Patent Application
20250179087
The present invention relates to novel compounds; methods for the production of the compounds of the invention; pharmaceutical compositions comprising the compounds of the invention; as well as uses and methods for treating a disease mediated by LPAR5 by administering the compounds of the invention. In particular, the compounds of the invention may be used as LPAR5 antagonists.
Lysophosphatidic acid (LPA) is a small ubiquitous lipid found in vertebrate and nonvertebrate organisms that mediate diverse biological actions and demonstrate medicinal relevance. LPA is a soluble, bioactive signaling lipid derived largely from membrane phospholipids. It is normally present at biologically relevant concentrations in blood plasma and is further enriched by release from activated platelets under inflammatory conditions. LPA-producing lysoPLD (Autotaxin) converts lysophosphatidyl choline (LPC) to LPA, and LPA binds to each LPA receptor. The distribution of LPA receptors in different cell types results in differential signals and modulates cellular functions. LPA signaling is involved in mediating a wide range of cellular processes and has been linked to numerous disease states, including neurological disorders, inflammation, and cancer.
LPA exerts its cellular functions by acting on specific G protein-coupled receptors (GPCR). There are at least nine GPCRs that were reported to be activated by LPA, amongst which LPAR1, LPAR2, and LPAR3 receptors have been extensively studied. LPAR1-3 receptors belong to the same endothelial differentiation gene (EDG) GPCR subfamily as the sphingosine-1-phosphate receptors. Little is known about the roles of LPAR4 (GPR23/P2Y9) and LPAR6 (P2Y5) receptors in cancer and characterization of the putative LPA GPCR GPR87, P2Y10, and GPR35 awaits further research.
LPAR5 (also named as LPA5) was first discovered as an orphan GPCR, GPR92/GPR93. LPAR5 shares 35% homology with LPAR4, but only ˜22% homology to LPARI-3. LPAR5/Lpar5 (human chromosome 12pl3.31; mouse chromosome 6, 59.21 cM) encodes a 372 amino acid protein. LPAR5 couples to G12/13 and Gq/11 to increase intracellular cyclic adenosine monophosphate (cAMP) levels. LPAR5 is highly expressed in the spleen, and to a lesser degree in the heart, small intestine, placenta, colon, and liver. It is highly expressed in the spinal cord and dorsal root ganglion (DRG), nervous system structures that are associated with pain.
Abound evidence indicates that LPAR5 is involved in broad pain signaling in the spinal cord. LPAR5 is upregulated in the dorsal horn of the spinal cord in a rat model of neuropathic pain. LPAR5-deficient mice do not develop mechanical allodynia in a model of neuropathic pain and develop a lesser degree of cold allodynia induced by nerve injury compared with wild-type mice. In addition, LPAR5-deficient mice show decreased pain sensitivity in tail withdrawal tests and accelerated recovery in thermal hyperalgesia. Injection of LPAR5 agonists induces pain. A novel LPA5 antagonist with high potency, selectivity, and central nervous system penetrability, with broader analgesic effects than pregabalin and duloxetine in multiple animal models of pain sensitizers-induced allodynia, neuropathic pain, and inflammatory pain. LPAR5 is involved in broad pain signaling in the spinal cord and that the pharmacological antagonism of LPAR5 is an attractive novel pain therapy.
LPA was well studied as a growth factor and mitogen for tumor cell migration, invasion, and angiogenesis. LPAR5 was also recently demonstrated in involvement in tumor progression. It has recently been shown that the LPAR5 receptor may potentially inhibit the migration of B16 melanoma cells by activating the cAMP-PKA pathway and diminishing PIP3 signaling. LPA engagement with the LPAR5 receptor induces a signal that inhibits TCR-induced Ca2+ release from intracellular stores in naïve CD8 T cells. Cancer cells produce a large amount of LPA, the abundant evidence indicates that an LPA-LPAR5 signaling axis is also exploited by diverse cancers to suppress T cell activation and function.
In addition to cancer and neuropathic pain, deregulated LPA-LPAR5 signaling may be also involved in other diseases such as neurological disorders, atherosclerosis and cardiovascular disease, inflammatory and autoimmune diseases, fibrosis, bone development and disease, reproductive system and infertility, or obesity.
The important role of LPAR5 in multiple diseases including neurodegeneration, neuroinflammation, cancer development and autoimmune diseases suggests that targeted inhibition of LPAR5 is a potential therapeutic approach for the treatment of neurodegenerative diseases, cancer, autoimmune diseases, and other inflammatory diseases. One of the most important strategies targeting LPAR5 is to develop small molecule inhibitors against LPAR5, which have demonstrated promising preclinical efficacy.
Provided are a series of novel compounds as LPAR5 inhibitors. The inventors of the instant invention have found that the compounds disclosed herein show better solubility and excellent pharmacokinetics (such as higher kinetic solubility and longer hLM t1/2 and mLM t1/2 reflecting the stability of the compounds) by tuning the moiety Cyc and/or replacing X4 with a nitrogen atom.
Provided herein is a compound of formula (1)
or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof or a deuterated derivative thereof, wherein
In some embodiments, X1 is N, X2, X3 and X4 are CR2, CR3 and CR4, respectively; or X2 is N, X1, X3 and X4 are CR1, CR3 and CR4, respectively; or X3 is N, X1, X2 and X4 are CR1, CR2 and CR4, respectively; or X4 is N, X2, X3 and X4 are CR2, CR3 and CR4, respectively; or X1 is N, X4 is N, X2 is CR2, and X3 is CR3; or X1 is CR1, X4 is CR4, X2 is N, and X3 is N; wherein R1, R2, R3, R4, at each of their occurrences, is independent as defined above. In some embodiments, X2 is CR2, and X3 is CR3, and X1 is CR1 or N, and X4 is CR4 or N.
In some embodiments, is a double bond and X7 is N; or
is a double bond and X7 is CR7a.
In some embodiments, the compound of Formula (I) is
wherein each of the variables is defined as in Formula (I).
In some embodiments, R1, R2, R3 and R4 are each independently hydrogen, halogen, C1-6alkyl, C1-6alkoxy, C3-8cycloalkyl, C3-8cycloalkoxy or heterocyclyloxy, each of said C1-6alkyl, C1-6alkoxy, C3-8cycloalkyl, C3-8cycloalkoxy or heterocyclyloxy is unsubstituted or substituted with one or two or three halogen, deuterium or C1-4alkoxy, provided that R2 and R3 are not both hydrogen.
In some embodiments, R2 and R3 are each independently hydrogen, halogen, C1-6alkyl, C1-6alkoxy, C3-8cycloalkyl, C3-8cycloalkoxy, or heterocyclyloxy, each of said C1-6alkyl, C1-6alkoxy, C3-8cycloalkyl, C3-8cycloalkoxy and heterocyclyloxy is unsubstituted or substituted with one or two or three halogen, deuterium or C1-4alkoxy. In some embodiments, R2 and R3 are each independently halogen, C1-4 alkyl, C1-4alkoxy, C3-6cycloalkyl, or C3-8cycloalkoxy, each of said C1-4alkyl, C1-4alkoxy, C3-6cycloalkyl, and C3-6cycloalkoxy is unsubstituted or substituted with one or two or three halogen or deuterium. In some further embodiments, R2 and R3 are each independently methoxy, fluoro, methoxy-d3, cyclopropoxy, fluoromethoxy, difluoromethoxy, trifluoromethoxy, azetidin-3-yloxy, oxetan-3-yloxy, cyclopropylmethoxy, 2-methoxyethoxy. In some further embodiments, R2 and R3 are each independently hydrogen, methoxy, methoxy-d3, provided that R2 and R3 are not both hydrogen. In some embodiments, R2 is hydrogen and R3 is methoxy or methoxy-d3. In some further embodiments, R2 is methoxy or methoxy-d3 and R3 is methoxy or methoxy-d3. In some further embodiments, R2 is methoxy and R3 is methoxy-d3; or R2 is methoxy-d3 and R3 is methoxy; or R2 and R3 are each methoxy; or R2 and R3 are each methoxy-d3.
In some embodiments, R2 and R3 together with the atom to which they are attached, form a 4-, 5-, 6-, or 7-membered heterocyclyl ring comprising 0, 1 or 2 heteroatom(s) independently selected from nitrogen, oxygen or optionally oxidized sulfur as ring member(s), said ring is unsubstituted or substituted with at least one substituent R, wherein R is halogen, C1-6alkyl, C3-8cycloalkyl, —ORa, —NRbRc, phenyl, heteroaryl, or heterocyclyl, wherein each of Ra, Rb, and Rc is hydrogen, C1-4alkyl, phenyl, heteroaryl, or heterocyclyl.
In some embodiments, R2 and R3 together with the atom to which they are attached, form the following structure
wherein R is hydrogen, halogen, C1-6alkyl or C1-6alkoxy, each of said C1-6alkyl and C1-6alkoxy is unsubstituted or substituted with one or two or three halogen or deuterium, n is 0, 1 or 2; and the other variables are defined as above. In some further embodiments, n is 2, and R is halogen, preferably fluoro; or C1-4alkyl, preferably methyl.
In some embodiments, R5 is
In some embodiments, R5 is —C(O)NR5aR5b, wherein R5a is hydrogen, C1-6alkyl, C3-8cycloalkyl, or C3-6cycloalkylC1-6alkyl-; preferably methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, pentyl, hexyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclopropylmethyl, cyclobutylmethyl, cyclopentylmethyl, or cyclohexylmethyl; and R5b is hydrogen or C1-6alkyl. In some further embodiments, R5 is N,N-dimethylaminocarbonyl, N-cyclopropyl-N-methylaminocarbonyl, N-cyclopropylaminocarbonyl, N-cyclopropyl-N-(cyclopropylmethyl)aminocarbonyl, or N-cyclobutyl-N-methylaminocarbonyl.
In some embodiments, R5 is —C(O)NR5aR5b, wherein R5a and R5b, together with the nitrogen atom to which they are attached, form a 4-, 5-, 6-, or 7-membered heterocyclyl ring comprising 0 or 1 additional heteroatom independently selected from nitrogen, oxygen or optionally oxidized sulfur as ring member, said ring is unsubstituted or substituted with one or two substituents R, wherein R is halogen, deuterium, C1-4alkyl, deuterated C1-4alkyl or —ORa, wherein Ra is hydrogen or C1-4alkyl or deuterated C1-4 alkyl; preferably fluoro, bromo, chloro, methyl, methyl-d3, ethyl, propyl, isopropyl, butyl, isobutyl, or tertbutyl. In some further embodiments, R5 is —C(O)NR5aR5b, wherein R5a and R5b, together with the nitrogen atom to which they are attached, form a 4-, 5-, 6-, or 7-membered heterocyclyl ring, which is azetidine-1-yl, pyrrolidine-1-yl, piperidine-1-yl, piperazine-1-yl, morpholine-4-yl, each of said ring is unsubstituted or substituted with one or two fluoro, bromo, chloro, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, or tertbutyl; preferably fluoro, or methyl. In some further embodiments, R5 is —C(O)NR5aR5b, wherein R5a and R5b, together with the nitrogen atom to which they are attached, form a 5- or 6-membered heterocyclyl ring, which is pyrrolidine-1-yl, or piperidine-1-yl, each of said ring is unsubstituted or substituted with fluoro.
In some even further embodiments, R5 is 3,3-difluoroazetidine-1-carbonyl, 3,3-difluoropyrrolidine-1-carbonyl, piperidine-1-carbonyl, piperazine-1-carbonyl, 4-methylpiperazine-1-carbonyl, morpholine-4-carbonyl, pyrrolidine-1-carbonyl, 3-hydroxypyrrolidine-1-carbonyl, 4-fluoropiperidine-1-carbonyl, 4,4-difluoropiperidine-1-carbonyl, or morpholine-4-carbonyl.
In some embodiments, R5 is —C(O)NR5aR5b, wherein R5a and R5b, together with the nitrogen atom to which they are attached, form a 4-, 5-, 6-, or 7-membered heterocyclyl ring comprising 0 or 1 additional heteroatom independently selected from nitrogen, oxygen or optionally oxidized sulfur as ring member, said ring is unsubstituted or substituted with one or two substituents R; wherein two R attached to the same carbon atom in the ring form a spiro C3-C6 carbon ring, preferably a spiro C3 carbon ring, said spiro C3-C6 carbon ring comprises 0 or 1 additional heteroatom independently selected from nitrogen, oxygen or optionally oxidized sulfur as ring member(s), and said spiro C3-C6 carbon ring is unsubstituted or substituted with one or two substituents selected from halogen or C1-4alkyl. In further embodiments, R5 is 6-azaspiro[2.5]octane-6-yl, 5-azaspiro[2.5]octane-5-yl, or 4-oxa-7-azaspiro[2.5]octane-7-yl.
In some embodiments, R5 is —C(O)NR5aR5b, wherein R5a and R5b, together with the nitrogen atom to which they are attached, form a 4-, 5-, 6-, or 7-membered heterocyclyl ring comprising 0 or 1 additional heteroatom independently selected from nitrogen, oxygen or optionally oxidized sulfur as ring member, said ring is unsubstituted or substituted with one or two substituents R; wherein two R attached to the two adjacent carbon atoms in the ring form a fused C3-C5 carbon ring; said fused C3-C6 carbon ring comprises 0 or 1 additional heteroatom independently selected from nitrogen, oxygen or optionally oxidized sulfur as ring member, and said fused C3-C6 carbon ring is unsubstituted or substituted with one or two substituents selected from halogen or C1-4alkyl. In some further embodiments, R5 is 6,6-dimethyl-3-azabicyclo[3.1.0]hexane-3-yl.
In some embodiments, R5 is piperidine-1-carbonyl, piperazine-1-carbonyl, 4-methylpiperazine-1-carbonyl, morpholine-4-carbonyl, pyrrolidine-1-carbonyl, 3-hydroxypyrrolidine-1-carbonyl, 4-fluoropiperidine-1-carbonyl, 4,4-difluoropiperidine-1-carbonyl, morpholine-4-carbonyl, 6-azaspiro[2.5]octane-6-carbonyl, 5-azaspiro[2.5]octane-5-carbonyl, (4-oxa-7-azaspiro[2.5]octane-7-carbonyl, 6,6-dimethyl-3-azabicyclo[3.1.0]hexane-3-carbonyl, 3,3-difluoropyrrolidine-1-carbonyl, 3,3-difluoroazetidine-1-carbonyl; or 2-azabicyclo[2.2.1]heptane-2-carbonyl.
In some embodiments, R5 is —C(O)R5a, wherein R5a is phenyl, which is unsubstituted or substituted with one or two halogen or C1-4alkyl. In some further embodiments, R5 is phenylcarbonyl, or 2-chlorophenylcarbonyl.
In some embodiments, R5 is —C(OH)R5aR5b, wherein R5a and R5b are each independently hydrogen, C1-6alkyl, or phenyl, each of said C1-6alkyl, or phenyl is unsubstituted or substituted with one or two halogen. In some further embodiments, R5 is hydroxy(methoxy(methyl)amino)(phenyl)methyl, hydroxymethyl, hydroxy(phenyl)methyl, or 2-chlorophenyl)(hydroxy)methyl.
In some embodiments, Cyc is phenyl, thiazolyl, pyridinyl, pyrazolyl, oxadiazolyl, isoxazolyl, tetrahydronaphthalenyl, dihydrondenyl, benzoisothiazolyl, benzoisoxazolyl, tetrahydroquinolinyl, dihydroquinolinyl, tetrahydroisoquinolinyl, isoquinolinyl, indazolyl, indolyl (such as indol-1-yl, indol-2-yl, indol-3-yl, indol-4-yl, indol-5-yl, indol-6-yl or indol-7-yl), isoindolyl (such as isoindol-1-yl, isoindol-2-yl, isoindol-3-yl, isoindol-4-yl, isoindol-5-yl, isoindol-6-yl or isoindol-7-yl), indolinyl, dihydrobenzofuranyl, benzofuranyl, or pyrrolopyridinyl, each of which is unsubstituted or substituted with one or two or three substituents selected from halogen, deuterium, C1-6alkyl, cyano, oxo, —ORd, —C(O)Rd, —C(O)NRdRe, —NRdRe, C3-8cycloalkyl, phenyl, heteroaryl, and heterocyclyl, each of said C1-6alkyl, C3-8cycloalkyl, phenyl, heteroaryl, and heterocyclyl is unsubstituted or substituted with halogen, deuterium, hydroxy, C1-6alkyl, C1-6alkoxy, or C3-8cycloalkyl, and wherein Rd and Rc are each hydrogen, deuterated C1-6alkyl, C1-6alkyl, C3-8cycloalkyl, phenyl, heteroaryl, or heterocyclyl.
In some embodiments, Cyc is unsubstituted or substituted with one or two or three substituents selected from halogen, deuterium, cyano, C1-6alkyl, oxo, —ORd, —C(O)Rd, C3-8cycloalkyl, or heterocyclyl, said C1-6alkyl is unsubstituted or substituted with halogen, deuterium, hydroxy, or C3-6cycloalkyl, wherein Rd is hydrogen, C1-4alkyl, deuterated C1-4alkyl or C3-8cycloalkyl. In some embodiments, Cyc is unsubstituted or substituted with one or two or three substituents selected from fluoro, chloro, methyl, ethyl, isopropyl, n-propyl, methyl-d3, oxo, trifluoromethyl, oxetan-3-yl, cyclopropylmethyl, cyclopropyl, acetyl, methoxy, methoxy-d3 or 2-hydroxyethyl.
In some embodiments, Cyc is thiazol-2-yl, pyrazol-3-yl, oxadiazol-3-yl, isoxazol-3-yl, pyridin-2-yl, pyridin-4-yl, pyridin-3-yl, 5,6,7,8-tetrahydronaphthalen-2-yl, 5,6,7,8-tetrahydronaphthalen-1-yl, 1,2,3,4-tetrahydronaphthalen-2-yl, 1,2,3,4-tetrahydronaphthalen-1-yl, 2,3-dihydro-1H-inden-1-yl, 2,3-dihydro-1H-inden-4-yl, benzo[d]isothiazol-3-yl, benzo[d]isoxazol-3-yl, 1,2,3,4-tetrahydroquinolin-7-yl, 1,2,3,4-tetrahydroquinolin-5-yl, 5,6,7,8-tetrahydroquinolin-1-yl, 5,6,7,8-tetrahydroquinolin-3-yl, 3,4-dihydroquinolin-7-yl, 5,6,7,8-tetrahydro-1H-isoquinolin-7-yl, 1,2,3,4-tetrahydro-1H-isoquinolin-8-yl, 1,2,3,4-tetrahydro-1H-isoquinolin-5-yl, isoquinolin-2-yl, 1H-indazol-4-yl, 1H-indazol-3-yl, 1H-indol-4-yl, 1H-indol-3-yl, 6-fluoro-1H-indol-4-yl, 1H-indol-6-yl, indolin-4-yl, 2,3-dihydrobenzofuran-4-yl, 1H-pyrrolo[3,2-c]pyridin-4-yl, 1H-pyrrolo[3,2-c]pyridin-3-yl or 1H-pyrrolo[2,3-b]pyridin-4-yl, each of which is unsubstituted or substituted as discussed above.
In some embodiments, Cyc is indol-4-yl, wherein position 6 of the indolyl ring is substituted with halogen (preferably fluoro) and position 1 of the indolyl ring is unsubstituted or substituted with C1-6alkyl (preferably methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tertbutyl), deuterated C1-6alkyl, C3-8cycloalkyl (preferably cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl), or hydoxyC1-6alkyl (preferably hydoxymethyl, hydoxyethyl, hydoxypropyl, or hydoxybutyl), and the other position is unsubstituted or substituted with halogen, C1-6alkyl or deuterated C1-6alkyl. In some embodiments, Cyc is indol-4-yl, wherein position 6 of the indolyl ring is unsubstituted and position 1 of the indolyl ring is substituted with C3-8cycloalkyl (preferably cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl), hydoxyC1-6alkyl (preferably hydoxymethyl, hydoxyethyl, hydoxypropyl, or hydoxybutyl) or —C(O)Rd, wherein Rd is hydrogen, C1-4alkyl, or C3-8cycloalkyl. In some embodiments, Cyc is indol-5-yl, wherein position 4 of the indolyl ring is substituted with halogen (preferably fluoro) and position 2 of the indolyl ring is unsubstituted or substituted with C1-6alkyl (preferably methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tertbutyl), deuterated C1-6alkyl, C3-8cycloalkyl (preferably cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl), or hydoxyC1-6alkyl (preferably hydoxymethyl, hydoxyethyl, hydoxypropyl, or hydoxybutyl), and the other position is unsubstituted or substituted with halogen, C1-6alkyl or deuterated C1-6alkyl. In some embodiments, Cyc is indol-5-yl, wherein position 4 of the indolyl ring is unsubstituted and position 1 of the indolyl ring is substituted with C3-8cycloalkyl (preferably cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl), hydoxyC1-6alkyl (preferably hydoxymethyl, hydoxyethyl, hydoxypropyl, or hydoxybutyl) or —C(O)Rd, wherein Rd is hydrogen, C1-4alkyl, or C3-8cycloalkyl. In some embodiments, Cyc is indol-6-yl, wherein position 4 of the indolyl ring is substituted with halogen (preferably fluoro) and position 1 of the indolyl ring is unsubstituted or substituted with C1-6alkyl (preferably methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tertbutyl), deuterated C1-6alkyl, C3-8cycloalkyl (preferably cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl), or hydoxyC1-6alkyl (preferably hydoxymethyl, hydoxyethyl, hydoxypropyl, or hydoxybutyl), and the other position is unsubstituted or substituted with halogen, C1-6alkyl or deuterated C1-6alkyl. In some embodiments, Cyc is indol-6-yl, wherein position 4 of the indolyl ring is unsubstituted and position 1 of the indolyl ring is substituted with C3-8cycloalkyl (preferably cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl), hydoxyC1-6alkyl (preferably hydoxymethyl, hydoxyethyl, hydoxypropyl, or hydoxybutyl) or —C(O)Rd, wherein Rd is hydrogen, C1-4alkyl, or C3-6cycloalkyl. In some embodiments, Cyc is indol-4-yl, indol-5-yl or indol-6-yl, wherein position 1 of the indolyl ring is substituted with C1-6alkyl (preferably methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tertbutyl), deuterated C1-6alkyl, C3-8cycloalkyl (preferably cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl), or hydoxyC1-6alkyl (preferably hydoxymethyl, hydoxyethyl, hydoxypropyl, or hydoxybutyl).
In some embodiments, Cyc is thiazol-2-yl, 4-methylpyridin-2-yl, 1-methyl-1H-pyrazol-3-yl, 5-isopropyl-1,2,4-oxadiazol-3-yl, 5-phenylisoxazol-3-yl, 2-cyanophenyl, 3-methylpyridin-2-yl, 4-fluoropyridin-2-yl, 4-cyanopyridin-2-yl, 2-fluoropyridin-4-yl, 3-cyanopyridin-2-yl, 2-(methylamino)pyridin-4-yl, 6-(methylamino)pyridin-3-yl, 3-fluoro-5-(pyrrolidin-1-yl)phenyl, 5,6,7,8-tetrahydronaphthalen-2-yl, 5,6,7,8-tetrahydronaphthalen-1-yl, 1,2,3,4-tetrahydronaphthalen-2-yl, 1,2,3,4-tetrahydronaphthalen-1-yl, 2,3-dihydro-1H-inden-1-yl, 2,3-dihydro-1H-inden-4-yl, benzo[d]isothiazol-3-yl, benzo[d]isoxazol-3-yl, 5-fluorobenzo[d]isoxazol-3-yl, 5-chlorobenzo[d]isoxazol-3-yl, 5-methylbenzo[d]isoxazol-3-yl, 1-methyl-2-oxo-1,2,3,4-tetrahydroquinolin-7-yl, 1-ethyl-1,2,3,4-tetrahydroquinolin-7-yl, 1-methyl-1,2,3,4-tetrahydroquinolin-5-yl, 1-(2,2,2-trifluoroethyl)-1,2,3,4-tetrahydroquinolin-7-yl, 5,6,7,8-tetrahydroquinolin-1-yl, 5,6,7,8-tetrahydroquinolin-3-yl, 1,2,3,4-tetrahydroquinolin-7-yl, 1-(oxetan-3-yl)-1,2,3,4-tetrahydroquinolin-7-yl, 1-(cyclopropylmethyl)-1,2,3,4-tetrahydroquinolin-7-yl, 1-methyl-1,2,3,4-tetrahydroquinolin-7-yl, 1-cyclopropyl-1,2,3,4-tetrahydroquinolin-7-yl, 1-acetyl-1,2,3,4-tetrahydroquinolin-7-yl, 1-oxo-3,4-dihydroquinolin-7-yl, 5,6,7,8-tetrahydro-1H-isoquinolin-7-yl, 2-ethyl-1,2,3,4-tetrahydro-1H-isoquinolin-8-yl, 2-ethyl-1,2,3,4-tetrahydro-1H-isoquinolin-5-yl, isoquinolin-2-yl, 1H-indazol-4-yl, 1H-indazol-3-yl, 1-methyl-1H-indazol-3-yl, 6-methoxy-1H-indol-4-yl, 2-methyl-1H-indol-4-yl, 1H-indol-4-yl, 1H-indol-3-yl, 3-methyl-1H-indol-4-yl, 7-chloro-5-fluoro-1H-indol-4-yl, 6-fluoro-1H-indol-4-yl, 1-(2-hydroxyethyl)-1H-indol-4-yl, 1-acetyl-1H-indol-4-yl, 1-cyclopropyl-1H-indol-4-yl, 6-fluoro-1-methyl-1H-indol-4-yl, 1-cyclopropyl-6-fluoro-1H-indol-4-yl, 1-methyl-1H-indol-4-yl, 3-fluoro-1-methyl-1H-indol-6-yl, 4-fluoro-1-methyl-1H-indol-6-yl, 7-chloro-5-fluoro-1-methyl-1H-indol-4-yl, 4-chloro-1-methyl-1H-indol-6-yl, 1-methylindolin-4-yl, 2,3-dihydrobenzofuran-4-yl, 1H-pyrrolo[3,2-c]pyridin-4-yl, 1H-pyrrolo[3,2-c]pyridin-3-yl, 4-fluoro-2-methyl-1H-indol-5-yl, 1-methyl-1H-pyrrolo[2,3-b]pyridin-4-yl, 1-methyl-1H-indazol-4-yl, 4-fluoro-1,2-dimethyl-1H-indol-5-yl, 1-cyclopropyl-4-fluoro-1H-indol-6-yl, 6-fluoro-1-(methyl-d3)-1H-indol-4-yl, 7-fluoro-2-methyl-1H-indol-4-yl, or 2-methyl-1H-indol-6-yl.
Provided herein is a compound of formula (II)
In some embodiments, R2 and R3 are each independent methoxy, fluoro, methoxy-d3, cyclopropoxy, fluoromethoxy, difluoromethoxy, or trifluoromethoxy. In some further embodiments, R2 is hydrogen and R3 is methoxy or methoxy-d3. In some further embodiments, R2 is methoxy or methoxy-d3 and R3 is methoxy or methoxy-d3. In some further embodiments, R2 is methoxy and R3 is methoxy-d3; or R2 is methoxy-d3 and R3 is methoxy; or R2 and R3 are each methoxy; or R2 and R3 are each methoxy-d3.
In some embodiments, X4 is N. In some embodiments, X4 is CH.
In some embodiments, R5 is —C(O)NR5aR5b, wherein R5a and R5b, together with the nitrogen atom to which they are attached, form a 4-, 5-, 6-, or 7-membered heterocyclyl ring comprising 0 or 1 additional heteroatom independently selected from nitrogen, oxygen or optionally oxidized sulfur as ring member, said ring is unsubstituted or substituted with one or two substituents R, wherein R is halogen, deuterium, C1-4alkyl, deuterated C1-4alkyl or —ORa, wherein Ra is hydrogen or C1-4alkyl deuterated C1-4alkyl; preferably fluoro, bromo, chloro, methyl, methyl-d3, ethyl, propyl, isopropyl, butyl, isobutyl, or tertbutyl. In some further embodiments, R5 is —C(O)NR5aR5b, wherein R5a and R5b, together with the nitrogen atom to which they are attached, form a 4-, 5-, 6-, or 7-membered heterocyclyl ring, which is azetidine-1-yl, pyrrolidine-1-yl, piperidine-1-yl, piperazine-1-yl, morpholine-4-yl, each of said ring is unsubstituted or substituted with one or two fluoro, bromo, chloro, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, or tertbutyl; preferably fluoro, or methyl. In some further embodiments, R5 is —C(O)NR5aR5b, wherein R5a and R5b, together with the nitrogen atom to which they are attached, form a 5- or 6-membered heterocyclyl ring, which is pyrrolidine-1-yl, or piperidine-1-yl, each of said ring is unsubstituted or substituted with fluoro. In some even further embodiments, R5 is 3,3-difluoropyrrolidine-1-carbonyl, piperidine-1-carbonyl, 4-fluoropiperidine-1-carbonyl, or 4,4-difluoropiperidine-1-carbonyl, preferably 4-fluoropiperidine-1-carbonyl.
In some embodiments, Cyc is indol-1-yl, indol-2-yl, indol-3-yl, indol-4-yl, indol-5-yl, indol-6-yl or indol-7-yl; or isoindol-1-yl, isoindol-2-yl, isoindol-3-yl, isoindol-4-yl, isoindol-5-yl, isoindol-6-yl or isoindol-7-yl, each of which is unsubstituted or substituted with one or two or three substituents selected from halogen, deuterium, C1-6alkyl, cyano, oxo, —ORd, —C(O)Rd, —C(O)NRdRe, —NRdRe, C3-8cycloalkyl, phenyl, heteroaryl, and heterocyclyl, each of said C1-6alkyl, C3-8cycloalkyl, phenyl, heteroaryl, and heterocyclyl is unsubstituted or substituted with halogen, deuterium, hydroxy, C1-6alkyl, C1-6alkoxy, or C3-8cycloalkyl, and wherein Rd and Re are each hydrogen, deuterated C1-6alkyl, C1-6alkyl, C3-8cycloalkyl, phenyl, heteroaryl, or heterocyclyl. In some further embodiments, Cyc is indol-4-yl, wherein position 6 of the indolyl ring is substituted with halogen (preferably fluoro) and position 1 of the indolyl ring is unsubstituted or substituted with C1-6alkyl (preferably methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tertbutyl), deuterated C1-6alkyl, C3-8cycloalkyl (preferably cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl), or hydoxyC1-6alkyl (preferably hydoxymethyl, hydoxyethyl, hydoxypropyl, or hydoxybutyl), and the other position is unsubstituted or substituted with halogen, C1-6alkyl or deuterated C1-6alkyl. In some embodiments, Cyc is indol-4-yl, wherein position 6 of the indolyl ring is unsubstituted and position 1 of the indolyl ring is substituted with C3-8cycloalkyl (preferably cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl), hydoxyC1-6alkyl (preferably hydoxymethyl, hydoxyethyl, hydoxypropyl, or hydoxybutyl) or —C(O)Rd, wherein Rd is hydrogen, C1-4alkyl, or C3-8cycloalkyl. In some embodiments, Cyc is indol-5-yl, wherein position 4 of the indolyl ring is substituted with halogen (preferably fluoro) and position 2 of the indolyl ring is unsubstituted or substituted with C1-6alkyl (preferably methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tertbutyl), deuterated C1-6alkyl, C3-8cycloalkyl (preferably cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl), or hydoxyC1-6alkyl (preferably hydoxymethyl, hydoxyethyl, hydoxypropyl, or hydoxybutyl), and the other position is unsubstituted or substituted with halogen, C1-6alkyl or deuterated C1-6alkyl. In some embodiments, Cyc is indol-5-yl, wherein position 4 of the indolyl ring is unsubstituted and position 1 of the indolyl ring is substituted with C3-8cycloalkyl (preferably cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl), hydoxyC1-6alkyl (preferably hydoxymethyl, hydoxyethyl, hydoxypropyl, or hydoxybutyl) or —C(O)Rd, wherein Rd is hydrogen, C1-4alkyl, or C3-8cycloalkyl. In some embodiments, Cyc is indol-6-yl, wherein position 4 of the indolyl ring is substituted with halogen (preferably fluoro) and position 1 of the indolyl ring is unsubstituted or substituted with C1-6alkyl (preferably methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tertbutyl), deuterated C1-6alkyl, C3-8cycloalkyl (preferably cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl), or hydoxyC1-6alkyl (preferably hydoxymethyl, hydoxyethyl, hydoxypropyl, or hydoxybutyl), and the other position is unsubstituted or substituted with halogen, C1-6alkyl or deuterated C1-6alkyl. In some embodiments, Cyc is indol-6-yl, wherein position 4 of the indolyl ring is unsubstituted and position 1 of the indolyl ring is substituted with C3-8cycloalkyl (preferably cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl), hydoxyC1-6alkyl (preferably hydoxymethyl, hydoxyethyl, hydoxypropyl, or hydoxybutyl) or —C(O)Rd, wherein Rd is hydrogen, C1-4alkyl, or C3-8cycloalkyl. In some embodiments, Cyc is indol-4-yl, indol-5-yl or indol-6-yl, wherein position 1 of the indolyl ring is substituted with C1-6alkyl (preferably methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tertbutyl), deuterated C1-6alkyl, C3-8cycloalkyl (preferably cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl), or hydoxyC1-6alkyl (preferably hydoxymethyl, hydoxyethyl, hydoxypropyl, or hydoxybutyl).
In some embodiments, Cyc is unsubstituted or substituted with one or two or three substituents selected from halogen, deuterium, cyano, C1-6alkyl, oxo, —ORd, —C(O)Ra, C3-8cycloalkyl, or heterocyclyl, said C1-6alkyl is unsubstituted or substituted with halogen, deuterium, hydroxy, or C3-6cycloalkyl, wherein Rd is hydrogen, C1-4alkyl, deuterated C1-4alkyl or C3-6cycloalkyl. In some embodiments, Cyc is unsubstituted or substituted with one or two or three substituents selected from fluoro, chloro, methyl, ethyl, isopropyl, n-propyl, methyl-d3, oxo, trifluoromethyl, oxetan-3-yl, cyclopropylmethyl, cyclopropyl, acetyl, methoxy, methoxy-d3 or 2-hydroxyethyl.
In some embodiments, Cyc is 6-methoxy-1H-indol-4-yl, 2-methyl-1H-indol-4-yl, 1H-indol-4-yl, 1H-indol-3-yl, 3-methyl-1H-indol-4-yl, 7-chloro-5-fluoro-1H-indol-4-yl, 6-fluoro-1H-indol-4-yl, 1-(2-hydroxyethyl)-1H-indol-4-yl, 1-acetyl-1H-indol-4-yl, 1-cyclopropyl-1H-indol-4-yl, 6-fluoro-I-methyl-1H-indol-4-yl, 1-cyclopropyl-6-fluoro-1H-indol-4-yl, 1-methyl-1H-indol-4-yl, 3-fluoro-1-methyl-1H-indol-6-yl, 4-fluoro-1-methyl-1H-indol-6-yl, 7-chloro-5-fluoro-I-methyl-1H-indol-4-yl, 4-chloro-1-methyl-1H-indol-6-yl, 1-methylindolin-4-yl, 4-fluoro-2-methyl-1H-indol-5-yl, 1-methyl-1H-pyrrolo[2,3-b]pyridin-4-yl, 1-methyl-1H-indazol-4-yl, 4-fluoro-1,2-dimethyl-1H-indol-5-yl, 1-cyclopropyl-4-fluoro-1H-indol-6-yl, 6-fluoro-1-(methyl-d3)-1H-indol-4-yl, 7-fluoro-2-methyl-1H-indol-4-yl, 2-methyl-1H-indol-6-yl, 6-methoxy-d3-11H-indol-4-yl, 2-methyl-d3-1H-indol-4-yl, 3-methyl-d3-1H-indol-4-yl, 1-methyl-d3-carbonyl-1H-indol-4-yl, 6-fluoro-1-methyl-d3-1H-indol-4-yl, 1-methyl-d3-1H-indol-4-yl, 3-fluoro-I-methyl-d3-1H-indol-6-yl, 4-fluoro-1-methyl-d3-1H-indol-6-yl, 7-chloro-5-fluoro-1-methyl-d3-1H-indol-4-yl, 4-chloro-1-methyl-d3-1H-indol-6-yl, 1-methyl-d3-indolin-4-yl, 4-fluoro-2-methyl-d3-1H-indol-5-yl, 1-methyl-d3-1H-pyrrolo[2,3-b]pyridin-4-yl, 1-methyl-d3-1H-indazol-4-yl, 4-fluoro-1,2-di(methyl-d3)-1H-indol-5-yl, 6-fluoro-1-(methyl-d3)-1H-indol-4-yl, 7-fluoro-2-methyl-d3-1H-indol-4-yl, or 2-methyl-d3-1H-indol-6-yl.
In alternative embodiments, the compound is selected from
Provided is a pharmaceutical composition comprising a compound disclosed herein or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, optionally together with a pharmaceutically acceptable excipient.
Provided is a method of treating a disease mediated by LPAR5, comprising administering a subject in need thereof a compound disclosed herein or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof; or a compound disclosed herein or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof for use in treating a disease mediated by LPAR5; or use of a compound disclosed herein or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for treating a disease mediated by LPAR5. In some embodiments, the disease mediated by LPAR5 is pain sensitizers-induced allodynia, neuropathic pain, and inflammatory pain, cancer, neurological disorders, atherosclerosis and cardiovascular disease, inflammatory and autoimmune diseases, fibrosis, bone development and disease, reproductive system and infertility, or obesity.
The following terms have the indicated meanings throughout the specification:
Unless specifically defined elsewhere in this document, all other technical and scientific terms used herein have the meaning commonly understood by one of ordinary skill in the art to which this invention belongs.
The following terms have the indicated meanings throughout the specification:
As used herein, including the appended claims, the singular forms of words such as “a”, “an”, and “the”, include their corresponding plural references unless the context clearly indicates otherwise.
The term “or” is used to mean, and is used interchangeably with, the term “and/or” unless the context clearly dictates otherwise.
The term “deuterated” is used herein to modify a chemical structure or an organic group or radical, wherein one or more carbon-bound hydrogen(s) are replaced by one or more deuterium(s), e.g., “deuterated-alkyl”, “deuterated-cycloalkyl” and the like. For example, the term “deuterated-alkyl” defined above refers to an alkyl group as defined herein, wherein at least one hydrogen atom bound to carbon is replaced by a deuterium. In a deuterated alkyl group, at least one carbon atom is bound to a deuterium; and it is possible for a carbon atom to be bound to more than one deuterium; it is also possible that more than one carbon atom in the alkyl group is bound to a deuterium.
The term “alkyl” includes a hydrocarbon group selected from linear and branched, saturated hydrocarbon groups comprising from 1 to 18, such as from 1 to 12, further such as from 1 to 10, more further such as from 1 to 8, or from 1 to 6, or from 1 to 4, carbon atoms. Examples of alkyl groups comprising from 1 to 6 carbon atoms (i.e., C1-6 alkyl) include, but not limited to, methyl, ethyl, 1-propyl or n-propyl (“n-Pr”), 2-propyl or isopropyl (“i-Pr”), 1-butyl or n-butyl (“n-Bu”), 2-methyl-1-propyl or isobutyl (“i-Bu”), 1-methylpropyl or s-butyl (“s-Bu”), 1,1-dimethylethyl or t-butyl (“t-Bu”), 1-pentyl, 2-pentyl, 3-pentyl, 2-methyl-2-butyl, 3-methyl-2-butyl, 3-methyl-1-butyl, 2-methyl-1-butyl, 1-hexyl, 2-hexyl, 3-hexyl, 2-methyl-2-pentyl, 3-methyl-2-pentyl, 4-methyl-2-pentyl, 3-methyl-3-pentyl, 2-methyl-3-pentyl, 2,3-dimethyl-2-butyl and 3,3-dimethyl-2-butyl groups. In some embodiments, an alkyl group, unless indicated otherwise, is optionally partially or fully deuterated, e.g., methyl-d3 or methoxy-d3.
The term “halogen” includes fluoro (F), chloro (Cl), bromo (Br) and iodo (I).
The term “haloalkyl” includes an alkyl group in which one or more hydrogen is/are replaced by one or more halogen atoms such as fluoro, chloro, bromo, and iodo. Examples of the haloalkyl include haloC1-8alkyl, haloC1-6alkyl or halo C1-4alkyl, but not limited to —CF3, —CH2C1, —CH2CF3, —CHCl2, —CF3, and the like.
The term “cycloalkyl” includes a hydrocarbon group selected from saturated cyclic hydrocarbon groups, comprising monocyclic and polycyclic (e.g., bicyclic and tricyclic) groups including fused, bridged or spiro cycloalkyl.
For example, the cycloalkyl group may comprise from 3 to 12, such as from 3 to 10, further such as 3 to 8, further such as 3 to 6, 3 to 5, or 3 to 4 carbon atoms. Even further for example, the cycloalkyl group may be selected from monocyclic group comprising from 3 to 12, such as from 3 to 10, further such as 3 to 8, 3 to 6 carbon atoms. Examples of the monocyclic cycloalkyl group include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl, cycloundecyl, and cyclododecyl groups. In particular, Examples of the saturated monocyclic cycloalkyl group, e.g., C3-8cycloalkyl, include, but not limited to cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl groups. In a preferred embedment, the cycloalkyl is a monocyclic ring comprising 3 to 6 carbon atoms (abbreviated as C3-6 cycloalkyl), including but not limited to, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl. Examples of the bicyclic cycloalkyl groups include those having from 7 to 12 ring atoms arranged as a fused bicyclic ring selected from [4,4], [4,5], [5,5], [5,6] and [6,6] ring systems, or as a bridged bicyclic ring selected from bicyclo[2.2.1]heptane, bicyclo[2.2.2]octane, and bicyclo[3.2.2]nonane. Further Examples of the bicyclic cycloalkyl groups include those arranged as a bicyclic ring selected from [5,6] and [6,6]ring systems.
The term “heteroaryl” includes a group selected from:
When the total number of S and O atoms in the heteroaryl group exceeds 1, those heteroatoms are not adjacent to one another. In some embodiments, the total number of S and O atoms in the heteroaryl group is not more than 2. In some embodiments, the total number of S and O atoms in the aromatic heterocycle is not more than 1. When the heteroaryl group contains more than one heteroatom ring member, the heteroatoms may be the same or different. The nitrogen atoms in the ring(s) of the heteroaryl group can be oxidized to form N-oxides.
Examples of the heteroaryl group or the monocyclic or bicyclic aromatic heterocyclic ring include, but are not limited to, (as numbered from the linkage position assigned priority 1) pyridyl (such as 2-pyridyl, 3-pyridyl, or 4-pyridyl), cinnolinyl, pyrazinyl, 2,4-pyrimidinyl, 3,5-pyrimidinyl, 2,4-imidazolyl, imidazopyridinyl, isoxazolyl, oxazolyl, thiazolyl, isothiazolyl, thiadiazolyl (such as 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, or 1,3,4-thiadiazolyl), tetrazolyl, thienyl (such as thien-2-yl, thien-3-yl), triazinyl, benzothienyl, furyl or furanyl, benzofuryl, benzoimidazolyl, indolyl (such as indol-1-yl, indol-2-yl, indol-3-yl, indol-4-yl, indol-5-yl, indol-6-yl or indol-7-yl), isoindolyl (such as isoindol-1-yl, isoindol-2-yl, isoindol-3-yl, isoindol-4-yl, isoindol-5-yl, isoindol-6-yl or isoindol-7-yl), indolinyl, oxadiazolyl (such as 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, or 1,3,4-oxadiazolyl), phthalazinyl, pyrazinyl, pyridazinyl, pyrrolyl, triazolyl (such as 1,2,3-triazolyl, 1,2,4-triazolyl, or 1,3,4-triazolyl), quinolinyl, isoquinolinyl, pyrazolyl, pyrrolopyridinyl (such as 1H-pyrrolo[2,3-b]pyridin-5-yl), pyrazolopyridinyl (such as 1H-pyrazolo[3,4-b]pyridin-5-yl), benzofuranyl, benzoxazolyl (such as benzo[d]oxazol-6-yl), pteridinyl, purinyl, 1-oxa-2,3-diazolyl, 1-oxa-2,4-diazolyl, 1-oxa-2,5-diazolyl, 1-oxa-3,4-diazolyl, 1-thia-2,3-diazolyl, 1-thia-2,4-diazolyl, 1-thia-2,5-diazolyl, 1-thia-3,4-diazolyl, furazanyl (such as furazan-2-yl, furazan-3-yl), benzofurazanyl, benzothiophenyl, benzothiazolyl, benzoxazolyl, quinazolinyl, quinoxalinyl, naphthyridinyl, furopyridinyl, benzothiazolyl (such as benzo[d]thiazol-6-yl), indazolyl (such as 1H-indazol-5-yl) and 5,6,7,8-tetrahydroisoquinoline.
“Heterocyclyl”, “heterocycle” or “heterocyclic” are interchangeable and include a non-aromatic heterocyclyl group comprising one or more, e.g., 1 to 3, heteroatoms selected from nitrogen, oxygen or optionally oxidized sulfur as ring members, with the remaining ring members being carbon, including monocyclic, fused, bridged, and spiro ring, i.e., containing monocyclic heterocyclyl, bridged heterocyclyl, spiro heterocyclyl, and fused heterocyclic groups.
Exemplary monocyclic 4 to 9-membered heterocyclyl groups include, but not limited to, (as numbered from the linkage position assigned priority 1) pyrrolidin-1-yl, pyrrolidin-2-yl, pyrrolidin-3-yl, imidazolidin-2-yl, imidazolidin-4-yl, pyrazolidin-2-yl, pyrazolidin-3-yl, piperidin-1-yl, piperidin-2-yl, piperidin-3-yl, piperidin-4-yl, 2,5-piperazinyl, pyranyl, morpholinyl, morpholino, morpholin-2-yl, morpholin-3-yl, oxiranyl, aziridin-1-yl, aziridin-2-yl, azocan-1-yl, azocan-2-yl, azocan-3-yl, azocan-4-yl, azocan-5-yl, thiiranyl, azetidin-1-yl, azetidin-2-yl, azetidin-3-yl, oxetanyl, thietanyl, 1,2-dithietanyl, 1,3-dithietanyl, dihydropyridinyl, tetrahydropyridinyl, thiomorpholinyl, thioxanyl, piperazinyl, homopiperazinyl, homopiperidinyl, azepan-1-yl, azepan-2-yl, azepan-3-yl, azepan-4-yl, oxepanyl, thiepanyl, 1,4-oxathianyl, 1,4-dioxepanyl, 1,4-oxathiepanyl, 1,4-oxaazepanyl, 1,4-dithiepanyl, 1,4-thiazepanyl and 1,4-diazepanyl, 1,4-dithianyl, 1,4-azathianyl, oxazepinyl, diazepinyl, thiazepinyl, dihydrothienyl, dihydropyranyl, dihydrofuranyl, tetrahydrofuranyl, tetrahydrothienyl, tetrahydropyranyl, tetrahydrothiopyranyl, 1-pyrrolinyl, 2-pyrrolinyl, 3-pyrrolinyl, indolinyl, 2H-pyranyl, 4H-pyranyl, 1,4-dioxanyl, 1,3-dioxolanyl, pyrazolinyl, pyrazolidinyl, dithianyl, dithiolanyl, pyrazolidinyl, imidazolinyl, pyrimidinonyl, or 1,1-dioxo-thiomorpholinyl.
The term “stereoisomer” refers to all isomers of individual compounds that differ only in the orientation of their atoms in space. The term stereoisomer includes mirror image isomers (enantiomers), mixtures of mirror image isomers (racemates, racemic mixtures), geometric (cis/trans or syn/anti or E/Z) isomers, and isomers of compounds with more than one chiral center that are not mirror images of one another (diastereoisomers).
Compounds disclosed herein may contain an asymmetric center and may thus exist as enantiomers. “Enantiomers” refer to two stereoisomers of a compound which are non-superimposable mirror images of one another. Where the compounds disclosed herein possess two or more asymmetric centers, they may additionally exist as diastereomers. Enantiomers and diastereomers fall within the broader class of stereoisomers. All such possible stereoisomers as substantially pure resolved enantiomers, racemic mixtures thereof, as well as mixtures of diastereomers are intended to be included. All stereoisomers of the compounds disclosed herein and/or pharmaceutically acceptable salts thereof are intended to be included. Unless specifically mentioned otherwise, reference to one isomer applies to any of the possible isomers. Whenever the isomeric composition is unspecified, all possible isomers are included.
When compounds disclosed herein contain olefinic double bonds, unless specified otherwise, such double bonds are meant to include both E and Z geometric isomers.
When compounds disclosed herein contain a di-substituted cyclic ring system, substituents found on such ring system may adopt cis and trans formations. Cis formation means that both substituents are found on the upper side of the 2 substituent placements on the carbon, while trans would mean that they were on opposing sides. For example, the di-substituted cyclic ring system may be cyclohexyl or cyclobutyl ring.
It may be advantageous to separate reaction products from one another and/or from starting materials. The desired products of each step or series of steps is separated and/or purified (hereinafter separated) to the desired degree of homogeneity by the techniques common in the art. Typically such separations involve multiphase extraction, crystallization from a solvent or solvent mixture, distillation, sublimation, or chromatography. Chromatography can involve any number of methods including, for example: reverse-phase and normal phase; size exclusion; ion exchange; high, medium and low pressure liquid chromatography methods and apparatus; small scale analytical; simulated moving bed (“SMB”) and preparative thin or thick layer chromatography, as well as techniques of small scale thin layer and flash chromatography. One skilled in the art could select and apply the techniques most likely to achieve the desired separation.
“Diastereomers” refer to stereoisomers of a compound with two or more chiral centers but which are not mirror images of one another. Diastereomeric mixtures can be separated into their individual diastereomers on the basis of their physical-chemical differences by methods well known to those skilled in the art, such as by chromatography and/or fractional crystallization. Enantiomers can be separated by converting the enantiomeric mixture into a diastereomeric mixture by reaction with an appropriate optically active compound (e.g., chiral auxiliary such as a chiral alcohol or Mosher's acid chloride), separating the diastereomers and converting (e.g., hydrolyzing) the individual diastereoisomers to the corresponding pure enantiomers. Enantiomers can also be separated by the use of a chiral HPLC column.
A single stereoisomer, e.g., a substantially pure enantiomer, may be obtained by resolution of the racemic mixture using a method such as the formation of diastereomers using optically active resolving agents (Eliel, E. and Wilen, S. Stereochemistry of Organic Compounds. New York: John Wiley & Sons, Inc., 1994; Lochmuller, C. H., et al. “Chromatographic resolution of enantiomers: Selective review.” J. Chromatogr., 113(3) (1975): pp. 283-302). Racemic mixtures of chiral compounds of the invention can be separated and isolated by any suitable method, including (1) formation of ionic, diastereomeric salts with chiral compounds and separation by fractional crystallization or other methods, (2) formation of diastereomeric compounds with chiral derivatizing reagents, separation of the diastereomers, and conversion to the pure stereoisomers, and (3) separation of the substantially pure or enriched stereoisomers directly under chiral conditions. See: Wainer, Irving W., Ed. Drug Stereochemistry: Analytical Methods and Pharmacology. New York: Marcel Dekker, Inc., 1993.
“Pharmaceutically acceptable salts” refer to those salts which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response, and the like, and are commensurate with a reasonable benefit/risk ratio. A pharmaceutically acceptable salt may be prepared in situ during the final isolation and purification of the compounds disclosed herein, or separately by reacting the free base function with a suitable organic acid or by reacting the acidic group with a suitable base.
In addition, if a compound disclosed herein is obtained as an acid addition salt, the free base can be obtained by basifying a solution of the acid salt. Conversely, if the product is a free base, an addition salt, such as a pharmaceutically acceptable addition salt, may be produced by dissolving the free base in a suitable organic solvent and treating the solution with an acid, in accordance with conventional procedures for preparing acid addition salts from base compounds. Those skilled in the art will recognize various synthetic methodologies that may be used without undue experimentation to prepare non-toxic pharmaceutically acceptable addition salts.
As defined herein, “a pharmaceutically acceptable salt thereof” includes salts of at least one compound of Formula (I), and salts of the stereoisomers of the compound of Formula (I), such as salts of enantiomers, and/or salts of diastereomers.
The terms “administration”, “administering”, “treating” and “treatment” herein, when applied to an animal, human, experimental subject, cell, tissue, organ, or biological fluid, mean contact of an exogenous pharmaceutical, therapeutic, diagnostic agent, or composition to the animal, human, subject, cell, tissue, organ, or biological fluid. Treatment of a cell encompasses contact of a reagent to the cell, as well as the contact of a reagent to a fluid, where the fluid is in contact with the cell. The term “administration” and “treatment” also means in vitro and ex vivo treatments, e.g., of a cell, by a reagent, diagnostic, binding compound, or by another cell. The term “subject” herein includes any organism, preferably an animal, more preferably a mammal (e.g., rat, mouse, dog, cat, and rabbit) and most preferably a human.
The term “effective amount” or “therapeutically effective amount” refers to an amount of the active ingredient, such as a compound that, when administered to a subject for treating a disease, or at least one of the clinical symptoms of a disease or disorder, is sufficient to effect such treatment for the disease, disorder, or symptom. The term “therapeutically effective amount” can vary with the compound, the disease, disorder, and/or symptoms of the disease or disorder, the severity of the disease, disorder, and/or symptoms of the disease or disorder, the age of the subject to be treated, and/or the weight of the subject to be treated. An appropriate amount in any given instance can be apparent to those skilled in the art or can be determined by routine experiments. In some embodiments, “therapeutically effective amount” is an amount of at least one compound and/or at least one stereoisomer thereof, and/or at least one pharmaceutically acceptable salt thereof disclosed herein effective to “treat” as defined herein, a disease or disorder in a subject. In the case of combination therapy, the term “therapeutically effective amount” refers to the total amount of the combination objects for the effective treatment of a disease, a disorder or a condition.
The term “disease” refers to any disease, discomfort, illness, symptoms or indications, and can be interchangeable with the term “disorder” or “condition”.
Throughout this specification and the claims which follow, unless the context requires otherwise, the term “comprise”, and variations such as “comprises” and “comprising” are intended to specify the presence of the features thereafter, but do not exclude the presence or addition of one or more other features. When used herein the term “comprising” can be substituted with the term “containing”, “including” or sometimes “having”.
Throughout this specification and the claims which follow, the term “Cn-m” indicates a range which includes the endpoints, wherein n and m are integers and indicate the number of carbons. Examples include C1-8, C1-6, and the like.
The term “at least one substituent” disclosed herein includes, for example, from 1 to 4, such as from 1 to 3, further as 1 or 2, substituents, provided that the theory of valence is met. For example, “at least one substituent R” disclosed herein includes from 1 to 4, such as from 1 to 3, further as 1 or 2, substituents selected from the list of R as disclosed herein.
To a mixture of 2-bromo-4,5-dimethoxybenzoic acid (25 g, 95.7 mmol), CuBr (1.37 g, 9.58 mmol) and diethyl malonate (210 g, 1311 mmol) at 0° C. was added NaH (9.19 g, 229.8 mmol, 60% in oil) portion wise. After addition, the reaction mixture was heated to 90° C. for 2 hours. After cooling to RT, water (1.0 L) was added and the whole was extracted with PE (600 mL×2). The aqueous layer was acidified to pH 6 with aq. HCl (6.0 M) and extracted with EtOAc (600 mL×2). The combined organic layers were dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to afford the title compound (29.8 g, 80.97 mmol, 84.6% yield) as a yellow solid. LC-MS: calc. for [C16H21O8]+: 341.12, found: 341.0 [M+H]+
A mixture of 2-(1,3-diethoxy-1,3-dioxopropan-2-yl)-4,5-dimethoxybenzoic acid (28.8 g, 84.624 mmol,) in EtOH (400 mL) and con. HCl (100 mL) was heated to 100° C. for 16 hours. The resulting mixture was concentrated under reduced pressure. The residue was mixed with EtOH (200 mL). and the resulting mixture was concentrated under reduced pressure twice. The residue was mixed with EtOH (400 mL) and the whole was cooled to 0° C., then SOCl2 (40 mL) was added dropwise. The resulting mixture was heated to reflux for 16 hours and then concentrated under reduced pressure. The residue was diluted with water (400 mL) and the whole was extracted with EtOAc (400 mL×3). The organic layers were combined, washed with brine (300 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was slurried by PE/EtOAc (10/1, 330 mL) to afford the title compound (20 g, 64.66 mmol, 76.4% yield) as a yellow solid. LC-MS: calc. for [C15H21O6]+:297.13, found: 297.3[M+H]+
A mixture of ethyl 2-(2-ethoxy-2-oxoethyl)-4,5-dimethoxybenzoate (140 mg, 0.472 mmol) and [(tert-butoxy)(dimethylamino)methyl]dimethylamine (0.20 mL, 0.945 mmol) was stirred at 100° C. for 3 h. After cooling to rt, to the resulting mixture were added acetic acid (8.0 mL) and benzo[d]isothiazol-3-amine (213 mg, 1.42 mmol). The reaction was stirred at RT for 16 hr. The resulting mixture was concentrated under reduced pressure to afford the crude product (200 mg, 0.31 mmol, 65.0% yield) as a yellow oil. LC-MS: calc. for [C21H19N2O5S]+:411.09, found: 411.0 [M+H]+
To a solution of ethyl 2-(benzo[d]isothiazol-3-yl)-6,7-dimethoxy-1-oxo-1,2-dihydroisoquinoline-4-carboxylate (100 mg, 0.244 mmol) in MeOH (6 mL) was added LiOH H2O (102 mg, 2.44 mmol), and the reaction mixture was stirred at 70° C. for 2 hr. The mixture was cooled to RT and adjusted to pH 7˜8 with aq. HCl (1.0 M). The resulting mixture was concentrated under reduced pressure to remove most of MeOH. The residue was adjusted to pH 3-4 with aq. HCl (1.0 M). The whole was filtered in a vacuum. The filter cake was collected, dried in a vacuum to afford the crude title compound (30 mg, 0.020 mmol, 10.0% yield) as a white solid. LC-MS: calc. for [C19H15N2O5S]+:383.06, found: 381.0 [M+H]+
To a solution of 2-(benzo[d]isothiazol-3-yl)-6,7-dimethoxy-1-oxo-1,2-dihydroisoquinoline-4-carboxylic acid (30 mg, 0.073 mmol) in DMF (2.0 mL) were added piperidine (0.030 mL, 0.328 mmol), DIEA (0.05 mL, 0.328 mmol) and HATU (37.4 mg, 0.098 mmol), and the reaction was stirred at room temperature for 1 hr. The reaction mixture was purified by Pre-HPLC (Base method: NH3·H2O in CH3CN) to afford the title compound (14.04 mg, 0.03 mmol, 39.8% yield). LC-MS: calc. for [C24H24N3O4S]+:450.14, found: 450.1 [M+H]+. 1H NMR (400 MHz, DMSO) δ 8.48 (s, 1H), 8.32 (d, J=8.3 Hz, 1H), 7.85 (d, J=8.2 Hz, 1H), 7.68-7.73 (m, 1H), 7.60 (s, 1H), 7.51-7.55 (m, 1H), 6.95 (s, 1H), 3.92 (s, 3H), 3.40-3.55 (m, 4H), 1.40-1.60 (m, 6H).
The compounds below were synthesized following the procedures described for Compound 1
1H NMR (400 MHz, DMSO-d6) δ
A mixture of ethyl 2-(2-ethoxy-2-oxoethyl)-4,5-dimethoxybenzoate (644 mg, 2.17 mmol) and [(tert-butoxy)(dimethylamino)methyl]dimethylamine (0.90 mL, 4.35 mmol) was stirred at 100° C. for 2 hr. The mixture was cooled to rt, then HOAc (15 mL) and 5,6,7,8-tetrahydronaphthalen-2-amine (960 mg, 6.52 mmol) were added. Then the reaction mixture was stirred at 50° C. for 18 hr. The mixture was diluted with water (60 mL) and the resulting mixture was extracted with EA (50 mL*2). The organic layers were combined, washed with brine (50 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE/EA=20/1 to 3/1) to afford the title compound (605 mg, 1.33 mmol, 61.4% yield) as a yellow solid. LC-MS (ESI): m/z 454.2 [M+H]+
A mixture of ethyl 2-(3-ethoxy-3-oxo-1-((5,6,7,8-tetrahydronaphthalen-2-yl)amino)prop-1-en-2-yl)-4,5-dimethoxybenzoate (605 mg, 1.33 mmol), LiOH·H2O (280 mg, 6.67 mmol), MeOH (12 mL) and H2O (3 mL) was stirred at 70° C. for 3 hr. The reaction mixture was cooled to rt, then adjusted to pH=10-11 with diluted HCl (1.0 M). The resulting suspension was filtered in a vacuum to afford the crude title compound (659 mg, 1.74 mmol, 77.8% yield) as a white solid, which was used in the next step without further purification. LC-MS (ESI): m/z 380.1 [M+H]+
A mixture of 6,7-dimethoxy-1-oxo-2-(5,6,7,8-tetrahydronaphthalen-2-yl)-1,2-dihydroisoquinoline-4-carboxylic acid (100 mg, 0.26 mmol), DIEA (0.13 mL, 0.79 mmol), HATU (120 mg, 0.32 mmol) and DMF (6 mL) was stirred at room temperature for 30 min before piperidine (0.030 mL, 0.32 mmol) was added. The reaction mixture was stirred at room temperature for 1.5 hr. The mixture was diluted with water (60 mL) and the resulting mixture was extracted with EA (50 mL*2). The organic layers were combined, washed with brine (50 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE/EA=10/1 to 1/2) to afford the title compound (86 mg, 0.19 mmol, 73.1% yield). LC-MS (ESI): m/z 447.2 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ 7.67 (s, 1H), 7.35 (s, 1H), 7.21-7.15 (m, 2H), 7.14 (s, 1H), 6.91 (s, 1H), 3.88 (d, J=2.5 Hz, 6H), 3.74-3.31 (m, 4H), 2.77 (d, J=4.3 Hz, 4H), 1.79-1.75 (m, 4H), 1.63-1.44 (m, 6H).
The compounds below were synthesized following the procedures described for Compound 16
1H NMR (400 MHz, DMSO-d6) δ
To a mixture of (2,4-dimethoxyphenyl)methanamine and [(tert-butoxy)(dimethylamino) methyl]dimethylamine (8.27 mL, 39.87 mmol) was stirred at 100° C. for 3 h. The mixture was cooled to RT. AcOH (30 mL) and ethyl 2-(2-ethoxy-2-oxoethyl)-4,5-dimethoxybenzoate (9.01 mL, 59.8 mmol) were added. Then the reaction mixture was stirred at 50° C. overnight. The mixture was diluted with EA (250 mL), then washed with brine (200 mL*2). The organic layer was separated and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE/EA=10/1-1/1) to afford the title compound (8.60 g, 18.2 mmol, 91.1% yield).
To a solution of ethyl 2-(1-((2,4-dimethoxybenzyl)amino)-3-ethoxy-3-oxoprop-1-en-2-yl)-4,5-dimethoxybenzoate (8.60 g, 18.1 mmol) in MeOH (300 mL) were added H2O (100 mL) and LiOH·H2O (6.10 g, 145 mmol), and the reaction was stirred at 80° C. for 4 hr. The reaction mixture was cooled to rt, then adjusted to pH=2-3 with con HCl. The resulting mixture was filtered in a vacuum and the filter cake was collected, dried in vacuum to afford the title compound (6.20 g, 85.5% yield) as a white solid. LC-MS (ESI): m/z 400.0 [M+H]+
To a solution of 2-(2,4-dimethoxybenzyl)-6,7-dimethoxy-I-oxo-1,2-dihydroisoquinoline-4-carboxylic acid (6.20 g, 15.5 mmol) in DMF (60 mL) were added HATU (7.08 g, 18.6 mmol), TEA (6.50 mL, 46.6 mmol) and piperidine (3.10 mL, 31.0 mmol). The reaction was stirred at room temperature for 1 hr. The reaction mixture was diluted with brine (300 mL). The mixture was extracted with EA (150 mL*3). The organic layers were combined, dried over anhydrous Na2SO4, filtered and the filtrate was concentrated in a vacuum. The residue was purified by silica gel column chromatography (PE/EA=10/1 to 1/4) to afford the crude title compound (7.70 g) as a white solid, 100 mg of which was purified with Prep-HPLC (Waters 2767/2545/2489/Qda, Column name: Inertsil ODS-3 10 μm 20*250 nm, Mobile Phase A: 0.1% HCOOH in water, Mobile Phase B: CH3CN, Flow: 20 mL/min: Column temp: RT) to afford the title compound (32.7 mg, 32.7% yield) as a white solid. LC-MS (ESI): m/z 467.1 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ 7.64 (s, 1H), 7.38 (s, 1H), 7.03 (d, J=8.4 Hz, 1H), 6.87 (s, 1H), 6.60-6.57 (m, 1H), 6.49-6.46 (m, 1H), 5.03 (s, 2H), 3.87 (s, 3H), 3.85 (s, 3H), 3.80 (s, 3H), 3.74 (s, 3H), 3.55-3.35 (m, 4H), 1.58-1.40 (m, 6H).
A solution of 2-(2,4-dimethoxybenzyl)-6,7-dimethoxy-4-(piperidine-1-carbonyl)isoquinolin-1(2H)-one (7.60 g, 16.3 mmol) in TFA (75 mL) was heated to 85° C. and stirred at this temperature for 1 hr. The reaction was concentrated in a vacuum. The residue was purified by silica gel column chromatography (DCM/MeOH=100/1 to 10/1) to afford the title compound (3.2 g, 62.1% yield) as a light yellow solid. LC-MS (ESI): m/z 317.0 [M+H]+
A mixture of 6,7-dimethoxy-4-(piperidine-1-carbonyl)isoquinolin-1(2H)-one (100 mg, 0.32 mmol), 2-bromo-3-methylpyridine (115 mg, 0.63 mmol), CuI (3.0 mg, 0.016 mmol), K2CO3 (131 mg, 0.95 mmol), N1,N2-dimethylethane-1,2-diamine (2.8 mg, 0.03 mmol) and DMSO (3 mL) was bubbled with argon for 3 seconds, then microwaved at 150° C. for 2 hr. The reaction mixture was purified with prep-HPLC (Waters 2767/2545/2489/Qda, Column name: Inertsil ODS-3 10 m 20*250 nm, Mobile Phase A: 0.1% HCOOH in water, Mobile Phase B: CH3CN, Flow: 20 mL/min: Column temp: RT) to afford the title compound (12.4 mg, 9.3% yield). LC-MS (ESI): m/z 418.3 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ 8.06 (dd, J=7.6, 1.2 Hz, 1H), 9.91 (td, J=8.0, 1.6 Hz, 1H), 7.75-7.63 (m, 3H), 7.53 (s, 1H), 6.94 (s, 1H), 3.91 (s, 3H), 3.90 (s, 3H), 3.70-3.40 (m, 4H), 1.70-1.40 (m, 4H).
The compounds below were synthesized following the procedures described for Compound 36 and 37.
1H NMR (400 MHz, DMSO-d6) δ
A mixture of 6,7-dimethoxy-4-(piperidine-1-carbonyl)isoquinolin-1(2H)-one (130 mg, 0.410 mmol), 2-bromo-3-methylpyridine (141 mg, 0.82 mmol), CuI (4.0 mg, 0.02 mmol), K3PO4 (262 mg, 1.23 mmol), N1,N2-dimethylethane-1,2-diamine (3.6 mg, 0.04 mmol) and toluene (5 mL) was bubbled with argon for 12 seconds, then microwaved at 120° C. for 1 hr. The reaction mixture was purified with Prep-HPLC (Waters 2767/2545/2489/Qda, Column name: Inertsil ODS-3 10 m 20*250 nm, Mobile Phase A: 0.1% HCOOH in water, Mobile Phase B: CH3CN, Flow: 20 mL/min: Column temp: RT) to afford the title compound (10.58 mg, 6.3% yield). LC-MS:(ESI): m/z 408.1 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ 8.47-8.43 (m, 1H), 7.91-7.89 (d, J=6.4 Hz, 1H), 7.69 (s, 1H), 7.50-7.47 (m, 1H) 7.41 (s, 1H), 6.94 (s, 1H), 3.90 (s, 3H), 3.89 (s, 3H), 3.75-3.41 (m, 4H), 2.14 (s, 3H), 1.75-1.30 (m, 6H).
The compounds below were synthesized following the procedures described for Compound 50.
1H NMR (400 MHz, DMSO-d6) δ
To a solution of 7-nitro-1,2,3,4-tetrahydroquinoline (5.00 g, 28.1 mmol) in DCM (8 mL) were added DIEA (14.0 mL, 84.2 mmol) and acetyl chloride (3.00 mL, 42.1 mmol) consecutively at 0° C. under N2 atmosphere. The resulting reaction mixture was stirred at rt for 2 h. The mixture was diluted with water (50 mL), then extracted with DCM (50 mL*2). The organic layers were combined, washed with brine (20 mL), dried over anhydrous Na2SO4, filtered and concentrated. The residue was purified by silica gel column (EA/PE=1/50 to 3/1) to afford the titled compound (4.5 g, 20.43 mmol, 72.8% yield) as a yellow solid. LC-MS (ESI): m/z 221.2 [M+H]+
A mixture of 1-(7-nitro-3,4-dihydroquinolin-1(2H)-yl)ethan-1-one (4.00 g, 18.2 mmol), 10% Pd/C (1.93 g) and MeOH (50 mL) was hydrogenated with a balloon for 18 hours. The resulting mixture was filtered in vacuum. The filtrate was concentrated. The residue was purified by silica gel column (PE/EA=50/1 to 1/1) to afford the titled compound (3.73 g, 19.61 mmol, 107.9% yield) as a colorless oil. LC-MS (ESI): m/z 191.5 [M+H]+
A mixture of ethyl 2-(2-ethoxy-2-oxoethyl)-4,5-dimethoxybenzoate (1.42 g, 4.78 mmol) and 1-tert-butoxy-N,N,N′,N′-tetramethylmethanediamine (1.98 mL, 9.57 mmol) was stirred at 100° C. for 3 h. The mixture was cooled to rt, then HOAc (50 mL) and 1-(7-amino-3,4-dihydroquinolin-1(21H)-yl)ethan-1-one (2.73 g, 14.4 mmol) were added. Then the reaction mixture was stirred at 50° C. overnight. The resulting mixture was diluted with EA (20 mL), then washed with brine (30 mL*2). The organic layer was concentrated in vacuo. The residue was purified by silica gel column chromatography (PE/EA=10/1 to 1/1) to afford the title compound the titled compound (2.64 g, 5.32 mmol, 111.3% yield) as a yellow oil. LC-MS (ESI): m/z 497.1 [M+H]+
To a solution of ethyl 2-(1-((1-acetyl-1,2,3,4-tetrahydroquinolin-7-yl)amino)-3-ethoxy-3-[0119]oxoprop-1-en-2-yl)-4,5-dimethoxybenzoate (400 mg, 0.806 mmol) in MeOH (6 mL) and H2O (6 mL) was added LiOH·H2O (373 mg, 8.88 mmol). The mixture was heated to 80 C for 6 h. The reaction mixture was cooled to rt, then adjusted to pH=2-3 with con. HCl. The resulting mixture was concentrated in vacuo to remove MeOH. The residue was filtered in vacuum to afford the title compound the titled compound (200 mg, 0.53 mmol, 59.2% yield) as a brown solid. LC-MS (ESI): m/z 381.0 [M+H]+
To a solution of 6,7-dimethoxy-1-oxo-2-(1,2,3,4-tetrahydroquinolin-7-yl)-1,2-dihydroisoquinoline-4-carboxylic acid (300 mg, 0.79 mmol) in DMF (10 mL) were added DIEA (0.26 mL, 1.57 mmol), HATU (389.8 mg, 1.02 mmol) and piperidine (0.09 mL, 0.95 mmol). The reaction was stirred at room temperature for 2 h. The mixture was diluted with DCM (50 mL), washed with brine (50 mL). The organic layer was dried over anhydrous Na2SO4, then filtered in vacuum. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC (Waters 2767/2545/2489, Waters Xbridge C18 10 μm OBD 19*250 mm, Mobile Phase A: 0.1% FA in water, Mobile Phase B: CH3CN, Flow: 20 mL/min, Column temp: RT) to afford the titled compound (5.09 mg, 0.01 mmol, 1.4% yield) as a white solid. LC-MS (ESI): m/z 448.1 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ 7.66 (s, 1H), 7.27 (s, 1H), 6.94 (d, J=7.9 Hz, 1H), 6.90 (s, 1H), 6.45 (s, 1H), 6.42 (d, J=7.8 Hz, 1H), 5.93 (s, 1H), 3.88 (d, J=5.2 Hz, 6H), 3.52-3.40 (m, 4H), 3.21 (s, 2H), 2.71 (s, 2H), 1.82 (s, 2H), 1.60 (s, 3H), 1.50 (s, 3H).
A mixture of 6,7-dimethoxy-4-(piperidine-1-carbonyl)-2-(1,2,3,4-tetrahydroquinolin-7-yl)isoquinolin-1(2H)-one (100 mg, 0.22 mmol), oxetan-3-one (72.3 mg, 1.00 mmol), HOAc (0.01 mL, 0.07 mmol), NaCNBH3 (42.04 mg, 0.67 mmol) and DCE (10 mL) was degassed with argon for 3 times, then the reaction was stirred at 50° C. overnight. The mixture was diluted with DCM (50 mL), then washed with brine (50 mL). The organic layer was dried over anhydrous Na2SO4, filtered and concentrated. The residue was purified by Prep-HPLC (Waters 2767/2545/2489, Waters Xbridge C18 10 μm OBD 19*250 mm, Mobile Phase A: 0.10% FA in water, Mobile Phase B: CH3CN, Flow: 20 mL/min, Column temp: RT) to afford the titled compound (35.72 mg, 0.07 mmol, 31.7% yield). LC-MS (ESI): m/z 504.7 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ 7.67 (s, TH), 7.30 (s, 1H), 7.07 (d, J=7.9 Hz, 1H), 6.92 (s, 1H), 6.63 (dd, J=7.8, 1.9 Hz, 1H), 6.32 (d, J=1.9 Hz, 1H), 4.75-4.66 (m, 5H), 3.88 (d, J=5.4 Hz, 6H), 3.49 (s, 4H), 3.27-3.23 (m, 2H), 2.75 (t, J=6.4 Hz, 2H), 1.96-1.90 (m, 2H), 1.61-1.50 (m, 6H).
The compounds below were synthesized following the procedures described for Compound 60 and 61.
1H NMR (400 MHz, DMSO-d6) δ
A mixture of 6,7-dimethoxy-4-(piperidine-1-carbonyl)-2-(1,2,3,4-tetrahydroquinolin-7-yl)isoquinolin-1(2H)-one (50 mg, 0.11 mmol), acetyl chloride (0.01 mL, 0.17 mmol), K2CO3 (15.4 mg, 0.11 mmol) and DMF (3 mL) was degassed with argon for 3 times, then the reaction mixture was stirred at 100° C. overnight. The reaction mixture was diluted with EA (50 mL), then washed with brine (50 mL). The organic layer was dried over anhydrous Na2SO4, filtered and concentrated. The residue was purified by prep-HPLC (Waters 2767/2545/2489, Waters Xbridge C18 10 um OBD 19*250 mm, Mobile Phase A: 0.10% FA in water, Mobile Phase B: CH3CN, Flow: 20 mL/min, Column temp: RT) to afford the title compound (17.4 mg, 0.04 mmol, 31.8% yield). LC-MS (EST): m/z 490.1 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ 7.68 (s, 2H), 7.37 (s, 1H), 7.32 (d, J=8.2 Hz, 1H), 7.17 (dd, J=8.1, 2.0 Hz, 1H), 6.92 (s, 1H), 3.88 (d, J=3.7 Hz, 6H), 3.72 (t, J=6.2 Hz, 2H), 3.49 (s, 4H), 2.78 (t, J=6.4 Hz, 2H), 2.22 (s, 3H), 1.95-1.87 (m, 2H), 1.61-1.50 (m, 6H).
To a solution of 2-(1H-indol-4-yl)-6,7-dimethoxy-4-(piperidine-1-carbonyl)isoquinolin-1(2H)-one (180 mg, 0.42 mmol) in DMF (4 mL) were added (2-bromoethoxy)(tert-butyl)dimethylsilane (103 mg, 0.46 mmol), Cs2CO3 (272 mg, 0.83 mmol).The mixture reaction was stirred at 80° C. for 2 hr. The mixture reaction was diluted with H2O (50 mL) and extracted with EA (20 Ml*3). The organic layer was combined, washed with brine (50 mL), dried over anhydrous Na2SO4, filtered and concentrated to afford the crude title compound (180 mg, 0.24 mmol, 58.3% yield) as a yellow solid. LC-MS (ESI): m/z 590.3 [M+H]+.
A solution of 2-(1-(2-((tert-butyldimethylsilyl)oxy)ethyl)-1H-indol-4-yl)-6,7-dimethoxy-4-(piperidine-1-carbonyl)isoquinolin-1(2H)-one (180 mg, 0.31 mmol) and TBAF (5.0 mL, 5.0 mmol, 1 mol/L in THF) was stirred at RT for 2 hr. The mixture was purified by Prep-HPLC (system: Waters 2767/2545/2489/Qda, Waters sunfire C18 10 μm OBD 19*250 mm, Mobile Phase A: 0.1% FA in water, Mobile Phase B: CH3CN, Flow: 20 mL/min: Column temp: RT) to afford the title compound (70.0 mg, 0.150 mmol, 48.2% yield). LC-MS (ESI): m/z 476.3 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ 7.70 (s, 1H), 7.61 (d, J=8.3 Hz, 1H), 7.43 (d, J=3.2 Hz, 1H), 7.33 (s, 1H), 7.26 (t, J=7.9 Hz, 1H), 7.09 (d, J=7.4 Hz, 1H), 6.96 (s, 1H), 6.12 (d, J=3.0 Hz, 1H), 4.93 (s, 1H), 4.28 (t, J=5.5 Hz, 2H), 3.90 (s, 6H), 3.75 (t, J=5.5 Hz, 2H), 3.65-3.39 (m Hz, 4H), 1.68-1.40 (m, 6H).
To a solution of 2-(1H-indol-4-yl)-6,7-dimethoxy-4-(piperidine-1-carbonyl)isoquinolin-1(2H)-one (100 mg, 0.23 mmol) in DCM (15 mL) were added TEA (0.10 mL, 0.70 mmol), Ac2O (0.03 mL, 0.37 mmol) and DMAP (2.83 mg, 0.02 mmol). The resulting mixture was stirred at room temperature overnight. The mixture was purified by Prep-HPLC (system: Waters 2767/2545/2489/Qda, Waters sunfire C18 10 μm OBD 19*250 mm, Mobile Phase A: 0.1% FA in water, Mobile Phase B: CH3CN, Flow: 20 mL/min: Column temp: RT) to afford the title compound (45.0 mg, 0.10 mmol, 41.0% yield). LC-MS (ESI): m/z 474.2 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ 8.44 (d, J=8.2 Hz, 1H), 7.93 (d, J=3.8 Hz, l H), 7.70 (s, I H), 7.48 (t, J=8.0 Hz, 1H), 7.39 (s, 1H), 7.35 (d, J=7.6 Hz, 1H), 6.96 (s, l H), 6.46 (s, 1H), 3.90 (d, J=1.0 Hz, 6H), 3.64-3.40 (m, 4H), 2.69 (s, 3H), 1.64-1.45 (m, 6H).
To a solution of 2-(1H-indol-4-yl)-6,7-dimethoxy-4-(piperidine-1-carbonyl) isoquinolin-1(2H)-one (230 mg, 0.53 mmol) in DCM (20 mL) were added TEA (0.44 mL, 3.20 mmol), Cu(OAc)2 (194 mg, 1.07 mmol), cyclopropylboronic acid (180 mg, 2.12 mmol) and 4A molecular sieves (5.00 g). The resulting mixture was stirred at room temperature for 48 hr. The mixture was purified by Prep-HPLC (system: Waters 2767/2545/2489/Qda, Waters sunfire C18 10 μm OBD 19*250 mm, Mobile Phase A: 0.1% FA in water, Mobile Phase B: CH3CN, Flow: 20 mL/min: Column temp: RT) to afford the title compound (60.0 mg, 0.13 mmol, 23.9% yield). LC-MS (ESI): m/z 472.2 [M+H]+. 1H NMR (400 MHz, DMSO-d6) 7.68 (d, J=9.3 Hz, 2H), 7.40 (d, J=3.1 Hz, 1H), 7.36-7.28 (m, 2H), 7.13 (d, J=7.5 Hz, 1H), 6.96 (s, 1H), 6.09 (d, J=2.8 Hz, 1H), 3.89 (s, 6H), 3.57-3.44 (m, 4H), 1.64-1.43 (m, 6H), 1.37-1.19 (m, 1H), 1.14-1.06 (m, 2H), 1.03-0.95 (m, 2H).
To a mixture of ethyl 2-(2-ethoxy-2-oxoethyl)-4,5-dimethoxybenzoate (3.29 g, 11.1 mmol) and [(tert-butoxy)(dimethylamino)methyl]dimethylamine (3.10 g, 17.8 mmol) was stirred at 100° C. for 2 hr. The mixture was cooled to rt. AcOH (50 mL) and 6-fluoro-1H-indol-4-amine (2.0 g, 13.3 mmol) were added. Then the reaction mixture was stirred at 50° C. overnight. The mixture was diluted with H2O (100 mL) and the resulting mixture was extracted with EA (100 mL*2). The organic layers were combined, washed with brine (100 mL), dried over anhydrous Na2SO4, filtered and concentrated to afford ethyl 2-[(1Z)-3-ethoxy-1-[(6-fluoro-1H-indol-4-yl)amino]-3-oxoprop-1-en-2-yl]-4,5-dimethoxybenzoate (4.00 g, 8.76 mmol) as a yellow solid. LC-MS (ESI): m/z 457.3 [M+H]+
A mixture of ethyl 2-[(1Z)-3-ethoxy-1-[(6-fluoro-1H-indol-4-yl)amino]-3-oxoprop-1-en-2-yl]-4,5-dimethoxybenzoate (4.00 g, 8.76 mmol), LiOH·H2O (1.84 g, 43.8 mmol), THF (60 mL) and H2O (15 mL) was heated to 75° C., then the reaction was stirred at 75° C. for 5 hr. The mixture was adjusted to pH=2-3, then the mixture was concentrated under vacuum. The residue was filtered, the cake was washed with water(100 mL*2), dried to afford the crude product, which was slurried with EA and PE(100 mL, EA/PE=1/5) to afford 2-(6-fluoro-1H-indol-4-yl)-6,7-dimethoxy-1-oxo-1,2-dihydroisoquinoline-4-carboxylic acid (3.6 g, 9.42 mmol, 107.4%) as a brown solid. LC-MS (ESI): m/z 383.1 [M+H]+
To a solution of 2-(6-fluoro-1H-indol-4-yl)-6,7-dimethoxy-1-oxo-1,2-dihydroisoquinoline-4-carboxylic acid (3.50 g, 9.15 mmol) in DMF (50 mL) were added DIEA (4.55 mL, 27.5 mmol), HATU (4.18 g, 11.0 mmol) and the reaction was stirred at room temperature for 0.5 hr before piperidine (1.68 mL, 18.3 mmol) was added. Then the reaction mixture was stirred at room temperature for 1.5 hr. The mixture was diluted with H2O (20 mL) and the resulting mixture was extracted with EA (50 mL*2). The organic layers were combined, washed with brine (50 mL), dried over anhydrous Na2SO4, filtered and concentrated in vacuum. The residue was slurried with PE(80 mL) and EtOH (20 mL) to afford 2-(6-fluoro-1H-indol-4-yl)-6,7-dimethoxy-4-(piperidine-1-carbonyl)-1,2-dihydroisoquinolin-1-one (3.74 g, 8.33 mmol, 91% yield) as a grey solid. LC-MS (ESI): m/z 450.2 [M+H]+
To a solution of 2-(6-fluoro-1H-indol-4-yl)-4-(piperidine-1-carbonyl)-6,7-dimethoxy-1,2-dihydroisoquinolin-1-one (60.0 mg, 0.13 mmol) in DMF (5 mL) cooled to 0° C. was added NaH (4.62 mg, 0.19 mmol, 60% in oil), and the reaction was stirred at this temperature for 20 min. To the mixture was added CH3I (0.03 mL, 0.39 mmol), and the mixture was stirred at rt for 30 min. The reaction mixture was diluted with EA (20 mL) and brine (40 mL). The organic layer was separated, washed with further brine (40 mL), and concentrated in vacuo. The residue was purified by silica gel column chromatography eluting (PE/EA=10/1 to 1/2) to afford the title compound 2-(6-fluoro-1-methyl-1H-indol-4-yl)-4-(piperidine-1-carbonyl)-6,7-dimethoxy-1,2-dihydroisoquinolin-1-one (28 mg, 0.06 mmol, 45.3% yield) as a white solid.
The compounds below were synthesized following the procedures described for Compound 69.
1H NMR (400 MHz,
To a solution of 2-(6-fluoro-1H-indol-4-yl)-6,7-dimethoxy-1-oxo-1,2-dihydroisoquinoline-4-carboxylic acid (3.50 g, 9.15 mmol) in DMF (50 mL) were added DIEA (4.55 mL, 27.5 mmol), HATU (4.18 g, 11.0 mmol) and the reaction was stirred at room temperature for 0.5 hr before piperidine (1.68 mL, 18.3 mmol) was added. Then the reaction mixture was stirred at room temperature for 1.5 hr. The mixture was diluted with H2O (20 mL) and the resulting mixture was extracted with EA (50 mL*2). The organic layers were combined, washed with brine (50 mL), dried over anhydrous Na2SO4, filtered and concentrated in a vacuum. The residue was slurried with PE(80 mL) and EtOH (20 mL) to afford 2-(6-fluoro-1H-indol-4-yl)-6,7-dimethoxy-4-(piperidine-1-carbonyl)-1,2-dihydroisoquinolin-1-one (3.74 g, 8.33 mmol, 91% yield) as a grey solid. LC-MS (ESI): m/z 450.2 [M+H]+
A mixture of 2-(6-fluoro-1H-indol-4-yl)-6,7-dimethoxy-4-(piperidine-1-carbonyl)-1,2-dihydroisoquinolin-1-one (3.50 g, 7.79 mmol), cyclopropylboronic acid (2.68 g, 31.2 mmol), Cu(OAc)3 (5.66 g, 31.2 mmol), Na2CO3 (3.30 g, 31.3 mmol), 2-(pyridin-2-yl)pyridine (2.43 g, 15.6 mmol) and DCE (250 mL) was bubbled with argon for 15 seconds, then the reaction mixture was stirred at 70 C overnight. The mixture was diluted with DCM (80 mL), washed with brine (60 mL*2). The organic layer was separated, dried over anhydrous Na2SO4, filtered and concentrated. The residue was purified by silica gel column chromatography (EA/PE=1/10 to 3/1) to afford 2-(1-cyclopropyl-6-fluoro-1H-indol-4-yl)-6,7-dimethoxy-4-(piperidine-1-carbonyl)-1,2-dihydroisoquinolin-1-one (3.08 g, 6.26 mmol, 50.5% yield) as a white solid. LC-MS (ESI): m/z 490.2 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ 7.69 (s, 1H), 7.50 (dd, J=9.5, 1.6 Hz, 1H), 7.40 (d, J=3.3 Hz, 1H), 7.37 (s, 1H), 7.14 (dd, J=10.1, 2.1 Hz, 1H), 6.95 (s, 1H), 6.11 (d, J=3.1 Hz, 1H), 3.90 (s, 6H), 3.63-3.37 (m, 5H), 1.63-1.44 (m, 6H), 1.13-1.07 (m, 2H), 1.01-0.96 (m, 2H).
The compounds were synthesized following the procedures described for Compound 70
1H NMR (400 MHz, DMSO-d6) δ
To a solution of 2-bromo-5-methoxybenzoic acid (5.0 g, 21.64 mmol) in diethyl malonate (66.02 mL, 432.83 mmol) were added CuBr (621 mg, 4.33 mmol) and the reaction was stirred at 0° C., NaH (2.16 g, 54.1 mmol, 60% in oil) was added in portions. After the complete addition of sodium hydride, the mixture was stirred at 90° C. for 2 hours. The mixture was cooled to RT, water (200 mL) was added and the mixture was extracted with PE (200 mL*2). The water was combined, acidified with 6N HCl aq and the pH was adjusted to 2, then the mixture was extracted with EA (30 mL*2). The organic layers were combined, washed with brine (30 mL), dried over anhydrous Na2SO4, filtered and concentrated in a vacuum. The residue was purified by silica gel column chromatography (DCM/MeOH=500/1 to 50/1) to afford the titled compound (5.70 g, 18.37 mmol, 84.9% yield) as a green oil.
To a solution of 2-(1,3-diethoxy-1,3-dioxopropan-2-yl)-5-methoxybenzoic acid (5.70 g, 18.4 mmol) in EtOH (30 mL) were added Con HCl (35 mL, 351 mmol), then the mixture was stirred to 120° C. for 48 hours. The mixture was concentrated under a vacuum. The residue was carried water with anhydrous ethanol under a vacuum. The residue was added to 50 mL EtOH, then SOCl2 (10 mL) was added at 0˜-10° C., then the mixture was heated to reflux for 3 hours. The reaction was concentrated under vacuum to give a residue, which was purified by silica gel column chromatography eluting with (PE/EA=0/1 to 1/5) to afford the titled compound (2.10 g, 7.89 mmol, 42.9% yield) as a yellow oil. LC-MS (ESI): m/z 267.1 [M−18+H]+
A mixture of ethyl 2-(2-ethoxy-2-oxoethyl)-5-methoxybenzoate (300 mg, 1.13 mmol) and 1-tert-butoxy-N,N,N′N′-tetramethylmethanediamine (392.71 mg, 2.25 mmol) was stirred at 100° C. for 3 hr. The mixture was cooled to rt, then HOAc (8 mL) and 6-fluoro-1H-indol-4-amine (169.23 mg, 1.13 mmol) were added. Then the reaction mixture was stirred at 50° C. overnight. The mixture was concentrated in a vacuum. The residue was mixed with H2O (100 mL) and extracted with EA (100 mL*2). The organic layers were combined, washed with brine (100 mL), dried over anhydrous Na2SO4, filtered and concentrated. The residue was purified by silica gel column chromatography (PE/EA=10/1 to 4/1) to afford the titled compound (380 mg, 0.89 mmol, 79.1% yield) as a yellow oil. LC-MS (ESI): m/z 427.2 [M+H]+
To a solution of ethyl 2-(3-ethoxy-1-((6-fluoro-1H-indol-4-yl)amino)-3-oxoprop-1-en-2-yl)-5-methoxybenzoate (380 mg, 0.89 mmol) in THF (8 mL) and H2O (4 mL) was added LiOH·H2O (150 mg, 3.56 mmol) and the reaction mixture was stirred at 70° C. overnight. The mixture was acidified with 1 N HCl and the pH was adjusted to 2, a large amount of solids were extracted, then the mixture was filtered. The solid was collected, dried to afford the titled compound (200 mg, 0.57 mmol, 63.7% yield) as a gray solid. LC-MS (ESI): m/z 353.1 [M+H]+.
To a solution of 2-(6-fluoro-1H-indol-4-yl)-7-methoxy-1-oxo-1,2-dihydroisoquinoline-4-carboxylic acid (100 mg, 0.28 mmol) in DMF (4 mL) were added DIEA (0.09 mL, 0.57 mmol) and HATU (129 mg, 0.34 mmol). The reaction was stirred at room temperature for 30 min before piperidine (0.04 mL, 0.43 mmol) was added. The resulting reaction mixture was stirred at room temperature for 1.5 hr. The solution was quenched with water (20 mL) and a large amount of solid precipitation. The mixture was filtered. The solid was collected. The solid was mixed with EtOH (15 mL) and the mixture was stirred at 60° C. for 0.5 hr. The mixture was concentrated in vacuum. The residue was slurried with EtOH (4 mL) and PE (2 mL) to afford the titled compound (65.0 mg, 0.15 mmol, 54.6%). LC-MS (ESI): m/z 420.1 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ 11.48 (s, 1H), 7.75 (d, J=2.7 Hz, I H), 7.55 (d, J=8.9 Hz, 1H), 7.46 (dd, J=8.9, 2.8 Hz, I H), 7.43-7.40 (m, 1H), 7.37 (s, 1H), 7.34 (dd, J=9.6, 1.4 Hz, 1H), 7.09 (dd, J=10.2, 2.1 Hz, I H), 6.16 (s, 1H), 3.90 (s, 3H), 3.69-3.37 (m, 4H), 1.63-1.42 (m, 6H). 19F NMR (400 MHz, DMSO-d6) δ −121.81.
The compounds below were synthesized following the procedures described for Compound 71.
1H NMR (400 MHz, DMSO-d6) δ
To a suspension of 2-bromo-4-hydroxy-5-methoxybenzaldehyde (5.00 g, 21.6 mmol) in ACN (100 mL) was added Cs2CO3 (10.6 g, 32.5 mmol). The reaction mixture was stirred at room temperature for 15 min before CD3I (2.15 mL, 34.6 mmol) was added, then the reaction mixture was stirred at 50° C. for 18 hr. The mixture was cooled to ambient temperature, filtered through Celite and concentrated in vacuo. The residue was mixed with H2O (100 mL) and the resulting mixture was extracted with DCM (100 mL*2). The organic layers were combined, washed with brine (100 mL), dried over anhydrous Na2SO4, filtered and concentrated to afford the titled compound (5.50 g, 22.2 mmol, 100.0% yield) as a yellow solid. LC-MS (ESI): m/z 266.5 [M+18]+
To a solution of 2-bromo-5-methoxy-4-(methoxy-d3)benzaldehyde (5.5 g, 22.2 mmol) in H2O (210 mL) were added NaHCO3 (4.41 g, 52.541 mmol) and KMnO4 (11.0 g, 69.8 mmol). Then the reaction was stirred at 90° C. for 3 hr. The reaction was cooled to r.t, and the pH of this mixture was adjusted to 3˜4 with HCl (2M) and the resulting mixture was extracted with DCM (210 mL*2). The organic layers were combined, washed with brine (210 mL), dried over anhydrous Na2SO4, filtered and concentrated to afford the titled compound (3 g, 11.4 mmol, 51.2% yield) as a yellow solid. LC-MS (ESI): m/z 266.0 [M+H]+
To a solution of 2-bromo-5-methoxy-4-(methoxy-d3)benzoic acid (3.0 g, 11.4 mmol) in diethyl malonate (37.3 g, 233 mmol) was added CuBr (0.33 g, 2.27 mmol) and the reaction was stirred at 0° C., NaH (1.18 g, 29.5 mmol, 60% in oil) was added in portions. After the complete addition of sodium hydride, the mixture was heated to 90° C. for 2 hours. The mixture was cooled to RT, water (200 mL) was added and the resulting mixture was extracted with PE (200 mL*2). The water was combined, acidified with 6N HCl aq and the pH was adjusted to 2, a large amount of solids were released. Then the mixture was filtered. The solid was collected, dried to afford the title compound the titled compound (4.00 g, 11.7 mmol, 100.0% yield) as a yellow solid. LC-MS (ESI): m/z 326.1 [M−18+H]+
To a solution of 2-(1,3-diethoxy-1,3-dioxopropan-2-yl)-5-methoxy-4-(methoxy-d3)benzoic acid (4.00 g, 11.7 mmol) in EtOH (30 mL) were added Con·HCl (30 mL, 300 mmol), then the mixture was stirred to 120° C. for 48 hours. The mixture was concentrated under a vacuum. The residue was carried water with anhydrous ethanol under a vacuum. The residue was added to 50 mL EtOH, then SOCl2 (10 mL) was added at 0˜10° C., then the mixture was heated to reflux for 3 hours. The reaction was concentrated under vacuum to give a residue, which was purified using silica gel column chromatography (EA/PE=0/1 to 1/5) to afford the title compound the titled compound (2.50 g, 8.35 mmol, 71.7% yield) as a white solid. LC-MS (ESI): m/z 326.1 [M−18+H]+
A mixture of ethyl 2-(2-ethoxy-2-oxoethyl)-5-methoxy-4-(methoxy-d3)benzoate (679 mg, 2.27 mmol) and 1-tert-butoxy-N,N,N′,N′-tetramethylmethanediamine (791.21 mg, 4.540 mmol) was stirred at 100° C. for 2 hr. The mixture was cooled to rt, then HOAc (15 mL) and 1H-indol-4-amine (300 mg, 2.27 mmol) were added. The reaction mixture was stirred at 50° C. overnight. The reaction mixture was concentrated in a vacuum. The reduise was mixed with EA (100 mL), then washed with H2O (100 mL). The organic layers were combined, washed with brine (100 mL), dried over anhydrous Na2SO4, filtered and concentrated. The residue was purified by silica gel column chromatography (EA/PE=1/20 to 1/3) to afford the titled compound (500 mg, 1.13 mmol, 50.0% yield) as a yellow solid. LC-MS (ESI): m/z 442.1 [M+H]+
To a solution of ethyl 2-(1-((1H-indol-4-yl)amino)-3-ethoxy-3-oxoprop-1-en-2-yl)-5-methoxy-4-(methoxy-d3)benzoate (500 mg, 1.13 mmol) in THF (10 mL) and H2O (5 mL) was added LiOH·H2O (190 mg, 4.53 mmol) and the reaction mixture was stirred at 80° C. overnight, then cooled to rt, adjusted to pH=2-3 with con. HCl. The mixture was extracted with EA (30 mL*2). The organic layers were combined, washed with brine (50 mL), dried over anhydrous Na2SO4, filtered and concentrated in a vacuum to afford the title compound the titled compound (300 mg, 0.82 mmol, 72.1% yield) as a yellow solid. LC-MS (ESI): m/z 368.0 [M+H]+
To a solution of 2-(1H-indol-4-yl)-7-methoxy-6-(methoxy-d3)-1-oxo-1,2-dihydroisoquinoline-4-carboxylic acid (300 mg, 0.817 mmol) in DMF (6 mL) were added DIEA (0.41 mL, 2.450 mmol), HATU (373 mg, 0.980 mmol) and piperidine (0.09 mL, 0.980 mmol). Then the reaction mixture was stirred at room temperature for 2 hr. The mixture was mixed with H2O (100 mL) and the resulting mixture was extracted with EA (50 mL*3). The organic layers were combined, washed with brine (100 mL), dried over anhydrous Na2SO4, filtered and concentrated. The residue was purified by silica gel column chromatography (DCM/MeOH=1/50 to 1/30) to afford the titled compound (150 mg, 0.35 mmol, 42.3% yield). LC-MS (ESI): m/z 435.3 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ 11.41 (s, 1H), 7.71 (s, 1H), 7.52 (d, J=8.1 Hz, 1H), 7.41 (t, J=2.6 Hz, 1H), 7.35 (s, 1H), 7.22 (t, J=7.8 Hz, 1H), 7.07 (d, J=7.4 Hz, 1H), 6.96 (s, 1H), 6.13 (s, 1H), 3.90 (s, 3H), 3.51 (s, 4H), 1.66-1.40 (m, 6H).
A mixture of 1,4-dithiane-2,5-diol (10 g, 65.68 mmol), dimethyl 3-oxopentanedioate (33.93 g, 197 mmol) and LiBr (1.17 g, 19.7 mmol) in dioxane (300 mL) was heated to reflux for 12 hr and cooled to RT. The whole was filtered in a vacuum and the filter cake was washed with Et2O (50 mL). Then the filtrate was concentrated in a vacuum. The residue was purified by column chromatography (PE/EA=100/1-15/1) to give the title product (14 g, 65.35 mmol, 99.5% yield). LCMS: calc. for [C9H11O4S]+:215.03, found: 215.0 [M+H]+
A mixture of methyl 2-(2-methoxy-2-oxoethyl)thiophene-3-carboxylate (3.0 g, 14.0 mmol) and 1-tert-butoxy-N,N,N′,N′-tetramethylmethanediamine (5.81 mL, 28.0 mmol) was stirred at 100° C. for 3.0 h. After cooling to rt, the resulting mixture we treated with HOAc (30 mL) and 5-methylbenzo[d]isoxazol-3-amine (6.22 g, 42.0 mmol). The resulting mixture was stirred at room temperature for 16 hr. Then the reaction mixture was diluted with water (10 mL) and the whole was extracted with EtOAc (30 mL×3). The organic layers were combined, washed with brine (30 mL×3), dried over anhydrous Na2SO4, filtered. The filtrate was concentrated under reduced pressure. The residue was purified using silica gel column chromatography (PE/EA=15/1) to afford the title compound (200 mg, 0.59 mmol, 4.2% yield). LCMS: calc. for [C17H13N2O4S]+:341.05, found: 341.0 [M+H]+
To a solution of methyl 5-(5-methylbenzo[d]isoxazol-3-yl)-4-oxo-4,5-dihydrothieno[3,2-c]pyridine-7-carboxylate (200 mg, 0.588 mmol) in MeOH (15 mL) and H2O (15 mL) was added LiOH·H2O (112.6 mg, 2.68 mmol) and the resulting mixture was heated to 70 C for 2 hours. The reaction mixture was cooled to rt and adjusted to pH 8-9 with diluted HCl (1.0 M). The resulting mixture was concentrated under reduced pressure to remove most of MeOH. The residue was adjusted to pH 2-3 with diluted HCl (1.0 M) and the whole was extracted with EtOAc (50 mL×2). The organic layers were combined, dried over anhydrous Na2SO4 and filtered. The filtrate was concentrated under reduced pressure to afford the titled compound (124 mg, 0.38 mmol, 35.9% yield). LCMS: calc. for [C16H10N2O4S]+: 327.04, found: 326.9 [M+H]+
A mixture of 5-(5-methylbenzo[d]isoxazol-3-yl)-4-oxo-4,5-dihydrothieno[3,2-c]pyridine-7-carboxylic acid (124 mg, 0.349 mmol), piperidine (0.14 mL, 1.397 mmol), HATU (159 mg, 0.419 mmol), DIEA (0.17 mL, 1.05 mmol) and DMF (5.0 mL) was stirred at RT for 2 h. The reaction mixture was mixed with brine (10 mL) and EA (10 mL). The organic layer was separated, and concentrated under reduced pressure. The residue was purified by pre-HPLC (Base method: aqueous ammonia in MeCN) to afford the titled compound (18.96 mg, 0.04 mmol, 12.6% yield). LCMS: calc. for [C21H20N3O3S]+:394.11, found: 394.0 [M+H]+. 1H NMR (400 MHz, DMSO) δ 7.93 (s, 1H), 7.87 (d, J=5.4 Hz, 1H), 7.78 (d, J=8.6 Hz, 1H), 7.66 (d, J=5.3 Hz, 1H), 7.62-7.52 (m, 2H), 2.43 (s, 3H), 3.56-3.68 (m, 4H), 1.62 (d, J=4.4 Hz, 2H), 1.53 (s, 4H).
A mixture of 2-bromo-5-hydroxy-4-methoxybenzoic acid (5 g, 20.24 mmol), H2SO4 (0.27 mL, 5.06 mmol) in EtOH (50 mL) was stirred at 80° C. overnight. The mixture was diluted with H2O (100 mL) and the resulting mixture was extracted with DCM (100 mL*2). The organic layers were combined, washed with brine (100 mL), dried over anhydrous Na2SO4, filtered and concentrated to afford the titled compound (5.3 g, 19.27 mmol, 95.2% yield) as a white solid. LC-MS (ESI): m/z 273.1/275.1 [M−H]−
A mixture of ethyl 2-bromo-5-hydroxy-4-methoxybenzoate (1.0 g, 3.64 mmol), sodium 2-chloro-2,2-difluoroacetate (1.39 g, 9.09 mmol), Cs2CO3 (2.37 g, 7.27 mmol) in DMF (3 mL) was stirred at 80° C. for 3 hr. The mixture was diluted with H2O (100 mL) and the resulting mixture was extracted with DCM (100 mL*2). The organic layers were combined, washed with brine (100 mL), dried over anhydrous Na2SO4, filtered and concentrated. The residue was purified by silica gel column chromatography (EA/PE=1/50-1/20) to afford the titled compound (780 mg, 2.40 mmol, 66.0% yield) as a white solid. LC-MS (ESI): m/z 325.0/327.0 [M+H]+
To a solution of ethyl 2-bromo-5-(difluoromethoxy)-4-methoxybenzoate (1.0 g, 3.08 mmol) in 1,3-diethyl propanedioate (10.10 g, 63.06 mmol) were added CuBr (0.09 g, 0.62 mmol), and the reaction was stirred at 0° C., NaH (0.19 g, 8.0 mmol, 60% in oil) was added in portions. After the complete addition of sodium hydride, the mixture was stirred at 90° C. for 2 hours. The mixture was cooled to RT, water (200 mL) was added and the mixture was extracted with EA (100 mL×2). The organic layers were combined, washed with brine (200 mL), dried over anhydrous Na2SO4, filtered and concentrated in a vacuum. The residue was purified by silica gel column chromatography (EA/PE=1/50-1/20) to afford the titled compound (1.1 g, 2.72 mmol, 88.4% yield) as a yellow liquid. LC-MS (ESI): m/z 405.1 [M+H]
To a solution of 2-(1,3-diethoxy-1,3-dioxopropan-2-yl)-4-({circumflex over ( )}H3)methoxy-5-methoxybenzoic acid (4.0 g, 11.65 mmol) in EtOH (30 mL) were added Con HCl (30 mL, 300 mmol), then the mixture was heated to 120° C. and stirred at this temperature for 24 hours. The reaction was concentrated under vacuum to give a residue, which was purified using silica gel column chromatography eluting with ethyl acetate in petroleum ether (EA/PE=0/1-1/1) afford the titled compound (300 mg, 1.06 mmol, 39.1% yield) as a white solid. LC-MS (ESI): m/z 283.1 [M+H]+
A mixture ethyl 2-(2-ethoxy-2-oxoethyl)-5-hydroxy-4-methoxybenzoate (300 mg, 1.06 mmol), sodium 2-chloro-2,2-difluoroacetate (405.1 mg, 2.66 mmol) and Cs2CO3 (692.52 mg, 2.13 mmol) in DMF (10 mL). Then the reaction mixture was stirred at 80° C. for 3 hr. The mixture was diluted with H2O (200 mL) and the resulting mixture was extracted with EA (100 mL*3). The organic layers were combined, washed with brine (200 mL*3), dried over anhydrous Na2SO4, filtered and concentrated to afford the titled compound (90 mg, 0.27 mmol, 25.5% yield) as a yellow solid. LC-MS (ESI): m/z 333.1 [M+H]+
To a mixture of ethyl 5-(difluoromethoxy)-2-(2-ethoxy-2-oxoethyl)-4-methoxybenzoate (90 mg, 0.27 mmol) and [(teRT-butoxy)(dimethylamino)methyl]dimethylamine (94.4 mg, 0.54 mmol) was stirred at 100° C. for 2 hr. The mixture was cooled to RT. CH3COOH (10 mL) and 6-fluoro-1H-indol-4-amine (61.00 mg, 0.41 mmol) were added. Then the reaction mixture was stirred at 50° C. overnight. The mixture was diluted with H2O (100 mL) and the resulting mixture was extracted with EA (100 mL*2). The organic layers were combined, washed with brine (100 mL), dried over anhydrous Na2SO4, filtered and concentrated. The residue was purified by silica gel column chromatography (EA/PE=1/10-1/5) to afford the titled compound (100 mg, 0.20 mmol, 75.0% yield) as a yellow solid. LC-MS (ESI): m/z 493.2 [M+H]+
To a solution of ethyl 5-(difluoromethoxy)-2-[(1Z)-3-ethoxy-1-[(6-fluoro-1H-indol-4-yl)amino]-3-oxoprop-1-en-2-yl]-4-methoxybenzoate (100 mg, 0.20 mmol) in THF (6 mL) and H2O (3 mL) was added LiOH·H2O (34.1 mg, 0.81 mmol) and the reaction mixture was stirred at 80° C. for 18 hr. The mixture was acidified with 6 N HCl and the pH was adjusted to 2, then the mixture was filtered. The solid was collected and dried to afford the titled compound (115 mg, 0.19 mmol, 95.9% yield) as a black solid. LC-MS (ESI): m/z 419.1[M+H]+
To a solution of 7-(difluoromethoxy)-2-(6-fluoro-1H-indol-4-yl)-6-methoxy-1-oxo-1,2-dihydroisoquinoline-4-carboxylic acid (125 mg, 0.30 mmol) in DMF (2 mL) were added DIEA (0.15 mL, 0.90 mmol), HATU (170.4 mg, 0.45 mmol) and piperidine (0.05 mL, 0.60 mmol). Then the reaction mixture was stirred at room temperature for 2.5 hr. The mixture was diluted with H2O (50 mL) and the resulting mixture was extracted with DCM (50 mL*2). The organic layers were combined, washed with brine (100 mL), dried over anhydrous Na2SO4, filtered and concentrated. The residue was purified by silica gel column chromatography (EA/PE=1/10-1/1) to afford the titled compound(60 mg, 0.12 mmol, 41.3% yield) as a light brown solid. LC-MS (ESI): m/z 486.3 [M+H]+. 1H NMR (400 MHz, DMSO-d6)) δ 11.49 (s, 1H), 7.99 (s, 1H), 7.53 (s, 1H), 7.50 (s, 0.2H), 7.44-7.40 (m, 1H), 7.34 (dd, J=9.5, 1.4 Hz, 1H), 7.31 (s, 0.5H), 7.14-7.07 (m, 2.3H), 6.17 (s, 1H), 3.96 (s, 3H), 3.65-3.44 (m, 4H), 1.64-1.46 (m, 6H). 19F NMR(400 MHz, DMSO-d6) δ −82.18, -121.81/121.82.
To a solution of 7-(difluoromethoxy)-2-(6-fluoro-1H-indol-4-yl)-6-methoxy-4-(piperidine-1-carbonyl)-1,2-dihydroisoquinolin-1-one (30 mg, 0.06 mmol) in THF (6 mL) was added NaH (4.94 mg, 0.12 mmol, 60% in oil) under 0° C. The reaction was stirred at this temperature for 30 min before CH3I (17.54 mg, 0.12 mmol) was added. The mixture was stirred at RT for 1 hr. The mixture was diluted with EA (50 mL), washed with brine (50 mL). The organic layer was separated, dried over anhydrous Na2SO4, filtered and concentrated. The residue was purified by silica gel column chromatography (EA/PE=1/10-1/5) to afford the titled compound (10 mg, 0.02 mmol, 32.4% yield). LC-MS (ESI): m/z 500.2 [M+H]+. 1H NMR (400 MHz, DMSO-d6)) δ 7.99 (s, 1H), 7.53-7.49 (m, 2.3H), 7.41 (d, J=3.2 Hz, 1H), 7.31 (s, 0.4H), 7.15-7.11 (m, 1.4H), 7.10 (s, 1H), 6.16 (d, J=2.9 Hz, 1H), 3.96 (s, 3H), 3.83 (s, 3H), 3.59-3.45 (m, 4H), 1.62-1.46 (m, 6H). 19F NMR(400 MHz, DMSO-d6) δ −82.20, -121.03/121.04.
The compounds below were synthesized following the procedures described for Compound 78.
1H NMR (400
A mixture of 6,7-dimethoxy-2-(5-methylbenzo[d]isoxazol-3-yl)-1-oxo-1,2-dihydroisoquinoline-4-carboxylic acid (20.0 mg, 0.053 mmol), Cu (0.67 mg, 0.011 mmol) and quinolone (3.0 mL) was stirred at 200° C. for 1.0 hr. The mixture was purified by Pre-HPLC (Waters 2767/2545/2489, Waters Xbridge C18 10 μm OBD 19*250 mm, Mobile Phase A: 0.1% NH4OH in water, Mobile Phase B: CH3CN, Flow: 20 mL/min, Column temp: RT) to afford the title compound (1.72 mg, 0.01 mmol, 1.9% yield). LC-MS(ESI):m/z 337.2[M+H]+. 1H NMR (400 MHz, DMSO-d6): δ 7.77 (d, J=8.7 Hz, 1H), 7.68 (s, 1H), 7.59-7.56 (m, 1H), 7.54-7.52 (m, 2H), 7.34 (s, 1H), 6.79 (d, J=7.5 Hz, 1H), 3.95 (s, 3H), 3.91 (s, 3H), 2.43 (s, 3H).
To a solution of 6,7-dimethoxy-2-(5-methylbenzo[d]isoxazol-3-yl)-1-oxo-1,2-dihydroisoquinoline-4-carboxylic acid (1.50 g, 3.94 mmol) in DMF (20 mL) were added DIEA (5.23 mL, 31.6 mmol), EDCI (1.13 g, 5.92 mmol) and HOBt (799 mg, 5.92 mmol), then the reaction was stirred at 50° C. for 8 hrs. The resulting mixture was mixed with EA (100 mL) and brine (120 mL). The organic layer was separated and the water phase was extracted with EA (50 mL). Then the organic layers were combined and washed with brine (50 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified using silica gel column chromatography (EA/PE=1/100-1/1) to afford the title compound (417 mg, 0.98 mmol, 25.0% yield) as a yellow solid. LCMS: calc. for [C22H22N3O3]+: 424.14, found: 424.1 [M+H]+
To a solution of N,6,7-trimethoxy-N-methyl-2-(5-methylbenzo[d]isoxazol-3-yl)-1-oxo-1,2-dihydroisoquinoline-4-carboxamide (50 mg, 0.12 mmol) in THE (40 mL) was added phenyllithium (0.24 mL, 0.24 mmol, 1 M in ether) drop wise at −30° C., and the reaction was stirred at −30-−10° C. for 1.0 hour. Then the reaction mixture was diluted with EA (30 mL) and brine (20 mL). The organic layer was separated, dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by Pre-HPLC (Method: Waters 2767/2545/2489, Inertsil ODS-3, 10 μm 20*250 nm, Mobile Phase A: 0.1% FA in water, Mobile Phase B: CH3CN, Flow: 20 mL/min, Column temp: RT) to afford the title compound (9.32 mg, 0.02 mmol, 15.7% yield). LCMS: calc. for [C28H28N3O3]+: 502.19, found: 502.1[M+H]+. 1H NMR (400 MHz, DMSO) δ 7.59 (t, J=6.5 Hz, 3H), 7.48 (d, J=8.6 Hz, 1H), 7.29 7.34 (m, 2H), 7.21-7.28 (m, 3H), 6.93 (s, 1H), 5.75 (s, 1H), 4.61 (s, 1H), 3.98 (s, 3H), 3.86 (s, 3H), 3.77 (s, 3H), 3.12 (s, 3H), 2.43 (s, 3H).
To a solution of N,6,7-trimethoxy-N-methyl-2-(5-methylbenzo[d]isoxazol-3-yl)-1-oxo-1,2-dihydroisoquinoline-4-carboxamide (350 mg, 0.83 mmol) in THE (60 mL) was added DIBAL-H (1.10 mL, 1.65 mmol, 1.5 M in toluene) drop wise at −30° C. and the reaction mixture was stirred at −30-−10° C. for 2 hrs. The resulting mixture was mixed with EA (60 mL) and brine (30 mL). The organic layer was separated, dried over anhydrous Na2SO4, filtered in a vacuum. The filtrate was concentrated under reduced pressure. The residue was purified using silica gel column chromatography eluting with methanol in DCM (0˜10%) to afford the title compound (171 mg, 0.36 mmol, 43.5% yield) as a yellow solid. LCMS: calc. for [C2MH17FN2O5]+: 365.11, found: 365.0 [M+H]+
To a suspension of 6,7-dimethoxy-2-(5-methylbenzo[d]isoxazol-3-yl)-1-oxo-1,2-dihydroisoquinoline-4-carbaldehyde (50 mg, 0.14 mmol) in THF (20 mL) and MeOH (20 mL) was added NaBH4 (5.0 mg, 0.14 mmol) at RT, then the reaction mixture became clear and the mixture was stirred at RT for 1 hr before con. HCl (0.20 mL) was added. Then the resulting mixture was concentrated under reduced pressure. The residue was purified by pre-HPLC (Method: Waters 2767/2545/2489, Inertsil ODS-3 10 μm 20*250 nm, Mobile Phase A: 0.1% FA in water, Mobile Phase B: CH3CN, Flow: 20 mL/min, Column temp: RT) to afford the title compound (9.0 mg, 0.02 mmol, 16.8% yield). LCMS: calc. for [C20H19FN2O5]+: 367.12, found: 367.0 [M+H]+. 1H NMR (400 MHz, DMSO) δ 7.77 (d, J=8.7 Hz, 1H), 7.73 (s, 1H), 7.58 (d, J=8.6 Hz, 1H), 7.54 (s, 1H), 7.51 (s, 1H), 7.32 (s, 1H), 5.31 (t, J=5.2 Hz, 1H), 4.65 (d, J=4.8 Hz, 2H), 3.97 (s, 3H), 3.92 (s, 3H), 2.43 (s, 3H).
To a solution of 6,7-dimethoxy-2-(5-methylbenzo[d]isoxazol-3-yl)-1-oxo-1,2-dihydroisoquinoline-4-carbaldehyde (70 mg, 0.19 mmol) in THE (20 mL) was added phenyllithium (65 mg, 0.77 mmol, 1.0 M in ether) at 0° C. The reaction mixture was warmed to RT and stirred for 2 hr. The resulting mixture was mixed with EA (10 mL) and brine (10 mL). The organic layer was separated, dried over anhydrous Na2SO4, filtered in vacuum. The filtrate was concentrated under reduced pressure. The residue was purified by pre-HPLC (Method: Waters 2767/2545/2489, Inertsil ODS-3 10 μm 20*250 nm, Mobile Phase A: 0.10% FA in water, Mobile Phase B: CH3CN, Flow: 20 mL/min, Column temp: RT) to afford 13 mg of crude product, which was purified by Chrial-HPLC (OJ-70(Flow: 70 g/min)-30(30% MeOH)/70(70% CO2)+0.1% NH3·H2O) to afford the title compound (3.79 mg, 0.01 mmol, 4.5% yield) as a white solid. LCMS: calc. for [C25H23N2O5]: 443.15, found: 443.1 [M+H]+. 1H NMR (400 MHz, DMSO) δ 7.78 (d, J=9.4 Hz, 1H), 7.70 (s, 1H), 7.56-7.63 (m, 3H), 7.34 (t, J=7.5 Hz, 2H), 7.24 (s, 2H), 7.52 (d, J=7.6 Hz, 2H), 6.16 (d, J=4.2 Hz, 1H), 3.87 (s, 3H), 6.01 (s, 1H), 3.73 (s, 3H), 2.44 (s, 3H).
To a solution of 4-(hydroxy(phenyl)methyl)-6,7-dimethoxy-2-(5-methylbenzo[d]isoxazol-3-yl)isoquinolin-l(2H)-one (95 mg, 0.22 mmol) in DCM (15 mL) was added MnO2 (500 mg, 5.75 mmol, 80% of purity) at 40° C. and the reaction was stirred at this temperature for 6 hr. The mixture was filtered in vacuum. The filtrate was concentrated under reduced pressure. The residue was purified by Pre-HPLC (Base method: NH3—H2O in CH3CN) to afford the title compound (14.65 mg, 0.03 mmol, 15.5% yield). LCMS: calc. for [C26H20N2O5]+:440.14, found: 441.1 [M+H]+. 1H NMR (400 MHz, DMSO) δ 7.88 (dd, J=9.4, 4.7 Hz, 3H), 7.80 (s, 1H), 7.75 (d, J=8.7 Hz, 1H), 7.67 (s, 11H), 7.61 (s, 1H), 7.57 (d, J=7.7 Hz, 2H), 3.96 (s, 2H), 3.90 (s, 2H), 2.44 (s, 2H).
The compounds below were synthesized following the procedures described for Compound 101 and 102.
1H NMR (400 MHz, DMSO-d6) δ
To a solution of ethyl 2-(2-ethoxy-2-oxoethyl)-4,5-dihydroxybenzoate (1 g, 3.73 mmol) in DMF (10 mL) was added NaH (0.20 g, 8.20 mmol, 60% in oil) under N2 atmosphere. The resulting mixture was stirred at 0° C. for 0.5 hr before fluoro-iodo-methane (1.31 g, 8.201 mmol) was added. The reaction mixture was stirred at rt for 2 hr under an N2 atmosphere. The resulting mixture was quenched with H2O (10 mL) and extracted with EA (30 mL*2). The organic layers were combined, washed with brine (30 mL), dried over anhydrous Na2SO4, filtered and concentrated. The residue was purified by silica gel column chromatography (PE/EA=50/1 to 1/1) to afford ethyl 2-(2-ethoxy-2-oxoethyl)-4,5-bis(fluoromethoxy)benzoate (800 mg, 2.41 mmol, 64.6% yield) as a white solid. LC-MS (ESI): m/z 334.2 [M+H]+.
To a mixture of ethyl 2-(2-ethoxy-2-oxoethyl)-4,5-bis(fluoromethoxy)benzoate (500 mg, 1.51 mmol) and [(tert-butoxy)(dimethylamino)methyl]dimethylamine (393.4 mg, 2.26 mmol) was stirred at 100° C. for 2 hr. The mixture was cooled to rt. AcOH (10 mL) and 6-fluoro-1H-indol-4-amine (248.5 mg, 1.66 mmol) were added. Then the reaction mixture was stirred at 50° C. overnight. The mixture was diluted with H2O (600 mL) and a large amount of solid was precipitated. The mixture was filtered to afford ethyl 2-[(1Z)-3-ethoxy-1-[(6-fluoro-1H-indol-4-yl)amino]-3-oxoprop-1-en-2-yl]-4,5-bis(fluoromethoxy)benzoate (484 mg, 0.98 mmol, 65.3% yield) as a brown solid. LC-MS (ESI): m/z 491.2 [M−H]−.
To a solution of ethyl 2-[(1Z)-3-ethoxy-1-[(6-fluoro-1H-indol-4-yl)amino]-3-oxoprop-1-en-2-yl]-4,5-bis(fluoromethoxy)benzoate (484 mg, 0.98 mmol) in THE (14 mL) and H2O (7 mL) was added LiOH·H2O (206.2 mg, 4.91 mmol) and the reaction mixture was stirred at 50° C. overnight. THE was removed in a vacuum. The resulting mixture was acidified with 2N HCl and the pH was adjusted to 1˜2, a large amount of solid was precipitated. Then the mixture was filtered. The solid was collected, dried to afford 2-(6-fluoro-1H-indol-4-yl)-6,7-bis(fluoromethoxy)-1-oxo-1,2-dihydroisoquinoline-4-carboxylic acid (536 mg, 1.28 mmol, 130.3% yield) as a brown solid. LC-MS (ESI): m/z 419.1 [M+H]−.
To a solution of 2-(6-fluoro-1H-indol-4-yl)-6,7-bis(fluoromethoxy)-1-oxo-1,2-dihydroisoquinoline-4-carboxylic acid (200 mg, 0.49 mmol) in DMF (10 mL) were added DIEA (0.40 mL, 2.39 mmol), HATU (218.1 mg, 0.57 mmol), and 4-fluoropiperidine (59.2 mg, 0.57 mmol). Then the reaction mixture was stirred at room temperature for 2 hr. The mixture was diluted with H2O (50 mL) and the resulting mixture was extracted with EA (50 mL*2). The organic layers were combined, washed with brine (100 mL), dried over anhydrous Na2SO4, filtered and concentrated. The residue was purified by silica gel column chromatography (EA/PE=1/10 to 2/1) to afford 2-(6-fluoro-1H-indol-4-yl)-6,7-bis(fluoromethoxy)-4-(4-fluoropiperidine-I-carbonyl)-1,2-dihydroisoquinolin-1-one (120 mg, 0.24 mmol, 49.9% yield) as white solid. LC-MS (ESI): m/z 502.2 [M−H]−. 1H NMR (400 MHz, DMSO-d6) δ 11.49 (s, 1H), 8.06 (s, 1H), 7.61 (s, 1H), 7.44-7.41 (m, 1H), 7.37-7.33 (m, 2H), 7.37-7.33 (m, 2H), 6.19 (s, 1H), 6.08 (s, 2H), 5.95 (s, 2H), 4.97-4.79 (m, 1H), 3.68-3.48 (m, 4H), 1.95-1.67 (m, 4H).
To a solution of 2-(6-fluoro-1H-indol-4-yl)-6,7-bis(fluoromethoxy)-4-(4-fluoropiperidine-1-carbonyl)-1,2-dihydroisoquinolin-1-one (50 mg, 0.10 mmol) in DMF (5 mL) was added NaH (2.86 mg, 0.12 mmol, 60% in oil) under 0° C. The reaction was stirred at this temperature for 30 min before CH3I (0.01 mL, 0.12 mmol) was added. The mixture was stirred at rt for 2 hr. The mixture was diluted with H2O (50 mL) and the resulting mixture was extracted with EA (50×3). The organic layer was combined, washed with brine (50 mL), dried over anhydrous Na2SO4, filtered and concentrated in vacuum. The residue was purified by flash chromatography (PE/EA=200/1 to 0/1) to afford 2-(6-fluoro-1-methyl-1H-indol-4-yl)-6,7-bis(fluoromethoxy)-4-(4-fluoropiperidine-1-carbonyl)-1,2-dihydroisoquinolin-1-one (28.01 mg, 0.05 mmol, 54.5% yield) as a white solid. LC-MS (ESI): m/z 518.2 [M+H]−. 1H NMR (400 MHz, DMSO-d6) δ 8.05 (s, 1H), 7.59 (s, 1H), 7.52 (dd, J=9.7, 1.8 Hz, 1H), 7.41 (d, J=3.1 Hz, 1H), 7.34 (s, 1H), 7.14 (dd, J=10.0, 1.7 Hz, 1H), 6.19 (d, J=3.0 Hz, 1H), 6.08 (s, 2H), 5.95 (s, 2H), 4.97-4.81 (m, 1H), 3.83 (s, 3H), 3.69-3.53 (m, 4H), 1.94-1.82 (m, 2H), 1.77-1.67 (m, 2H).
The compounds below were synthesized following the procedures described for Compound 157.
1H NMR (400 MHz, DMSO-d6) δ
To a solution of ethyl 2-(2-ethoxy-2-oxoethyl)-4,5-dimethoxybenzoate (10 g, 33.7 mmol) in DCM (300 mL) under −78° C. was added BBr3 in DCM (337 mL, 337 mmol). The reaction was stirred at this temperature for an hour, then the mixture was stirred at rt overnight. The mixture was concentrated and dried to afford a crude mixture of 2-(carboxymethyl)-4,5-dihydroxybenzoic acid, 2-(2-ethoxy-2-oxoethyl)-4,5-dihydroxybenzoic acid and 2-[2-(ethoxycarbonyl)-4,5-dihydroxyphenyl]acetic acid (8.0 g, 33.3 mmol, 98.7% yield) as a yellow solid. LC-MS (ESI): m/z 241.1 [M+H]+.
To a solution of 2-(carboxymethyl)-4,5-dihydroxybenzoic acid, 2-(2-ethoxy-2-oxoethyl)-4,5-dihydroxybenzoic acid and 2-[2-(ethoxycarbonyl)-4,5-dihydroxyphenyl]acetic acid (8 g, 33.3 mmol) in EtOH (150 mL) was added H2SO4 (5 mL). Then the reaction mixture was stirred at 80° C. overnight. The mixture was adjusted to pH=4-5 with 2M NaOH solution, a large amount of solid was precipitated. Then the mixture was filtered. The solid was collected and slurried with EA and PE (300 mL, EA/PE=1/6) to afford ethyl 2-(2-ethoxy-2-oxoethyl)-4,5-dihydroxybenzoate (7 g, 26.09 mmol, 78.4% yield) as a yellow solid. LC-MS (ESI): m/z 269.1 [M+H]+.
To a solution of ethyl 2-(2-ethoxy-2-oxoethyl)-4,5-dihydroxybenzoate (1 g, 3.73 mmol) in DMF (10 mL) was added NaH (196.8 mg, 8.20 mmol, 60% in oil) under N2 atmosphere. The resulting mixture was stirred at this temperature for 0.5 hour before CD3I (1.19 g, 8.20 mmol) was added. The reaction mixture was stirred at rt for 2 hours under an N2 atmosphere. The resulting mixture was quenched with H2O (10 mL) and extracted with EA (30 mL*2). Organic layers were combined, washed with brine (30 mL), dried over anhydrous Na2SO4, filtered and concentrated. The residue was purified by silica gel column chromatography (PE/EA=50/1 to 1/1) to afford ethyl 2-(2-ethoxy-2-oxoethyl)-4,5-bis(methoxy-d3)benzoate(1.03 g, 3.39 mmol, 91.0% yield) as a white solid. LC-MS (ESI): m/z 303.3 [M+H]+.
To a mixture of ethyl 2-(2-ethoxy-2-oxoethyl)-4,5-bis(methoxy-d3)benzoate (300 mg, 1.0 mmol) and [(tert-butoxy)(dimethylamino)methyl]dimethylamine (259.3 mg, 1.49 mmol) was stirred at 100° C. for 2 hours. The mixture was cooled to RT. AcOH (10 mL) and 6-fluoro-1H-indol-4-amine (163.9 mg, 1.09 mmol) were added. Then the reaction mixture was stirred at 50° C. overnight. The mixture was diluted with H2O (600 mL) and a large amount of solid was precipitated. The mixture was filtered to afford ethyl 2-(3-ethoxy-1-((6-fluoro-1H-indol-4-yl)amino)-3-oxoprop-1-en-2-yl)-4,5-bis(methoxy-d3)benzoate (434 mg, 0.94 mmol, 94.6% yield) as a brown solid. LC-MS (ESI): m/z 463.3[M+H]+.
To a solution of ethyl 2-(3-ethoxy-1-((6-fluoro-1H-indol-4-yl)amino)-3-oxoprop-1-en-2-yl)-4,5-bis(methoxy-d3)benzoate (480 mg, 1.04 mmol) in THF (14 mL) and H2O (7 mL) was added LiOH·H2O (217.7 mg, 5.19 mmol) and the reaction mixture was stirred at 50° C. overnight. THF was removed in a vacuum. The resulting mixture was acidified with 2 N HCl and the pH was adjusted to 1˜2, a large amount of solid was precipitated. Then the mixture was filtered. The solid was collected, dried to afford 2-(6-fluoro-1H-indol-4-yl)-6,7-bis(methoxy-d3)-1-oxo-1,2-dihydroisoquinoline-4-carboxylic acid (398 mg, 1.02 mmol, 98.7% yield) as a grey solid. LC-MS (ESI): m/z 387.0 [M−H]−.
A mixture of 2-(6-fluoro-1H-indol-4-yl)-6,7-bis(methoxy-d3)-1-oxo-1,2-dihydroisoquinoline-4-carboxylic acid (100 mg, 0.26 mmol) in DMF (10 mL) was added DIEA (0.21 mL, 1.29 mmol),HATU (117.5 mg, 0.31 mmol) and 4-fluoropiperidine (31.9 mg, 0.31 mmol) and the mixture was stirred at RT for 1 h. The mixture was diluted with H2O (100 mL) and the resulting mixture was extracted with EA (50 mL×2). The organic layer was combined, washed with brine (100 mL×3), dried over anhydrous Na2SO4, filtered and concentrated in vacuum to afford the crude 2-(6-fluoro-1H-indol-4-yl)-4-(4-fluoropiperidine-1-carbonyl)-6,7-bis(methoxy-d3)isoquinolin-1(2H)-one (88 mg, 0.19 mmol, 72.2% yield) as a brown solid. LC-MS (ESI): m/z 473.4 [M−H]−.
To a solution of 2-(6-fluoro-1H-indol-4-yl)-4-(4-fluoropiperidine-1-carbonyl)-6,7-bis(methoxy-d3)isoquinolin-1(2H)-one (88 mg, 0.19 mmol) in DMF (10 mL) was added NaH (4.46 mg, 0.19 mmol, 60% in oil) under N2 atmosphere. The resulting mixture was stirred at this temperature for 0.5 hour before CD3I (32.33 mg, 0.23 mmol) was added. The reaction mixture was stirred at RT for 2 hours under N2 atmosphere. The resulting mixture was quenched with H2O (10 mL) and extracted with EA (30 mL*2). The organic layers were combined, washed with brine (30 mL), dried over anhydrous Na2SO4, filtered and concentrated. The residue was purified by silica gel column chromatography (PE/EA=50/1 to 1/1) to afford 2-(6-fluoro-1-(methyl-d3)-1H-indol-4-yl)-4-(4-fluoropiperidine-1-carbonyl)-6,7-bis(methoxy-d3)isoquinolin-1(2H)-one (28.48 mg, 0.06 mmol, 31.2%) as a white solid. LC-MS (ESI): m/z 491.3 [M+H]+. 1H NMR (400 MHz, DMSO-d6) 7.50 (dd, J=9.9, 1.7 Hz, 1H), 7.47 (s, 1H), 7.40 (d, J=3.2 Hz, 1H), 7.11 (dd, J 10.2, 2.0 Hz, 1H), 6.96 (s, 1H), 6.15 (d, J 2.9 Hz, 1H). 4.97-4.81 (m, 1H), 3.89-3.52 (m, 4H), 1.94-1.82 (m, 2H), 1.77-1.68 (m, 2H).
The compounds below were synthesized following the procedures described for Compound 164.
1H NMR (400 MHz, DMSO-d6) δ
To a solution of 2-bromopyridin-3-ol (20 g, 114 mmol) in H2O (250 mL) was added K2CO3 (31.7 g, 229 mmol), I2 (29.6 g, 117 mmol). The reaction mixture was stirred at rt overnight. The mixture was adjusted the pH to 5 with 3N HCl. The resulting solid was collected by filtration, washed with water (20 mL*3), dried to give 2-bromo-6-iodopyridin-3-ol (19.61 g, 65.40 mmol) as a yellow solid. LC-MS (ESI): m/z 299.9 [M+H]+.
To a solution of 2-bromo-6-iodopyridin-3-ol (20 g, 66.6 mmol) in DMF (60 mL) were added K2CO3 (18.4 g, 133 mmol), CH3I (18.9 mL, 233 mmol) and the reaction mixture was stirred at 100° C. for 2 h. Solid precipitation after water (300 mL) was added. The crude was filtered, the cake was washed with water (100 mL*2), dried to afford 2-bromo-6-iodo-3-methoxypyridine (18.4 g, 58.61 mmol) as a yellow solid. LC-MS (ESI): m/z 314.0 [M+H]+.
A mixture of 2-bromo-6-iodo-3-methoxypyridine (6.8 g, 21.6 mmol), methoxysodium (17.3 mL, 86.6 mmol, 4 mol/L in MeOH) and DMSO (30 ml.) was bubbled with argon for 15 scends, then the reaction mixture was stirred at 80° C. for 3 hr. The mixture was diluted with H2O (200 mL), a large amount of solid was precipitated. Then the mixture was filtered. The solid was collected, dried to afford 6-iodo-2,3-dimethoxypyridine (5.8 g, 21.8 mmol) as a brown solid. LC-MS (ESI): m/z 266.0 [M+H]+.
A mixture of 6-iodo-2,3-dimethoxypyridine (5 g, 18.8 mmol), 1,3-diethyl propanedioate (11.51 mL, 75.4 mmol), Cs2CO3 (6.15 g, 18.8 mmol), CuI (3.59 g, 18.8 mmol) and DMF (40 mL) was stirred at 140° C. for 8 h. The mixture was diluted with H2O (100 mL) and the resulting mixture was extracted with EA (100 mL*2). The organic layers were combined, washed with brine (100 mL), dried over anhydrous Na2SO4, filtered and concentrated. The residue was purified by silica gel column chromatography (EA/PE=1/100 to 1/20) to afford 1,3-diethyl 2-(5,6-dimethoxypyridin-2-yl)propanedioate (3 g, 10.09 mmol, 53.5% yield) as a yellow oil. LC-MS (ESI): m/z 298.1 [M+H]+.
To a solution of 1,3-diethyl 2-(5,6-dimethoxypyridin-2-yl)propanedioate (3 g, 10.09 mmol) in DMSO (24 mL) and H2O (4 mL) was added LiCl (2.57 g, 60.54 mmol) and the solution was stirred at 120° C. overnight. The mixture was mixed with H2O (100 mL) and the resulting mixture was extracted with EA (100 mL*2). The organic layers were combined, washed with brine (30 mL), dried over anhydrous Na2SO4, filtered and concentrated in a vacuum. The residue was purified by silica gel column chromatography (PE/EA=20/1 to 6/1) to afford ethyl 2-(5,6-dimethoxypyridin-2-yl)acetate (1 g, 4.44 mmol) as a yellow oil. LC-MS (ESI): m/z 226.1 [M+H]+.
To a solution of ethyl 2-(5,6-dimethoxypyridin-2-yl)acetate (1 g, 4.44 mmol) in CH3CN (20 mL) was added NBS (0.95 g, 5.32 mmol), then the reaction mixture was stirred at rt overnight. The reaction was diluted with EA(50 mL) and Na2SO3 solution (30 mL). The organic layer was separated, washed with further brine (50 mL) and concentrated in a vacuum. The residue was purified using silica gel column chromatography eluting with (EA/PE=0/1-1/10) to afford the title compound ethyl 2-(3-bromo-5,6-dimethoxypyridin-2-yl)acetate (1.20 g, 3.95 mmol) as a yellow oil. LC-MS (ESI): m/z 304.0 [M+H]+.
To a solution of ethyl 2-(3-bromo-5,6-dimethoxypyridin-2-yl)acetate (1.1 g, 3.62 mmol) in MeOH (2 mL) and DMF (2 mL) under CO were added TEA (0.50 mL, 3.61 mmol), Palladium (II) Acetate (3.68 mg, 0.016 mmol), 1,3-bis(diphenylphosphino)propane (0.30 g, 0.723 mmol), and the reaction was stirred at 100° C. for overnight. The residue was purified using silica gel column chromatography eluting with ethyl acetate in petroleum ether (PE/EA: 40/1 ˜1/7) to afford the title compound methyl 2-(2-ethoxy-2-oxoethyl)-5,6-dimethoxypyridine-3-carboxylate (120 mg, 0.42 mmol) as a white solid. LC-MS (ESI): m/z 284.1 [M+H]+.
A mixture of methyl 2-(2-ethoxy-2-oxoethyl)-5,6-dimethoxypyridine-3-carboxylate (120 mg, 0.29 mmol) and [(tert-butoxy)(dimethylamino)methyl]dimethylamine (0.18 mL, 0.86 mmol) was stirred at 100° C. for 3 h. After cooling to rt, to the mixture were added HOAc (5 mL) and 4-fluoro-2-methyl-1H-indol-5-amine (61.4 mg, 0.374 mmol). The reaction was stirred at 50° C. overnight. The mixture was diluted with H2O (50 mL) and the resulting mixture was extracted with EA (100 mL*2). The organic layers were combined, washed with brine (50 mL), dried over anhydrous Na2SO4, filtered and concentrated to afford methyl 2-[(1Z)-3-ethoxy-1-[(4-fluoro-2-methyl-1H-indol-5-yl)amino]-3-oxoprop-1-en-2-yl]-5,6-dimethoxypyridine-3-carboxylate (118 mg, 0.13 mmol) as a brown oil. LC-MS (ESI): m/z 426.2 [M+H]+.
To a solution of ethyl 6-(4-fluoro-2-methyl-1H-indol-5-yl)-2,3-dimethoxy-5-oxo-5,6-dihydro-1,6-naphthyridine-8-carboxylate (118 mg, 0.14 mmol) in THE (8 mL) and H2O (4 mL) was added LiOH·H2O (46.6 mg, 1.11 mmol) and the reaction mixture was stirred at 80° C. for 5 h. The resulting mixture was acidified with 2 N HCl and the pH was adjusted to 1˜2, a large amount of solids was precipitated. Then the mixture was filtered. The solid was collected, dried to afford 6-(4-fluoro-2-methyl-1H-indol-5-yl)-2,3-dimethoxy-5-oxo-5,6-dihydro-1,6-naphthyridine-8-carboxylic acid (77 mg, 0.11 mmol) as a red solid. LC-MS (ESI): m/z 398.1 [M+H]+.
A mixture of 6-(4-fluoro-2-methyl-1H-indol-5-yl)-2,3-dimethoxy-5-oxo-5,6-dihydro-1,6-naphthyridine-8-carboxylic acid (10 mg, 0.03 mmol) in DMF (3 mL) was added DIEA (9.8 mg, 0.08 mmol), HATU (19.1 mg, 0.05 mmol) and 4-fluoropiperidine (3.4 mg, 0.03 mmol) and the mixture was stirred at rt for 1 h. The mixture was diluted with H2O (100 mL) and the resulting mixture was extracted with EA (50 mL*2). The organic layer was combined, washed with brine (100 mL*3), dried over anhydrous Na2SO4, filtered and concentrated in vacuum. The residue was purified using silica gel column chromatography eluting (PE/EA=10:1 to 1:2) to afford the title compound 6-(4-fluoro-2-methyl-1H-indol-5-yl)-8-(4-fluoropiperidine-1-carbonyl)-2,3-dimethoxy-5,6-dihydro-1,6-naphthyridin-5-one (7.81 mg, 0.02 mmol, 64.3% yield) as a white solid. LC-MS (EST): m/z 483.1 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ 11.47 (s, 1H), 7.78 (s, 1H), 7.62 (s, 1H), 7.22 (d, J=8.6 Hz, 1H), 7.08-7.00 (m, 1H), 6.29 (s, 1H), 5.01-4.81 (m, 1H), 3.99 (s, 3H), 3.93 (s, 3H), 3.84-3.59 (m, 2H), 3.53-3.41 (m, 1H), 3.27-3.16 (m, 1H), 2.43 (s, 3H), 1.97-1.60 (m, 4H).
To a solution of 6-(4-fluoro-2-methyl-1H-indol-5-yl)-8-(4-fluoropiperidine-I-carbonyl)-2,3-dimethoxy-5,6-dihydro-1,6-naphthyridin-5-one (45 mg, 0.09 mmol) in DMF (2 mL) was added NaH (11 mg, 0.28 mmol, 60% in oil) under 0° C. The reaction was stirred at this temperature for 30 min before CH3I (0.02 mL, 0.28 mmol) was added. The mixture was stirred at rt for 2 h. The mixture was diluted with H2O (50 mL) and the resulting mixture was extracted with EA (50*3). The organic layer was combined, washed with brine (50 mL), dried over anhydrous Na2SO4, filtered and concentrated in vacuum. The residue was purified by Prep-HPLC (Waters 2767/2545/2489, Waters sunfire C18 10 m OBD 19*250 mm, Mobile Phase A: 0.1% FA in H2O, Mobile Phase B: CH3CN, Flow: 20 mL/min, Column temp: RT) to afford 6-(4-fluoro-1,2-dimethyl-1H-indol-5-yl)-8-(4-fluoropiperidine-1-carbonyl)-2,3-dimethoxy-5,6-dihydro-1,6-naphthyridin-5-one (2.42 mg, 0.00 mmol) as a white solid. LC-MS (ESI): m/z 497.2 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ 7.78 (s, 1H), 7.62 (s, 1H), 7.37 (d, J=8.6 Hz, 1H), 7.23-6.93 (m, 1H), 6.39 (s, 1H), 5.02-4.81 (m, 1H), 3.99 (s, 3H), 3.93 (s, 3H), 3.74 (s, 3H), 3.67-3.23 (m, 4H), 2.45 (s, 3H), 1.87-1.59 (m, 4H).
The compounds below were synthesized following the procedures described for Compound 173.
1H NMR (400 MHz, DMSO-d6) δ
To a solution of 2-chloro-6-methoxypyridine-3-carboxylic acid (5 g, 26.7 mmol) in 1,3-diethyl propanedioate (60 mL, 373 mmol) was added CuBr (380 mg, 2.67 mmol). The reaction was cooled to 0° C. and NaH (2.6 g, 63.9 mmol) was added. Then the mixture was heated to 90° C. for 2 h. The reaction was monitored by TLC. The mixture was diluted with water (50 mL) and extracted with Et2O (30 mL*3). Then the water phase was added 6N HCl to adjust pH=4 and extracted with EA (30 mL*3).The organic layer was separated, washed with further saturated NaCl solution, and concentrated in vacuo to afford 2-(1,3-diethoxy-1,3-dioxopropan-2-yl)-6-methoxynicotinic acid (4 g, 48.2%).
A mixture of 2-(1,3-diethoxy-1,3-dioxopropan-2-yl)-6-methoxynicotinic acid (4 g, 12.8 mmol,) in EtOH (40 mL), H2O (40 mL) and con. HCl (40 mL) was heated to 100° C. for 16 hours. The resulting mixture was concentrated under reduced pressure. The residue was dissolved in EtOH (40 mL) and the whole was cooled to 0° C., then SOCl2 (4 mL) was added dropwise. The resulting mixture was heated to reflux for 3 hours and then concentrated under reduced pressure. The residue was diluted with water (50 mL) and the whole was extracted with EtOAc (20 mL×3). The organic layers were combined, washed with brine, dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel chromatography to afford ethyl 2-(2-ethoxy-2-oxoethyl)-6-hydroxynicotinate (1.8 g, 55%) as a brown solid. LCMS: [M+1]=254
To a solution of ethyl 2-(2-ethoxy-2-oxoethyl)-6-hydroxynicotinate (1.6 g, 6.3 mmol) in CHCl3 (30 mL) was added Ag2CO3 (1.9 g, 6.95 mmol) and MeI (1 g, 7.58 mmol). The mixture was sealed in a tube and stirred at 60° C. for 16 h. The mixture was filtered and concentrated. The residue was purified by flash chromatography to afford ethyl 2-(2-ethoxy-2-oxoethyl)-6-methoxynicotinate (1.4 g, 61.1%) as a white solid. LCMS: [M+1]=268.
To a mixture of ethyl 2-(2-ethoxy-2-oxoethyl)-6-methoxynicotinate (800 mg, 3.0 mmol) in DMF (5 mL) was added 1-tert-butoxy-N,N,N′,N′-tetramethylmethanediamine (2.6 g, 15.0 mmol). The mixture was stirred at 100° C. for 4 hours. The reaction was monitored by TLC. The mixture was dissolved in H2O (20 mL) and extracted with EA (10 mL*2). The organic layers were combined, washed with brine, dried over anhydrous Na2SO4, filtered and concentrated. The residue was dissolved in AcOH (10 mL) and 6-fluoro-1H-indol-4-amine (494 mg, 3.3 mmol) was added. Then the reaction mixture was stirred at RT overnight. The mixture was diluted with H2O (10 mL) and the resulting mixture was extracted with EA (10 mL*2). The organic layers were combined, washed with aq. NaHCO3, dried over anhydrous Na2SO4, filtered and concentrated to afford ethyl 6-(6-fluoro-1H-indol-4-yl)-2-methoxy-5-oxo-5,6-dihydro-1,6-naphthyridine-8-carboxylate (800 mg, 70.1%) as a yellow solid. LCMS: [M+1]-382.
To a solution of ethyl 6-(6-fluoro-1H-indol-4-yl)-2-methoxy-5-oxo-5,6-dihydro-1,6-naphthyridine-8-carboxylate (800 mg, 2.1 mmol) was dissolved in THF/H2O (10 mL/2.5 mL), treated with LiOH (252 mg, 10.5 mmol). The mixture was stirred at 80° C. for 2 hours. TLC showed the reaction was completed. The reaction mixture was concentrated in vacuo. The resulting mixture was acidified with 3N HCl a.q. to pH=3. The precipitated solid was filtered, washed with water. The filter cake was dried under reduced pressure to afford 6-(6-fluoro-1H-indol-4-yl)-2-methoxy-5-oxo-5,6-dihydro-1,6-naphthyridine-8-carboxylic acid (300 mg, 40.5%) as brown solid.
To a solution of 6-(6-fluoro-1H-indol-4-yl)-2-methoxy-5-oxo-5,6-dihydro-1,6-naphthyridine-8-carboxylic acid (300 mg, 0.85 mmol) in DMF (5 mL) was added T3P, 50% in DMF (1.1 g, 1.7 mmol) and DIEA (329 mg, 2.55 mmol). The mixture was stirred at rt for 0.5 h, then piperidine (217 mg, 2.55 mmol) was added. The mixture was stirred at rt for 16 h. The mixture was diluted with H2O (10 mL) and the resulting mixture was extracted with EA (10 mL*2). The organic layers were combined, washed with brine, dried over anhydrous Na2SO4, filtered and concentrated to afford 6-(6-fluoro-1H-indol-4-yl)-2-methoxy-8-(piperidine-1-carbonyl)-1,6-naphthyridin-5(6H)-one (250 mg, 70%) as a white solid. LCMS: [M+1]=421. 1H NMR (400 MHz, CD3OD) δ 8.51 (d, J=8.8 Hz, I H), 7.73 (s, I H), 7.31 (s, 1H), 7.26 (d, J 9.6 Hz, 1H), 7.00 (t, J 8.4 Hz, 2H), 6.25 (s, 1H), 4.06 (s, 3H), 3.77 (s, 2H), 3.43 (s, 2H), 1.66 (d, J 29.6 Hz, 4H), 1.53 (s, 2H).
To a solution of 6-(6-fluoro-I H-indol-4-yl)-2-methoxy-8-(piperidine-1-carbonyl)-1,6-naphthyridin-5(6H)-one (100 mg, 0.238 mmol) in DCE (5 mL) was added cyclopropylboronic acid (81.7 mg, 0.95 mmol), 2,2′-bipyridine (74.3 mg, 0.47 mmol), Na2CO3 (100 mg, 0.95 mmol) and Cu(OAc)2 (172 mg, 0.95 mmol). The mixture was stirred at 70° C. for 16 h. The mixture was diluted with H2O (10 mL) and the resulting mixture was extracted with DCM (10 mL*2). The organic layers were combined, washed with brine, dried over anhydrous Na2SO4, filtered and concentrated. The residue was purified by Prep-HPLC to afford 6-(1-cyclopropyl-6-fluoro-1H-indol-4-yl)-2-methoxy-8-(piperidine-1-carbonyl)-1,6-naphthyridin-5(6H)-one (26.4 mg, 24.1%) as a yellow solid. LCMS: [M+1]=461. 1H NMR (400 MHz, CD3OD) δ 8.50 (d, J=8.8 Hz, 1H), 7.71 (s, 1H), 7.46 (d, J=9.6 Hz, 1H), 7.29 (d, J=3.2 Hz, 1H), 7.05 (d, J=10.0 Hz, 1H), 6.98 (d, J=8.8 Hz, 1H), 6.19 (s, 1H), 4.05 (s, 3H), 3.76 (s, 2H), 3.41 (d, J=6.8 Hz, 3H), 1.65 (d, J=30.8 Hz, 4H), 1.51 (s, 2H), 1.13 (d, J=6.0 Hz, 2H), 1.05-0.93 (m, 2H).
The compounds below were synthesized following the procedures described for Compound 176.
1H NMR (400 MHz, CD3OD) δ
To a solution of ethyl 2-bromo-5,6-dimethoxypyridine-3-carboxylate (20.0 g, 68.94 mmol) in THF (200 mL) was added LiAlH4 (1M in THF, 103.41 mL, 103.41 mmol) dropwise at 0° C. under N2, and the reaction was stirred at 0° C. for 2 hr. Quenched with sat NH4Cl solution, concentrated in vacuo, the residue was purified by silica gel column chromatography eluting with 25% ethyl acetate in petroleum ether to afford the title compound (2-bromo-5,6-dimethoxypyridin-3-yl) methanol (14.37 g, 57.91 mmol, 84%) as a white solid. LC-MS (ESI): m/z 248.1 [M+H+]+
To a solution of (2-bromo-5,6-dimethoxypyridin-3-yl)methanol (14.37 g, 57.92 mmol) in DCM (150 mL) was added SOCl2 (68.29 g, 579.25 mmol) dropwise at 0° C. under N2, and the reaction was stirred at rt for 2 hr. The reaction was diluted with DCM and saturated NaHCO3 solution. The organic layer was separated, washed with brine, and concentrated in vacuo to afford the title compound 2-bromo-3-(chloromethyl)-5,6-dimethoxypyridine (13 g, 48.78 mmol, 84.2%) as a white solid. LC-MS (ESI): m/z 266.1 [M+H]+
To a solution of 2-bromo-3-(chloromethyl)-5,6-dimethoxypyridine (1.0 g, 3.75 mmol) in DMSO (10 mL) was added NaCN (0.20 g, 4.13 mmol) and the reaction was stirred 80° C. temperature for 2 hr. The reaction was diluted with EA and water. The organic layer was separated, washed with brine, and concentrated in vacuo. The residue was purified by silica gel column chromatography eluting with 18% ethyl acetate in petroleum ether to afford the title compound 2-(2-bromo-5,6-dimethoxypyridin-3-yl)acetonitrile (910 mg, 3.54 mmol, 94.3%) as a white solid. LC-MS (ESI): m/z 259.2 [M+H+]+
To a solution of 2-(2-bromo-5,6-dimethoxypyridin-3-yl)acetonitrile (9.9 g, 38.51 mmol) in MeOH (150 mL) were added Pd(dppf)C12 (2.79 g, 3.85 mmol) and TEA (16.0 mL, 115.52 mmol), and the reaction was stirred at 70° C. for 4 hr under 1 atm of CO. Concentrated in vacuo, the residue was purified by silica gel column chromatography eluting with 30% ethyl acetate in petroleum ether to afford the title compound methyl 3-(cyanomethyl)-5,6-dimethoxypyridine-2-carboxylate (6 g, 25.40 mmol, 66.0%) as an off-white solid. LC-MS (ESI): m/z 237.2 [M+H+]+
A solution of methyl 3-(cyanomethyl)-5,6-dimethoxypyridine-2-carboxylate (1.0 g, 4.23 mmol) in DMF-DMA (10 mL) was stirred at 90° C. for 8 hr under N2. The reaction was concentrated in vacuo, and a solution of 4-fluoro-2-methyl-1H-indol-5-amine (0.69 g, 4.23 mmol) in AcOH (10 mL) was added. The reaction was stirred at 50° C. overnight under N2. The reaction was diluted with EA and saturated NaHCO3 solution. The organic layer was separated, washed with brine, and concentrated in vacuo. The residue was purified by silica gel column chromatography eluting with 30%-70% ethyl acetate in petroleum ether to afford methyl 7-(4-fluoro-2-methyl-1H-indol-5-yl)-2,3-dimethoxy-8-oxo-7,8-dihydro-1,7-naphthyridine-5-carbonitrile (840 mg, 2.22 mmol, 52.5%) as a yellow solid. LC-MS (ESI): m/z 379.3 [M+H+]+
To a solution of 7-(4-fluoro-2-methyl-1H-indol-5-yl)-2,3-dimethoxy-8-oxopyrido[3,4-b]pyridine-5-carbonitrile (750 mg, 1.98 mmol) in EtOH (15 mL) was added KOH (2M, 10 mL, 20.00 mmol) and the reaction was stirred at 80° C. overnight. Concentrated in vacuo, the residue was purified by Prep-HPLC to afford the title compound 7-(4-fluoro-2-methyl-1H-indol-5-yl)-2,3-dimethoxy-8-oxopyrido[3,4-b]pyridine-5-carboxylic acid (200 mg, 0.50 mmol, 25.4%) as a brown solid. LC-MS (ESI): m/z 398.3 [M+H+]+
To a solution of 7-(4-fluoro-2-methyl-1H-indol-5-yl)-2,3-dimethoxy-8-oxopyrido[3,4-b]pyridine-5-carboxylic acid (170 mg, 0.43 mmol) in DMF (5 mL) were added 4-fluoropiperidine hydrochloride (5.25 mg, 0.038 mmol), HATU (244.01 mg, 0.64 mmol) and DIEA (331.78 mg, 2.57 mmol), and the reaction was stirred at room temperature for 2 hr. Concentrated in vacuo, the residue was purified by silica gel column chromatography eluting with 60% ethyl acetate in petroleum ether to afford the title compound 5-[(4-fluorohexahydropyridin-1-yl)carbonyl]-7-(4-fluoro-2-methyl-1H-indol-5-yl)-2,3-dimethoxy-7,8-dihydropyrido[3,4-b]pyridin-8-one (100 mg, 0.21 mmol, 48.4%) as a yellow solid. LC-MS (ESI): m/z 483.0 [M+H+]+.
1H NMR (400 MHz, DMSO-d6): δ 11.46 (s, 1H), 7.61 (s, 1H), 7.23-7.21 (m, 2H), 7.06 (t, J=7.2 Hz, 1H), 6.30 (s, 1H), 4.96-4.84 (m, 1H), 4.0 (s, 3H), 3.92 (s, 3H), 3.63-3.55 (m, 4H), 2.43 (s, 3H), 1.93-1.75 (m, 4H). LCMS [mobile phase: from 80% water (0.05% TFA) and 20% acetonitrile to 50% water (0.05% TFA) and 50% acetonitrile in 6.5 min, finally under these conditions for 0.5 min.], Rt=3.547 min; >95% purity; MS Calcd.:482.2; MS Found: 483.0 ([M+H+]+).
To a solution of 5-[(4-fluorohexahydropyridin-1-yl)carbonyl]-7-(4-fluoro-2-methyl-1H-indol-5-yl)-2,3-dimethoxy-7,8-dihydropyrido[3,4-b]pyridin-8-one (60 mg, 0.12 mmol) in DMF (5 mL) was added NaH (14.92 mg, 0.37 mmol) and the reaction was stirred at 0° C. for 30 min under N2, then C3I (0.02 mL, 0.19 mmol) was added in and the reaction was stirred at 0° C. for 1.5 hr. The reaction was diluted with EA and saturated NH4Cl solution. The organic layer was separated, washed with brine, and concentrated in vacuo. The residue was purified by-Prep-HPLC to afford the title compound 7-(4-fluoro-1,2-dimethyl-1H-indol-5-yl)-5-(4-fluoropiperidine-1-carbonyl)-2,3-dimethoxy-1,7-naphthyridin-8(7H)-one(18.3 mg, 0.04 mmol, 29.6%) as a white solid.
1H NMR (400 MHz, DMSO-d6): δ 7.61 (s, 1H), 7.37 (d, J=8.4 Hz, 1H), 7.21 (s, 1H), 7.14 (t, J=7.6 Hz, 1H), 6.39 (s, 1H), 4.96-4.84 (m, 1H), 3.99 (s, 3H), 3.93 (s, 3H), 3.74 (s, 3H), 3.63-3.48 (m, 4H), 2.45 (s, 3H), 1.94-1.75 (m, 4H). LCMS [mobile phase: from 95% water (0.05% TFA) and 5% acetonitrile to 5% water (0.05% TFA) and 95% acetonitrile in 6.5 min, finally under these conditions for 0.5 min.], Rt=3.242 min; >99% purity; MS Calcd.:496.2; MS Found: 497.1 ([M+H+]+).
The compounds below were synthesized following the procedures described for Compound 170.
1H NMR (400 MHz,
Ligand binding to a specific G protein-coupled receptor (GPCR) recruit protease-tagged arrestin proteins to the activated receptor, the protease-tagged arrestin cleaves a protease site fused to the C-terminus of the GPCR, releasing a non-native transcription factor, this transcription factor immediately enters the nucleus, regulating transcription of a beta-lactamase reporter construct, which is measured upon addition of the live-cell substrate.
The H_LPAR5_Tango_CHO_K1 cells (constructed by Genomeditech) was passaged in complete medium (F12K+10% FBS+1% P.S.+4 μg/ml puromycin+4 μg/ml blasticidin+100 μg/ml hygromycin) in an incubator (37° C., 5% CO2) were used in the Tango screening assay.
Prior to the Tango assay, CHO K1 cells were kept for 6 h in FBS-depleted media (DMEM, 0.10% BSA, 10% penicillin-streptomycin, 4 μg/ml puromycin+4 μg/ml blasticidin+100 μg/ml hygromycin).
Approximately 1*104 H_LPAR5-Tango Reporter CHO K1 cells were seeded into a 96-well flat, white-bottom assay plate with 100 ul complete medium overnight. Next day the medium was changed to starving medium (F12K, 0.1% BSA, 1% penicillin-streptomycin, 0.75 g/mL Puromycin, 4 μg/mL Blasticidin, 400 μg/ml G418). After starvation for 6 h, the compounds at desired concentration (3× dilution) were added into each well and incubated with cells at 37° C. with 5% CO2. One hour later, 5 μM LPA was added into each well and incubated with cells at 37° C. for 12 h. Then the medium was removed and the cell plate was put into −80° C. refrigerator for 1 h for cell lysis. After that, the cell plate was put at room temperature for 45 min for equilibration, then 45 ul ONE-Glo Luciferase Assay reagent (ONE-Glo reagent: F12K (no FBS)=1:2, mix well) was added, and kept in dark for 10 min at room temperature, then read plate using microplate luminescence reader (405 nm).
Intracellular calcium release stimulated by a GPCR agonist initiates an important signaling pathway that triggers a variety of cellular functions including downstream calcium-dependent transcription factors, such as nuclear factor of activated T-cells (NFAT) that regulates luciferase gene expression. Combined with ONE-Glo™ Luciferase Assay System, the LPAR5 antagonists screening assay were evaluated in vitro.
The HEK293 cell line was used to monitor the inhibitory effect of antagonists on NFAT transcriptional activity. All the compounds are investigated based on the following assay methods.
HEK293T cells were passaged in complete medium (DMEM, 10% FBS, 1% penicillin-streptomycin, 0.75 pg/mL Puromycin, 4 g/mL Blasticidin, 400 μg/ml G418) in incubator (37° C., 5% CO2). Prior to the NFAT assay, HEK293T cells were kept for 6 h in FBS-depleted media (DMEM, 0.1% BSA, 1% penicillin-streptomycin, 0.75 μg/mL Puro, 4 g/mL Blast, 400 μg/ml G418).
Approximately 1.5*104 H-GNA15-NFAT Reporter HEK-293 cells were seeded into 96-well flat, white-bottom assay plate with 100 μl complete medium over night. Next day the medium was changed to starving medium (DMEM, 0.10% BSA, 10% penicillin-streptomycin, 0.75 μg/mL Puromycin, 4 g/mL Blasticidin, 400 μg/ml G418). After starvation for 6 h, the compounds disclosed herein at desired concentration (10 uM) were added into each well and incubated with cells at 37° C. with 5% CO2. One hour later, 5 μM LPA was added into each well and incubated with cells at 37° C. for 12 h. Then the medium was removed and the cell plate was put into −80° C. refrigerator for 1h for cell lysis. After that, the cell plate was put in room temperature for 45 min for equilibration, then 45 ul ONE-Glo Luciferase Assay reagent (ONE-Glo reagent: F12K (no FBS)=1:2, mix well) was added, and kept in dark for 10 min at room temperature, then read plate using microplate luminescence reader (405 nm)
Thaw liver microsome in 37° C. water bath and make liver microsome working solution for each compound based on following calculations:
17 mg/mL NADPH and 19 mg/mL UDPGA working solutions were prepared with phosphate buffer and then combine equal volumes of each to get the final co-factor working solution. Liver microsome working solution was pre-incubated at 37° C. for 5 min and was added 1.5 μL control/the test compound working solution to 238.5 μL liver microsome working solution, and the resultant mixture was gently mixed. The mixture was pre-incubated at 37° C. for 5 min and the reaction was initiated by adding 60 μL. of the co-factor working solution. The mixture was pipetted up and down. At each time point, 30 μL reaction mixture was removed, and 300 μL quenching solution was added. The mixture was vigorously vortexed for 1 min and centrifuged at 4,000 rpm at 4° C. for 15 min. 100 μL of the supernatant was drew up and mixed with 100 μL distilled water for LC-MS/MS analysis.
T1/2 was calculated by following equations:
The results of the biological test are shown below.
Although the foregoing invention has been described in some detail by way of illustration and example for purposes of clarity of understanding, it is apparent to those skilled in the art that certain minor changes and modifications will be practiced. Therefore, the description and examples should not be construed as limiting the scope of the invention.
| Number | Date | Country | Kind |
|---|---|---|---|
| PCT/CN2022/079039 | Mar 2022 | WO | international |
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/CN2023/079498 | 3/3/2023 | WO |
