from Invitrogen (catalogue no. PR9198A) was used as Alexa647-labelled ATP-competitive kinase inhibitor. For the assay 50 nl of a 100 fold concentrated solution of the test compound in DMSO was pipetted into either a black low volume 384 well microtiter plate or a black 1536 well microtiter plate (both Greiner Bio-One, Frickenhausen, Germany), 3 μl solution of Tracer 222 (25 nM=>final concentration in 5 μl assay volume is 15 nM) in aqueous assay buffer [25 mM Tris/HCl pH 7.5, 10 mM MgCl2, 5 mM β-glycerolphosphate, 2.5 mM dithiothreitol, 0.5 mM ethylene glycol-bis(2-aminoethylether)-N,N,N′,N′-tetraacetic acid [EGTA], 0.5 mM sodium ortho-vanadate, 0.01% (w/v) bovine serum albumin [BSA], 0.005% (w/v) Pluronic F-127 (Sigma)] were added. Then the binding competition was started by the addition of 2 μl of a solution of the GST-fusion protein (2.5 nM=>final conc. in the 5 μl assay volume is 1 nM) and of Anti-GST-Tb (1.25 nM=>final conc. in the 5 μl assay volume is 0.5 nM), a Lumi4®-Tb Cryptate-conjugated anti-GST-antibody from Cisbio Bioassays (France), in assay buffer.
The resulting mixture was incubated 30 min at 22° C. to allow the formation of a complex between the Tracer 222, the fusion protein and Anti-GST-Tb. Subsequently the amount of this complex was evaluated by measurement of the resonance energy transfer from the Tb-cryptate to the Tracer 222. Therefore, the fluorescence emissions at 620 nm and 665 nm after excitation at 350 nm were measured in a TR-FRET reader, e.g. a Pherastar (BMG Labtechnologies, Offenburg, Germany) or a Viewlux (Perkin-Elmer). The ratio of the emissions at 665 nm and at 622 nm was taken as the measure for the amount of the complex. The data were normalised (assay reaction without inhibitor=0% inhibition, all other assay components but GST-fusion protein=100% inhibition). Usually the test compounds were tested on the same microtiterplate in 11 different concentrations in the range of 20 μM to 0.07 nM (20 μM, 5.7 μM, 1.6 μM, 0.47 μM, 0.13 μM, 38 nM, 11 nM, 3.1 nM, 0.9 nM, 0.25 nM and 0.07 nM, the dilution series prepared separately before the assay on the level of the 100 fold concentrated solutions in DMSO by serial dilutions, exact concentrations may vary depending pipettors used) in duplicate values for each concentration and IC50 values were calculated using Genedata Screener™ software.
TBK1-inhibitory activity of compounds of the present invention at a high ATP concentration after preincubation of enzyme and test compounds was quantified employing the TR-FRET-based TBK1 assay as described in the following paragraphs.
Recombinant full-length N-terminally His-tagged human TBK1, expressed in insect cells and purified by Ni-NTA affinity chromatography, was purchased from Life Technologies (Cat. No PR5618B) and used as enzyme. As substrate for the kinase reaction biotinylated peptide biotin-Ahx-GDEDFSSFAEPG (C-terminus in amide form) was used which can be purchased e.g. form the company Biosyntan (Berlin-Buch, Germany).
For the assay 50 nl of a 100 fold concentrated solution of the test compound in DMSO was pipetted into either a black low volume 384 well microtiter plate or a black 1536 well microtiter plate (both Greiner Bio-One, Frickenhausen, Germany), 2 μl of a solution of TBK1 in aqueous assay buffer [50 mM HEPES pH 7.0, 10 mM MgCl2, 1.0 mM dithiothreitol, 0.05% (w/v) bovine serum albumine, 0.01% (v/v) Nonidet-P40 (Sigma), protease inhibitor mixture (“Complete w/o EDTA” from Roche, 1 tablet per 5 mL)] were added and the mixture was incubated for 15 min at 22° C. to allow pre-binding of the test compounds to the enzyme before the start of the kinase reaction. Then the kinase reaction was started by the addition of 3 μl of a solution of adenosine-tri-phosphate (ATP, 1.67 mM=>final conc. in the 5 μl assay volume is 1 mM) and substrate (1.67 μM=>final conc. in the 5 μl assay volume is 1 μM) in assay buffer and the resulting mixture was incubated for a reaction time of 30 min at 22° C. The concentration of TBK1 was adjusted depending of the activity of the enzyme lot and was chosen appropriate to have the assay in the linear range, typical concentrations were in the range of 0.002-0.004 μg/mL. The reaction was stopped by the addition of 3 μl of a solution of TR-FRET detection reagents (0.33 μM streptavidine-XL665 [Cisbio Bioassays, Codolet, France], 2.5 nM anti-phosho-Serine antibody [Merck Millipore, “STK antibody”, cat. #35-002] and 1.25 nM LANCE EU-W1024 labeled anti-mouse IgG antibody [Perkin-Elmer, product no. AD0077]) in an aqueous EDTA-solution (167 mM EDTA, 0.13% (w/v) bovine serum albumin in 100 mM HEPES/NaOH pH 7.5).
The resulting mixture was incubated 1 h at 22° C. to allow the formation of complex between the phosphorylated biotinylated peptide and the detection reagents. Subsequently the amount of phosphorylated substrate was evaluated by measurement of the resonance energy transfer from the Eu-chelate to the streptavidine-XL. Therefore, the fluorescence emissions at 620 nm and 665 nm after excitation at 350 nm was measured in a TR-FRET reader, e.g. a Pherastar (BMG Labtechnologies, Offenburg, Germany) or a Viewlux (Perkin-Elmer). The ratio of the emissions at 665 nm and at 622 nm was taken as the measure for the amount of phosphorylated substrate. The data were normalised (enzyme reaction without inhibitor=0% inhibition, all other assay components but no enzyme=100% inhibition). Usually the test compounds were tested on the same microtiterplate in 11 different concentrations in the range of 20 μM to 0.07 nM (20 μM, 5.7 μM, 1.6 μM, 0.47 μM, 0.13 μM, 38 nM, 11 nM, 3.1 nM, 0.9 nM, 0.25 nM and 0.07 nM, the dilution series prepared separately before the assay on the level of the 100 fold concentrated solutions in DMSO by serial dilutions, exact concentrations may vary depending pipettors used) in duplicate values for each concentration and IC50 values were calculated using Genedata Screener™ software.
Phosphorylation assays were carried out in Jurkat E6.1 cells from American Type Culture Collection (ATCC) stably overexpressing human FLAG-tagged SLP-76 (proprietary). Cultured cells were kept in RPMI 1640 medium supplemented with 1% FCS at a cell density of 2×10e6/mL 24 h prior compound testing. Starved cells were simultaneously treated with 350 ng/mL a-CD3 antibody (clone OKT3. ebioscience #16-0037-85. plate-bound) and test compound for 30 min at 37° C. Applied compounds were tested at either fixed concentration of 10 μmol/L and 20 μmol/L or in a 8 point dose response titration of increase compound concentration with 10 nmol/L. 50 nmol/L. 100 nmol/L. 500 nmol/L. 1 μmol/L. 5 μmol/L. 10 μmol/L and 20 μmol/L in triplicates. The cells were washed once in phosphate-buffered saline (pH 7.4). Cells were lysed using a lysis buffer containing 50 mmol/L Tris-Cl (pH 7.5). 150 mM NaCl. 2 mM EDTA. 1% Triton-X 100. 0.5% Na-DOC. 0.1% SDS. 1/10 complete mini protease inhibitor cocktail (Roche #11836170001) and 1/10 PhosSTOP phosphatase inhibitor cocktail (Roche #04906837001). A total of 1.25 μg cell lysate was analyzed by capillary electrophoresis using the Peggy Sue™ System (Proteinsimple® San Jose, Calif. USA) with a 12-230 kDa size-based master kit with split buffer/a-rabbit-HRP #PS-MK18/a-mouse-HRP #PS-MK19 according to manufacture's protocol. Probe antibodies used were a rabbit monoclonal antibody supernatant raised against human phospho-Ser376-SLP-76 peptide (proprietary) and for mormalization an a-alpha-Tubulin. mouse monoclonal antibody (Sigma #T9026). As control for maximal effect (max control. which represent the maximally possible inhibition of pSer376-SLP-76 by a test compound) cells with no a-CD3 (clone OKT3. ebioscience #16-0037-85. plate-bound) and no test compound treatment were used. Cells with a-CD3 treatment only were used as negative control (min control. which represent the minimally possible inhibition of pSer376-SLP-76 by a test compound)
AUC values of each respective test sample were normalized using the AUC of housekeeping gene alpha-Tubulin and AUC of pSer376-SLP-76 of the min control. The percentage of the amount of pSer-SLP-76 in the treatment samples was calculated using the max control and min control values of the respective Peggy Sue™ run.
| Number | Date | Country | Kind |
|---|---|---|---|
| 17175806.3 | Jun 2017 | EP | regional |
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/EP2018/065045 | 6/7/2018 | WO | 00 |
