Patent Application
20240294542
The present invention claims priority to the following invention patent applications:
The present invention belongs to the technical field of medicines, and particularly relates to an NLRP3 inflammasome inhibitor and use thereof.
Nucleotide-binding oligomerization domain (NOD)-like receptor protein 3 (NLRP3) belongs to the NOD-like receptors (NLRs) family, also known as “pyrin domain-containing protein 3”. NLRP3 contains three modules which are a pyrin domain (PYD), a nucleotide-binding site domain (NBD), and a leucine-rich repeat (LRR). Upon receiving a stimulus from a sterile inflammatory risk signal, NLRP3 interacts with adaptor apoptosis-associated speck-like protein containing a CARD (ASC) and pro-caspase 1 to form an NLRP3 inflammasome. Activation of the NLRP3 inflammasome results in the release of interleukin-1β (IL-1β) and interleukin-18 (IL-18).
Activation of NLRP3 inflammasome usually requires two steps. The first step involves a priming signal, where Toll-like receptors recognize pathogen-associated molecular patterns (PAMPs) or damage-associated molecular patterns (DAMPs) which in turn transmit a signal into the cells to mediate the activation of the NF-κB signaling pathway, thereby up-regulating the transcription levels of NLRP3 inflammasome-associated components including inactive NLRP3 and pro-IL-1β. The second step involves an activation signal; after a P2X7 receptor and the like receive signal stimulation of ATP, nigericin and the like, an NLRP3 monomer is oligomerized to form an NLRP3 oligomer, and then ASC and pro-caspase 1 are recruited to assemble into an NLRP3 inflammasome complex. This triggers the conversion of pro-caspase 1 to caspase 1, and also the production and secretion of mature IL-1β and IL-1β.
Activation of NLRP3 inflammasome is associated with various diseases, for example, autoinflammatory fever syndromes such as cryo-pyrin-associated periodic syndromes (CAPS), sickle cell disease, systemic lupus erythematosus (SLE), chronic liver disease, nonalcoholic steatohepatitis (NASH), gout, pseudogout (chondrocalcinosis), type I and type II diabetes and related complications (e.g., nephropathy, retinopathy), neuroinflammation-related disorders (e.g., multiple sclerosis, brain infection, acute injury, neurodegenerative disease, Alzheimer's disease), atherosclerosis and cardiovascular risk (e.g., hypertension), hidradenitis suppurativa, wound healing and scarring, and cancer (e.g., colorectal cancer, lung cancer, myeloproliferative tumors, leukemia, myelodysplastic syndrome (MDS), myelofibrosis). Most treatment methods include symptomatic treatment, slowing of the progression of the diseases/disorders, and surgeries as the last treatment means.
Currently, as the varieties of NLRP3 inflammasome inhibitors are few, the development of an NLRP3 inflammasome inhibitor with relatively high activity and better druggability becomes clinical requirements.
The present invention researches the following compound, or a pharmaceutically acceptable salt, a stereoisomer or a tautomer thereof, and discovers that the compound, or the pharmaceutically acceptable salt, the stereoisomer or the tautomer thereof has relatively high biological activity on an NLRP3 inflammasome and has important clinical development value for treating NLRP3-associated diseases.
In order to achieve the above objective, the present invention provides the following technical solutions:
The technical solutions of the present invention include a compound of general formula (I), or a pharmaceutically acceptable salt, a stereoisomer or a tautomer thereof:
On the basis of the above technical solutions, the compound, or the pharmaceutically acceptable salt, the stereoisomer or the tautomer thereof provided in the present invention has a structure of general formula (II):
On the basis of the above technical solutions, for the compound, or the pharmaceutically acceptable salt, the stereoisomer or the tautomer thereof provided in the present invention, wherein ring A is selected from phenyl and 5-7 membered heteroaryl; ring A is optionally substituted with 1-4 substituents selected from hydroxyl, amino, carboxyl, cyano, nitro, halogen, C1-6 alkyl, —NH—C1-6 alkyl, haloC1-6 alkyl, C1-6 alkoxy, haloC1-6 alkoxy, 3-7 membered heterocyclyl, 3-7 membered cycloalkyl, C1-6 alkylsulfonyl, and —N(C1-6 alkyl)2.
On the basis of the above technical solutions, for the compound, or the pharmaceutically acceptable salt, the stereoisomer or the tautomer thereof provided in the present invention, wherein ring A is selected from,
ring A is optionally substituted with 1-4 substituents selected from hydroxyl, amino, carboxyl, cyano, nitro, halogen, C1-6 alkyl, —NH—C1-6 alkyl, haloC1-6 alkyl, C1-6 alkoxy, haloC1-6 alkoxy, 3-7 membered heterocyclyl, 3-7 membered cycloalkyl, C1-6 alkylsulfonyl, and —N(C1-6 alkyl)2.
On the basis of the above technical solutions, for the compound, or the pharmaceutically acceptable salt, the stereoisomer or the tautomer thereof provided in the present invention, wherein Y is selected from phenyl, 5-7 membered heteroaryl, 3-8 membered heterocyclyl, and 3-7 membered cycloalkyl; Y is optionally substituted with 1-3 substituents selected from halogen, cyano, amino, hydroxyl, carbonyl, C1-6 alkyl, haloC1-6 alkyl, C1-6 alkoxy, 3-7 membered heterocyclyl, 3-7 membered cycloalkyl, C1-6 alkylamino, C1-6 alkylcarbonylamino, C1-6 alkylsulfonyl, aminocarbonyl, C1-6 alkylaminocarbonyl, and sulfonyl; the C1-6 alkyl, haloC1-6 alkyl, C1-6 alkoxy, 3-7-membered heterocyclyl, 3-7-membered cycloalkyl and sulfonyl are optionally substituted by 1-3 substituents selected from halogen, cyano, amino, hydroxyl, carbonyl, and C1-6 alkyl.
On the basis of the above technical solutions, for the compound, or the pharmaceutically acceptable salt, the stereoisomer or the tautomer thereof provided in the present invention, wherein Y is selected from naphthyl, 8-14 membered fused heteroaryl, 6-12 membered fused heterocyclyl, and 6-12 membered fused cycloalkyl; Y is optionally substituted with 1-3 substituents selected from halogen, cyano, amino, hydroxyl, carbonyl, C1-6 alkyl, haloC1-6 alkyl, and C1-6 alkoxy.
On the basis of the above technical solutions, for the compound, or the pharmaceutically acceptable salt, the stereoisomer or the tautomer thereof provided in the present invention, wherein Y is substituted with cyano and is optionally substituted with 1-2 substituents selected from halogen, cyano, amino, hydroxyl, carbonyl, C1-6 alkyl, haloC1-6 alkyl, C1-6 alkoxy, 3-7-membered heterocyclyl, 3-7-membered cycloalkyl, C1-6 alkylamino, C1-6 alkylcarbonylamino, C1-6 alkylsulfonyl, aminocarbonyl, C1-6 alkylaminocarbonyl, and sulfonyl; the C1-6 alkyl, haloC1-6 alkyl, C1-6 alkoxy, 3-7-membered heterocyclyl, 3-7-membered cycloalkyl and sulfonyl are optionally substituted with 1-3 substituents selected from halogen, cyano, amino, hydroxyl, carbonyl, and C1-6 alkyl.
On the basis of the above technical solutions, for the compound, or the pharmaceutically acceptable salt, the stereoisomer or the tautomer thereof provided in the present invention, wherein R3 is selected from —NR4R5, —NH—COR5, and —O—R5; R4 is selected from hydrogen and C1-3 alkyl; R5 is selected from 3-7 membered cycloalkyl and 3-7 membered heterocyclyl, and R5 is optionally substituted with 1-2 substituents selected from C1-6 alkyl, hydroxyl, hydroxyl-substituted C1-6 alkyl, 3-7 membered cycloalkyl-substituted C1-6 alkyl, hydroxyl, halogen, C2-6 alkenylcarbonyl, C1-6 alkylsulfonyl, 3-7 membered cycloalkylsulfonyl, aminosulfonyl, 3-7 membered heterocyclyl, 3-7 membered cycloalkyl, carboxyl, and C1-6 alkylcarbonyl.
On the basis of the above technical solutions, for the compound, or the pharmaceutically acceptable salt, the stereoisomer or the tautomer thereof provided in the present invention, wherein R3 is —NH—R5, and R5 is 3-7 membered heterocyclyl substituted with 1-2 substituents selected from C1-6 alkyl.
On the basis of the above technical solutions, for the compound, or the pharmaceutically acceptable salt, the stereoisomer or the tautomer thereof provided in the present invention, wherein Y is selected from phenyl, 5-7 membered heteroaryl, 3-8 membered heterocyclyl, and 3-7 membered cycloalkyl; Y is optionally substituted with 1-3 substituents selected from halogen, cyano, amino, hydroxyl, carbonyl, C1-6 alkyl, haloC1-6 alkyl, C1-6 alkoxy, 3-7 membered heterocyclyl, 3-7 membered cycloalkyl, C1-6 alkylamino, C1-6 alkylcarbonylamino, C1-6 alkylsulfonyl, aminocarbonyl, C1-6 alkylaminocarbonyl, and sulfonyl; the C1-6 alkyl, haloC1-6 alkyl, C1-6 alkoxy, 3-7 membered heterocyclyl, 3-7 membered cycloalkyl and sulfonyl are optionally substituted with 1-3 substituents selected from halogen, cyano, amino, hydroxyl, carbonyl, and C1-6 alkyl; R3 is —NH—R5, and R5 is 3-7 membered heterocyclyl substituted with 1-2 substituents selected from C1-6 alkyl.
On the basis of the above technical solutions, for the compound, or the pharmaceutically acceptable salt, the stereoisomer or the tautomer thereof provided in the present invention, wherein R1 and R2, together with the carbon atom to which they are attached, form a 5-8 membered ring A, wherein the 5-8 membered ring A is selected from 5-8 membered cycloalkyl, 5-8 membered cycloalkenyl, 5-8 membered heterocyclyl, phenyl, and 5-8 membered heteroaryl; ring A is optionally substituted with 1-2 substituents selected from cyano, halogen, C1-6 alkyl, —NH—C1-6 alkyl, haloC1-6 alkyl, C1-6 alkoxy, 3-7 membered cycloalkyl, 5-7 membered heteroaryl, C1-6 alkylsulfonyl, and —N(C1-6 alkyl)2;
On the basis of the above technical solutions, for the compound, or the pharmaceutically acceptable salt, the stereoisomer or the tautomer thereof provided in the
ring A is optionally substituted with 1-2 substituents selected from cyano, halogen, C1-6 alkyl, —NH—C1-6 alkyl, haloC1-6 alkyl, C1-6 alkoxy, 3-7 membered cycloalkyl, 5-7 membered heteroaryl, C1-6 alkylsulfonyl, and —N(C1-6 alkyl)2.
On the basis of the above technical solutions, for the compound, or the pharmaceutically acceptable salt, the stereoisomer or the tautomer thereof provided in the present invention, wherein Y is selected from
Y is substituted with 1-3 substituents selected from halogen, cyano, hydroxyl, amino, C1-6 alkyl, C1-6 alkoxy, hydroxyl-substituted C1-6 alkyl, haloC1-6 alkyl, aminocarbonyl, C1-6 alkylamino, C1-6 alkylcarbonylamino, C1-6 alkylaminocarbonyl, 3-7 membered heterocyclyl, 3-7 membered cycloalkyl, 5-7 membered heteroaryl, C1-6 alkylsulfonyl, C1-6 alkylthio, and C1-6 alkylsulfinyl.
The present invention also provides the aforementioned compound of formula (I) or (II), the pharmaceutically acceptable salt thereof, or the stereoisomer or the tautomer thereof as shown in Table 1:
The technical solutions of the present invention also include a pharmaceutical composition comprising any of the above compounds, or the pharmaceutically acceptable salt, the stereoisomer or the tautomer thereof, and a pharmaceutically acceptable carrier.
The technical solutions of the present invention also include use of the compound of general formula (I) or general formula (II), or the pharmaceutically acceptable salt, the stereoisomer or the tautomer thereof, or the above pharmaceutical composition in the manufacture of a medicament for preventing and/or treating NLRP3 inflammasome-associated diseases.
The technical solutions of the present invention also include use of the above compound, or the pharmaceutically acceptable salt, the stereoisomer or the tautomer thereof, or the above pharmaceutical composition thereof in the manufacture of a medicament for preventing and/or treating inflammasome-associated diseases, immune diseases, inflammatory diseases, autoimmune diseases, or autoinflammatory diseases.
The “halogen” described herein refers to fluorine, chlorine, bromine, and iodine.
The “hydroxyl” described herein refers to —OH group.
The “cyano” described herein refers to —CN group.
The “amino” described herein refers to —NH2 group.
The “carboxyl” described herein refers to —COOH group.
The “nitro” described herein refers to —NO2 group.
The “C1-6 alkyl” described herein refers to linear or branched alkyl derived by removing one hydrogen atom from a hydrocarbon moiety containing 1 to 6 carbon atoms, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, 2-methylbutyl, neopentyl, 1-ethylpropyl, n-hexyl, isohexyl, 4-methylpentyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 3,3-dimethylbutyl, 2,2-dimethylbutyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 1,3-dimethylbutyl, 2,3-dimethylbutyl, 2-ethylbutyl, and 1-methyl-2-methylpropyl.
The “ene” of “C1-6 alkylene” described herein refers to a divalent group derived by removing two hydrogen atoms from a C1-6 alkyl group.
The “haloC1-6 alkyl” described herein refers to a C1-C6 alkyl group substituted with one or more halogen groups as defined above. Examples of haloC1-6 alkyl include, but are not limited to, trifluoromethyl, difluoromethyl, fluoromethyl, trichloromethyl, 2,2,2-trifluoroethyl, 1,3-dibromopropan-2-yl, 3-bromo-2-fluoropropyl, and 1,4,4-trifluorobutan-2-yl.
The “C1-6 alkoxy” described herein refers to a group in which the “C1-6 alkyl” defined above is linked to a parent molecule via an oxygen atom, i.e., a “C1-6 alkyl-O—” group, such as methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, tert-butoxy, n-pentyloxy, neopentyloxy and n-hexyloxy.
The “haloC1-6 alkoxy” described herein refers to a C1-C6 alkoxy group substituted with one or more halogen groups as defined above, examples of which include, but are not limited to, fluoromethoxy, chloromethoxy, trifluoromethoxy, trifluoroethoxy, fluoroethoxy, and fluoropropoxy.
The “C2-6 alkenyl” described herein refers to linear or branched alkenyl derived by removing one hydrogen atom from an alkene moiety containing 2-6 carbon atoms and at least one carbon-carbon double bond, such as vinyl, 1-propenyl, 2-propenyl, 1-butenyl, 2-butenyl, 1,3-butadien-1-yl, 1-penten-3-yl, 2-penten-1-yl, 3-penten-1-yl, 3-penten-2-yl, 1,3-pentadien-1-yl, 1,4-pentadien-3-yl, 1-hexen-3-yl, and 1,4-hexadien-1-yl. Preferably, “C2-6 alkenyl” contains one carbon-carbon double bond.
The “C2-6 alkynyl” described herein refers to linear or branched alkynyl derived by removing one hydrogen atom from an alkyne moiety containing 2-6 carbon atoms and at least one carbon-carbon triple bond, such as ethynyl, propynyl, butynyl, pentynyl, and hexynyl. Preferably, “C2-6 alkynyl” contains one carbon-carbon triple bond.
The “C1-6 alkylamino”, “C1-6 alkylcarbonylamino”, “C1-6 alkylsulfonyl” and “aminocarbonyl” described herein refer to C1-6 alkyl-NH— group, C1-6 alkyl-C(O)—NH— group, C1-6 alkyl-S(O)2— group, and NH2—C(O)— group, respectively.
The “5-12 membered ring” described herein includes carbon rings or heterocyclic rings which may be chemically formed, such as 5-12 membered cycloalkyl, 5-7 membered cycloalkyl, 5-12 membered cycloalkenyl, 5-7 membered cycloalkenyl, 6-12 membered fused cycloalkyl, 5-12 membered heterocyclyl, 5-7 membered heterocyclyl, 6-12 membered fused heterocycle, aryl, 5-12 membered heteroaryl, 8-12 membered fused heteroaryl, and 5-7 membered heteroaryl.
The “3-12 membered cycloalkyl” described herein refers to a monovalent group or (as required) divalent group (e.g, 5-12 membered cycloalkyl) derived from 3-12 membered cycloalkane, which may be a monocyclic, bicyclic or fused cycloalkyl system. Unless otherwise specified, a certain membered cycloalkyl group (e.g., 5-12 membered cycloalkyl, 5-8 membered cycloalkyl, 5-7 membered cycloalkyl, 3-7 membered cycloalkyl, 3-6 membered cycloalkyl, or 4-6 membered cycloalkyl) includes all monocyclic, fused rings (e.g., 6-12 membered fused cycloalkyl) which may be formed, including cases where they are fused in the ortho-fused, spiro- and bridged form.
A monocyclic system is typically a cyclic hydrocarbon group containing 3 to 12 carbon atoms (such as 3 to 8 or 3 to 6 carbon atoms). Examples of cycloalkyl include, but are not limited to: cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclopentyl-1,3-diyl, cyclohexyl-1,4-diyl, and cycloheptyl-1,4-diyl. Fused cycloalkyl includes ortho-fused cycloalkyl, bridged cycloalkyl and spiro-cycloalkyl. Ortho-fused cycloalkyl may be 6-11 membered ortho-fused cycloalkyl (such as 7-10 membered ortho-fused cycloalkyl), and the representative examples include, but are not limited to, bicyclo[3.1.1]heptane, bicyclo[2.2.1]heptane, bicyclo[2.2.2]octane, bicyclo[3.2.2]nonane, bicyclo[3.3.1]nonane and bicyclo[4.2.1]nonyl. Spiro-cycloalkyl may be 7-12 membered spiro-cycloalkyl (such as 7-11 membered spiro-cycloalkyl), examples of which include, but are not limited to
groups. Bridged cycloalkyl may be 6-10 membered bridged cycloalkyl (such as 7-10 membered bridged cycloalkyl), examples of which include, but are not limited to:
groups.
The “3-7 membered cycloalkyl” described herein refers to a monovalent group or (as required) divalent group derived from 3-7 membered cycloalkane. The “3-7 membered cycloalkyl” may be 3, 4, 5, 6, or 7 membered cycloalkyl, and examples of the 3-7 membered cycloalkyl include cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.
The “cycloalkenyl” described herein refers to a group obtained by having at least one double bond in the above cycloalkyl. It may be, for example, “3-12 membered cycloalkenyl”, i.e., may have 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 ring-forming carbon atoms. Unless otherwise specified, a certain membered cycloalkenyl encompasses all possible monocyclic or fused cycloalkenyls (including ortho-fused, spiro and bridged). Cycloalkenyl may be 3-12 membered cycloalkenyl, 3-8 membered cycloalkenyl, 5-8 membered cycloalkenyl, 5-7 membered cycloalkenyl, 4-6 membered cycloalkenyl, 7-11 membered spiro-cycloalkenyl, 7-11 membered ortho-fused cycloalkenyl, 6-11 membered bridged cycloalkenyl, etc. Examples of cycloalkenyl include, but are not limited to, cyclobutenyl, cyclopentenyl, cyclopentadienyl, cyclohexenyl, 1,4-cyclohexadien-1-yl, cycloheptenyl, 1,4-cycloheptadien-1-yl, cyclooctenyl and 1,5-cyclooctadien-1-yl.
The “5-7 membered cycloalkenyl” described herein refers to a group obtained by having at least one double bond in a 5-7 membered cycloalkyl group, such as cyclobutenyl, cyclopentenyl, cyclohexenyl, and cycloheptenyl.
The “3-14 membered heterocyclyl” described herein refers to a monovalent group or (as required) divalent group derived from 3-14 membered heterocycloalkane, i.e., a non-aromatic cyclic group obtained by substituting at least one ring carbon atom of 3-14 membered heterocycloalkane with a heteroatom selected from O, S, S(O), S(O)2, C(O) and N, which preferably contains 1-3 heteroatoms. The “3-14 membered heterocyclyl” (e.g., 5-14 membered heterocyclyl, or 5-12 membered heterocyclyl) includes monocyclic heterocyclyl, bicyclic heterocyclyl systems, or fused heterocyclic systems in which one or more rings may be saturated or partially saturated, but does not include aromatic rings. Unless otherwise specified, a certain membered heterocyclyl group (e.g., 3-8 membered heterocyclyl, 3-7 membered heterocyclyl, 5-8 membered heterocyclyl, 5-7 membered heterocyclyl, 5-6 membered heterocyclyl, 4-6 membered heterocyclyl, or 6 membered heterocyclyl) includes all monocyclic, fused ring (including fusion in the ortho-fused, spiro- and bridged form), and saturated and partially saturated cases which may be formed.
Monocyclic heterocyclyl may be 3-8 membered heterocyclyl (such as 5-7 membered heterocyclyl, 3-7 membered heterocyclyl, 4-7 membered heterocyclyl, or 5-6 membered heterocyclyl), 3-8 membered nitrogen-containing heterocyclyl (such as 4-7 membered nitrogen-containing heterocyclyl, or 5-6 membered nitrogen-containing heterocyclyl), or 3-8 membered saturated heterocyclyl (such as 5-6 membered saturated heterocyclyl), examples of which include, but are not limited to, azacyclopropyl, ozacyclopropyl, thiocyclopropyl, azacyclobutyl, oxacyclobutyl, thiocyclobutyl, tetrahydrofuranyl, tetrahydropyrrolyl, tetrahydrothienyl, imidazolidinyl, pyrazolidinyl, 1,2-oxazolidinyl, 1,3-oxazolidinyl, 1,2-thiazolidinyl, 1,3-thiazolidinyl, tetrahydro-2H-pyranyl, tetrahydro-2H-thiopyranyl, piperidinyl, piperazinyl, morpholinyl, 1,4-dioxanyl, 1,4-oxathianyl, 4,5-dihydroisoxazolyl, 4,5-dihydrooxazolyl, 2,5-dihydrooxazolyl, 2,3-dihydrooxazolyl, 3,4-dihydro-2H-pyrrolyl, 2,3-dihydro-1H-pyrrolyl, 2,5-dihydro-1H-imidazolyl, 4,5-dihydro-1H-imidazolyl, 4,5-dihydro-1H-pyrazolyl, 4,5-dihydro-3H-pyrazolyl, 4,5-dihydrothiazolyl, 2,5-dihydrothiazolyl, 2H-pyranyl, 4H-pyranyl, 2H-thiopyranyl, 4H-thiopyranyl, 2,3,4,5-tetrahydropyridyl, 1,2-isoxazolyl, 1,4-isoxazolyl, and 6H-1,3-oxazinyl.
Fused heterocyclyl (e.g., 6-12 membered fused heterocyclyl) includes ortho-fused heterocyclyl, spiro-heterocyclyl, and bridged heterocyclyl, which may be saturated, partially saturated or unsaturated, but non-aromatic. Fused heterocyclyl may be a 5-6 membered monocyclic heterocyclyl ring which is fused to a benzene ring, 5-6 membered monocyclic cycloalkyl, 5-6 membered monocyclic cycloalkenyl, 5-6 membered monocyclic heterocyclyl or 5-6 membered monocyclic heteroaryl.
The ortho-fused heterocyclyl may be 6-12 membered ortho-fused heterocyclyl (such as 6-11 membered ortho-fused heterocyclyl or 7-10 membered ortho-fused heterocyclyl), 6-11 membered saturated ortho-fused heterocyclyl or 6-11 membered nitrogen-containing ortho-fused heterocyclyl, and the representative examples thereof include, but are not limited to: 3-azabicyclo[3.1.0]hexyl, 3,6-diazabicyclo[3.2.0]heptyl, 3,8-diazabicyclo[4.2.0]octyl, 3,7-diazabicyclo[4.2.0]octyl, octahydropyrrolo[3,4-c]pyrrolyl, octahydropyrrolo[3,4-b]pyrrolyl, octahydropyrrolo[3,4-b][1,4]oxazinyl, octahydro-1H-pyrrolo[3,4-c]pyridyl, 2,3-dihydrobenzofuran-2-yl, 2,3-dihydrobenzofuran-3-yl, indolin-1-yl, indolin-2-yl, indolin-3-yl, 2,3-dihydrobenzothiophen-2-yl, octahydro-1H-indolyl, and octahydrobenzofuranyl.
The spiro-heterocyclyl may be 6-12 membered spiro-heterocyclyl (such as 7-12 membered spiro-heterocyclyl), 7-12 membered saturated spiro-heterocyclyl or 7-12 membered nitrogen-containing spiro-heterocyclyl, and the examples include, but are not limited to:
The bridged heterocyclyl may be 6-12 membered bridged heterocyclyl (such as 6-10 membered bridged heterocyclyl (e.g., 6-10 membered nitrogen-containing bridged heterocyclyl, particularly 7-membered nitrogen-containing bridged heterocyclyl), or 7-10 membered bridged heterocyclyl), examples of which include, but are not limited to:
The “aryl” described herein refers to a monovalent or (as required) divalent cyclic aromatic group containing 6-14 carbon atoms derived from aromatic carbocyclic hydrocarbon, including phenyl, naphthyl, phenanthryl, etc.
The “5-14 membered heteroaryl” described herein refers to an aromatic 5-14 membered cyclic group in which at least one ring carbon atom is substituted with a heteroatom selected from O, S and N, which may be 5, 6, 7, 8, 9, 10, 11, 12, 13 or 14 membered heteroaryl and preferably contains 1-3 heteroatoms. Moreover, the case where carbon atoms or sulfur atoms are oxidized or nitridized is included. For example, carbon atoms are substituted with C(O) and sulfur atoms are substituted with S(O) or S(O)2. Heteroaryl includes monocyclic heteroaryl and fused heteroaryl. Unless otherwise specified, a certain membered heteroaryl includes all possibly formed monocyclic, polycyclic, fully aromatic and partially aromatic cases. Monocyclic heteroaryl may be 5-7 membered heteroaryl (such as 5-6 membered heteroaryl), examples of which include, but are not limited to, furanyl, imidazolyl, isoxazolyl, thiazolyl, isothiazolyl, oxadiazolyl, oxazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, pyrazolyl, pyrrolyl, tetrazolyl, thiadiazolyl, thienyl, triazolyl and triazinyl.
In some embodiments, fused heteroaryl refers to a group which is formed by fusing a monocyclic heteroaromatic ring to phenyl, cycloalkenyl, heteroaryl, cycloalkyl or heterocyclyl. In some embodiments, fused heteroaryl (such as 8-14 membered fused heteroaryl) may be 8-14 membered ortho-fused heteroaryl (such as 9-10 membered ortho-fused heteroaryl), and the examples include, but are not limited to, benzimidazolyl, benzofuranyl, benzothienyl, benzoxadiazolyl, benzothiadiazolyl, benzothiazolyl, cinnolinyl, 5,6-dihydroquinolin-2-yl, 5,6-dihydroisoquinolin-1-yl, furopyridinyl, indazolyl, indolyl, isoindolyl, isoquinolinyl, naphthyridinyl, purinyl, quinolinyl, 5,6,7,8-tetrahydroquinolin-2-yl, 5,6,7,8-tetrahydroquinolinyl, 5,6,7,8-tetrahydroquinolin-4-yl, 5,6,7,8-tetrahydroisoquinolin-1-yl, thienopyridyl, 4,5,6,7-tetrahydro[c][1,2,5]oxadiazolyl and 6,7-dihydro[c][1,2,5]oxadiazol-4(5H)keto.
The “pharmaceutically acceptable salt” described herein refers to a pharmaceutically acceptable addition salt of acid and base and a solvate. Such pharmaceutically acceptable salts include salts of the following acids: hydrochloric acid, phosphoric acid, hydrobromic acid, sulfuric acid, sulfurous acid, formic acid, toluenesulfonic acid, methanesulfonic acid, nitric acid, benzoic acid, citric acid, tartaric acid, maleic acid, hydroiodic acid, alkanoic acid (such as acetic acid, HOOC—(CH2)n-COOH (wherein n is 0-4)), etc. Such pharmaceutically acceptable salts further include salts of the following bases: sodium, potassium, calcium, ammonium, etc. Those skilled in the art know a variety of pharmaceutically acceptable non-toxic addition salts.
The term “optionally” means that the subsequently described event may or may not occur, and that the description includes both the occurrence and non-occurrence of the event.
All numerical ranges described herein include both endpoints of the ranges, all integers within the ranges, and subranges formed by these integers. For example, “3-7 membered” includes 3, 4, 5, 6, and 7 membered; “0-4” includes 0, 1, 2, 3, and 4.
The “stereoisomer” of the compound described herein refers to an isomer resulting from the different spatial arrangement of the atoms in a molecule. An enantiomer will be produced when an asymmetric carbon atom is present in a compound, or a cis-trans isomer will be produced when a carbon-carbon double bond or a ring structure is present in a compound.
The term “tautomer” refers to a particular functional group isomer in which different functional group isomers are in dynamic equilibrium and are rapidly transformed into each other. For example, in the presence of a ketone or oxime, a tautomer will be produced, and representative examples are: keto-enol tautomers, phenol-keto tautomers, nitroso-oxime tautomers, imine-enamine tautomers, etc.
All enantiomers, diastereomers, racemates, cis-trans isomers, tautomers, geometric isomers, and epimers of all of the compounds, and mixtures thereof are included in the scope of the present invention.
In the chemical configuration of the compound of the present invention, the bond “/” represents an unspecified configuration, namely, if a chiral isomer is present in a chemical structure, the bond “” may be “
” or “
”, or includes both “
” and “
” In the chemical structure of the compound of the present invention, “
” indicates the point linked to the parent molecule.
The present invention provides a compound of general formula (I), or a pharmaceutically acceptable salt, a stereoisomer or a tautomer thereof:
In some embodiments, R1 and R2, together with the carbon atom to which they are attached, form a 5-12 membered ring A, wherein the 5-12 membered ring A is optionally substituted with —N(C1-6 alkyl)2.
In some embodiments, R3 is —(C1-6 alkylene)0-2-O—R5.
In some embodiments, R5 is optionally substituted with a substituent selected from C2-6 alkenylcarbonyl, sulfonyl, and C1-6 alkylcarbonyl.
In some embodiments, R5 is optionally substituted with carboxyl.
In some embodiments, Y is optionally substituted with a substituent selected from C1-6 alkylaminocarbonyl, and sulfonyl.
In some embodiments, Y is optionally substituted with a substituent selected from C1-6 alkylthio and C1-6 alkylsulfinyl.
In some embodiments, when R5 is substituted, the substituents on R5, which are C1-6 alkyl, haloC1-6 alkyl, C1-6 alkoxy, 3-7 membered heterocyclyl, 3-7 membered cycloalkyl, aryl, 5-7 membered heteroaryl, and sulfonyl, are optionally substituted with 1-3 substituents selected from halogen, cyano, amino, hydroxyl, carbonyl, C1-6 alkyl, and 3-6 membered cycloalkyl;
In some embodiments, R1 and R2, together with the carbon atom to which they are attached, form a 5-12 membered ring A, wherein the 5-12 membered ring A is optionally substituted with —N(C1-6 alkyl)2; R5 is optionally substituted with a substituent selected from C2-6 alkenylcarbonyl, sulfonyl, and C1-6 alkylcarbonyl; Y is optionally substituted with a substituent selected from C1-6 alkylaminocarbonyl and sulfonyl; the substituents on R5 and Y, which are C1-6 alkyl, haloC1-6 alkyl, C1-6 alkoxy, 3-7 membered heterocyclyl, 3-7 membered cycloalkyl, aryl, 5-7 membered heteroaryl, and sulfonyl, are optionally substituted with 1-3 substituents selected from halogen, cyano, amino, hydroxyl, carbonyl, C1-6 alkyl, and 3-6 membered cycloalkyl.
In some embodiments, ring A is selected from 5-12 membered cycloalkyl, 5-12 membered cycloalkenyl, 5-12 membered heterocyclyl, aryl, and 5-12 membered heteroaryl.
In some embodiments, ring A is selected from 5-8 membered cycloalkyl, 5-8 membered cycloalkenyl, 5-8 membered heterocyclyl, phenyl, and 5-8 membered heteroaryl.
In some embodiments, ring A is selected from phenyl, and 5-6 membered heteroaryl, 5-8 membered cycloalkyl, 5-8 membered cycloalkenyl and 5-8 membered heterocyclyl containing 1-2 heteroatoms selected from O, S, and N.
In some embodiments, ring A is optionally substituted with 1-4 substituents selected from hydroxyl, amino, carboxyl, cyano, nitro, halogen, carbonyl, C1-6 alkyl, —NH—C1-6 alkyl, haloC1-6 alkyl, C1-6 alkoxy, haloC1-6 alkoxy, 3-7 membered heterocyclyl, 3-7 membered cycloalkyl, aryl, 5-7 membered heteroaryl, C1-6 alkylsulfonyl, and —N(C1-6 alkyl)2.
In some embodiments, ring A is unsubstituted.
In some embodiments, ring A is substituted with 1-2 substituents selected from cyano, halogen, C1-6 alkyl, —NH—C1-6 alkyl, haloC1-6 alkyl, C1-6 alkoxy, 3-7 membered cycloalkyl, 5-7 membered heteroaryl, C1-6 alkylsulfonyl, and —N(C1-6 alkyl)2;
In some embodiments, ring A is selected from 5-8 membered cycloalkyl, 5-8 membered cycloalkenyl, 5-8 membered heterocyclyl, phenyl, and 5-6 membered heteroaryl; ring A is optionally substituted with 1-2 substituents selected from cyano, halogen, C1-6 alkyl, —NH—C1-6 alkyl, haloC1-6 alkyl, C1-6 alkoxy, 3-7 membered cycloalkyl, 5-7 membered heteroaryl, C1-6 alkylsulfonyl, and —N(C1-6 alkyl)2.
In some embodiments, the present invention relates to the aforementioned compound, or the pharmaceutically acceptable salt, the stereoisomer or the tautomer thereof, which has the structure of general formula (II):
In some embodiments, the present invention relates to the aforementioned compound, or the pharmaceutically acceptable salt, the stereoisomer or the tautomer thereof, wherein ring A is selected from phenyl and 5-7 membered heteroaryl; ring A is optionally substituted with 1-4 substituents selected from hydroxyl, amino, carboxyl, cyano, nitro, halogen, C1-6 alkyl, —NH—C1-6 alkyl, haloC1-6 alkyl, C1-6 alkoxy, haloC1-6 alkoxy, 3-7 membered heterocyclyl, 3-7 membered cycloalkyl, and C1-6 alkylsulfonyl. Furthermore, ring A is optionally substituted with —N(C1-6 alkyl)2.
In some embodiments, the present invention relates to the aforementioned compound, or the pharmaceutically acceptable salt, the stereoisomer or the tautomer thereof, wherein ring A is selected from
ring A is optionally substituted with 1-4 substituents selected from hydroxyl, amino, carboxyl, cyano, nitro, halogen, C1-6 alkyl, —NH—C1-6 alkyl, haloC1-6 alkyl, C1-6 alkoxy, haloC1-6 alkoxy, 3-7 membered heterocyclyl, 3-7 membered cycloalkyl, C1-6 alkylsulfonyl, and —N(C1-6 alkyl)2.
In some embodiments, ring A is selected from
ring A is optionally substituted with 1-2 substituents selected from cyano, halogen, C1-6 alkyl, —NH—C1-6 alkyl, haloC1-6 alkyl, C1-6 alkoxy, 3-7 membered cycloalkyl, 5-7 membered heteroaryl, C1-6 alkylsulfonyl, and —N(C1-6 alkyl)2.
In some embodiments, ring A is
X1 is selected from H, CH3, F, Cl, Br, —OCH3, CN, —CF3, —NHCH3, and
X2 is selected from H, CH3, F, Cl, Br, —OCH3, CN, —CF3, —NHCH3,
preferably, ring A is further unsubstituted.
In some embodiments, Y is selected from aryl, 5-14 membered heteroaryl, 3-14 membered heterocyclyl, and 3-12 membered cycloalkyl, and Y is optionally substituted with 1-3 substituents selected from halogen, cyano, amino, hydroxyl, carbonyl, C1-6 alkyl, haloC1-6 alkyl, C1-6 alkoxy, 3-7 membered heterocyclyl, 3-7 membered cycloalkyl, aryl, 5-7 membered heteroaryl, C1-6 alkylamino, C1-6 alkylcarbonylamino, C1-6 alkylsulfonyl, aminocarbonyl, C1-6 alkylaminocarbonyl, sulfonyl, C1-6 alkylthio, and C1-6 alkylsulfinyl.
In some embodiments, the present invention relates to the aforementioned compound, or the pharmaceutically acceptable salt, the stereoisomer or the tautomer thereof,
In some embodiments, Y is optionally substituted with a substituent of C1-6 alkylaminocarbonyl or sulfonyl.
In some embodiments, when Y is substituted, C1-6 alkyl, haloC1-6 alkyl, C1-6 alkoxy, 3-7 membered heterocyclyl, 3-7 membered cycloalkyl or sulfonyl on Y is optionally substituted with 1-3 substituents selected from halogen, cyano, amino, hydroxyl, carbonyl, and C1-6 alkyl.
In some embodiments, Y is selected from phenyl, 5-8 membered heteroaryl, 3-8 membered heterocyclyl, and 3-7 membered cycloalkyl.
In some embodiments, Y is selected from phenyl and 5-7 membered heteroaryl.
In some embodiments, Y is selected from phenyl, and 5-6 membered heteroaryl containing 1-2 N heteroatoms.
In some embodiments, Y is optionally substituted with 1-3 substituents selected from halogen, cyano, amino, hydroxyl, carbonyl, C1-6 alkyl, haloC1-6 alkyl, C1-6 alkoxy, 3-7 membered heterocyclyl, 3-7 membered cycloalkyl, aryl, 5-7 membered heteroaryl, C1-6 alkylamino, C1-6 alkylcarbonylamino, C1-6 alkylsulfonyl, aminocarbonyl, C1-6 alkylaminocarbonyl, sulfonyl, C1-6 alkylthio, and C1-6 alkylsulfinyl.
In some embodiments, when Y is substituted, the substituents on Y, which are C1-6 alkyl, haloC1-6 alkyl, C1-6 alkoxy, 3-7 membered heterocyclyl, 3-7 membered cycloalkyl, aryl, 5-7 membered heteroaryl, and sulfonyl, are optionally substituted with 1-3 substituents selected from halogen, cyano, amino, hydroxyl, carbonyl, C1-6 alkyl, and 3-6 membered cycloalkyl.
In some embodiments, Y is substituted with 1-3 substituents selected from halogen, cyano, hydroxyl, amino, C1-6 alkyl, C1-6 alkoxy, hydroxyl-substituted C1-6 alkyl, haloC1-6 alkyl, aminocarbonyl, C1-6 alkylamino, C1-6 alkylcarbonylamino, C1-6 alkylaminocarbonyl, 3-7 membered heterocyclyl, 3-7 membered cycloalkyl, 5-7 membered heteroaryl, C1-6 alkylsulfonyl, C1-6 alkylthio, and C1-6 alkylsulfinyl.
In some embodiments, Y is substituted with cyano and is optionally substituted with 1-2 substituents selected from halogen, cyano, amino, hydroxyl, carbonyl, C1-6 alkyl, haloC1-6 alkyl, C1-6 alkoxy, 3-7 membered heterocyclyl, 3-7 membered cycloalkyl, C1-6 alkylamino, C1-6 alkylcarbonylamino, C1-6 alkylsulfonyl, aminocarbonyl, C1-6 alkylaminocarbonyl, and sulfonyl; the C1-6 alkyl, haloC1-6 alkyl, C1-6 alkoxy, 3-7-membered heterocyclyl, 3-7-membered cycloalkyl and sulfonyl are optionally substituted with 1-3 substituents selected from halogen, cyano, amino, hydroxyl, carbonyl, and C1-6 alkyl.
In some embodiments, Y is substituted with 1 cyano and is optionally substituted with 1-2 substituents selected from halogen, hydroxyl, amino, C1-6 alkyl, C1-6 alkoxy, hydroxyl-substituted C1-6 alkyl, haloC1-6 alkyl, aminocarbonyl, C1-6 alkylamino, C1-6 alkylcarbonylamino, C1-6 alkylaminocarbonyl, 3-7 membered heterocyclyl, 3-7 membered cycloalkyl, 5-7 membered heteroaryl, C1-6 alkylsulfonyl, C1-6 alkylthio, and C1-6 alkylsulfinyl.
One embodiment of the present invention relates to the aforementioned compound, or the pharmaceutically acceptable salt, the stereoisomer or the tautomer thereof, wherein Y is selected from
Y is substituted with 1-3 substituents selected from halogen, cyano, hydroxyl, amino, C1-6 alkyl, C1-6 alkoxy, hydroxyl-substituted C1-6 alkyl, haloC1-6 alkyl, aminocarbonyl, C1-6 alkylamino, C1-6 alkylcarbonylamino, C1-6 alkylaminocarbonyl, 3-7 membered heterocyclyl, 3-7 membered cycloalkyl, 5-7 membered heteroaryl, C1-6 alkylsulfonyl, C1-6 alkylthio, and C1-6 alkylsulfinyl.
In some embodiments, the present invention relates to the aforementioned compound, or the pharmaceutically acceptable salt, the stereoisomer or the tautomer thereof,
In some embodiments, R3 is selected from —(C1-6 alkylene)0-2-NR4R5, —(C1-6 alkylene)0-2-NR4—COR5, —(C1-6 alkylene)0-2-CO—NR4-R5, and —(C1-6 alkylene)0-2-O—R5; R4 is selected from hydrogen and C1-6 alkyl; R5 is selected from 3-7 membered heterocyclyl, 3-7 membered cycloalkyl, aryl, and 5-7 membered heteroaryl.
In some embodiments, R4 is hydrogen.
In some embodiments, R4 is C1-6 alkyl.
In some embodiments, R4 is selected from methyl and ethyl.
In some embodiments, R3 is selected from —NR4R5, —NH—COR5, and —O—R5; R4 is selected from hydrogen and C1-3 alkyl; R5 is selected from 3-7 membered cycloalkyl and 3-7 membered heterocyclyl.
In some embodiments, R3 is —NHR5, and R5 is selected from 4-6 membered cycloalkyl and 4-6 membered heterocyclyl.
In some embodiments, R5 is selected from 4-6 membered cycloalkyl, and 4-6 membered heterocyclyl containing 1 heteroatom selected from O, N, and S.
In some embodiments, R5 is optionally substituted with 1-4 substituents selected from halogen, cyano, amino, hydroxyl, C1-6 alkyl, haloC1-6 alkyl, C1-6 alkoxy, 3-7 membered heterocyclyl, 3-7 membered cycloalkyl, aryl, 5-7 membered heteroaryl, C2-6 alkenylcarbonyl, sulfonyl, C1-6 alkylcarbonyl, and carboxyl; when R5 is substituted, the substituents on R5, which are C1-6 alkyl, haloC1-6 alkyl, C1-6 alkoxy, 3-7 membered heterocyclyl, 3-7 membered cycloalkyl, aryl, 5-7 membered heteroaryl, and sulfonyl, are optionally substituted with 1-3 substituents selected from halogen, cyano, amino, hydroxyl, carbonyl, C1-6 alkyl, and 3-6 membered cycloalkyl.
In some embodiments, R5 is optionally substituted with 1-2 substituents selected from C1-6 alkyl, hydroxyl, hydroxyl-substituted C1-6 alkyl, 3-7 membered cycloalkyl-substituted C1-6 alkyl, hydroxyl, halogen, C2-6 alkenylcarbonyl, C1-6 alkylsulfonyl, 3-7 membered cycloalkylsulfonyl, aminosulfonyl, 3-7 membered heterocyclyl, 3-7 membered cycloalkyl, carboxyl, and C1-6 alkylcarbonyl.
In some embodiments, the present invention relates to the aforementioned compound, or the pharmaceutically acceptable salt, the stereoisomer or the tautomer thereof, wherein R3 is selected from —NR4R5, —NH—COR5, and —O—R5; R4 is selected from hydrogen and C1-3 alkyl; R5 is selected from 3-7 membered cycloalkyl and 3-7 membered heterocyclyl, and R5 is optionally substituted with 1-2 substituents selected from C1-6 alkyl, hydroxyl, hydroxyl-substituted C1-6 alkyl, 3-7 membered cycloalkyl-substituted C1-6 alkyl, hydroxyl, halogen, C2-6 alkenylcarbonyl, C1-6 alkylsulfonyl, 3-7 membered cycloalkylsulfonyl, aminosulfonyl, 3-7 membered heterocyclyl, 3-7 membered cycloalkyl, carboxyl, and C1-6 alkylcarbonyl.
In some embodiments, R5 is selected from
preferably, R5 is further unsubstituted.
preferably, R5 is further unsubstituted.
In some embodiments, the present invention relates to the aforementioned compound, or the pharmaceutically acceptable salt, the stereoisomer or the tautomer thereof, wherein R3 is —NH—R5, and R5 is 3-7 membered heterocyclyl substituted with 1-2 substituents selected from C1-6 alkyl; R5 is preferably 5-6 membered heterocyclyl substituted with 1-2 substituents selected from C1-6 alkyl.
In some embodiments, the present invention relates to the aforementioned compound, or the pharmaceutically acceptable salt, the stereoisomer or the tautomer thereof, wherein ring A is selected from phenyl and 5-7 membered heteroaryl; ring A is optionally substituted with 1-4 substituents selected from hydroxyl, amino, carboxyl, cyano, nitro, halogen, C1-6 alkyl, —NH—C1-6 alkyl, haloC1-6 alkyl, C1-6 alkoxy, haloC1-6 alkoxy, 3-7 membered heterocyclyl, and 3-7 membered cycloalkyl;
In some embodiments, Y is optionally substituted with a substituent of C1-6 alkylaminocarbonyl or sulfonyl.
In some embodiments, when R5 is substituted, the substituents on R5, which are C1-6 alkyl, haloC1-6 alkyl, C1-6 alkoxy, 3-7 membered heterocyclyl, 3-7 membered cycloalkyl, and sulfonyl, are optionally substituted with 1-3 substituents selected from halogen, cyano, amino, hydroxyl, carbonyl, and C1-6 alkyl.
In some embodiments, the present invention relates to the aforementioned compound, or the pharmaceutically acceptable salt, the stereoisomer or the tautomer thereof, wherein R1 and R2, together with the carbon atom to which they are attached, form a 5-8 membered ring A, wherein the 5-8 membered ring A is selected from 5-8 membered cycloalkyl, 5-8 membered cycloalkenyl, 5-8 membered heterocyclyl, phenyl, and 5-6 membered heteroaryl; ring A is optionally substituted with 1-2 substituents selected from cyano, halogen, C1-6 alkyl, —NH—C1-6 alkyl, haloC1-6 alkyl, C1-6 alkoxy, 3-7 membered cycloalkyl, 5-7 membered heteroaryl, C1-6 alkylsulfonyl, and —N(C1-6 alkyl)2;
On the basis of any of the aforementioned embodiments or technical solutions, when Y is substituted with chloro, R5 is 6 membered heterocyclyl.
On the basis of any of the aforementioned embodiments or technical solutions, Y is substituted with 1-3 substituents selected from halogen, cyano, amino, hydroxyl, carbonyl, C1-6 alkyl, haloC1-6 alkyl, C1-6 alkoxy, 3-7 membered heterocyclyl, 3-7 membered cycloalkyl, aryl, 5-7 membered heteroaryl, C1-6 alkylamino, C1-6 alkylcarbonylamino, C1-6 alkylsulfonyl, aminocarbonyl, C1-6 alkylaminocarbonyl, sulfonyl, C1-6 alkylthio, and C1-6 alkylsulfinyl; when Y is substituted, the substituents on Y, which are C1-6 alkyl, haloC1-6 alkyl, C1-6 alkoxy, 3-7 membered heterocyclyl, 3-7 membered cycloalkyl, aryl, 5-7 membered heteroaryl, and sulfonyl, are optionally substituted with 1-3 substituents selected from halogen, cyano, amino, hydroxyl, carbonyl, C1-6 alkyl, and 3-6 membered cycloalkyl.
On the basis of any of the aforementioned embodiments or technical solutions,
In some embodiments, the pharmaceutical composition may comprise one or more pharmaceutically acceptable carriers, and may be administered to a patient or subject in need of such treatment by oral, parenteral, rectal, or transpulmonary administration, and the like. For oral administration, the pharmaceutical composition can be prepared into a conventional solid formulation, such as tablets, capsules, pills and granules; or can be prepared into an oral liquid formulation, such as oral solutions, oral suspensions and syrups. In the preparation of an oral formulation, an appropriate filler, binder, disintegrant, lubricant, and the like may be added. For parenteral administration, the pharmaceutical composition can be formulated into an injection, including a solution injection, a sterile powder for injection, and a concentrated solution for injection. The injection can be produced by a conventional method existing in the pharmaceutical field, and during the preparation process, no additive may be added, or an appropriate additive may be added according to the properties of the medicament. For rectal administration, the pharmaceutical composition can be formulated into a suppository and the like. For transpulmonary administration, the pharmaceutical composition can be formulated into an inhalant, spray, or the like.
The compound disclosed herein can be prepared by a variety of methods including standard chemical methods. Unless otherwise stated, any variable defined above will continue to have the meaning defined above. Exemplary general synthesis methods are elaborated in the following schemes, and can be easily modified to prepare other compounds disclosed herein. In the following detailed description, methods for synthesizing specific compounds according to the present invention are described.
The compound of formula (II) may be prepared by coupling a compound of formula (a) with a compound of formula (III);
Provided below is a method for preparing a compound of formula (II) by reacting a compound of formula (a) with a compound of formula (III):
For example, the compound of formula (a) and the compound of formula (III) are added to a suitable solvent (such as 1,4-dioxane and water), a suitable catalyst (such as [1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium(II)) and a suitable base (such as sodium bicarbonate) are added, and the resulting mixture is heated and stirred at a suitable temperature (such as 90° C. to 110° C.) for a suitable period of time (such as 1 to 20 hours) under the protection of an inert gas (such as nitrogen). After the completion of reaction, the reaction solution is added to an appropriate amount of water, followed by extraction with a suitable extractant (such as ethyl acetate). After concentration under reduced pressure, the compound of formula (II) is separated out by using a suitable purification method (such as silica gel column chromatography or preparative thin-layer chromatography).
The compound of formula (a) may be prepared by coupling a compound of formula (b) with a compound of formula (IV);
The method for preparing the compound of formula (a) by reacting the compound of formula (b) with the compound of formula (IV) is as follows:
For example, the compound of formula (b) and the compound of formula (IV) are added to a suitable solvent (e.g., 1,4-dioxane), a suitable catalyst (e.g., tris(dibenzylideneacetone)dipalladium(0)), a suitable ligand (e.g., 1,1′-binaphthyl-2,2′-bisdiphenylphosphine), and a suitable base (e.g., cesium carbonate) are added, and the resulting mixture is heated and stirred at a suitable temperature (e.g., 90° C. to 110° C.) for a suitable period of time (e.g., 1 to 12 hours). After the completion of reaction, the reaction solution is poured into an appropriate amount of water, followed by extraction with a suitable extractant (such as ethyl acetate). After concentration under reduced pressure, the compound of formula (a) is separated out by using a suitable purification method (such as silica gel column chromatography or preparative thin-layer chromatography).
In some embodiments, the compound of formula (a) may be prepared by reacting the compound of formula (IV) with the compound of formula (b) under a heating condition.
For example, the compound of formula (b) and the compound of formula (IV) are added to a suitable solvent (e.g., N,N-dimethylacetamide), and the resulting mixture was stirred at a suitable temperature (e.g., 120° C.) for a suitable period of time (e.g., 16 to 20 hours). After the completion of reaction, the reaction solution is poured into an appropriate amount of water, followed by extraction with a suitable extractant (such as ethyl acetate). After concentration under reduced pressure, the compound of formula (a) is separated out by using a suitable purification method (such as silica gel column chromatography or preparative thin-layer chromatography).
In some embodiments, the compound of formula (II) may be prepared by coupling the compound of formula (i) with the compound of formula (IV);
The method for preparing the compound of formula (II) by reacting the compound of formula (i) with the compound of formula (IV) is as follows:
For example, the compound of formula (i) and the compound of formula (IV) are added to a suitable solvent (e.g., 1,4-dioxane), a suitable catalyst (e.g., tris(dibenzylideneacetone)dipalladium(0)), a suitable ligand (e.g., 1,1′-binaphthyl-2,2′-bisdiphenylphosphine), and a suitable base (e.g., cesium carbonate) are added, and the resulting mixture is heated and stirred at a suitable temperature (e.g., 90° C. to 110° C.) for a suitable period of time (e.g., 1 to 12 hours). After the completion of reaction, the reaction solution is poured into an appropriate amount of water, followed by extraction with a suitable extractant (such as ethyl acetate). After concentration under reduced pressure, the compound of formula (II) is separated out by using a suitable purification method (such as silica gel column chromatography or preparative thin-layer chromatography).
In some embodiments, the compound of formula (II) may be prepared by reacting the compound of formula (i) with the compound of formula (IV) under a heating condition.
For example, the compound of formula (i) and the compound of formula (IV) are added to a suitable solvent (e.g., N,N-dimethylacetamide), and the resulting mixture was stirred at a suitable temperature (e.g., 120° C.) for a suitable period of time (e.g., 16 to 20 hours). After the completion of reaction, the reaction solution is poured into an appropriate amount of water, followed by extraction with a suitable extractant (such as ethyl acetate). After concentration under reduced pressure, the compound of formula (II) is separated out by using a suitable purification method (such as silica gel column chromatography or preparative thin-layer chromatography).
The compound of formula (i) may be prepared by coupling a compound of formula (b) with a compound of formula (III);
The method for preparing the compound of formula (i) by reacting the compound of formula (b) with the compound of formula (III) is as follows:
For example, the compound of formula (b) and the compound of formula (III) are added to a suitable solvent (such as 1,4-dioxane and water), a suitable catalyst (such as [1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium(II)) and a suitable base (such as sodium bicarbonate) are added, and the resulting mixture is heated and stirred at a suitable temperature (such as 90° C. to 110° C.) for a suitable period of time (such as 1 to 20 hours) under the protection of an inert gas (such as nitrogen). After the completion of reaction, the reaction solution is added to an appropriate amount of water, followed by extraction with a suitable extractant (such as ethyl acetate). After concentration under reduced pressure, the compound of formula (i) is separated out by using a suitable purification method (such as silica gel column chromatography or preparative thin-layer chromatography).
In some embodiments, the compound of formula (b) may be prepared by halogenating a compound of formula (c);
The method for preparing the compound of formula (b) from the compound of formula (c) is as follows:
For example, the compound of formula (c) is added to a suitable solvent (e.g., acetonitrile), a halogenating agent (e.g., phosphorus oxychloride) is added, and the resulting mixture is heated and stirred at a suitable temperature (e.g., 25° C. to 90° C.) for a suitable period of time (e.g., 1 to 12 hours). After the completion of reaction, the reaction solution is poured into an appropriate amount of water, followed by extraction with a suitable extractant (such as ethyl acetate). After concentration under reduced pressure, the compound of formula (b) is separated out by using a suitable purification method (such as silica gel column chromatography or preparative thin-layer chromatography).
In some embodiments, the compound of formula (c) may be prepared from a compound of formula (d);
The method for preparing the compound of formula (c) from the compound of formula (d) is as follows:
For example, the compound of formula (d) (or an anhydride compound of ring A) is added to a suitable solvent (e.g., ethanol), hydrazine hydrate is added, and the resulting mixture is heated and stirred at a suitable temperature (e.g., 25° C. to 80° C.) for a suitable period of time (e.g., 0.5 to 5 hours). After the completion of reaction, the reaction solution is poured into an appropriate amount of water, followed by extraction with a suitable extractant (such as ethyl acetate). After concentration under reduced pressure, the compound of formula (c) is separated out by using a suitable purification method (such as silica gel column chromatography or preparative thin-layer chromatography).
In some embodiments, the compound of formula (i) may be prepared by the following reaction formula;
The specific method for preparing the compound of formula (i) is as follows:
For example, a compound of formula (e) is added to a suitable solvent (e.g., tetrahydrofuran), the resulting mixture is stirred at a suitable temperature (e.g., −70° C. to 0° C.) for a suitable period of time (e.g., 0.5 to 1 hour), and then a solution of a compound of formula (f) (e.g., tetrahydrofuran) is added at a suitable temperature (e.g., −70° C. to 0° C.) for a suitable period of time (e.g., 2 to 5 hours). After the completion of reaction, the reaction solution is poured into an appropriate amount of water, followed by extraction with a suitable extractant (such as ethyl acetate). After concentration under reduced pressure, the compound of formula (g) is separated out by using a suitable purification method (such as silica gel column chromatography or preparative thin-layer chromatography).
The compound of formula (g) is added to a suitable solvent (such as ethanol), hydrazine hydrate is added, and the resulting mixture is heated and stirred at a suitable temperature (such as 25° C. to 80° C.) for a suitable period of time (such as 0.5 to 5 hours). After the completion of reaction, the reaction solution is poured into an appropriate amount of water, followed by extraction with a suitable extractant (such as ethyl acetate). After concentration under reduced pressure, the compound of formula (h) is separated out by using a suitable purification method (such as silica gel column chromatography or preparative thin-layer chromatography).
The compound of formula (h) is added to a suitable solvent (e.g., acetonitrile), a halogenating agent (e.g., phosphorus oxychloride) is added, and the resulting mixture is heated and stirred at a suitable temperature (e.g., 25° C. to 90° C.) for a suitable period of time (e.g., 1 to 12 hours). After the completion of reaction, the reaction solution is poured into an appropriate amount of water, followed by extraction with a suitable extractant (such as ethyl acetate). After concentration under reduced pressure, the compound of formula (i) is separated out by using a suitable purification method (such as silica gel column chromatography or preparative thin-layer chromatography).
In some embodiments, the compound of formula (h) may be prepared by the following reaction formula;
For example, the compound of formula (m) is added to a suitable solvent (e.g., tetrahydrofuran), an organometallic reagent (e.g., isopropyl magnesium chloride) is added at a suitable temperature (e.g., −60° C. to 0° C.), and stirred for a suitable period of time (e.g., 0.5 to 2 hours), and then the solution is added to a solution of a compound of formula (j) (e.g., tetrahydrofuran) at a suitable temperature (e.g., −70° C. to 0° C.) and stirred for a suitable period of time (e.g., 2 to 5 hours). After the completion of reaction, the reaction solution is poured into an appropriate amount of water to adjust the pH, followed by extraction with a suitable extractant (such as ethyl acetate). After concentration under reduced pressure, a compound of formula (n) is separated out by using a suitable purification method (such as silica gel column chromatography or preparative thin-layer chromatography).
The compound of formula (n) is added to a suitable solvent (such as ethanol), hydrazine hydrate is added, and the resulting mixture is heated and stirred at a suitable temperature (such as 25° C. to 80° C.) for a suitable period of time (such as 0.5 to 5 hours). After the completion of reaction, the reaction solution is poured into an appropriate amount of water, followed by extraction with a suitable extractant (such as ethyl acetate). After concentration under reduced pressure, the compound of formula (h) is separated out by using a suitable purification method (such as silica gel column chromatography or preparative thin-layer chromatography).
The above preparation methods and the order of the respective steps in the methods are merely exemplary, and those skilled in the art can prepare the compounds of the present invention with reference to the above methods and can adjust the order of the steps as the case may be. In addition, the compounds whose sources are not described in the above preparation methods are generally commercially available, or may be synthesized from commercially available compounds by a method known to those skilled in the art.
The research finds that the compound provided in the present invention, or the pharmaceutically acceptable salt, the stereoisomer or the tautomer thereof has good inhibitory activity against the NLRP3 inflammasome, so that the compound of the present invention can be used for preventing and/or treating NLRP3 inflammasome-associated diseases.
In order to make the objective, technical schemes, and advantages of the present invention more apparent, the present invention is further described in detail below. It should be apparent that the examples described herein are only some examples of the present invention, but not all examples. Based on the examples of the present invention, all other examples obtained by those of ordinary skill in the art without creative work shall fall within the protection scope of the present invention.
The abbreviations and English expressions used in the present invention have the following meanings:
“THF” refers to tetrahydrofuran; “DMF” refers to N,N-dimethylformamide; “MeOH” refers to methanol; “EA” refers to ethyl acetate; “DCM” refers to dichloromethane; “DMA” refers to N,N-dimethylacetamide; “MTBE” refers to methyl tert-butyl ether; “EtOH” refers to ethanol; “DMAC” refers to dimethylacetamide; “PE” refers to petroleum ether; “n-BuLi” refers to n-butyllithium;
“FBS” refers to fetal bovine serum; “PBS” refers to phosphate-buffered saline solution; “PMA” refers to phorbol ester; “LPS” refers to lipopolysaccharide; “Nigericin” refers to nigericin sodium salt.
1-Bromo-2-methoxy-4-methylbenzene (8.0 g, 39.78 mmol, 1.0 eq.) was added to THE (60.0 mL), and the mixture was cooled to −70° C., followed by dropwise and slow addition of a solution of 2.5 M n-butyllithium in n-hexane (23.8 mL, 59.67 mmol, 1.5 eq.). The reaction solution was stirred at −70° C. for 2 h, triisopropyl borate (9.72 g, 51.72 mmol, 1.3 eq.) was added dropwise and slowly, and the resulting mixture was gradually warmed to room temperature and stirred for 4 h. After the reaction was completed as monitored by TLC, a 2 mol/L aqueous HCl solution (100.0 mL) was added to the system with stirring for 1 h, and EA (200.0 mL) was added, followed by liquid separation. The organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and slurried with petroleum ether (50.0 mL) for 1 h, and the residue was filtered under vacuum to give the product (5.8 g, yield: 87.8%).
(2-Methoxy-4-methylphenyl)boronic acid (5.19 g, 31.26 mmol, 1.0 eq.) was added to 1,4-dioxane (60.0 mL), 1,4-dichlorophthalazine (6.84 g, 34.39 mmol, 1.1 eq.) was added, followed by the addition of sodium bicarbonate (5.25 g, 62.52 mmol, 2.0 eq.), water (15.0 mL) and Pd(dppf)Cl2 (1.14 g, 1.56 mmol, 0.05 eq.), and the mixture was reacted at 90° C. for 5 h in nitrogen atmosphere. After the reaction was completed as monitored by TLC, EA (200.0 mL) and water (200 mL) were added to the system, followed by liquid separation. The organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (100-200 mesh silica gel, petroleum ether:ethyl acetate=10:1 to 4:1) to give the product (4.4 g, yield: 49.4%).
Step 3: Synthesis of 2-(4-bromophthalazin-1-yl)-5-methylphenol
1-Chloro-4-(2-methoxy-4-methylphenyl)phthalazine (4.0 g, 14.04 mmol, 1.0 eq.) was added to DCM (30.0 mL), the mixture was cooled to −65° C., followed by dropwise and slow addition of boron tribromide (7.0 g, 28.09 mmol, 2.0 eq.), and the mixture was gradually warmed to room temperature and stirred overnight. After the reaction was completed as monitored by TLC, methanol was added to the system at 0° C. to quench the reaction, and after concentration under reduced pressure, DCM and water were added for liquid separation. The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to give the product (2.3 g, yield: 52.2%).
2-(4-Bromophthalazin-1-yl)-5-methylphenol (2.3 g, 7.29 mmol, 1.0 eq.) was added to DMA (10.0 mL) and tert-butyl (R)-3-aminopiperidine-1-carboxylate (2.19 g, 10.94 mmol, 1.5 eq.), and the mixture was stirred at 120° C. overnight. After the reaction was completed as monitored by TLC, EA (200.0 mL) was added to the system, followed by washing with water (100.0 mL×3). The organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (100-200 mesh silica gel, petroleum ether:ethyl acetate=100:1 to 90:1) to give the product (1.8 g, yield: 56.7%).
Tert-butyl (R)-3-((4-(2-hydroxy-4-methylphenyl)phthalazin-1-yl)amino) piperidine-1-carboxylate (750.0 mg, 1.72 mmol, 1.0 eq.) was added to DCM (5.0 mL) and trifluoroacetic acid (5.0 mL), and the mixture was stirred at room temperature for 2 h. After the reaction was completed as monitored by TLC, the system was back-extracted with water (30.0 mL×4), and the aqueous phases were combined, followed by addition of a saturated aqueous sodium carbonate solution to adjust the pH to 8-9, and addition of DCM for extraction for 3 times. The organic phases were combined, dried over anhydrous sodium sulfate, and filtered, and the filtrate was concentrated under reduced pressure to give the product (550.0 mg, yield: 95.4%).
(R)-5-methyl-2-(4-(piperidin-3-ylamino)phthalazin-1-yl)phenol (550.0 mg, 1.64 mmol, 1.0 eq.) was added to methanol (10.0 mL), and an aqueous formaldehyde solution (37%) (160.1 mg, 1.97 mmol, 1.2 eq.) was added. The mixture was stirred at room temperature for 1 h, followed by addition of sodium cyanoborohydride (144.6 mg, 2.30 mmol, 1.4 eq.), and the resulting mixture was reacted at room temperature for 3 h. After the reaction was completed as monitored by TLC, EA (100.0 mL) and a saturated aqueous sodium bicarbonate solution were added to the system, followed by liquid separation. The organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and the crude product was purified by preparative thin-layer chromatography (DCM:MeOH=8:1) to give the product (300.0 mg, yield: 52.3%).
1HNMR (300 MHz, DMSO-d6) δ (ppm): 9.63 (s, 1H), 8.48 (s, 1H), 7.77-7.89 (m, 2H), 7.52-7.54 (m, 1H), 7.41 (s, 1H), 7.15-7.17 (m, 1H), 6.76-6.82 (m, 2H), 4.65 (s, 1H), 3.36-3.47 (m, 2H), 3.12-3.17 (m, 2H), 2.50-2.51 (m, 3H), 2.33 (s, 3H), 1.77-1.99 (m, 4H).
Molecular formula: C21H24N4O molecular weight: 348.45 LC-MS (Pos, m/z)=349.22 [M+H]+.
Step 1: Synthesis of 2-(2-methoxy-4-methylbenzoyl)-3-methylbenzoic acid and 2-(2-methoxy-4-methylbenzoyl)-6-methylbenzoic acid
1-Bromo-2-methoxy-4-methylbenzene (6.82 g, 33.91 mmol, 1.1 eq.) was dissolved in THE (50 mL), and the mixture was cooled to −50° C., followed by dropwise and slow addition of a solution of n-butyllithium in n-hexane (2.5 mol/L, 13.6 mL, 33.91 mmol, 1.1 eq.). The reaction solution was stirred at −50° C. for 30 min and then added dropwise and slowly to a solution of 4-methylisobenzofuran-1,3-dione (5 g, 30.83 mmol, 1.0 eq.) in THE (50 mL) at −60° C., and reacted at −60° C. in nitrogen atmosphere. After the completion of dropwise addition, the reaction solution was naturally warmed to room temperature and reacted for 1 h, and then poured into a dilute hydrochloric acid solution (100 mL), followed by extraction with EA (100 mL×2). The organic phase was washed with 0.2 mol/L NaOH solution (100 mL), hydrochloric acid was added to the aqueous phase to adjust the pH to 4, followed by extraction with EA (100 mL×2), and the organic phase was dried and concentrated to give a mixture (7.0 g, yield: 79.9%) of 2-(2-methoxy-4-methylbenzoyl)-3-methylbenzoic acid and 2-(2-methoxy-4-methylbenzoyl)-6-methylbenzoic acid.
A mixture (7.0 g, 24.62 mmol, 1.0 eq.) of 2-(2-methoxy-4-methylbenzoyl)-3-methylbenzoic acid and 2-(2-methoxy-4-methylbenzoyl)-6-methylbenzoic acid, potassium carbonate (5.1 g, 36.93 mmol, 1.5 eq.) and iodomethane (5.24 g, 36.93 mmol, 1.5 eq.) were dissolved in DMF (100 mL), and the mixture was reacted at room temperature for 1 h. After the reaction was completed as monitored by TLC, the reaction solution was poured into water (100 mL), followed by extraction with methyl tert-butyl ether (50 mL×3). The organic phase was washed with water (30 mL×2), dried, and concentrated to give a mixture (5.0 g, yield: 68.1%) of methyl 2-(2-methoxy-4-methylbenzoyl)-3-methylbenzoate and methyl 2-(2-methoxy-4-methylbenzoyl)-6-methylbenzoate.
A mixture (5.0 g, 16.76 mmol, 1.0 eq.) of methyl 2-(2-methoxy-4-methylbenzoyl)-3-methylbenzoate and methyl 2-(2-methoxy-4-methylbenzoyl)-6-methylbenzoate, and 85% hydrazine hydrate (1.48 g, 25.14 mmol, 1.5 eq.) were dissolved in ethanol (50 mL), and the mixture was reacted at 80° C. for 9 h. After the reaction was substantially completed as detected by LC-MS, the reaction solution was concentrated under reduced pressure, and the crude product was slurried with EtOH: MTBE=(3:1, 40 mL), followed by filtration under vacuum. The filter cake was dried to give a mixture (4.0 g, yield: 85.1%) of 4-(2-methoxy-4-methylphenyl)-5-methylphthalazin-1(2H)-one and 4-(2-methoxy-4-methylphenyl)-8-methylphthalazin-1(2H)-one.
4-(2-Methoxy-4-methylphenyl)-5-methylphthalazin-1(2H)-one, 4-(2-methoxy-4-methylphenyl)-8-methylphthalazin-1(2H)-one (4.0 g, 14.26 mmol, 1.0 eq.) and phosphorus oxychloride (4.37 g, 28.52 mmol, 2.0 eq.) were dissolved in acetonitrile (40 mL), and the mixture was reacted at 90° C. for 13.5 h. After the reaction was completed as monitored by TLC, the reaction solution was concentrated under reduced pressure, and the crude product was dissolved in water (50 mL), followed by addition of sodium carbonate to adjust the pH to 9, and extraction with EA (100 mL×2). The organic phase was dried and concentrated to give a mixture (3.6 g, yield: 84.5%) of 1-chloro-4-(2-methoxy-4-methylphenyl)-5-methylphthalazine and 4-chloro-1-(2-methoxy-4-methylphenyl)-5-methylphthalazine.
A mixture (3.6 g, 12.05 mmol, 1.0 eq.) of 1-chloro-4-(2-methoxy-4-methylphenyl)-5-methylphthalazine and 4-chloro-1-(2-methoxy-4-methylphenyl)-5-methylphthalazine, and tert-butyl (R)-3-aminopiperidine-1-carboxylate (4.83 g, 24.10 mmol, 2.0 eq.) were dissolved in DMAC (20 mL), and the resulting mixture was reacted at 120° C. for 11 h. After the reaction was completed as monitored by TLC, the reaction solution was poured into water (100 mL), and solids precipitated, followed by filtration under vacuum. The filter cake was dissolved in EA (100 mL), and washed with water (20 mL×3), and the organic phase was dried and concentrated to give a mixture (4.0 g, yield: 71.8%) of tert-butyl (R)-3-((4-(2-methoxy-4-methylphenyl)-5-methylphthalazin-1-yl)amino)piperidine-1-carboxylate and tert-butyl (R)-3-((4-(2-methoxy-4-methylphenyl)-8-methylphthalazin-1-yl)amino)piperidine-1-carboxylate.
A mixture (4.0 g, 8.64 mmol, 1.0 eq.) of tert-butyl (R)-3-((4-(2-methoxy-4-methylphenyl)-5-methylphthalazin-1-yl)amino)piperidine-1-carboxylate and tert-butyl (R)-3-((4-(2-methoxy-4-methylphenyl)-8-methylphthalazin-1-yl)amino)piperidine-1-carboxylate was dissolved in DCM (20 mL), and the mixture was cooled to −20° C., followed by dropwise addition of a 1 mol/L solution of boron tribromide in dichloromethane (17 mL, 17.28 mmol, 2.0 eq.). After the completion of dropwise addition, the reaction solution was naturally warmed to room temperature and reacted for 1 h, and after the reaction was completed as detected by TLC, the reaction solution was poured into water (20 mL), followed by liquid separation. NaHCO3 was added to the aqueous phase to adjust the pH to 9, and then extracted with DCM (30 mL×2), and the organic phase was dried and concentrated to give a mixture (2.5 g, yield: 83.3%) of (R)-5-methyl-2-(8-methyl-4-(piperidin-3-ylamino)phthalazin-1-yl)phenol and (R)-5-methyl-2-(5-methyl-4-(piperidin-3-ylamino)phthalazin-1-yl)phenol.
A mixture (1.0 g, 2.87 mmol, 1.0 eq.) of (R)-5-methyl-2-(8-methyl-4-(piperidin-3-ylamino)phthalazin-1-yl)phenol and (R)-5-methyl-2-(5-methyl-4-(piperidin-3-ylamino)phthalazin-1-yl)phenol was dissolved in methanol (10 mL), followed by addition of an aqueous formaldehyde solution (37%) (256 mg, 3.16 mmol, 1.1 eq.). The resulting mixture was stirred at room temperature for 20 min, followed by addition of sodium cyanoborohydride (198 mg, 3.16 mmol, 1.1 eq.). The mixture was reacted at room temperature for 20 min. After the reaction was completed as monitored by LC-MS, the reaction solution was concentrated under reduced pressure, and water (20 mL) was added to the crude product, followed by extraction with DCM (20 mL×3). The organic phase was dried and concentrated, and the crude product was purified by preparative thin-layer chromatography (DCM:MeOH=8:1) to give the product (compound 2) (R)-5-methyl-2-(8-methyl-4-((1-methylpiperidin-3-yl)amino)phthalazin-1-yl)phenol having a smaller Rf value (70 mg, yield: 6.7%).
1HNMR (400 MHz, DMSO-d6) δ (ppm): 9.31 (s, 1H), 8.31 (s, 1H), 7.74-7.72 (d, 1H), 7.60-7.58 (d, 1H), 7.31 (s, 1H), 7.06-7.03 (m, 1H), 6.73 (s, 2H), 4.68 (s, 1H), 2.57 (s, 4H), 2.50 (s, 3H), 2.31 (s, 3H), 2.06-1.99 (t, 5H), 1.83-1.81 (m, 2H).
Molecular formula: C21H26N4O molecular weight: 362.48 LC-MS (Pos, m/z)=363.26 [M+H]+.
Simultaneously, the product (compound 5) (R)-5-methyl-2-(5-methyl-4-((1-methylpiperidin-3-yl)amino)phthalazin-1-yl)phenol with a larger Rf value (40 mg, yield: 3.8%) was also given.
1HNMR (400 MHz, DMSO-d6) δ (ppm): 9.51 (s, 1H), 7.63-7.61 (d, 2H), 7.34-7.32 (t, 1H), 7.13-7.11 (d, 1H), 6.81 (s, 1H), 6.77-6.75 (d, 1H), 6.17 (s, 1H), 4.57 (s, 1H), 2.97 (s, 4H), 2.58 (s, 3H), 2.50 (s, 3H), 2.30 (s, 3H), 1.98-1.96 (d, 2H), 1.77 (s, 2H).
Molecular formula: C21H26N4O molecular weight: 362.48 LC-MS (Pos, m/z)=363.29 [M+H]+.
1-Bromo-2-methoxy-4-methylbenzene (13.64 g, 67.84 mmol, 1.1 eq.) was dissolved in THE (50 mL), the mixture was cooled to −50° C., followed by dropwise and slow addition of a solution (2.5 mol/L, 27 mL, 67.84 mmol, 1.1 eq.) of n-butyllithium in n-hexane, and the resulting mixture was stirred at −50° C. for 30 min. A solution of 5-methylisobenzofuran-1,3-dione (10 g, 61.68 mmol, 1.0 eq.) in THE (100 mL) was added dropwise and slowly to the above solution at −60° C., and the reaction solution was reacted at −60° C. for 30 min in nitrogen atmosphere. After the completion of dropwise addition, the reaction solution was naturally warmed to room temperature and reacted for 3 h, and poured into a dilute hydrochloric acid solution (200 mL), followed by addition of EA (100 mL×2) for extraction. The organic phase was washed with 0.2 mol/L NaOH solution (100 mL×2), and hydrochloric acid was added to the aqueous phase to adjust the pH to 4, followed by extraction with EA (100 mL×2). The organic phase was dried and concentrated to give a mixture (17.51 g, yield: 100%) of products.
A mixture (17.54 g, 61.68 mmol, 1.0 eq.) of 2-(2-methoxy-4-methylbenzoyl)-5-methylbenzoic acid and 2-(2-methoxy-4-methylbenzoyl)-4-methylbenzoic acid, potassium carbonate (12.83 g, 92.97 mmol, 1.5 eq.) and iodomethane (13.20 g, 92.97 mmol, 1.5 eq.) were dissolved in DMF (100 mL), and the mixture was reacted at room temperature for 30 min. After the reaction was completed as monitored by TLC, the reaction solution was poured into water (200 mL), followed by extraction with methyl tert-butyl ether (150 mL×3). The organic phase was washed with a saturated aqueous NaCl solution (100 mL×2), dried, and concentrated to give a mixture (15.0 g, yield: 82.8%) of methyl 2-(2-methoxy-4-methylbenzoyl)-5-methylbenzoate and methyl 2-(2-methoxy-4-methylbenzoyl)-4-methylbenzoate.
A mixture (15 g, 50.28 mmol, 1.0 eq.) of methyl 2-(2-methoxy-4-methylbenzoyl)-5-methylbenzoate and methyl 2-(2-methoxy-4-methylbenzoyl)-4-methylbenzoate, and 85% hydrazine hydrate (4.44 g, 75.42 mmol, 1.5 eq.) were dissolved in ethanol (150 mL), and the mixture was reacted at 80° C. for 6 h. After the reaction was completed as detected by LC-MS, the reaction solution was cooled to room temperature and filtered, and the filter cake was dried at 50° C. to give a mixture (11.0 g, yield: 78.0%) of 4-(2-methoxy-4-methylphenyl)-7-methylphthalazin-1(2H)-one and 4-(2-methoxy-4-methylphenyl)-6-methylphthalazin-1(2H)-one.
Step 4: Synthesis of 4-chloro-1-(2-methoxy-4-methylphenyl)-6-methylphthalazine and 1-chloro-4-(2-methoxy-4-methylphenyl)-6-methylphthalazine
The mixture (11.0 g, 39.24 mmol, 1.0 eq.) of 4-(2-methoxy-4-methylphenyl)-7-methylphthalazin-1(2H)-one and 4-(2-methoxy-4-methylphenyl)-6-methylphthalazin-1(2H)-one and phosphorus oxychloride (12.0 g, 78.48 mmol, 2.0 eq.) were dissolved in acetonitrile (100 mL), and the resulting mixture was reacted at 90° C. for 2 h. After the reaction was completed as monitored by TLC, the reaction solution was concentrated under reduced pressure, water (50 mL) was added, and EA (100 mL×2) was added for extraction. The organic phase was dried and concentrated to give a mixture (10.0 g, yield: 85.3%) of 4-chloro-1-(2-methoxy-4-methylphenyl)-6-methylphthalazine and 1-chloro-4-(2-methoxy-4-methylphenyl)-6-methylphthalazine.
The mixture (2.0 g, 6.69 mmol, 1.0 eq.) of 4-chloro-1-(2-methoxy-4-methylphenyl)-6-methylphthalazine and 1-chloro-4-(2-methoxy-4-methylphenyl)-6-methylphthalazine and tert-butyl (R)-3-aminopiperidine-1-carboxylate (2.68 g, 13.38 mmol, 2.0 eq.) were dissolved in DMAC (20 mL), and the resulting mixture was reacted at 120° C. for 19 h. After no materials were left as monitored by TLC, the reaction solution was poured into water (100 mL) and solids precipitated. The mixture was filtered under vacuum, and the filter cake was dried at 60° C. to give a mixture (2.0 g, yield: 64.7%) of tert-butyl (R)-3-((4-(2-methoxy-4-methylphenyl)-7-methylphthalazin-1-yl)amino)piperidine-1-carboxylate and tert-butyl (R)-3-((4-(2-methoxy-4-methylphenyl)-6-methylphthalazin-1-yl)amino)piperidine-1-carboxylate.
The mixture (2.0 g, 4.32 mmol, 1.0 eq.) of tert-butyl (R)-3-((4-(2-methoxy-4-methylphenyl)-7-methylphthalazin-1-yl)amino)piperidine-1-carboxylate and tert-butyl (R)-3-((4-(2-methoxy-4-methylphenyl)-6-methylphthalazin-1-yl)amino)piperidine-1-carboxylate was dissolved in DCM (20 mL), and the resulting mixture was cooled to −20° C., followed by dropwise addition of a 1 mol/L solution of boron tribromide in dichloromethane (21.6 mL, 21.60 mmol, 5.0 eq.). After the completion of dropwise addition, the mixture was naturally warmed to room temperature and reacted for 2 h. After the reaction was completed as detected by LC-MS, an appropriate amount of methanol was added into the flask to quench the reaction. The reaction solution was stirred for 10 min and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (DCM:MeOH=15:1) to give a mixture (1.2 g, yield: 80.0%) of (R)-5-methyl-2-(6-methyl-4-(piperidin-3-ylamino)phthalazin-1-yl)phenol and (R)-5-methyl-2-(7-methyl-4-(piperidin-3-ylamino)phthalazin-1-yl)phenol.
The mixture (1.2 g, 3.44 mmol, 1.0 eq.) of (R)-5-methyl-2-(6-methyl-4-(piperidin-3-ylamino)phthalazin-1-yl)phenol and (R)-5-methyl-2-(7-methyl-4-(piperidin-3-ylamino)phthalazin-1-yl)phenol was dissolved in methanol (15 mL), followed by addition of an aqueous formaldehyde solution (37%) (838 mg, 10.32 mmol, 3.0 eq.). The resulting mixture was stirred at room temperature for 30 min, followed by addition of sodium cyanoborohydride (648 mg, 10.32 mmol, 3.0 eq.). The mixture was reacted at room temperature for 30 min. After the reaction was completed as monitored by LC-MS, the reaction solution was concentrated under reduced pressure. Water (20 mL) was added to the crude product, and DCM (20 mL×3) was added for extraction. The organic phase was dried and concentrated to give the crude product (1.0 g). 300 mg of the crude product was purified by preparative thin-layer chromatography (dichloromethane:methanol:ammonia solution=20:1:0.5) to give the product (compound 4) (R)-5-methyl-2-(6-methyl-4-((1-methylpiperidin-3-yl)amino)phthalazin-1-yl)phenol with a larger Rf value (35 mg, yield: 9.4%).
1HNMR (400 MHz, DMSO-d6) δ (ppm): 9.68 (s, 1H), 8.18 (s, 1H), 7.61-7.58 (d, 1H), 7.46-7.43 (d, 1H), 7.17-7.15 (d, 1H), 6.91-6.89 (d, 1H), 6.79-6.75 (t, 2H), 4.40-4.38 (t, 1H), 3.08-3.06 (d, 1H), 2.72-2.69 (d, 1H), 2.52 (s, 3H), 2.32 (s, 3H), 2.21 (s, 3H), 1.99-1.92 (m, 3H), 1.75-1.74 (d, 1H), 1.65-1.56 (m, 1H), 1.49-1.43 (m, 1H).
Molecular formula: C21H26N4O Molecular weight: 362.48 LC-MS (Pos, m/z)=363.32[M+H]+.
Simultaneously, the product (compound 3) (R)-5-methyl-2-(7-methyl-4-((1-methylpiperidin-3-yl)amino)phthalazin-1-yl)phenol with a smaller Rf value (100 mg, yield: 26.8%) was also given.
1HNMR (400 MHz, DMSO-d6) δ (ppm): 9.62 (s, 1H), 8.29-8.27 (d, 1H), 7.68-7.66 (d, 1H), 7.30 (s, 1H), 7.16-7.14 (d, 1H), 7.00-6.99 (d, 1H), 6.80 (s, 1H), 6.77-6.75 (d, 1H), 4.43-4.42 (d, 1H), 3.13-3.15 (d, 1H), 2.79-2.76 (d, 1H), 2.42 (s, 3H), 2.33 (s, 3H), 2.28 (s, 3H), 2.03-1.97 (m, 3H), 1.80-1.76 (t, 1H), 1.64-1.61 (d, 1H), 1.50-1.47 (d, 1H).
Molecular formula: C21H26N4O Molecular weight: 362.48 LC-MS (Pos, m/z)=363.32[M+H]+.
1-Bromo-2-methoxy-4-methylbenzene (4.87 g, 24.22 mmol, 1.1 eq.) was dissolved in THE (50 mL), and the mixture was cooled to −60° C., followed by dropwise and slow addition of a solution of n-butyllithium in n-hexane (2.5 mol/L, 9.7 mL, 24.22 mmol, 1.1 eq.). The reaction solution was stirred at −60° C. for 30 min and then added dropwise to a solution of 4-bromoisobenzofuran-1,3-dione (5 g, 22.02 mmol, 1.0 eq.) in THE (50 mL) at −60° C. in nitrogen atmosphere. After the completion of dropwise addition, the resulting mixture was naturally warmed to room temperature and reacted for 4 h. After the reaction was completed as detected by LC-MS, the reaction solution was added into a dilute hydrochloric acid solution (100 mL), and EA (50 mL×2) was added for extraction. The organic phase was washed with a 0.2 mol/L NaOH solution (50 mL×2), and the pH of the aqueous phase was adjusted to 4 with hydrochloric acid, and EA (100 mL×2) was added for extraction. The organic phase was dried and concentrated to give the product (5.0 g, yield: 65.1%).
2-Bromo-6-(2-methoxy-4-methylbenzoyl)benzoic acid (5.0 g, 14.32 mmol, 1.0 eq.), potassium carbonate (2.96 g, 21.48 mmol, 1.5 eq.) and iodomethane (3.05 g, 21.48 mmol, 1.5 eq.) were dissolved in DMF (50 mL), and the mixture was reacted at room temperature for 0.5 h. After the reaction was completed as monitored by LC-MS, the reaction solution was poured into water (100 mL), and methyl tert-butyl ether (100 mL×2) was added for extraction. The organic phase was washed with water (50 mL×2), dried and concentrated to give the product (4.0 g, yield: 76.9%).
Methyl 2-bromo-6-(2-methoxy-4-methylbenzoyl)benzoate (4.0 g, 11.01 mmol, 1.0 eq.) and 85% hydrazine hydrate (973 mg, 16.52 mmol, 1.5 eq.) were dissolved in ethanol (40 mL), and the mixture was reacted at 80° C. for 15 h. A large number of solids precipitated, and the mixture was filtered. The filter cake was dried to give the product (1.4 g, yield: 36.8%).
Methyl 2-bromo-6-(2-methoxy-4-methylbenzoyl)benzoate (800 mg, 2.32 mmol, 1.0 eq.) and phosphorus oxychloride (711 mg, 28.52 mmol, 2.0 eq.) were dissolved in acetonitrile (10 mL), and the mixture was reacted at 90° C. for 1 h. After the reaction was completed as monitored by TLC, the reaction solution was concentrated under reduced pressure, the crude product was dissolved in EA (20 mL), and water (10 mL) was added. The mixture was stirred for 5 min, followed by liquid separation. The aqueous phase was extracted with EA (20 mL), and the organic phases were combined, dried and concentrated under reduced pressure to give the product (600 mg, yield: 71.2%).
5-bromo-4-chloro-1-(2-methoxy-4-methylphenyl)phthalazine (600 mg, 1.65 mmol, 1.0 eq.) and tert-butyl (R)-3-aminopiperidine-1-carboxylate (660 mg, 3.30 mmol, 2.0 eq.) were dissolved in DMAC (5 mL), and the mixture was reacted at 120° C. for 20 h. After the reaction was completed as monitored by TLC, the reaction solution was cooled to room temperature and poured into water (30 mL). Solids precipitated, and the mixture was filtered under vacuum. The filter cake was dried to give the product (700 mg, yield: 80.4%).
Tert-butyl (R)-3-((8-bromo-4-(2-methoxy-4-methylphenyl)phthalazin-1-yl)amino)piperidine-1-carboxylate (700 mg, 1.32 mmol, 1.0 eq.) was dissolved in DCM (10 mL), and the mixture was cooled to −20° C., followed by dropwise addition of a 1 mol/L solution of boron tribromide in dichloromethane (6.6 mL, 6.60 mmol, 5.0 eq.). After the completion of dropwise addition, the mixture was naturally warmed to room temperature and reacted for 3 h. After the reaction was completed as detected by LC-MS, an appropriate amount of methanol was added to quench the reaction. The reaction solution was concentrated under reduced pressure, and the crude product was dissolved in DCM (10 mL), followed by addition of water (10 mL) and liquid separation. The aqueous phase was extracted with DCM (10 mL×2), and the organic phases were combined, dried and concentrated to give the product (280 mg, yield: 51.4%).
(R)-2-(5-bromo-4-(piperidin-3-ylamino)phthalazin-1-yl)-5-methylphenol (250 mg, 0.60 mmol, 1.0 eq.) was dissolved in methanol (5 mL), followed by addition of an aqueous formaldehyde solution (37%) (146 mg, 1.80 mmol, 3.0 eq.). The mixture was stirred at room temperature for 30 min, followed by addition of sodium cyanoborohydride (113 mg, 1.80 mmol, 3.0 eq.). The mixture was reacted at room temperature for 30 min. After no materials were left as monitored by LC-MS, the reaction solution was concentrated. The crude product was slurried with water (10 mL) and solids precipitated. The mixture was filtered, and the filter cake was dissolved in EA (10 mL), dried and filtered. The filtrate was concentrated under reduced pressure, and the crude product was purified by preparative thin-layer chromatography (DCM:MeOH=10:1) to give the product (120 mg, yield: 46.8%).
1HNMR (300 MHz, DMSO-d6) δ (ppm): 9.50 (s, 1H), 8.15-8.13 (m, 1H), 7.66-7.61 (t, 1H), 7.50-7.47 (m, 2H), 7.16-7.13 (d, 1H), 6.80-6.76 (t, 2H), 4.53 (s, 1H), 2.80-2.75 (t, 1H), 2.58 (d, 2H), 2.33 (s, 7H), 1.79 (s, 3H), 1.65 (s, 1H).
Molecular formula: C21H23Br N4O Molecular weight: 427.35 LC-MS (Pos, m/z)=427.25/429.25[M+H]+.
3,4-Dimethylphenol (5.0 g, 40.9 mmol, 1.0 eq.) was dissolved in DCM (50 mL), and NBS (7.28 g, 40.9 mmol, 1.0 eq.) was added portionwise under an ice-water bath. The mixture was reacted for 2 h. The reaction solution was washed with water, and the organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (PE:EA=50:1 to 20:1) to give the product (7.20 g, yield: 87.5%).
2-Bromo-4,5-dimethylphenol (7.20 g, 35.8 mmol, 1.0 eq.) was dissolved in DMF (80 mL), and then K2CO3 (9.90 g, 71.6 mmol, 2.0 eq.) and CH3I (7.62 g, 53.7 mmol, 1.5 eq.) were added. The mixture was stirred at room temperature for 3 h. Water was added to quench the reaction, and EA (50 mL) was added for extraction. The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (PE:EA=50:1 to 20:1) to give the product (7.10 g, yield: 92.2%).
1-Bromo-2-methoxy-4,5-dimethylbenzene (5.00 g, 23.2 mmol, 1.0 eq.) and triisopropyl borate (8.73 g, 46.4 mmol, 2.0 eq.) were dissolved in THF (50 mL), followed by purge with nitrogen for protection. The mixture was cooled to −70° C., followed by dropwise addition of n-butyllithium (18.6 mL, 46.4 mmol, 2.0 eq.). The mixture was reacted for 3 h. Dilute hydrochloric acid was added to quench the reaction, the pH was adjusted to 2, and EA (50 mL) was added for extraction. The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure and slurried with petroleum ether to give the product (2.60 g, yield: 62.1%).
(2-Methoxy-4,5-dimethylphenyl)boronic acid (194 mg, 1.08 mmol, 1.5 eq.), (R)-4-chloro-N-(1-methylpiperidin-3-yl)phthalazin-1-amine (200 mg, 0.723 mmol, 1.0 eq.), Pd(dppf)Cl2 (26.5 mg, 0.0362 mmol, 0.05 eq.) and NaHCO3 (122 mg, 1.45 mmol, 2.0 eq.) were added successively to 1,4-dioxane (4 mL), and H2O (2 mL) was added, followed by purge with nitrogen for protection. The oil bath was heated to 110° C. and the mixture was reacted for 1 h. The reaction solution was cooled down, water was added to quench the reaction, and EA (20 mL) was added for extraction. The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (DCM:MeOH=50:1 to 20:1) to give the product (200 mg, yield: 73.5%).
(R)-4-(2-methoxy-4,5-dimethylphenyl)-N-(1-methylpiperidin-3-yl)phthalazin-1-amine (200 mg, 0.531 mmol, 1.0 eq.) was dissolved in DCM (4 mL), and the mixture was cooled to −10° C., followed by dropwise addition of BBr3 (266 mg, 1.06 mmol, 2.0 eq.). The mixture was reacted for 2 h. MeOH was added to quench the reaction, a saturated aqueous NaHCO3 solution was added to adjust the pH to 8, and DCM (20 mL) was added for extraction. The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by preparative thin-layer chromatography (DCM:MeOH=10:1) to give the product (120 mg, yield: 62.3%).
1HNMR (300 MHz, DMSO-d6) δ (ppm): 9.37 (s, 1H), 8.41 (d, J=8.1 Hz, 1H), 7.86-7.74 (m, 2H), 7.54 (d, J=7.8 Hz, 1H), 7.14 (s, 1H), 7.04 (s, 1H), 6.80 (s, 1H), 4.49 (s, 1H), 3.21 (d, J=8.4 Hz, 1H), 2.84 (d, J=11.1 Hz, 1H), 2.35 (s, 3H), 2.23-2.18 (m, 8H), 2.02-1.97 (m, 1H), 7.84-1.80 (m, 1H), 1.72-1.49 (m, 2H).
Molecular formula: C22H26N4O Molecular weight: 362.48 LC-MS (Pos, m/z)=363.34[M+H]+.
Thiophene-3,4-dicarboxylic acid (5.0 g, 29.0 mmol, 1.0 eq.) was dissolved in EtOH (50 mL), followed by addition of H2SO4 (5.69 g, 58.0 mmol, 2.0 eq.). The oil bath was heated to 80° C. and the mixture was reacted for 20 h. The reaction solution was concentrated to remove EtOH, water was added for dissolution, and EA (50 mL) was added for extraction. The organic phase was washed with a saturated aqueous NaHCO3 solution (40 mL), dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to give the product (6.20 g, yield: 93.5%).
Diethyl thiophene-3,4-dicarboxylate (6.20 g, 27.2 mmol, 1.0 eq.) was dissolved in MeOH (30 mL), followed by addition of hydrazine hydrate (85%, 8.01 g, 136 mmol, 5.0 eq.). The oil bath was heated to 65° C. and the mixture was reacted for 4 h. The reaction solution was cooled to room temperature, filtered under vacuum and dried to give the product (3.48 g, yield: 76.2%).
2,3-Dihydrothieno[3,4-d]pyridazine-1,4-dione (1.00 g, 5.95 mmol, 1.0 eq.) was added to ACN (10 mL), followed by addition of POCl3 (2.00 g, 13.1 mmol, 2.2 eq.). The oil bath was heated to 90° C. and the mixture was reacted for 1 h. The reaction solution was cooled to room temperature, water (20 mL) was added to quench the reaction, and EA (20 mL) was added for extraction. The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to give 1,4-dichlorothieno[3,4-d]pyridazine (780 mg, yield: 64.0%).
(2-Methoxy-4-methylphenyl)boronic acid (631 mg, 3.8 mmol, 1.5 eq.), 1,4-dichlorothieno[3,4-d]pyridazine (780 mg, 3.80 mmol, 1.0 eq.), Pd(dppf)Cl2 (139 mg, 0.190 mmol, 0.05 eq.) and NaHCO3 (1.28 g, 7.60 mmol, 2.0 eq.) were added successively to 1,4-dioxane (10 mL), and H2O (5 mL) was added, followed by purge with nitrogen for protection. The oil bath was heated to 110° C. and the mixture was reacted for 1 h. Water was added to quench the reaction, and EA (30 mL) was added for extraction. The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (DCM:MeOH=100:1 to 20:1) to give the product (820 mg, yield: 79.2%).
4-(2-Methoxy-4-methylphenyl)thieno[3,4-d]pyridazin-1-ol (820 mg, 3.01 mmol, 1.0 eq.) was added to ACN (10 mL), followed by addition of POCl3 (693 mg, 4.52 mmol, 1.5 eq.). The oil bath was heated to 90° C. and the mixture was reacted for 1 h. The reaction solution was cooled to room temperature, water was added to quench the reaction, and EA (20 mL) was added for extraction. The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to give the product (680 mg, yield: 77.7%).
1-Chloro-4-(2-methoxy-4-methylphenyl)thieno[3,4-d]pyridazine (600 mg, 2.06 mmol, 1.0 eq.) and tert-butyl (R)-3-aminopiperidine-1-carboxylate (619 mg, 3.09 mmol, 1.5 eq.) were dissolved in DMA (10 mL), followed by purge with nitrogen for protection. The oil bath was heated to 120° C. and the mixture was reacted for 20 h. Water was added to quench the reaction, and EA (30 mL) was added for extraction. The organic phase was washed three times with water (30 mL×3), dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to give tert-butyl (R)-3-((4-(2-methoxy-4-methylphenyl)thieno[3,4-d]pyridazin-1-yl)amino)piperidine-1-carboxylate (420 mg, yield: 44.8%).
Tert-butyl (R)-3-((4-(2-methoxy-4-methylphenyl)thieno[3,4-d]pyridazin-1-yl)amino)piperidine-1-carboxylate (420 mg, 0.924 mmol, 1.0 eq.) was dissolved in DCM (8 mL), and the mixture was cooled to −10° C., followed by dropwise addition of BBr3 (1.16 mg, 4.62 mmol, 5.0 eq.). The mixture was reacted for 2 h. MeOH was added to quench the reaction, and the reaction solution was concentrated. The crude product was dissolved in H2O and extracted with EA. A saturated aqueous NaHCO3 solution was added to the aqueous phase to adjust the pH to 8, and DCM (20 mL×3) was added for extraction. The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to give the product (182 mg, yield: 57.9%).
(R)-5-methyl-2-(4-(piperidin-3-ylamino)thieno[3,4-d]pyridazin-1-yl)phenol (180 mg, 0.529 mmol, 1.0 eq.) was dissolved in MeOH (4 mL), followed by addition of an aqueous formaldehyde solution (129 mg, 1.59 mmol, 3.0 eq.). The mixture was stirred at room temperature for 30 min. Sodium cyanoborohydride (100 mg, 1.59 mmol, 3.0 eq.) was then added and the mixture was reacted for 1 h. A saturated aqueous NaHCO3 solution was added to quench the reaction, and EA (20 mL) was added for extraction. The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was separated by preparative thin-layer chromatography to give the product (52.0 mg, yield: 27.7%).
1HNMR (300 MHz, DMSO-d6) δ (ppm): 13.28 (s, 1H), 9.01 (s, 1H), 8.66 (d, J=2.7 Hz, 1H), 7.87 (d, J=7.8 Hz, 2H), 8.49 (d, J=8.4 Hz, 2H), 4.59 (s, 1H), 3.11 (s, 2H), 2.74 (s, 1H), 2.64 (s, 3H), 2.32 (s, 3H), 1.99-1.74 (m, 5H).
Molecular formula: C19H22N4OS Molecular weight: 354.47 LC-MS (Pos, m/z)=355.19[M+H]+.
1-Bromo-2-methoxy-4-methylbenzene (14.15 g, 70.38 mmol, 1.05 eq.) was dissolved in anhydrous THE (100 mL), followed by dropwise addition of n-BuLi (33.8 mL, 80.48 mmol, 1.2 eq.) at −78° C. After the completion of dropwise addition, the mixture was reacted for 30 min. At −78° C., the reaction solution was added dropwise to a solution of 2,3-pyridine dicarboxylic anhydride (10 g, 67.07 mmol, 1.0 eq.) in THE (100 mL). After the completion of dropwise addition, the mixture was reacted at room temperature for 16 h. After the reaction was completed as detected by LC-MS, the reaction solution was poured into a mixed solution of ice water (50 mL) and 2 N HCl (50 mL), and ethyl acetate (200 mL) was added for extraction. The aqueous phase was extracted with dichloromethane (200 mL), and the organic phases were combined, dried and concentrated to give the crude product (26 g). The crude product was subjected to silica gel column chromatography (petroleum ether:ethyl acetate=2:1) to give the product (1.9 g, yield: 10.4%).
The 2-(2-methoxy-4-methylbenzoyl)nicotinic acid (1.9 g, 7 mmol, 1.0 eq.) obtained in the previous step was dissolved in DMF (15 mL), followed by addition of potassium carbonate (1.45 g, 10.5 mmol, 1.5 eq.) and iodomethane (1.19 g, 8.4 mmol, 1.2 eq.). The mixture was reacted at room temperature for 16 h. After the reaction was completed as detected by TLC, the reaction solution was poured into water (20 mL), and EA (20 mL) was added for extraction. The organic phase was washed with water (20 mL) and saturated brine (20 mL), dried over anhydrous sodium sulfate and concentrated to give the crude product (1.4 g). The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=5:1 to 3:1) to give the product (1.23 g, yield: 61.5%).
Methyl 2-(2-methoxy-4-methylbenzoyl)nicotinate (1.23 g, 4.31 mmol, 1.0 eq.) obtained in the previous step was dissolved in ethanol (10 mL), followed by addition of 85% hydrazine hydrate (324 mg, 4.47 mmol, 1.5 eq.). The mixture was heated to 85° C. and reacted for 2 h. After the reaction was completed as detected by TLC, the reaction solution was cooled to room temperature, and the product precipitated. The mixture was filtered to give the product (1.02 g, yield: 88.6%).
Acetonitrile (10 mL) was added to 8-(2-methoxy-4-methylphenyl)pyrido[2,3-d]pyridazin-5-ol (1.02 g, 3.82 mmol, 1.0 eq.), followed by addition of phosphorus oxychloride (1.17 g, 7.64 mmol, 2.0 eq.). The mixture was stirred at 90° C. for 6 h. After the reaction was completed as detected by TLC, the reaction solution was concentrated. The concentrated solution was poured into ice water (10 mL), and ethyl acetate (10 mL) was added for extraction. The organic phase was washed with water (10 mL) and saturated brine (10 mL), dried and concentrated to give the product (950 mg, yield: 87%).
5-Chloro-8-(2-methoxy-4-methylphenyl)pyrido[2,3-d]pyridazine (950 mg, 3.325 mmol, 1.0 eq.) was dissolved in DMA (10 mL), followed by addition of (R)-1-tert-butoxycarbonyl-3-aminopiperidine (1.33 g, 6.65 mmol, 2.0 eq.). The mixture was refluxed at 120° C. for 64 h. After the reaction was completed as detected by TLC, the reaction solution was added dropwise into water (10 mL), and ethyl acetate (10 mL) was added for extraction. The organic phase was washed with water (10 mL) and saturated brine (10 mL), dried and concentrated, and the crude product was subjected to silica gel column chromatography (petroleum ether:ethyl acetate=1.5:1 to only ethyl acetate) to give the product (805 mg, yield: 54%).
Tert-butyl (R)-3-((8-(2-methoxy-4-methylphenyl)pyrido[2,3-d]pyridazin-5-yl)amino)piperidine-1-carboxylate (805 mg, 1.79 mmol, 1.0 eq.) was dissolved in dichloromethane (5 mL), followed by dropwise addition of boron tribromide (1.35 g, 5.37 mmol, 3.0 eq.) at −20° C. The mixture was reacted at room temperature for 1 h. After the reaction was not completed as detected by LC-MS, the reaction solution was supplemented with boron tribromide (1.35 g, 5.37 mmol, 3.0 eq.). The resulting mixture was reacted at room temperature for 1 h. After the reaction was completed as detected by LC-MS, methanol (1 mL) and water (10 mL) were added dropwise and slowly to the reaction solution under an ice bath, followed by extraction with dichloromethane (10 mL). The organic phase was dried and concentrated to give the product (146 mg, yield: 24.3%).
(R)-5-Methyl-2-(5-((piperidin-3-yl)amino)pyrido[2,3-d]pyridazin-8-yl)phenol (146 mg, 0.435 mmol, 1.0 eq.) was dissolved in methanol (2 mL), followed by addition of an aqueous formaldehyde solution (38.5%) (34 mg, 0.435 mmol, 1.0 eq.). The mixture was stirred for 5 min, followed by addition of sodium cyanoborohydride (30 mg, 0.479 mmol, 1.1 eq.). The resulting mixture was reacted at room temperature for 10 min. After the reaction was completed as detected by TLC, the reaction solution was concentrated, and the crude product was purified by preparative thin-layer chromatography (dichloromethane:methanol=7:1) to give the product (34 mg, yield: 22.3%).
1H-NMR (300 MHz, DMSO-d6) δ (ppm): 11.92 (brs, 1H), 9.20-9.22 (m, 1H), 9.01 (s, 1H), 8.18-8.21 (d, 1H), 7.95-7.99 (m, 1H), 7.84 (s, 1H), 6.77 (s, 1H), 6.73 (s, 1H), 4.66 (s, 1H), 3.55 (s, 3H), 2.72 (s, 4H), 2.31 (s, 3H), 1.99-2.02 (d, 2H), 1.79-1.83 (d, 2H)
Molecular formula: C20H23N5O Molecular weight: 349.44 LC-MS: 350.2 [M+H]+.
1-Bromo-2-methoxy-4-methylbenzene (14.15 g, 70.38 mmol, 1.05 eq.) was dissolved in anhydrous THE (100 mL), followed by dropwise addition of n-BuLi (33.8 mL, 80.48 mmol, 1.2 eq.) at −78° C. After the completion of dropwise addition, the mixture was reacted for 30 min. At −78° C., the reaction solution was added dropwise to a solution of 2,3-pyridine dicarboxylic anhydride (10 g, 67.07 mmol, 1.0 eq.) in THE (100 mL). After the completion of dropwise addition, the mixture was reacted at room temperature for 16 h. After the reaction was completed as detected by LC-MS, the reaction solution was poured into a mixed solution of ice water (50 mL) and 2 N HCl (50 mL), and ethyl acetate (200 mL) was added for extraction. The aqueous phase was extracted with dichloromethane (200 mL), and the organic phases were combined, dried and concentrated to give the crude product (26 g). The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate=2:1, dichloromethane:methanol=10:1) to give the product (1 g, yield: 5.5%).
The 3-(2-methoxy-4-methylbenzoyl)nicotinic acid (1 g, 3.69 mmol, 1.0 eq.) obtained in the previous step was dissolved in DMF (15 mL), followed by addition of potassium carbonate (766 mg, 5.54 mmol, 1.5 eq.) and iodomethane (629 mg, 4.43 mmol, 1.2 eq.). The mixture was reacted at room temperature for 16 h. After the reaction was completed as detected by TLC, the reaction solution was poured into water (20 mL), and EA (20 mL) was added for extraction. The organic phase was washed with water (20 mL) and saturated brine (20 mL), dried over anhydrous sodium sulfate and concentrated to give the product (456 mg, yield: 43.4%).
Methyl 3-(2-methoxy-4-methylbenzoyl)nicotinate (456 mg, 1.6 mmol, 1.0 eq.) obtained in the previous step was dissolved in ethanol (10 mL), followed by addition of 85% hydrazine hydrate (142 mg, 2.4 mmol, 1.5 eq.). The mixture was heated to 85° C. and reacted for 2 h. After the reaction was completed as detected by TLC, the reaction solution was concentrated. Water (10 mL) was added and dichloromethane (10 mL) was added for extraction. The organic phase was dried and concentrated to give the product (230 mg, yield: 53.8%).
Acetonitrile (3 mL) was added to 5-(2-methoxy-4-methylphenyl)pyrido[2,3-d]pyridazin-8-ol (230 mg, 0.86 mmol, 1.0 eq.), followed by addition of phosphorus oxychloride (264 mg, 1.72 mmol, 2.0 eq.). The mixture was reacted at 85° C. for 6 h. After the reaction was completed as detected by TLC, the reaction solution was concentrated. The concentrated solution was poured into ice water (10 mL), and dichloromethane (10 mL) was added for extraction. The organic phase was washed with water (10 mL) and saturated brine (10 mL), dried and concentrated to give the product (112 mg, yield: 45.7%).
8-Chloro-5-(2-methoxy-4-methylphenyl)pyrido[2,3-d]pyridazine (112 mg, 0.392 mmol, 1.0 eq.) was dissolved in DMA (2 mL), followed by addition of (R)-1-tert-butoxycarbonyl-3-aminopiperidine (157 mg, 0.784 mmol, 2.0 eq.). The mixture was refluxed at 120° C. for 64 h. After the reaction was completed as detected by TLC, the reaction solution was added dropwise into water (5 mL), and ethyl acetate (5 mL) was added for extraction. The organic phase was washed with water (5 mL) and saturated brine (5 mL), dried and concentrated to give the crude product (280 mg).
Tert-butyl (R)-3-((5-(2-methoxy-4-methylphenyl)pyrido[2,3-d]pyridazin-8-yl)amino)piperidine-1-carboxylate (280 mg, crude product, 0.392 mmol, 1.0 eq.) was dissolved in dichloromethane (2 mL), followed by dropwise addition of boron tribromide (295 mg, 1.176 mmol, 3.0 eq.) at −20° C. The mixture was reacted at room temperature for 1 h. After the reaction was not completed as detected by LC-MS, the reaction solution was supplemented with boron tribromide (295 mg, 1.176 mmol, 3.0 eq.). The resulting mixture was reacted at room temperature for 1 h. After the reaction was completed as detected by LC-MS, methanol (5 mL) and water (10 mL) were added dropwise and slowly to the reaction solution under an ice bath. Dichloromethane (10 mL) was added for extraction, and the product was in the aqueous phase. The pH of the aqueous phase was adjusted to be neutral with a saturated aqueous sodium bicarbonate solution, and dichloromethane (10 mL) was added for extraction. The organic phase was dried and concentrated to give the product (70 mg, two-step yield: 53%).
(R)-8-Methyl-2-(5-((piperidin-3-yl)amino)pyrido[2,3-d]pyridazin-5-yl)phenol (70 mg, 0.209 mmol, 1.0 eq.) was dissolved in methanol (1 mL), followed by addition of an aqueous formaldehyde solution (38.5%) (16.3 mg, 0.209 mmol, 1.0 eq.). The mixture was stirred for 5 min, followed by addition of sodium cyanoborohydride (14.5 mg, 0.230 mmol, 1.1 eq.). The resulting mixture was reacted at room temperature for 10 min. After the reaction was completed as detected by TLC, the reaction solution was concentrated, and the crude product was purified by preparative thin-layer chromatography (dichloromethane:methanol=7:1) to give the product (17 mg, yield: 23.2%).
1H-NMR (300 MHz, DMSO-d6) δ (ppm): 9.71 (s, 1H), 9.11-9.13 (m, 1H), 7.85-7.98 (m, 2H), 7.54 (s, 1H), 7.22-7.24 (d, 1H), 6.79-6.83 (m, 2H), 4.59 (s, 1H), 2.92-3.04 (m, 2H), 2.74 (m, 2H), 2.59 (s, 3H), 2.34 (s, 3H), 1.78-1.92 (m, 4H).
Molecular formula: C20H23N5O Molecular weight: 349.44 LC-MS: 350.2 [M+H]+.
1-Bromo-4-methyl-2-nitrobenzene (20.0 g, 92.57 mmol, 1.0 eq.) was added into ethanol (200.0 mL), followed by addition of a saturated aqueous ammonium chloride solution (30.0 mL) and reduced iron powder (51.8 g, 925.7 mmol, 10.0 eq.). The mixture was stirred at 80° C. for 4 h. After the reaction was completed as monitored by TLC, the system was cooled to room temperature and filtered under vacuum. The filtrate was concentrated under reduced pressure, and dichloromethane (200.0 mL) was added, followed by washing with water (100.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (specification of silica gel: 100-200 mesh, petroleum ether:ethyl acetate=20:1) to give the product (16.0 g, yield: 92.9%).
2-Bromo-5-methylaniline (16.0 g, 85.99 mmol, 1.0 eq.), bis(pinacolato)diboron (32.7 g, 128.99 mmol, 1.5 eq.), potassium acetate (16.8 g, 171.98 mmol, 2.0 eq.) and Pd(dppf)Cl2 (3.1 g, 4.29 mmol, 0.05 eq.) were added to 1,4-dioxane (200.0 mL), and the mixture was reacted at 100° C. for 3 h in nitrogen atmosphere. After the reaction was completed as monitored by TLC, the system was concentrated, and the crude product was purified by silica gel column chromatography (specification of silica gel: 100-200 mesh, petroleum ether:ethyl acetate=50:1 to 30:1) to give the product (15.0 g, yield: 75.0%).
5-Methyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (10.0 g, 42.89 mmol, 1.0 eq.), di-tert-butyl dicarbonate (18.7 g, 85.79 mmol, 2.0 eq.) and 4-dimethylaminopyridine (7.85 g, 64.33 mmol, 1.5 eq.) were added to dichloromethane (150.0 mL), and the mixture was reacted at room temperature for 3 h. After the reaction was completed as monitored by TLC, dichloromethane (300.0 mL) was added to the system, followed by washing with a saturated aqueous ammonium chloride solution (200.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (specification of silica gel: 100-200 mesh, petroleum ether:ethyl acetate=100:1 to 80:1 to 60:1) to give the product (8.0 g, yield: 57.1%).
1,4-Dichlorophthalazine (3.67 g, 18.47 mmol, 1.0 eq.) and tert-butyl (R)-3-aminopiperidine-1-carboxylate (7.4 g, 36.94 mmol, 2.0 eq.) were added to N,N-dimethylacetamide (15.0 mL), and the mixture was stirred at 120° C. for 12 h. After the reaction was completed as monitored by TLC, ethyl acetate (200.0 mL) was added to the system, followed by washing with water (100.0 mL×3). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (specification of silica gel: 100-200 mesh, petroleum ether:ethyl acetate=10:1 to 2:1) to give the product (1.5 g, yield: 22.4%).
Tert-butyl (R)-3-((4-chlorophthalazin-1-yl)amino)piperidine-1-carboxylate (1.5 g, 4.13 mmol, 1.0 eq.) and trifluoroacetic acid (6.0 mL) were added to dichloromethane (6.0 mL), and the mixture was stirred at room temperature for 2 h. After the reaction was completed as monitored by TLC, the system was back-extracted with water (100.0 mL), and the pH of the aqueous phase was adjusted to 8-9, followed by addition of dichloromethane (100.0 mL×3) for extraction. The organic phases were combined, dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to give the product (1.0 g, yield: 92.1%).
(R)-4-chloro-N-(piperidin-3-yl)phthalazin-1-amine (1.0 g, 3.80 mmol, 1.0 eq.), dichloromethane (3.0 mL) and an aqueous formaldehyde solution (37%) (308.9 mg, 3.80 mmol, 1.0 eq.) were added to methanol (10.0 mL), and the mixture was stirred at room temperature for 1 h, followed by addition of sodium cyanoborohydride (286.5 mg, 4.56 mmol, 1.2 eq.). The resulting mixture was reacted at room temperature for 1 h. After the reaction was completed as monitored by TLC, the system was concentrated under reduced pressure, followed by addition of a saturated aqueous sodium bicarbonate solution (100.0 mL). The system was stirred for 30 min and extracted with dichloromethane (100.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (specification of silica gel: 100-200 mesh, dichloromethane:methanol=50:1 to 30:1 to 10:1) to give the product (820.0 mg, yield: 78.0%).
(R)-4-chloro-N-(1-methylpiperidin-3-yl)phthalazin-1-amine (274.0 mg, 0.98 mmol, 1.0 eq.), tert-butyl (5-methyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)carbamate (489.8 mg, 1.47 mmol, 1.5 eq.), sodium bicarbonate (164.6 mg, 1.96 mmol, 2.0 eq.), water (3.0 mL) and Pd(PPh3)4 (113.2 mg, 0.098 mmol, 0.1 eq.) were added to 1,4-dioxane (8.0 mL), and the mixture was reacted at 110° C. for 4 h in nitrogen atmosphere. After the reaction was completed as monitored by TLC, the system was cooled to room temperature and filtered, and ethyl acetate (100.0 mL) was added to the filtrate, followed by washing with water (100.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by preparative thin-layer chromatography (dichloromethane:methanol=10:1) to give the product (201.0 mg, yield: 45.8%).
Tert-butyl (R)-(5-methyl-2-(4-((1-methylpiperidin-3-yl)amino)phthalazin-1-yl)phenyl)carbamate (201.0 mg, 0.44 mmol, 1.0 eq.) and trifluoroacetic acid (3.0 mL) were added to dichloromethane (3.0 mL), and the mixture was reacted at room temperature for 1 h. After the reaction was completed as monitored by TLC, the system was back-extracted with water (50.0 mL), the pH of the aqueous phase was adjusted to 8-9, and dichloromethane (50.0 mL×2) was added for extraction. The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to give the product (120.0 mg, yield: 76.9%).
1HNMR (300 MHz, DMSO-d6) δ (ppm): 8.37-8.40 (d, J=9 Hz, 1H), 7.82-7.87 (m, 2H), 7.55-7.58 (d, J=9 Hz, 1H), 7.02-7.05 (d, J=9 Hz, 1H), 6.93-6.96 (d, J=9 Hz, 1H), 6.65 (s, 1H), 6.50-6.53 (d, J=9 Hz, 1H), 4.89 (s, 2H), 4.41-4.43 (d, J=6 Hz, 1H), 3.08-3.10 (d, J=6 Hz, 1H), 2.70-2.73 (d, J=9 Hz, 1H), 2.27 (s, 3H), 2.22 (s, 3H), 1.40-1.99 (m, 6H).
Molecular formula: C21H25N5 Molecular weight: 347.47 LC-MS (Pos, m/z)=348.25[M+H]+.
2-Bromo-5-methylaniline (5.0 g, 26.87 mmol, 1.0 eq.) was added to tetrahydrofuran (20.0 mL), and the mixture was cooled to −70° C. in nitrogen atmosphere, followed by dropwise addition of a solution of n-butyllithium in n-hexane (2.5 M) (10.7 mL, 26.87 mmol, 1.0 eq.). The resulting mixture was reacted for 1 h and then warmed to room temperature, followed by dropwise addition of iodomethane (3.8 g, 26.87 mmol, 1.0 eq.). The resulting mixture was reacted at room temperature for 3 h. After the reaction was completed as monitored by TLC, ethyl acetate (200.0 mL) was added to the system, followed by washing with a saturated aqueous ammonium chloride solution (100.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (specification of silica gel: 100-200 mesh, petroleum ether:ethyl acetate=100:1) to give the product (4.7 g, yield: 87.4%).
2-Bromo-N,5-dimethylaniline (1.0 g, 4.99 mmol, 1.0 eq.), bis(pinacolato)diboron (1.9 g, 7.49 mmol, 1.5 eq.), potassium acetate (979.4 mg, 9.98 mmol, 2.0 eq.) and Pd(PPh3)4 (288.3 mg, 0.24 mmol, 0.05 eq.) were added to 1,4-dioxane (10.0 mL), followed by purge with nitrogen for 5 min. The mixture was reacted at 100° C. for 12 h. After the reaction was completed as monitored by TLC, the reaction solution was concentrated and purified by silica gel column chromatography (specification of silica gel: 100-200 mesh, petroleum ether:ethyl acetate=1:1) to give the product (800.0 mg, yield: 64.3%).
(R)-4-chloro-N-(1-methylpiperidin-3-yl)phthalazin-1-amine (160.0 mg, 0.57 mmol, 1.0 eq.), N,5-dimethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (211.3 mg, 0.85 mmol, 1.5 eq.), sodium bicarbonate (95.7 mg, 1.14 mmol, 2.0 eq.), water (2.0 mL) and Pd(PPh3)4 (32.9 mg, 0.02 mmol, 0.05 eq.) were added to 1,4-dioxane (6.0 mL), and the mixture was reacted at 110° C. for 12 h in nitrogen atmosphere. After the reaction was completed as monitored by TLC, the system was cooled to room temperature and filtered, and ethyl acetate (100.0 mL) was added to the filtrate, followed by washing with water (100.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by preparative thin-layer chromatography (dichloromethane:methanol=8:1) to give the product (70.0 mg, yield: 33.9%).
1HNMR (300 MHz, DMSO-d6) δ (ppm): 8.42-8.45 (d, J=9 Hz, 1H), 7.75-7.88 (m, 2H), 7.49-7.51 (d, J=6 Hz, 1H), 7.24 (s, 1H), 6.93-6.96 (d, J=9 Hz, 1H), 6.53-6.56 (m, 2H), 5.02-5.03 (d, J=3 Hz, 1H), 4.56 (s, 1H), 2.95-2.98 (d, J=9 Hz, 1H), 2.62-2.63 (d, J=3 Hz, 3H), 2.46 (s, 3H), 2.34 (s, 3H), 1.60-2.28 (m, 6H).
Molecular formula: C22H27N5 Molecular weight: 361.49 LC-MS (Pos, m/z)=362.26[M+H]+.
2-(4-Bromophthalazin-1-yl)-5-methylphenol (300.0 mg, 0.95 mmol, 1.0 eq.) and tert-butyl (R)-3-(methylamino)piperidine-1-carboxylate (407.9 mg, 1.90 mmol, 2.0 eq.) were added to N,N-dimethylacetamide (10.0 mL), and the mixture was stirred at 120° C. for 12 h. After the reaction was completed as monitored by TLC, the system was cooled to room temperature, and ethyl acetate (100.0 mL) was added, followed by washing with water (100.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (specification of silica gel: 100-200 mesh, petroleum ether:ethyl acetate=20:1 to 6:1) to give the product (160.0 mg, yield: 37.5%).
Tert-butyl (R)-3-((4-(2-hydroxy-4-methylphenyl)phthalazin-1-yl)(methyl) amino)piperidine-1-carboxylate (160.0 mg, 0.35 mmol, 1.0 eq.) and trifluoroacetic acid (5.0 mL) were added to dichloromethane (5.0 mL), and the mixture was stirred at room temperature for 2 h. After the reaction was completed as monitored by TLC, the system was back-extracted with water (80.0 mL), the pH of the aqueous phase was adjusted to 8-9, and dichloromethane (100.0 mL×2) was added for extraction. The organic phases were combined, dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to give the product (98.0 mg, yield: 80.3%).
(R)-5-methyl-2-(4-(methyl(piperidin-3-yl)amino)phthalazin-1-yl)phenol (98.0 mg, 0.28 mmol, 1.0 eq.) and an aqueous formaldehyde solution (37%) (22.8 mg, 0.28 mmol, 1.0 eq.) were added to methanol (5.0 mL), and the mixture was stirred at room temperature for 1 h, followed by addition of sodium cyanoborohydride (19.3 mg, 0.30 mmol, 1.1 eq.). The resulting mixture was reacted at room temperature for 2 h. After the reaction was completed as monitored by TLC, a saturated aqueous sodium bicarbonate solution (50.0 mL) was added to the system, and the mixture was stirred for 30 min, followed by extraction with dichloromethane (100.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by preparative thin-layer chromatography (dichloromethane:methanol=10:1) to give the product (30.0 mg, yield: 29.5%).
1HNMR (300 MHz, DMSO-d6) δ (ppm): 9.63 (s, 1H), 8.36-8.48 (m, 1H), 8.11-8.19 (m, 1H), 7.89-7.94 (m, 1H), 7.79-7.85 (m, 1H), 7.54-7.57 (d, J=9 Hz, 1H), 7.03 (s, 1H), 6.85-6.88 (d, J=9 Hz, 1H), 4.26 (s, 1H), 3.46-3.49 (m, 1H), 3.19-3.28 (m, 4H), 2.66 (s, 3H), 2.08 (m, 1H), 2.45 (s, 3H), 1.96-2.06 (m, 5H).
Molecular formula: C22H26N4O Molecular weight: 362.48 LC-MS (Pos, m/z)=363.33[M+H]+.
4-Bromo-3-methoxybenzonitrile (5.0 g, 23.50 mmol, 1.0 eq.), bis(pinacolato)diboron (11.97 g, 47.15 mmol, 2.0 eq.), potassium acetate (6.94 g, 70.73 mmol, 3.0 eq.) and Pd(PPh3)4 (1.36 g, 1.17 mmol, 0.05 eq.) were added to 1,4-dioxane (100.0 mL), and the mixture was reacted at 100° C. for 12 h in nitrogen atmosphere. After the reaction was completed as monitored by TLC, the system was directly concentrated, and the crude product was purified by silica gel column chromatography (specification of silica gel: 100-200 mesh, petroleum ether:ethyl acetate=20:1) to give the product (5.1 g, yield: 85.0%).
(R)-4-chloro-N-(1-methylpiperidin-3-yl)phthalazin-1-amine (154.0 mg, 0.55 mmol, 1.0 eq.), 3-methoxy-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzonitrile (288.3 mg, 1.11 mmol, 2.0 eq.), sodium bicarbonate (140.1 mg, 1.66 mmol, 3.0 eq.), water (1.0 mL) and Pd(PPh3)4 (32.1 mg, 0.027 mmol, 0.05 eq.) were added to 1,4-dioxane (3.0 mL), and the mixture was reacted at 110° C. for 3 h in nitrogen atmosphere. After the reaction was completed as monitored by TLC, the system was cooled to room temperature and filtered, and ethyl acetate (100.0 mL) was added to the filtrate, followed by washing with water (100.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (specification of silica gel: 100-200 mesh, dichloromethane:methanol=50:1 to 10:1) to give the product (165.0 mg, yield: 79.7%).
(R)-3-methoxy-4-(4-((1-methylpiperidin-3-yl)amino)phthalazin-1-yl)benzonitrile (165.0 mg, 0.44 mmol, 1.0 eq.) was added to dichloromethane (4.0 mL), and the mixture was cooled to −20° C. in nitrogen atmosphere, followed by dropwise addition of boron tribromide (330.6 mg, 1.32 mmol, 3.0 eq.). The resulting mixture was gradually warmed to room temperature and reacted for 2 h. After the reaction was completed as monitored by TLC, methanol (5.0 mL) was added dropwise and slowly to the system to quench the reaction, and the mixture was concentrated under reduced pressure, followed by slow addition of a saturated aqueous sodium bicarbonate solution (50.0 mL). The resulting mixture was stirred for 30 min, and the pH was adjusted to 7-8, followed by extraction with dichloromethane (50.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by preparative thin-layer chromatography (dichloromethane:methanol=5:1) to give the product (60.0 mg, yield 37.9%).
1HNMR (300 MHz, DMSO-d6) δ (ppm): 10.55 (s, 1H), 8.49 (s, 1H), 7.79-7.92 (m, 2H), 7.37-7.50 (m, 5H), 4.64 (s, 1H), 3.12-3.16 (m, 2H), 2.63-2.68 (m, 4H), 1.75-2.01 (m, 5H).
Molecular formula: C21H21N5O Molecular weight: 359.43 LC-MS (Pos, m/z)=360.22[M+H]+.
1-Bromo-2-methoxy-4-methylbenzene (6.66 g, 30.10 mmol, 1.1 eq.) was dissolved in THE (50 mL), and the mixture was cooled to −50° C., followed by dropwise and slow addition of a solution of n-butyllithium in n-hexane (2.5 mol/L, 13.2 mL, 30.10 mmol, 1.1 eq.). The reaction solution was stirred for 30 min and then added dropwise and slowly to a solution of 4-fluoroisobenzofuran-1,3-dione (5 g, 30.10 mmol, 1.0 eq.) in THE (50 mL) at −60° C., and the resulting mixture was reacted at −60° C. in nitrogen atmosphere. After the completion of dropwise addition, the reaction solution was naturally warmed to room temperature, reacted for 0.5 h and then poured into a dilute hydrochloric acid solution (100 mL), followed by addition of EA (100 mL×2) for extraction. The organic phase was dried and concentrated to give a mixture (8.67 g, yield: 100%) of 3-fluoro-2-(2-methoxy-4-methylbenzoyl)benzoic acid and 2-fluoro-6-(2-methoxy-4-methylbenzoyl)benzoic acid.
The mixture (8.67 g, 30.10 mmol, 1.0 eq.) of 3-fluoro-2-(2-methoxy-4-methylbenzoyl)benzoic acid and 2-fluoro-6-(2-methoxy-4-methylbenzoyl)benzoic acid and 85% hydrazine hydrate (2.66 g, 45.15 mmol, 1.5 eq.) were dissolved in ethanol (100 mL), and the mixture was reacted at 80° C. for 3 h. After the generation of the target product as detected by LC-MS, the reaction solution was concentrated under reduced pressure, and the crude product was purified by column chromatography (PE:EA=3:1) to give a mixture (1.2 g, yield: 14.0%) of 5-fluoro-4-(2-methoxy-4-methylphenyl)phthalazin-1(2H)-one and 8-fluoro-4-(2-methoxy-4-methylphenyl)phthalazin-1(2H)-one.
5-Fluoro-4-(2-methoxy-4-methylphenyl)phthalazin-1(2H)-one, 8-fluoro-4-(2-methoxy-4-methylphenyl)phthalazin-1(2H)-one (1.2 g, 4.22 mmol, 1.0 eq.) and phosphorus oxychloride (1.29 g, 8.44 mmol, 2.0 eq.) were dissolved in acetonitrile (20 mL), and the mixture was reacted at 90° C. for 26 h. After the reaction was completed as monitored by TLC, the reaction solution was concentrated under reduced pressure, and the crude product was dissolved in EA (20 mL). Water (20 mL) was added, and the pH was adjusted to 9 with sodium carbonate, followed by liquid separation. The aqueous phase was extracted with EA (20 mL×2), and the organic phases were combined, dried and concentrated. The crude product was purified by silica gel column chromatography (PE:EA=6:1) to give a mixture (0.5 g, yield: 39.4%) of 1-chloro-5-fluoro-4-(2-methoxy-4-methylphenyl)phthalazine and 4-chloro-5-fluoro-1-(2-methoxy-4-methylphenyl)phthalazine.
The mixture (0.5 g, 1.65 mmol, 1.0 eq.) of 1-chloro-5-fluoro-4-(2-methoxy-4-methylphenyl)phthalazine and 4-chloro-5-fluoro-1-(2-methoxy-4-methylphenyl)phthalazine, tert-butyl (R)-3-aminopiperidine-1-carboxylate (396 mg, 1.98 mmol, 1.2 eq.), Pd2(dba)3 (146 mg, 0.16 mmol, 0.1 eq.), BINAP (205 mg, 0.33 mmol, 0.2 eq.) and cesium carbonate (1.08 g, 3.30 mmol, 2.0 eq.) were dissolved in 1,4-dioxane (10 mL), and the mixture was reacted for 22 h at 80° C. in nitrogen atmosphere. After the reaction was completed as detected by LC-MS, the reaction solution was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (MeOH:DCM=1:60) to give a mixture (400 mg, yield: 52.0%) of tert-butyl (R)-3-((5-fluoro-4-(2-methoxy-4-methylphenyl)phthalazin-1-yl)amino)piperidine-1-carboxylate and tert-butyl (R)-3-((8-fluoro-4-(2-methoxy-4-methylphenyl)phthalazin-1-yl)amino)piperidine-1-carboxylate.
The mixture (0.4 g, 0.85 mmol, 1.0 eq.) of tert-butyl (R)-3-((5-fluoro-4-(2-methoxy-4-methylphenyl)phthalazin-1-yl)amino)piperidine-1-carboxylate and tert-butyl (R)-3-((8-fluoro-4-(2-methoxy-4-methylphenyl)phthalazin-1-yl)amino)piperidine-1-carboxylate was dissolved in DCM (20 mL), and the mixture was cooled to −20° C., followed by dropwise addition of a 1 mol/L solution of boron tribromide in dichloromethane (2.6 mL, 2.55 mmol, 3.0 eq.). After the completion of dropwise addition, the reaction solution was naturally warmed to room temperature and reacted for 1 h. After the reaction was completed as detected by LC-MS, an appropriate amount of methanol was added into the flask to quench the reaction. The reaction solution was concentrated under reduced pressure, and the crude product was dissolved completely in DCM (10 mL), followed by addition of water (20 mL) and extraction with DCM (10 mL×2). The pH of the aqueous phase was adjusted to 9 with NaHCO3, followed by extraction with MeOH:DCM (1:10) (15 mL×2). The organic phase was dried and concentrated to give a mixture (80 mg, yield: 26.7%) of (R)-2-(8-fluoro-4-(piperidin-3-ylamino)phthalazin-1-yl)-5-methylphenol and (R)-2-(5-fluoro-4-(piperidin-3-ylamino)phthalazin-1-yl)-5-methylphenol.
The mixture (80 mg, 0.22 mmol, 1.0 eq.) of (R)-2-(8-fluoro-4-(piperidin-3-ylamino)phthalazin-1-yl)-5-methylphenol and (R)-2-(5-fluoro-4-(piperidin-3-ylamino)phthalazin-1-yl)-5-methylphenol was dissolved in methanol (2 mL), followed by addition of an aqueous formaldehyde solution (37%) (18 mg, 0.22 mmol, 1.0 eq.). The resulting mixture was stirred at room temperature for 5 min, followed by addition of sodium cyanoborohydride (16 mg, 0.22 mmol, 1.0 eq.). The mixture was reacted at room temperature for 5 min. After the generation of the target product as monitored by LC-MS, the reaction solution was concentrated under reduced pressure, and the crude product was purified by preparative thin-layer chromatography (DCM:MeOH=7:1) to give the product (compound 6) (R)-2-(8-fluoro-4-((1-methylpiperidin-3-yl)amino)phthalazin-1-yl)-5-methylphenol with a smaller Rf value (10 mg, yield: 12.5%).
1HNMR (400 MHz, DMSO-d6) δ (ppm): 9.26 (s, 1H), 8.24-8.22 (d, 1H), 7.90-7.85 (m, 1H), 7.58-7.54 (m, 1H), 7.27 (s, 1H), 7.15-7.13 (d, 1H), 6.70-6.69 (d, 1H), 4.52 (s, 1H), 2.97 (s, 1H), 2.51 (s, 3H), 2.50 (s, 2H), 2.30 (s, 3H), 2.02-2.00 (d, 3H), 1.86 (s, 1H), 1.71-1.69 (d, 1H).
Molecular formula: C21H23FN4O Molecular weight: 366.44 LC-MS (Pos, m/z)=367.20[M+H]+.
Simultaneously, the product (compound 9) (R)-2-(5-fluoro-4-((1-methylpiperidin-3-yl)amino)phthalazin-1-yl)-5-methylphenol with a larger Rf value (15 mg, yield: 18.8%) is also given.
1HNMR (400 MHz, DMSO-d6) δ (ppm): 9.51 (s, 1H), 7.82-7.76 (m, 1H), 7.70-7.65 (m, 1H), 7.33-7.31 (d, 1H), 7.15-7.13 (d, 1H), 6.81 (s, 1H), 6.78-6.76 (d, 1H), 6.69-6.65 (m, 1H), 4.56 (s, 1H), 2.95 (s, 1H), 2.52 (s, 1H), 2.50 (s, 3H), 2.42-2.40 (d, 2H), 2.33 (s, 3H), 1.80 (s, 3H), 1.66 (s, 1H).
The two-dimensional NOE spectrum showed coupling signals between 7.33-7.31 (d, 1H) and 7.15-7.13 (d, 1H) and coupling signals between 7.33-7.31 (d, 1H) and 9.51 (s, 1H).
Molecular formula: C21H23FN4O Molecular weight: 366.44 LC-MS (Pos, m/z)=367.22[M+H]+.
(R)-2-(6-bromo-4-((1-methylpiperidin-3-yl)amino)phthalazin-1-yl)-5-methylphenol (100 mg, 0.23 mmol, 1.0 eq.), 2-di-tert-butylphosphino-2′,4′,6′-triisopropylbiphenyl (21 mg, 0.05 mmol, 0.2 eq.), cesium carbonate (150 mg, 0.46 mmol, 2.0 eq.) and allylpalladium chloride dimer (18 mg, 0.05 mmol, 0.2 eq.) were added to a mixed solution of MeOH (1 mL) and dimethylbenzene (2 mL), and the mixture was reacted at 80° C. for 1 h in nitrogen atmosphere. After the reaction was completed as monitored by LC-MS, the reaction solution was concentrated under reduced pressure, and the crude product was purified by preparative thin-layer chromatography (MeOH:DCM=1:7) to give the product (15 mg, yield: 17.2%).
1HNMR (300 MHz, DMSO-d6) δ (ppm): 7.74 (s, 1H), 7.50-7.47 (d, 1H), 7.40-7.37 (d, 1H), 7.17-7.15 (d, 1H), 6.80-6.75 (d, 2H), 4.48 (s, 1H), 3.96 (s, 3H), 3.42 (s, 3H), 2.88 (s, 1H), 2.38 (s, 3H), 2.33 (s, 4H), 2.19-2.02 (d, 2H), 1.70-1.54 (d, 2H).
Molecular formula: C22H26N4O2 Molecular weight: 378.48 LC-MS (Pos, m/z): 379.17[M+H]+.
(R)-2-(6-bromo-4-((1-methylpiperidin-3-yl)amino)phthalazin-1-yl)-5-methylphenol (120 mg, 0.28 mmol, 1.0 eq.), zinc cyanide (33 mg, 0.28 mmol, 1.0 eq.) and tetrakis(triphenylphosphine)palladium (65 mg, 0.026 mmol, 0.2 eq.) were dissolved in DMAC (2 mL), and the mixture was reacted at 120° C. for 16 h in nitrogen atmosphere. After the reaction was completed as monitored by LC-MS, the reaction solution was cooled to room temperature and then poured into water (10 mL), followed by extraction with EA (10 mL×3). The organic phases were combined, washed with water (15 mL×3) and concentrated under reduced pressure, and the crude product was purified by preparative thin-layer chromatography (MeOH:DCM=1:8) to give the product (50 mg, yield: 48.1%).
1HNMR (300 MHz, DMSO-d6) δ (ppm): 9.61 (s, 1H), 9.08 (s, 1H), 8.13-8.11 (d, 1H), 7.61-7.58 (d, 1H), 7.50-7.52 (d, 1H), 7.19-7.17 (d, 1H), 6.83-6.78 (t, 2H), 4.53 (s, 1H), 3.41 (s, 4H), 2.96-2.92 (d, 1H), 2.45 (s, 2H), 2.34 (s, 3H), 2.02-1.88 (d, 2H), 1.73-1.59 (m, 2H).
Molecular formula: C22H23N5O Molecular weight: 373.46 LC-MS (Pos, m/z): 374.17[M+H]+.
Tert-butyl (R)-3-((7-bromo-4-(2-methoxy-4-methylphenyl)phthalazin-1-yl)amino)piperidine-1-carboxylate (0.5 g, 0.94 mmol, 1.0 eq.), cyclopropylboronic acid (242 mg, 2.82 mmol, 3.0 eq.), tricyclohexylphosphorus (131 mg, 0.47 mmol, 0.5 eq.), palladium acetate (43 mg, 0.19 mmol, 0.2 eq.) and potassium phosphate (399 mg, 1.88 mmol, 2.0 eq.) were added to a mixed solution of toluene (10 mL) and water (2 mL), and the mixture was reacted at 100° C. for 2 h in nitrogen atmosphere. After the reaction was completed as detected by TLC, the reaction solution was poured into water (10 mL), and EA (10 mL×3) was added for extraction. The organic phase was dried and concentrated to give the product (400 mg, yield: 87.1%).
Tert-butyl (R)-3-((7-cyclopropyl-4-(2-methoxy-4-methylphenyl)phthalazin-1-yl)amino)piperidine-1-carboxylate (400 mg, 0.81 mmol, 1.0 eq.) was dissolved in DCM (10 mL), followed by dropwise addition of a 1 mol/L solution of boron tribromide in dichloromethane (2.4 mL, 2.43 mmol, 3.0 eq.) at room temperature. The mixture was reacted at room temperature for 20 min. After the reaction was completed as detected by LC-MS, an appropriate amount of methanol was added to the system to quench the reaction. The reaction solution was concentrated under reduced pressure, and the crude product was dissolved in water (10 mL). The aqueous phase was extracted with DCM (10 mL×2), and the aqueous phase was retained. The pH was adjusted to 9 with NaHCO3, followed by extraction with DCM (10 mL×4). The organic phases were combined, dried and concentrated to give the product (150 mg, yield: 47.7%).
(R)-7-cyclopropyl-4-(2-methoxy-4-methylphenyl)-N-(piperidin-3-yl) phthalazin-1-amine (150 mg, 0.40 mmol, 1.0 eq.) was dissolved in methanol (3 mL), followed by addition of an aqueous formaldehyde solution (37%) (32 mg, 0.40 mmol, 1.0 eq.). The resulting mixture was stirred at room temperature for 5 min, followed by addition of sodium cyanoborohydride (25 mg, 0.40 mmol, 1.0 eq.). The mixture was reacted at room temperature for 5 min. After the reaction was completed as monitored by LC-MS, the reaction solution was concentrated under reduced pressure, and water (5 mL) and DCM (10 mL) were added to the system, followed by liquid separation. The aqueous phase was extracted with DCM (10 mL×2), and the organic phases were combined, dried and concentrated. The crude product was purified by preparative thin-layer chromatography (DCM:MeOH=7:1) to give the product (45 mg, yield: 29.0%).
1HNMR (300 MHz, DMSO-d6) δ (ppm): 8.03 (s, 1H), 7.54-7.51 (d, 1H), 7.43-7.41 (d, 1H), 7.16-7.14 (d, 1H), 6.80-6.75 (t, 2H), 4.54 (s, 1H), 3.43 (s, 5H), 2.99 (s, 1H), 2.51 (s, 4H), 2.33 (s, 3H), 2.16-2.10 (m, 1H), 2.02-1.88 (t, 2H), 1.74-1.62 (m, 2H), 1.13-1.10 (t, 2H), 0.92-0.91 (d, 2H).
Molecular formula: C24H28N4O Molecular weight: 388.52 LC-MS (Pos, m/z): 389.23[M+H]+.
4,7-Dichlorothieno[2,3-d]pyridazine (1.0 g, 4.88 mmol, 1.0 eq.) and tert-butyl (R)-3-aminopiperidine-1-carboxylate (1.17 g, 5.86 mmol, 1.2 eq.) were dissolved in DMA (10 mL), and the mixture was heated to 120° C. and reacted for 20 h in nitrogen atmosphere. Water (50 mL) was added to quench the reaction, and EA (50 mL) was added for extraction. The organic phase was washed with water (30 mL×3), dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (DCM:MeOH=100:1 to 50:1) to give the product (1.40 g, yield: 77.8%).
(2-Methoxy-4-methylphenyl)boronic acid (0.630 g, 3.80 mmol, 1.0 eq.), tert-butyl (R)-3-((7-chlorothieno[2,3-d]pyridazin-4-yl)amino)piperidine-1-carboxylate (1.40 g, 3.80 mmol, 1.0 eq.), Pd(dppf)Cl2 (0.139 g, 0.190 mmol, 0.05 eq.) and NaHCO3 (0.638 g, 7.60 mmol, 2.0 eq.) were added successively to 1,4-dioxane (20 mL), followed by addition of H2O (10 mL). The mixture was heated to 110° C. and reacted for 6 h in nitrogen atmosphere. Water (60 mL) was added, and EA (50 mL) was added for extraction. The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (DCM:MeOH=100:1 to 20:1) to give the product (920 mg, yield: 53.3%).
Tert-butyl (R)-3-((7-(2-methoxy-4-methylphenyl)thieno[2,3-d]pyridazin-4-yl)amino)piperidine-1-carboxylate (900 mg, 1.98 mmol, 1.0 eq.) was dissolved in DCM (18 mL), and the mixture was cooled to −10° C., followed by dropwise addition of BBr3 (1.98 g, 7.92 mmol, 4.0 eq.). The mixture was reacted for 4 h. MeOH (5 mL) was added to quench the reaction, a saturated aqueous NaHCO3 solution was added to adjust the pH to 8, and DCM (30 mL×2) was added for extraction. The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (DCM:MeOH=50:1 to 10:1) to give the product (490 mg, yield: 72.7%).
(R)-5-methyl-2-(4-(piperidin-3-ylamino)thieno[2,3-d]pyridazin-7-yl)phenol (490 mg, 1.44 mmol, 1.0 eq.) was dissolved in MeOH (4 mL), followed by addition of an aqueous formaldehyde solution (37%) (351 mg, 4.32 mmol, 3.0 eq.). The mixture was stirred at room temperature for 30 min, followed by addition of NaBH3CN (271 mg, 4.32 mmol, 3.0 eq.). The resulting mixture was reacted for 1 h. A saturated aqueous NaHCO3 solution was added to quench the reaction, and DCM (30 mL×2) was added for extraction. The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was separated by preparative thin-layer chromatography (DCM:MeOH=15:1) to give the product (120 mg, yield: 23.5%).
1HNMR (400 MHz, DMSO-d6) δ (ppm): 12.26 (s, 1H), 8.25 (d, J=5.2 Hz, 1H), 8.07 (d, J=5.2 Hz, 1H), 7.78 (d, J=8.4 Hz, 1H), 7.32 (d, J=6.8 Hz, 1H), 6.84 (d, J=6.4 Hz, 2H), 4.40 (s, 1H), 3.16 (d, J=9.6 Hz, 1H), 2.81 (d, J=10.3 Hz, 1H), 2.32 (s, 6H), 2.09-1.99 (m, 3H), 1.82-1.79 (m, 1H), 1.69-1.60 (m, 1H), 1.50-1.45 (m, 1H).
Molecular formula: C19H22N4OS Molecular weight: 354.47 LC-MS (Pos, m/z)=355.13[M+H]+.
1-Bromo-2-methoxy-4-methylbenzene (14.16 g, 70.41 mmol, 1.05 eq.) was dissolved in anhydrous THE (100 mL), followed by dropwise addition of n-BuLi (32.2 mL, 80.47 mmol, 1.2 eq.) at −78° C. After the completion of dropwise addition, the mixture was reacted for 30 min. At −78° C., the reaction solution was added dropwise to a solution of 3,4-pyridine dicarboxylic anhydride (10 g, 67.06 mmol, 1.0 eq.) in THE (100 mL). After the completion of dropwise addition, the mixture was reacted at room temperature for 1 h. After the reaction was completed as detected by TLC, the reaction solution was poured into a mixed solution of ice water (50 mL) and 2 mol/L hydrochloric acid (50 mL), followed by extraction with dichloromethane (200 mL×3). The organic phases were combined, dried and concentrated to give the crude product (20 g). 5 g of the crude product was subjected to silica gel column chromatography (petroleum ether:ethyl acetate=1:3 (containing 0.5% AcOH), dichloromethane:methanol=10:1 (containing 0.5% AcOH)) to give the product (2.5 g, yield: 55.6%).
4-(2-Methoxy-4-methylbenzoyl)nicotinic acid (2.5 g, 9.22 mmol, 1.0 eq.) was dissolved in DMF (30 mL), followed by addition of potassium carbonate (1.91 g, 13.83 mmol, 1.5 eq.) and iodomethane (1.31 g, 9.22 mmol, 1.0 eq.). The mixture was reacted at room temperature for 2 h. After the reaction was completed as detected by TLC, the reaction solution was poured into water (20 mL), followed by extraction with EA (50 mL×3). The organic phases were combined, dried over anhydrous sodium sulfate and concentrated to give the product (1.1 g, yield: 41.8%).
Methyl 4-(2-methoxy-4-methylbenzoyl)nicotinate (1.1 g, 3.86 mmol, 1.0 eq.) was dissolved in ethanol (10 mL), followed by addition of 85% hydrazine hydrate (341 mg, 5.79 mmol, 1.5 eq.). The mixture was heated to 85° C. and reacted for 10 min. After the reaction was completed as detected by LC-MS, the reaction solution was cooled to room temperature, and solids precipitated. The mixture was filtered to give the product (640 mg, yield: 62.1%).
Acetonitrile (8 mL) was added to 1-(2-methoxy-4-methylphenyl)pyrido[3,4-d]pyridazin-4-ol (640 mg, 2.39 mmol, 1.0 eq.), followed by addition of phosphorus oxychloride (733 mg, 4.78 mmol, 2.0 eq.). The mixture was reacted at 90° C. for 16 h. After the reaction was completed as detected by TLC, the reaction solution was concentrated, and the residue was poured into ice water (10 mL), followed by extraction with dichloromethane (20 mL). The organic phase was dried and concentrated, and the crude product was subjected to silica gel column chromatography (petroleum ether:ethyl acetate=3:1) to give the product (536 mg, yield: 78.4%).
4-Chloro-1-(2-methoxy-4-methylphenyl)pyrido[3,4-d]pyridazine (536 mg, 1.876 mmol, 1.0 eq.) was dissolved in DMA (5 mL), followed by addition of (R)-1-tert-butoxycarbonyl-3-aminopiperidine (752 mg, 3.752 mmol, 2.0 eq.). The mixture was refluxed at 120° C. for 16 h. After the reaction was completed as detected by TLC, the reaction solution was added dropwise into water (5 mL), ethyl acetate (10 mL) was added for extraction, and DCM (10 mL×3) was added for extraction. The organic phases were combined, dried and concentrated to give the product (647 mg, yield: 76.7%).
Tert-butyl (R)-3-((1-(2-methoxy-4-methylphenyl)pyrido[3,4-d]pyridazin-4-yl)amino)piperidine-1-carboxylate (647 mg, 1.44 mmol, 1.0 eq.) was dissolved in DCM (5 mL), followed by dropwise addition of boron tribromide (1.08 g, 4.32 mmol, 3.0 eq.) at −20° C. The mixture was reacted at room temperature for 1 h. After there remained some of the materials as detected by LC-MS, the reaction solution was supplemented with boron tribromide (1.08 g, 4.32 mmol, 3.0 eq.). The resulting mixture was reacted at room temperature for 1 h. After the reaction was completed as detected by LC-MS, methanol (3 mL) and water (10 mL) were added dropwise and slowly to the reaction solution under an ice bath, followed by back-extraction with DCM (10 mL). The pH of the aqueous phase was adjusted to be neutral with a saturated aqueous sodium bicarbonate solution, followed by back-extraction with DCM (10 mL). The aqueous phase was concentrated, dissolved in a mixed solvent of DCM/MeOH, dried over anhydrous magnesium sulfate and concentrated, and the crude product was directly used in the next step.
(R)-5-methyl-2-(4-((piperidin-3-yl)amino)pyrido[3,4-d]pyridazin-1-yl)phenol (1.44 mmol, 1.0 eq.) was dissolved in methanol (5 mL), followed by addition of an aqueous acetaldehyde solution (38.5%) (112 mg, 1.44 mmol, 1.0 eq.). The mixture was stirred at room temperature for 5 min, followed by addition of sodium cyanoborohydride (100 mg, 1.584 mmol, 1.1 eq.). The resulting mixture was reacted at room temperature for 5 min. After the reaction was completed as detected by TLC, the reaction solution was concentrated, and the crude product was purified by preparative thin-layer chromatography (dichloromethane:methanol=7:1) to give the product (12 mg, two-step yield: 2.3%).
1H-NMR (300 MHz, DMSO-d6) δ (ppm): 9.86 (brs, 1H), 9.67 (s, 1H), 8.86-8.88 (d, 1H), 7.84 (s, 1H), 7.33-7.35 (d, 1H), 7.19-7.21 (d, 1H), 6.79-6.84 (m, 2H), 4.66 (s, 1H), 3.13 (s, 3H), 2.64-2.74 (m, 4H), 2.34 (s, 3H), 1.77-2.02 (m, 4H).
Molecular formula: C20H23N5O Molecular weight: 349.44 LC-MS=350.23[M+H]+
4-Bromo-1-iodo-2-methoxybenzene (10 g, 31.96 mmol, 1.0 eq.) was dissolved in anhydrous THE (100 mL), followed by dropwise addition of a solution of i-PrMgCl LiCl in THE (1.3 mol/L, 26 mL, 33.6 mmol, 1.05 eq.) at −20° C. After the completion of dropwise addition, the mixture was reacted for 30 min. At −20° C., the above reaction solution was added dropwise to a solution of 2,3-pyridine dicarboxylic anhydride (4.77 g, 31.96 mmol, 1.0 eq.) in THE (100 mL). After the completion of dropwise addition, the mixture was reacted at room temperature for 16 h. After the reaction was completed as detected by TLC, the reaction solution was poured into a saturated aqueous ammonium chloride solution (200 mL), followed by concentration and extraction with dichloromethane (200 mL×5). The organic phases were combined, dried over anhydrous magnesium sulfate and concentrated, and the crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=2:1, v/v, with 5% acetic acid added) to give the product (3.9 g, yield: 36.3%).
3-(4-Bromo-2-methoxybenzoyl)nicotinic acid (3.9 g, 11.6 mmol, 1.0 eq.) was added to EtOH (40 mL), followed by addition of 85% hydrazine hydrate (1.37 g, 23.2 mmol, 2.0 eq.). The mixture was heated to 90° C. and reacted for 8 h. After the reaction was completed as detected by TLC, the reaction solution was concentrated and filtered, and the filter cake was washed with ethanol to give the product (2.02 g, yield: 52.4%).
5-(4-Bromo-2-methoxyphenyl)pyrido[2,3-d]pyridazin-8-ol (2.02 g, 6.08 mmol, 1.0 eq.) was added to acetonitrile (20 mL), followed by addition of phosphorus oxychloride (1.86 g, 12.16 mmol, 2.0 eq.). The mixture was reacted at 100° C. for 16 h. After the reaction was completed as detected by TLC, the reaction solution was concentrated, and the concentrated solution was poured into ice water (10 mL). The pH was adjusted to alkalinity with sodium bicarbonate, followed by extraction with dichloromethane (30 mL×3). The organic phases were combined, dried and concentrated, and the crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=5:1, v/v) to give the product (2.0 g, yield: 93.8%).
5-(4-Bromo-2-methoxyphenyl)-8-chloropyrido[2,3-d]pyridazine (2 g, 5.7 mmol, 1.0 eq.) was dissolved in dichloromethane (20 mL), followed by dropwise and slow addition of a solution of boron tribromide in n-heptane (1 mol/L, 8.6 mL, 8.6 mmol, 1.5 eq.) at −20° C. After the completion of dropwise addition, the mixture was reacted at room temperature for 10 min. After the reaction was completed as detected by TLC, a saturated aqueous sodium bicarbonate solution (10 mL) was added dropwise and slowly to the reaction solution under an ice bath, followed by extraction with DCM (20 mL×3). The organic phases were combined, dried and concentrated, and the crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=2:1, v/v) to give the product (900 mg, yield: 47.3%).
5-Bromo-2-(8-chloropyrido[2,3-d]pyridazin-5-yl)phenol (900 mg, 2.7 mmol, 1.0 eq.) was dissolved in DMAc (10 mL), followed by addition of (R)-1-tert-butoxycarbonyl-3-aminopiperidine (1.08 g, 5.4 mmol, 2.0 eq.). The mixture was refluxed at 120° C. for 1 h. After the reaction was completed as detected by TLC, the reaction solution was added dropwise into water (5 mL), followed by extraction with ethyl acetate (10 mL×2). The organic phases were combined, dried and concentrated, and the crude product was purified by silica gel column chromatography (dichloromethane:methanol=200:1) to give the product (1.05 g, yield: 78.3%). (1900326-179)
Tert-butyl (R)-3-((5-(4-bromo-2-methoxyphenyl)pyrido[2,3-d]pyridazin-8-yl)amino)piperidine-1-carboxylate (500 mg, 1 mmol, 1.0 eq.) was dissolved in DMAc (5 mL), followed by addition of zinc cyanide (94 mg, 0.8 mmol, 0.8 eq.) and tetrakis(triphenylphosphine)palladium (112 mg, 0.1 mmol, 0.1 eq.) in nitrogen atmosphere. The mixture was reacted at 120° C. for 45 min. After the reaction was completed as detected by LC-MS, the reaction solution was poured into water, followed by extraction with EA (10 mL×2). The organic phases were combined, dried and concentrated, and the crude product was purified by silica gel column chromatography (dichloromethane:methanol=200:1 to 100:1, v/v) to give the product (480 mg, crude product).
Tert-butyl (R)-3-((5-(4-cyano-2-hydroxyphenyl)pyrido[2,3-d]pyridazin-8-yl)amino)piperidine-1-carboxylate (1 mmol, 1.0 eq.) was dissolved in dichloromethane (2 mL), and the solution was added dropwise into a 4 mol/L solution of hydrogen chloride in 1,4-dioxane (4 mL). After the completion of dropwise addition, a large number of solids precipitated. The mixture was reacted at room temperature for 5 min. After the reaction was completed as detected by TLC, water (5 mL) was added to the reaction solution, the pH was adjusted to alkalinity with sodium bicarbonate, and the aqueous phase was concentrated to give the crude product. The crude product was dissolved in dichloromethane/methanol (20:1, 20 mL), and the solution was filtered. The filtrate was dried and concentrated to give the product (279 mg, two-step yield: 80.6%).
(R)-3-hydroxy-4-(8-(piperidin-3-ylamino)pyrido[2,3-d]pyridazin-5-yl)benzonitrile (279 mg, 0.832 mmol, 1.0 eq.) was dissolved in methanol (3 mL), followed by addition of an aqueous formaldehyde solution (37%) (68 mg, 0.832 mmol, 1.0 eq.). The mixture was stirred at room temperature for 5 min, followed by addition of sodium cyanoborohydride (58 mg, 0.915 mmol, 1.1 eq.). The resulting mixture was reacted at room temperature for 5 min. After the reaction was completed as detected by TLC, the reaction solution was concentrated, saturated brine (1 mL) was added, and the pH was adjusted to alkalinity with sodium bicarbonate, followed by extraction with a mixed solvent (dichloromethane:methanol:ammonia solution=20:1:0.5, 20 mL×3). The organic phases were combined, dried and concentrated, and the crude product was purified by silica gel column chromatography (dichloromethane:methanol=50:1 to 20:1, v/v) and lyophilized to give the product (112 mg, yield: 37.3%).
1H-NMR (400 MHz, DMSO-d6) δ (ppm): 11.93 (brs, 1H), 9.14 (s, 1H), 8.92-8.90 (d, 1H), 8.20-8.18 (d, 1H), 7.93 (s, 1H), 7.61-7.60 (d, 1H), 7.38-7.31 (m, 2H), 4.43 (s, 1H), 3.11-3.09 (d, 1H), 2.75-2.73 (d, 1H), 2.24 (s, 3H), 1.99-1.76 (m, 4H), 1.63-1.45 (m, 2H).
Molecular formula: C20H20N6O Precise molecular weight: 360.17 LC-MS (m/z)=361.11 [M+H]+
2-Methoxy-4-methylbenzoic acid (5.0 g, 30.08 mmol, 1.0 eq.) and DMF (0.1 mL) were dissolved in DCM (100 mL), followed by addition of oxalyl chloride (2.30 g, 18.05 mmol, 0.6 eq.). The mixture was reacted at room temperature for 15 h, and the reaction solution was concentrated under reduced pressure to give the product (5.55 g, yield: 100%).
Methyl 5-bromo-2-iodobenzoate (10.3 g, 30.06 mmol, 1.0 eq.) was dissolved in THE (50 mL), and the mixture was cooled to −30° C., followed by dropwise and slow addition of a solution of an isopropylmagnesium chloride lithium chloride complex in tetrahydrofuran (1.3 mol/L, 25.4 mL, 33.07 mmol, 1.1 eq.). The resulting mixture was stirred for 30 min. The above reaction solution was added dropwise and slowly to a solution of 2-methoxy-4-methylbenzoyl chloride (5.55 g, 30.06 mmol, 1.0 eq.) in THE (50 mL) at −60° C., and the mixture was reacted at −70° C. After the completion of dropwise addition, the mixture was naturally warmed to room temperature and reacted for 2.5 h. The reaction solution was poured into water (100 mL), followed by extraction with EA (100 mL×2). The organic phase was dried and concentrated, and the crude product was purified by silica gel column chromatography (PE:EA=20:1) to give the product (3.5 g, yield: 32.1%).
Methyl 5-bromo-2-(2-methoxy-4-methylbenzoyl)benzoate (3.5 g, 9.63 mmol, 1.0 eq.) and 85% hydrazine hydrate (0.85 g, 14.44 mmol, 1.5 eq.) were dissolved in ethanol (35 mL), and the mixture was reacted at 80° C. for 1.5 h. After the reaction was completed as monitored by TLC, the reaction solution was cooled to room temperature, and a large number of solids precipitated. The mixture was filtered under vacuum to give the product (1.8 g, yield: 54.2%).
7-Bromo-4-(2-methoxy-4-methylphenyl)phthalazin-1-ol (1.8 g, 5.21 mmol, 1.0 eq.) and phosphorus oxychloride (1.6 g, 10.42 mmol, 2.0 eq.) were dissolved in acetonitrile (20 mL), and the mixture was reacted at 90° C. for 2 h. After the reaction was completed as monitored by LC-MS, the reaction solution was concentrated under reduced pressure, and the crude product was dissolved in EA (30 mL), followed by addition of water (10 mL). The pH was adjusted to 9 with a saturated aqueous sodium bicarbonate solution, followed by liquid separation. The aqueous phase was extracted with EA (20 mL), and the organic phases were combined, dried and concentrated to give the product (1.6 g, yield: 84.6%).
6-Bromo-4-chloro-1-(2-methoxy-4-methylphenyl)phthalazine (1.6 g, 4.40 mmol, 1.0 eq.) and tert-butyl (R)-3-aminopiperidine-1-carboxylate (1.76 g, 8.80 mmol, 2.0 eq.) were dissolved in DMAC (16 mL), and the mixture was reacted at 120° C. for 26 h. After the reaction was completed as detected by LC-MS, the reaction solution was poured into water (50 mL), and solids precipitated. The mixture was filtered under vacuum, and the filter cake was dissolved in EA (50 mL) and dried. The organic phase was concentrated under reduced pressure to give the product (1.2 g, yield: 51.7%).
Tert-butyl (R)-3-((7-bromo-4-(2-methoxy-4-methylphenyl)phthalazin-1-yl)amino)piperidine-1-carboxylate (1.0 g, 1.89 mmol, 1.0 eq.) was dissolved in DCM (10 mL), and the mixture was cooled to −20° C., followed by dropwise addition of a 1 mol/L solution of boron tribromide in dichloromethane (5.7 mL, 5.67 mmol, 3.0 eq.). After the completion of dropwise addition, the resulting mixture was naturally warmed to room temperature and reacted for 1 h. After the reaction was completed as detected by LC-MS, an appropriate amount of methanol was added to the flask to quench the reaction. The reaction solution was concentrated under reduced pressure, and the crude product was dissolved in water (20 mL). EA (20 mL×2) was added for extraction, and the aqueous phase was retained. The pH was adjusted to 9 with NaHCO3, followed by extraction with DCM (20 mL×2). The organic phases were combined, dried and concentrated to give the product (400 mg, yield: 51.2%).
(R)-2-(6-bromo-4-(piperidin-3-ylamino)phthalazin-1-yl)-5-methylphenol (400 mg, 0.96 mmol, 1.0 eq.) was dissolved in methanol (4 mL), followed by addition of an aqueous formaldehyde solution (37%) (78 mg, 0.96 mmol, 1.0 eq.). The mixture was stirred at room temperature for 5 min, followed by addition of sodium cyanoborohydride (60 mg, 0.96 mmol, 1.0 eq.). The mixture was reacted at room temperature for 5 min. After the generation of the target product as monitored by LC-MS, the reaction solution was concentrated under reduced pressure, and the crude product was slurried with water (10 mL) and filtered under vacuum. The filter cake was purified by silica gel column chromatography (DCM:MeOH=30:1) to give the product (200 mg, yield: 48.8%).
1HNMR (300 MHz, DMSO-d6) δ (ppm): 9.60 (s, 1H), 8.75 (s, 1H), 7.97-7.94 (d, 1H), 7.45-7.42 (d, 1H), 7.35 (s, 1H), 7.17-7.15 (d, 1H), 6.82-6.77 (t, 2H), 4.53 (s, 1H), 3.03 (s, 2H), 2.51 (s, 4H), 2.33 (s, 4H), 2.02-1.92 (d, 2H), 1.74-1.62 (m, 2H).
Molecular formula: C21H23BrN4O Molecular weight: 427.35 LC-MS (Pos, m/z): 427.05/429.08[M+H]+.
(2-Methoxy-4-methylphenyl)boronic acid (0.809 g, 4.88 mmol, 1.0 eq.), 4,7-dichlorothieno[2,3-d]pyridazine (1.00 g, 4.88 mmol, 1.0 eq.), Pd(dppf)Cl2 (0.179 g, 0.244 mmol, 0.05 eq.) and NaHCO3 (0.820 g, 9.76 mmol, 2.0 eq.) were added successively to 1,4-dioxane (20 mL), followed by addition of H2O (10 mL). The mixture was heated to 110° C. and reacted for 2 h in nitrogen atmosphere. Water (60 mL) was added, and EA (50 mL) was added for extraction. The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (PE:EA=5:1 to 1:1) to give the product (820 mg, yield: 57.9%).
7-Chloro-4-(2-methoxy-4-methylphenyl)thieno[2,3-d]pyridazine (820 mg, 2.92 mmol, 1.0 eq.) and tert-butyl (R)-3-aminopiperidine-1-carboxylate (877 mg, 4.38 mmol, 1.5 eq.) were dissolved in DMA (10 mL), and the mixture was heated to 120° C. and reacted for 16 h in nitrogen atmosphere. Water (50 mL) was added to quench the reaction, and EA (50 mL) was added for extraction. The organic phase was washed with water (20 mL×3), dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (DCM:MeOH=100:1 to 50:1) to give the product (610 mg, yield: 47.6%).
Tert-butyl (R)-3-((4-(2-methoxy-4-methylphenyl)thieno[2,3-d]pyridazin-7-yl)amino)piperidine-1-carboxylate (610 mg, 1.34 mmol, 1.0 eq.) was dissolved in DCM (12 mL), and the mixture was cooled to −10° C., followed by dropwise addition of BBr3 (1.34 g, 5.36 mmol, 4.0 eq.). The mixture was reacted for 4 h. MeOH (5 mL) was added to quench the reaction, a saturated aqueous NaHCO3 solution was added to adjust the pH to 8, and DCM (30 mL×2) was added for extraction. The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to give the product (380 mg, yield: 83.2%).
(R)-5-methyl-2-(7-(piperidin-3-ylamino)thieno[2,3-d]pyridazin-4-yl)phenol (380 mg, 1.12 mmol, 1.0 eq.) was dissolved in MeOH (4 mL), followed by addition of an aqueous formaldehyde solution (37%) (273 mg, 3.36 mmol, 3.0 eq.). The mixture was stirred at room temperature for 30 min. NaBH3CN (211 mg, 3.36 mmol, 3.0 eq.) was then added and the mixture was reacted for 1 h. A saturated aqueous NaHCO3 solution was added to quench the reaction, and DCM (30 mL×2) was added for extraction. The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was separated by preparative thin-layer chromatography (DCM:MeOH=15:1) to give the product (70 mg, yield: 17.7%).
1HNMR (400 MHz, DMSO-d6) δ (ppm): 11.43 (s, 1H), 8.12 (d, J=5.2 Hz, 1H), 7.56 (d, J=5.6 Hz, 1H), 7.52 (d, J=8.0 Hz, 1H), 7.07 (d, J=7.6 Hz, 1H), 6.82 (s, 1H), 6.79 (d, J=8.0 Hz, 1H), 4.40-4.38 (m, 1H), 3.09 (d, J=8.8 Hz, 1H), 2.75 (d, J=10.4 Hz, 1H), 2.32 (s, 3H), 2.26 (s, 3H), 1.99-1.96 (m, 3H), 1.77-1.74 (m, 1H), 1.66-1.57 (m, 1H), 1.50-1.41 (m, 1H).
Molecular formula: C19H22N4OS Molecular weight: 354.47 LC-MS (Pos, m/z)=355.13[M+H]+.
4-Bromo-1-iodo-2-methoxybenzene (5.0 g, 15.97 mmol, 1.0 eq.) was added to tetrahydrofuran (50.0 mL), and the mixture was cooled to −20° C. in nitrogen atmosphere, followed by dropwise addition of a solution of isopropylmagnesium chloride-lithium chloride in tetrahydrofuran (1.3 mol/L, 14.7 mL, 19.17 mmol, 1.2 eq.). The resulting mixture was reacted at −20° C. for 0.5 h, and then the reaction solution was added dropwise to a solution of phthalic anhydride (2.6 g, 17.56 mmol, 1.1 eq.) in tetrahydrofuran (20.0 mL) cooled to −20° C. The mixture was warmed to room temperature and reacted for 2 h. After the reaction was completed as monitored by TLC, ethyl acetate (200.0 mL) was added, followed by washing with a saturated aqueous ammonium chloride solution (100.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to give the product (5.0 g, yield: 93.4%).
2-(4-Bromo-2-methoxybenzoyl)benzoic acid (5.0 g, 14.91 mmol, 1.0 eq.), potassium carbonate (3.0 g, 22.36 mmol, 1.5 eq.) and iodomethane (2.7 g, 19.39 mmol, 1.3 eq.) were added to N,N-dimethylformamide (30.0 mL), and the mixture was reacted at room temperature for 4 h. After the reaction was completed as monitored by TLC, ethyl acetate (200.0 mL) was added, followed by washing with water (100.0 mL×3). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (specification of silica gel: 100-200 mesh, petroleum ether:ethyl acetate=30:1 to 10:1) to give the product (3.68 g, yield: 70.7%).
Methyl 2-(4-bromo-2-methoxybenzoyl)benzoate (2.0 g, 5.72 mmol, 1.0 eq.) was added to ethanol (20.0 mL) and hydrazine hydrate (85%) (505.9 mg, 8.59 mmol, 1.5 eq.), and the mixture was reacted at room temperature for 1 h. After the reaction was completed as monitored by LC-MS, the reaction solution was filtered, and the filter cake was washed with ethanol (50.0 mL) and dried to give the product (1.25 g, yield: 66.1%).
4-(4-Bromo-2-methoxyphenyl)phthalazin-1-ol (1.25 g, 3.77 mmol, 1.0 eq.) and phosphorus oxychloride (3.47 g, 22.64 mmol, 6.0 eq.) were added to acetonitrile (20.0 mL), and the mixture was refluxed for 1 h. After the reaction was completed as monitored by TLC, the reaction solution was poured into ice water (200.0 mL), and ethyl acetate (100.0 mL) was added for extraction. The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to give the product (1.1 g, yield: 83.9%).
1-(4-Bromo-2-methoxyphenyl)-4-chlorophthalazine (1.1 g, 3.14 mmol, 1.0 eq.) and tert-butyl (R)-3-aminopiperidine-1-carboxylate (1.26 g, 6.29 mmol, 2.0 eq.) were added to N,N-dimethylacetamide (20.0 mL), and the mixture was stirred at 120° C. for 12 h. After the reaction was completed as monitored by TLC, the reaction solution was cooled to room temperature, and ethyl acetate (100.0 mL) was added, followed by washing with water (100.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (specification of silica gel: 100-200 mesh, petroleum ether:ethyl acetate=4:1 to 1:1) to give the product (920.0 mg, yield: 57.1%).
Tert-butyl (R)-3-((4-(4-bromo-2-methoxyphenyl)phthalazin-1-yl)amino)piperidine-1-carboxylate (920.0 mg, 1.79 mmol, 1.0 eq.) was added to dichloromethane (6.0 mL) and trifluoroacetic acid (6.0 mL), and the mixture was stirred at room temperature for 1 h. After the reaction was completed as monitored by TLC, the pH was adjusted to 7-8 with a saturated aqueous sodium bicarbonate solution, followed by extraction with dichloromethane (100.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to give the product (720 mg, yield: 97.3%).
(R)-4-(4-bromo-2-methoxyphenyl)-N-(piperidin-3-yl) phthalazin-1-amine (720.0 mg, 1.74 mmol, 1.0 eq.) and an aqueous formaldehyde solution (37%) (169.6 mg, 2.09 mmol, 1.2 eq.) were added to methanol (8.0 mL), and the mixture was stirred at room temperature for 0.5 h, followed by addition of sodium cyanoborohydride (153.0 mg, 2.43 mmol, 1.4 eq.). The resulting mixture was reacted at room temperature for 1 h. After the reaction was completed as monitored by TLC, a saturated aqueous sodium bicarbonate solution (50.0 mL) was added, and the mixture was stirred for 30 min, followed by adjustment of the pH to 7-8 and extraction with dichloromethane (100.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (dichloromethane:methanol=40:1 to 10:1) to give the product (520.0 mg, yield: 69.9%).
(R)-4-(4-bromo-2-methoxyphenyl)-N-(1-methylpiperidin-3-yl)phthalazin-1-amine (260.0 mg, 0.60 mmol, 1.0 eq.) was added to dichloromethane (5.0 mL), and the mixture was cooled to −20° C. in nitrogen atmosphere, followed by dropwise addition of boron tribromide (300.6 mg, 1.20 mmol, 2.0 eq.). The resulting mixture was gradually warmed to room temperature and reacted for 2 h. After the reaction was completed as monitored by TLC, methanol (5.0 mL) was added dropwise and slowly under an ice bath to quench the reaction, and the mixture was concentrated under reduced pressure, followed by addition of a saturated aqueous sodium bicarbonate solution (50.0 mL). The resulting mixture was stirred for 30 min, and the pH was adjusted to 7-8, followed by extraction with dichloromethane (50.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by preparative thin-layer chromatography to give the product (90.0 mg, yield 36.2%).
1HNMR (300 MHz, DMSO-d6) δ (ppm): 10.15 (s, 1H), 8.40-8.37 (d, J=9 Hz, 1H), 7.87-7.75 (m, 2H), 7.48-7.45 (d, J=9 Hz, 1H), 7.26-7.23 (d, J=9 Hz, 1H), 7.17-7.07 (m, 3H), 4.41 (s, 1H), 3.09-3.07 (d, J=6 Hz, 1H), 2.73-2.69 (d, J=12 Hz, 1H), 2.22 (s, 3H), 1.99-1.89 (m, 3H), 1.78-1.43 (m, 3H).
Molecular formula: C20H21BrN4O Precise molecular weight: 412.09 LC-MS (m/z)=413.01/415.03[M+H]+.
2-Bromo-5-methoxyphenol (1.8 g, 8.86 mmol, 1.0 eq.) was added into dry tetrahydrofuran (20.0 mL), and the mixture was cooled to 0° C. in nitrogen atmosphere, followed by slow addition of sodium hydride (60%) (531.6 mg, 13.29 mmol, 1.5 eq.). The resulting mixture was reacted for 0.5 h, followed by addition of chloromethoxy ethane (1.17 g, 12.40 mmol, 1.4 eq.). The mixture was warmed to room temperature and reacted for 12 h. After the reaction was completed as monitored by TLC, water (20.0 mL) was added to the system, followed by extraction with ethyl acetate (100.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (specification of silica gel: 100-200 mesh, petroleum ether:ethyl acetate=10:1) to give the product (2.1 g, yield: 91.3%).
1-Bromo-2-(ethoxymethyl)-4-methoxybenzene (2.1 g, 8.04 mmol, 1.0 eq.), bis(pinacolato)diboron (3.06 g, 12.06 mmol, 1.5 eq.), potassium acetate (1.57 g, 16.08 mmol, 2.0 eq.) and Pd(dppf)Cl2 (294.0 mg, 0.40 mmol, 0.05 eq.) were added to 1,4-dioxane (20.0 mL), followed by purge with nitrogen for 5 min. The mixture was reacted at 100° C. for 2 h. After the reaction was completed as monitored by TLC, the reaction solution was purified by silica gel column chromatography (specification of silica gel: 100-200 mesh, petroleum ether: ethyl acetate=10:1) to give the product (2.1 g, yield: 87.5%).
(R)-4-chloro-N-(1-methylpiperidin-3-yl)phthalazin-1-amine (505.0 mg, 1.82 mmol, 1.0 eq.), 2-(2-(ethoxymethoxy)-4-methoxyphenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (843.4 mg, 2.73 mmol, 1.5 eq.), sodium bicarbonate (305.7 mg, 3.64 mmol, 2.0 eq.), water (5.0 mL) and tetrakis(triphenylphosphine)palladium (105.1 mg, 0.09 mmol, 0.05 eq.) were added to 1,4-dioxane (10.0 mL), and the mixture was reacted at 110° C. for 4 h in nitrogen atmosphere. After the reaction was completed as monitored by TLC, water (100.0 mL) was added, followed by extraction with ethyl acetate (100.0 mL). The organic phase was dried and filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (specification of silica gel: 100-200 mesh, dichloromethane:methanol=40:1 to 10:1) to give the product (470.0 mg, yield: 61.1%).
(R)-4-(2-(ethoxymethoxy)-4-methoxyphenyl)-N-(1-methylpiperidin-3-yl)phthalazin-1-amine (470.0 mg, 1.11 mmol, 1.0 eq.) was added to dichloromethane (5.0 mL), followed by dropwise addition of trifluoroacetic acid (3.0 mL). The mixture was reacted at room temperature for 2 h. After the reaction was completed as monitored by TLC, the pH was adjusted to 7-8 with a saturated aqueous sodium bicarbonate solution under an ice bath, followed by extraction with dichloromethane (50.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by preparative thin-layer chromatography (dichloromethane:methanol=10:1) to give the product (50.0 mg, yield: 12.3%).
1HNMR (300 MHz, DMSO-d6) δ (ppm): 9.90 (s, 1H), 8.47 (s, 1H), 7.77-7.88 (m, 2H), 7.55-7.58 (m, 1H), 7.36 (s, 1H), 7.20-7.23 (d, J=6 Hz, 1H), 6.54-6.59 (m, 2H), 4.61 (s, 1H), 3.79 (s, 3H), 3.36 (m, 1H), 3.05 (m, 2H), 2.58 (s, 3H), 1.74-2.04 (m, 5H).
Molecular formula: C21H24N4O2 Molecular weight: 364.45 LC-MS (Pos, m/z): 365.15[M+H]+.
4-Bromo-1-iodo-2-methoxybenzene (21 g, 67.06 mmol, 1.0 eq.) was dissolved in anhydrous THE (200 mL), followed by dropwise addition of a solution of i-PrMgCl LiCl in THE (1.3 mol/L, 55 mL, 70.41 mmol, 1.05 eq.) at −20° C. After the completion of dropwise addition, the mixture was reacted for 30 min. At −20° C., the above reaction solution was added dropwise to a solution of 3,4-pyridine dicarboxylic anhydride (10 g, 67.06 mmol, 1.0 eq.) in THE (100 mL). After the completion of dropwise addition, the mixture was reacted at room temperature for 1 h. After the reaction was completed as detected by LC-MS, the reaction solution was poured into a saturated aqueous ammonium chloride solution (200 mL), followed by concentration. A large number of solids precipitated, and the mixture was filtered. The filter cake was dried to give an isomer-containing mixed product (18.67 g, yield: 82.8%).
The mixture obtained in the previous step (5 g, 14.87 mmol, 1.0 eq.) was added to EtOH (30 mL), followed by addition of 85% hydrazine hydrate (1.76 g, 29.74 mmol, 2.0 eq.). The mixture was heated to 90° C. and reacted for 2 h. After the reaction was completed as detected by TLC, the reaction solution was concentrated, and the crude product was purified by silica gel column chromatography (dichloromethane:methanol=1200:1 to 300:1, v/v) to give the product (3.6 g, yield: 72.8%).
1-(4-Bromo-2-methoxyphenyl)pyrido[3,4-d]pyridazin-4-ol (3 g, 9.03 mmol, 1.0 eq.) was added to acetonitrile (50 mL), followed by addition of phosphorus oxychloride (2.77 g, 18.06 mmol, 2.0 eq.). The mixture was reacted at 100° C. for 16 h. After the reaction was completed as detected by TLC, the reaction solution was concentrated, and the concentrated solution was poured into ice water (10 mL). The pH was adjusted to alkalinity with sodium bicarbonate, followed by extraction with dichloromethane (30 mL×3). The organic phases were combined, dried and concentrated, and the crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=10:1 to 5:1, v/v) to give the product (2.1 g, yield: 66%).
1-(4-Bromo-2-methoxyphenyl)-4-chloropyrido[3,4-d]pyridazine (2.1 g, 5.99 mmol, 1.0 eq.) was dissolved in dichloromethane (20 mL), followed by dropwise and slow addition of a solution of boron tribromide in n-heptane (1 mol/L, 9 mL, 8.96 mmol, 1.5 eq.) at −20° C. After the completion of dropwise addition, the mixture was reacted at room temperature for 10 min. After the reaction was completed as detected by TLC, a saturated sodium bicarbonate solution (10 mL) was added dropwise and slowly to the reaction solution under an ice bath, followed by extraction with dichloromethane (20 mL×3). The organic phases were combined, dried and concentrated, and the crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=2:1, v/v) to give the product (1.52 g, yield: 76%).
5-Bromo-2-(4-chloropyrido[3,4-d]pyridazin-1-yl)phenol (520 mg, 1.55 mmol, 1.0 eq.) was dissolved in DMAc (5 mL), followed by addition of (R)-1-tert-butoxycarbonyl-3-aminopiperidine (621 mg, 3.1 mmol, 2.0 eq.). The mixture was refluxed at 120° C. for 1 h. After the reaction was completed as detected by TLC, the reaction solution was added dropwise into water (5 mL), followed by extraction with ethyl acetate (10 mL×2). The organic phases were combined, dried and concentrated, and the crude product was purified by silica gel column chromatography (dichloromethane:methanol=200:1 to 50:1) to give the product (500 mg, yield: 64.5%).
Tert-butyl (R)-3-((1-(4-bromo-2-methoxyphenyl)pyrido[3,4-d]pyridazin-4-yl)amino)piperidine-1-carboxylate (500 mg, 1 mmol, 1.0 eq.) was dissolved in DMAc (5 mL), followed by addition of zinc cyanide (94 mg, 0.8 mmol, 0.8 eq.) and tetrakis(triphenylphosphine)palladium (112 mg, 0.1 mmol, 0.1 eq.) in nitrogen atmosphere. The mixture was reacted at 120° C. for 45 min. After the reaction was completed as detected by LC-MS, the reaction solution was poured into water, followed by extraction with EA (10 mL×2). The organic phases were combined, dried and concentrated, and the crude product was purified by silica gel column chromatography (dichloromethane:methanol=200:1, v/v) to give the product (350 mg, yield: 78.5%).
Tert-butyl (R)-3-((1-(4-cyano-2-hydroxyphenyl)pyrido[3,4-d]pyridazin-4-yl)amino)piperidine-1-carboxylate (350 mg, 0.784 mmol, 1.0 eq.) was dissolved in EA (1 mL), and the solution was added dropwise into a 4 mol/L solution of hydrogen chloride in 1,4-dioxane (4 mL). After the completion of dropwise addition, a large number of solids precipitated. The mixture was reacted at room temperature for 5 min. After the reaction was completed as detected by TLC, water (2 mL) was added to the reaction solution, the pH was adjusted to alkalinity with sodium bicarbonate, and the aqueous phase was concentrated to give the crude product. The crude product was dissolved in dichloromethane/methanol (20:1, 20 mL), and the solution was filtered. The filtrate was dried and concentrated, and the crude product was directly used in the next step.
(R)-3-hydroxy-4-(4-(piperidin-3-ylamino)pyrido[3,4-d]pyridazin-1-yl)benzonitrile (0.783 mmol, 1.0 eq.) was dissolved in methanol (2 mL), followed by addition of an aqueous formaldehyde solution (37%) (63 mg, 0.783 mmol, 1.0 eq.). The mixture was stirred at room temperature for 5 min, followed by addition of sodium cyanoborohydride (55 mg, 0.862 mmol, 1.1 eq.). The resulting mixture was reacted at room temperature for 5 min. After the reaction was completed as detected by TLC, the reaction solution was concentrated, saturated brine (1 mL) was added, and the pH was adjusted to alkalinity with sodium bicarbonate, followed by extraction with a mixed solvent (dichloromethane:methanol:ammonia solution=20:1:0.5, 20 mL×2). The organic phases were combined, dried and concentrated, and the crude product was purified by silica gel column chromatography (dichloromethane:methanol=50:1 to 10:1, v/v) and lyophilized to give the product (91 mg, two-step yield: 32.2%).
1H-NMR (400 MHz, DMSO-d6) δ (ppm): 10.62 (brs, 1H), 9.84 (s, 1H), 8.87-8.86 (d, 1H), 7.83 (s, 1H), 7.54-7.52 (d, 1H), 7.45-7.43 (m, 1H), 7.38-7.37 (d, 1H), 7.28-7.26 (d, 1H), 4.55 (s, 1H), 2.40 (s, 2H), 2.24 (m, 2H), 2.02 (s, 3H), 1.85 (m, 2H), 1.67 (m, 2H).
Molecular formula: C20H20N6O Precise molecular weight: 360.17 LC-MS (m/z)=361.10 [M+H]+
4-Bromo-3-methoxybenzaldehyde (5.0 g, 23.25 mmol, 1.0 eq.), bis(pinacolato)diboron (8.8 g, 34.87 mmol, 1.5 eq.), potassium acetate (4.5 g, 46.50 mmol, 2.0 eq.) and Pd(dppf)Cl2 (850.6 mg, 1.16 mmol, 0.05 eq.) were added to 1,4-dioxane (30.0 mL), and the mixture was reacted at 100° C. for 4 h in nitrogen atmosphere. After the reaction was completed as monitored by TLC, the crude product was purified by silica gel column chromatography (specification of silica gel: 100-200 mesh, petroleum ether:ethyl acetate=5:1) to give the product (5.5 g, yield: 91.6%).
(R)-4-chloro-N-(1-methylpiperidin-3-yl)phthalazin-1-amine (443.7 mg, 1.60 mmol, 1.0 eq.), 3-methoxy-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzaldehyde (630.3 mg, 2.40 mmol, 1.5 eq.), sodium bicarbonate (268.8 mg, 3.20 mmol, 2.0 eq.) and tetrakis(triphenylphosphine)palladium (92.4 mg, 0.08 mmol, 0.05 eq.) were added to 1,4-dioxane (10.0 mL) and water (5.0 mL), and the mixture was reacted at 110° C. for 4 h in nitrogen atmosphere. After the reaction was completed as monitored by TLC, ethyl acetate (200.0 mL) was added, followed by washing with a saturated aqueous sodium chloride solution (100.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (specification of silica gel: 100-200 mesh, dichloromethane:methanol=40:1 to 10:1) to give the product (410.0 mg, yield: 68.1%).
(R)-3-methoxy-4-(4-((1-methylpiperidin-3-yl)amino)phthalazin-1-yl)benzaldehyde (410.0 mg, 1.08 mmol, 1.0 eq.) was added to dichloromethane (10.0 mL), followed by dropwise addition of boron tribromide (818.5 mg, 3.26 mmol, 3.0 eq.) under an ice bath. The mixture was warmed to room temperature and stirred for 4 h. After the reaction was completed as monitored by TLC, methanol (5.0 mL) was added dropwise under an ice bath to quench the reaction, followed by concentration under reduced pressure, addition of dichloromethane (100.0 mL) and back extraction with water (50.0 mL). The pH of the aqueous phase was adjusted to 7-8 with a saturated aqueous sodium bicarbonate solution, followed by extraction with dichloromethane (100.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by preparative thin-layer chromatography to give the product (260.0 mg, yield: 66.4%).
(R)-3-hydroxy-4-(4-((1-methylpiperidin-3-yl)amino)phthalazin-1-yl)benzaldehyde (260.0 mg, 0.71 mmol, 1.0 eq.) and diethylaminosulfur trifluoride (1.15 g, 7.1 mmol, 10.0 eq.) were added to dichloromethane (10.0 mL), and the mixture was stirred at room temperature for 12 h. After the reaction was completed as monitored by TLC, the reaction solution was added dropwise and slowly to ice water, and the pH was adjusted to 7-8 with a saturated aqueous sodium bicarbonate solution, followed by extraction with dichloromethane (100.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by preparative thin-layer chromatography to give the product (70.0 mg, yield: 25.6%).
1HNMR (300 MHz, DMSO-d6) δ (ppm): 10.10 (s, 1H), 8.41-8.38 (d, J=9 Hz, 1H), 7.87-7.75 (m, 2H), 7.46-7.41 (m, 2H), 7.26 (s, 0.29H), 7.17-7.09 (m, 3H), 7.07 (s, 0.47H), 6.88 (s, 0.25H), 4.44-4.41 (m, 1H), 3.10-3.08 (d, J=6 Hz, 1H), 2.74-2.70 (d, J=12 Hz, 1H), 2.22 (s, 3H), 2.00-1.90 (m, 3H), 1.78-1.44 (m, 3H).
Molecular formula: C21H22F2N4O Precise molecular weight: 384.18 LC-MS (m/z)=385.12[M+H]+.
2-Iodo-5-methylbenzoic acid (20.0 g, 76.32 mmol, 1.0 eq.), iodomethane (16.25 g, 114.48 mmol, 1.5 eq.) and potassium carbonate (15.82 g, 114.48 mmol, 1.5 eq.) were dissolved in DMF (200 mL), and the mixture was reacted at room temperature for 0.5 h. After the reaction was completed as monitored by LC-MS, the reaction solution was poured into water (200 mL), followed by extraction with MTBE (200 mL×2). The organic phase was dried and concentrated to give the product (20 g, yield: 94.9%).
4-Bromo-2-methoxybenzoic acid (40.0 g, 173.12 mmol, 1.0 eq.) and DMF (0.5 mL) were dissolved in DCM (400 mL), followed by dropwise addition of oxalyl chloride (13.18 g, 103.87 mmol, 0.6 eq.). The mixture was reacted for 1 h. After the reaction was completed as monitored by TLC, the reaction solution was concentrated under reduced pressure to give the product (40 g, yield: 92.6%).
Methyl 2-iodo-5-methylbenzoate (20 g, 72.44 mmol, 1.0 eq.) was dissolved in THE (200 mL), and the mixture was cooled to −20° C., followed by addition of a solution of an isopropylmagnesium chloride lithium chloride complex in tetrahydrofuran (1.3 mol/L, 61.3 mL, 79.68 mmol, 1.1 eq.). The resulting mixture was stirred for 1 h, and a solution of 4-bromo-2-methoxybenzoyl chloride (18.07 g, 72.44 mmol, 1.0 eq.) in THE (200 mL) was added dropwise to the system. The mixture was reacted for 20 min. After the reaction was completed as monitored by LC-MS, the reaction solution was poured into water (200 mL), followed by extraction with MTBE (200 mL×2). The organic phase was dried and concentrated to give the product (20 g, yield: 76.0%).
Methyl 2-(4-bromo-2-methoxybenzoyl)-5-methylbenzoate (20.0 g, 55.06 mmol, 1.0 eq.) and 85% hydrazine hydrate (4.13 g, 82.59 mmol, 1.5 eq.) were dissolved in ethanol (100 mL), and the mixture was reacted at 80° C. for 1 h. After the reaction was completed as monitored by LC-MS, the reaction solution was cooled to room temperature and filtered, and the filter cake was dried under reduced pressure to give the product (11 g, yield: 57.8%).
4-(4-Bromo-2-methoxyphenyl)-7-methylphthalazin-1-ol (11.0 g, 31.86 mmol, 1.0 eq.) and phosphorus oxychloride (9.77 g, 63.72 mmol, 2.0 eq.) were dissolved in acetonitrile (110 mL), and the mixture was reacted at 90° C. for 1 h. After the reaction was completed as monitored by LC-MS, the reaction solution was concentrated under reduced pressure, and the crude product was completely dissolved in DCM (300 mL). Water (300 mL) was added, and the pH was adjusted to 9 with NaHCO3, followed by liquid separation. The aqueous phase was extracted with DCM (100 mL), and the organic phases were combined, dried and concentrated to give the product (10 g, yield: 86.3%).
1-(4-Bromo-2-methoxyphenyl)-4-chloro-6-methylphthalazine (10 g, 27.50 mmol, 1.0 eq.), tert-butyl (R)-3-aminopiperidine-1-carboxylate (8.26 g, 41.25 mmol, 1.5 eq.) and DIPEA (10.66 g, 82.50 mmol, 3.0 eq.) were dissolved in DMAC (100 mL), and the mixture was reacted at 120° C. for 12 h. After the reaction was completed as detected by TLC, the reaction solution was poured into water (200 mL), followed by extraction with EA (200 mL×2). The organic phases were combined, washed with water (200 mL×2), dried and concentrated, and the crude product was purified by silica gel column chromatography (PE:EA=2:1) to give the product (11 g, yield: 75.8%).
Tert-butyl (R)-3-((4-(4-bromo-2-methoxyphenyl)-7-methylphthalazin-1-yl)amino)piperidine-1-carboxylate (11 g, 20.85 mmol, 1.0 eq.) was dissolved in DCM (220 mL), followed by dropwise addition of boron tribromide (15.7 g, 62.55 mmol, 3.0 eq.). The mixture was reacted for 1 h. After the reaction was completed as detected by LC-MS, an appropriate amount of methanol was added to the system, and the reaction solution was concentrated under reduced pressure. The crude product was completely dissolved in water (50 mL), followed by liquid separation. The pH of the aqueous phase was adjusted to 9 with NaHCO3, followed by extraction with a mixed solvent of dichloromethane and methanol (10:1, 100 mL×3). The organic phase was dried and concentrated to give the product (6.0 g, yield: 69.6%).
(R)-5-bromo-2-(6-methyl-4-(piperidin-3-ylamino)phthalazin-1-yl)phenol (6.0 g, 14.51 mmol, 1.0 eq.) was dissolved in methanol (60 mL), followed by addition of an aqueous formaldehyde solution (37%) (1.18 g, 14.51 mmol, 1.0 eq.). The mixture was stirred at room temperature for 5 min, followed by addition of sodium cyanoborohydride (911 mg, 14.51 mmol, 1.0 eq.). The mixture was reacted at room temperature for 5 min. After the reaction was completed as monitored by TLC, the reaction solution was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (DCM:MeOH=10:1) to give the product (4.5 g, yield: 72.5%).
1HNMR (300 MHz, DMSO-d6) δ (ppm): 10.20-10.16 (d, 1H), 8.20 (s, 1H), 7.63-7.60 (d, 1H), 7.40-7.37 (d, 1H), 7.25-7.23 (d, 1H), 7.17-7.12 (m, 2H), 7.00-6.97 (d, 1H), 4.41 (s, 1H), 3.11-3.08 (d, 1H), 2.75-2.71 (d, 1H), 2.53 (s, 3H), 2.24 (s, 3H), 2.00-1.96 (d, 3H), 1.80-1.75 (d, 1H), 1.68-1.60 (m, 1H), 1.52-1.45 (m, 1H).
Molecular formula: C21H23BrN4O Molecular weight: 426.11 LC-MS (m/z)=427.01/429.01[M+H]+.
4-Bromo-2-methoxybenzoic acid (25.0 g, 108.20 mmol, 1.0 eq.), iodomethane (23.04 g, 162.30 mmol, 1.5 eq.) and potassium carbonate (22.40 g, 162.30 mmol, 1.5 eq.) were dissolved in DMF (250 mL), and the mixture was reacted at room temperature for 1 h. After the reaction was completed as monitored by TLC, the reaction solution was poured into water (500 mL), followed by extraction with MTBE (500 mL×2). The organic phase was washed with water (500 mL×2), dried and concentrated to give the product (26.5 g, yield: 100%).
Methyl 4-bromo-2-methoxybenzoate (26.5 g, 108.20 mmol, 1.0 eq.), cyclopropylboronic acid (18.59 g, 216.40 mmol, 2.0 eq.), tricyclohexylphosphorus (15.20 g, 54.10 mmol, 0.5 eq.), palladium acetate (2.43 g, 10.82 mmol, 0.1 eq.) and potassium phosphate (45.94 g, 216.40 mmol, 2.0 eq.) were dissolved in toluene (520 mL) and water (100 mL), and the mixture was reacted at 100° C. for 23 h in nitrogen atmosphere. After the reaction was completed as detected by TLC, the reaction solution was concentrated under reduced pressure, the aqueous phase was extracted with EA (200 mL×3), and the organic phase was dried and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (PE:EA=20:1) to give the product (18 g, yield: 80.7%).
Methyl 4-cyclopropyl-2-methoxybenzoate (18 g, 87.27 mmol, 1.0 eq.) and lithium hydroxide monohydrate (7.32 g, 174.54 mmol, 2.0 eq.) were dissolved in MeOH (180 mL) and H2O (90 mL), and the mixture was reacted at 40° C. for 3 h. After the reaction was completed as monitored by TLC, the reaction solution was concentrated under reduced pressure, and the crude product was completely dissolved in water (100 mL). The pH was adjusted to 3 with dilute hydrochloric acid, and the aqueous phase was extracted with DCM (200 mL×2), and the organic phase was washed with a 0.2 mol/L aqueous sodium hydroxide solution (300 mL). The aqueous phases were combined, adjusted to pH of 3 with dilute hydrochloric acid and extracted with DCM (200 mL×2). The organic phase was dried and concentrated to give the product (14 g, yield: 83.4%).
4-Cyclopropyl-2-methoxybenzoic acid (14 g, 72.83 mmol, 1.0 eq.) and DMF (0.15 mL) were dissolved in DCM (280 mL), followed by dropwise addition of oxalyl chloride (5.55 g, 43.70 mmol, 0.6 eq.). The mixture was reacted for 1 h, and the reaction solution was concentrated under reduced pressure to give the product (15.34 g, yield: 100%).
Methyl 5-bromo-2-iodobenzoate (20 g, 58.66 mmol, 1.0 eq.) was dissolved in THE (200 mL), and the mixture was cooled to −30° C., followed by addition of a solution of an isopropylmagnesium chloride lithium chloride complex in tetrahydrofuran (1.3 mol/L, 49.6 mL, 64.53 mmol, 1.1 eq.). The resulting mixture was stirred for 0.5 h, and a solution of 4-cyclopropyl-2-methoxybenzoyl chloride (13.6 g, 64.53 mmol, 1.1 eq.) in THE (100 mL) was added dropwise to the system. The mixture was reacted for 20 min. After the reaction was completed as monitored by TLC, the reaction solution was poured into water (200 mL), followed by extraction with EA (200 mL×2). The organic phase was dried and concentrated to give the product (22.83 g, yield: 100%).
Methyl 5-bromo-2-(4-cyclopropyl-2-methoxybenzoyl)benzoate (22.83 g, 58.66 mmol, 1.0 eq.) and 85% hydrazine hydrate (4.99 g, 117.32 mmol, 2.0 eq.) were dissolved in ethanol (150 mL), and the mixture was reacted at 80° C. for 0.5 h. After the reaction was completed as monitored by TLC, the reaction solution was cooled to room temperature and filtered, and the filter cake was dried to give the product (9 g, yield: 41.5%).
7-Bromo-4-(4-cyclopropyl-2-methoxyphenyl)phthalazin-1-ol (9.0 g, 24.24 mmol, 1.0 eq.) and phosphorus oxychloride (7.43 g, 48.48 mmol, 2.0 eq.) were dissolved in acetonitrile (90 mL), and the mixture was reacted at 90° C. for 3 h. After the reaction was completed as monitored by TLC, the reaction solution was concentrated under reduced pressure, and the crude product was dissolved in EA (300 mL). The pH was adjusted to 9 with an aqueous NaHCO3 solution, followed by liquid separation. The aqueous phase was extracted with EA (50 mL), and the organic phases were combined, dried and concentrated. The crude product was slurried with PE:EA (20:1, 100 mL) and filtered under vacuum, and the filter cake was dried to give the product (8.0 g, yield: 84.7%).
6-Bromo-4-chloro-1-(4-cyclopropyl-2-methoxyphenyl)phthalazine (8.0 g, 20.53 mmol, 1.0 eq.) was dissolved in DCM (160 mL), followed by dropwise addition of a 1 mol/L solution of boron tribromide in dichloromethane (30.8 mL, 30.80 mmol, 1.5 eq.). The mixture was reacted for 1 h. After the reaction was completed as detected by TLC, the reaction solution was poured into saturated NaHCO3 (200 mL), and the mixture was stirred for 10 min, followed by liquid separation. The aqueous phase was extracted with DCM (100 mL), and the organic phases were combined, dried and concentrated to give the product (6.0 g, yield: 77.8%).
2-(6-Bromo-4-chlorophthalazin-1-yl)-5-cyclopropylphenol (6.0 g, 15.97 mmol, 1.0 eq.) and tert-butyl (R)-3-aminopiperidine-1-carboxylate (6.40 g, 91.94 mmol, 2.0 eq.) were dissolved in toluene (500 mL), and the mixture was reacted at 100° C. for 18 h. After the reaction was completed as detected by TLC, the reaction solution was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (PE:EA=2:1) to give the product (2.3 g, yield: 26.7%).
Tert-butyl (R)-3-((7-bromo-4-(4-cyclopropyl-2-hydroxyphenyl)phthalazin-1-yl)amino)piperidine-1-carboxylate (2.0 g, 3.71 mmol, 1.0 eq.) was dissolved in EA (20 mL), followed by dropwise addition of a solution of hydrogen chloride in 1,4-dioxane (4 mol/L, 20 mL). The mixture was reacted at room temperature for 2 h. After the reaction was completed as monitored by LC-MS, the reaction solution was concentrated under reduced pressure, and the crude product was completely dissolved in water (20 mL). The aqueous phase was extracted with EA (20 mL), and the pH of the aqueous phase was adjusted to 9 with NaHCO3, followed by extraction with DCM (20 mL×3). The organic phase was dried and concentrated to give the product (1.3 g, yield: 79.7%).
(R)-2-(6-bromo-4-(piperidin-3-ylamino)phthalazin-1-yl)-5-cyclopropylphenol (1.3 g, 2.95 mmol, 1.0 eq.) was dissolved in methanol (13 mL), followed by addition of an aqueous formaldehyde solution (37%) (240 mg, 2.95 mmol, 1.0 eq.). The mixture was stirred at room temperature for 10 min, followed by addition of sodium cyanoborohydride (186 mg, 2.95 mmol, 1.0 eq.). The mixture was reacted at room temperature for 10 min. After the reaction was completed as monitored by TLC, the reaction solution was concentrated under reduced pressure, and the crude product was dispersed with water (20 mL) and extracted with DCM (20 mL×2). The organic phase was dried and concentrated, and the crude product was purified by silica gel column chromatography (DCM:MeOH=50:1) to give the product (1.0 g, yield: 74.6%).
(R)-2-(6-bromo-4-((1-methylpiperidin-3-yl)amino)phthalazin-1-yl)-5-cyclopropylphenol (500 mg, 1.10 mmol, 1.0 eq.), zinc cyanide (130 mg, 1.10 mmol, 1.0 eq.) and tetrakis(triphenylphosphine)palladium (127 mg, 0.11 mmol, 0.1 eq.) were added to DMAC (10 mL), and the mixture was reacted at 120° C. for 3 h in nitrogen atmosphere. After the reaction was completed as monitored by LC-MS, the reaction solution was poured into water (20 mL), and the aqueous phase was extracted with EA (20 mL×2). The organic phases were combined, washed with water (20 mL×3), dried and concentrated, and the crude product was purified by preparative thin-layer chromatography (MeOH:DCM=1:10) to give the product (150 mg, yield: 34.2%).
1HNMR (400 MHz, DMSO-d6) δ (ppm): 9.6 (s, 1H), 9.08 (s, 1H), 8.13-8.11 (m, 1H), 7.61-7.59 (d, 1H), 7.53 (s, 1H), 7.18-7.16 (d, 1H), 6.71-6.68 (t, 2H), 4.54 (s, 1H), 2.96 (s, 1H), 2.47 (s, 4H), 2.01-1.90 (m, 3H), 1.72-1.59 (t, 3H), 1.23 (s, 1H), 1.01-0.98 (m, 2H), 0.71-0.69 (m, 2H).
Molecular formula: C24H25N5O Precise molecular weight: 399.21 LC-MS (m/z)=400.15[M+H]+.
(R)-2-(6-bromo-4-((1-methylpiperidin-3-yl)amino)phthalazin-1-yl)-5-cyclopropylphenol (500 mg, 1.10 mmol, 1.0 eq.), 50% trimethylboroxine (1.10 g, 4.40 mmol, 4.0 eq.), cesium carbonate (717 mg, 2.20 mmol, 2.0 eq.) and [1,1′-bis(diphenylphosphino)ferrocene]palladium dichloride (80 mg, 0.11 mmol, 0.1 eq.) were dissolved in H2O (2 mL) and 1,4-dioxane (10 mL), and the mixture was reacted at 100° C. for 18 h in nitrogen atmosphere. After the reaction was completed as monitored by LC-MS, the reaction solution was poured into water (20 mL), followed by extraction with EA (20 mL×2). The organic phase was dried and concentrated, and the crude product was purified by preparative thin-layer chromatography (MeOH:DCM=1:10) to give the product (110 mg, yield: 25.7%).
1HNMR (400 MHz, DMSO-d6) δ (ppm): 9.72 (s, 1H), 8.27 (s, 1H), 7.65-7.62 (m, 1H), 7.48-7.46 (d, 1H), 7.27 (s, 1H), 7.17-7.15 (d, 1H), 6.70-6.66 (m, 2H), 4.62 (s, 1H), 3.48 (s, 1H), 3.14 (s, 1H), 2.65 (s, 3H), 2.54 (s, 3H), 1.99-1.91 (m, 3H), 1.83-1.77 (m, 3H), 1.01-0.97 (m, 2H), 0.72-0.69 (m, 2H).
Molecular formula: C24H28N4O1 Precise molecular weight: 388.23 LC-MS (m/z)=389.16[M+H]+.
2-Bromo-5-trifluoromethylphenol (5.00 g, 20.7 mmol, 1.0 eq.) was dissolved in THF (50 mL), and the mixture was cooled to 0° C., followed by portionwise addition of NaH (60%, 2.94 g, 31.1 mmol, 1.5 eq.). The resulting mixture was stirred for 20 min, and chloromethyl ethyl ether (1.24 g, 31.1 mmol, 1.5 eq.) was added. After 4 h of reaction, saturated aqueous NH4Cl solution was added to quench the reaction, followed by extraction with EA (50 mL×2). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to give the product (6.08 g, yield: 98.0%).
1-Bromo-2-(ethoxymethoxy)-4-(trifluoromethyl)benzene (3.00 g, 10.0 mmol, 1.0 eq.) and triisopropyl borate (2.82 g, 15.0 mmol, 1.5 eq.) were dissolved in THF (30 mL), and the mixture was cooled to −60° C. in nitrogen atmosphere, followed by dropwise addition of n-butyllithium (1.6 mol/L THF solution, 9.4 mL, 15.0 mmol, 1.5 eq.). The mixture was reacted for 3 h. a saturated aqueous NH4Cl solution was added to quench the reaction, followed by extraction with EA (50 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was slurried with petroleum ether to give the product (2.20 g, yield: 83.1%).
(2-(Ethoxymethoxy)-4-(trifluoromethyl)phenyl)boronic acid (229 mg, 0.867 mmol, 1.2 eq.), (R)-4-chloro-N-(1-methylpiperidin-3-yl)phthalazin-1-amine (200 mg, 0.723 mmol, 1.0 eq.), Pd(dppf)Cl2 (26.5 mg, 0.0362 mmol, 0.05 eq.) and NaHCO3 (122 mg, 1.45 mmol, 2.0 eq.) were added successively to 1,4-dioxane (4 mL), followed by addition of H2O (2 mL). The mixture was heated to 110° C. and reacted for 3 h in nitrogen atmosphere. The reaction solution was cooled to room temperature, and water (30 mL) was added to quench the reaction, followed by extraction with EA (30 mL×2). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (DCM:MeOH=50:1 to 20:1) to give the product (245 mg, yield: 73.6%).
(R)-4-(2-(ethoxymethoxy)-4-(trifluoromethyl)phenyl)-N-(1-methylpiperidin-3-yl)phthalazin-1-amine (245 mg, 0.532 mmol, 1.0 eq.) was dissolved in DCM (4 mL), followed by dropwise addition of a solution of hydrogen chloride in 1,4-dioxane (4 mol/L, 1.3 mL, 5.32 mmol, 10.0 eq.). The mixture was stirred at room temperature for 1 h. Then water (20 mL) was added to quench the reaction, followed by washing with DCM (20 mL×2). The pH of the aqueous phase was adjusted to 8 with a saturated aqueous NaHCO3 solution, followed by extraction with DCM (20 mL×2). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to give the product (166 mg, yield: 77.5%).
1HNMR (400 MHz, DMSO-d6) δ (ppm): 10.51 (s, 1H), 8.55 (s, 1H), 7.91-7.88 (m, 1H), 7.83-7.80 (m, 1H), 7.59 (s, 1H), 7.51 (d, J=7.8 Hz, 1H), 7.46 (d, J=8.0 Hz, 1H), 7.35 (s, 1H), 7.31 (d, J=7.9 Hz, 1H), 4.73 (s, 1H), 3.56 (s, 2H), 3.22 (s, 2H), 2.73 (s, 3H), 1.99 (s, 2H), 1.83 (d, J=9.2 Hz, 2H).
Molecular formula: C21H21F3N4O Precise molecular weight: 402.17 LC-MS (m/z)=403.11[M+H]+.
3-Bromo-5-methylpyridine (20.0 g, 0.116 mol, 1.0 eq.) and triisopropyl borate (26.1 g, 0.139 mol, 1.2 eq.) were dissolved in THF (200 mL), and the mixture was cooled to −60° C. in nitrogen atmosphere, followed by dropwise addition of n-butyllithium (2.5 mol/L THF solution, 55.6 mL, 0.139 mol, 1.2 eq.). The mixture was reacted for 2 h. Dilute hydrochloric acid was added to quench the reaction, and the pH was adjusted to 3, followed by liquid separation. The aqueous phase was extracted with EA (100 mL×3), the organic phases were combined, dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The resulting crude product was dissolved in MeOH (100 mL), followed by addition of meta-chloroperoxybenzoic acid (16.0 g, 0.0928 mol, 0.8 eq.). After the mixture was stirred at room temperature for 1 h, a saturated aqueous sodium thiosulfate solution was added to quench the reaction, followed by extraction with DCM (100 mL×3). The organic phase was dried over anhydrous sodium sulfate and concentrated, and the crude product was purified by silica gel column chromatography (DCM:MeOH=100:1 to 20:1) to give the product (10.8 g, yield: 85.1%).
5-Methyl-3-hydroxypyridine (10.0 g, 0.0916 mol, 1.0 eq.) and KOH (10.3 g, 0.183 mol, 2.0 eq.) were added to water (500 mL), followed by addition of iodine (23.2 g, 0.0916 mol, 1.0 eq.). The mixture was reacted at room temperature for 16 h. The pH was adjusted to 7 with a saturated aqueous NH4Cl solution, followed by extraction with EA (100 mL×3). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (DCM:MeOH=100:1 to 50:1) to give the product (11.0 g, yield: 51.1%).
2-Iodo-5-methyl-3-hydroxypyridine (5.0 g, 21.3 mmol, 1.0 eq.) was dissolved in DMF (50 mL), followed by addition of iodomethane (4.53 g, 31.9 mmol, 1.5 eq.) and K2CO3 (5.89 g, 42.6 mmol, 2.0 eq.). The mixture was reacted at room temperature for 2 h. Water (250 mL) was added to quench the reaction, followed by extraction with EA (100 mL). The organic phase was washed with water (50 mL×3), dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (DCM:MeOH=100:1 to 50:1) to give the product (4.20 g, yield: 79.3%).
2-Iodo-3-methoxy-5-methylpyridine (4.00 g, 16.1 mmol, 1.0 eq.) was dissolved in THE (40 mL), and the mixture was cooled to −60° C. in nitrogen atmosphere, followed by dropwise addition of n-butyllithium (2.5 mol/L THE solution, 6.44 mL, 16.1 mmol, 1.0 eq.). The resulting mixture was stirred for 30 min. Phthalic anhydride (2.38 g, 16.1 mmol, 1.0 eq.) was dissolved in THE (40 mL), and the mixture was cooled to −60° C. and then added dropwise to the above reaction solution. After the completion of dropwise addition, the resulting mixture was reacted for 1 h. The pH was adjusted to 6 with dilute hydrochloric acid, followed by extraction with EA. The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to give the product (3.20 g, crude product).
2-(3-Methoxy-5-methylpyridinoyl)benzoic acid (3.2 g, 11.8 mmol, 1.0 eq.) was dissolved in DMF (50 mL), followed by addition of K2CO3 (3.26 g, 23.6 mmol, 2.0 eq.) and iodomethane (2.51 g, 17.7 mmol, 1.5 eq.). The mixture was reacted at room temperature for 1 h. Water (200 mL) was added to quench the reaction, followed by extraction with EA (100 mL). The organic phase was washed with water (50 mL×3), dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (DCM:MeOH=100:1 to 50:1) to give the product (1.30 g, two-step yield: 28.4%).
Methyl 2-(3-methoxy-5-methylpyridinoyl)benzoate (1.20 g, 4.21 mmol, 1.0 eq.) was dissolved in EtOH (12 mL), followed by addition of hydrazine hydrate (85%, 496 mg, 8.42 mmol, 2.0 eq.). The mixture was heated to 80° C. and reacted for 1 h. The reaction solution was cooled to room temperature and concentrated, and the crude product was purified by silica gel column chromatography (DCM:MeOH=100:1 to 20:1) to give the product (920 mg, yield: 81.8%).
4-(3-Methoxy-5-methylpyridin-2-yl)phthalazin-1-ol (900 mg, 3.37 mmol, 1.0 eq.) was added to ACN (20 mL), followed by addition of POCl3 (5.17 g, 33.7 mmol, 10.0 eq.). The mixture was heated to 90° C. and reacted for 4 h. The reaction solution was cooled to room temperature, water was added to quench the reaction, and the pH was adjusted to 8 with a saturated aqueous NaHCO3 solution, followed by extraction with EA (50 mL×3). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (DCM:MeOH=100:1 to 50:1) to give the product (610 mg, yield: 63.4%).
1-Chloro-4-(3-methoxy-5-methylpyridin-2-yl)phthalazine (600 mg, 2.10 mmol, 1.0 eq.), tert-butyl (R)-3-aminopiperidine-1-carboxylate (631 mg, 3.15 mmol, 1.5 eq.), Pd2(dba)3 (385 mg, 0.420 mmol, 0.2 eq.), BINAP (523 mg, 0.840 mmol, 0.4 eq.) and Cs2CO3 (1.37 g, 4.20 mmol, 2.0 eq.) were added successively to 1,4-dioxane (20 mL), and the mixture was heated to 90° C. and reacted for 6 h in nitrogen atmosphere. The reaction solution was cooled to room temperature, and water (50 mL) was added to quench the reaction, followed by extraction with EA (30 mL×2). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (DCM:MeOH=100:1 to 20:1) to give the product (485 mg, yield: 51.4%).
Tert-butyl (R)-3-((4-(3-methoxy-5-methylpyridin-2-yl)phthalazin-1-yl)amino)piperidine-1-carboxylate (485 mg, 1.08 mmol, 1.0 eq.) was dissolved in DCM (15 mL), and the mixture was cooled to −40° C., followed by dropwise addition of BBr3 (2.71 g, 10.8 mmol, 10.0 eq.). The mixture was naturally warmed to room temperature and reacted for 20 h. The reaction solution was concentrated, MeOH was added to quench the reaction, and a saturated aqueous NaHCO3 solution was added to adjust the pH to 8, and DCM (30 mL×3) was added for extraction. The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (DCM:MeOH=100:1 to 10:1) to give the product (160 mg, yield: 44.2%).
(R)-5-methyl-2-(4-(piperidin-3-ylamino)phthalazin-1-yl)pyridin-3-ol (160 mg, 0.477 mmol, 1.0 eq.) was dissolved in MeOH (4 mL), followed by addition of an aqueous formaldehyde solution (37%, 116 mg, 1.43 mmol, 3.0 eq.). The mixture was stirred at room temperature for 30 min. NaBH3CN (90.0 mg, 1.43 mmol, 3.0 eq.) was then added and the mixture was reacted for 1 h. A saturated aqueous NaHCO3 solution was added to quench the reaction, followed by extraction with DCM (20 mL×3). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by preparative thin-layer chromatography (DCM:MeOH=10:1) to give the product (70 mg, yield: 42.0%).
1HNMR (400 MHz, DMSO-d6) δ (ppm): 14.29 (s, 1H), 9.63-9.61 (m, 1H), 8.62 (s, 1H), 8.11 (d, J=1.4 Hz, 1H), 7.99-7.94 (m, 2H), 7.82 (s, 1H), 7.25 (d, J=1.1 Hz, 1H), 4.64 (s, 1H), 3.39 (d, J=10.0 Hz, 2H), 3.05 (d, J=9.0 Hz, 1H), 2.57 (s, 4H), 2.34 (s, 3H), 2.01-1.92 (m, 2H), 1.79-1.74 (m, 2H).
Molecular formula: C20H23N5O Precise molecular weight: 349.19 LC-MS (m/z)=350.14[M+H]+.
Methyl 5-fluoro-2-iodobenzoate (5.0 g, 17.85 mmol, 1.0 eq.) was dissolved in THE (50 mL), and the mixture was cooled to −30° C., followed by dropwise and slow addition of a solution of an isopropylmagnesium chloride lithium chloride complex in tetrahydrofuran (1.3 mol/L, 16.5 mL, 21.42 mmol, 1.2 eq.). The resulting mixture was stirred for 30 min. The above reaction solution was added dropwise and slowly to a solution of 2-methoxy-4-methylbenzoyl chloride (3.30 g, 17.85 mmol, 1.0 eq.) in THE (30 mL) at −60° C. After the completion of dropwise addition, the mixture was naturally warmed to room temperature and reacted for 1 h. After the generation of the product as monitored by LC-MS, the reaction solution was poured into water (100 mL), followed by extraction with EA (50 mL×2). The organic phase was dried and concentrated to give the product (5.4 g, yield: 100%).
Methyl 5-fluoro-2-(2-methoxy-4-methylbenzoyl)benzoate (5.4 g, 17.85 mmol, 1.0 eq.) and 85% hydrazine hydrate (1.05 g, 17.85 mmol, 1.0 eq.) were dissolved in ethanol (50 mL), and the mixture was reacted at room temperature for 12 h. After the generation of the product as monitored by LC-MS, the reaction solution was filtered to give the product (1.0 g, yield: 19.7%).
7-Fluoro-4-(2-methoxy-4-methylphenyl)phthalazin-1-ol (1.0 g, 3.51 mmol, 1.0 eq.) and phosphorus oxychloride (1.07 g, 7.02 mmol, 2.0 eq.) were dissolved in acetonitrile (10 mL), and the mixture was reacted at 90° C. for 0.5 h. After the reaction was completed as monitored by LC-MS, the reaction solution was concentrated under reduced pressure, and the crude product was dissolved in EA (20 mL). The pH was adjusted to 9 with a saturated aqueous sodium bicarbonate solution, followed by liquid separation. The aqueous phase was extracted with EA (20 mL), and the organic phases were combined, dried and concentrated to give the product (0.9 g, yield: 84.9%).
4-Chloro-6-fluoro-1-(2-methoxy-4-methylphenyl)phthalazine (0.9 g, 2.97 mmol, 1.0 eq.), tert-butyl (R)-3-aminopiperidine-1-carboxylate (894 mg, 4.46 mmol, 1.5 eq.), Pd2(dba)3 (274 mg, 0.30 mmol, 0.1 eq.), BINAP (367 mg, 0.59 mmol, 0.2 eq.) and cesium carbonate (1.94 g, 5.94 mmol, 2.0 eq.) were added to 1,4-dioxane (20 mL), and the mixture was reacted at 90° C. for 21 h in nitrogen atmosphere. After the reaction was completed as detected by LC-MS, the reaction solution was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (MeOH:DCM=1:80) to give the product (0.8 g, yield: 57.9%).
Tert-butyl (R)-3-((7-fluoro-4-(2-methoxy-4-methylphenyl)phthalazin-1-yl)amino)piperidine-1-carboxylate (0.8 g, 1.71 mmol, 1.0 eq.) was dissolved in DCM (10 mL), followed by dropwise addition of a 1 mol/L solution of boron tribromide in dichloromethane (5.2 mL, 5.13 mmol, 3.0 eq.). The mixture was reacted for 4 h. After the reaction was completed as detected by LC-MS, an appropriate amount of methanol was added to the flask to quench the reaction, and water (5 mL) was added, followed by liquid separation. The pH of the aqueous phase was adjusted to 9 with a saturated aqueous sodium bicarbonate solution, the aqueous phase was extracted with DCM (15 mL×3), and the organic phase was dried and concentrated to give the product (300 mg, yield: 49.8%).
(R)-2-(6-fluoro-4-(piperidin-3-ylamino)phthalazin-1-yl)-5-methylphenol (300 mg, 0.85 mmol, 1.0 eq.) was dissolved in methanol (5 mL), followed by addition of an aqueous formaldehyde solution (37%) (69 mg, 0.85 mmol, 1.0 eq.). The mixture was stirred at room temperature for 10 min, followed by addition of sodium cyanoborohydride (53 mg, 0.85 mmol, 1.0 eq.). The mixture was reacted at room temperature for 10 min. After the reaction was completed as monitored by LC-MS, the reaction solution was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (DCM:MeOH=20:1) to give the product (160 mg, yield: 51.4%).
1HNMR (400 MHz, DMSO-d6) δ (ppm): 9.59 (s, 1H), 8.28-8.25 (m, 1H), 7.69-7.64 (m, 1H), 7.61-7.57 (m, 1H), 7.18-7.16 (d, 1H), 7.03-7.01 (d, 1H), 6.81 (s, 1H), 6.79-6.77 (d, 1H), 4.41-4.39 (d, 1H), 3.14-3.12 (d, 1H), 2.78-2.75 (d, 1H), 2.33 (s, 3H), 2.26 (s, 1H), 2.02-1.99 (d, 3H), 1.79-1.76 (d, 1H), 1.64-1.58 (t, 1H), 1.49-1.41 (m, 1H).
Molecular formula: C21H23FN4O Precise molecular weight: 366.19 LC-MS (Pos, m/z)=367.14[M+H]+.
5-Methylisobenzofuran-1,3-dione (5 g, 30.83 mmol, 1.0 eq.) and 85% hydrazine hydrate (2.73 g, 46.25 mmol, 1.5 eq.) were added to 1,4-dioxane (100 mL) solution, and the mixture was reacted at 80° C. for 1 h. After the reaction was completed as monitored by LC-MS, the reaction solution was filtered under vacuum, and the filter cake was dried to give the product (5.43 g, yield: 100%).
6-Methyl-2,3-dihydro-phthalazine-1,4-dione (5.43 g, 30.83 mmol, 1.0 eq.) and phosphorus oxychloride (18.91 g, 123.32 mmol, 4.0 eq.) were dissolved in acetonitrile (100 mL), and the mixture was reacted at 90° C. for 18 h. The reaction solution was concentrated under reduced pressure, and water (50 mL) and EA (50 mL) were added to the crude product, followed by liquid separation. The aqueous phase was extracted with EA (50 mL×2), and the organic phases were combined, dried and concentrated. The crude product was purified by silica gel column chromatography (PE:EA=10:1) to give the product (2.4 g, yield: 36.4%).
1,4-Dichloro-6-methylphthalazine (1.0 g, 4.69 mmol, 1.0 eq.), 3-methoxy-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzonitrile (1.46 g, 5.63 mmol, 1.2 eq.), PdCl2(dppf) (344 mg, 0.47 mmol, 0.1 eq.) and sodium bicarbonate (788 mg, 9.38 mmol, 2.0 eq.) were dissolved in 1,4-dioxane (20 mL) and H2O (4 mL), and the mixture was reacted at 100° C. for 3 h in nitrogen atmosphere. After the reaction was completed as detected by LC-MS, the reaction solution was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (PE:EA=3:1) to give the product (0.4 g, yield: 29.4%).
The mixture (0.4 g, 1.37 mmol, 1.0 eq.) of 4-(4-hydroxy-6-methylphthalazin-1-yl)-3-methoxybenzonitrile and 4-(4-hydroxy-7-methylphthalazin-1-yl)-3-methoxybenzonitrile and phosphorus oxychloride (0.42 g, 2.74 mmol, 1.0 eq.) were dissolved in acetonitrile (100 mL), and the mixture was reacted at 90° C. for 1.5 h. After the reaction was completed as monitored by TLC, the reaction solution was concentrated under reduced pressure, and the crude product was dissolved in EA (10 mL). A saturated sodium bicarbonate solution was added to adjust the pH to 9, followed by liquid separation. The organic phase was dried and concentrated to give the product (0.4 g, yield: 94.3%).
The mixture (380 mg, 1.23 mmol, 1.0 eq.) of 4-(4-chloro-6-methylphthalazin-1-yl)-3-methoxybenzonitrile and 4-(4-chloro-7-methylphthalazin-1-yl)-3-methoxybenzonitrile, tert-butyl (R)-3-aminopiperidine-1-carboxylate (368 mg, 1.84 mmol, 1.5 eq.), Pd2(dba)3 (110 mg, 0.12 mmol, 0.1 eq.), BINAP (174 mg, 0.25 mmol, 0.2 eq.) and cesium carbonate (802 mg, 2.46 mmol, 2.0 eq.) were added to 1,4-dioxane (10 mL), and the mixture was reacted at 90° C. for 17 h in nitrogen atmosphere. After the reaction was completed as detected by LC-MS, the reaction solution was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (MeOH:DCM=1:50) to give the product (0.3 g, yield: 78.9%).
The mixture (300 mg, 0.63 mmol, 1.0 eq.) of tert-butyl (R)-3-((4-(4-cyano-2-methoxyphenyl)-7-methylphthalazin-1-yl)amino)piperidine-1-carboxylate and tert-butyl (R)-3-((4-(4-cyano-2-methoxyphenyl)-6-methylphthalazin-1-yl)amino)piperidine-1-carboxylate was dissolved in DCM (5 mL), followed by dropwise addition of a 1 mol/L solution of boron tribromide in dichloromethane (1.9 mL, 1.89 mmol, 3.0 eq.) at room temperature. The mixture was reacted at room temperature for 3 h. After the reaction was completed as detected by LC-MS, an appropriate amount of methanol was added to the system to quench the reaction, and water (10 mL) was added, followed by liquid separation. The pH of the aqueous phase was adjusted to 9 with sodium bicarbonate, the aqueous phase was extracted with DCM (15 mL×3), and the organic phase was dried and concentrated to give the product (150 mg, yield: 66.3%).
The mixture (150 mg, 0.42 mmol, 1.0 eq.) of (R)-3-hydroxy-4-(6-methyl-4-(piperidin-3-ylamino)phthalazin-1-yl)benzonitrile and (R)-3-hydroxy-4-(7-methyl-4-(piperidin-3-ylamino)phthalazin-1-yl)benzonitrile and an aqueous formaldehyde solution (37%) (34 mg, 0.42 mmol, 1.0 eq.) were dissolved in methanol (3 mL), and the mixture was stirred at room temperature for 5 min, followed by addition of sodium cyanoborohydride (26 mg, 0.42 mmol, 1.0 eq.). The resulting mixture was reacted at room temperature for 5 min. After the reaction was completed as monitored by LC-MS, the reaction solution was concentrated under reduced pressure, the crude product was dissolved in DCM (10 mL), and water (10 mL) was added, followed by liquid separation. The organic phase was dried and concentrated, and the crude product was purified by preparative thin-layer chromatography (dichloromethane:methanol:ammonia solution=10:1:0.5) to give the product (R)-3-hydroxy-4-(6-methyl-4-((1-methylpiperidin-3-yl)amino)phthalazin-1-yl)benzonitrile with a larger Rf value (23 mg, yield: 14.7%) (compound 55).
1HNMR (400 MHz, DMSO-d6) δ (ppm): 10.48 (s, 1H), 8.21 (s, 1H), 7.62-7.60 (d, 1H), 7.49-7.47 (d, 1H), 7.41-7.39 (t, 1H), 7.34-7.32 (d, 2H), 7.04-7.02 (d, 1H), 4.42-4.40 (t, 1H), 3.08-3.06 (d, 1H), 2.72-2.69 (d, 1H), 2.53 (s, 3H), 2.21 (s, 3H), 1.99-1.90 (m, 3H), 1.78-1.74 (m, 1H), 1.62-1.59 (d, 1H), 1.49-1.43 (m, 1H).
The two-dimensional NOE spectrum showed coupling signals between 7.49-7.47 (d, 1H) and 7.34-7.32 (d, 1H) and coupling signals between 8.21 (s, 1H) and 7.04-7.02 (d, 1H).
Molecular formula: C22H26N4O Precise molecular weight: 373.19 LC-MS (Pos, m/z)=374.14[M+H]+.
Simultaneously, the product (R)-3-hydroxy-4-(7-methyl-4-((1-methylpiperidin-3-yl)amino)phthalazin-1-yl)benzonitrile with a smaller Rf value (20 mg, yield: 12.8%) (compound 54) was also given.
1HNMR (400 MHz, DMSO-d6) δ (ppm): 10.43 (s, 1H), 8.31-8.29 (d, 1H), 7.70-7.68 (d, 1H), 7.48-7.46 (d, 1H), 7.42-7.39 (m, 1H), 7.32 (s, 1H), 7.19 (s, 1H), 7.08-7.06 (d, 1H), 4.41-4.39 (t, 1H), 3.08-3.06 (d, 1H), 2.72-2.69 (d, 1H), 2.42 (s, 3H), 2.21 (s, 3H), 1.98-1.89 (m, 3H), 1.76-1.73 (t, 1H), 1.62-1.59 (d, 1H), 1.49-1.43 (m, 1H).
The two-dimensional NOE spectrum showed coupling signals between 10.43 (s, 1H) and 7.19 (s, 1H), between 8.31-8.29 (d, 1H) and 7.08-7.06 (d, 1H), and between 8.31-8.29 (d, 1H) and 7.70-7.68 (d, 1H).
Molecular formula: C22H26N4O Precise molecular weight: 373.19 LC-MS (Pos, m/z)=374.13[M+H]+.
2-Bromo-5-(trifluoromethyl)benzoic acid (10.0 g, 37.17 mmol, 1.0 eq.), iodomethane (7.91 g, 55.76 mmol, 1.5 eq.) and potassium carbonate (7.69 g, 55.76 mmol, 1.5 eq.) were dissolved in DMF (50 mL), and the mixture was reacted at room temperature for 1 h. After no materials were left as monitored by TLC, the reaction solution was poured into water (50 mL), followed by extraction with MTBE (50 mL×2). The organic phase was dried and concentrated to give the product (10.52 g, yield: 100%).
Methyl 2-bromo-5-(trifluoromethyl)benzoate (8.0 g, 28.26 mmol, 1.0 eq.) was dissolved in THE (50 mL), and the mixture was cooled to −30° C., followed by dropwise and slow addition of a solution of an isopropylmagnesium chloride lithium chloride complex in tetrahydrofuran (1.3 mol/L, 26 mL, 33.91 mmol, 1.2 eq.). The resulting mixture was stirred for 2 h. The system was cooled to −60° C., and a solution of 2-methoxy-4-methylbenzoyl chloride (5.21 g, 28.26 mmol, 1.0 eq.) in THE (50 mL) was added dropwise to the system. The mixture was reacted for 1 h. After the generation of the product as monitored by LC-MS, the reaction solution was poured into a saturated aqueous potassium carbonate solution (200 mL), followed by extraction with EA (100 mL×2). The organic phase was dried and concentrated to give the product (5.0 g, yield: 50.2%).
Methyl 2-(2-methoxy-4-methylbenzoyl)-5-(trifluoromethyl)benzoate (5.0 g, 14.19 mmol, 1.0 eq.) and 85% hydrazine hydrate (1.25 g, 21.28 mmol, 1.5 eq.) were dissolved in ethanol (50 mL), and the mixture was reacted at 80° C. for 17 h. After the reaction was completed as monitored by LC-MS, the reaction solution was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (MEOH:DCM=1:100) to give the product (1.1 g, yield: 23.2%).
4-(2-Methoxy-4-methylphenyl)-7-(trifluoromethyl)phthalazin-1-ol (1.0 g, 2.99 mmol, 1.0 eq.) and phosphorus oxychloride (917 mg, 5.98 mmol, 2.0 eq.) were dissolved in acetonitrile (20 mL), and the mixture was reacted at 90° C. for 1 h. After the reaction was completed as monitored by TLC, the reaction solution was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (PE:EA=10:1) to give the product (0.5 g, yield: 47.6%).
4-Chloro-1-(2-methoxy-4-methylphenyl)-6-(trifluoromethyl)phthalazine (0.5 g, 1.41 mmol, 1.0 eq.), tert-butyl (R)-3-aminopiperidine-1-carboxylate (424 mg, 2.12 mmol, 1.5 eq.), Pd2(dba)3 (128 mg, 0.14 mmol, 0.1 eq.), BINAP (174 mg, 0.28 mmol, 0.2 eq.) and cesium carbonate (919 mg, 2.82 mmol, 2.0 eq.) were added to 1,4-dioxane (10 mL), and the mixture was reacted at 90° C. for 21 h in nitrogen atmosphere. After the reaction was completed as detected by LC-MS, the reaction solution was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (PE:EA=2:1s) to give the product (0.3 g, yield: 41.2%).
Tert-butyl (R)-3-((4-(2-methoxy-4-methylphenyl)-7-(trifluoromethyl)phthalazin-1-yl)amino)piperidine-1-carboxylate (0.3 g, 0.58 mmol, 1.0 eq.) was dissolved in DCM (5 mL), followed by dropwise addition of a 1 mol/L solution of boron tribromide in dichloromethane (1.7 mL, 1.74 mmol, 3.0 eq.). The mixture was reacted for 1 h. After the reaction was completed as detected by LC-MS, an appropriate amount of methanol was added to the system to quench the reaction, and water (5 mL) was added, followed by liquid separation. The pH of the aqueous phase was adjusted to 9 with sodium bicarbonate, followed by extraction with DCM (10 mL×3). The organic phase was dried and concentrated to give the product (100 mg, yield: 42.9%).
(R)-5-methyl-2-(4-(piperidin-3-ylamino)-6-(trifluoromethyl)phthalazin-1-yl)phenol (100 mg, 0.24 mmol, 1.0 eq.) was dissolved in methanol (2 mL), followed by addition of an aqueous formaldehyde solution (37%) (19 mg, 0.24 mmol, 1.0 eq.). The mixture was stirred at room temperature for 5 min, followed by addition of sodium cyanoborohydride (15 mg, 0.24 mmol, 1.0 eq.). The mixture was reacted at room temperature for 5 min. After the reaction was completed as monitored by LC-MS, the reaction solution was concentrated under reduced pressure. The crude product was purified by preparative thin-layer chromatography (DCM:MeOH=8:1) to give the product (20 mg, yield: 20%).
1HNMR (300 MHz, DMSO-d6) δ (ppm): 9.56 (s, 1H), 8.90 (s, 1H), 8.09-8.06 (d, 1H), 7.68-7.66 (d, 1H), 7.50-7.47 (d, 1H), 7.20-7.18 (d, 1H), 6.82-6.78 (d, 1H), 4.46-4.44 (d, 1H), 3.17-3.13 (d, 1H), 2.34 (s, 3H), 2.26 (s, 1H), 2.02-1.98 (d, 3H), 1.82-1.77 (d, 1H), 1.65-1.62 (d, 1H), 1.51-1.45 (m, 1H).
Molecular formula: C22H23F3N4O Precise molecular weight: 416.18 LC-MS (Pos, m/z)=417.13[M+H]+.
Tert-butyl (R)-3-hydroxypiperidine-1-carboxylate (287.3 mg, 1.42 mmol, 1.5 eq.) was added to anhydrous N,N-dimethylformamide (3.0 mL), followed by addition of sodium hydride (60%) (56.8 mg, 1.42 mmol, 1.5 eq.) under an ice bath. The mixture was reacted for 0.5 h in nitrogen atmosphere, followed by dropwise addition of a solution of 2-(4-bromophthalazin-1-yl)-5-methylphenol (300.0 mg, 0.95 mmol, 1.0 eq.) in N,N-dimethylformamide (5.0 mL). The resulting mixture was reacted at room temperature for 12 h. After the reaction was completed as monitored by TLC, water (50.0 mL) was added to the system to quench the reaction, followed by extraction with ethyl acetate (100.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to give the crude product (360.0 mg, used directly in the next step).
Tert-butyl (R)-3-((4-(2-hydroxy-4-methylphenyl)phthalazin-1-yl)oxy)piperidine-1-carboxylate (360.0 mg, crude product, 0.82 mmol, 1.0 eq.) was added to dichloromethane (5.0 mL), followed by dropwise addition of trifluoroacetic acid (5.0 mL). The mixture was reacted at room temperature for 2 h. After the reaction was completed as monitored by TLC, a saturated aqueous sodium bicarbonate solution was added to the system to adjust the pH to 7-8, followed by extraction with dichloromethane (50.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by preparative thin-layer chromatography (specification of silica gel: 100-200 mesh, dichloromethane:methanol=10:1) to give the product (51.0 mg, yield: 18.5%).
(R)-5-methyl-2-(4-(piperidin-3-yloxy)phthalazin-1-yl)phenol (51.0 mg, 0.15 mmol, 1.0 eq.) and an aqueous formaldehyde solution (37%) (14.8 mg, 0.18 mmol, 1.2 eq.) were added to methanol (5.0 mL), and the mixture was stirred at room temperature for 0.5 h, followed by addition of sodium cyanoborohydride (13.2 mg, 0.21 mmol, 1.4 eq.). The resulting mixture was reacted at room temperature for 2 h. After the reaction was completed as monitored by TLC, the reaction solution was concentrated under reduced pressure, followed by addition of a saturated aqueous sodium bicarbonate solution (100.0 mL). The mixture was stirred for 0.5 h and extracted with dichloromethane (100.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by preparative thin-layer chromatography (dichloromethane:methanol=10:1) to give the product (30.0 mg, yield: 57.2%).
1HNMR (300 MHz, DMSO-d6) δ (ppm): 9.58 (s, 1H), 8.22-8.20 (d, J=6 Hz, 1H), 7.98-7.87 (m, 2H), 7.61-7.58 (m, 1H), 7.20-7.18 (m, 1H), 6.84-6.79 (m, 2H), 5.49 (s, 1H), 3.36-3.34 (m, 1H), 3.04-3.01 (m, 1H), 2.55-2.49 (m, 2H), 2.35 (s, 3H), 2.26 (s, 3H), 1.87-1.66 (m, 4H).
Molecular formula: C21H23N3O2 Precise molecular weight: 349.18 LC-MS (m/z)=350.17[M+H]+.
(R)-4-chloro-N-(1-methylpiperidin-3-yl)phthalazin-1-amine (350.0 mg, 1.26 mmol, 1.0 eq.), (4-cyclopropyl-2-methoxyphenyl)boronic acid (364.2 mg, 1.89 mmol, 1.5 eq.), sodium bicarbonate (317.5 mg, 3.78 mmol, 3.0 eq.) and Pd(PPh3)4 (72.8 mg, 0.06 mmol, 0.05 eq.) were added to a mixed solution of 1,4-dioxane (12.0 mL) and water (6.0 mL), and the mixture was reacted at 110° C. for 12 h in nitrogen atmosphere. After the reaction was completed as monitored by TLC, the reaction solution was washed with water (50.0 mL), followed by extraction with ethyl acetate (100.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (specification of silica gel: 100-200 mesh, dichloromethane:methanol=40:1 to 10:1) to give the product (387.0 mg, yield: 79.1%).
(R)-4-(4-cyclopropyl-2-methoxyphenyl)-N-(1-methylpiperidin-3-yl)phthalazin-1-amine (387.0 mg, 0.99 mmol, 1.0 eq.) was added to dichloromethane (5.0 mL), followed by dropwise addition of boron tribromide (748.6 mg, 2.98 mmol, 3.0 eq.) under an ice bath. The reaction mixture was warmed to room temperature and reacted for 2 h. After the reaction was completed as monitored by TLC, methanol (10.0 mL) was added to the system under an ice bath to carefully quench the reaction, followed by concentration under reduced pressure, addition of dichloromethane (100.0 mL) and back extraction with water (50.0 mL). The pH of the aqueous phase was adjusted to 7-8 with a saturated aqueous sodium bicarbonate solution, followed by extraction with dichloromethane (100.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by preparative thin-layer chromatography to give the product (160.0 mg, yield: 43.2%).
1HNMR (300 MHz, DMSO-d6) δ (ppm): 9.66 (s, 1H), 8.47-8.45 (d, J=6 Hz, 1H), 7.88-7.76 (m, 2H), 7.55-7.53 (m, 1H), 7.32 (s, 1H), 7.17-7.15 (m, 1H), 6.70-6.66 (m, 2H), 4.59 (s, 1H), 3.06-3.03 (m, 2H), 2.55 (s, 3H), 2.01-1.70 (m, 6H), 1.02-0.96 (m, 2H), 0.88-0.84 (m, 1H), 0.72-0.67 (m, 2H).
Molecular formula: C23H26N4O Precise molecular weight: 374.21 LC-MS (m/z)=375.16[M+H]+.
4-Cyclopropyl-2-methoxybenzoic acid (5 g, 26.01 mmol, 1.0 eq.) was dissolved in dichloromethane (50 mL), and N,N-dimethylformamide (0.05 mL) was added dropwise, followed by dropwise and slow addition of oxalyl chloride (9.9 g, 78.03 mmol, 3.0 eq.). After the completion of dropwise addition, the mixture was reacted at room temperature for 20 min. After the reaction was completed as detected by TLC, the reaction solution was concentrated and directly used in the next step.
4-Cyclopropyl-2-methoxybenzoyl chloride (7.74 g, 26.01 mmol, 1.0 eq.) was dissolved in tetrahydrofuran (80 mL), followed by purge with nitrogen. A 1.3 mol/L solution of an isopropylmagnesium chloride lithium chloride complex in tetrahydrofuran (22 mL, 28.61 mmol, 1.1 eq.) was added dropwise and slowly at −40° C. After the completion of dropwise addition, the mixture was stirred at −40° C. for 20 min. Cuprous cyanide (2.33 g, 26.01 mmol, 1.0 eq.) was then added, and the mixture was reacted at −40° C. for 30 min. A solution of 4-cyclopropyl-2-methoxybenzoyl chloride (26.01 mmol, 1.0 eq.) in tetrahydrofuran (50 mL) was then added dropwise. After the completion of dropwise addition, the mixture was reacted at 5° C. for 1 h. After the reaction was completed as detected by TLC, saturated brine (100 mL) and a saturated aqueous sodium bicarbonate solution (50 mL) were added to the reaction solution, followed by extraction with ethyl acetate (50 mL×3). The organic phases were combined, dried over anhydrous sodium sulfate and concentrated to give the product (7.37 g, two-step yield: 82%).
Methyl 2-chloro-4-(4-cyclopropyl-2-methoxybenzoyl)nicotinate (7.37 g, 21.32 mmol, 1.0 eq.) was dissolved in methanol (100 mL), followed by addition of triethylamine (6.47 g, 63.96 mmol, 2.0 eq.) and 5% Pd/C (3.6 g) and purge with hydrogen. The mixture was reacted at room temperature of 25° C. for 1 h. After the reaction was completed as detected by TLC, the reaction solution was filtered through celite, and the filtrate was concentrated and purified by silica gel column chromatography (ethyl acetate:petroleum ether=1:5) to give the product (4.06 g, yield: 61.1%).
Methyl 4-(4-cyclopropyl-2-methoxybenzoyl)nicotinate (4.06 g, 13.01 mmol, 1.0 eq.) was added to ethanol (50 mL), followed by addition of 85% hydrazine hydrate (1.53 g, 26.02 mmol, 2.0 eq.). The mixture was heated to 90° C. and reacted for 2 h. After the reaction was completed as detected by TLC, the reaction solution was filtered, and the filter cake was rinsed with ethanol (5 mL) and dried to give the product (2.44 g, yield: 64%). (2100054-002).
1-(4-Cyclopropyl-2-methoxyphenyl)pyrido[3,4-d]pyridazin-4(3H)-one (2.44 g, 8.32 mmol, 1.0 eq.) was added to acetonitrile (30 mL), followed by addition of phosphorus oxychloride (2.55 g, 16.64 mmol, 2.0 eq.). The mixture was reacted at 100° C. for 16 h. After the reaction was completed as detected by TLC, the reaction solution was concentrated and dissolved in dichloromethane (20 mL). The solution was added dropwise to a saturated aqueous sodium bicarbonate solution (100 mL), followed by extraction. The aqueous phase was extracted with dichloromethane (50 mL×3), and the organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous magnesium sulfate and concentrated. The crude product was purified by silica gel column chromatography (ethyl acetate:petroleum ether=1:10 to 1:5) to give the product (1.94 g, yield: 74.9%).
4-Chloro-1-(4-cyclopropyl-2-methoxyphenyl)pyrido[3,4-d]pyridazine (1.94 g, 6.22 mmol, 1.0 eq.) was dissolved in dichloromethane (20 mL), followed by dropwise and slow addition of boron tribromide (4.67 g, 18.66 mmol, 3.0 eq.) at 0° C. After the completion of dropwise addition, the mixture was reacted at room temperature of 25° C. for 30 min. After the reaction was completed as detected by TLC, the reaction solution was added dropwise and slowly to a saturated aqueous sodium bicarbonate solution (50 mL), followed by extraction with dichloromethane (20 mL×3). The organic phases were combined, dried over anhydrous magnesium sulfate and concentrated to give the product (1.2 g).
2-(4-Chloropyrido[3,4-d]pyridazin-1-yl)-5-cyclopropyl-phenol (1.2 g, 4.03 mmol, 1.0 eq.) was dissolved in N,N-dimethylacetamide (12 mL), followed by addition of (R)-1-tert-butoxycarbonyl-3-aminopiperidine (1.61 g, 8.06 mmol, 2.0 eq.). The mixture was reacted at 120° C. for 16 h. After the reaction was completed as detected by TLC, the reaction solution was poured into water (20 mL), followed by extraction with ethyl acetate (20 mL). The organic phase was dried over anhydrous sodium sulfate and concentrated. The crude product was subjected to silica gel column chromatography (ethyl acetate:petroleum ether=1:3 to 1:1) to give the product (1.19 g, two-step yield: 41.4%).
Tert-butyl (R)-3-((1-(4-cyclopropyl-2-methoxyphenyl)pyrido[3,4-d]pyridazin-4-yl)amino)piperidine-1-carboxylate (1.19 g, 2.58 mmol, 1.0 eq.) was dissolved in dichloromethane (3 mL), and the resulting solution was added dropwise to a 4 moL/L solution of hydrogen chloride in 1,4-dioxane (15 mL). After the completion of dropwise addition, the mixture was reacted at room temperature for 5 min. After the reaction was completed as detected by TLC, water (15 mL) was added to the reaction solution, and the pH was adjusted to low alkalinity with sodium bicarbonate, followed by extraction with a mixed solvent of methanol and dichloromethane (1:5, 30 mL×3). The organic phases were combined, dried over anhydrous magnesium sulfate and concentrated, and the crude product was directly used in the next step.
(R)-5-cyclopropyl-2-(4-(piperidin-3-ylamino)pyrido[3,4-d]pyridazin-1-yl)phenol (2.58 mmol, 1.0 eq.) was dissolved in methanol (10 mL), followed by addition of an aqueous formaldehyde solution (37%) (210 mg, 2.58 mmol, 1.0 eq.). The mixture was stirred at room temperature for 5 min, followed by addition of sodium cyanoborohydride (179 mg, 2.84 mmol, 1.1 eq.). The resulting mixture was reacted at room temperature for 5 min. After the reaction was completed as detected by TLC, the reaction solution was concentrated, and saturated brine (2 mL) was added, followed by extraction with a 7 mol/L solution of ammonia in methanol/dichloromethane (1:7, 10 mL×3). The organic phases were combined, dried and concentrated, and the crude product was purified by preparative thin-layer chromatography to give the product (265 mg, two-step yield: 27%).
1H-NMR (400 MHz, DMSO-d6) δ (ppm): 9.79 (s, 1H), 9.65 (s, 1H), 8.85-8.84 (d, 1H), 7.63-7.61 (d, 1H), 7.34-7.32 (d, 1H), 7.20-7.18 (d, 1H), 6.71-6.68 (m, 2H), 4.51 (s, 1H), 3.22 (s, 1H), 2.85-2.83 (d, 1H), 2.30 (s, 3H), 2.12-2.01 (m, 2H), 1.97-1.90 (m, 1H), 1.81-1.52 (m, 4H), 1.02-0.97 (m, 2H), 0.72-0.68 (m, 2H).
Molecular formula: C22H25N5O Precise molecular weight: 375.21 LC-MS (m/z): 376.12 [M+H]+
(R)-4-chloro-N-(piperidin-3-yl)phthalazin-1-amine (505.0 mg, 1.92 mmol, 1.0 eq.), (1-methoxycyclopropoxy)trimethylsilane (1.34 g, 7.68 mmol, 4.0 eq.), acetic acid (1.0 mL), cesium fluoride (291.9 mg, 1.92 mmol, 1.5 eq.) and sodium cyanoborohydride (603.8 mg, 9.61 mmol, 5.0 eq.) were added to methanol (15.0 mL), and the mixture was reacted at 60° C. for 12 h. After the reaction was completed as monitored by TLC, a saturated aqueous sodium bicarbonate solution (50.0 mL) was added to the system, followed by extraction with dichloromethane (100.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (dichloromethane:methanol=80:1 to 50:1) to give the product (350.0 mg, yield: 60.1%).
(R)-4-chloro-N-(1-cyclopropylpiperidin-3-yl)phthalazin-1-amine (350.0 mg, 1.15 mmol, 1.0 eq.), 3-(ethoxymethoxy)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzonitrile (700.8 mg, 2.31 mmol, 2.0 eq.), sodium bicarbonate (291.0 mg, 3.46 mmol, 3.0 eq.) and Pd(dppf)Cl2 (42.2 mg, 0.05 mmol, 0.05 eq.) were added to 1,4-dioxane (10.0 mL) and water (5.0 mL), and the mixture was reacted at 110° C. for 5 h in nitrogen atmosphere. After the reaction was completed as monitored by TLC, the system was cooled to room temperature, and water (50.0 mL) was added, followed by extraction with ethyl acetate (100.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (100-200 mesh silica gel, dichloromethane:methanol=80:1 to 60:1) to give the product (300.0 mg, yield: 58.5%).
(R)-4-(4-((1-cyclopropylpiperidin-3-yl)amino)phthalazin-1-yl)-3-(ethoxymethoxy)benzonitrile (290.0 mg, 0.65 mmol, 1.0 eq.) was added to ethanol (5.0 mL) and a hydrogen chloride-ethanol solution (2.0 mol/L, 10.0 mL), and the mixture was reacted at room temperature for 1 h. After the reaction was completed as monitored by TLC, the pH of the system was adjusted to 7-8 with a saturated aqueous sodium bicarbonate solution, followed by extraction with dichloromethane (50.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (100-200 mesh silica gel, dichloromethane:methanol=80:1 to 40:1) to give the product (190.0 mg, yield: 75.3%).
1HNMR (400 MHz, DMSO-d6) δ (ppm): 10.47 (s, 1H), 8.40-8.38 (d, J=8 Hz, 1H), 7.86 (t, J=8 Hz, 1H), 7.78 (t, J=8 Hz, 1H), 7.51-7.49 (d, J=8 Hz, 1H), 7.43 (s, 1H), 7.41-7.33 (m, 2H), 7.16-7.14 (d, J=8 Hz, 1H), 4.36-4.34 (m, 1H), 3.29-3.27 (m, 1H), 2.92-2.89 (m, 1H), 2.22-2.17 (m, 2H), 2.01-1.99 (m, 1H), 1.74-1.66 (m, 2H), 1.57-1.44 (m, 2H), 0.46-0.34 (m, 4H).
Molecular formula: C23H23N5O Precise molecular weight: 385.19 LC-MS (Pos, m/z)=386.11[M+H]+.
1-Bromo-4-iodo-2-methoxybenzene (6.0 g, 19.17 mmol, 1.0 eq.) was added to dichloromethane (20.0 mL), followed by dropwise and slow addition of boron tribromide (14.4 g, 57.50 mmol, 3.0 eq.). The mixture was reacted at room temperature for 5 h. After the reaction was completed as monitored by TLC, methanol was added to quench the reaction under an ice bath, and the reaction solution was concentrated under reduced pressure, followed by addition of water (50 mL) and extraction with dichloromethane (200.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to give the product (5.3 g, yield: 92.4%).
2-Bromo-5-iodophenol (5.3 g, 17.73 mmol, 1.0 eq.) was added into anhydrous tetrahydrofuran (20.0 mL), followed by addition of sodium hydride (60% by mass, 1.0 g, 26.59 mmol, 1.5 eq.) under an ice bath. The mixture was reacted for 0.5 h, and then chloromethyl ethyl ether (2.5 g, 26.59 mmol, 1.5 eq.) was added. The mixture was warmed to room temperature and reacted for 12 h. After the reaction was completed as monitored by TLC, a saturated aqueous ammonium chloride solution (100.0 mL) was added to the system, followed by extraction with ethyl acetate (200.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (100-200 mesh silica gel, petroleum ether:ethyl acetate=100:1) to give the product (6.15 g, yield: 97.3%).
1-Bromo-2-(ethylmethoxy)-4-iodobenzene (6.15 g, 17.22 mmol, 1.0 eq.) was added to anhydrous tetrahydrofuran (20.0 mL), and the mixture was cooled to −30° C., followed by dropwise addition of a solution of an isopropylmagnesium chloride lithium chloride complex in tetrahydrofuran (1.3 mol/L, 15.9 mL, 20.67 mmol, 1.2 eq.). The resulting mixture was reacted for 0.5 h, and then 1,2-dimethyldisulfane (1.78 g, 18.95 mmol, 1.1 eq.) was added. The mixture was warmed to room temperature and stirred for 2 h. After the reaction was completed as monitored by TLC, a saturated aqueous ammonium chloride solution (100.0 mL) was added to the system, followed by extraction with ethyl acetate (200.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to give the product (4.5 g, yield: 94.3%).
(4-Bromo-3-(ethoxymethoxy)phenyl)(methyl)sulfane (4.5 g, 16.23 mmol, 1.0 eq.), 4,4,5,5-tetramethyl-1,3,2-dioxaborolane (6.18 g, 24.35 mmol, 1.5 eq.), potassium acetate (3.18 g, 32.46 mmol, 2.0 eq.) and Pd(dppf)Cl2 (593.9 mg, 0.81 mmol, 0.05 eq.) were added to 1,4-dioxane (25.0 mL), and the mixture was stirred at 110° C. for 6 h in nitrogen atmosphere. After the reaction was completed as monitored by TLC, water (200.0 mL) was added, followed by extraction with ethyl acetate (200.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (100-200 mesh silica gel, petroleum ether:ethyl acetate=10:1) to give the product (4.8 g, yield: 91.2%).
(R)-4-chloro-N-(1-methylpiperidin-3-yl)phthalazin-1-amine (1.0 g, 3.61 mmol, 1.0 eq.), 2-(2-(ethoxymethoxy)-4-(methylthio)phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (1.75 g, 5.41 mmol, 1.5 eq.), sodium bicarbonate (607.0 mg, 7.22 mmol, 2.0 eq.) and Pd(dppf)Cl2 (132.1 mg, 0.18 mmol, 0.05 eq.) were added to a mixed solution of 1,4-dioxane (15.0 mL) and water (7.0 mL), and the mixture was stirred at 110° C. for 6 h in nitrogen atmosphere. After the reaction was completed as monitored by TLC, water (100.0 mL) was added, followed by extraction with ethyl acetate (100.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (100-200 mesh silica gel, dichloromethane:methanol=40:1 to 10:1) to give the product (1.2 g, yield: 75.9%).
(R)-4-(2-(ethoxymethoxy)-4-(methylthio)phenyl)-N-(1-methylpiperidin-3-yl)phthalazin-1-amine (100.0 mg, 0.22 mmol, 1.0 eq.) and a hydrogen chloride-1,4-dioxane solution (4.0 mol/L, 3.0 mL) were added to dichloromethane (1.0 mL), and the mixture was reacted at room temperature for 2 h. After the reaction was completed as monitored by TLC, the pH of the system was adjusted to 7-8 with a saturated aqueous sodium bicarbonate solution, followed by extraction with dichloromethane (50.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by preparative thin-layer chromatography to give the product (60.0 mg, yield: 69.7%).
1HNMR (400 MHz, DMSO-d6) δ (ppm): 9.92 (s, 1H), 8.38-8.36 (d, J=8 Hz, 1H), 7.84 (t, J=8 Hz, 1H), 7.77 (t, J=8 Hz, 1H), 7.54-7.52 (d, J=8 Hz, 1H), 7.25-7.23 (d, J=8 Hz, 1H), 7.04-7.02 (d, J=8 Hz, 1H), 6.86-6.83 (m, 2H), 4.40 (s, 1H), 3.08-3.06 (d, J=8 Hz, 1H), 2.72-2.69 (d, J=12 Hz, 1H), 2.51 (s, 3H), 2.21 (s, 3H), 1.99-1.88 (m, 3H), 1.77-1.41 (m, 3H).
Molecular formula: C21H24N4OS Precise molecular weight: 380.17 LC-MS (Pos, m/z)=381.03[M+H]+.
1,4-Dichlorophthalazine (157.0 mg, 0.78 mmol, 1.0 eq.), (R)-tetrahydro-2H-pyran-3-amine hydrochloride (162.8 mg, 1.18 mmol, 1.5 eq.) and diisopropylethylamine (203.6 mg, 1.57 mmol, 2.0 eq.) were added to N,N-dimethylacetamide (3.0 mL), and the mixture was stirred at 120° C. for 12 h. After the reaction was completed as monitored by TLC, the system was cooled to room temperature, and water (100.0 mL) was added, followed by extraction with EA (100.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by preparative thin-layer chromatography to give the product (67.0 mg, yield: 32.2%).
(R)-4-chloro-N-(tetrahydro-2H-pyran-3-yl)phthalazin-1-amine (67.0 mg, 0.25 mmol, 1.0 eq.), 3-(ethoxymethoxy)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzonitrile (154.0 mg, 0.50 mmol, 2.0 eq.), sodium bicarbonate (64.0 mg, 0.76 mmol, 3.0 eq.) and Pd(dppf)Cl2 (9.29 mg, 0.01 mmol, 0.05 eq.) were added to 1,4-dioxane (4.0 mL) and water (2.0 mL), and the mixture was reacted at 110° C. for 5 h in nitrogen atmosphere. After the reaction was completed as monitored by TLC, the system was cooled to room temperature, and water (50.0 mL) was added, followed by extraction with ethyl acetate (100.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by preparative thin-layer chromatography to give the product (60.0 mg, yield: 58.4%).
(R)-3-(ethoxymethoxy)-4-(4-((tetrahydro-2H-pyran-3-yl)amino)phthalazin-1-yl)benzonitrile (60.0 mg, 0.14 mmol, 1.0 eq.) and a hydrogen chloride-ethanol solution (2.0 mol/L, 4.0 mL) were added to ethanol (2.0 mL), and the mixture was reacted at room temperature for 2 h. After the reaction was completed as monitored by TLC, the pH of the system was adjusted to 7-8 with saturated sodium bicarbonate, followed by extraction with dichloromethane (50.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to give the product (30.0 mg, yield: 58.5%).
1HNMR (400 MHz, DMSO-d6) δ (ppm): 10.47 (s, 1H), 8.43-8.40 (d, J=12 Hz, 1H), 7.87 (t, J=8 Hz, 1H), 7.79 (t, J=8 Hz, 1H), 7.51-7.49 (d, J=8 Hz, 1H), 7.43 (s, 1H), 7.41 (s, 1H), 7.33 (s, 1H), 7.23-7.21 (d, J=8 Hz, 1H), 4.44-4.39 (m, 1H), 4.09-4.06 (m, 1H), 3.85-3.82 (d, J=12 Hz, 1H), 3.28-3.16 (m, 2H), 2.11 (s, 1H), 1.79-1.65 (m, 3H).
Molecular formula: C20H18N4O2 Precise molecular weight: 346.14 LC-MS (Pos, m/z)=347.02[M+H]+.
4-(4-Chlorophthalazin-1-yl)-3-(ethoxymethoxy)benzonitrile (292.0 mg, 0.85 mmol, 1.0 eq.), tert-butyl (3R,5R)-3-amino-5-fluoropiperidine-1-carboxylate (206.3 mg, 0.94 mmol, 1.1 eq.), cesium carbonate (559.7 mg, 1.71 mmol, 2.0 eq.), Pd2(dba)3 (78.6 mg, 0.08 mmol, 0.1 eq.) and 1,1′-binaphthyl-2,2′-bisdiphenylphosphine (106.9 mg, 0.17 mmol, 0.2 eq.) were added to 1,4-dioxane (15.0 mL), and the mixture was reacted at 90° C. for 12 h in nitrogen atmosphere. After the reaction was completed as monitored by TLC, water (100.0 mL) was added, followed by extraction with ethyl acetate (100.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (100-200 mesh silica gel, dichloromethane:methanol=100:1 to 60:1) to give the product (220.0 mg, yield: 49.1%).
Tert-butyl (3R,5R)-3-((4-(4-cyano-2-(ethoxymethoxy)phenyl)phthalazin-1-yl)amino)-5-fluoropiperidine-1-carboxylate (220.0 mg, 0.42 mmol, 1.0 eq.) and a hydrogen chloride-1,4-dioxane solution (4.0 mol/L, 5.0 mL) were added to dichloromethane (3.0 mL), and the mixture was stirred at room temperature for 1 h. After the reaction was completed as monitored by TLC, the pH of the system was adjusted to 7-8 with a saturated aqueous sodium bicarbonate solution, followed by extraction with dichloromethane (100.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to give the product (150.0 mg, yield: 98.0%).
4-(4-(((3R,5R)-5-fluoropiperidin-3-yl)amino)phthalazin-1-yl)-3-hydroxybenzonitrile (150.0 mg, 0.41 mmol, 1.0 eq.) and an aqueous formaldehyde solution (37% by mass, 50.2 mg, 0.61 mmol, 1.5 eq.) were added to methanol (6.0 mL), and the mixture was stirred at room temperature for 0.5 h, followed by addition of sodium cyanoborohydride (38.8 mg, 0.61 mmol, 1.5 eq.). The resulting mixture was reacted at room temperature for 2 h. After the reaction was completed as monitored by TLC, a saturated aqueous sodium bicarbonate solution (50.0 mL) was added, followed by extraction with dichloromethane (100.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by preparative thin-layer chromatography to give the product (80.0 mg, yield: 51.6%).
1HNMR (400 MHz, DMSO-d6) δ (ppm): 10.47 (s, 1H), 8.40-8.38 (d, J=8 Hz, 1H), 7.90-7.86 (m, 1H), 7.81-7.78 (m, 1H), 7.51-7.49 (d, J=8 Hz, 1H), 7.44-7.41 (m, 2H), 7.33 (s, 1H), 7.25-7.23 (d, J=8 Hz, 1H), 5.07-4.96 (m, 1H), 4.75-4.74 (m, 1H), 3.11-3.09 (d, J=8 Hz, 1H), 2.97-2.92 (m, 1H), 2.24 (s, 3H), 2.21 (s, 1H), 2.14-2.11 (m, 1H), 2.02-1.73 (m, 2H).
Molecular formula: C21H20FN5O Precise molecular weight: 377.17 LC-MS (Pos, m/z)=378.11[M+H]+.
4-Chloro-2-methoxyphenylboronic acid (2.02 g, 10.8 mmol, 1.0 eq.), (R)-4-chloro-N-(1-methylpiperidin-3-yl)phthalazin-1-amine (3.00 g, 10.8 mmol, 1.0 eq.), Pd(dppf)Cl2 (790 mg, 1.08 mmol, 0.1 eq.), and NaHCO3 (1.81 g, 21.6 mmol, 2.0 eq.) were added successively to 1,4-dioxane (30 mL) and water (15 mL), and the mixture was heated to 110° C. and reacted for 2 h in nitrogen atmosphere. The reaction solution was cooled to room temperature, and water (60 mL) was added to quench the reaction, followed by extraction with EA (30 mL×2). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (DCM:MeOH=50:1 to 10:1) to give the product (3.50 g, yield: 84.3%).
(R)-4-(4-chloro-2-methoxyphenyl)-N-(1-methylpiperidin-3-yl)phthalazin-1-amine (3.50 g, 9.14 mmol, 1.0 eq.) was dissolved in DCM (30 mL), and the mixture was cooled to 0° C., followed by dropwise addition of BBr3 (4.58 g, 18.3 mmol, 2.0 eq.). The resulting mixture was reacted for 6 h. MeOH was added to quench the reaction, and a saturated aqueous NaHCO3 solution was added to adjust the pH to 8, followed by extraction with DCM (30 mL×2). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (DCM:MeOH=50:1 to 10:1) to give the product (1.44 g, yield: 42.7%).
1HNMR (400 MHz, CDCl3) δ(ppm): 8.26-8.24 (s, 1H), 8.01 (d, J=8.0 Hz, 1H), 7.91-7.82 (m, 2H), 7.55 (d, J=8.4 Hz, 1H), 7.19 (d, J=2.4 Hz, 1H), 7.00-6.98 (m, 1H), 6.29 (s, 1H), 4.71-4.69 (m, 1H), 2.82 (s, 2H), 2.52 (d, J=9.6 Hz, 1H), 2.35 (s, 3H), 2.18-2.10 (m, 2H), 1.89-1.80 (m, 1H), 1.68-1.64 (m, 2H).
Molecular formula: C20H21ClN4O Precise molecular weight: 368.14 LC-MS (Pos, m/z)=369.08[M+H]+.
3-Bromo-5-hydroxypyridine (10.0 g, 57.5 mmol, 1.0 eq.) was dissolved in THF (150 mL), and the mixture was cooled to 0° C., followed by portionwise addition of NaH (60% by mass, 3.45 g, 86.2 mmol, 1.5 eq.). The resulting mixture was stirred for 20 min, and chloromethyl ethyl ether (8.15 g, 86.2 mmol, 1.5 eq.) was added. After 1 h of reaction, a saturated aqueous NH4Cl solution was added to quench the reaction, followed by liquid separation. The aqueous phase was extracted with EA (30 mL×2), and the organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure, and the crude product was separated by silica gel column chromatography (DCM:MeOH=100:1 to 50:1) to give the product (10.8 g, yield: 81.0%).
3-Bromo-5-(ethoxymethoxy)pyridine (10.0 g, 43.1 mmol, 1.0 eq.), cyclopropylboronic acid (4.44 g, 51.7 mmol, 1.2 eq.), Pd(dppf)Cl2 (3.15 g, 4.31 mmol, 0.1 eq.) and NaHCO3 (7.24 g, 86.2 mmol, 2.0 eq.) were added successively to 1,4-dioxane (100 mL) and water (20 mL), and the mixture was heated to 110° C. and reacted for 6 h in nitrogen atmosphere. The reaction solution was cooled to room temperature, and water (50 mL) was added to quench the reaction, followed by extraction with EA (50 mL×3). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (DCM:MeOH=100:1 to 50:1) to give the product (6.20 g, yield: 74.5%).
3-Cyclopropyl-5-(ethoxymethoxy)pyridine (6.00 g, 31.0 mmol, 1.0 eq.) was dissolved in DCM (50 mL), followed by dropwise addition of a solution of hydrogen chloride in 1,4-dioxane (4 mol/L, 15.5 mL, 62.0 mmol, 2.0 eq.). The mixture was stirred at room temperature for 6 h. The pH was adjusted to 8 with a saturated aqueous NaHCO3 solution, followed by extraction with a mixed solvent (DCM:MeOH=10:1) (50 mL×2). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (DCM:MeOH=100:1 to 20:1) to give the product (3.30 g, yield: 78.6%).
3-Hydroxy-5-cyclopropylpyridine (3.20 g, 23.7 mmol, 1.0 eq.) was added to H2O (50 mL), followed by addition of K2CO3 (6.55 g, 47.4 mmol, 2.0 eq.). The mixture was stirred for dissolution, and then 12 (6.01 g, 23.7 mmol, 1.0 eq.) was added. The mixture was reacted at room temperature for 20 h. The pH was adjusted to 7 with dilute hydrochloric acid, followed by extraction with EA (50 mL×3). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (DCM:MeOH=100:1 to 20:1) to give the product (4.70 g, yield: 76.0%).
5-Cyclopropyl-2-iodo-3-hydroxypyridine (4.00 g, 15.3 mmol, 1.0 eq.) was dissolved in THF (80 mL), and the mixture was cooled to 0° C., followed by portionwise addition of NaH (60% by mass, 0.920 g, 23.0 mmol, 1.5 eq.). The resulting mixture was stirred for 30 min, and chloromethyl ethyl ether (2.17 g, 23.0 mmol, 1.5 eq.) was added. The mixture was reacted for 4 h. A saturated aqueous NH4Cl solution was added to quench the reaction, followed by extraction with EA (50 mL×2). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (PE:EA=20:1 to 5:1) to give the product (4.20 g, yield: 85.9%).
5-Cyclopropyl-3-(ethoxymethoxy)-2-iodopyridine (2.00 g, 6.27 mmol, 1.0 eq.) was dissolved in THF (40 mL), and the mixture was cooled to −60° C. in nitrogen atmosphere, followed by dropwise addition of n-butyllithium (1.6 mol/L tetrahydrofuran solution, 4.7 mL, 7.52 mmol, 1.2 eq.). The mixture was stirred for 1 h, and tributyltin chloride (2.45 g, 7.52 mmol, 1.2 eq.) was added. The mixture was reacted for 1 h. A saturated aqueous potassium fluoride solution was added to quench the reaction, followed by extraction with EA (30 mL×3). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (PE:EA=10:1) to give the product (2.40 g, yield: 79.4%).
5-Cyclopropyl-3-(ethoxymethoxy)-2-(tributylstannyl)pyridine (1.05 g, 2.17 mmol, 1.5 eq.), (R)-4-chloro-N-(1-methylpiperidin-3-yl)phthalazin-1-amine (400 mg, 1.45 mmol, 1.0 eq.), tetrakis(triphenylphosphine)palladium (167 mg, 0.145 mmol, 0.1 eq.), cuprous iodide (27.5 mg, 0.145 mmol, 0.1 eq.) and cesium fluoride (440 mg, 2.90 mmol, 2.0 eq.) were added successively to DMF (15 mL), and the mixture was heated to 80° C. and reacted for 2 h in nitrogen atmosphere. The reaction solution was cooled to room temperature, and a saturated aqueous potassium fluoride solution was added to quench the reaction, followed by extraction with EA (30 mL×3). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (DCM:MeOH=20:1 to 10:1) to give the product (530 mg, yield: 84.6%).
(R)-4-(5-cyclopropyl-3-(ethoxymethoxy)pyridin-2-yl)-N-(1-methylpiperidin-3-yl)phthalazin-1-amine (530 mg, 1.22 mmol, 1.0 eq.) was dissolved in DCM (8 mL), followed by dropwise addition of a solution of hydrogen chloride in 1,4-dioxane (4 mol/L, 1.53 mL, 6.10 mmol, 5.0 eq.). The mixture was stirred at room temperature for 1 h. The pH was adjusted to 9 with a saturated aqueous Na2CO3 solution, followed by extraction with DCM (20 mL×3). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was separated by preparative thin-layer chromatography (DCM:MeOH=50:1) to give the product (432 mg, yield: 94.1%).
1HNMR (400 MHz, DMSO-d6) δ (ppm): 14.55 (s, 1H), 9.73-9.71 (m, 1H), 8.60 (s, 1H), 8.11 (d, J=2.0 Hz, 1H), 7.97-7.94 (m, 2H), 7.75 (s, 1H), 7.01 (d, J=2.0 Hz, 1H), 4.59 (s, 1H), 3.31 (d, J=8.8 Hz, 1H), 2.96 (d, J=11.2 Hz, 1H), 2.48 (s, 4H), 2.00-1.97 (m, 2H), 1.87 (s, 1H), 1.75-1.69 (m, 2H), 1.06-1.03 (m, 2H), 0.84-0.82 (m, 1H).
Molecular formula: C22H25N5O Precise molecular weight: 375.21 LC-MS (Pos, m/z)=376.11[M+H]+.
(R)-4-chloro-N-(piperidin-3-yl)phthalazin-1-amine (300 mg, 1.14 mmol, 1.0 eq.) and tetrahydro-4H-pyran-4-one (171 mg, 1.71 mmol, 1.5 eq.) were dissolved in MeOH (5 mL), and the mixture was stirred at room temperature for 1 h. NaBH3CN (107 mg, 3.36 mmol, 1.5 eq.) was added, and the resulting mixture was reacted for 25 h. A saturated aqueous NaHCO3 solution was added to quench the reaction, followed by extraction with DCM (20 mL×2). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (DCM:MeOH=50:1 to 20:1) to give the product (312 mg, yield: 78.8%).
3-(Ethoxymethoxy)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzonitrile (324 mg, 1.07 mmol, 1.2 eq.), (R)-4-chloro-N-(1-(tetrahydro-2H-pyran-4-yl)piperidin-3-yl)phthalazin-1-amine (310 mg, 0.894 mmol, 1.0 eq.), Pd(dppf)Cl2 (65.4 mg, 0.0894 mmol, 0.1 eq.) and NaHCO3 (150 mg, 1.79 mmol, 2.0 eq.) were added successively to 1,4-dioxane (6 mL) and water (3 mL), and the mixture was heated to 110° C. and reacted for 2 h in nitrogen atmosphere. The reaction solution was cooled to room temperature, and water (30 mL) was added to quench the reaction, followed by extraction with EA (30 mL×2). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (DCM:MeOH=50:1 to 20:1) to give the product (290 mg, yield: 67.7%).
(R)-3-(ethoxymethoxy)-4-(4-((1-(tetrahydro-2H-pyran-4-yl)piperidin-3-yl)amino)phthalazin-1-yl)benzonitrile (290 mg, 0.595 mmol, 1.0 eq.) was dissolved in DCM (3 mL), followed by dropwise addition of a solution of hydrogen chloride in 1,4-dioxane (4 mol/L, 0.75 mL, 2.98 mmol, 5.0 eq.). The mixture was stirred at room temperature for 2 h. The pH was adjusted to 9 with a saturated aqueous Na2CO3 solution, followed by extraction with DCM (20 mL×3). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was slurried with a mixed solvent (PE:EA=10:1) to give the product (165 mg, yield: 64.6%).
1HNMR (400 MHz, DMSO-d6) δ (ppm): 10.61 (s, 1H), 8.39 (s, 1H), 7.86 (s, 1H), 7.79 (s, 1H), 7.49 (d, J=6.8 Hz, 1H), 7.41 (s, 2H), 7.31 (s, 1H), 7.15 (s, 1H), 4.39 (s, 1H), 3.88 (d, J=9.2 Hz, 2H), 3.30-3.24 (m, 4H), 2.84 (d, J=8.0 Hz, 1H), 2.17 (d, J=6.8 Hz, 2H), 2.01 (s, 1H), 1.75 (s, 1H), 1.66 (s, 2H), 1.52-1.49 (m, 4H).
Molecular formula: C25H27N5O2 Precise molecular weight: 429.22 LC-MS (Pos, m/z)=430.17[M+H]+.
(R)-4-chloro-N-(piperidin-3-yl)phthalazin-1-amine (300 mg, 1.14 mmol, 1.0 eq.) was dissolved in MeOH (5 mL), followed by addition of acetone (99.3 mg, 1.71 mmol, 1.5 eq.). The mixture was stirred at room temperature for 2 h. NaBH3CN (107 mg, 3.36 mmol, 1.5 eq.) was added, and the resulting mixture was reacted for 25 h. A saturated aqueous NaHCO3 solution was added to quench the reaction, followed by extraction with DCM (20 mL×2). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (DCM:MeOH=50:1 to 20:1) to give the product (280 mg, yield: 80.4%).
3-(Ethoxymethoxy)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzonitrile (334 mg, 1.10 mmol, 1.2 eq.), (R)-4-chloro-N-(1-isopropylpiperidin-3-yl)phthalazin-1-amine (280 mg, 0.919 mmol, 1.0 eq.), Pd(dppf)Cl2 (67.2 mg, 0.0917 mmol, 0.1 eq.) and NaHCO3 (155 mg, 1.84 mmol, 2.0 eq.) were added successively to 1,4-dioxane (6 mL) and water (3 mL), and the mixture was heated to 110° C. and reacted for 2 h in nitrogen atmosphere. The reaction solution was cooled to room temperature, and water (30 mL) was added to quench the reaction, followed by extraction with EA (30 mL×2). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (DCM:MeOH=50:1 to 10:1) to give the product (295 mg, yield: 65.5%).
(R)-3-(ethoxymethoxy)-4-(4-((1-isopropylpiperidin-3-yl)amino)phthalazin-1-yl)benzonitrile (295 mg, 0.662 mmol, 1.0 eq.) was dissolved in DCM (3 mL), followed by dropwise addition of a solution of hydrogen chloride in 1,4-dioxane (4 mol/L, 0.83 mL, 3.31 mmol, 5.0 eq.). The mixture was stirred at room temperature for 2 h. The pH was adjusted to 9 with a saturated aqueous Na2CO3 solution, followed by extraction with DCM (20 mL×3). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was separated by preparative thin-layer chromatography (DCM:MeOH=10:1) to give the product (152 mg, yield: 59.2%).
1HNMR (400 MHz, DMSO-d6) δ (ppm): 10.51 (s, 1H), 8.39 (d, J=8.4 Hz, 1H), 7.88-7.84 (m, 1H), 7.80-7.77 (m, 1H), 7.50 (d, J=7.6 Hz, 1H), 7.42 (d, J=7.6 Hz, 2H), 7.32 (d, J=1.2 Hz, 1H), 7.15 (d, J=8.0 Hz, 1H), 4.41-4.39 (m, 1H), 3.11 (d, J=8.0 Hz, 1H), 2.76 (s, 2H), 2.17-2.15 (m, 2H), 2.01-2.99 (m, 1H), 1.77-1.74 (m, 1H), 1.57-1.46 (m, 2H), 1.01-0.98 (m, 6H).
Molecular formula: C23H25N5O Precise molecular weight: 387.21 LC-MS (Pos, m/z)=388.14[M+H]+.
3-Bromo-2-methoxy-6-methylpyridine (6.00 g, 29.7 mmol, 1.0 eq.) and triisopropyl borate (11.3 g, 59.4 mmol, 2.0 eq.) were dissolved in THF (60 mL), followed by purge with nitrogen for protection. The mixture was cooled to −70° C., followed by dropwise addition of n-butyllithium (23.8 mL, 59.4 mmol, 2.0 eq.). The mixture was reacted for 2 h. Dilute hydrochloric acid was added to quench the reaction, and the pH was adjusted to 3. The mixture was stirred at room temperature for 30 min and then extracted with a mixed solvent of DCM:MeOH (10:1) (50 mL×5). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure and slurried with petroleum ether to give (2-methoxy-6-methylpyridin-3-yl)boronic acid (4.20 g, yield: 84.7%).
(2-Methoxy-6-methylpyridin-3-yl)boronic acid (3.0 g, 18.0 mmol, 1.0 eq.), 1,4-dichlorophthalazine (4.30 g, 21.6 mmol, 1.2 eq.), Pd(dppf)Cl2 (659 mg, 0.900 mmol, 0.05 eq.) and NaHCO3 (3.02 g, 36.0 mmol, 2.0 eq.) were added successively to 1,4-dioxane (30 mL), and H2O (15 mL) was added, followed by purge with nitrogen for protection. The oil bath was heated to 110° C. and the mixture was reacted for 2 h. The reaction solution was cooled down, and water was added to quench the reaction, followed by extraction with EA (40 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (PE:EA=10:1) to give the product (2.20 g, yield: 42.9%).
1-Chloro-4-(2-methoxy-6-methylpyridin-3-yl)phthalazine (2.00 g, 7.00 mmol, 1.0 eq.), tert-butyl (R)-3-aminopiperidine-1-carboxylate (1.54 g, 7.70 mmol, 1.1 eq.), Pd2(dba)3 (641 mg, 0.700 mmol, 0.1 eq.), BINAP (871 mg, 1.40 mmol, 0.2 eq.) and Cs2CO3 (4.56 g, 14.0 mmol, 2.0 eq.) were added successively to toluene (40 mL), followed by purge with nitrogen for protection. The oil bath was heated to 90° C. and the mixture was reacted for 20 h. The reaction solution was cooled down, and water was added to quench the reaction, followed by extraction with EA. The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (DCM:MeOH=100:1 to 40:1) to give the product (1.50 g, yield: 47.7%).
Tert-butyl (R)-3-((4-(2-methoxy-6-methylpyridin-3-yl)phthalazin-1-yl)amino)piperidine-1-carboxylate (1.00 g, 2.22 mmol, 1.0 eq.) was dissolved in DCM (20 mL), and the mixture was cooled to −10° C., followed by dropwise addition of BBr3 (2.78 g, 11.1 mmol, 5.0 eq.). The mixture was reacted for 4 h. MeOH was added to quench the reaction, and the reaction solution was concentrated. The crude product was dissolved in H2O, followed by extraction with EA. A saturated aqueous NaHCO3 solution was added to the aqueous phase to adjust the pH to 8, and DCM (20 mL×3) was added for extraction. The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to give the product (480 mg, yield: 64.3%)
(R)-6-methyl-3-(4-(piperidin-3-ylamino)phthalazin-1-yl)pyridin-2-ol (400 mg, 1.19 mmol, 1.0 eq.) was dissolved in MeOH (10 mL), followed by addition of an aqueous formaldehyde solution (37%, 143 mg, 3.57 mmol, 3.0 eq.). The mixture was stirred at room temperature for 30 min. NaBH3CN (224 mg, 3.57 mmol, 3.0 eq.) was then added and the mixture was reacted for 2 h. Water was added to quench the reaction, followed by extraction with EA (30 mL×3). The aqueous phase was concentrated under reduced pressure, and the crude product was separated by reversed phase preparative column chromatography (acetonitrile: 0.1% ammonia solution=4:1) to give the product (180 mg, yield: 43.2%).
1HNMR (300 MHz, DMSO-d6) δ (ppm): 11.95 (s, 1H), 8.50 (s, 1H), 7.89-7.77 (m, 2H), 7.60-7.52 (m, 3H), 6.19 (d, J=6.9 Hz, 1H), 4.74 (s, 1H), 3.63-3.59 (m, 2H), 3.17 (s, 1H), 2.78 (s, 4H), 2.30 (s, 3H), 2.08 (s, 1H), 1.84 (s, 3H).
Molecular formula: C20H23N5O Molecular weight: 349.44 LC-MS (Pos, m/z)=350.28[M+H]+.
1,4-Dichlorophthalazine (1.0 g, 5.02 mmol, 1.0 eq.), 3-(ethoxymethoxy)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzonitrile (1.82 g, 6.02 mmol, 1.2 eq.), sodium bicarbonate (844.1 mg, 10.04 mmol, 2.0 eq.) and Pd(dppf)Cl2 (183.8 mg, 0.25 mmol, 0.05 eq.) were added to a mixed solution of 1,4-dioxane (15.0 mL) and water (7.0 mL), and the mixture was reacted at 95° C. for 12 h in nitrogen atmosphere. After the reaction was completed as monitored by TLC, water (100.0 mL) was added, followed by extraction with ethyl acetate (100.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=20:1 to 2:1) to give the product (680.4 mg, yield: 40.0%).
4-(4-Chlorophthalazin-1-yl)-3-(ethoxymethoxy)benzonitrile (200.0 mg, 0.58 mmol, 1.0 eq.), (cis)-3-amino-1-methylcyclobutan-1-ol hydrochloride (161.9 mg, 1.17 mmol, 2.0 eq.) and N,N-diisopropylethylamine (303.9 mg, 2.35 mmol, 4.0 eq.) were added to N,N-dimethylacetamide (6.0 mL), and the mixture was reacted at 110° C. for 5 h. After the reaction was completed as monitored by TLC, water (50.0 mL) was added, followed by extraction with ethyl acetate (100.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to give the crude product (315.0 mg, used directly in the next step).
3-(Ethoxymethoxy)-4-(4-(((cis)-3-hydroxy-3-methylcyclobutyl)amino)phthalazin-1-yl)benzonitrile (315.0 mg crude product, 0.58 mmol, 1.0 eq.) was added to dichloromethane (2.0 mL), followed by dropwise addition of a hydrogen chloride-1,4-dioxane solution (4.0 mol/L, 3.0 mL). The mixture was reacted at room temperature for 2 h. After the reaction was completed as monitored by TLC, the pH of the system was adjusted to 7-8 with a saturated aqueous sodium bicarbonate solution, followed by extraction with dichloromethane (100.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by preparative thin-layer chromatography (dichloromethane:methanol=10:1) to give the product (20.0 mg, yield: 9.8%).
1HNMR (400 MHz, DMSO-d6) δ (ppm): 10.47 (s, 1H), 8.44-8.42 (d, J=8 Hz, 1H), 7.86 (t, J=8 Hz, 1H), 7.78 (t, J=8 Hz, 1H), 7.67-7.65 (d, J=8 Hz, 1H), 7.50-7.48 (d, J=8 Hz, 1H), 7.42-7.40 (d, J=8 Hz, 2H), 7.33-7.32 (d, J=4 Hz, 1H), 5.02 (s, 1H), 4.27-4.22 (m, 1H), 2.50-2.46 (m, 2H), 2.20-2.15 (m, 2H), 1.34 (s, 3H).
Molecular formula: C20H18N4O2 Precise molecular weight: 346.14 LC-MS (m/z)=347.10[M+H]+.
4-Bromo-3-methylbenzonitrile (1.0 g, 5.10 mmol, 1.0 eq.), 4,4,5,5-tetramethyl-1,3,2-dioxaborolane (1.94 g, 7.65 mmol, 1.5 eq.), potassium acetate (1.0 g, 10.20 mmol, 2.0 eq.) and Pd(dppf)Cl2 (186.5 mg, 0.25 mmol, 0.05 eq.) were added to 1,4-dioxane (20.0 mL), and the mixture was reacted at 100° C. for 4 h in nitrogen atmosphere. After the reaction was completed as monitored by TLC, water (100.0 mL) was added, followed by extraction with ethyl acetate (100.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (specification of silica gel: 100-200 mesh, petroleum ether:ethyl acetate=10:1) to give the product (1.0 g, yield: 80.7%).
(R)-4-chloro-N-(1-methylpiperidin-3-yl)phthalazin-1-amine (100.0 mg, 0.36 mmol, 1.0 eq.), 3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzonitrile (131.7 mg, 0.54 mmol, 1.5 eq.), sodium bicarbonate (60.6 mg, 0.72 mmol, 2.0 eq.) and Pd(dppf)Cl2 (13.2 mg, 0.01 mmol, 0.05 eq.) were added to a mixed solution of 1,4-dioxane (10.0 mL) and water (5.0 mL), and the mixture was reacted at 110° C. for 3 h in nitrogen atmosphere. After the reaction was completed as monitored by TLC, water (100.0 mL) was added, followed by extraction with ethyl acetate (100.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by preparative thin-layer chromatography (dichloromethane:methanol=10:1) to give the product (40.0 mg, yield: 31.0%).
1HNMR (400 MHz, DMSO-d6) δ (ppm): 8.47-8.45 (d, J=8 Hz, 1H), 7.91-7.87 (m, 2H), 7.83-7.78 (m, 2H), 7.51-7.49 (d, J=8 Hz, 1H), 7.26-7.25 (d, J=4 Hz, 1H), 7.21-7.19 (d, J=8 Hz, 1H), 4.44-4.42 (m, 1H), 3.09-3.07 (d, J=8 Hz, 1H), 2.72-2.70 (d, J=8 Hz, 1H), 2.21 (s, 3H), 2.06 (s, 3H), 2.01-1.89 (m, 3H), 1.77-1.40 (m, 3H).
Molecular formula: C22H23N5 Precise molecular weight: 357.20 LC-MS (m/z)=358.12[M+H]+.
1,4-Dichlorophthalazine (1.92 g, 9.68 mmol, 1.0 eq.), (cis)-3-amino-1-methylcyclobutan-1-ol hydrochloride (1.6 g, 11.62 mmol, 1.2 eq.) and N,N-diisopropylethylamine (1.87 g, 14.53 mmol, 1.5 eq.) were added to N,N-dimethylacetamide (20.0 mL), and the mixture was reacted at 120° C. for 12 h. After the reaction was completed as monitored by TLC, water (100.0 mL) was added, followed by extraction with ethyl acetate (100.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (specification of silica gel: 100-200 mesh, dichloromethane:methanol=100:1 to 40:1) to give the product (900.0 mg, yield: 36.0%).
(Cis)-3-((4-chlorophthalazin-1-yl)amino)-1-methylcyclobutan-1-ol (250.0 mg, 0.94 mmol, 1.0 eq.), (2-(ethoxymethoxy)-4-(trifluoromethyl)phenyl)boronic acid (350.3 mg, 1.32 mmol, 1.4 eq.), sodium bicarbonate (159.1 mg, 1.89 mmol, 2.0 eq.) and Pd(dppf)Cl2 (34.6 mg, 0.04 mmol, 0.05 eq.) were added to a mixed solution of 1,4-dioxane (10.0 mL) and water (5.0 mL), and the mixture was reacted at 110° C. for 2 h in nitrogen atmosphere. After the reaction was completed as monitored by TLC, water (100.0 mL) was added, followed by extraction with ethyl acetate (100.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (dichloromethane:methanol=80:1 to 30:1) to give the product (320.0 mg, yield: 75.0%).
(Cis)-3-((4-(2-(ethoxymethoxy)-4-(trifluoromethyl)phenyl)phthalazin-1-yl)amino)-1-methylcyclobutan-1-ol (310.0 mg, 0.69 mmol, 1.0 eq.) was added to dichloromethane (2.0 mL), followed by dropwise addition of a hydrogen chloride-1,4-dioxane solution (4.0 mol/L, 4.0 mL). The mixture was reacted at room temperature for 1 h. After the reaction was completed as monitored by TLC, the pH of the system was adjusted to 7-8 with a saturated aqueous sodium bicarbonate solution, followed by extraction with dichloromethane (50.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (specification of silica gel: 100-200 mesh, dichloromethane:methanol=40:1 to 30:1) to give the product (180.0 mg, yield: 66.9%).
1HNMR (400 MHz, DMSO-d6) δ (ppm): 10.37 (s, 1H), 8.44-8.42 (d, J=8 Hz, 1H), 7.86 (t, J=8 Hz, 1H), 7.77 (t, J=8 Hz, 1H), 7.64-7.62 (d, J=8 Hz, 1H), 7.52-7.50 (d, J=8 Hz, 1H), 7.45-7.43 (d, J=8 Hz, 1H), 7.31-7.28 (m, 2H), 5.02 (s, 1H), 4.27-4.22 (m, 1H), 2.48-2.47 (m, 2H), 2.20-2.15 (m, 2H), 1.34 (s, 3H).
Molecular formula: C20H18F3N3O2 Precise molecular weight: 389.14 LC-MS (m/z)=390.06[M+H]+.
4-(4-Chlorophthalazin-1-yl)-3-(ethoxymethoxy)benzonitrile (310.0 mg, 0.91 mmol, 1.0 eq.), 1-(tert-butyl) 2-ethyl (2S,5R)-5-aminopiperidine-1,2-dicarboxylate (298.1 mg, 1.09 mmol, 1.2 eq.), cesium carbonate (594.2 mg, 1.82 mmol, 2.0 eq.), Pd2(dba)3 (83.5 mg, 0.09 mmol, 0.1 eq.) and 1,1′-binaphthyl-2,2′-bisdiphenylphosphine (113.5 mg, 0.18 mmol, 0.2 eq.) were added to 1,4-dioxane (15.0 mL), and the mixture was reacted at 90° C. for 12 h in nitrogen atmosphere. After the reaction was completed as monitored by TLC, water (100.0 mL) was added, followed by extraction with ethyl acetate (100.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (specification of silica gel: 100-200 mesh, dichloromethane:methanol=100:1 to 60:1) to give the product (262.5 mg, yield: 50%).
1-(Tert-butyl) 2-ethyl (2S,5R)-5-((4-(4-cyano-2-(ethoxymethoxy)phenyl)phthalazin-1-yl)amino)piperidine-1,2-dicarboxylate (260.0 mg, 0.45 mmol, 1.0 eq.) was added to dichloromethane (2.0 mL), followed by dropwise addition of a hydrogen chloride-1,4-dioxane solution (4.0 mol/L, 4.0 mL). The mixture was reacted at room temperature for 2 h. After the reaction was completed as monitored by TLC, the pH of the system was adjusted to 7-8 with a saturated aqueous sodium bicarbonate solution, followed by extraction with dichloromethane (100.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to give the product (159.4 mg, yield: 90%).
Ethyl (2S,5R)-5-((4-(4-cyano-2-hydroxyphenyl)phthalazin-1-yl)amino)piperidine-2-carboxylate (169.0 mg, 0.40 mmol, 1.0 eq.), an aqueous formaldehyde solution (37%) (46.0 mg, 0.56 mmol, 1.4 eq.) and acetic acid (0.5 mL) were added to methanol (10.0 mL), and the mixture was stirred at room temperature for 0.5 h, followed by addition of sodium cyanoborohydride (38.0 mg, 0.60 mmol, 1.5 eq.). The resulting mixture was reacted at room temperature for 2 h. After the reaction was completed as monitored by TLC, the system was concentrated under reduced pressure, followed by addition of a saturated aqueous sodium bicarbonate solution (100.0 mL). The mixture was stirred for 0.5 h and extracted with dichloromethane (100.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by preparative thin-layer chromatography (dichloromethane:methanol=10:1) to give the product (139.4 mg, yield: 80.0%).
Ethyl (2S,5R)-5-((4-(4-cyano-2-hydroxyphenyl)phthalazin-1-yl)amino)-1-methylpiperidine-2-carboxylate (139.4 mg, 0.32 mmol, 1.0 eq.) and lithium hydroxide (135.5 mg, 3.23 mmol, 10.0 eq.) were added to a mixed solution of methanol (10.0 mL) and water (4.0 mL), and the mixture was stirred at room temperature for 72 h. After the reaction was completed as monitored by TLC, the system was concentrated under reduced pressure and purified by reversed phase column chromatography (acetonitrile:0.1% ammonia solution=4:6) to give the ammonium salt of the product (84.0 mg, yield: 64.4%).
1HNMR (400 MHz, CD3OD) δ (ppm): 8.39-8.37 (d, J=8 Hz, 1H), 8.01-7.97 (m, 1H), 7.90 (t, J=8 Hz, 1H), 7.67-7.65 (d, J=8 Hz, 1H), 7.56-7.54 (d, J=8 Hz, 1H), 7.41-7.39 (m, 1H), 7.32 (s, 1H), 4.74-4.68 (m, 1H), 3.99-3.96 (m, 1H), 3.54-3.51 (m, 1H), 3.02-2.93 (m, 4H), 2.52-2.33 (m, 2H), 2.06-1.91 (m, 2H).
Molecular formula: C22H21N5O3 Precise molecular weight: 403.16 LC-MS (m/z)=404.13[M+H]+.
(R)-4-chloro-N-(piperidin-3-yl)phthalazin-1-amine (300 mg, 1.14 mmol, 1.0 eq.), 1-chloro-2-methyl-2-propanol (248 mg, 2.28 mmol, 2.0 eq.) and K2CO3 (315 mg, 2.28 mmol, 2.0 eq.) were added to DMF (5 mL), and the mixture was heated to 90° C. and reacted for 24 h. The reaction solution was cooled to room temperature, and water (30 mL) was added to quench the reaction, followed by extraction with EA (30 mL). The organic phase was washed with water (20 mL×3), dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure and separated by silica gel column chromatography (DCM:MeOH=100:1 to 20:1) to give the product (240 mg, yield: 62.8%).
3-(Ethoxymethoxy)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzonitrile (261 mg, 0.860 mmol, 1.2 eq.), (R)-2-(3-((4-chlorophthalazin-1-yl)amino)piperidin-1-yl)-2-methyl-2-propanol (240 mg, 0.717 mmol, 1.0 eq.), Pd(dppf)Cl2 (52.5 mg, 0.0717 mmol, 0.1 eq.) and NaHCO3 (120 mg, 1.43 mmol, 2.0 eq.) were added successively to 1,4-dioxane (6 mL), followed by addition of H2O (3 mL). The mixture was heated to 110° C. and reacted for 2 h in nitrogen atmosphere. The reaction solution was cooled to room temperature, and water (30 mL) was added to quench the reaction, followed by extraction with EA (30 mL×2). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure and separated by silica gel column chromatography (DCM:MeOH=50:1 to 20:1) to give the product (285 mg, yield: 83.6%).
(R)-3-(ethoxymethoxy)-4-(4-((1-(2-hydroxy-2-methylpropyl)piperidin-3-yl)amino)phthalazin-1-yl)benzonitrile (285 mg, 0.599 mmol, 1.0 eq.) was dissolved in DCM (3 mL), followed by dropwise addition of a solution of hydrogen chloride in 1,4-dioxane (4 mol/L, 0.75 mL, 3.00 mmol, 5.0 eq.). The mixture was stirred at room temperature for 1 h. The pH was adjusted to 9 with a saturated aqueous Na2CO3 solution, followed by extraction with DCM (20 mL×3). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure and separated by silica gel column chromatography (DCM:MeOH=50:1 to 10:1) to give the product (146 mg, yield: 58.4%).
1HNMR (400 MHz, DMSO-d6) δ (ppm): 10.53 (s, 1H), 8.40 (d, J=8.0 Hz, 1H), 7.89-7.85 (m, 1H), 7.81-7.77 (m, 1H), 7.50 (d, J=8.0 Hz, 1H), 7.43-7.40 (m, 2H), 7.37 (d, J=0.8 Hz, 1H), 7.18 (s, 1H), 4.51 (s, 1H), 4.13 (s, 1H), 3.20 (s, 1H), 2.90 (s, 1H), 2.28 (s, 3H), 1.96 (s, 1H), 1.75-1.51 (m, 3H), 1.14 (s, 6H).
Molecular formula: C24H27N5O2 Precise molecular weight: 417.22 LC-MS (Pos, m/z)=418.13 [M+H]+.
(R)-4-chloro-N-(piperidin-3-yl)phthalazin-1-amine (300 mg, 1.14 mmol, 1.0 eq.), 2-bromoethanol (712 mg, 5.70 mmol, 5.0 eq.) and triethylamine (577 mg, 5.70 mmol, 5.0 eq.) were added to DCM (10 mL), and the mixture was reacted at room temperature for 25 h. Water (20 mL) was added to quench the reaction, followed by extraction with DCM (20 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure and separated by silica gel column chromatography (DCM:MeOH=50:1 to 15:1) to give the product (212 mg, yield: 60.5%).
3-(Ethoxymethoxy)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzonitrile (249 mg, 0.822 mmol, 1.2 eq.), (R)-2-(3-((4-chlorophthalazin-1-yl)amino)piperidin-1-yl)-1-ethanol (210 mg, 0.685 mmol, 1.0 eq.), Pd(dppf)Cl2 (50.1 mg, 0.0685 mmol, 0.1 eq.) and NaHCO3 (115 mg, 1.37 mmol, 2.0 eq.) were added successively to 1,4-dioxane (6 mL), followed by addition of H2O (3 mL). The mixture was heated to 110° C. and reacted for 2 h in nitrogen atmosphere. The reaction solution was cooled to room temperature, and water (30 mL) was added to quench the reaction, followed by extraction with EA (30 mL×2). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure and separated by silica gel column chromatography (DCM:MeOH=50:1 to 10:1) to give the product (180 mg, yield: 58.8%).
(R)-3-(ethoxymethoxy)-4-(4-((1-(2-hydroxyethyl)piperidin-3-yl)amino)phthalazin-1-yl)benzonitrile (180 mg, 0.402 mmol, 1.0 eq.) was dissolved in DCM (3 mL), followed by dropwise addition of a solution of hydrogen chloride in 1,4-dioxane (4 mol/L, 0.50 mL, 2.01 mmol, 5.0 eq.). The mixture was stirred at room temperature for 1 h. The pH was adjusted to 9 with a saturated aqueous Na2CO3 solution, followed by extraction with DCM (20 mL×3). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure and separated by reversed phase column chromatography (acetonitrile:0.1% ammonia solution=2:5) to give the product (72.0 mg, yield: 46.0%).
1HNMR (400 MHz, DMSO-d6) δ (ppm): 10.50 (s, 1H), 8.40 (d, J=8.0 Hz, 1H), 7.88-7.84 (m, 1H), 7.80-7.77 (m, 1H), 7.50 (d, J=8.0 Hz, 1H), 7.43-7.42 (m, 2H), 7.33 (s, 1H), 7.15 (d, J=7.6 Hz, 1H), 4.43 (s, 2H), 3.54-3.51 (m, 2H), 3.17-3.14 (m, 1H), 2.81 (d, J=8.0 Hz, 1H), 2.47-2.44 (m, 2H), 2.11-2.03 (m, 2H), 1.98 (d, J=8.0 Hz, 1H), 1.75 (d, J=12.0 Hz, 1H), 1.61-1.52 (m, 2H).
Molecular formula: C22H23N5O2 Precise molecular weight: 389.19 LC-MS (Pos, m/z)=390.12[M+H]+.
(S)-1-methylpiperidine-2-carboxylic acid (180 mg, 1.26 mmol, 1.0 eq.) was dissolved in DCM (5 mL), followed by addition of N-methylmorpholine (191 mg, 1.89 mmol, 1.5 eq.) and isopropyl chloroformate (232 mg, 1.89 mmol, 1.5 eq.). The mixture was stirred at room temperature for 2 h, followed by addition of 4-chlorophthalazin-1-amine (226 mg, 1.26 mmol, 1.0 eq.). The resulting mixture was stirred at room temperature for 20 h. H2O (20 mL) was added, followed by extraction with DCM (20 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure and separated by silica gel column chromatography (DCM:MeOH=100:1 to 20:1) to give the product (160 mg, yield: 41.8%).
3-(Ethoxymethoxy)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzonitrile (191 mg, 0.630 mmol, 1.2 eq.), (S)—N-(4-chlorophthalazin-1-yl)-1-methylpiperidine-2-carboxamide (160 mg, 0.525 mmol, 1.0 eq.), Pd(dppf)Cl2 (38.4 mg, 0.0525 mmol, 0.1 eq.) and NaHCO3 (88.2 mg, 1.05 mmol, 2.0 eq.) were added successively to 1,4-dioxane (6 mL), followed by addition of H2O (3 mL). The mixture was heated to 110° C. and reacted for 2 h in nitrogen atmosphere. The reaction solution was cooled to room temperature, and water (20 mL) was added to quench the reaction, followed by extraction with EA (20 mL×2). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure and separated by silica gel column chromatography (DCM:MeOH=100:1 to 20:1) to give the product (80.0 mg, yield: 34.2%).
(S)—N-(4-(4-cyano-2-(ethoxymethoxy)phenyl)phthalazin-1-yl)-1-methylpiperidine-2-carboxamide (80.0 mg, 0.182 mmol, 1.0 eq.) was dissolved in DCM (2 mL), followed by dropwise addition of a solution of hydrogen chloride in 1,4-dioxane (4 mol/L, 0.23 mL, 0.90 mmol, 5.0 eq.). The mixture was stirred at room temperature for 1 h. H2O (5 mL) was added, followed by washing with DCM (5 mL). The aqueous phase was adjusted to pH of 9 with a saturated aqueous Na2CO3 solution and extracted with a mixed solvent (DCM:MeOH=10:1) (10 mL×5). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure and separated by preparative thin-layer chromatography (DCM:MeOH=10:1) to give the product (38.2 mg, yield: 54.9%).
1HNMR (400 MHz, DMSO-d6) δ (ppm): 8.39 (s, 1H), 8.07-7.98 (m, 2H), 7.76 (d, J=8.0 Hz, 1H), 7.58 (d, J=8.0 Hz, 1H), 7.42-7.40 (m, 1H), 7.33 (d, J=1.2 Hz, 1H), 3.25-3.22 (m, 2H), 2.63 (s, 3H), 2.53 (s, 1H), 2.28 (d, J=11.2 Hz, 1H), 1.96-1.89 (m, 2H), 1.87-1.77 (m, 2H), 1.60-1.53 (m, 1H).
Molecular formula: C22H21N5O2 Precise molecular weight: 387.17 LC-MS (Pos, m/z)=388.13[M+H]+.
3-Methoxy-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzonitrile (1.26 g, 4.86 mmol, 1.0 eq.), 4,7-dichlorothieno[2,3-d]pyridazine (997 mg, 4.86 mmol, 1.0 eq.), Pd(dppf)Cl2 (178 mg, 0.243 mmol, 0.05 eq.) and NaHCO3 (817 g, 9.72 mmol, 2.0 eq.) were added successively to 1,4-dioxane (16 mL), followed by addition of H2O (8 mL). The mixture was heated to 110° C. and reacted for 2 h in nitrogen atmosphere. Water (30 mL) was added, followed by extraction with EA (40 mL×2). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified and separated by silica gel column chromatography (PE:EA=5:1 to 1:1) to give the product (500 mg, yield: 34.1%).
The mixture (500 g, 1.16 mmol, 1.0 eq.) of 4-(7-chlorothieno[2,3-d]pyridazin-4-yl)-3-methoxybenzonitrile and 4-(4-chlorothieno[2,3-d]pyridazin-7-yl)-3-methoxybenzonitrile, tert-butyl (R)-3-aminopiperidine-1-carboxylate (499 mg, 2.49 mmol, 1.5 eq.), Pd2(dba)3 (152 mg, 0.166 mmol, 0.1 eq.), BINAP (207 mg, 0.332 mmol, 0.2 eq.) and Cs2CO3 (1.05 g, 3.22 mmol, 2.0 eq.) were added successively to 1,4-dioxane (20 mL), and the mixture was heated to 90° C. and reacted for 20 h in nitrogen atmosphere. The reaction solution was cooled to room temperature, and water (40 mL) was added to quench the reaction, followed by extraction with EA (30 mL×2). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure and purified and separated by silica gel column chromatography (DCM:MeOH=100:1 to 20:1) to give the product (610 mg, yield: 79.1%).
The mixture (600 mg, 1.29 mmol, 1.0 eq.) of tert-butyl (R)-3-((4-(4-cyano-2-methoxyphenyl)thieno[2,3-d]pyridazin-7-yl)amino)piperidine-1-carboxylate and tert-butyl (R)-3-((7-(4-cyano-2-methoxyphenyl)thieno[2,3-d]pyridazin-4-yl)amino)piperidine-1-carboxylate was dissolved in DCM (15 mL), and the mixture was cooled to −10° C., followed by dropwise addition of BBr3 (1.29 g, 5.16 mmol, 4.0 eq.). The resulting mixture was reacted for 5 h. MeOH (5 mL) was added to quench the reaction, and the pH was adjusted to 8 with a saturated aqueous NaHCO3 solution, followed by extraction with DCM (30 mL×2). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to give the product (280 mg, yield: 61.8%).
The mixture (200 mg, 0.596 mmol, 1.0 eq.) of (R)-3-hydroxy-4-(7-(piperidin-3-ylamino)thieno[2,3-d]pyridazin-4-yl)benzonitrile and (R)-3-hydroxy-4-(4-(piperidin-3-ylamino)thieno[2,3-d]pyridazin-7-yl)benzonitrile was dissolved in MeOH (4 mL), followed by addition of an aqueous formaldehyde solution (37%) (139 mg, 1.71 mmol, 3.0 eq.). The mixture was stirred at room temperature for 30 min. NaBH3CN (107 mg, 1.71 mmol, 3.0 eq.) was then added and the mixture was reacted for 1 h. A saturated aqueous NaHCO3 solution was added to quench the reaction, followed by extraction with DCM (20 mL×3). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was separated by preparative thin-layer chromatography (DCM:MeOH=10:1) to give (R)-3-hydroxy-4-(7-((1-methylpiperidin-3-yl)amino)thieno[2,3-d]pyridazin-4-yl)benzonitrile (22.0 mg, yield: 10.6%) and (R)-3-hydroxy-4-(4-((1-methylpiperidin-3-yl)amino)thieno[2,3-d]pyridazin-7-yl)benzonitrile (23.0 mg, yield: 11.1%).
1HNMR (400 MHz, DMSO-d6) δ (ppm): 8.11 (d, J=5.2 Hz, 1H), 7.71 (d, J=8.4 Hz, 1H), 7.41-7.38 (m, 3H), 7.19 (d, J=7.6 Hz, 1H), 4.41-4.39 (m, 1H), 3.05 (d, J=7.6 Hz, 1H), 2.70 (d, J=10.8 Hz, 1H), 2.21 (s, 3H), 1.97-1.92 (m, 3H), 1.75-1.71 (m, 1H), 1.64-1.55 (m, 1H), 1.49-1.35 (m, 1H).
Molecular formula: C19H19N5OS Precise molecular weight: 365.13 LC-MS (m/z): 366.08[M+H]+
1HNMR (400 MHz, DMSO-d6) δ (ppm): 8.25 (d, J=5.2 Hz, 1H), 8.12 (d, J=4.8 Hz, 1H), 7.92 (d, J=8.4 Hz, 1H), 7.60 (s, 1H), 7.45 (d, J=6.8 Hz, 2H), 4.48 (s, 1H), 3.20 (d, J=8.8 Hz, 1H), 2.86 (d, J=10.4 Hz, 1H), 2.37 (s, 3H), 2.23 (s, 2H), 2.01-1.98 (m, 3H), 1.82 (s, 1H), 1.72-1.63 (m, 1H), 1.54 (s, 1H).
Molecular formula: C19H19N5OS Precise molecular weight: 365.13 LC-MS (m/z): 366.14[M+H]+
1-Bromo-2-methoxy-4-methylbenzene (20.96 g, 104.22 mmol, 1.05 eq.) was dissolved in anhydrous tetrahydrofuran (150 mL), followed by dropwise addition of n-butyllithium (48 mL, 119.11 mmol, 1.2 eq.) at −78° C. After the completion of dropwise addition, the reaction solution was added dropwise to a solution of 3,4-pyridine dicarboxylic anhydride (14.8 g, 99.26 mmol, 1.0 eq.) in tetrahydrofuran (150 mL) at −78° C. After the completion of dropwise addition, the mixture was reacted at room temperature for 1 h. After the reaction was completed as detected by TLC, the reaction solution was poured into a mixed solution of ice water (200 mL) and 2 mol/L hydrochloric acid (200 mL). The mixture was concentrated under reduced pressure and extracted with dichloromethane (200 mL×3). The organic phases were combined, dried and concentrated to give the crude product (13.7 g), which was directly used in the next step.
The mixture obtained in the previous step was dissolved in ethanol (120 mL), followed by addition of 85% hydrazine hydrate (3.97 g, 67.4 mmol, 1.5 eq.). The mixture was heated to 80° C. and reacted for 2 h. After the reaction was completed as detected by LC-MS, the reaction solution was cooled to room temperature, and solids precipitated. The mixture was filtered, and the filtrate was concentrated and purified by silica gel column chromatography (dichloromethane:methanol=1200:1 to 200:1) to give the product (2.35 g, two-step yield: 8.8%).
Acetonitrile (20 mL) and phosphorus oxychloride (2.29 g, 14.96 mmol, 2.0 eq.) were added to 4-(2-methoxy-4-methylphenyl)pyrido[3,4-d]pyridazin-1-ol (2 g, 7.48 mmol, 1.0 eq.), and the mixture was reacted at 100° C. for 16 h. After there remained some of the materials as detected by TLC, the reaction solution was supplemented with phosphorus oxychloride (2.29 g, 14.96 mmol, 2.0 eq.) and reacted at 100° C. for 1 h. After the reaction was almost completed as detected by TLC, the reaction solution was concentrated, and the concentrated solution was poured into ice water (10 mL), followed by addition of dichloromethane (20 mL). The pH was adjusted to low alkalinity with sodium bicarbonate, followed by extraction with dichloromethane (20 mL×3). The organic phases were combined, dried, concentrated and purified by silica gel column chromatography (petroleum ether:ethyl acetate=6:1) to give the product (955 mg, yield: 44.6%).
1-Chloro-4-(2-methoxy-4-methylphenyl)pyrido[3,4-d]pyridazine (955 mg, 3.34 mmol, 1.0 eq.) was dissolved in DMAc (10 mL), followed by addition of (R)-1-tert-butoxycarbonyl-3-aminopiperidine (1.34 g, 6.68 mmol, 2.0 eq.). The mixture was reacted at 120° C. for 20 h. After the reaction was completed as detected by TLC, the reaction solution was poured into water (20 mL), followed by extraction with ethyl acetate (20 mL×2). The organic phases were combined, dried and concentrated, and the crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=5:1 to 1:1) to give the product (980 mg, yield: 65.3%).
Tert-butyl (R)-3-((4-(2-methoxy-4-methylphenyl)pyrido[3,4-d]pyridazin-1-yl)amino)piperidine-1-carboxylate (980 mg, 2.18 mmol, 1.0 eq.) was dissolved in dichloromethane (10 mL), followed by dropwise addition of boron tribromide (1.64 g, 6.54 mmol, 3.0 eq.) at 0° C. The mixture was reacted at room temperature for 2 h. After there remained some of the materials as detected by TLC, the reaction solution was supplemented with boron tribromide (1.64 g, 6.54 mmol, 3.0 eq.). The resulting mixture was reacted at room temperature for 2 h. After the reaction was almost completed as detected by TLC, methanol (5 mL) and water (10 mL) were added dropwise, slowly and successively to the reaction solution under an ice bath, followed by back-extraction with dichloromethane (20 mL). The pH of the aqueous phase was adjusted to alkalinity with a saturated aqueous sodium bicarbonate solution, followed by extraction with dichloromethane (10 mL). The aqueous phase was concentrated, dissolved in a mixed solution of dichloromethane/methanol, dried over anhydrous magnesium sulfate and concentrated, and the crude product was directly used in the next step.
(R)-5-methyl-2-(1-((piperidin-3-yl)amino)pyrido[3,4-d]pyridazin-4-yl)phenol (474 mg, 1.41 mmol, 1.0 eq.) was dissolved in methanol (5 mL), followed by addition of an aqueous formaldehyde solution (37%) (112 mg, 1.41 mmol, 1.0 eq.) and sodium cyanoborohydride (97 mg, 1.55 mmol, 1.1 eq.). The mixture was reacted at room temperature for 5 min. After the reaction was completed as detected by TLC, the reaction solution was concentrated, and the crude product was purified by preparative thin-layer chromatography (dichloromethane:methanol=7:1) to give the product (110 mg, two-step yield: 22.3%).
1H-NMR (300 MHz, DMSO-d6) δ (ppm): 9.71 (brs, 1H), 8.94-8.92 (d, 1H), 8.87 (s, 1H), 8.30-8.27 (d, 1H), 7.42-7.40 (d, 1H), 7.27-7.25 (d, 1H), 6.83-6.80 (d, 2H), 4.47 (s, 1H), 3.27-3.18 (m, 2H), 2.90-2.88 (m, 1H), 2.39-2.20 (m, 7H), 2.04-1.83 (m, 2H), 1.73-1.52 (m, 2H).
Molecular formula: C20H23N5O Precise molecular weight: 349.19 LC-MS (m/z): 350.10 [M+H]+
Methyl 5-chloro-2-iodobenzoate (10.3 g, 34.67 mmol, 1.0 eq.) was dissolved in anhydrous tetrahydrofuran (100 mL), followed by dropwise addition of a 1.3 mol/L solution of an isopropylmagnesium chloride lithium chloride complex in tetrahydrofuran (32 mL, 41.6 mmol, 1.2 eq.) at −28° C. After the completion of dropwise addition, the mixture was reacted at this temperature for 30 min. At −78° C., the reaction solution was added dropwise to a solution of 2-methoxy-4-methylbenzoyl chloride (9.6 g, 52 mmol, 1.5 eq.) in tetrahydrofuran (100 mL). After the completion of dropwise addition, the mixture was reacted at room temperature for 1 h. After the reaction was completed as detected by TLC, the reaction solution was concentrated and poured into ice water (100 mL), and potassium carbonate was added, followed by extraction with dichloromethane (200 mL×3). The organic phases were combined, dried and concentrated to give the product (10.3 g, yield: 93.2%).
4-(2-Methoxy-4-methylbenzoyl)nicotinic acid (10.3 g, 32.3 mmol, 1.0 eq.) was dissolved in ethanol (100 mL), followed by addition of 85% hydrazine hydrate (2.86 g, 48.45 mmol, 1.5 eq.). The mixture was heated to 90° C. and reacted for 3 h. After the reaction was completed as detected by TLC, the reaction solution was cooled to room temperature, and solids precipitated. The mixture was filtered, and the filter cake was the product (1.5 g, yield: 15.4%).
Acetonitrile (10 mL) and phosphorus oxychloride (1.53 g, 9.98 mmol, 2.0 eq.) were added to 7-chloro-4-(2-methoxy-4-methylphenyl)phthalazin-1(2H)-one (1.5 g, 4.99 mmol, 1.0 eq.), and the mixture was reacted at 90° C. for 3 h. After the reaction was completed as detected by TLC, the reaction solution was concentrated, the concentrated solution was poured into ice water (10 mL), and sodium bicarbonate was added to adjust the pH to low alkalinity, followed by extraction with dichloromethane (10 mL×3). The organic phases were combined, dried and concentrated to give the product (1.31 g, yield: 82.3%).
4,6-Dichloro-1-(2-methoxy-4-methylphenyl)phthalazine (1.31 g, 4.1 mmol, 1.0 eq.) was dissolved in DMAc (10 mL), (R)-1-tert-butyloxycarbonyl-3-aminopiperidine (1.64 g, 8.2 mmol, 2.0 eq.) was added, and the reaction solution was reacted at 120° C. for 24 h. After the reaction was completed as detected by TLC, the reaction solution was poured into water (20 mL), followed by extraction with ethyl acetate (20 mL×2). The organic phases were combined, dried, and concentrated, and the crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=10:1 to 1:1) to give the product (870 mg, yield: 40.9%).
Tert-butyl (R)-3-((7-(2-methoxy-4-methylphenyl)phthalazin-1-yl)amino)piperidine-1-carboxylate (300 mg, 0.621 mmol, 1.0 eq.) was dissolved in dichloromethane (5 mL), boron tribromide (467 mg, 1.863 mmol, 3.0 eq.) was added dropwise at 0° C., and the mixture was reacted at room temperature for 30 min. After the reaction was completed as detected by TLC, methanol (5 mL) and water (5 mL) were added dropwise and slowly to the reaction solution under an ice bath, followed by back-extraction with dichloromethane (20 mL×1, 10 mL×3). Sodium bicarbonate was added to the aqueous phase to adjust the pH to alkalinity, followed by extraction with dichloromethane:methanol=10:1 (10 mL×3), and the organic phases were combined, dried over anhydrous magnesium sulfate, and concentrated to give the product (78 mg, yield: 34%).
(R)-2-(6-chloro-4-((piperidin-3-yl)amino)phthalazin-1-yl)-5-methylphenol (78 mg, 0.211 mmol, 1.0 eq.) was dissolved in methanol (1 mL), followed by addition of an aqueous formaldehyde solution (37%) (17 mg, 0.211 mmol, 1.0 eq.) and sodium cyanoborohydride (15 mg, 0.232 mmol, 1.1 eq.), and the resulting mixture was reacted at room temperature for 5 min. After the reaction was completed as detected by TLC, the reaction solution was concentrated, and the crude product was purified by preparative thin-layer chromatography (dichloromethane:methanol=7:1) to give the product (27 mg, yield: 33.3%).
1H-NMR (300 MHz, DMSO-d6) δ (ppm): 9.61 (brs, 1H), 8.62 (s, 1H), 7.85-7.82 (dd, 1H), 7.53-7.50 (d, 2H), 7.14-7.17 (d, 1H), 6.82-6.76 (m, 2H), 4.65 (s, 1H), 3.60-3.56 (m, 3H), 2.73 (s, 4H), 2.32 (s, 3H), 2.03-2.01 (m, 2H), 1.83-1.80 (m, 2H).
Molecular formula: C21H23C1N4O Precise molecular weight: 382.16 LC-MS (m/z): 383.09/385.03 [M+H]+.
1-Bromo-2-methoxy-4-methylbenzene (10.0 g, 49.73 mmol, 1.0 eq.) was added to tetrahydrofuran (30.0 mL), and the mixture was cooled to −65° C. in nitrogen atmosphere, followed by dropwise and slow addition of a 2.5 mol/L solution of n-butyllithium in n-hexane (19.9 mL, 49.73 mmol, 1.0 eq.). The reaction solution was reacted for 1 h, and added slowly and dropwise to a solution of 4,5,6,7-tetrahydroisobenzofuran-1,3-dione (7.56 g, 49.73 mmol, 1.0 eq.) in tetrahydrofuran (30.0 mL) at −65° C., and the solution was warmed to room temperature and reacted for 12 h. After the reaction was completed as monitored by TLC, a 2 mol/L aqueous HCl solution was added to the system with stirring for 0.5 h, followed by extraction with ethyl acetate (200.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and the crude product was slurried with petroleum ether (100.0 mL) for 2 h, and filtered under vacuum. The filter cake was dried to give the product (5.88 g, yield: 43.1%).
2-(2-Methoxy-4-methylbenzoyl)cyclohex-1-ene-1-carboxylic acid (5.88 g, 21.43 mmol, 1.0 eq.) was added to N,N-dimethylformamide (20.0 mL), followed by addition of potassium carbonate (3.85 g, 27.85 mmol, 1.3 eq.) and iodomethane (3.65 g, 25.71 mmol, 1.2 eq.), and the resulting mixture was reacted at room temperature for 2 h. After the reaction was completed as monitored by TLC, ethyl acetate (200.0 mL) was added, followed by washing with water (100.0 mL×4). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to give the product (5.24 g, yield: 84.7%).
Methyl 2-(2-methoxy-4-methylbenzoyl)cyclohex-1-ene-1-carboxylate (5.24 g, 18.17 mmol, 1.0 eq.) was added to ethanol (80.0 mL), followed by addition of hydrazine hydrate (9.0 g, 181.7 mmol, 10.0 eq.), and the mixture was reacted at 80° C. for 12 h. After the reaction was completed as monitored by TLC, the system was concentrated under reduced pressure, and the crude product was separated by silica gel column chromatography (specification of silica gel: 100-200 mesh, petroleum ether:ethyl acetate=10:1 to 2:1) to give the product (820.0 mg, yield: 16.7%).
4-(2-Methoxy-4-methylphenyl)-5,6,7,8-tetrahydrophthalazin-1-ol (820.0 mg, 3.03 mmol, 1.0 eq.) was added to acetonitrile (10.0 mL), followed by addition of phosphorus oxychloride (2.78 g, 18.19 mmol, 6.0 eq.), and the mixture was stirred at 88° C. for 6 h. After the reaction was completed as monitored by TLC, the reaction solution was cooled to room temperature, and a saturated aqueous sodium bicarbonate solution was added to quench the reaction, followed by addition of ethyl acetate (100.0 mL) for extraction. The organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (specification of silica gel: 100-200 mesh, petroleum ether:ethyl acetate=2:1) to give the product (600.0 mg, yield: 68.5%).
1-Chloro-4-(2-methoxy-4-methylphenyl)-5,6,7,8-tetrahydrophthalazine (600.0 mg, 4.13 mmol, 1.0 eq.) was added to dichloromethane (10.0 mL), the mixture was cooled to −65° C., followed by dropwise and slow addition of boron tribromide (1.56 g, 6.23 mmol, 3.0 eq.), and the reaction solution was warmed to room temperature and stirred for 3 h. After the reaction was completed as monitored by TLC, methanol was added to the system to quench the reaction, followed by concentration under reduced pressure, and a saturated aqueous sodium bicarbonate solution was added to adjust the pH to 7-8, followed by extraction with dichloromethane (100.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to give the product (400.0 mg, yield: 70.4%).
2-(4-Chloro-5,6,7,8-tetrahydrophthalazin-1-yl)-5-methylphenol (370.0 mg, 1.34 mmol, 1.0 eq.), tert-butyl (R)-3-aminopiperidine-1-carboxylate (539.4 mg, 2.68 mmol, 2.0 eq.), sodium tert-butoxide (386.3 mg, 4.02 mmol, 3.0 eq.), Pd2(dba)3 (368.1 mg, 0.40 mmol, 0.3 eq.) and 1,1′-binaphthyl-2,2′-bisdiphenylphosphine (375.1 mg, 0.60 mmol, 0.45 eq.) were added to 1,4-dioxane (15.0 mL), followed by purge with nitrogen for 3 min, and the mixture was reacted at 90° C. for 12 h. After the reaction was completed as monitored by TLC, the system was cooled to room temperature and filtered, and the filtrate was concentrated under reduced pressure to give a crude product (600.0 mg, used directly in the next step).
Crude tert-butyl (R)-3-((4-(2-hydroxy-4-methylphenyl)-5,6,7,8-tetrahydrophthalazin-1-yl)amino)piperidine-1-carboxylate (600.0 mg, 1.34 mmol, 1.0 eq.) was added to a mixture of dichloromethane (6.0 mL) and trifluoroacetic acid (6 mL), and the mixture was reacted at room temperature for 1 h. After the reaction was completed as monitored by TLC, water (50.0 mL) was added for back-extraction, and the pH of the aqueous phase was adjusted to 8-9, followed by extraction with dichloromethane (50.0 mL×2). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to give the product (130.0 mg, two-step yield: 28.6%).
(R)-5-methyl-2-(4-(piperidin-3-ylamino)-5,6,7,8-tetrahydrophthalazin-1-yl)phenol (130.0 mg, 0.38 mmol, 1.0 eq.) was added to methanol (3.0 mL), and an aqueous formaldehyde solution (37%) (34.2 mg, 0.42 mmol, 1.1 eq.) was added. The mixture was stirred at room temperature for 1 h, followed by addition of sodium cyanoborohydride (31.0 mg, 0.49 mmol, 1.3 eq.), and the resulting mixture was reacted at room temperature for 2 h. After the reaction was completed as monitored by TLC, the system was concentrated under reduced pressure, and a saturated aqueous sodium bicarbonate solution (100.0 mL) was added, with stirring for 0.5 h, followed by extraction with dichloromethane (100.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and the crude product was purified by preparative thin-layer chromatography (dichloromethane:methanol=8:1) to give the product (30.0 mg, yield: 22.4%).
1HNMR (300 MHz, DMSO-d6) δ (ppm): 9.63 (s, 1H), 6.98-7.00 (m, 1H), 6.67-6.72 (m, 2H), 5.71 (s, 1H), 4.37 (s, 1H), 2.89-3.20 (m, 3H), 2.37-2.45 (m, 6H), 2.27 (s, 3H), 1.59-1.85 (m, 7H), 1.24 (s, 3H).
Molecular formula: C21H28N4O Molecular weight: 352.48 LC-MS (Pos, m/z)=353.26 [M+H]+.
1-Bromo-2-methoxy-4-methylbenzene (7.27 g, 36.2 mmol, 1.0 eq.) was added to dry tetrahydrofuran (30.0 mL), and the mixture was cooled to −65° C. in nitrogen atmosphere, followed by dropwise and slow addition of a 2.5 mol/L solution of n-butyllithium in n-hexane (14.5 mL, 36.2 mmol, 1.0 eq.). After the reaction was carried out for 1 h, the reaction solution was added dropwise and slowly to a solution of 5,6-dihydro-1H-cyclopenta[c]furan-1,3(4H)-dione (5.0 g, 36.2 mmol, 1.0 eq.) in tetrahydrofuran (20.0 mL) at −65° C., and the solution was warmed to room temperature for 12 h. After the reaction was completed as monitored by TLC, a 2 mol/L aqueous HCl solution was added to the system with stirring for 0.5 h, followed by extraction with ethyl acetate (200.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to give the crude product (7.0 g) which was used directly in the next step.
2-(2-Methoxy-4-methylbenzoyl)cyclopent-1-ene-1-carboxylic acid (7.0 g, 26.9 mmol, 1.0 eq.), potassium carbonate (5.57 g, 40.35 mmol, 1.5 eq.) and iodomethane (4.97 g, 35.0 mmol, 1.3 eq.) were added to N,N-dimethylformamide (20.0 mL), and the mixture was reacted at room temperature for 12 h. After the reaction was completed as monitored by TLC, ethyl acetate (200.0 mL) was added to the system, followed by washing with water (100.0 mL×4). The organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (specification of silica gel: 100-200 mesh, petroleum ether:ethyl acetate=20:1 to 2:1) to give the product (5.0 g, two-step yield: 68.4%).
Methyl 2-(2-methoxy-4-methylbenzoyl)cyclopent-1-ene-1-carboxylate (4.75 g, 17.31 mmol, 1.0 eq.) was added to ethanol (50.0 mL), followed by addition of hydrazine hydrate (5.2 g, 103.89 mmol, 6.0 eq.), and the mixture was reacted at room temperature for 12 h and then reacted at 80° C. for 8 h. After the reaction was completed as monitored by TLC, the system was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (specification of silica gel: 100-200 mesh, dichloromethane:methanol=20:1 to 10:1) to give the product (3.2 g, yield: 72.2%).
4-(2-Methoxy-4-methylphenyl)-6,7-dihydro-5H-cyclopenta[d]pyridazin-1-ol (3.2 g, 12.48 mmol, 1.0 eq.) and phosphorus oxychloride (11.4 g, 74.88 mmol, 6.0 eq.) were added to acetonitrile (20.0 mL), and the mixture was stirred at 80° C. for 2 h. After the reaction was completed as monitored by TLC, the reaction solution was cooled to room temperature, a saturated aqueous sodium bicarbonate solution was added to carefully quench the reaction, and ethyl acetate (100.0 mL) was added to the system for extraction. The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to give the product (2.8 g, yield: 81.8%).
1-Chloro-4-(2-methoxy-4-methylphenyl)-6,7-dihydro-5H-cyclopenta[d]pyridazine (2.2 g, 8.0 mmol, 1.0 eq.) was added to 1,4-dioxane (20.0 mL), then tert-butyl (R)-3-aminopiperidine-1-carboxylate (3.2 g, 16.0 mmol, 2.0 eq.), cesium carbonate (7.8 g, 24.0 mmol, 3.0 eq.), Pd2(dba)3 (732.5 mg, 0.80 mmol, 0.1 eq.) and 1,1′-binaphthyl-2,2′-bisdiphenylphosphine (996.2 mg, 1.6 mmol, 0.2 eq.) were added successively, and the resulting mixture was reacted at 90° C. for 12 h in nitrogen atmosphere. After the reaction was completed as monitored by TLC, the system was cooled to room temperature and filtered, the filtrate was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (specification of silica gel: 100-200 mesh, petroleum ether:ethyl acetate=10:1 to 1:1) to give the product (2.8 g, yield: 80%).
Tert-butyl (R)-3-((4-(2-methoxy-4-methylphenyl)-6,7-dihydro-5H-cyclopenta[d]pyridazin-1-yl)amino)piperidine-1-carboxylate (2.8 g, 6.38 mmol, 1.0 eq.) and trifluoroacetic acid (10.0 mL) were added to dichloromethane (10.0 mL), and the mixture was reacted at room temperature for 2 h. After the reaction was completed as monitored by TLC, a saturated aqueous sodium carbonate solution was added to the system to adjust the pH to 8-9, followed by extraction with dichloromethane (100.0 mL×2). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to give the product (2.0 g, yield: 93.0%).
(R)-4-(2-methoxy-4-methylphenyl)-N-(piperidin-3-yl)-6,7-dihydro-5H-cyclopenta[d]pyridazin-1-amine (500.0 mg, 1.47 mmol, 1.0 eq.) and an aqueous formaldehyde solution (37%) (143.8 mg, 1.77 mmol, 1.2 eq.) were added to methanol (5.0 mL), and after stirring at room temperature for 1 h, sodium cyanoborohydride (129.3 mg, 2.05 mmol, 1.4 eq.) was added, and the resulting mixture was reacted at room temperature for 1 h. After the reaction was completed as monitored by TLC, the system was concentrated under reduced pressure, and a saturated aqueous sodium bicarbonate solution (100.0 mL) was added, with stirring for 0.5 h, followed by extraction with dichloromethane (100.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (specification of silica gel: 100-200 mesh, dichloromethane:methanol=50:1 to 10:1) to give the product (370.0 mg, yield: 71.4%).
(R)-4-(2-methoxy-4-methylphenyl)-N-(1-methylpiperidin-3-yl)-6,7-dihydro-5H-cyclopenta[d]pyridazin-1-amine (370.0 mg, 1.04 mmol, 1.0 eq.) was added to dichloromethane (50.0 mL), the mixture was cooled to −50° C., followed by dropwise and slow addition of boron tribromide (788.9 mg, 3.14 mmol, 3.0 eq.), and the reaction solution was warmed to room temperature and stirred for 12 h. After the reaction was completed as monitored by TLC, methanol was added to the system to quench the reaction, followed by concentration under reduced pressure, and a saturated aqueous sodium bicarbonate solution was added to adjust the pH to 7-8, followed by extraction with dichloromethane (100.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and the crude product was purified by preparative thin-layer chromatography (dichloromethane:methanol=8:1) to give the product (165.0 mg, yield: 46.8%).
1HNMR (300 MHz, DMSO-d6) δ (ppm): 7.48-7.51 (d, J=9 Hz, 1H), 6.91 (s, 1H), 6.70-6.73 (m, 1H), 4.70 (s, 1H), 3.26-3.31 (m, 2H), 2.86-2.98 (m, 4H), 2.71-2.74 (m, 1H), 2.48 (s, 3H), 2.40-2.42 (m, 1H), 2.35 (s, 3H), 2.17-2.26 (m, 2H), 1.69-2.06 (m, 4H).
Molecular formula: C20H26N4O Molecular weight: 338.46 LC-MS (Pos, m/z)=339.17 [M+H]+.
1,4-Dichloro-5,6,7,8-tetrahydrophthalazine (613.3 mg, 3.02 mmol, 1.0 eq.), (4-cyclopropyl-2-methoxyphenyl)boronic acid (580.0 mg, 3.02 mmol, 1.0 eq.), sodium bicarbonate (507.5 mg, 6.04 mmol, 2.0 eq.) and Pd(pph3)4 (348.9 mg, 0.30 mmol, 0.1 eq.) were added to 1,4-dioxane (10.0 mL) and water (5.0 mL), and the mixture was reacted at 110° C. for 2 h in nitrogen atmosphere. After the reaction was completed as monitored by TLC, the system was cooled to room temperature, followed by addition of ethyl acetate (100.0 mL) and washing with water (50.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (specification of silica gel: 100-200 mesh, petroleum ether:ethyl acetate=10:1 to 4:1) to give the product (500.0 mg, yield: 52.6%).
1-Chloro-4-(4-cyclopropyl-2-methoxyphenyl)-5,6,7,8-tetrahydrophthalazine (500.0 mg, 1.58 mmol, 1.0 eq.), tert-butyl (R)-3-aminopiperidine-1-carboxylate (636.1 mg, 3.17 mmol, 2.0 eq.), cesium carbonate (1.54 g, 4.74 mmol, 3.0 eq.), Pd2(dba)3 (144.6 mg, 0.15 mmol, 0.1 eq.) and BINAP (196.7 mg, 0.31 mmol, 0.2 eq.) were added to 1,4-dioxane (15.0 mL), and the mixture was reacted at 90° C. for 12 h in nitrogen atmosphere. After the reaction was completed as monitored by TLC, the reaction solution was cooled, followed by addition of ethyl acetate (200.0 mL) and washing with water (100.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (specification of silica gel: 100-200 mesh, dichloromethane:methanol=100:1 to 80:1) to give the product (680.0 mg, yield: 89.9%).
Tert-butyl (R)-3-((4-(4-cyclopropyl-2-methoxyphenyl)-5,6,7,8-tetrahydrophthalazin-1-yl)amino)piperidine-1-carboxylate (680.0 mg, 1.42 mmol, 1.0 eq.) and trifluoroacetic acid (5.0 mL) were added to dichloromethane (10.0 mL), and the mixture was stirred at room temperature for 2 h. After the reaction was completed as monitored by TLC, a saturated aqueous sodium bicarbonate solution was added to the system to adjust the pH to 7-8, followed by extraction with dichloromethane (100.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to give the product (530.0 mg, yield: 98.6%).
(R)-4-(4-cyclopropyl-2-methoxyphenyl)-N-(piperidin-3-yl)-5,6,7,8-tetrahydrophthalazin-1-amine (530.0 mg, 1.40 mmol, 1.0 eq.) and an aqueous formaldehyde solution (37%) (136.3 mg, 1.68 mmol, 1.2 eq.) were added to methanol (10.0 mL), the mixture was stirred at room temperature for 0.5 h, followed by addition of sodium cyanoborohydride (123.1 mg, 1.96 mmol, 1.4 eq.), and the reaction solution was then reacted at room temperature for 2 h. After the reaction was completed as monitored by TLC, the system was concentrated under reduced pressure, followed by addition of a saturated aqueous sodium bicarbonate solution (100.0 mL). The system was stirred for 30 min and extracted with dichloromethane (50.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (specification of silica gel: 100-200 mesh, dichloromethane:methanol=50:1 to 10:1) to give the product (412.1 mg, yield: 75.0%).
(R)-4-(4-cyclopropyl-2-methoxyphenyl)-N-(1-methylpiperidin-3-yl)-5,6,7,8-tetrahydrophthalazin-1-amine (400.0 mg, 1.01 mmol, 1.0 eq.) was added to dichloromethane (4.0 mL), boron tribromide (765.8 mg, 3.03 mmol, 3.0 eq.) was added dropwise under an ice bath, and the reaction solution was warmed to room temperature and stirred for 4 h. After the reaction was completed as monitored by TLC, methanol (5.0 mL) was added under an ice bath to quench the reaction, followed by concentration under reduced pressure, addition of dichloromethane (100.0 mL) and back extraction with water (50.0 mL). A saturated aqueous sodium bicarbonate solution was added to the aqueous phase to adjust the pH to 7-8, followed by extraction with dichloromethane (100.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and the crude product was purified by preparative thin-layer chromatography to give the product (170.0 mg, yield: 44.4%).
1HNMR (300 MHz, DMSO-d6) δ (ppm): 9.66 (s, 1H), 7.08-6.98 (m, 1H), 6.87-6.56 (m, 2H), 5.53-5.50 (m, 1H), 4.26 (s, 1H), 2.95-2.93 (m, 1H), 2.65-2.62 (m, 1H), 2.35-2.33 (m, 4H), 2.23 (s, 3H), 2.04-2.01 (m, 2H), 1.87-1.69 (m, 6H), 1.58-1.47 (m, 4H), 0.96-0.93 (m, 2H), 0.86-0.83 (m, 1H), 0.66-0.63 (m, 1H).
Molecular formula: C23H30N4O Precise molecular weight: 378.24 LC-MS (m/z)=379.17 [M+H]+.
1,4-Dichloro-5,6,7,8-tetrahydro-5,8-ethanophthalazine (4.50 g, 19.6 mmol, 1.0 eq.), tert-butyl (R)-3-aminopiperidine-1-carboxylate (4.32 g, 21.6 mmol, 1.1 eq.), Pd2(dba)3 (1.79 g, 1.96 mmol, 0.1 eq.), BINAP (2.44 g, 3.92 mmol, 0.2 eq.) and Cs2CO3 (12.8 g, 39.2 mmol, 2.0 eq.) were added successively to 1,4-dioxane (100 mL), and the mixture was heated to 90° C. and reacted for 20 h in nitrogen atmosphere. The system was cooled to room temperature, and water was added to quench the reaction, followed by extraction with EA (100 mL×2). The organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (PE:EA=10:1 to 1:1) to give the product (5.90 g, yield: 76.4%).
Tert-butyl (R)-3-((4-chloro-5,6,7,8-tetrahydro-5,8-ethanophthalazin-1-yl)amino)piperidine-1-carboxylate (5.90 g, 15.0 mmol, 1.0 eq.) was dissolved in DCM (50 mL), then a solution of hydrogen chloride in 1,4-dioxane (4 mol/L, 37.5 mL, 150 mmol, 10.0 eq.) was added dropwise, and the reaction solution was stirred at room temperature for 4 h. Then water (80 mL) was added, followed by liquid separation. The organic phase was removed, and a saturated aqueous NaHCO3 solution was added to the aqueous phase to adjust the pH to 8, followed by extraction with DCM (50 mL×5). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to give the product (3.60 g, yield: 81.9%).
(R)-4-chloro-N-(piperidin-3-yl)-5,6,7,8-tetrahydro-5,8-ethanophthalazin-1-amine (3.60 g, 12.3 mmol, 1.0 eq.) was dissolved in MeOH (40 mL), an aqueous formaldehyde solution (37%, 1.20 g, 14.8 mmol, 1.2 eq.) was added, and the reaction solution was stirred at room temperature for 30 min. NaBH3CN (930 mg, 14.8 mmol, 1.2 eq.) was then added, and the reaction solution was reacted at room temperature for 1 h. MeOH was removed by concentration, the system was then dissolved in DCM, and a saturated aqueous NaHCO3 solution was added for washing. The aqueous phase was extracted with DCM (50 mL×2), the organic phases were combined, dried over anhydrous sodium sulfate, and filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (DCM:MeOH=50:1 to 20:1) to give the product (3.20 g, yield: 84.8%).
(2-Methoxy-4-methylphenyl)boronic acid (200 mg, 1.20 mmol, 1.0 eq.), (R)-4-chloro-N-(1-methylpiperidin-3-yl)-5,6,7,8-tetrahydro-5,8-ethanophthalazin-1-amine (370 mg, 1.20 mmol, 1.0 eq.), Pd(dppf)Cl2 (43.9 mg, 0.060 mmol, 0.05 eq.) and NaHCO3 (202 mg, 2.40 mmol, 2.0 eq.) were added successively to 1,4-dioxane (6 mL), then H2O (3 mL) was added, and the reaction solution was heated to 110° C. for 2 h in nitrogen atmosphere. The system was cooled to room temperature, and water (30 mL) was added to quench the reaction, followed by extraction with DCM (30 mL×2). The organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (DCM:MeOH=50:1 to 10:1) to give the product (320 mg, yield: 67.7%).
(R)-4-(2-methoxy-4-methylphenyl)-N-(1-methylpiperidin-3-yl)-5,6,7,8-tetrahydro-5,8-ethanophthalazin-1-amine (320 mg, 0.815 mmol, 1.0 eq.) was dissolved in DCM (10 mL), the reaction solution was cooled to −10° C., followed by dropwise addition of BBr3 (1.02 g, 4.07 mmol, 5.0 eq.), and the solution was naturally warmed to room temperature and reacted for 16 h. MeOH (10 mL) was added to quench the reaction, a saturated aqueous NaHCO3 solution was added to adjust the pH to 8, and DCM (30 mL×3) was added for extraction. The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (DCM:MeOH=50:1 to 10:1) to give the product (160 mg, yield: 51.9%).
1HNMR (400 MHz, DMSO-d6) δ (ppm): 9.90 (s, 1H), 7.10 (d, J=7.6 Hz, 1H), 6.76 (s, 1H), 6.71 (d, J=7.7 Hz, 1H), 6.41 (s, 1H), 4.43 (s, 1H), 3.45 (s, 1H), 3.06 (s, 1H), 2.83 (s, 1H), 2.59 (s, 4H), 2.28 (s, 3H), 1.92 (d, J=3.3 Hz, 2H), 1.74-1.66 (m, 7H), 1.74-1.24 (s, 5H).
Molecular formula: C23H30N4O Molecular weight: 378.24 LC-MS (m/z)=379.19 [M+H]+.
3-(Ethoxymethoxy)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzonitrile (300 mg, 0.990 mmol, 1.0 eq.), (R)-4-chloro-N-(1-methylpiperidin-3-yl)-5,6,7,8-tetrahydro-5,8-ethanophthalazin-1-amine (304 mg, 0.990 mmol, 1.0 eq.), Pd(dppf)Cl2 (36.2 mg, 0.0495 mmol, 0.05 eq.) and NaHCO3 (166 mg, 1.98 mmol, 2.0 eq.) were added successively to 1,4-dioxane (6 mL) and H2O (3 mL), and the mixture was heated to 110° C. for 2 h in nitrogen atmosphere. The system was cooled to room temperature, and water (30 mL) was added to quench the reaction, followed by extraction with DCM (30 mL×2). The organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (DCM:MeOH=50:1 to 10:1) to give the product (265 mg, yield: 59.8%).
(R)-3-(ethoxymethoxy)-4-(4-((1-methylpiperidin-3-yl)amino)-5,6,7,8-tetrahydro-5,8-ethanophthalazin-1-yl)benzonitrile (265 mg, 0.592 mmol, 1.0 eq.) was dissolved in DCM (4 mL), then a solution of hydrogen chloride in 1,4-dioxane (4 mol/L, 0.74 mL, 2.96 mmol, 5.0 eq.) was added dropwise, and the reaction solution was stirred at room temperature for 1 h. Water (20 mL) was added to quench the reaction, followed by washing with DCM (20 mL×2), and a saturated aqueous NaHCO3 solution was added to the aqueous phase to adjust the pH to 8, followed by extraction with DCM (20 mL×2). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure and slurried with EA to give the product (172 mg, yield: 74.6%).
1HNMR (400 MHz, DMSO-d6) δ (ppm): 7.40 (d, J=7.5 Hz, 1H), 7.29 (s, 1H), 7.21 (s, 1H), 6.14 (s, 1H), 4.20 (s, 1H), 3.57 (s, 1H), 2.99 (s, 1H), 2.72 (s, 1H), 2.65 (s, 1H), 2.18 (s, 3H), 1.88-1.57 (m, 9H), 1.35-1.18 (m, 5H).
Molecular formula: C23H27N5O Precise molecular weight: 389.22 LC-MS (m/z)=390.14 [M+H]+.
(2-(Ethoxymethoxy)-4-(trifluoromethyl)phenyl)boronic acid (1.00 g, 3.79 mmol, 1.0 eq.), 1,4-dichloro-5,6,7,8-tetrahydrophthalazine (769 mg, 3.79 mmol, 1.0 eq.), Pd(dppf)Cl2 (139 mg, 0.190 mmol, 0.05 eq.) and NaHCO3 (637 mg, 7.58 mmol, 2.0 eq.) were added successively to 1,4-dioxane (10 mL), then H2O (5 mL) was added, and the reaction solution was heated to 110° C. and reacted for 3 h in nitrogen atmosphere. The system was cooled to room temperature, and water (40 mL) was added to quench the reaction, followed by extraction with EA (30 mL×2). The organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (PE:EA=20:1 to 5:1) to give the product (890 mg, yield: 60.7%).
1-Chloro-4-(2-(ethoxymethoxy)-4-(trifluoromethyl)phenyl)-5,6,7,8-tetrahydrophthalazine (890 mg, 2.30 mmol, 1.0 eq.), tert-butyl (R)-3-aminopiperidine-1-carboxylate (691 mg, 2.45 mmol, 1.5 eq.), Pd2(dba)3 (211 mg, 0.230 mmol, 0.1 eq.), BINAP (286 mg, 0.460 mmol, 0.2 eq.) and Cs2CO3 (1.50 g, 4.60 mmol, 2.0 eq.) were added successively to 1,4-dioxane (20 mL), and the reaction solution was heated to 90° C. and reacted for 4 h in nitrogen atmosphere. The system was cooled to room temperature, and water (50 mL) was added to quench the reaction, followed by extraction with EA (40 mL×2). The organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (DCM:MeOH=100:1 to 50:1) to give the product (1.10 g, yield: 86.8%).
Tert-butyl (R)-3-((4-(2-(ethoxymethoxy)-4-(trifluoromethyl)phenyl)-5,6,7,8-tetrahydrophthalazin-1-yl)amino)piperidine-1-carboxylate (1.10 g, 2.00 mmol, 1.0 eq.) was dissolved in DCM (10 mL), then a solution of hydrogen chloride in 1,4-dioxane (4 mol/L, 5.0 mL, 20.0 mmol, 10.0 eq.) was added dropwise, and the reaction solution was stirred at room temperature for 1 h. Then water (30 mL) was added to quench the reaction, followed by washing with DCM (30 mL×2), and a saturated aqueous NaHCO3 solution was added to the aqueous phase to adjust the pH to 8, followed by extraction with DCM (30 mL×2). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to give the product (620 mg, yield: 79.1%).
(R)-2-(4-(piperidin-3-ylamino)-5,6,7,8-tetrahydrophthalazin-1-yl)-5-(trifluoromethyl)phenol (620 mg, 1.58 mmol, 1.0 eq.) was dissolved in MeOH (10 mL), then an aqueous formaldehyde solution (37%, 154 mg, 1.90 mmol, 1.2 eq.) was added, and the reaction solution was stirred at room temperature for 20 min. NaBH3CN (119 mg, 1.90 mmol, 1.2 eq.) was then added, and the reaction solution was reacted at room temperature for 1 h. After concentration, the system was then dissolved in DCM, and a saturated aqueous NaHCO3 solution was added for washing. The aqueous phase was extracted with DCM (20 mL×2), the organic phases were combined, dried over anhydrous sodium sulfate, and filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (DCM:MeOH=50:1 to 20:1) to give the product (410 mg, yield: 63.9%).
1HNMR (400 MHz, DMSO-d6) δ (ppm): 10.43 (s, 1H), 7.34 (d, J=7.5 Hz, 1H), 7.21 (d, J=7.5 Hz, 2H), 5.63 (d, J=7.8 Hz, 1H), 4.30-4.24 (m, 1H), 2.93 (d, J=8.4 Hz, 1H), 2.61 (s, 1H), 2.38-2.35 (m, 2H), 2.32-2.30 (m, 2H), 2.21 (s, 3H), 1.99 (d, J=8.8 Hz, 2H), 1.83 (d, J=8.8 Hz, 1H), 1.76-1.69 (m, 3H), 1.61-1.54 (m, 3H), 1.48-1.43 (m, 1H).
Molecular formula: C21H25F3N4O Precise molecular weight: 406.20 LC-MS (m/z)=407.12 [M+H]+.
(R)-3-(ethoxymethoxy)-4-(4-(piperidin-3-ylamino)-5,6,7,8-tetrahydrophthalazin-1-yl)benzonitrile (317.0 mg, 0.77 mmol, 1.0 eq.), triethylamine (116.8 mg, 1.15 mmol, 1.5 eq.) and methanesulfonic anhydride (176.1 mg, 1.01 mmol, 1.3 eq.) were added to dichloromethane (10.0 mL), and the reaction solution was reacted at room temperature for 12 h. After the reaction was completed as monitored by TLC, a saturated aqueous ammonium chloride solution (50.0 mL) was added, then dichloromethane (50.0 mL) was added for extraction, and the organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to give the product (343.0 mg, yield: 91.9%).
(R)-3-(ethoxymethoxy)-4-(4-((1-(methylsulfonyl)piperidin-3-yl)amino)-5,6,7,8-tetrahydrophthalazin-1-yl)benzonitrile (343.0 mg, 0.70 mmol, 1.0 eq.) and a solution of hydrogen chloride in 1,4-dioxane (4.0 mol/L, 3.0 mL) were added to dichloromethane (3.0 mL), and the reaction solution was reacted at room temperature for 2 h. After the reaction was completed as monitored by TLC, a saturated aqueous sodium bicarbonate solution was added to the system to adjust the pH to 7-8, followed by extraction with dichloromethane (50.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and the crude product was purified by preparative thin-layer chromatography to give the product (207.0 mg, yield: 69.2%).
1HNMR (400 MHz, DMSO-d6) δ (ppm): 10.50 (s, 1H), 7.33 (s, 2H), 7.24 (s, 1H), 5.84-5.82 (d, J=4 Hz, 1H), 4.26-4.24 (m, 1H), 3.88-3.84 (m, 1H), 3.52-3.47 (m, 1H), 2.88 (s, 3H), 2.75-2.70 (m, 1H), 2.58-2.53 (m, 1H), 2.40-2.37 (m, 2H), 2.31-2.28 (m, 2H), 1.97-1.95 (m, 1H), 1.88-1.86 (m, 1H), 1.77-1.75 (m, 2H), 1.61-1.60 (m, 4H).
Molecular formula: C21H25N5O3S Precise molecular weight: 427.17 LC-MS (m/z)=428.1 [M+H]+.
1,4-Dichloro-5,6,7,8-tetrahydrophthalazine (3.0 g, 14.77 mmol, 1.0 eq.), 3-methoxy-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzonitrile (3.0 g, 11.81 mmol, 0.8 eq.), sodium bicarbonate (2.48 g, 29.54 mmol, 2.0 eq.) and Pd(PPh3)4 (853.3 mg, 0.73 mmol, 0.05 eq.) were added to a mixed solution of 1,4-dioxane (20.0 mL) and water (10.0 mL), and the reaction solution was reacted at 110° C. for 12 h in nitrogen atmosphere. After the reaction was completed as monitored by TLC, water (100.0 mL) was added, and ethyl acetate (100.0 mL) was added for extraction. The organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=15:1 to 3:1) to give the product (2.2 g, yield: 49.7%).
4-(4-Chloro-5,6,7,8-tetrahydrophthalazin-1-yl)-3-methoxybenzonitrile (1.5 g, 5.00 mmol, 1.0 eq.), tert-butyl (R)-3-aminopiperidine-1-carboxylate (2.0 g, 10.00 mmol, 2.0 eq.), cesium carbonate (4.88 g, 15.00 mmol, 3.0 q.), Pd2(dba)3 (457.8 mg, 0.50 mmol, 0.1 eq.) and 1,1′-binaphthyl-2,2′-bisdiphenylphosphine (622.67 mg, 1.00 mmol, 0.2 eq.) were added to 1,4-dioxane (30.0 mL), and the reaction solution was reacted at 90° C. for 12 h in nitrogen atmosphere. After the reaction was completed as monitored by TLC, water (100.0 mL) was added, and ethyl acetate (100.0 mL) was added for extraction. The organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (specification of silica gel: 100-200 mesh, dichloromethane:methanol=100:1 to 80:1) to give the product (1.1 g, yield: 47.6%).
Tert-butyl (R)-3-((4-(4-cyano-2-methoxyphenyl)-5,6,7,8-tetrahydrophthalazin-1-yl)amino)piperidine-1-carboxylate (1.0 g, 2.15 mmol, 1.0 eq.) was added to dichloromethane (10.0 mL), then trifluoroacetic acid (5.0 mL) was added dropwise, and the reaction solution was reacted at room temperature for 2 h. After the reaction was completed as monitored by TLC, a saturated aqueous sodium bicarbonate solution was added to the system to adjust the pH to 7-8, followed by extraction with dichloromethane (100.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to give the product (750.0 mg, yield: 96.0%).
(R)-3-methoxy-4-(4-(piperidin-3-ylamino)-5,6,7,8-tetrahydrophthalazin-1-yl)benzonitrile (750.0 mg, 2.06 mmol, 1.0 eq.) and an aqueous formaldehyde solution (37%) (167.4 mg, 2.06 mmol, 1.0 eq.) were added to methanol (10.0 mL), the reaction solution was stirred at room temperature for 0.5 h, followed by addition of sodium cyanoborohydride (168.2 mg, 2.67 mmol, 1.3 eq.), and then the solution was reacted at room temperature for 2 h. After the reaction was completed as monitored by TLC, the system was concentrated under reduced pressure, and a saturated aqueous sodium bicarbonate solution (100.0 mL) was added, with stirring for 0.5 h, followed by extraction with dichloromethane (100.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (dichloromethane:methanol=40:1 to 30:1) to give the product (600.0 mg, yield: 77.2%).
(R)-3-methoxy-4-(4-((1-methylpiperidin-3-yl)amino)-5,6,7,8-tetrahydrophthalazin-1-yl)benzonitrile (247.0 mg, 0.65 mmol, 1.0 eq.) was added to dichloromethane (10.0 mL), then boron tribromide (491.7 mg, 1.96 mmol, 3.0 eq.) was added dropwise under an ice bath, and the reaction solution was heated to room temperature for 4 h. After the reaction was completed as monitored by TLC, methanol (10.0 mL) was added to the system under an ice bath to quench the reaction, followed by concentration under reduced pressure, addition of dichloromethane (50.0 mL), and back extraction with water (50.0 mL). A saturated aqueous sodium bicarbonate solution was added to the aqueous phase to adjust the pH to 7-8, followed by extraction with dichloromethane (100.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and the crude product was purified by preparative thin-layer chromatography to give the product (130.0 mg, yield: 55.0%).
1HNMR (300 MHz, DMSO-d6) δ (ppm): 10.59 (s, 1H), 7.33-7.30 (m, 3H), 5.94 (s, 1H), 4.46 (s, 1H), 2.99 (s, 1H), 2.55-2.51 (s, 3H), 2.42-2.29 (m, 6H), 1.88-1.69 (m, 5H), 1.61-1.60 (m, 4H).
Molecular formula: C21H25N5O Precise molecular weight: 363.21 LC-MS (m/z)=364.14 [M+H]+.
4-(4-Chloro-5,6,7,8-tetrahydrophthalazin-1-yl)-3-(ethoxymethoxy)benzonitrile (400.0 mg, 1.16 mmol, 1.0 eq.), (1R,2R)-2-aminocyclohexan-1-ol (267.9 mg, 2.32 mmol, 2.0 eq.), cesium carbonate (1.22 g, 3.48 mmol, 3.0 q.), Pd2(dba)3 (106.2 mg, 0.11 mmol, 0.1 eq.) and 1,1′-binaphthyl-2,2′-bisdiphenylphosphine (144.4 mg, 0.22 mmol, 0.2 eq.) were added to 1,4-dioxane (20.0 mL), and the reaction solution was reacted at 90° C. for 12 h in nitrogen atmosphere. After the reaction was completed as monitored by TLC, water (100.0 mL) was added, and ethyl acetate (100.0 mL) was added for extraction. The organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (specification of silica gel: 100-200 mesh, dichloromethane:methanol=80:1 to 50:1) to give the product (251.0 mg, yield: 51.2%).
3-(Ethoxymethoxy)-4-(4-(((1R,2R)-2-hydroxycyclohexyl)amino)-5,6,7,8-tetrahydrophthalazin-1-yl)benzonitrile (251.0 mg, 0.59 mmol, 1.0 eq.) was added to ethanol (3.0 mL), a hydrogen chloride-ethanol solution (2.0 mol/L, 8.0 mL) was added dropwise, and the reaction solution was reacted at room temperature for 2 h. After the reaction was completed as monitored by TLC, a saturated aqueous sodium bicarbonate solution was added to the system to adjust the pH to 7-8, followed by extraction with dichloromethane (100.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and the crude product was purified by preparative thin-layer chromatography (dichloromethane:methanol=10:1) to give the product (77.0 mg, yield: 35.5%).
1HNMR (400 MHz, DMSO-d6) δ (ppm): 10.55 (s, 1H), 7.33-7.32 (m, 2H), 7.25-7.24 (m, 1H), 5.62-5.60 (d, J=8 Hz, 1H), 4.81 (s, 1H), 3.93-3.87 (m, 1H), 3.52-3.47 (m, 1H), 2.42-2.29 (m, 5H), 2.11-2.08 (m, 1H), 1.99-1.92 (m, 1H), 1.77-1.59 (m, 8H).
Molecular formula: C21H24N4O2 Precise molecular weight: 364.19 LC-MS (m/z)=365.12 [M+H]+.
4,5,6,7-Tetrahydroisobenzofuran-1,3-dione (50 g, 329 mmol, 1.0 eq.) was dissolved in ethanol (500 mL), then hydrazine hydrate (20.64 g, 413 mmol, 1.25 eq.) was added, and the reaction solution was stirred at room temperature overnight. After the reaction was completed as detected by TLC, the reaction solution was filtered under vacuum, and the filter cake was dried to give the product (47 g, yield: 86.0%).
2,3,5,6,7,8-Hexahydrophthalazine-1,4-dione (25 g, 150 mmol, 1.0 eq.) obtained in the previous step was dissolved in acetonitrile (250 mL), then phosphorus oxychloride (138 g, 904 mmol, 6.0 eq.) was added, and the reaction solution was stirred at 80° C. for 4 h. After the reaction was completed as detected by TLC, the reaction solution was slowly poured into ice water, and a large number of solids precipitated. After stirring for 1 h, the solution was filtered under vacuum, and the filter cake was dried to give the product (24 g, yield: 78.8%).
1,4-Dichloro-5,6,7,8-tetrahydrophthalazine (15 g, 73.9 mmol, 1.0 eq.) obtained in the previous step was dissolved in toluene (300 mL), then tert-butyl (R)-3-aminopiperidine-1-carboxylate (29.6 g, 147.8 mmol, 2.0 eq.), BINAP (9.2 g, 14.78 mmol, 0.2 eq.), cesium carbonate (72.3 g, 221.7 mmol, 3.0 eq.) and Pd2(dba)3 (6.8 g, 7.39 mmol, 0.1 eq.) were added successively, and the reaction solution was stirred at 90° C. overnight in nitrogen atmosphere. After the reaction was completed as detected by TLC, the reaction solution was filtered through celite under vacuum, the filtrate was concentrated, and the crude product was purified by silica gel column chromatography (dichloromethane:methanol=50:1 to 30:1) to give the product (13 g, yield: 43.9%).
Tert-butyl (R)-3-((4-chloro-5,6,7,8-tetrahydrophthalazin-1-yl)amino)piperidine-1-carboxylate (13 g, 35.5 mmol, 1.0 eq.) obtained in the previous step was dissolved in dichloromethane (100 mL), then a 4 mol/L HCl/1,4-dioxane solution (45 mL, 5.0 eq.) was added, and the reaction solution was stirred at room temperature for 30 min. After the reaction was completed as detected by TLC, the reaction solution was poured into water, followed by liquid separation. Sodium bicarbonate solid was added to the aqueous phase to adjust the pH to 8, followed by concentration. The crude product was dissolved in DCM, filtered under vacuum, and concentrated to give the product (8 g, yield: 84.4%).
(R)-4-chloro-N-(piperidin-3-yl)-5,6,7,8-tetrahydrophthalazin-1-amine (8 g, 30 mmol, 1.0 eq.) obtained in the previous step was dissolved in methanol (100 mL), then an aqueous formaldehyde solution (37%) (2.43 g, 30 mmol, 1.0 eq.) was added, and the reaction solution was stirred at room temperature for 30 min. NaBH3CN (2.45 g, 39 mmol, 1.3 eq.) was added, and the resulting solution was stirred at room temperature for 1 h. After the reaction was completed as detected by TLC, the reaction solution was concentrated, dissolved in DCM (100 mL), and washed with a saturated aqueous sodium bicarbonate solution (100 mL×2). The organic phase was dried and filtered under vacuum, the filtrate was concentrated, and the residue was purified by silica gel column chromatography (dichloromethane:methanol=50:1 to 30:1) to give the product (5 g, yield: 59.3%).
(R)-4-chloro-N-(1-methylpiperidin-3-yl)-5,6,7,8-tetrahydrophthalazin-1-amine (500 mg, 1.78 mmol, 1.0 eq.) obtained in the previous step was dissolved in 1,4-dioxane (4 mL) and water (2 mL), then (4-fluoro-2-hydroxyphenyl)boronic acid (333 mg, 2.14 mmol, 1.2 eq.), sodium bicarbonate (300 mg, 3.56 mmol, 2.0 eq.) and Pd(dppf)Cl2 (130 mg, 0.178 mmol, 0.1 eq.) were added successively, and the reaction solution was stirred at 110° C. overnight in nitrogen atmosphere. After the reaction was completed as detected by TLC, the reaction solution was concentrated and extracted with dichloromethane (5 mL×3). The organic phase was dried and filtered under vacuum, the filtrate was concentrated, and the crude product was purified by silica gel column chromatography (dichloromethane:methanol=20:1 to 10:1) to give a crude product (600 mg), and the crude product (200 mg) was purified by preparative thin-layer chromatography to give the product (150 mg, yield: 71.0%).
1H-NMR (400 MHz, DMSO-d6) δ (ppm): 10.26 (s, 1H), 7.13 (t, J=7.96 Hz, 1H), 6.74-6.69 (m, 2H), 5.69 (s, 1H), 4.34 (s, 1H), 3.35 (s, 1H), 3.08 (s, 1H), 2.79 (s, 1H), 2.38-2.31 (m, 8H), 1.87-1.52 (m, 8H).
Molecular formula: C20H25FN4O Precise molecular weight: 356.20 LC-MS (m/z): 357.12 [M+H]+.
4-Bromo-2-fluoro-1-methylbenzene (2 g, 10.58 mmol, 1.0 eq.) was dissolved in dry tetrahydrofuran (15 mL), and the reaction solution was cooled to −70° C., followed by dropwise addition of a 2 mol/L solution of LDA in n-hexane (6 mL, 12 mmol, 1.13 eq.). After stirring for 2 h, triisopropyl borate (2.07 g, 11 mmol, 1.05 eq.) was added dropwise, with stirring for 20 min, then a 2 mol/L solution of LDA in n-hexane (2 mL, 4 mmol, 0.38 eq.) was added dropwise, with stirring for 20 min, and then triisopropyl borate (0.6 g, 3.2 mmol, 0.3 eq.) was added dropwise, with stirring for 1 h. The resulting solution was heated to 0° C., a solution of acetic acid (2 mL) in water (15 mL) was poured into the reaction solution, an aqueous hydrogen peroxide solution (30%, 5 mL) was added dropwise to the system, and the reaction was carried out overnight at room temperature. After the reaction was completed as detected by TLC, liquid separation was carried out, and the pH of an aqueous phase was adjusted to 6, followed by extraction with ethyl acetate (15 mL). The organic phases were combined, washed successively with a saturated aqueous sodium sulfite solution (15 mL) and a 1 mol/L aqueous hydrochloric acid solution (15 mL), and extracted with a 1 mol/L aqueous sodium hydroxide solution (15 mL×2). The pH of the aqueous phase was adjusted to 6, followed by extraction with dichloromethane (15 mL×2). The organic phases were combined, dried, and concentrated to give the product (2.1 g, yield: 96.8%).
6-Bromo-2-fluoro-3-methylphenol (1 g, 4.88 mmol, 1.0 eq.) obtained in the previous step was dissolved in dry tetrahydrofuran (6 mL), 60% (mass fraction) NaH (293 mg, 7.32 mmol, 1.5 eq.) was added portionwise under an ice-water bath, with stirring for 30 min, followed by addition of chloromethyl ethyl ether (599 mg, 634 mmol, 1.3 eq.), and the reaction solution was stirred at room temperature overnight. After the reaction was completed as detected by TLC, water was added to the reaction solution to quench the reaction, followed by concentration, and then water (5 mL) and ethyl acetate (10 mL) were added, followed by liquid separation. The organic phase was dried and concentrated to give the product (1.1 g, yield: 85.7%).
1-Bromo-2-(ethoxymethoxy)-3-fluoro-4-methylbenzene (526 mg, 2 mmol, 1.0 eq.) obtained in the previous step was dissolved in dry tetrahydrofuran (6 mL), and the reaction solution was cooled to −70° C., followed by dropwise addition of a 1.6 mol/L solution of n-butyllithium in n-hexane (1.9 mL, 3 mmol, 1.5 eq.). After stirring for 2 h, triisopropyl borate (489 mg, 2.6 mmol, 1.3 eq.) was added dropwise, and the resulting solution was stirred at room temperature for 3 h. After the reaction was completed as detected by TLC, water was added to the reaction solution to quench the reaction, followed by concentration. A 2 mol/L aqueous hydrochloric acid solution was added to adjust the pH to 6, followed by addition of water (5 mL) and ethyl acetate (10 mL), and liquid separation. The organic phase was dried and concentrated to give the product (450 mg, yield: 98.7%).
(R)-4-chloro-N-(1-methylpiperidin-3-yl)-5,6,7,8-tetrahydrophthalazin-1-amine (250 mg, 0.89 mmol, 1.0 eq.) was dissolved in 1,4-dioxane (2 mL) and water (1 mL), then (2-(ethoxymethoxy)-3-fluoro-4-methylphenyl)boronic acid (406 mg, 1.78 mmol, 2 eq.), sodium bicarbonate (150 mg, 1.78 mmol, 2.0 eq.) and Pd(dppf)Cl2 (65 mg, 0.089 mmol, 0.1 eq.) were added successively, and the reaction solution was stirred at 110° C. overnight in nitrogen atmosphere. After the reaction was completed as detected by TLC, the reaction solution was concentrated, followed by addition of water (5 mL) and extraction with dichloromethane (5 mL×3). The organic phase was dried and filtered under vacuum, the filtrate was concentrated, and the crude product was purified by silica gel column chromatography (dichloromethane:methanol=20:1 to 10:1) to give the product (300 mg, yield: 78.7%).
(R)-4-(2-(ethoxymethoxy)-3-fluoro-4-methylphenyl)-N-(1-methylpiperidin-3-yl)-5,6,7,8-tetrahydrophthalazin-1-amine (300 mg, 0.7 mmol, 1.0 eq.) obtained in the previous step was dissolved in dichloromethane (3 mL), then a 4 mol/L HCl/1,4-dioxane solution (0.88 mL, 5.0 eq.) was added, and the reaction solution was stirred at room temperature for 3 h. After the reaction was completed as detected by TLC, the reaction solution was poured into water, and the pH was adjusted to 8 with sodium bicarbonate solid, followed by liquid separation and extraction with dichloromethane (5 mL×2). The organic phase was dried, filtered under vacuum, and concentrated, and the crude product was purified by preparative thin-layer chromatography to give the product (200 mg, yield: 77.1%).
1H-NMR (400 MHz, DMSO-d6) δ (ppm): 9.84 (s, 1H), 6.85 (d, J=7.84 Hz, 1H), 6.75 (t, J=7.4 Hz, 1H), 5.72 (s, 1H), 4.33 (s, 1H), 3.35 (s, 1H), 3.06 (s, 1H), 2.77 (s, 1H), 2.38-2.33 (m, 7H), 2.26 (s, 3H), 1.87-1.52 (m, 9H).
Molecular formula: C21H27FN4O Precise molecular weight: 370.22 LC-MS (m/z): 371.20 [M+H]+.
(R)-3-(ethoxymethoxy)-4-(4-(piperidin-3-ylamino)-5,6,7,8-tetrahydrophthalazin-1-yl)benzonitrile (370.0 mg, 0.90 mmol, 1.0 eq.), (1-methoxycyclopropoxy)trimethylsilane (627.5 mg, 3.60 mmol, 4.0 eq.), acetic acid (1.0 mL), cesium fluoride (205.0 mg, 1.35 mmol, 1.5 eq.) and sodium cyanoborohydride (282.7 mg, 4.50 mmol, 5.0 eq.) were added to methanol (15.0 mL), and the reaction solution was reacted at 60° C. for 12 h. After the reaction was completed as monitored by TLC, a saturated aqueous sodium bicarbonate solution (50.0 mL) was added, followed by extraction with dichloromethane (50.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (dichloromethane:methanol=40:1 to 30:1) to give the product (310.0 mg, yield: 77.1%).
(R)-4-(4-((1-cyclopropylpiperidin-3-yl)amino)-5,6,7,8-tetrahydrophthalazin-1-yl)-3-(ethoxymethoxy)benzonitrile (280.0 mg, 0.62 mmol, 1.0 eq.) was added to ethanol (3.0 mL) and a hydrogen chloride-ethanol solution (2.0 mol/L) (5.0 mL), and the reaction solution was reacted at room temperature for 4 h. After the reaction was completed as monitored by TLC, the pH was adjusted to 7-8 with a saturated aqueous sodium bicarbonate solution, followed by extraction with dichloromethane (50.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and the crude product was purified by preparative thin-layer chromatography to give the product (130.0 mg, yield: 53.4%).
1HNMR (400 MHz, DMSO-d6) δ (ppm): 10.50 (s, 1H), 7.33 (s, 2H), 7.24 (s, 1H), 5.61-5.59 (d, J=8 Hz, 1H), 4.19-4.17 (m, 1H), 3.10-3.08 (d, J=8 Hz, 1H), 2.80-2.78 (d, J=8 Hz, 1H), 2.39-2.16 (m, 5H), 1.84-1.45 (m, 9H), 0.87-0.84 (m, 1H), 0.41-0.37 (m, 2H), 0.34-0.30 (m, 2H).
Molecular formula: C23H27N5O Precise molecular weight: 389.22 LC-MS (Pos, m/z)=390.15 [M+H]+.
(R)-4-chloro-N-(piperidin-3-yl)-5,6,7,8-tetrahydrophthalazin-1-amine (504.0 mg, 1.88 mmol, 1.0 eq.), 2-(2-(ethoxymethoxy)-4-methoxyphenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (1.74 g, 5.66 mmol, 3.0 eq.), sodium bicarbonate (631.7 mg, 7.52 mmol, 4.0 eq.) and Pd(dppf)Cl2 (137.5 mg, 0.18 mmol, 0.1 eq.) were added to 1,4-dioxane (15.0 mL) and water (7.0 mL), and the reaction solution was reacted at 110° C. for 2 h in nitrogen atmosphere. After the reaction was completed as monitored by TLC, the system was cooled to room temperature, and water (50.0 mL) was added, followed by extraction with ethyl acetate (100.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (100-200 mesh silica gel, dichloromethane:methanol=40:1 to 10:1) to give the product (503.0 mg, yield: 64.8%).
(R)-4-(2-(ethoxymethoxy)-4-methoxyphenyl)-N-(piperidin-3-yl)-5,6,7,8-tetrahydrophthalazin-1-amine (342.0 mg, 0.82 mmol, 1.0 eq.) was added to methanol (12.0 mL), then an aqueous formaldehyde solution (mass fraction: 37%, 80.7 mg, 0.99 mmol, 1.2 eq.) was added, and the reaction solution was reacted at room temperature for 0.5 h. Sodium cyanoborohydride (67.7 mg, 1.07 mmol, 1.3 eq.) was added, and the resulting solution was reacted for 1 h. After the reaction was completed as monitored by TLC, a saturated aqueous sodium bicarbonate solution was added to the system to adjust the pH to 7-8, followed by extraction with dichloromethane (50.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (100-200 mesh silica gel, dichloromethane:methanol=40:1 to 20:1) to give the product (270.0 mg, yield: 76.3%).
(R)-4-(2-(ethoxymethoxy)-4-methoxyphenyl)-N-(1-methylpiperidin-3-yl)-5,6,7,8-tetrahydrophthalazin-1-amine (270.0 mg, 0.63 mmol, 1.0 eq.) and a hydrogen chloride-ethanol solution (2.0 mol/L, 10.0 mL) were added to ethanol (10.0 mL), and the reaction solution was reacted at room temperature for 2 h. After the reaction was completed as monitored by TLC, a saturated aqueous sodium bicarbonate solution was added to the system to adjust the pH to 7-8, followed by extraction with dichloromethane (50.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and the crude product was purified by preparative thin-layer chromatography to give the product (130.0 mg, yield: 56.0%).
1HNMR (400 MHz, DMSO-d6) δ (ppm): 9.94 (s, 1H), 7.05-7.03 (d, J=8 Hz, 1H), 6.46-6.44 (m, 2H), 5.59-5.58 (d, J=4 Hz, 1H), 4.28 (s, 1H), 3.74 (s, 3H), 3.00 (s, 1H), 2.70 (s, 1H), 2.37-2.34 (m, 4H), 2.28 (s, 3H), 2.09 (s, 1H), 1.85-1.82 (m, 1H), 1.75-1.73 (m, 3H), 1.59-1.47 (m, 5H).
Molecular formula: C21H28N4O2 Precise molecular weight: 368.22 LC-MS (Pos, m/z)=369.18 [M+H]+.
(R)-4-chloro-N-(1-methylpiperidin-3-yl)-5,6,7,8-tetrahydrophthalazin-1-amine (500.0 mg, 1.78 mmol, 1.0 eq.), 3-(ethoxymethoxy)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzaldehyde (817.7 mg, 2.67 mmol, 1.5 eq.), sodium bicarbonate (299.0 mg, 3.56 mmol, 2.0 eq.) and Pd(dppf)Cl2 (65.1 mg, 0.08 mmol, 0.05 eq.) were added to 1,4-dioxane (10.0 mL) and water (5.0 mL), and the reaction solution was reacted at 110° C. for 4 h in nitrogen atmosphere. After the reaction was completed as monitored by TLC, water (100.0 mL) was added, and ethyl acetate (200.0 mL) was added for extraction. The organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (100-200 mesh silica gel, dichloromethane:methanol=40:1 to 10:1) to give the product (430.0 mg, yield: 56.9%).
(R)-3-(ethoxymethoxy)-4-(4-((1-methylpiperidin-3-yl)amino)-5,6,7,8-tetrahydrophthalein-1-yl)benzaldehyde (430.0 mg, 1.01 mmol, 1.0 eq.) was added to dichloromethane (10.0 mL), then diethylaminosulfur trifluoride (244.8 mg, 1.51 mmol, 1.5 eq.) was added dropwise under an ice bath, and the reaction solution was warmed to room temperature and stirred for 12 h. After the reaction was completed as monitored by TLC, a saturated aqueous sodium carbonate solution was added to adjust the pH to 8-9 under an ice bath, followed by extraction with dichloromethane (100.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and the crude product was purified by preparative thin-layer chromatography to give the product (220.0 mg, yield: 48.7%).
(R)-4-(4-(difluoromethyl)-2-(ethoxymethoxy)phenyl)-N-(1-methylpiperidin-3-yl)-5,6,7,8-tetrahydrophthalazin-1-amine (180.0 mg, 0.40 mmol, 1.0 eq.) and a hydrogen chloride-1,4-dioxane solution (4.0 mol/L, 3.0 mL) were added to dichloromethane (1.0 mL), and the reaction solution was stirred at room temperature for 2 h. After the reaction was completed as monitored by TLC, a saturated aqueous sodium bicarbonate solution was added to the system to adjust the pH to 7-8, followed by extraction with dichloromethane (100.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and the crude product was purified by preparative thin-layer chromatography to give the product (30.0 mg, yield: 19.2%).
1HNMR (400 MHz, DMSO-d6) δ (ppm): 10.16 (s, 1H), 7.25-7.23 (d, J=8 Hz, 1H), 7.14 (s, 0.20H), 7.06-7.03 (m, 2H), 7.00 (s, 0.43H), 6.86 (s, 0.24H), 5.54-5.52 (d, J=8 Hz, 1H), 4.28-4.22 (m, 1H), 2.90-2.88 (d, J=8 Hz, 1H), 2.59-2.57 (d, J=8 Hz, 1H), 2.38-2.31 (m, 4H), 2.18 (s, 3H), 1.99-1.91 (m, 2H), 1.84-1.40 (m, 8H).
Molecular formula: C21H26F2N4O Precise molecular weight: 388.21 LC-MS (Pos, m/z)=389.14 [M+H]+.
Dimethyl acetylenedicarboxylate (10.0 g, 70.4 mmol, 1.0 eq.) and 1,3-cyclohexadiene (6.77 g, 84.5 mmol, 1.2 eq.) were dissolved in THF (50 mL), and the reaction solution was heated to 65° C. in nitrogen atmosphere and reacted for 20 h, then cooled to room temperature, and concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (PE:EA=50:1) to give the product (12.5 g, yield: 79.9%).
Dimethyl bicyclo[2.2.2]octa-2,5-diene-2,3-dicarboxylate (12.0 g, 54.0 mmol, 1.0 eq.) was dissolved in EtOH (50 mL), followed by addition of hydrazine hydrate (85%, 32.0 g, 540 mmol, 10.0 eq.), and the reaction solution was reacted at room temperature for 30 h, and concentrated to give the product (10.0 g, yield: 97.4%).
5,8-Dihydro-5,8-ethanophthalazine-1,4-diol (10.0 g, 52.6 mmol, 1.0 eq.) was added to ACN (50 mL), then POCl3 (50 mL) was added, and the reaction solution was heated to 100° C. under an oil bath and reacted for 5 h, then cooled to room temperature, concentrated, washed with water (50 mL), and filtered under vacuum to give the product (2.60 g, yield: 21.8%).
1,4-Dichloro-5,8-dihydro-5,8-ethanophthalazine (2.60 g, 11.4 mmol, 1.0 eq.), tert-butyl (R)-3-aminopiperidine-1-carboxylate (2.52 g, 12.6 mmol, 1.1 eq.), Pd2(dba)3 (1.05 g, 1.14 mmol, 0.1 eq.), BINAP (1.42 g, 2.28 mmol, 0.2 eq.) and Cs2CO3 (7.46 g, 22.4 mmol, 2.0 eq.) were added successively to toluene (50 mL), and the reaction solution was heated to 90° C. in nitrogen atmosphere and reacted for 2 h, and then cooled to room temperature. Water was added to quench the reaction, followed by extraction with EA (30 mL×2). The organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and the crude product was separated by silica gel column chromatography (DCM:MeOH=100:1 to 50:1) to give the product (2.70 g, yield: 60.3%).
3-(Ethoxymethoxy)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzonitrile (349 mg, 1.15 mmol, 1.5 eq.), (R)-4-chloro-N-(1-(tetrahydro-2H-pyran-4-yl)piperidin-3-yl)phthalazin-1-amine (300 mg, 0.767 mmol, 1.0 eq.), Pd(dppf)Cl2 (28.1 mg, 0.0384 mmol, 0.05 eq.) and NaHCO3 (128 mg, 1.53 mmol, 2.0 eq.) were added successively to 1,4-dioxane (6 mL) and water (3 mL), and the reaction solution was heated to 110° C. in nitrogen atmosphere and reacted for 2 h, and then cooled to room temperature. Water was added to quench the reaction, followed by extraction with EA (20 mL×2). The organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and the crude product was separated by silica gel column chromatography (DCM:MeOH=50:1) to give the product (280 mg, yield: 68.6%).
Tert-butyl (3R)-3-((4-(4-cyano-2-(ethoxymethoxy)phenyl)-5,8-dihydro-5,8-ethanophthalazin-1-yl)amino)piperidine-1-carboxylate (270 mg, 0.508 mmol, 1.0 eq.) was dissolved in DCM (3 mL), then a solution of hydrogen chloride in 1,4-dioxane (4 mol/L, 1.27 mL, 5.08 mmol, 10.0 eq.) was added dropwise, and the reaction solution was stirred at room temperature for 2 h. Water (20 mL) was added to quench the reaction, followed by washing with DCM (15 mL×2), and a saturated aqueous NaHCO3 solution was added to the aqueous phase to adjust the pH to 8, followed by extraction with DCM (20 mL×3). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to give the product (168 mg, yield: 88.6%).
3-Hydroxy-4-(4-(((R)-piperidin-3-yl)amino)-5,8-dihydro-5,8-ethanophthalazin-1-yl)benzonitrile (168 mg, 0.434 mmol, 1.0 eq.) was dissolved in MeOH (4 mL), then an aqueous formaldehyde solution (37%, 42.3 mg, 0.521 mmol, 1.2 eq.) was added, and the reaction solution was stirred at room temperature for 20 min. NaBH3CN (32.7 mg, 0.521 mmol, 1.2 eq.) was then added, and the resulting solution was reacted for 1 h, concentrated, and dissolved in DCM. A saturated aqueous NaHCO3 solution was added to adjust the pH to 8, followed by extraction with DCM (20 mL×2). The organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and the crude product was separated by preparative thin-layer chromatography (DCM:MeOH=10:1) to give the product (36.8 mg, yield: 21.1%).
1HNMR (400 MHz, DMSO-d6) δ (ppm): 10.75 (s, 1H), 7.44 (d, J=8.0 Hz, 1H), 7.39-7.36 (m, 1H), 7.33 (s, 1H), 6.49-6.41 (m, 3H), 4.48 (s, 1H), 4.29 (s, 1H), 3.76 (d, J=5.6 Hz, 1H), 3.15 (s, 2H), 2.83 (s, 1H), 2.34 (s, 3H), 2.08 (s, 1H), 1.92 (s, 1H), 1.78 (s, 1H), 1.64-1.61 (m, 1H), 1.47-1.39 (m, 4H), 1.34-1.31 (m, 1H).
Molecular formula: C23H25N5O Precise molecular weight: 387.21 LC-MS (Pos, m/z)=388.13 [M+H]+.
2-(4-Cyclopropyl-2-methoxyphenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (313 mg, 1.14 mmol, 1.1 eq.), (R)-4-chloro-N-(1-methylpiperidin-3-yl)-5,6,7,8-tetrahydro-5,8-ethanophthalazin-1-amine (320 mg, 1.04 mmol, 1.0 eq.), Pd(dppf)Cl2 (76.1 mg, 0.104 mmol, 0.1 eq.) and NaHCO3 (175 mg, 2.08 mmol, 2.0 eq.) were added successively to 1,4-dioxane (6 mL) and H2O (3 mL), and the reaction solution was heated to 110° C. in nitrogen atmosphere and reacted for 2 h, and then cooled to room temperature. Water was added to quench the reaction, followed by extraction with EA (20 mL×3). The organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and the crude product was separated by silica gel column chromatography (DCM:MeOH=50:1 to 10:1) to give the product (360 mg, yield: 82.5%).
((R)-4-(4-cyclopropyl-2-methoxyphenyl)-N-(1-methylpiperidin-3-yl)-5,6,7,8-tetrahydro-5,8-ethanophthalazin-1-amine (340 mg, 0.812 mmol, 1.0 eq.) was dissolved in DCM (10 mL), and the reaction solution was cooled to −10° C. BBr3 (611 mg, 2.44 mmol, 3.0 eq.) was added dropwise, and the reaction was continued for 2 h. MeOH was added to quench the reaction, and a saturated aqueous NaHCO3 solution was added to adjust the pH to 8, followed by extraction with DCM (20 mL×2). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was separated by preparative thin-layer chromatography (DCM:MeOH=8:1) to give the product (38.0 mg, yield: 11.6%).
1HNMR (400 MHz, CDCl3) δ(ppm): 11.75 (s, 1H), 7.19 (d, J=8.0 Hz, 1H), 6.79 (d, J=1.6 Hz, 1H), 6.69-6.67 (m, 1H), 5.24 (s, 1H), 4.48 (s, 1H), 3.59 (s, 1H), 5.10 (s, 1H), 2.66 (s, 1H), 2.50 (s, 1H), 2.29 (s, 3H), 2.14 (m, 1H), 1.93-1.82 (m, 6H), 1.76-1.70 (m, 2H), 1.46-1.39 (m, 4H), 1.02-1.97 (m, 2H), 0.91-0.88 (m, 1H), 0.79-0.85 (m, 2H).
Molecular formula: C25H32N4O Precise molecular weight: 404.26 LC-MS (Pos, m/z)=405.17 [M+H]+.
1,4-Dichloro-5,6,7,8-tetrahydrophthalazine (518.0 mg, 2.55 mmol, 1.0 eq.), (2-(ethoxymethoxy)-4-(trifluoromethyl)phenyl) boronic acid (740.7 mg, 2.80 mmol, 1.1 eq.), sodium bicarbonate (428.0 mg, 5.10 mmol, 2.0 eq.) and Pd(dppf)Cl2 (93.2 mg, 0.12 mmol, 0.05 eq.) were added to a mixed solution of 1,4-dioxane (10.0 mL) and water (4.0 mL), and the reaction solution was reacted at 110° C. for 3 h in nitrogen atmosphere. After the reaction was completed as monitored by TLC, water (100.0 mL) was added, and ethyl acetate (100.0 mL) was added for extraction. The organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=20:1 to 8:1) to give the product (450.0 mg, yield: 45.6%).
1-Chloro-4-(2-(ethoxymethoxy)-4-(trifluoromethyl)phenyl)-5,6,7,8-tetrahydrophthalazine (400.0 mg, 1.03 mmol, 1.0 eq.), (cis)-3-amino-1-methylcyclobutan-1-ol hydrochloride (213.4 mg, 1.55 mmol, 1.5 eq.), cesium carbonate (673.7 mg, 2.06 mmol, 2.0 ethoxyq.), Pd2(dba)3 (94.6 mg, 0.10 mmol, 0.1 eq.) and 1,1′-binaphthyl-2,2′-bisdiphenylphosphine (128.7 mg, 0.20 mmol, 0.2 eq.) were added to 1,4-dioxane (15.0 mL), and the reaction solution was reacted at 90° C. for 12 h in nitrogen atmosphere. After the reaction was completed as monitored by TLC, water (100.0 mL) was added, and ethyl acetate (100.0 mL) was added for extraction. The organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (specification of silica gel: 100-200 mesh, dichloromethane:methanol=80:1 to 50:1) to give the product (380.0 mg, yield: 81.5%).
(Cis)-3-((4-(2-(ethoxymethoxy)-4-(trifluoromethyl)phenyl)-5,6,7,8-tetrahydrophthalazin-1-yl)amino)-1-methylcyclobutan-1-ol (380.0 mg, 0.84 mmol, 1.0 eq.) was added to dichloromethane (2.0 mL), then a hydrogen chloride-1,4-dioxane solution (4.0 mol/L, 6.0 mL) was added dropwise, and the reaction solution was reacted at room temperature for 2 h. After the reaction was completed as monitored by TLC, a saturated aqueous sodium bicarbonate solution was added to the system to adjust the pH to 7-8, followed by extraction with dichloromethane (100.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to give the product (222.0 mg, yield: 67.2%).
1HNMR (400 MHz, DMSO-d6) δ (ppm): 11.14 (s, 1H), 8.28 (s, 1H), 7.48-7.46 (d, J=8 Hz, 1H), 7.42 (s, 1H), 7.30-7.28 (d, J=8 Hz, 1H), 5.16 (s, 1H), 4.05-4.00 (m, 1H), 3.57 (s, 1H), 2.61-2.59 (m, 2H), 2.42-2.41 (m, 2H), 2.30-2.25 (m, 2H), 1.79-1.65 (m, 4H), 1.30 (s, 3H).
Molecular formula: C20H22F3N3O2 Precise molecular weight: 393.17 LC-MS (m/z)=394.11 [M+H]+.
(Cis)-3-((4-chloro-5,6,7,8-tetrahydrophthalazin-1-yl)amino)-1-methylcyclobutan-1-ol (180.0 mg, 0.67 mmol, 1.0 eq.), 3-(ethoxymethoxy)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzonitrile (305.5 mg, 1.00 mmol, 1.5 eq.), sodium bicarbonate (112.9 mg, 1.34 mmol, 2.0 eq.) and Pd(dppf)Cl2 (24.5 mg, 0.03 mmol, 0.05 eq.) were added to a mixed solution of 1,4-dioxane (10.0 mL) and water (5.0 mL), and the reaction solution was reacted at 110° C. for 4 h in nitrogen atmosphere. After the reaction was completed as monitored by TLC, water (100.0 mL) was added, and ethyl acetate (100.0 mL) was added for extraction. The organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (dichloromethane:methanol=80:1 to 30:1) to give the product (130.0 mg, yield: 47.3%).
3-(Ethoxymethoxy)-4-(4-(((cis)-3-hydroxy-3-methylcyclobutyl)amino)-5,6,7,8-tetrahydrophthalazin-1-yl)benzonitrile (130.0 mg, 0.31 mmol, 1.0 eq.) was added to dichloromethane (2.0 mL), then a hydrogen chloride-1,4-dioxane solution (4.0 mol/L, 4.0 mL) was added dropwise, and the reaction solution was reacted at room temperature for 2 h. After the reaction was completed as monitored by TLC, a saturated aqueous sodium bicarbonate solution was added to the system to adjust the pH to 7-8, followed by extraction with dichloromethane (50.0 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to give the product (100.0 mg, yield: 90.0%).
1HNMR (400 MHz, DMSO-d6) δ (ppm): 10.53 (s, 1H), 7.32 (s, 2H), 7.24 (s, 1H), 6.17-6.16 (t, J=4 Hz, 1H), 4.93 (s, 1H), 4.11-4.06 (m, 1H), 2.39-2.29 (m, 6H), 2.09-2.05 (m, 2H), 1.75-1.59 (m, 4H), 1.29 (s, 3H).
Molecular formula: C20H22N4O2 Precise molecular weight: 350.17 LC-MS (m/z)=351.08 [M+H]+.
NaH (6.64 g, 166 mmol, 1.2 eq.) was suspended in dichloromethane (600 mL), then a solution of methyl 2-oxocycloheptane-1-carboxylate (23.5 g, 138 mmol, 1.0 eq.) in dichloromethane (50 mL) was added dropwise at 0° C., and the reaction solution was stirred at 0° C. for 30 min. A solution of trifluoromethanesulfonic anhydride (47 g, 166 mmol, 1.2 eq.) in dichloromethane (100 mL) was added dropwise, the resulting solution was reacted at 25° C. for 30 min, followed by supplementation with dichloromethane (450 mL), and then the reaction was carried out at 25° C. for 16 h. Water was added dropwise to the reaction solution to quench the reaction, followed by liquid separation. The organic phase was concentrated, and the crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=30:1) to give a crude product (35 g, yield: 83%).
Sodium formate (4.1 g, 59.60 mmol, 3.0 eq.), DIEA (5.12 g, 39.74 mmol, 2.0 eq.) and acetic anhydride (4 g, 39.74 mmol, 2.0 eq.) were dissolved in DMF (20 mL) with stirring for 30 min, then DMF (40 mL), methyl 2-(((trifluoromethyl)sulfonyl)oxy)cyclohept-1-ene-1-carboxylate (6 g, 19.87 mmol, 1.0 eq.), palladium acetate (220 mg, 0.99 mmol, 0.05 eq.) and lithium chloride (2.5 g, 59.60 mmol, 3.0 eq.) were added, and the reaction solution was reacted at 25° C. for 12 h. After the materials reacted completely as monitored by TLC, the reaction solution was poured into a solution of ethyl acetate and 2 mol/L hydrochloric acid, and after stirring for 5 min, liquid separation was carried out. The organic phase was dried and concentrated, and the crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=5:1) to give a crude product (3 g, yield: 91%).
5,6,7,8-Tetrahydro-1H-cyclohepta[c]furan-1,3(4H)-dione (3 g, 18.07 mmol, 1.0 eq.) was added to acetic acid (18 mL) and water (20 mL), then sodium acetate (2.2 g, 27.1 mmol, 1.5 eq.) and hydrazine hydrate (1.35 g, 27.1 mmol, 1.5 eq.) were added, and the reaction solution was reacted at 110° C. for 18 h. The solvent was removed under reduced pressure, the crude product was slurried with MTBE and filtered under vacuum, and the filter cake was washed with ethyl acetate to give a crude product (3.5 g).
2,3,6,7,8,9-Hexahydro-1H-cyclohepta[d]pyridazine-1,4(5H)-dione (3.5 g, 19.4 mmol, 1.0 eq.) and DIPEA (2.5 g, 19.4 mmol, 1.0 eq.) were added to phosphorus oxychloride (20 mL), and the reaction solution was reacted at 110° C. for 40 min. The solvent was removed under reduced pressure, the crude product was diluted with dichloromethane, and a saturated aqueous sodium bicarbonate solution was added to adjust the pH to alkalinity, followed by liquid separation. The organic phase was dried and concentrated, and the crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=5:1) to give the product (800 mg, two-step yield: 20%).
1,4-Dichloro-6,7,8,9-tetrahydro-5H-cyclohepta[d]pyridazine (800 mg, 3.69 mmol, 1.0 eq.) was added to dioxane (15 mL), then Pd2(dba)3 (337 mg, 0.37 mmol, 0.1 eq.), BINAP (458 mg, 0.74 mmol, 0.2 eq.), cesium carbonate (3.6 g, 11.06 mmol, 3.0 eq.) and tert-butyl (R)-3-aminopiperidine-1-carboxylate (737 mg, 3.69 mmol, 1.0 eq.) were added, and the reaction solution was reacted at 110° C. for 3 h in nitrogen atmosphere. After there remained a small number of materials as detected by TLC, the reaction solution was filtered through celite under vacuum, the filtrate was concentrated, and the crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=5:1 to 1:1) to give the product (1 g, yield: 71%).
Tert-butyl (R)-3-((4-chloro-6,7,8,9-tetrahydro-5H-cyclohepta[d]pyridazin-1-yl)amino)piperidine-1-carboxylate (600 mg, 1.58 mmol, 1.0 eq.) was added to dioxane (10 mL) and water (2 mL), then Pd(PPh3)4 (182 mg, 0.16 mmol, 0.1 eq.), potassium carbonate (653 mg, 4.74 mmol, 2.5 eq.) and (2-methoxy-4-methylphenyl)boronic acid (288 mg, 1.74 mmol, 1.1 eq.) were added, and the reaction solution was reacted at 90° C. for 5 h in nitrogen atmosphere. After the materials reacted completely as monitored by TLC, the reaction solution was filtered through celite under vacuum, the filtrate was concentrated, and the crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=3:1 to 1:1) to give the product (660 mg, yield: 90%).
Tert-butyl (R)-3-((4-(2-methoxy-4-methylphenyl)-6,7,8,9-tetrahydro-5H-cyclohepta[d]pyridazin-1-yl)amino)piperidine-1-carboxylate (660 mg, 1.43 mmol, 1.0 eq.) was dissolved in dichloromethane (12 mL), then boron tribromide (1.8 g, 7.08 mmol, 5.0 eq.) was added, and the reaction solution was reacted at 25° C. for 1 h. After the materials reacted completely as monitored by TLC, water was added to the reaction solution at 0° C. to quench the reaction, followed by liquid separation. A saturated aqueous sodium bicarbonate solution was added to the aqueous phase to adjust the pH to alkalinity, followed by extraction with dichloromethane. The organic phase was dried and concentrated to give the product (380 mg, yield: 76%).
(R)-5-methyl-2-(4-(piperidin-3-ylamino)-6,7,8,9-tetrahydro-5H-cyclohepta[d]pyridazin-1-yl)phenol (380 mg, 1.08 mmol, 1.0 eq.) was dissolved in methanol (8 mL), then an aqueous formaldehyde solution (37%) (87 mg, 1.08 mmol, 1.0 eq.) and sodium cyanoborohydride (75 mg, 1.19 mmol, 1.1 eq.) were added, and the reaction solution was reacted at 25° C. for 20 min. After the materials reacted completely as monitored by TLC, the reaction solution was concentrated, and dichloromethane and saturated sodium bicarbonate were added, followed by liquid separation. The organic phase was dried, concentrated, and purified by preparative thin-layer chromatography (dichloromethane:methanol=7:1) to give the product (212 mg, yield: 54%).
1H-NMR (400 MHz, DMSO-d6) δ (ppm): 9.49 (s, 1H), 7.99-7.97 (d, J=7.6 Hz, 1H), 6.73 (s, 1H), 6.68-6.66 (d, J=7.6 Hz, 1H), 6.29 (s, 1H), 4.50 (s, 1H), 3.68-3.51 (m, 1H), 3.45-3.43 (m, 1H), 3.26 (m, 1H), 2.74 (m, 6H), 2.48 (s, 1H), 2.27 (s, 3H), 2.08-1.89 (m, 2H), 1.77-1.76 (m, 4H), 1.52-1.46 (m, 5H).
Molecular formula: C22H30N4O Precise molecular weight: 366.24 LC-MS (m/z): 367.17 [M+H]+.
6-bromo-2,3-difluorophenol (1 g, 4.78 mmol, 1.0 eq.) was dissolved in anhydrous tetrahydrofuran (6 mL), 60% (mass fraction) NaH (287 mg, 7.17 mmol, 1.5 eq.) was added portionwise under an ice-water bath, with stirring for 30 min, followed by addition of chloromethyl ethyl ether (587 mg, 6.21 mmol, 1.3 eq.), and the reaction solution was stirred at room temperature for 2 h. After the reaction was completed as detected by TLC, water was added to the reaction solution to quench the reaction, followed by concentration, and then water (5 mL) and ethyl acetate (10 mL) were added, followed by liquid separation. The organic phase was washed successively with water (15 mL×2) and saturated brine (15 mL×2), dried, and concentrated to give the product (1.1 g, yield: 86.2%).
1-Bromo-2-(ethoxymethoxy)-3,4-difluorobenzene (1.06 g, 4 mmol, 1.0 eq.) was dissolved in anhydrous tetrahydrofuran (12 mL), and the reaction solution was cooled to −70° C. A 1.6 mol/L solution of n-butyllithium in n-hexane (3.8 mL, 6 mmol, 1.5 eq.) was added dropwise, with stirring for 2 h, then triisopropyl borate (978 mg, 5.2 mmol, 1.3 eq.) was added dropwise, and the reaction solution was stirred at room temperature for 3 h. After the reaction was completed as detected by TLC, water was added to the reaction solution to quench the reaction, followed by concentration. A 2 mol/L aqueous hydrochloric acid solution was added to adjust the pH to 6, followed by addition of water (5 mL) and ethyl acetate (10 mL), and liquid separation. The organic phase was dried and concentrated to give the product (750 mg, yield: 80.8%).
(R)-4-chloro-N-(1-methylpiperidin-3-yl)-5,6,7,8-tetrahydrophthalazin-1-amine (250 mg, 0.89 mmol, 1.0 eq.) was dissolved in 1,4-dioxane (2 mL) and water (1 mL), then (2-(ethoxymethoxy)-3,4-difluorophenyl)boronic acid (413 mg, 1.78 mmol, 2.0 eq.), sodium bicarbonate (150 mg, 1.78 mmol, 2.0 eq.) and Pd(dppf)Cl2 (65 mg, 0.089 mmol, 0.1 eq.) were added successively, and the reaction solution was stirred at 110° C. overnight in nitrogen atmosphere. After there remained a small number of materials as detected by TLC, the reaction solution was concentrated, followed by addition of water (5 mL) and extraction with dichloromethane (5 mL×3). The organic phase was dried and filtered under vacuum, the filtrate was concentrated, and the crude product was purified by silica gel column chromatography (dichloromethane:methanol=20:1 to 10:1) to give the product (80 mg, yield: 20.8%).
(R)-4-(2-(ethoxymethoxy)-3,4-difluorophenyl)-N-(1-methylpiperidin-3-yl)-5,6,7,8-tetrahydrophthalazin-1-amine (80 mg, 0.19 mmol, 1.0 eq.) was dissolved in dichloromethane (1 mL), then a 4 mol/L solution of hydrogen chloride in 1,4-dioxane (0.23 mL, 5.0 eq.) was added, and the reaction solution was stirred at room temperature for 3 h. After the reaction was completed as detected by TLC, the reaction solution was poured into water, and the pH was adjusted to 8 with sodium bicarbonate, followed by liquid separation and extraction with dichloromethane (5 mL×2). The organic phase was dried, filtered under vacuum, and concentrated, and the crude product was purified by preparative thin-layer chromatography to give the product (37 mg, yield: 53.4%).
1H-NMR (400 MHz, DMSO-d6) δ (ppm): 10.47 (s, 1H), 7.00-6.89 (m, 2H), 5.82 (s, 1H), 4.38 (s, 1H), 3.15 (s, 1H), 2.87 (s, 1H), 2.77 (s, 1H), 2.40-2.31 (m, 6H), 1.85-1.60 (m, 10H).
Molecular formula: C20H24F2N4O Precise molecular weight: 374.19 LC-MS (m/z): 375.26 [M+H]+.
1HNMR (300 MHz, DMSO-d6) δ (ppm): 11.95 (s, 1H), 8.50 (s, 1H), 7.89-7.77 (m, 2H), 7.60-7.52 (m, 3H), 6.19 (d, J=6.9 Hz, 1H), 4.74 (s, 1H), 3.63-3.59 (m, 2H), 3.17 (s, 1H), 2.78 (s, 4H), 2.30 (s, 3H), 2.08 (s, 1H), 1.84 (s, 3H).
Molecular formula: C20H23N5O Precise molecular weight: 349.19 LC-MS (m/z): 350.28 [M+H]+.
1HNMR (300 MHz, DMSO-d6) δ (ppm): 9.37 (s, 1H), 8.41 (d, J=8.1 Hz, 1H), 7.86-7.74 (m, 2H), 7.54 (d, J=7.8 Hz, 1H), 7.14 (s, 1H), 7.04 (s, 1H), 6.80 (s, 1H), 4.49 (s, 1H), 3.21 (d, J=8.4 Hz, 1H), 2.84 (d, J=11.1 Hz, 1H), 2.35 (s, 3H), 2.23-2.18 (m, 8H), 2.02-1.97 (m, 1H), 7.84-1.80 (m, 1H), 1.72-1.49 (m, 2H).
Molecular formula: C22H26N4O Precise molecular weight: 362.21 LC-MS (m/z): 363.34 [M+H]+.
1HNMR (300 MHz, DMSO-d6) δ (ppm): 15.0 (s, 1H), 9.20 (d, J=1.2 Hz, 1H), 8.96 (s, 2H), 8.03 (s, 1H), 7.82 (d, J=8.4 Hz, 1H), 6.77 (d, J=5.7 Hz, 1H), 4.47 (s, 1H), 3.34 (d, J=8.4 Hz, 3H), 3.01 (d, J=11.7 Hz, 1H), 2.54 (s, 3H), 2.31 (s, 3H), 1.95-1.89 (m, 2H), 1.75-1.59 (m, 2H).
Molecular formula: C19H22N4O2 Precise molecular weight: 338.17 LC-MS (m/z): 339.15 [M+H]+.
1HNMR (300 MHz, DMSO-d6) δ (ppm): 9.00 (s, 1H), 8.58 (d, J=2.7 Hz, 1H), 8.05 (d, J=8.1 Hz, 2H), 7.42-7.38 (m, 2H), 4.62 (s, 1H), 3.18 (s, 2H), 2.79-2.73 (m, 1H), 2.68 (s, 3H), 2.04-1.99 (m, 3H), 1.81-1.77 (m, 2H).
Molecular formula: C19H19N5OS Precise molecular weight: 365.13 LC-MS (m/z): 366.14[M+H]+
1HNMR (300 MHz, DMSO-d6) δ (ppm): 8.30 (d, J=7.5 Hz, 1H), 7.94 (s, 1H), 7.90-7.83 (m, 2H), 7.73 (d, J=7.8 Hz, 1H), 6.51 (s, 1H), 4.63 (s, 1H), 3.78 (s, 3H), 3.43 (d, J=9.6 Hz, 1H), 3.09 (s, 2H), 2.90 (s, 3H), 2.47 (s, 3H), 2.15 (s, 2H), 1.91 (s, 2H).
Molecular formula: C20H23N5O Precise molecular weight: 349.19 LC-MS (m/z): 350.12 [M+H]+.
1HNMR (400 MHz, DMSO-d6) δ (ppm): 9.75 (s, 1H), 8.53 (s, 1H), 7.91-7.83 (m, 2H), 7.55 (d, J=8.0 Hz, 1H), 7.46-7.35 (m, 3H), 7.23 (d, J=7.6 Hz, 2H), 7.05 (s, 1H), 6.94-6.91 (m, 1H), 5.26 (d, J=3.2 Hz, 1H), 4.75-4.73 (m, 2H), 3.72 (s, 1H), 2.94 (s, 2H), 2.81 (s, 3H), 2.20-1.78 (m, 4H), 1.38 (d, J=6.4 Hz, 3H).
Molecular formula: C22H26N4O2 Precise molecular weight: 378.21 LC-MS (m/z): 379.16 [M+H]+.
1HNMR (400 MHz, DMSO-d6) δ (ppm): 9.61 (s, 1H), 8.44 (d, J=6.8 Hz, 1H), 7.88-7.78 (m, 2H), 7.53 (d, J=8.0 Hz, 1H), 7.28 (s, 1H), 7.21 (d, J=8.0 Hz, 1H), 7.15 (d, J=1.2 Hz, 1H), 7.03-7.01 (m, 1H), 5.07 (s, 1H), 4.57 (s, 1H), 3.02 (s, 2H), 2.01-1.89 (m, 3H), 1.74-1.63 (m, 3H), 1.47 (s, 6H).
Molecular formula: C23H28N4O2 Precise molecular weight: 392.22 LC-MS (m/z): 393.13 [M+H]+.
1HNMR (400 MHz, DMSO-d6) δ (ppm): 15.41 (s, 1H), 9.46 (d, J=6.0 Hz, 1H), 8.57 (s, 1H), 8.49 (d, J=6.4 Hz, 1H), 7.96 (s, 2H), 7.83 (s, 1H), 7.71 (d, J=6.8 Hz, 1H), 4.42 (s, 1H), 3.05 (d, J=8.0 Hz, 1H), 2.72 (d, J=10.0 Hz, 1H), 2.22 (s, 3H), 1.99-1.92 (m, 3H), 1.77-1.74 (m, 1H), 1.63-1.48 (m, 2H).
Molecular formula: C20H20N6O Precise molecular weight: 360.17 LC-MS (m/z): 361.09 [M+H]+.
1HNMR (400 MHz, DMSO-d6) δ (ppm): 10.22 (s, 1H), 8.58 (s, 1H), 8.40 (s, 1H), 8.21 (d, J=4.8 Hz, 1H), 7.92-7.88 (m, 1H), 7.85-7.81 (m, 1H), 7.67 (s, 1H), 7.47 (d, J=8.0 Hz, 1H), 7.47 (d, J=4.8 Hz, 1H), 4.75 (s, 1H), 3.56 (s, 2H), 3.23 (s, 1H), 2.74 (s, 4H), 2.01 (s, 2H), 1.83 (s, 2H).
Molecular formula: C19H21N5O Precise molecular weight: 335.17 LC-MS (m/z): 336.12 [M+H]+.
1HNMR (400 MHz, DMSO-d6) δ (ppm): 11.18 (d, J=5.2 Hz, 1H), 10.24 (s, 1H), 8.59 (s, 1H), 7.88-7.86 (m, 1H), 7.82-7.78 (m, 2H), 7.62 (s, 2H), 7.46 (d, J=7.6 Hz, 1H), 7.04-7.01 (m, 1H), 6.53 (d, J=7.2 Hz, 1H), 4.75 (s, 1H), 3.51 (s, 2H), 3.17 (s, 1H), 2.69 (s, 4H), 2.00 (s, 2H), 1.82 (s, 2H).
Molecular formula: C23H23N5O2 Precise molecular weight: 401.19 LC-MS (m/z): 402.13 [M+H]+.
1HNMR (400 MHz, DMSO-d6) δ (ppm): 10.48 (s, 1H), 8.39 (d, J=8.0 Hz, 1H), 7.88-7.84 (m, 1H), 7.81-7.77 (m, 1H), 7.50 (d, J=7.6 Hz, 1H), 7.43-7.41 (m, 1H), 7.33 (d, J=1.2 Hz, 1H), 7.19 (d, J=7.6 Hz, 1H), 4.56-4.52 (m, 2H), 4.49-4.44 (m, 3H), 3.48-3.45 (m, 1H), 3.04-3.01 (m, 1H), 2.67 (d, J=10.8 Hz, 1H), 2.05 (d, J=8.8 Hz, 1H), 1.88-1.77 (m, 3H), 1.67-1.61 (m, 1H), 1.55-1.49 (m, 1H).
Molecular formula: C23H23N5O2 Precise molecular weight: 401.19 LC-MS (m/z): 402.14 [M+H]+.
1H-NMR (300 MHz, DMSO-d6) δ (ppm): 11.92 (brs, 1H), 9.20-9.22 (m, 1H), 9.01 (s, 1H), 8.18-8.21 (d, 1H), 7.95-7.99 (m, 1H), 7.84 (s, 1H), 6.77 (s, 1H), 6.73 (s, 1H), 4.66 (s, 1H), 3.55 (s, 3H), 2.72 (s, 4H), 2.31 (s, 3H), 1.99-2.02 (d, 2H), 1.79-1.83 (d, 2H).
Molecular formula: C20H23N5O Precise molecular weight: 349.19 LC-MS (m/z): 350.22 [M+H]+.
1HNMR (400 MHz, DMSO-d6) δ (ppm): 9.31 (s, 1H), 8.31 (s, 1H), 7.74-7.72 (d, 1H), 7.60-7.58 (d, 1H), 7.31 (s, 1H), 7.06-7.03 (m, 1H), 6.73 (s, 2H), 4.68 (s, 1H), 2.57 (s, 4H), 2.50 (s, 3H), 2.31 (s, 3H), 2.06-1.99 (t, 5H), 1.83-1.81 (m, 2H).
The two-dimensional NOE spectrum showed coupling signals between 7.60-7.58 (d, 1H) and 7.74-7.72 (d, 1H) and coupling signals between 7.31 (s, 1H) and 8.31 (s, 1H).
Molecular formula: C21H26N4O Precise molecular weight: 362.21 LC-MS (m/z): 363.26 [M+H]+.
1HNMR (300 MHz, DMSO-d6) δ (ppm): 9.03 (s, 1H), 8.60 (s, 1H), 7.45-7.43 (d, 1H), 6.79-6.77 (d, 2H), 4.44 (s, 1H), 3.33 (s, 3H), 2.95 (s, 1H), 2.66 (s, 2H), 2.55 (s, 3H), 2.31 (s, 3H), 1.97-1.92 (d, 2H), 1.75-1.59 (m, 2H).
Molecular formula: C18H22N6O Precise molecular weight: 338.19 LC-MS (m/z): 339.27 [M+H]+.
1HNMR (300 MHz, DMSO-d6) δ (ppm): 9.66 (s, 1H), 9.11 (s, 1H), 8.29-8.26 (d, 1H), 7.72-7.70 (d, 1H), 7.20-7.17 (d, 1H), 6.85-6.79 (t, 2H), 4.73 (s, 1H), 3.38 (s, 5H), 2.80 (s, 3H), 2.55 (s, 2H), 2.34 (s, 3H), 2.08-2.02 (t, 2H), 1.88-1.84 (d, 2H).
Molecular formula: C22H26N4O3S Precise molecular weight: 426.17 LC-MS (m/z): 427.13 [M+H]+.
1HNMR (300 MHz, CDCl3) δ(ppm): 8.43 (s, 1H), 8.28-8.25 (d, 1H), 8.08-8.05 (d, 1H), 7.62-7.59 (d, 1H), 7.51-7.48 (d, 1H), 7.09-7.02 (d, 2H), 6.85-6.83 (d, 1H), 6.60 (s, 2H), 4.79 (s, 1H), 2.95-2.93 (d, 2H), 2.68 (s, 1H), 2.44-2.42 (d, 6H), 2.14 (s, 1H), 2.00 (s, 2H), 1.73 (s, 2H).
Molecular formula: C25H26N4O2 Precise molecular weight: 414.21 LC-MS (m/z): 415.21 [M+H]+.
1HNMR (300 MHz, DMSO-d6) δ (ppm): 7.51-7.48 (d, 1H), 7.37-7.34 (d, 2H), 7.23-7.21 (d, 1H), 6.85 (s, 1H), 6.80-6.78 (d, 1H), 4.57 (s, 1H), 3.50-3.42 (d, 2H), 2.44-3.42 (m, 2H), 3.41-3.40 (d, 1H), 3.16 (s, 6H), 3.06 (s, 1H), 2.57 (s, 3H), 2.34 (s, 3H), 2.02-1.99 (t, 2H), 1.78-1.74 (d, 2H).
Molecular formula: C23H29N5O Precise molecular weight: 391.24 LC-MS (m/z): 392.19 [M+H]+.
1H-NMR (400 MHz, DMSO-d6) δ (ppm): 10.48 (s, 1H), 6.73 (t, J=9.04 Hz, 1H), 6.61 (d, J=10.4 Hz, 1H), 5.77 (s, 1H), 4.33 (s, 1H), 3.07 (s, 1H), 2.77 (s, 1H), 2.37-2.35 (m, 7H), 1.84-1.51 (m, 10H).
Molecular formula: C20H24F2N4O Precise molecular weight: 374.19 LC-MS (m/z): 375.16 [M+H]+.
1H-NMR (400 MHz, DMSO-d6) δ (ppm): 9.69 (s, 1H), 7.24-7.20 (m, 1H), 7.12-7.09 (m, 1H), 6.92 (d, J=8.0 Hz, 1H), 6.80 (t, J=7.4 Hz, 1H), 5.77 (s, 1H), 4.41 (s, 1H), 3.21 (s, 1H), 3.04-2.92 (m, 2H), 2.48-2.33 (m, 8H), 1.87-1.58 (m, 8H).
Molecular formula: C20H26N4O Precise molecular weight: 338.21 LC-MS (m/z): 339.12 [M+H]+.
1H-NMR (400 MHz, DMSO-d6) δ (ppm): 9.60 (s, 1H), 6.86 (d, J=9.84 Hz, 1H), 6.77 (d, J=6.76 Hz, 1H), 5.87 (s, 1H), 4.42 (s, 1H), 3.31-3.20 (m, 2H), 3.16 (s, 1H), 2.99 (s, 1H), 2.40-2.35 (m, 5H), 2.20 (s, 3H), 1.88-1.59 (m, 10H).
Molecular formula: C21H27FN4O Precise molecular weight: 370.22 LC-MS (m/z): 371.21 [M+H]+.
1H-NMR (400 MHz, DMSO-d6) δ (ppm): 10.61 (s, 1H), 6.99-6.94 (m, 1H), 6.91-6.87 (m, 1H), 5.85 (s, 1H), 4.39 (s, 1H), 3.13 (s, 1H), 2.85 (s, 1H), 2.41-2.28 (m, 7H), 1.85-1.62 (m, 10H).
Molecular formula: C20H24F2N4O Precise molecular weight: 374.19 LC-MS (m/z): 375.22 [M+H]+.
1HNMR (300 MHz, DMSO-d6) δ (ppm): 8.40-8.37 (d, J=9 Hz, 1H), 7.87-7.82 (m, 2H), 7.58-7.55 (d, J=9 Hz, 1H), 7.05-7.02 (d, J=9 Hz, 1H), 6.96-6.93 (d, J=9 Hz, 1H), 6.65 (s, 1H), 6.53-6.50 (d, J=9 Hz, 1H), 4.89 (s, 2H), 4.43-4.41 (d, J=6 Hz, 1H), 3.10-3.08 (d, J=6 Hz, 1H), 2.73-2.70 (d, J=9 Hz, 1H), 2.27 (s, 3H), 2.22 (s, 3H), 1.99-1.40 (m, 6H).
Molecular formula: C21 H25N5 Precise molecular weight: 347.47 LC-MS (m/z): 348.25 [M+H]+.
1HNMR (300 MHz, DMSO-d6) δ (ppm): 8.45-8.42 (d, J=9 Hz, 1H), 7.88-7.75 (m, 2H), 7.51-7.49 (d, J=6 Hz, 1H), 7.24 (s, 1H), 6.96-6.93 (d, J=9 Hz, 1H), 6.56-6.53 (m, 2H), 5.03-5.02 (d, J=3 Hz, 1H), 4.56 (s, 1H), 2.98-2.95 (d, J=9 Hz, 1H), 2.63-2.62 (d, J=3 Hz, 3H), 2.46 (s, 3H), 2.34 (s, 3H), 2.28-1.60 (m, 6H).
Molecular formula: C22H27N5 Precise molecular weight: 361.49 LC-MS (m/z): 362.26 [M+H]+.
1HNMR (300 MHz, DMSO-d6) δ (ppm): 9.63 (s, 1H), 8.48 (s, 1H), 7.89-7.77 (m, 2H), 7.54-7.52 (m, 1H), 7.41 (s, 1H), 7.17-7.15 (m, 1H), 6.82-6.76 (m, 2H), 4.65 (s, 1H), 3.47-3.36 (m, 2H), 3.17-3.12 (m, 2H), 2.51-2.50 (m, 3H), 2.33 (s, 3H), 1.99-1.77 (m, 4H).
Molecular formula: C22H26N4O Precise molecular weight: 362.48 LC-MS (m/z): 363.33 [M+H]+.
1HNMR (300 MHz, DMSO-d6) δ (ppm): 9.19 (s, 1H), 8.41-8.38 (m, 1H), 7.86-7.64 (m, 3H), 7.39-7.36 (m, 1H), 7.29-7.26 (m, 1H), 7.12-7.10 (m, 2H), 4.45-4.43 (m, 1H), 3.14-3.11 (m, 1H), 2.79-2.77 (m, 1H), 2.39 (s, 3H), 2.25 (s, 3H), 2.01-1.91 (m, 3H), 1.80-1.54 (m, 6H).
Molecular formula: C23H27N5O Precise molecular weight: 389.22 LC-MS (m/z): 390.30 [M+H]+.
1HNMR (300 MHz, DMSO-d6) δ (ppm): 8.51 (s, 1H), 7.86-7.68 (m, 3H), 7.53 (s, 1H), 7.46-7.06 (m, 4H), 4.63 (s, 1H), 3.02 (s, 1H), 2.56 (s, 3H), 2.37 (s, 3H), 2.04-1.34 (m, 7H).
Molecular formula: C22H25N5O Precise molecular weight: 375.21 LC-MS (m/z): 376.22 [M+H]+.
1HNMR (300 MHz, DMSO-d6) δ (ppm): 9.61 (s, 1H), 8.40-8.38 (m, 1H), 7.88-7.76 (m, 2H), 7.54-7.52 (m, 1H), 7.19-7.17 (m, 2H), 6.81-6.76 (m, 2H), 6.14-6.05 (m, 1H), 5.70-5.59 (m, 1H), 4.70-4.00 (m, 3H), 3.35-2.71 (m, 2H), 2.33 (s, 3H), 2.15-1.24 (m, 5H).
Molecular formula: C23H24N4O2 Precise molecular weight: 388.19 LC-MS (m/z): 389.12 [M+H]+.
1HNMR (300 MHz, DMSO-d6) δ (ppm): 9.99 (s, 1H), 8.51-8.47 (m, 2H), 7.90-7.78 (m, 2H), 7.66 (s, 1H), 7.49-7.34 (m, 4H), 4.76 (s, 1H), 3.56 (s, 1H), 3.24-3.09 (m, 2H), 2.89-2.75 (m, 6H), 2.13-1.80 (m, 5H).
Molecular formula: C22H25N5O2 Precise molecular weight: 391.20 LC-MS (m/z): 392.14 [M+H]+.
1HNMR (400 MHz, DMSO-d6) δ (ppm): 9.96 (s, 1H), 8.47 (s, 1H), 7.89-7.79 (m, 2H), 7.67-7.65 (d, J=8 Hz, 1H), 7.37 (s, 1H), 7.21-7.19 (d, J=8 Hz, 1H), 6.59-6.53 (m, 2H), 4.63 (s, 1H), 3.49 (s, 3H), 3.16-3.14 (m, 5H), 2.65 (s, 3H), 2.03-1.77 (m, 4H).
Molecular formula: C25H29N5O2 Precise molecular weight: 419.23 LC-MS (m/z): 420.19 [M+H]+.
1HNMR (400 MHz, DMSO-d6) δ (ppm): 10.53 (s, 1H), 7.35-7.24 (m, 3H), 5.69-5.68 (m, 1H), 4.29 (s, 1H), 3.09 (s, 1H), 2.83 (s, 1H), 2.37-2.09 (m, 9H), 1.86-1.70 (m, 4H), 1.61-1.46 (m, 4H), 0.87-0.84 (m, 2H), 0.47-0.44 (m, 2H).
Molecular formula: C24H29N5O Precise molecular weight: 403.24 LC-MS (m/z): 404.15 [M+H]+.
1HNMR (400 MHz, DMSO-d6) δ (ppm): 10.50 (s, 1H), 7.33 (s, 2H), 7.24 (s, 1H), 5.89-5.77 (m, 1H), 4.49-4.10 (m, 3H), 3.04-2.90 (m, 1H), 2.67-2.59 (m, 1H), 2.41-2.28 (m, 4H), 2.02-2.01 (m, 4H), 1.76-1.60 (m, 7H).
Molecular formula: C22H25N5O2 Precise molecular weight: 391.20 LC-MS (m/z): 392.11 [M+H]+.
1HNMR (400 MHz, DMSO-d6) δ (ppm): 11.06 (s, 1H), 7.80-7.71 (m, 3H), 7.59-7.57 (m, 1H), 7.30-7.28 (m, 1H), 6.94-6.81 (m, 2H), 4.70 (s, 1H), 3.68-6.65 (m, 1H), 3.17 (s, 3H), 2.68 (s, 3H), 2.42-2.39 (m, 2H), 2.16-2.12 (m, 2H), 1.60-1.53 (m, 2H).
Molecular formula: C21H24N4O3S Precise molecular weight: 412.16 LC-MS (m/z): 413.09 [M+H]+.
1HNMR (400 MHz, DMSO-d6) δ (ppm): 10.47 (s, 1H), 8.42-8.40 (d, J=8 Hz, 1H), 7.89 (t, J=8 Hz, 1H), 7.81 (t, J=8 Hz, 1H), 7.52-7.50 (d, J=8 Hz, 1H), 7.44-7.42 (d, J=8 Hz, 2H), 7.34-7.33 (m, 2H), 4.42 (s, 1H), 4.03-4.00 (m, 1H), 3.63-3.60 (m, 1H), 2.91-2.86 (m, 1H), 2.78-2.73 (m, 1H), 2.68-2.62 (m, 1H), 2.08-1.92 (m, 2H), 1.69-1.68 (m, 2H) 1.01-0.92 (m, 4H).
Molecular formula: C23H23N5O3S Precise molecular weight: 449.15 LC-MS (m/z): 450.07 [M+H]+.
1HNMR (400 MHz, DMSO-d6) δ (ppm): 10.46 (s, 1H), 8.43-8.41 (d, J=8 Hz, 1H), 7.89 (t, J=8 Hz, 1H), 7.82-7.81 (m, 1H), 7.52-7.50 (d, J=8 Hz, 1H), 7.43-7.42 (d, J=4 Hz, 1H), 7.33-7.32 (m, 2H), 4.43 (s, 1H), 4.01-3.98 (m, 1H), 3.56-3.53 (m, 1H), 2.90 (s, 3H), 2.80-2.75 (m, 1H), 2.65-2.60 (m, 1H), 2.06-1.93 (m, 2H), 1.71-1.69 (m, 2H).
Molecular formula: C21H21N5O3S Precise molecular weight: 423.14 LC-MS (m/z): 424.06 [M+H]+.
1HNMR (400 MHz, DMSO-d6) δ (ppm): 11.17 (s, 1H), 10.76 (s, 1H), 8.03-7.97 (m, 3H), 7.64-7.57 (m, 2H), 7.49-7.44 (m, 2H), 3.55 (s, 1H), 3.22 (s, 2H), 3.11-3.09 (m, 1H), 2.89 (s, 1H), 2.75 (s, 3H), 2.24 (s, 1H), 2.01-1.65 (m, 4H).
Molecular formula: C22H21N5O2 Precise molecular weight: 387.17 LC-MS (m/z): 388.09 [M+H]+.
1HNMR (400 MHz, DMSO-d6) δ (ppm): 11.07 (s, 1H), 10.15 (s, 1H), 9.00-8.98 (d, J=8 Hz, 1H), 8.16-8.06 (m, 2H), 7.61-7.55 (m, 2H), 7.50-7.47 (m, 2H), 5.37 (s, 1H), 4.09-3.90 (m, 2H), 2.90-2.88 (m, 2H), 2.27-2.20 (m, 2H).
Molecular formula: C19H16N4O2 Precise molecular weight: 332.13 LC-MS (m/z): 333.02 [M+H]+.
1HNMR (400 MHz, DMSO-d6) δ (ppm): 11.06 (s, 1H), 10.33 (s, 1H), 7.81-7.73 (m, 3H), 7.54-7.52 (d, J=8 Hz, 1H), 7.25-7.23 (d, J=8 Hz, 1H), 6.87-6.83 (m, 2H), 4.70 (s, 1H), 3.69-3.66 (m, 1H), 3.46-3.40 (m, 1H), 3.14 (s, 3H), 2.43-2.33 (m, 1H), 2.13-1.97 (m, 1H), 1.61-1.55 (m, 2H).
Molecular formula: C21H24N4O2S Precise molecular weight: 396.16 LC-MS (m/z): 397.08 [M+H]+.
1HNMR (400 MHz, DMSO-d6) δ (ppm): 8.34-8.32 (d, J=8 Hz, 1H), 7.92-7.88 (m, 1H), 7.83-7.79 (m, 1H), 7.50-7.39 (m, 3H), 7.34-7.30 (m, 2H), 4.71-4.61 (m, 1H), 3.18-3.03 (m, 2H), 2.49 (s, 1H), 2.51 (s, 1H), 2.40-2.29 (m, 4H), 2.10-2.05 (m, 2H).
Molecular formula: C21H19F2N5O2 Precise molecular weight: 395.16 LC-MS (m/z): 396.07 [M+H]+.
1HNMR (400 MHz, DMSO-d6) δ (ppm): 10.47 (s, 1H), 8.36-8.34 (d, J=8 Hz, 1H), 7.89 (t, J=8 Hz, 1H), 7.80 (t, J=8 Hz, 1H), 7.50-7.49 (m, 1H), 7.43-7.41 (m, 2H), 7.33-7.28 (m, 2H), 4.86-4.48 (m, 2H), 3.09-3.07 (m, 2H), 2.44-2.43 (m, 1H), 2.29 (s, 3H), 2.06-1.90 (m, 2H), 1.77-1.66 (m, 2H).
Molecular formula: C21H20FN5O Precise molecular weight: 377.17 LC-MS (m/z): 378.08 [M+H]+.
1HNMR (400 MHz, DMSO-d6) δ (ppm): 8.39-8.37 (d, J=8 Hz, 1H), 7.87 (t, J=8 Hz, 1H), 7.79 (t, J=8 Hz, 1H), 7.50-7.48 (d, J=8 Hz, 1H), 7.43-7.41 (m, 2H), 7.34-7.28 (m, 2H), 4.95 (s, 1H), 4.47-4.46 (m, 1H), 3.72-3.67 (m, 1H), 3.17-3.09 (m, 1H), 2.90-2.89 (m, 1H), 2.20-2.27 (m, 4H), 1.91-1.80 (m, 2H), 1.47-1.34 (m, 2H).
Molecular formula: C21H21N5O2 Precise molecular weight: 375.17 LC-MS (m/z): 376.10 [M+H]+.
1HNMR (400 MHz, DMSO-d6) δ (ppm): 13.47 (s, 1H), 7.71-7.69 (d, J=8.1 Hz, 1H), 7.35-7.32 (m, 2H), 6.62 (s, 1H), 4.37 (s, 1H), 3.30 (s, 1H), 3.19-3.17 (m, 1H), 3.11-3.07 (m, 2H), 2.90 (s, 1H), 2.82-2.79 (m, 2H), 2.43 (s, 3H), 2.25 (br, 1H), 2.11-2.03 (m, 2H), 1.91 (m, 1H), 1.81 (s, 1H), 1.68-1.64 (m, 1H), 1.50 (s, 1H).
Molecular formula: C20H23N5O Precise molecular weight: 349.19 LC-MS (m/z): 350.09 [M+H]+.
1HNMR (400 MHz, DMSO-d6) δ (ppm): 12.91 (s, 1H), 7.70-7.68 (d, J=8.4 Hz, 1H), 7.38-7.36 (m, 2H), 6.96 (s, 1H), 4.44 (s, 1H), 4.11 (s, 1H), 3.78 (s, 1H), 3.60 (s, 1H), 3.17-3.10 (m, 3H), 2.62 (s, 4H), 2.02-1.91 (m, 4H), 1.74 (s, 1H), 1.64-1.60 (m, 1H), 1.52-1.50 (m, 1H), 1.23-1.20 (m, 1H), 1.10-1.00 (m, 1H).
Molecular formula: C22H25N5O Precise molecular weight: 375.21 LC-MS (m/z): 376.22 [M+H]+.
Test Samples: The compounds of the present invention prepared according to exemplary methods.
The test results are shown in Table 2 below.
As can be seen from the experimental results in Table 2, the compounds of the present invention have good inhibitory activities against NLRP3 inflammasomes, and thus the compounds of the present invention can be used for preventing and/or treating NLRP3 inflammasome-associated diseases.
The above description is only for the purpose of illustrating preferred examples of the present invention, and is not intended to limit the scope of the present invention. Any modifications, equivalents, improvements, and the like made without departing from the spirit and principle of the present invention should be included in the protection scope of the present invention.
| Number | Date | Country | Kind |
|---|---|---|---|
| 202110629439.0 | Jun 2021 | CN | national |
| 202110843496.9 | Jul 2021 | CN | national |
| 202110940013.7 | Aug 2021 | CN | national |
| 202110940018.X | Aug 2021 | CN | national |
| 202210015181.X | Jan 2022 | CN | national |
| 202210015699.3 | Jan 2022 | CN | national |
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/CN2022/096928 | 6/2/2022 | WO |
