Pyridazinones and methods of use thereof

Abstract

To provide therapeutic methods for treating kidney diseases.SOLUTION: There are provided therapeutic methods for treating kidney diseases, using compounds of Formula (A) in combination with a second therapeutic agent. (In the formula: each R is independently selected from a group comprising H, alkyl, alkenyl, alkynyl, aryl, heterocyclyl, heteroaryl, halogen, -OH, CN, cycloalkyl, -O-alkyl, -O-cycloalkyl, -O-aryl, -aryl-O-aryl, -CF3, -C(H)F2, alkylene-CF3, alkylene-C(H)F2, -SO2-alkyl, -O-alkylene-O-alkyl, -heterocyclyl-L-R4, and heteroaryl-L-R4; and only one R is -heterocyclyl-L-R4 or -heteroaryl-L-R4). The second therapeutic agent is selected from an immunomodulator, a calcineurin inhibitor, a renin angiotensin aldosterone system inhibitor, an antiproliferative agent, an alkylating agent, and the like.SELECTED DRAWING: None

Description

[Technical Field]

[0001] Related Applications This application claims the benefit of priority to U.S. Provisional Patent Application No. 62 / 821,178, filed March 20, 2019. [Background technology]

[0002] Proteinuria is a condition in which excessive amounts of blood protein leak into the urine. Proteinuria can progress from a loss of 30 mg of protein in the urine per 24 hours (called microalbuminuria) to a loss of more than 300 mg / day (called macroalbuminuria), and then reach levels of more than 3.5 grams per 24 hours, or 25 times the normal amount. Proteinuria occurs when the kidney's glomeruli malfunction, causing fluid accumulation (edema) in the body. Long-term protein leakage has been shown to lead to kidney failure. Nephrotic syndrome (NS) accounts for approximately 12% of end-stage renal disease cases and accounts for an annual cost of more than $3 billion in the United States. Approximately 5 per 100,000 children are diagnosed with NS each year, and 15 per 100,000 children currently suffer from NS. Even in patients who respond positively to treatment, recurrence is extremely common. While 90% of children with NS respond to treatment, an estimated 75% experience recurrence. There is a need for more effective methods of treating or reducing the risk of developing kidney disease (eg, proteinuria).

[0003] Mammalian TRP channel proteins form six-transmembrane, cation-permeable channels that can be classified into six subfamilies (TRPC, TRPV, TRPM, TRPA, TRPP, and TRPML) based on amino acid sequence homology. Recent research on TRP channels has demonstrated that they are involved in many fundamental cellular functions and likely play an important role in the pathophysiology of many diseases. Many TRPs are expressed along different parts of the nephron in the kidney, and increasing evidence suggests that these channels are involved in inherited as well as acquired kidney disorders. TRPC6, TRPM6, and TRPP2 are thought to be involved in hereditary focal segmental glomerulosclerosis (FSGS), hypomagnesemia with secondary hypocalcemia (HSH), and polycystic kidney disease (PKD), respectively.

[0004] TRPC5 has also been reported to contribute to the mechanisms underlying the regulation of innate fear responses (J Neurosci. 2014 Mar 5;34(10):3653-3667). Summary of the Invention

[0005] One aspect of the present invention is a method for treating kidney disease, comprising co-administering to a subject in need thereof a TRPC5 inhibitor compound and a second therapeutic agent. In some embodiments, the method comprises co-administering to a subject in need thereof: a. A TRPC5 inhibitor compound of structural formula (A), or a tautomer or pharmaceutically acceptable salt thereof

[0006] [ka] (In the formula, Each R is independently H, alkyl, alkenyl, alkynyl, aryl, heterocyclyl, heteroaryl, halogen, -OH, CN, cycloalkyl, -O-alkyl, -O-cycloalkyl, -O-aryl, -aryl-O-aryl, -CF3, -C(H)F2, alkylene-CF3, alkylene-C(H)F2, -SO2-alkyl, -O-alkylene-O-alkyl, -heterocyclyl-LR 4 , and heteroaryl-LR 4 selected from the group consisting of R 4 is absent or is an alkyl, cycloalkyl, polycyclyl, aryl, heterocyclyl, heteroaryl, -C(O)N(R 5 CF3, R 5 are independently H or alkyl; R 6 is an alkyl, cycloalkyl, aryl, heterocyclyl, heteroaryl, alkylene-aryl, -C(O)N(R 5 CF3, L is absent or is methylene, -C(O)-, -SO2-, -CH2N(Me)-, -N(R 5 )(R 6 )-, -C(R 5 )(R 6 )-, and -OR 6 selected from the group consisting of If only one R is -heterocyclyl-LR 4 or -heteroaryl-LR 4 ) and b. A second therapeutic agent selected from the following: immunomodulators, calcineurin inhibitors, renin-angiotensin-aldosterone system inhibitors, antiproliferative agents, alkylating agents, corticosteroids, angiotensin-converting enzyme inhibitors, adrenocorticotropic hormone stimulators, angiotensin receptor blockers, sodium-glucose transport protein 2 inhibitors, dual sodium-glucose transport protein 1 / 2 inhibitors, nuclear factor-1 (erythroid-derived 2)-like 2 agonists, chemokine receptor 2 inhibitors, chemokine receptor 5 inhibitors, endothelin 1 receptor antagonists, beta-blockers, mineralocorticoid receptor antagonists, loop or thiazide diuretics, calcium channel blockers, statins, short-, intermediate-, or long-acting insulins, dipeptidyl peptidase 4 inhibitors, glucagon-like peptide 1 receptor agonists, sulfonylureas, apoptosis signal-regulating kinase-1, chymase inhibitors, and selective glycation inhibitors. substances, renin inhibitors, interleukin-33 inhibitors, farnesoid X receptor agonists, soluble guanylate cyclase stimulators, thromboxane receptor antagonists, xanthine oxidase inhibitors, erythropoietin receptor agonists, cannabinoid receptor type 1 inverse agonists, NADPH oxidase inhibitors, anti-vascular endothelial growth factor B, anti-fibrotic agents, neprilysin inhibitors, dual CD80 / CD86 inhibitors, CD40 antagonists, cell-collection inhibitors Terol and lipid blockers, PDGFR antagonists, Slit guidance ligand 2, APOL1 inhibitors, Nrl2 activators / NF-κB inhibitors, somatostatin receptor agonists, PPAR gamma agonists, AMP-activated protein kinase stimulators, tyrosine kinase inhibitors, glucosylceramide synthase inhibitors, arginine vasopressin receptor 2 antagonists, xanthine oxidase inhibitors, and vasopressin receptor 2 antagonists.

[0007] In some embodiments, the TRPC5 inhibitor and the second therapeutic agent are administered as separate dosage forms.

[0008] In an alternative embodiment, the TRPC5 inhibitor and the second therapeutic agent are administered together as a fixed dose combination (ie, a single formulation).

[0009] In some embodiments, the second therapeutic agent is an immunomodulator, a calcineurin inhibitor, a renin-angiotensin-aldosterone system inhibitor, an antiproliferative agent, a corticosteroid, an angiotensin-converting enzyme inhibitor, an angiotensin receptor blocker, a sodium-glucose transport protein 2 inhibitor, a nuclear factor-1 (erythroid-derived 2)-like 2 agonist, a chemokine receptor 2 inhibitor, a chemokine receptor 5 inhibitor, or an endothelin 1 receptor antagonist.

[0010] In some embodiments, the TRPC5 inhibitor compound is represented by structural formula (AI), (A-II), or (A-III), or a tautomer or pharmaceutically acceptable salt thereof:

[0011] [ka] (In the formula, R 1 and R 3 are independently H, alkyl, alkenyl, alkynyl, aryl, heterocyclyl, heteroaryl, halogen, —OH, CN, cycloalkyl, —O-alkyl, —O-cycloalkyl, —O-aryl, -aryl-O-aryl, —CF, —C(H)F, alkylene-CF, alkylene-C(H)F, —SO-alkyl, and —O-alkylene-O-alkyl, -heterocyclyl-LR 4 , and heteroaryl-LR 4 selected from the group consisting of R 2 -heterocyclyl-LR 4 and R 4 is absent or is selected from alkyl, cycloalkyl, aryl, alkylene-aryl, alkylene-heteroaryl, heteroaryl, heterocyclyl, -C(O)N(R 5CF3, R 5 are independently H or alkyl; R 6 is an alkyl, cycloalkyl, aryl, heterocyclyl, heteroaryl, alkylene-aryl, -C(O)N(R 5 CF3, L is absent or is methylene, -C(O)-, -SO2-, -CH2N(Me)-, -N(R 5 )(R 6 )-, -C(R 5 )(R 6 )-, and -OR 6 selected from the group consisting of R 1 , R 2 , and R 3 and only one of the groups is -heterocyclyl-LR 4 or -heteroaryl-LR 4 (It is).

[0012] In some embodiments, the TRPC5 inhibitor compound has structural formula (I):

[0013] [ka] or a pharmaceutically acceptable salt thereof, wherein "---" is a single or double bond, X 1 is CH or N, If "---" is a double bond, X 2 is CH or N, If "---" is a single bond, X 2 is N(CH3), X 1 If is CH, then X 2 is N or N(CH3), Y is -O-, -N(CH3)-, -N(CH2CH2OH)-, cyclopropane-1,1-diyl, or -CH(CH3)-; Q is 2-trifluoromethyl-4-fluorophenyl, 2-difluoromethyl-4-fluorophenyl, 2-trifluoromethylphenyl, 2-methyl-4-fluorophenyl, 2-chloro-4-fluorophenyl, 2-chlorophenyl, 1-(benzyl)-4-methylpiperidin-3-yl, 4-trifluoromethylpyridin-3-yl, 2-trifluoromethyl-6-fluorophenyl, 2-trifluoromethyl-3-cyanophenyl, 2-ethyl-3-fluorophenyl, 2-chloro-3-cyanophenyl, 2-trifluoromethyl-5-fluorophenyl, or 2-difluoromethylphenyl; If "---" is a double bond, R 13 is hydrogen, -CH2OH, -CH(OH)-CH2OH, -NH2, -CH(OH)CH3, -OCH3, or -NH-(CH2)2OH, and R 14 does not exist, or If "---" is a single bond, R 13 and R 14 together form =O, R 5 and R 6 each is independently hydrogen or —CH3).

[0014] In some embodiments, the TRPC5 inhibitor compound has structural formula (II):

[0015] [ka] or a pharmaceutically acceptable salt thereof, wherein R 11 is chloro, -CF3, -CHF2, or -CH3, R 12 is hydrogen or fluoro, R 3 is hydrogen, -NH2, -CH2OH, or CH(OH)-CH2OH).

[0016] In some embodiments, the immunomodulator is rituximab. In some embodiments, the angiotensin-converting enzyme inhibitor is captopril, zofenopril, enalapril, ramipril, quinapril, perindopril, lisinopril, benazepril, imidapril, trandolapril, or cilazapril.

[0017] In some embodiments, the angiotensin receptor blocker is losartan, candesartan, valsartan, irbesartan, telmisartan, eprosartan, olmesartan, azilsartan, or fimasartan.

[0018] In some embodiments, the renin-angiotensin-aldosterone system inhibitor is aliskiren.

[0019] In some embodiments, the endothelin 1 receptor antagonist is ambrisentan, atrasentan, bosentan, or sparsentan. In some additional embodiments, the endothelin 1 receptor antagonist is macitentan.

[0020] In some embodiments, the antiproliferative agent is mycophenolate mofetil, hi some additional embodiments, the antiproliferative agent is mycophenolate sodium or azathioprine.

[0021] In some embodiments, the SGLT2 inhibitor is canagliflozin, dapagliflozin, empagliflozin, a combination of empagliflozin and linagliptin, a combination of empagliflozin and metformin, or a combination of dapagliflozin and metformin. In some additional embodiments, the SGLT2 inhibitor also inhibits SGLT1. In some aspects of these embodiments, the SGLT1 / 2 inhibitor is sotagliflozin.

[0022] In some embodiments, the calcineurin inhibitor is cyclosporin A or tacrolimus. In some additional embodiments, the calcineurin inhibitor is voclosporin.

[0023] In some embodiments, the nuclear factor-1 (erythroid-derived 2)-like 2 agonist is bardoxolone or CXA-10.

[0024] In some embodiments, the chemokine receptor 2 inhibitor is PF-04136309 or ccx140. In some additional embodiments, the chemokine receptor 2 inhibitor is propagemanium (DMX-200).

[0025] In some embodiments, the beta blocker is metoprolol succinate, metoprolol tartrate, propranolol, or carvedilol.

[0026] In some embodiments, the mineralocorticoid receptor antagonist is spironolactone, eplerenone, finerenone, esaxerenone, or aparalenone.

[0027] In some embodiments, the loop or thiazide diuretic is furosemide, bumetanide, torasemide, or bendroflumethiazide.

[0028] In some embodiments, the calcium channel blocker is verapamil, diltiazem, amlodipine, or nifedipine.

[0029] In some embodiments, the statin is atorvastatin, pravastatin, fluvastatin, lovastatin, rosuvastatin, simvastatin, or pitavastatin.

[0030] In some embodiments, the short-intermediate or long-acting insulin is NPH insulin (Humulin®, Novolin®, or a biosimilar), insulin lispro (Humalog®), insulin glulisine, insulin glargine (Basaglar®, Lantus®), insulin detemir (Levemir®), or insulin degludec (Tresiba®).

[0031] In some embodiments, the dipeptidyl peptidase 4 inhibitor is sitagliptin, saxagliptin, linagliptin, or vildagliptin.

[0032] In some embodiments, the glucagon-like peptide 1 receptor agonist is exenatide, liraglutide, dulaglutide, lixisenatide, albiglutide, or semaglutide.

[0033] In some embodiments, the sulfonylurea is glimepiride, glipizide, glyburide, glibenclamide, chlorpropamide, tolazamide, or tolbutamide.

[0034] In some embodiments, the apoptosis signal-regulating kinase-1 is selonsertib.

[0035] In some embodiments, the chymase inhibitor is furasymstat (BAY1142524).

[0036] In some embodiments, the selective glycation inhibitor is GLY-230.

[0037] In some embodiments, the renin inhibitor is SCO-272.

[0038] In some embodiments, the interleukin-33 inhibitor is MEDI-3506.

[0039] In some embodiments, the farnesoid X receptor agonist is nidufexor (LMB763).

[0040] In some embodiments, the soluble guanylate cyclase stimulator is praliciguat, olinciguat, IW-6463, vericiguat, or riociguat.

[0041] In some embodiments, the thromboxane receptor antagonist is SER150.

[0042] In some embodiments, the xanthine oxidase inhibitor is TMX-049.

[0043] In some embodiments, the erythropoietin receptor agonist is sibinetide (ARA-290).

[0044] In some embodiments, the cannabinoid receptor type 1 inverse agonist is nimasimab, GFB-024, or CRB-4001.

[0045] In some embodiments, the NADPH oxidase inhibitor is APX-115.

[0046] In some embodiments, the anti-vascular endothelial growth factor B is CSL-346.

[0047] In some embodiments, the anti-fibrotic agent is FT011.

[0048] In some embodiments, the neprilysin inhibitor is TD-1439, TD-0714, or sacubitril.

[0049] In some embodiments, the dual CD80 / CD86 inhibitor is abatacept.

[0050] In some embodiments, the CD40 antagonist is bleselumab (ASKP1240).

[0051] In some embodiments, the cellular cholesterol and lipid blocker is VAR-200.

[0052] In some embodiments, the PDGFR antagonist is ANG_3070.

[0053] In some embodiments, the Slit guidance ligand 2 is PF-06730512.

[0054] In some embodiments, the APOL1 inhibitor is VX-147.

[0055] In some embodiments, the Nrl2 activator / NF-κB inhibitor is bardoxolone.

[0056] In some embodiments, the somatostatin receptor agonist is lanreotide.

[0057] In some embodiments, the PPAR gamma agonist is pioglitazone.

[0058] In some embodiments, the AMP-activated protein kinase stimulator is metformin.

[0059] In some embodiments, the tyrosine kinase inhibitor is tesevatinib.

[0060] In some embodiments, the glucosylceramide synthase inhibitor is benglustat malate.

[0061] In some embodiments, the arginine vasopressin receptor 2 antagonist is lixivaptan.

[0062] In some embodiments, the xanthine oxidase inhibitor is oxypurinol.

[0063] In some embodiments, the vasopressin receptor 2 antagonist is tolvaptan.

[0064] In some embodiments, the second therapeutic agent is tacrolimus, cyclosporin A, rituximab, mycophenolate mofetil, a corticosteroid, sparsentan, enalapril, or losartan.

[0065] In some embodiments, the disease or condition is focal segmental glomerulosclerosis (FSGS), primary focal segmental glomerulosclerosis, hereditary focal segmental glomerulosclerosis, secondary focal segmental glomerulosclerosis, diabetic nephropathy, Alport syndrome, hypertensive kidney disease, nephrotic syndrome, steroid-resistant nephropathy, minimal change nephrotic syndrome, membranous nephropathy, idiopathic membranous nephropathy, membranoproliferative glomerulonephritis (MPGN), immune complex-mediated MPGN, complement-mediated MPGN, lupus nephritis, post-infectious glomerulonephritis, thin basement membrane disease, mesangial proliferative glomerulonephritis, amyloidosis (primary), c1q nephritis, rapidly progressive GN, anti-GBM disease, C3 glomerulonephritis, hypertensive nephrosclerosis, or IgA nephropathy. In some embodiments, the disease or condition is focal segmental glomerulosclerosis.

[0066] The methods are effective in a variety of subjects, including mammals, e.g., humans, and other animals, e.g., laboratory animals, e.g., mice, rats, rabbits, or monkeys, or pets and livestock, e.g., cats, dogs, goats, sheep, pigs, cows, or horses. In some embodiments, the subject is a human.

[0067] The present invention provides several advantages. The prophylactic and therapeutic methods described herein are effective in treating kidney disease (e.g., proteinuria) with minimal, if any, side effects. Furthermore, the methods described herein are effective in identifying compounds that treat or reduce the risk of developing kidney disease, anxiety, depression, or cancer.

[0068] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although methods and materials similar or equivalent to those described herein can be used in practicing or testing the present invention, suitable methods and materials are described below. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, the present specification, including definitions, will control. Furthermore, the materials, methods, and examples are illustrative only and are not intended to be limiting.

[0069] Other features, objects, and advantages of the invention will be apparent from the detailed description and claims. [Brief explanation of the drawings]

[0070] [Figure 1] 1 shows albumin excretion in PAN-injured rats treated with Compound 100 or mizoribine. [Figure 2] This shows angiogenesis when human kidney organoids were transplanted under the renal capsule of a rat. [Figure 3] Oral administration of Compound 100 results in drug exposure of transplanted organoids. [Figure 4] 1 shows a plot of the effect of Compound AO on albumin excretion in DOCA-salt hypertensive rats. [Figure 5A] Confocal microscopy images of mouse podocytes pretreated with DMSO are shown. [Figure 5B] 1 shows confocal microscopy images of mouse podocytes treated with protamine sulfate (PS). [Figure 5C] Quantification of podocytes with disrupted actin cytoplasm after invasion with PS is shown. [Figure 5D] Confocal microscopy images of mouse podocytes pretreated with compound AO and invaded with PS are shown. [Figure 5E] Confocal microscopy images of mouse podocytes pretreated with compound AO and invaded with PS are shown. [Figure 5F] Confocal microscopy images of mouse podocytes pretreated with compound AO and invaded with PS are shown. [Figure 6A] Confocal microscopy images of human iPSC-derived kidney organoids pretreated with DMSO are shown. [Figure 6B] Confocal microscopy images of human iPSC kidney organoids invaded with protamine sulfate (PS) are shown. [Figure 6C] Quantification of mean phalloidin intensity per organoid after insult with PS is shown. [Figure 6D] Confocal microscopy images of human iPSC kidney organoids pretreated with compound AO and invaded with PS are shown. [Figure 6E] Confocal microscopy images of human iPSC kidney organoids pretreated with compound AO and invaded with PS are shown. [Figure 6F] Confocal microscopy images of human iPSC kidney organoids pretreated with compound AO and invaded with PS are shown. DETAILED DESCRIPTION OF THE INVENTION

[0071] definition The term "acyl" is art-recognized and refers to a group represented by the general formula: hydrocarbylC(O)-, preferably alkylC(O)-.

[0072] The term "acylamino" is art-recognized and refers to an amino group substituted with an acyl group and may be represented, for example, by the formula: hydrocarbylC(O)NH-.

[0073] The term "acyloxy" is art-recognized and refers to a group represented by the general formula: hydrocarbylC(O)O-, preferably alkylC(O)O-.

[0074] The term "alkoxy" refers to an alkyl group, preferably a lower alkyl group, having an oxygen attached thereto. Representative alkoxy groups include methoxy, trifluoromethoxy, ethoxy, propoxy, tert-butoxy, and the like.

[0075] The term "alkoxyalkyl" refers to an alkyl group substituted with an alkoxy group and can be represented by the general formula: alkyl-O-alkyl.

[0076] As used herein, the term "alkenyl" refers to an aliphatic group containing at least one double bond and is intended to include both "unsubstituted alkenyl" and "substituted alkenyl," the latter of which refers to an alkenyl moiety having substituents replacing a hydrogen on one or more carbon atoms of the alkenyl group. Such substituents may be on one or more carbons included or not included in one or more double bonds. Furthermore, such substituents include all of those contemplated for alkyl groups, as described below, except where stability would be inhibited. For example, substitution of alkenyl groups with one or more alkyl, carbocyclyl, aryl, heterocyclyl, or heteroaryl groups is contemplated.

[0077] An "alkyl" group or "alkane" is a completely saturated, straight-chain or branched, non-aromatic hydrocarbon. Typically, a straight-chain or branched alkyl group has from 1 to about 20, preferably from 1 to about 10, carbon atoms, unless otherwise defined. Examples of straight-chain and branched alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, pentyl, hexyl, pentyl, and octyl. C1-C6 straight-chain or branched alkyl groups are also referred to as "lower alkyl" groups.

[0078] Furthermore, as used throughout the specification, examples, and claims, the term "alkyl" (or "lower alkyl") is intended to include both "unsubstituted alkyl" and "substituted alkyl," the latter of which refers to an alkyl moiety having substituents replacing a hydrogen on one or more carbons of the hydrocarbon backbone. Such substituents can include, for example, halogen (e.g., fluoro), hydroxyl, carbonyl (e.g., carboxyl, alkoxycarbonyl, formyl, or acyl), thiocarbonyl (e.g., thioester, thioacetate, or thioformate), alkoxyl, phosphoryl, phosphate, phosphonate, phosphinate, amino, amido, amidine, imine, cyano, nitro, azido, sulfhydryl, alkylthio, sulfate, sulfonate, sulfamoyl, sulfonamido, sulfonyl, heterocyclyl, aralkyl, or aromatic or heteroaromatic moieties, unless otherwise specified. In a preferred embodiment, the substituent on a substituted alkyl is C 1~6 Alkyl, C 3~6 The substituents on the substituted alkyl are selected from cycloalkyl, halogen, carbonyl, cyano, or hydroxyl. In a more preferred embodiment, the substituents on the substituted alkyl are selected from fluoro, carbonyl, cyano, or hydroxyl. Those skilled in the art will understand that, where appropriate, the moieties substituted on the hydrocarbon chain can themselves be substituted. For example, substituents on the substituted alkyl can include amino, azido, imino, amido, phosphoryl (including phosphonates and phosphinates), sulfonyl (including sulfates, sulfonamides, sulfamoyl, and sulfonates), and silyl groups, as well as substituted and unsubstituted forms of ethers, alkylthio, carbonyl (including ketones, aldehydes, carboxylates, and esters), -CF3, -CN, and the like. Exemplary substituted alkyls are described below. Cycloalkyls can be further substituted with alkyls, alkenyls, alkoxys, alkylthios, aminoalkyls, carbonyl-substituted alkyls, -CF3, -CN, and the like.

[0079] Unless otherwise specified, "alkylene," by itself or as part of another substituent, refers to a saturated, straight-chain or branched divalent radical having the stated number of carbon atoms and derived from the corresponding alkane by removing two hydrogen atoms. Examples of straight-chain and branched alkylene groups include -CH-(methylene), -CH-CH-(ethylene), -CH-CH-CH-(propylene), -C(CH)-, -CH-CH(CH)-, -CH-CH-CH-CH-, -CH-CH-CH-CH-CH-(pentylene), -CH-CH(CH)-CH-, and -CH-C(CH)-CH-.

[0080] "C x~y The term "C" when used with a chemical moiety such as acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy is meant to include groups containing x to y carbons in the chain. For example, "C x~y The term "alkyl" refers to substituted or unsubstituted saturated hydrocarbon groups, including straight-chain alkyl and branched alkyl groups, containing x to y carbons in the chain, including haloalkyl groups. Preferred haloalkyl groups include trifluoromethyl, difluoromethyl, 2,2,2-trifluoroethyl, and pentafluoroethyl. CO alkyl refers to hydrogen when the group is in a terminal position and to a bond when internal. "C 2~y alkenyl" and "C 2~y The term "alkynyl" refers to substituted or unsubstituted unsaturated aliphatic groups analogous in length and possible substitution to the alkyls described above, but which contain at least one double or triple bond respectively.

[0081] As used herein, the term "alkylamino" refers to an amino group substituted with at least one alkyl group.

[0082] As used herein, the term "alkylthio" refers to a thiol group substituted with an alkyl group and can be represented by the general formula: alkylS-.

[0083] As used herein, the term "alkynyl" refers to an aliphatic group containing at least one triple bond and is intended to include both "unsubstituted alkynyl" and "substituted alkynyl," the latter of which refers to an alkynyl moiety having substituents replacing a hydrogen on one or more carbon atoms of the alkynyl group. Such substituents may be on one or more carbons included or not included in one or more triple bonds. Furthermore, such substituents include all of those contemplated for alkyl groups as described above, except where stability would be inhibited. For example, substitution of alkynyl groups with one or more alkyl, carbocyclyl, aryl, heterocyclyl, or heteroaryl groups is contemplated.

[0084] As used herein, the term "amide" refers to the group:

[0085] [ka] (wherein each R A independently represent hydrogen or a hydrocarbyl group, or two R A together with the N atom to which they are attached complete a heterocycle with 4 to 8 atoms in the ring structure).

[0086] The terms "amine" and "amino" are art-recognized and refer to both unsubstituted and substituted amines, as well as salts thereof, e.g.,

[0087] [ka] where each R A independently represent hydrogen or a hydrocarbyl group, or two R A together with the N atom to which they are attached complete a heterocycle with 4 to 8 atoms in the ring structure).

[0088] As used herein, the term "aminoalkyl" refers to an alkyl group substituted with an amino group.

[0089] As used herein, the term "aralkyl" refers to an alkyl group substituted with an aryl group.

[0090] As used herein, the term "aryl" includes substituted or unsubstituted monocyclic aromatic groups, in which each atom of the ring is carbon. Preferably, the ring is a 6- or 10-membered ring, more preferably a 6-membered ring. The term "aryl" also includes polycyclic ring systems having two or more rings in which two or more carbons are common to two adjacent rings, where at least one of the rings is aromatic, and the other rings may be, for example, cycloalkyl, cycloalkenyl, aryl, heteroaryl, and / or heterocyclyl. Aryl groups include benzene, naphthalene, phenanthrene, phenol, aniline, and the like.

[0091] The term "carbamate" is art-recognized and refers to a group:

[0092] [ka] (wherein each R A independently represent hydrogen or a hydrocarbyl group (e.g., an alkyl group), or both R A together with the intervening atom(s) complete a heterocycle having 4 to 8 atoms in the ring structure).

[0093] As used herein, the terms "carbocycle" and "carbocyclic" refer to saturated or unsaturated rings in which each atom of the ring is carbon. The term carbocycle includes both aromatic and non-aromatic carbocycles. Non-aromatic carbocycles include both cycloalkane rings in which all carbon atoms are saturated and cycloalkene rings with at least one double bond. "Carbocycle" includes 5- to 7-membered monocyclic and 8- to 12-membered bicyclic rings. Each ring of a bicyclic carbocycle may be selected from saturated, unsaturated, and aromatic rings. Carbocycles include bicyclic molecules in which two rings share one, two, three, or more atoms. The term "fused carbocycle" refers to a bicyclic carbocycle in which each ring shares two adjacent atoms with the other ring. Each ring of a fused carbocycle may be selected from saturated, unsaturated, and aromatic rings. In an exemplary embodiment, an aromatic ring (e.g., phenyl) may be fused with a saturated or unsaturated ring (e.g., cyclohexane, cyclopentane, or cyclohexane). Any combination of saturated, unsaturated, and aromatic bicyclic rings is included in the definition of carbocycle, as long as valence allows. Exemplary "carbocycles" include cyclopentane, cyclohexane, bicyclo[2.2.1]heptane, 1,5-cyclooctadiene, 1,2,3,4-tetrahydronaphthalene, bicyclo[4.2.0]oct-3-ene, naphthalene, and adamantane. Exemplary fused carbocycles include decalin, naphthalene, 1,2,3,4-tetrahydronaphthalene, bicyclo[4.2.0]octane, 4,5,6,7-tetrahydro-1H-indene, and bicyclo[4.1.0]hept-3-ene. A "carbocycle" can be substituted at any one or more positions capable of bearing a hydrogen atom.

[0094] A "cycloalkyl" group is a fully saturated cyclic hydrocarbon. "Cycloalkyl" includes monocyclic and bicyclic rings. Typically, a monocyclic cycloalkyl group has 3 to about 10 carbon atoms, more typically 3 to 8 carbon atoms, unless otherwise defined. The second ring of a bicyclic cycloalkyl can be selected from saturated, unsaturated, and aromatic rings. Cycloalkyl includes bicyclic molecules in which two rings share one, two, three, or more atoms. The term "fused cycloalkyl" refers to a bicyclic cycloalkyl in which each ring shares two adjacent atoms with the other ring. The second ring of a fused bicyclic cycloalkyl can be selected from saturated, unsaturated, and aromatic rings. A "cycloalkenyl" group is a cyclic hydrocarbon containing one or more double bonds.

[0095] As used herein, the term "carbocyclylalkyl" refers to an alkyl group substituted with a carbocyclyl group.

[0096] The term "carbonate" is art-recognized and refers to a group: -OCO-R A (In the formula, R A represents a hydrocarbyl group).

[0097] As used herein, the term "carboxy" refers to a group represented by the formula: -CO2H.

[0098] As used herein, the term "ester" refers to the group: -C(O)OR A (In the formula, R A represents a hydrocarbyl group).

[0099] As used herein, the term "ether" refers to a hydrocarbyl group that is bonded to another hydrocarbyl group through an oxygen. Thus, the ether substituent of a hydrocarbyl group can be hydrocarbyl-O-. Ethers can be symmetrical or asymmetrical. Examples of ethers include, but are not limited to, heterocycle-O-heterocycle and aryl-O-heterocycle. Ethers include "alkoxyalkyl" groups, which can be represented by the general formula: alkyl-O-alkyl.

[0100] As used herein, the terms "halo" and "halogen" mean halogen and include chloro, fluoro, bromo, and iodo.

[0101] As used herein, the terms "hetaralkyl" and "heteroaralkyl" refer to an alkyl group substituted with a hetararyl group.

[0102] As used herein, the term "heteroalkyl" refers to a saturated or unsaturated chain of carbon atoms and at least one heteroatom, wherein no two heteroatoms are adjacent.

[0103] The terms "heteroaryl" and "hetaryl" include substituted or unsubstituted aromatic monocyclic ring structures containing at least one heteroatom, preferably 1 to 4 heteroatoms, more preferably 1 or 2 heteroatoms, and are preferably 5- to 7-membered, more preferably 5- to 6-membered. The terms "heteroaryl" and "hetaryl" also include polycyclic ring systems having two or more rings in which two or more carbon atoms are common to two adjacent rings, where at least one of the rings is heteroaromatic and the other rings may be, for example, cycloalkyl, cycloalkenyl, aryl, heteroaryl, and / or heterocyclyl. Heteroaryl groups include, for example, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, pyrazole, pyridine, pyrazine, pyridazine, and pyrimidine.

[0104] As used herein, the term "heteroatom" means an atom of any element other than carbon or hydrogen. Preferred heteroatoms are nitrogen, oxygen, and sulfur.

[0105] The terms "heterocyclyl," "heterocycle," and "heterocyclic" refer to a substituted or unsubstituted non-aromatic ring system containing at least one heteroatom, preferably 1 to 4 heteroatoms, more preferably 1 or 2 heteroatoms, and preferably a 3- to 10-membered ring, more preferably a 3- to 7-membered ring. The terms "heterocyclyl" and "heterocyclic" also include polycyclic ring systems having two or more rings in which two or more carbons are common to two adjacent rings, where at least one of the rings is heterocyclic, and the other rings, for example, can be cycloalkyl, cycloalkenyl, aryl, heteroaryl, and / or heterocyclyl. Heterocyclyl groups include, for example, piperidine, piperazine, pyrrolidine, tetrahydropyran, tetrahydrofuran, morpholine, lactones, lactams, and the like.

[0106] As used herein, the terms "heterocyclylalkyl" or "heterocycloalkyl" refer to an alkyl group substituted with a heterocyclyl group.

[0107] As used herein, the term "hydrocarbyl" refers to a group that does not have the =O or =S substituent, typically has at least one carbon-hydrogen bond, and a primarily carbon backbone, but may optionally contain heteroatoms, bonded through a carbon atom. Thus, groups such as methyl, ethoxyethyl, 2-pyridyl, and trifluoromethyl are considered hydrocarbyl for purposes of this invention, while substituents such as acetyl (having the =O substituent on the bonded carbon) and ethoxy (bonded through an oxygen rather than a carbon) are not considered hydrocarbyl. Hydrocarbyl groups include, but are not limited to, aryl, heteroaryl, carbocyclyl, heterocyclyl, alkyl, alkenyl, alkynyl, and combinations thereof.

[0108] As used herein, the term "hydroxyalkyl" refers to an alkyl group substituted with a hydroxy group.

[0109] The term "lower," when used in conjunction with chemical moieties such as acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy, is meant to include groups in which there are 10 or fewer, preferably 6 or fewer, non-hydrogen atoms present in the substituent. "Lower alkyl," for example, refers to alkyl groups containing 10 or fewer, preferably 6 or fewer, carbon atoms. In certain embodiments, an acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy substituent as defined herein is a lower acyl, lower acyloxy, lower alkyl, lower alkenyl, lower alkynyl, or lower alkoxy, respectively, whether occurring alone or in combination with other substituents such as, for example, hydroxyalkyl and aralkyl (in which case, for example, atoms in aryl groups are not counted when counting carbon atoms in an alkyl substituent).

[0110] The terms "polycyclyl," "polycycle," and "polycyclic" refer to two or more rings (e.g., cycloalkyl, cycloalkenyl, aryl, heteroaryl, and / or heterocyclyl) in which two or more atoms are common to two adjacent rings (e.g., the rings are "fused rings"). Each ring of a polycycle can be substituted or unsubstituted. In certain embodiments, each ring of a polycycle contains 3 to 10, preferably 5 to 7, atoms in the ring.

[0111] The term "silyl" refers to a silicon moiety having three hydrocarbyl moieties attached thereto.

[0112] The term "substituted" refers to moieties having substituents replacing a hydrogen on one or more backbone carbons. It is understood that "substituted" or "substituted with" includes the implicit proviso that such substitution results in a stable compound (e.g., does not undergo spontaneous transformation by rearrangement, cyclization, elimination, etc.), subject to the permissible valences of the substituted atom and substituent. As used herein, the term "substituted" is intended to include all permissible substituents of organic compounds. In a broad aspect, the permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and nonaromatic substituents of organic compounds. The permissible substituents can be one or more and the same or different for appropriate organic compounds. For purposes of this invention, heteroatoms, such as nitrogen, can have hydrogen substituents and / or any permissible substituents of organic compounds described herein that satisfy the valences of the heteroatoms. Substituents can include any of the substituents described herein, for example, halogen, hydroxyl, carbonyl (e.g., carboxyl, alkoxycarbonyl, formyl, or acyl), thiocarbonyl (e.g., thioester, thioacetate, or thioformate), alkoxy, phosphoryl, phosphate, phosphonate, phosphinate, amino, amido, amidine, imine, cyano, nitro, azido, sulfhydryl, alkylthio, sulfate, sulfonate, sulfamoyl, sulfonamido, sulfonyl, heterocyclyl, aralkyl, or aromatic or heteroaromatic moiety. In preferred embodiments, the substituent on the substituted alkyl is C 1~6 Alkyl, C 3~6In a more preferred embodiment, the substituent on the substituted alkyl is selected from fluoro, carbonyl, cyano, or hydroxyl. Those skilled in the art will understand that the substituents themselves may be substituted, where appropriate. Unless specifically stated as "unsubstituted," reference to a chemical moiety herein should be understood to include substituted variants. For example, reference to an "aryl" group or moiety implicitly includes both substituted and unsubstituted variants.

[0113] The term "sulfate" is art-recognized and refers to the group: -OSO3H, or a pharmaceutically acceptable salt thereof.

[0114] The term "sulfonamide" is art-recognized and refers to compounds having the general formula:

[0115] [ka] (wherein each R A independently represent hydrogen or hydrocarbyl (e.g., alkyl), or both R A together with the intervening atom(s) complete a heterocycle having 4 to 8 atoms in the ring structure).

[0116] The term "sulfoxide" is art-recognized and refers to the group: -S(O)-R A (In the formula, R A represents hydrocarbyl).

[0117] The term "sulfonate" is art-recognized and refers to the group SO3H, or a pharmaceutically acceptable salt thereof.

[0118] The term "sulfone" is art-recognized and refers to the group: -S(O)-R A (In the formula, R A represents hydrocarbyl).

[0119] As used herein, the term "thioalkyl" refers to an alkyl group substituted with a thiol group.

[0120] As used herein, the term "thioester" refers to the group: -C(O)SR A or -SC(O)R A (In the formula, R A represents hydrocarbyl).

[0121] As used herein, the term "thioether" corresponds to an ether where the oxygen has been replaced with a sulfur.

[0122] The term "urea" is art-recognized and has the general formula:

[0123] [ka] (wherein each R A independently represent hydrogen or hydrocarbyl (e.g., alkyl), or R A Any occurrence of, taken together with the other and the intervening atom(s), completes a heterocycle having from 4 to 8 atoms in the ring structure.

[0124] "Protecting group" refers to a group of atoms that, when attached to a reactive functional group in a molecule, masks, reduces, or prevents the reactivity of the functional group. Typically, a protecting group can be selectively removed as desired during synthesis. Examples of protecting groups are found in Greene and Wuts, Protective Groups in Organic Chemistry, 3 rdEd., 1999, John Wiley & Sons, NY, and Harrison et al., Compendium of Synthetic Organic Methods, Vols. 1-8, 1971-1996, John Wiley & Sons, NY Representative nitrogen protecting groups include, but are not limited to, formyl, acetyl, trifluoroacetyl, benzyl, benzyloxycarbonyl ("CBZ"), tert-butoxycarbonyl ("Boc"), trimethylsilyl ("TMS"), 2-trimethylsilyl-ethanesulfonyl ("TES"), trityl and substituted trityl groups, allyloxycarbonyl, 9-fluorenylmethyloxycarbonyl ("FMOC"), and nitro-veratryloxycarbonyl ("NVOC"). Representative hydroxyl protecting groups include, but are not limited to, those in which the hydroxyl group is acylated (esterified) or alkylated, such as benzyl and trityl ethers, as well as alkyl ethers, tetrahydropyranyl ethers, trialkylsilyl ethers (e.g., TMS or TIPS groups), glycol ethers, e.g., ethylene glycol and propylene glycol derivatives, and allyl ethers.

[0125] As used herein, a therapeutic agent that "prevents" or "reduces the risk of developing" a disease, disorder, or condition refers to a compound that, in a statistical sample, reduces the occurrence of the disease or condition in a treated sample compared to an untreated control sample, or delays the onset of or reduces the severity of one or more symptoms of the disease, disorder, or condition compared to an untreated control sample.

[0126] The term "treating" includes prophylactic and / or therapeutic treatment. The term "prophylactic or therapeutic treatment" is art-recognized and includes administration of one or more of the subject compositions to a host. If a therapeutic agent is administered prior to the onset of clinical signs of an undesired condition (e.g., a disease or other undesired condition in a host animal), the treatment is prophylactic (i.e., protects the host from the occurrence of the undesired condition). On the other hand, if administered after the appearance of an undesired condition, the treatment is therapeutic (i.e., is intended to reduce, ameliorate, or stabilize an existing undesired condition or its side effects).

[0127] The terms "co-administration" and "co-administered" refer to any form of administration of two or more different therapeutic compounds such that a second compound is administered while a previously administered therapeutic compound is still effective in the body (e.g., the two compounds are effective in the patient's body simultaneously, which may involve a synergistic effect of the two compounds). For example, the different therapeutic compounds can be administered simultaneously or sequentially, in the same formulation or in separate formulations. In certain embodiments, the different therapeutic compounds can be administered within 1 hour, 12 hours, 24 hours, 36 hours, 48 ​​hours, 72 hours, or 1 week of each other. Thus, an individual receiving such treatment can benefit from the combined effects of the different therapeutic compounds.

[0128] The term "prodrug" is intended to encompass compounds that are converted to the therapeutically active agents of the present invention under physiological conditions. One common method for producing prodrugs is to include one or more selected moieties that are hydrolyzed under physiological conditions to yield the desired molecule. In other embodiments, the prodrug is converted by the enzymatic activity of the host animal. For example, esters or carbonates (e.g., esters or carbonates of alcohols or carboxylic acids) are preferred prodrugs of the present invention. In certain embodiments, some or all of the compounds of the present invention in the formulations shown above may be replaced with the corresponding appropriate prodrug (e.g., providing a hydroxyl in the parent compound as an ester or carboxylate, or providing a carboxylic acid present in the parent compound as an ester).

[0129] As used herein, "small molecule" refers to an organic or inorganic molecule with a molecular weight below about 3,000 Daltons. Generally, small molecules useful for the present invention have a molecular weight below 3,000 Daltons (Da). The small molecule can be, for example, at least about 100 Da to about 3,000 Da (e.g., about 100 to about 3,000 Da, about 100 to about 2,500 Da, about 100 to about 2,000 Da, about 100 to about 1,750 Da, about 100 to about 1,500 Da, about 100 to about 1,250 Da, about 100 to about 1,000 Da, about 100 to about 750 Da, about 100 to about 500 Da, about 200 to about 1,500, about 500 to about 1,000, about 300 to about 1,000 Da, or about 100 to about 250 Da).

[0130] In some embodiments, "small molecule" refers to an organic, inorganic, or organometallic compound that typically has a molecular weight of less than about 1000. In some embodiments, small molecules are organic compounds on the order of 1 nm in size. In some embodiments, small molecule drugs of the present invention include oligopeptides and other biomolecules with a molecular weight of less than about 1000.

[0131] An "effective amount" is an amount sufficient to produce a beneficial or desired result. For example, a therapeutic amount is an amount that achieves a desired therapeutic effect. This amount may be the same as or different from a prophylactically effective amount, which is the amount needed to prevent the onset of a disease or a symptom of a disease. An effective amount can be administered in one or more administrations, applications, or dosages. The therapeutically effective amount of a composition depends on the composition selected. The composition can be administered once or more times daily to once or more times weekly (including once every other day). One of skill in the art will recognize that certain factors (including, but not limited to, the severity of the disease or disorder, previous treatments, the subject's overall health and / or age, and other diseases present) can affect the dosage and timing required to effectively treat a subject. Furthermore, treatment of a subject with a therapeutically effective amount of a composition described herein can include a single treatment or a series of treatments.

[0132] Compounds of the Invention One aspect of the present invention provides a method for treating renal disease, comprising co-administering to a subject in need thereof a TRPC5 inhibitory compound and a second therapeutic agent. In some embodiments, the TRPC5 inhibitory compound is a small molecule inhibitor of TRPC5.

[0133] Small molecule inhibitors of TRPC5 In some embodiments, the TRPC5 inhibitor compound is a compound of structural formula (A), or a tautomer or pharmaceutically acceptable salt thereof:

[0134] [ka] (In the formula, Each R is independently H, alkyl, alkenyl, alkynyl, aryl, heterocyclyl, heteroaryl, halogen, -OH, CN, cycloalkyl, -O-alkyl, -O-cycloalkyl, -O-aryl, -aryl-O-aryl, -CF3, -C(H)F2, alkylene-CF3, alkylene-C(H)F2, -SO2-alkyl, -O-alkylene-O-alkyl, -heterocyclyl-LR 4 , and heteroaryl-LR 4 selected from the group consisting of R 4 is absent or is an alkyl, cycloalkyl, polycyclyl, aryl, heterocyclyl, heteroaryl, -C(O)N(R 5 CF3, R 5 are independently H or alkyl; R 6 is an alkyl, cycloalkyl, aryl, heterocyclyl, heteroaryl, alkylene-aryl, -C(O)N(R 5 CF3, L is absent or is methylene, -C(O)-, -SO2-, -CH2N(Me)-, -N(R 5 )(R 6 )-, -C(R 5 )(R 6 )-, and -OR 6 selected from the group consisting of If only one R is -heterocyclyl-LR 4 or -heteroaryl-LR 4 (It is).

[0135] In some embodiments, the TRPC5 inhibitor compound is represented by structural formula (AI), (A-II), or (A-III), or a tautomer or pharmaceutically acceptable salt thereof:

[0136] [ka] (In the formula, R 1and R 3 are independently H, alkyl, alkenyl, alkynyl, aryl, heterocyclyl, heteroaryl, halogen, —OH, CN, cycloalkyl, —O-alkyl, —O-cycloalkyl, —O-aryl, -aryl-O-aryl, —CF, —C(H)F, alkylene-CF, alkylene-C(H)F, —SO-alkyl, and —O-alkylene-O-alkyl, -heterocyclyl-LR 4 , and heteroaryl-LR 4 selected from the group consisting of R 2 -heterocyclyl-LR 4 and R 4 is absent or is selected from alkyl, cycloalkyl, aryl, alkylene-aryl, alkylene-heteroaryl, heteroaryl, heterocyclyl, -C(O)N(R 5 CF3, R 5 are independently H or alkyl; R 6 is an alkyl, cycloalkyl, aryl, heterocyclyl, heteroaryl, alkylene-aryl, -C(O)N(R 5 CF3, L is absent or is selected from methylene, -C(O)-, -SO2-, -CH2N(Me)-, -N(R5)(R6)-, -C(R5)(R6)-, and -OR 6 selected from the group consisting of R 1 , R 2 , and R 3 and only one of the groups is -heterocyclyl-LR 4 or -heteroaryl-LR 4 (It is).

[0137] In some embodiments, the TRPC5 inhibitor compound is a compound disclosed in International Patent Application No. PCT / US18 / 51465, filed September 18, 2018, which is incorporated herein by reference in its entirety.

[0138] In some embodiments, the TRPC5 inhibitor compound is selected from any one of the following compounds, or a pharmaceutically acceptable salt thereof:

[0139] [Table 1-1]

[0140] [Table 1-2]

[0141] [Table 1-3]

[0142] In some embodiments, the TRPC5 inhibitor compound has structural formula (I):

[0143] [ka] or a pharmaceutically acceptable salt thereof (In the formula, "---" is a single or double bond, X 1 is CH or N, If "---" is a double bond, X 2 is CH or N, If "---" is a single bond, X 2 is N(CH3), X 1 If is CH, then X 2 is N or N(CH3), Y is -O-, -N(CH3)-, -N(CH2CH2OH)-, cyclopropane-1,1-diyl, or -CH(CH3)-; Q is 2-trifluoromethyl-4-fluorophenyl, 2-difluoromethyl-4-fluorophenyl, 2-trifluoromethylphenyl, 2-methyl-4-fluorophenyl, 2-chloro-4-fluorophenyl, 2-chlorophenyl, 1-(benzyl)-4-methylpiperidin-3-yl, 4-trifluoromethylpyridin-3-yl, 2-trifluoromethyl-6-fluorophenyl, 2-trifluoromethyl-3-cyanophenyl, 2-ethyl-3-fluorophenyl, 2-chloro-3-cyanophenyl, 2-trifluoromethyl-5-fluorophenyl, or 2-difluoromethylphenyl; If "---" is a double bond, R 13 is hydrogen, -CH2OH, -CH(OH)-CH2OH, -NH2, -CH(OH)CH3, -OCH3, or -NH-(CH2)2OH, and R 14 does not exist, or If "---" is a single bond, R 13 and R 14 together form =O, R 15 and R 16 each independently is hydrogen or —CH3. In some embodiments, X 1 N, X 2 is N, Y is —O— or —N(CH3)—, and Q is 2-trifluoromethylphenyl, then R 13 , R 15 , and R 16 At least one of these is not hydrogen.

[0144] In some embodiments, the TRPC5 inhibitor compound has structural formula (II):

[0145] [ka] or a pharmaceutically acceptable salt thereof, wherein R 11 is chloro, -CF3, -CHF2, or -CH3, R 12 is hydrogen or fluoro, R 13 is hydrogen, -NH2, -CH2OH, or CH(OH)-CH2OH).

[0146] In some embodiments, R 11 is -CHF2, and R 12 is fluoro.

[0147] In some embodiments, the TRPC5 inhibitor compound is selected from any one of the following compounds, or a pharmaceutically acceptable salt thereof:

[0148] [Table 2-1]

[0149] [Table 2-2]

[0150] [Table 2-3]

[0151] [Table 2-4]

[0152] [Table 2-5]

[0153] In some embodiments, the TRPC5 inhibitor compound is a compound disclosed in U.S. Provisional Patent Application No. 62 / 732,728, filed September 18, 2018, or U.S. Provisional Patent Application No. 62 / 780,553, filed December 17, 2018, each of which is incorporated by reference in its entirety.

[0154] In some embodiments, the TRPC5 inhibitor compound is selected from any one of the following compounds, or a pharmaceutically acceptable salt thereof:

[0155] [Table 3-1]

[0156] [Table 3-2]

[0157] In some embodiments, the TRPC5 inhibitor compound is selected from any one of the following compounds, or a pharmaceutically acceptable salt thereof:

[0158] [Table 4]

[0159] Second therapeutic agent In one aspect, the present invention is directed to a method of treating renal disease, comprising co-administering to a subject in need thereof a TRPC5 inhibitor compound and a second therapeutic agent. In certain embodiments, the second therapeutic agent affects a biological pathway other than the TRPC5-Rac1 pathway, and thus, the subject receiving such treatment can benefit from the combined effects of the different therapeutic agents.

[0160] In certain embodiments, the second therapeutic agent is selected from an immunomodulator, a calcineurin inhibitor, a renin-angiotensin-aldosterone system inhibitor, an antiproliferative agent, a corticosteroid, an angiotensin-converting enzyme inhibitor, an angiotensin receptor blocker, a sodium-glucose transport protein 2 inhibitor, a nuclear factor-1 (erythroid-derived 2)-like 2 agonist, a chemokine receptor 2 inhibitor, a chemokine receptor 5 inhibitor, and an endothelin 1 receptor antagonist.

[0161] In some embodiments, the second therapeutic agent is further selected from the following: an alkylating agent, an adrenocorticotropic hormone stimulator, a dual sodium-glucose transport protein 1 / 2 inhibitor, a beta blocker (e.g., metoprolol succinate, metoprolol tartrate, propranolol, carvedilol), a mineralocorticoid receptor antagonist (e.g., spironolactone, eplerenone, finerenone, esaxerenone, aparenone), a loop or thiazide diuretic (e.g., furosemide, bumetanide, or bendroflumethiazide), a calcium channel blocker (verapamil, diltiazem, amlodipine, or nifedipine), a statin (e.g., atorvastatin, pravastatin, fluvastatin, lovastatin, rosuvastatin, simvastatin, or pitavastatin), a short-, intermediate-, or long-acting insulin (e.g., NPH insulin (Humulin R, Novolin R, biosimilars), insulin lispro (Humalog), insulin glulisine), insulin glargine (Basaglar, Lantus), insulin detemir (Levemir), insulin degludec (Tresiba)), dipeptidyl peptidase 4 inhibitors (e.g., sitagliptin, saxagliptin, linagliptin, vildagliptin), glucagon-like peptide 1 receptor agonists (e.g., exenatide, liraglutide, dulaglutide, lixisenatide, albiglutide, semaglutide), sulfonylureas (e.g., glimepiride, glipizide, glyburide, glibenclamide, chlorpropamide) , tolazamide, or tolbutamide), apoptosis signal-regulating kinase-1 (e.g., selonsertib), chymase inhibitors (e.g., furasimustat (BAY1142524), selective glycation inhibitors (e.g., GLY-230), renin inhibitors (e.g., SCO-272), interleukin-33 inhibitors (e.g., MEDI-3506), farnesoid X receptor agonists (e.g., nidufexor (LMB763)), soluble guanylate cyclase stimulators (e.g., praliciguat, olinciguat, IW-6463, vericiguat, riociguat), thromboxane receptor antagonists (e.g., SER150),Xanthine oxidase inhibitors (TMX-049), erythropoietin receptor agonists (sibinetide (ARA-290)), cannabinoid receptor type 1 inverse agonists (e.g., nimasimab, GFB-024, CRB-4001), NADPH oxidase inhibitors (e.g., APX-115), anti-vascular endothelial growth factor B (e.g., CSL-346), anti-fibrotic agents (e.g., FT011), neprilysin inhibitors (e.g., TD-1439, TD-0714, sacubitril), dual CD80 / CD86 inhibitors (e.g., abatacept), CD40 antagonists (e.g., bleselumab (ASKP1240)), cellular cholesterol and lipid blockers (VAR-200), PDGFR antagonists (e.g., ANG_3070) , Slit guidance ligand 2 (e.g., PF-06730512), APOL1 inhibitors (e.g., VX-147), Nrl2 activators / NF-κB inhibitors (e.g., bardoxolone), somatostatin receptor agonists (e.g., lanreotide), PPAR gamma agonists (e.g., pioglitazone), AMP-activated protein kinase stimulators (e.g., metformin), tyrosine kinase inhibitors (e.g., tecevatinib), glucosylceramide synthase inhibitors (e.g., benglustat malate), arginine vasopressin receptor 2 antagonists (e.g., lixivaptan), xanthine oxidase inhibitors (e.g., oxypurinol), or vasopressin receptor 2 antagonists (e.g., tolvaptan).

[0162] In some embodiments, the immunomodulator is rituximab. Rituximab destroys both normal and malignant B cells that have CD20 on their surface and is therefore used to treat diseases characterized by having excess, overactive, or dysfunctional B cells. Such diseases include, but are not limited to, hematological cancers and autoimmune diseases.

[0163] In some embodiments, the immunomodulator is mycophenolate mofetil, the administration of which can confer beneficial effects such as suppressing the immune system and preventing rejection during organ transplants.

[0164] In some embodiments, the angiotensin-converting enzyme inhibitor is captopril, zofenopril, enalapril, ramipril, quinapril, perindopril, lisinopril, benazepril, imidapril, trandolapril, or cilazapril. Angiotensin-converting enzyme (ACE) inhibitors are primarily used to treat hypertension and congestive heart failure. This class of drugs relaxes blood vessels and reduces blood volume, thereby lowering blood pressure and reducing oxygen demand from the heart. This class of drugs inhibits angiotensin-converting enzyme, a key component of the renin-angiotensin system. They are also useful in treating other cardiovascular and renal diseases, including, but not limited to, acute myocardial infarction (heart attack), heart failure (left ventricular systolic dysfunction), and renal complications of diabetes (diabetic nephropathy).

[0165] In some embodiments, the angiotensin receptor blocker is losartan, candesartan, valsartan, irbesartan, telmisartan, eprosartan, olmesartan, azilsartan, or fimasartan. Uses of angiotensin receptor blockers include, but are not limited to, the treatment of elevated blood pressure (high blood pressure), diabetic nephropathy (kidney damage caused by diabetes), and congestive heart failure.

[0166] In some embodiments, the renin-angiotensin-aldosterone system inhibitor is aliskiren. Inhibition of the renin-angiotensin-aldosterone system can confer beneficial effects such as lowering blood pressure and improving intraglomerular hemodynamics. Renin is the first enzyme in the renin-angiotensin-aldosterone system and is responsible for blood pressure control. Renin cleaves angiotensinogen to angiotensin I, which is then converted to angiotensin II by angiotensin-converting enzyme (ACE). Angiotensin II has direct and indirect effects on blood pressure. Angiotensin II directly constricts arterial smooth muscle, causing vasoconstriction and elevated blood pressure. Angiotensin II also stimulates the production of aldosterone from the adrenal cortex, which increases renal tubular sodium reabsorption, followed by water, thereby increasing plasma volume and blood pressure. Aliskiren binds to the S3bp binding site of renin, which is essential for renin activity. Binding to this pocket inhibits the conversion of angiotensinogen to angiotensin I. Aliskiren is also available as a combination therapy with hydrochlorothiazide.

[0167] In some embodiments, the endothelin 1 receptor antagonist is ambrisentan, atrasentan, bosentan, macitentan, or sparsentan. Endothelin 1 receptor antagonism can have beneficial effects such as lowering blood pressure and improving intraglomerular hemodynamics. Macitentan, ambrisentan, and bosentan are primarily used to treat pulmonary arterial hypertension, which may have a multifactorial mechanism and may include chronic renal failure.

[0168] In some embodiments, the antiproliferative agent is mycophenolate mofetil, mycophenolate sodium, or azathioprine. Administration of mycophenolate mofetil, mycophenolate sodium, or azathioprine can confer beneficial effects such as suppressing the immune system and preventing rejection in organ transplants.

[0169] In some embodiments, the SGLT2 inhibitor is canagliflozin, dapagliflozin, empagliflozin, a combination of empagliflozin and linagliptin, a combination of empagliflozin and metformin, or a combination of dapagliflozin and metformin. Inhibition of SGLT2 can confer beneficial effects such as glucose lowering and improved intraglomerular hemodynamics. SGLT2 inhibitors (also called gliflozins) are a class of medications that lower blood glucose levels by inhibiting glucose reabsorption in the kidney. They act by inhibiting sodium-glucose transport protein 2 (SGLT2). SGLT2 inhibitors are used to treat type 2 diabetes mellitus (T2DM). Aside from glycemic control, gliflozins have been shown to provide significant cardiovascular benefits to T2DM patients. Studies of canagliflozin have shown that the medication not only promotes glycemic control but also reduces weight and systolic and diastolic blood pressure. Sodium-glucose cotransporters (SGLTs) are proteins found primarily in the kidney and play an important role in maintaining blood glucose balance. SGLT1 and SGLT2 are the two most well-known SGLTs in this family. SGLT2 is the primary transporter, facilitating the reabsorption of glomerular-filtered glucose into the circulating blood, accounting for approximately 90% of renal glucose reabsorption. SGLT2 is primarily expressed on epithelial cells lining the first segment of the proximal convoluted tubule in the kidney. Gliflozin inhibits SGLT2, thereby inhibiting the kidney's reuptake of glucose from the glomerular filtrate, subsequently reducing glucose levels and promoting urinary glucose excretion (glycosuria).

[0170] In some embodiments, the SGLT2 inhibitor also inhibits SGLT1. In some aspects of these embodiments, the SGLT1 / 2 inhibitor is sotagliflozin.

[0171] In some embodiments, the calcineurin inhibitor is cyclosporine A, voclosporin, or tacrolimus. Calcineurin (CaN) is a calcium-calmodulin-dependent serine / threonine protein phosphatase (also known as protein phosphatase 3-calcium-dependent serine / threonine phosphatase). Calcineurin activates T cells of the immune system and can be blocked by drugs (including, but not limited to, cyclosporine, voclosporin, pimecrolimus, and tacrolimus). Calcineurin activates the transcription factor cytoplasmic nuclear factor of activated T cells (NFATc) by dephosphorylating it. Activated NFATc then translocates to the nucleus, where it upregulates interleukin-2 (IL-2) expression and stimulates the growth and differentiation of T cell responses. Calcineurin inhibitors, such as tacrolimus, are used to suppress the immune system of organ allograft recipients and prevent rejection of the transplanted tissue.

[0172] In some embodiments, the nuclear factor-1 (erythroid-derived 2)-like 2 agonist is bardoxolone or CXA-10. Nuclear factor-1 (erythroid-derived 2)-like 2 agonism can confer beneficial effects, such as anti-inflammatory effects. Nuclear factor (erythroid-derived 2)-like 2, also known as NFE2L2 or Nrf2, is a transcription factor encoded by the NFE2L2 gene in humans. Nrf2 is a basic leucine zipper (bZIP) protein that regulates the expression of antioxidant proteins that protect against oxidative damage caused by injury and inflammation. Several drugs that stimulate the NFE2L2 pathway are being investigated for the treatment of diseases caused by oxidative stress. Heme oxygenase-1 (HMOX1, HO-1) is an enzyme that breaks down heme into the antioxidant biliverdin, the anti-inflammatory carbon monoxide, and iron. HO-1 is an Nrf2 target gene and has been shown to protect against a variety of pathologies, including sepsis, elevated blood pressure, atherosclerosis, acute lung injury, kidney damage, and pain.

[0173] In some embodiments, the chemokine receptor 2 inhibitor is PF-04136309, ccx140, or propagermanium (DMX-200). Inhibition of chemokine receptor 2 can have beneficial effects, such as suppression of the immune system. Recruitment of monocytes and other inflammatory cells via chemokine receptor 2 (CCR2) is involved in the pathogenesis of diabetic nephropathy, and inhibiting CCR2 may reduce albuminuria and prevent the decline of renal function in patients with diabetic nephropathy.

[0174] In some embodiments, the second therapeutic agent is an Nrl2 activator / NF-κB inhibitor (e.g., bardoxolone), a somatostatin receptor agonist (e.g., lanreotide), a PPAR gamma agonist (e.g., pioglitazone), an AMP-activated protein kinase stimulator (e.g., metformin), a tyrosine kinase inhibitor (e.g., tecevatinib), a glucosylceramide synthase inhibitor (e.g., benglustat malate), an arginine vasopressin receptor 2 antagonist (e.g., lixivaptan), a xanthine oxidase inhibitor (e.g., oxypurinol), or a vasopressin receptor 2 antagonist (e.g., tolvaptan). Each of these agents is approved for the treatment of polycystic kidney disease, particularly autosomal dominant polycystic kidney disease, or is undergoing human clinical trials.

[0175] In some embodiments, the second therapeutic agent is tacrolimus, cyclosporin A, rituximab, mycophenolate mofetil, corticosteroids (e.g., prednisone), sparsentan, enalapril, or losartan. In some embodiments, the second therapeutic agent is voclosporin. In some embodiments, the second therapeutic agent is enalapril, losartan, or cyclosporin A. Corticosteroids are a class of steroid hormones produced in the adrenal cortex of vertebrates, as well as synthetic analogs of these hormones. The two major classes of corticosteroids, glucocorticoids and mineralocorticoids, are involved in a wide range of physiological processes, including stress response, immune response, and regulation of inflammation, carbohydrate metabolism, protein catabolism, blood electrolyte levels, and behavior. Mineralocorticoids, such as aldosterone, are primarily involved in regulating electrolyte and water balance by regulating ion transport in renal tubular epithelial cells. Systemic corticosteroids are also used to treat diseases and conditions such as nephrotic syndrome, organ transplantation, adrenal insufficiency, and congenital adrenal hyperplasia.

[0176] In certain embodiments, the compound of the present invention can be racemic.In certain embodiments, the compound of the present invention can be enriched in one enantiomer.For example, the compound of the present invention can have an enantiomeric excess (EE) of more than 30%, more than 40%, more than 50%, more than 60%, more than 70%, more than 80%, more than 90%, or even more than 95%.

[0177] The compounds of the present invention may have multiple stereocenters. Thus, the compounds of the present invention may be enriched with one or more diastereomers. For example, the compounds of the present invention may have more than 30% de (diastereomeric excess), more than 40% de, more than 50% de, more than 60% de, more than 70% de, more than 80% de, more than 90% de, or even more than 95% de. In certain embodiments, the compounds of the present invention have substantially one isomeric configuration at one or more asymmetric centers, and multiple isomeric configurations at the remaining asymmetric centers.

[0178] In certain embodiments, the enantiomeric excess of the asymmetric center is at least 40% ee, 50% ee, 60% ee, 70% ee, 80% ee, 90% ee, 92% ee, 94% ee, 95% ee, 96% ee, 98% ee, or higher ee.

[0179] As used herein, a single bond drawn without stereochemistry does not indicate the stereochemistry of the compound.

[0180] As used herein, dashed or bold non-wedged bonds indicate relative but not absolute stereochemical configurations (e.g., do not distinguish between enantiomers of a given diastereomer).

[0181] As used herein, a dashed or bold wedge bond denotes absolute stereochemical configuration.

[0182] In some embodiments, the present invention relates to pharmaceutical compositions comprising a compound of the present invention and a pharmaceutically acceptable carrier. In certain embodiments, therapeutic preparations or pharmaceutical compositions of the present invention may be predominantly enriched in one enantiomer of a compound. An enantiomerically enriched mixture may contain, for example, at least 60 mole percent, or more preferably at least 75, 90, 95, or even 99 mole percent of one enantiomer. In certain embodiments, a compound enriched in one enantiomer is substantially free of the other enantiomer. In this context, substantially free means, for example, that the substance constitutes less than 10%, or less than 5%, or less than 4%, or less than 3%, or less than 2%, or less than 1% of the other enantiomer in the composition or compound mixture. For example, a composition or compound mixture containing 98 grams of a first enantiomer and 2 grams of a second enantiomer would be considered to contain 98 mole percent of the first enantiomer and only 2% of the second enantiomer.

[0183] In certain embodiments, therapeutic preparations or pharmaceutical compositions may be enriched to provide predominantly one diastereomer of a compound of the invention. A diastereomerically enriched mixture can contain, for example, at least 60 mole percent, or more preferably at least 75, 90, 95, or even 99 mole percent of one diastereomer.

[0184] Treatment method Non-selective Ca 2+Transient receptor potential (TRP) channels function as sensors that transmit extracellular cues to the intracellular environment in diverse cellular processes, including actin remodeling and cell migration (Greka et al., Nat Neurosci 6, 837-845, 2003; Ramsey et al., Annu Rev Physiol 68, 619-647, 2006; Montell, Pflugers Arch 451, 19-28, 2005; Clapham, Nature 426, 517-524, 2003). Dynamic reorganization of the actin cytoskeleton is a key component of spatiotemporally regulated Ca ions. 2+ These changes depend on the influx of ATP (Zheng and Poo, Annu Rev Cell Dev Biol 23, 375-404, 2007; Brandman and Meyer, Science 322, 390-395, 2008; Collins and Meyer, Dev Cell 16, 160-161, 2009), and the small GTPases RhoA and Rac1 function as key modulators of these changes (Etienne-Manneville and Hall, Nature 420, 629-635, 2002; Raftopoulou and Hall, Dev Biol 265, 23-32, 2004). RhoA induces stress fiber and focal adhesion formation, while Rac1 mediates lamellipodia formation (Etienne-Manneville and Hall, Nature 420, 629-635, 2002). Transient receptor potential cation channel subfamily C member 5 (TRPC5) acts in concert with TRPC6 to regulate Ca2+ influx, actin remodeling, and cell motility in kidney podocytes and fibroblasts. TRPC5-mediated Ca 2+TRPC6-mediated Ca2+ influx promotes RhoA activity, whereas TRPC6 channel gene silencing eliminates stress fibers, reduces punctate contacts, and leads to a motile, migratory cellular phenotype. In contrast, TRPC5 channel gene silencing rescues stress fiber formation and leads to a contractile cellular phenotype. The results described herein reveal a conserved signaling mechanism by which TRPC5 and TRPC6 channels control a tightly regulated balance of cytoskeletal dynamics through differential binding to Rac1 and RhoA.

[0185] Actin cytoskeleton Ca 2+ Ca-dependent remodeling is a dynamic process that promotes cell migration (Wei et al., Nature 457, 901-905, 2009). RhoA and Rac1 function as switches involved in cytoskeletal reorganization in migrating cells (Etienne-Manneville and Hall, Nature 420, 629-635, 2002; Raftopoulou and Hall, Dev Biol 265, 23-32, 2004). Rac1 activation mediates a motile cell phenotype, while RhoA activity promotes a contractile phenotype (Etienne-Manneville and Hall, Nature 420, 629-635, 2002). 2+ Ca plays a central role in the regulation of small GTPases (Aspenstrom et al., Biochem J 377, 327-337, 2004). 2+Spatially and temporally restricted flickering of Rac1 is concentrated near the leading edge of migrating cells (Wei et al., Nature 457, 901-905, 2009). Thus, Ca2+ microdomains coordinate local bursts of Rac1 activity as a key event at the leading edge (Gardiner et al., Curr Biol 12, 2029-2034, 2002; Machacek et al., Nature 461, 99-103, 2009). So far, the source of Ca2+ influx involved in GTPase regulation remains largely unknown. TRP (Transient Receptor Potential) channels are known to mediate the temporally and spatially restricted Ca2+ influx linked to cell migration in fibroblast and neuronal growth cones. 2+ Specifically, TRPC5 channels are known regulators of neuronal growth cone guidance1, and their activity in neurons is dependent on PI3K and Rac1 activity (Bezzerides et al., Nat Cell Biol 6, 709-720, 2004).

[0186] Podocytes are neuron-like cells derived from the metanephric mesenchyme of the kidney glomerulus and are essential for the formation of the kidney filtration apparatus (Somlo and Mundel, Nat Genet. 24, 333-335, 2000; Fukasawa et al., J Am Soc Nephrol 20, 1491-1503, 2009). Podocytes have an exquisitely elaborated repertoire of cytoskeletal adaptations to environmental cues (Somlo and Mundel, Nat Genet 24, 333-335, 2000; Garg et al., Mol Cell Biol 27, 8698-8712, 2007; Verma et al., J Clin Invest 116, 1346-1359, 2006; Verma et al., J Biol Chem 278, 20716-20723, 2003; Barletta et al., J Biol Chem 278, 19266-19271, 2003; Holzman et al., Kidney Int 56, 1481-1491, 1999; Ahola et al., Am J Pathol 155,907-913,1999;Tryggvason and Wartiovaara,N Engl J Med 354,1387-1401,2006;Schnabel and Farquhar,J Cell Biol 111,1255-1263,1990;Kurihara et al.,Proc Natl Acad Sci USA 89,7075-7079,1992). Early events in podocyte injury are characterized by dysregulation of the actin cytoskeleton (Faul et al., Trends Cell Biol 17, 428-437, 2007; Takeda et al., J Clin Invest 108, 289-301, 2001; Asanuma et al., Nat Cell Biol 8, 485-491, 2006) and Ca2+ homeostasis (Hunt et al., J Am Soc Nephrol 16, 1593-1602, 2005; Faul et al., Nat Med 14, 931-938, 2008).These changes are associated with the development of proteinuria, loss of albumin into the urinary space, and ultimately renal failure (Tryggvason and Wartiovaara, N Engl J Med 354, 1387-1401, 2006). The vasoactive hormone angiotensin II stimulates podocyte Ca2+ levels. 2+ Treatment with ATP induces Ca2+ influx, and long-term treatment results in the loss of stress fibers (Hsu et al., J Mol Med 86, 1379-1394, 2008). Although the connection between Ca2+ influx and cytoskeletal reorganization is recognized, the mechanisms by which podocytes sense and transduce extracellular cues that regulate cell shape and motility remain unclear. Mutations in the TRP canonical 6 (TRPC6) channel have been linked to podocyte injury (Winn et al., Science 308, 1801-1804, 2005; Reiser et al., Nat Genet 37, 739-744, 2005; Moller et al., J Am Soc Nephrol 18, 29-36, 2007; Hsu et al., Biochim Biophys Acta 1772, 928-936, 2007), but the specific pathways regulating this process remain largely unknown. Furthermore, TRPC6 shares close similarity with the other six members of the TRPC channel family (Ramsey et al., Annu Rev Physiol 68, 619-647, 2006; Clapham, Nature 426, 517-524, 2003). TRPC5 channels antagonize TRPC6 channel activity and control a tightly regulated balance of cytoskeletal dynamics through differential binding to different small GTPases.

[0187] Proteinuria Proteinuria is a pathological condition characterized by the presence of protein in the urine. Albuminuria is a type of proteinuria. Microalbuminuria occurs when the kidneys leak small amounts of albumin into the urine. In a properly functioning body, albumin is normally absent from the urine because it is retained in the bloodstream by the kidneys. Microalbuminuria is diagnosed based on a 24-hour urine collection (20-200 μg / min) or, more commonly, at least two elevated concentrations (30-300 mg / L). Microalbuminuria can be a precursor to diabetic nephropathy. Albumin levels higher than these values ​​are called macroalbuminuria. For example, subjects with certain diseases (e.g., diabetic nephropathy) progress from microalbuminuria to macroalbuminuria, reaching the nephropathic range (>3.5 g / 24 hours) as kidney disease progresses.

[0188] Causes of Proteinuria Proteinuria can be associated with several conditions, including focal segmental glomerulosclerosis, IgA nephropathy, diabetic nephropathy, lupus nephritis, membranoproliferative glomerulonephritis, progressive (crescentic) glomerulonephritis, and membranous glomerulonephritis.

[0189] A. Focal segmental glomerulosclerosis (FSGS) Focal segmental glomerulosclerosis (FSGS) is a disease that attacks the kidney's filtration system (glomeruli), causing severe scarring. FSGS is one of many causes of a condition known as nephrotic syndrome, which occurs when blood proteins leak into the urine (proteinuria). Primary FSGS usually presents as nephrotic syndrome when no underlying disease cause is identified. Secondary FSGS usually presents as kidney failure and proteinuria when an underlying disease cause is identified. FSGS can be hereditary, and several known genetic causes of hereditary forms of FSGS are currently recognized.

[0190] There are very few treatments available for patients with FSGS. Many patients are treated with steroid regimens, many of which are associated with severe side effects. Some patients have been shown to respond positively to immunosuppressants as well as blood pressure medications, and these medications have been shown to reduce urinary protein levels. To date, there is no universally accepted, effective treatment or therapy, and no FDA-approved medications exist to treat FSGS. Therefore, more effective methods for reducing or inhibiting proteinuria are desirable.

[0191] B. IgA nephropathy IgA nephropathy (also known as IgA nephritis, IgAN, Berger's disease, or glomerulonephritis with pharyngitis) is a form of glomerulonephritis (inflammation of the kidney's glomeruli). IgA nephropathy is the most common form of glomerulonephritis worldwide. Primary IgA nephropathy is characterized by the deposition of IgA antibodies in the glomeruli. There are other diseases associated with glomerular IgA deposition, the most common of which is Henoch-Schönlein purpura (HSP), which is considered a systemic form of IgA nephropathy. Henoch-Schönlein purpura presents with a characteristic purpuric skin rash, arthritis, and abdominal pain and occurs more commonly in young adults (ages 16–35). HSP has a more benign prognosis than IgA nephropathy. IgA nephropathy slowly progresses to chronic renal failure over a 20-year period in 25–30% of cases.

[0192] C. Diabetic nephropathy Diabetic nephropathy, also known as Kimmelstell-Wilson syndrome and intracapillary glomerulonephritis, is a progressive kidney disease caused by angiopathy of the glomerular capillaries. It is characterized by nephrotic syndrome and diffuse glomerular sclerosis. Diabetic nephropathy results from long-standing diabetes and is the primary cause of dialysis. The earliest detectable change in the course of diabetic nephropathy is thickening of the glomeruli. At this stage, the kidneys may begin to leak more serum albumin than normal into the urine. As diabetic nephropathy progresses, an increasing number of glomeruli are destroyed by nodular glomerulosclerosis, leading to increased amounts of albumin excreted in the urine.

[0193] D. Lupus nephritis Lupus nephritis is a kidney disorder that is a complication of systemic lupus erythematosus (SLE). It occurs when antibodies and complement build up in the kidneys, causing inflammation. Lupus nephritis often causes proteinuria and can rapidly progress to kidney failure. Nitrogenous waste products build up in the bloodstream. SLE causes various damage to the internal structures of the kidneys, including interstitial nephritis. Lupus nephritis affects approximately 3 in 10,000 people.

[0194] E. Membranoproliferative glomerulonephritis I / II / III Membranoproliferative glomerulonephritis (MPN) is a type of glomerulonephritis caused by the deposition of mesangial deposits and thickening of the basement membrane in the renal glomeruli, which activates complement and damages the glomeruli. There are three types of MNPN: Type I, which is caused by immune complex deposition in the kidney and is thought to be related to the classical complement pathway; Type II, which is similar to MNPN but is thought to be related to the alternative complement pathway; and Type III, which is extremely rare and is characterized by a combination of subepithelial deposits and the typical pathology of Type I disease.

[0195] There are two major types of MPGN based on immunofluorescence microscopy: immune complex-mediated and complement-mediated. Hypocomplementemia is common in all types of MPGN. In immune complex-mediated MPGN, complement activation occurs via the classical pathway and typically manifests as normal or mildly low serum C3 and C4 concentrations. Complement-mediated MPGN usually has low serum C3 and normal C4 levels due to activation of the alternative pathway. However, a normal serum C3 concentration does not exclude complement-mediated MPGN; therefore, normal C3 concentrations are not uncommon in adults with dense deposit disease (DDD) or C3 glomerulonephritis (C3GN).

[0196] C3 glomerulonephritis (C3GN) shows glomerulonephritis by light microscopy (LM), bright C3 staining by immunofluorescence microscopy (IF) but no C1q, C4, or immunoglobulin (Ig) staining, and mesangial and / or subendothelial electron-dense deposits by electron microscopy (EM). Scattered intramembranous and subepithelial deposits are also common. The term "C3 glomerulonephropathy" is often used to encompass C3GN and dense deposit disease (DDD), both of which result from dysregulation of the alternative complement pathway (AP). C3GN and DDD are difficult to distinguish from each other when studied by LM and IF. However, EM reveals mesangial and / or subendothelial, intramembranous, and subepithelial deposits in C3GN, while dense osmiophilic deposits in the glomerular basement membrane (GBM) and mesangium in DDD. Both C3GN and DDD are distinguished from immune complex-mediated glomerulonephritis by the lack of immunoglobulin staining on IF (Sethi et al., Kidney Int. (2012) 82(4):465-473).

[0197] F. Progressive (crescentic) glomerulonephritis Progressive (crescentic) glomerulonephritis (PG) is a renal syndrome that, if left untreated, rapidly progresses to acute renal failure and death within several months. PG is associated with an underlying condition, such as Goodpasture's syndrome, systemic lupus erythematosus, or Wegener's granulomatosis, in 50% of cases; the remaining cases are idiopathic. Regardless of the underlying cause, PG involves significant damage to the renal glomeruli, many of which have characteristic crescent-shaped scars. Patients with PG have hematuria, proteinuria, and, occasionally, elevated blood pressure and edema. Although the clinical picture is consistent with nephrotic syndrome, the degree of proteinuria can exceed 3 g / 24 hours, the range associated with nephrotic syndrome. If the disease is untreated, it may progress to decreased urine output (oliguria) as renal function declines.

[0198] G. Membranous glomerulonephritis Membranous glomerulonephritis (MGN) is a slowly progressive kidney disease that primarily affects patients between the ages of 30 and 50, usually Caucasians. Membranous glomerulonephritis can progress to nephrotic syndrome. MGN is caused by circulating immune complexes. Current research indicates that the majority of immune complexes are formed by in situ binding of antibodies to antigens on the glomerular basement membrane. These antigens may be endogenous to the basement membrane or deposited from the systemic circulation.

[0199] H. Alport syndrome Alport syndrome is a genetic disorder that affects approximately 1 in 5,000 to 10,000 children and is characterized by glomerulonephritis, end-stage renal disease, and hearing loss. Alport syndrome can also affect the eyes, but vision is usually not affected except in later cases when changes to the lens occur. Hematuria is common. Proteinuria is one of the features that develops as kidney disease progresses.

[0200] I. Hypertensive kidney disease Hypertensive kidney disease (hypertensive nephrosclerosis (HN) or HNS) or hypertensive nephropathy (HN)) is a medical condition that refers to damage to the kidneys caused by chronic hypertension. HN is classified into two types: benign and malignant. Benign nephrosclerosis is common in people over 60 years of age, while malignant nephrosclerosis is rare, affecting 1–5% of hypertensive patients with diastolic blood pressure above 130 mmHg. Signs and symptoms of chronic kidney disease (including loss of appetite, nausea, vomiting, pruritus, drowsiness or confusion, weight loss, and an unpleasant taste in the mouth) may occur. Chronic hypertension causes damage to kidney tissue, including small blood vessels, glomeruli, renal tubules, and interstitial tissue. The tissue hardens and thickens (this is known as nephrosclerosis). Narrowing of blood vessels means that less blood is delivered to the tissues, and therefore less oxygen reaches them, resulting in tissue death (ischemia).

[0201] J. Nephrotic syndrome Nephrotic syndrome is a group of symptoms resulting from kidney damage. Nephrotic syndrome includes proteinuria, low blood albumin levels, high blood lipids, and significant swelling. Other symptoms can include weight gain, fatigue, and foamy urine. Complications can include blood clots, infections, and high blood pressure. Causes include multiple kidney diseases, such as focal segmental glomerulosclerosis, membranous nephropathy, and minimal change nephrotic syndrome. It can also occur as a complication of diabetes or lupus. The underlying mechanism typically involves damage to the kidney's glomeruli. Diagnosis is typically based on urinalysis and, occasionally, kidney biopsy. Nephrotic syndrome differs from nephritic syndrome in that red blood cells are not present in the urine. Nephrotic syndrome is characterized by massive proteinuria (more than 3.5 g per 1.73 m2 of body surface area per day, or more than 40 mg per square meter of body surface area per hour in children), hypoalbuminemia (less than 2.5 g / dL), hyperlipidemia, and edema, beginning with the face. Lipiduria (lipids in the urine) may also occur but is not required for the diagnosis of nephrotic syndrome. Hyponatremia also occurs when sodium excretion is low. Genetic forms of nephrotic syndrome are typically resistant to steroids and other immunosuppressive treatments. The goals of treatment are to control urinary protein loss and swelling, provide adequate nutrition to allow the child to grow, and prevent complications. Early and aggressive treatment is used to control the disorder.

[0202] K. Minimal change nephrotic syndrome Minimal change nephrotic syndrome (MCS), also known as minimal change glomerulopathy (MCG), or Nir disease, is a disease that affects the kidneys and causes nephrotic syndrome. Clinical signs of minimal change nephrotic syndrome include proteinuria (abnormal urinary excretion of protein, primarily albumin), edema (swelling of soft tissues as a result of fluid retention), weight gain, and hypoalbuminemia (low serum albumin). Collectively, these signs are referred to as nephrotic syndrome. The first clinical sign of minimal change nephrotic syndrome is usually edema and associated weight gain. While swelling can be mild, patients may also present with lower extremity edema, periorbital edema, and swelling in the scrotal / labial area, and in severe cases, anasarca. In elderly patients, patients may also present with acute kidney injury (20–25% of affected adults) and hypertension. Due to the disease process, patients with minimal change nephrotic syndrome are also at risk for blood clots and infections.

[0203] L.Membranous nephropathy Membranous nephropathy refers to the deposition of immune complexes on the glomerular basement membrane (GBM), accompanied by thickening of the GBM. The cause is usually unknown (idiopathic), but secondary causes include drugs, infections, autoimmune disorders, and cancer. Findings include insidious onset of edema and heavy proteinuria with benign urinary sediment, normal renal function, and normal or elevated blood pressure. Membranous nephropathy is diagnosed by kidney biopsy. Spontaneous remission is common. Treatment for patients at high risk of progression typically involves corticosteroids and cyclophosphamide or chlorambucil.

[0204] M. postinfectious glomerulonephritis Acute proliferative glomerulonephritis is damage to the glomeruli (glomerulonephritis), i.e., the small blood vessels of the kidney. It is a common complication of bacterial infection, typically skin infections caused by Streptococcus types 12, 4, and 1 (impetigo), but can also occur after streptococcal pharyngitis, also known as postinfectious or poststreptococcal glomerulonephritis. This nephritis can be a risk factor for future albuminuria. In adults, signs and symptoms of infection may still be present at the time of kidney problems, and the terms infection-associated glomerulonephritis or bacterial infection-associated glomerulonephritis are also used. Deaths from acute glomerulonephritis worldwide have decreased from 24,000 in 1990 to 19,000 in 2013. Acute proliferative glomerulonephritis (poststreptococcal glomerulonephritis) occurs when streptococci infect the kidney (usually in the throat or skin), usually three weeks after infection, allowing time for antibodies and complement proteins to be produced. The infection causes inflammation of the kidney's blood vessels, which interferes with the kidney's ability to filter urine.[citation needed] Acute proliferative glomerulonephritis is most commonly seen in children.

[0205] N. Thin basement membrane disease Thin basement membrane disease (TBMD, also known as benign familial hematuria and thin basement membrane nephropathy (TBMN)), along with IgA nephropathy, is the most common cause of asymptomatic hematuria. The only abnormal finding in this disease is thinning of the glomerular basement membrane of the kidney. Its significance lies in the fact that patients maintain normal renal function throughout their lives and have a favorable prognosis. Most patients with thin basement membrane disease have incidental microscopic hematuria detected on urinalysis. Blood pressure, renal function, and urinary protein excretion are usually normal, although a small percentage of patients have mild proteinuria (<1.5 g / day) and elevated blood pressure. The presence of frank hematuria and lumbar pain should prompt investigation for other causes, such as kidney stones or lumbar pain-hematuria syndrome. Furthermore, because there are no systemic symptoms, the presence of hearing or visual impairment should prompt investigation for hereditary nephropathies, such as Alport syndrome. Some people with TBMD are thought to be carriers of the gene that causes Alport syndrome.

[0206] O. Mesangial proliferative glomerulonephritis Mesangial proliferative glomerulonephritis is a form of glomerulonephritis that primarily involves the mesangium. There is some evidence that interleukin-10 inhibits it in animal models. [2] The World Health Organization (WHO) classifies it as type II lupus nephritis. Mesangial cells in the renal glomerulus use endocytosis to internalize and degrade circulating immunoglobulins. This normal process promotes mesangial cell proliferation and matrix deposition. Therefore, when circulating immunoglobulin concentrations are elevated (i.e., in lupus and IgA nephropathy), the number of mesangial cells and matrix within the glomerulus is expected to increase, which is a hallmark of nephritic syndromes.

[0207] P. Amyloidosis (primary) Amyloidosis is a group of diseases in which abnormal proteins known as amyloid fibrils accumulate in tissues.[4] Symptoms vary depending on the type and often vary.[2] They may include diarrhea, weight loss, fatigue, tongue enlargement, bleeding, numbness, fainting when standing, swelling of the feet, or an enlarged spleen.[2] There are approximately 30 different types of amyloidosis, each resulting from the misfolding of a specific protein.[5] Some are hereditary, while others are acquired.[3] They are classified as localized and systemic forms.[2] The four most common types of systemic disease are light chain (AL), inflammatory (AA), dialysis (Aβ2M), hereditary, and elderly (ATTR) amyloidosis. Primary amyloidosis refers to amyloidosis without an associated clinical condition.

[0208] Q. c1q nephropathy C1q nephropathy is a rare glomerular disease characterized by characteristic mesangial C1q deposits on immunofluorescence microscopy. This nephropathy is histologically defined and poorly understood. Light microscopic features are heterogeneous and include minimal change nephrotic syndrome (MCD), focal segmental glomerulosclerosis (FSGS), and proliferative glomerulonephritis. Clinical presentations are also diverse, ranging from asymptomatic hematuria or proteinuria to frank nephritis or nephrotic syndrome in both children and adults. Elevated blood pressure and renal insufficiency at diagnosis are common findings. Optimal treatment is unclear and is usually guided by the underlying light microscopic lesion. Corticosteroids are the mainstay of treatment, with immunosuppressants reserved for steroid-resistant cases. The presence of nephrotic syndrome and FSGS appears to predict adverse outcomes in contrast to the favorable outcomes in MCD patients. (Devasahayam, et al., “C1q Nephropathy: The Unique Underrecognized Pathological Entity,” Analytical Cellular Pathology, vol. 2015, Article ID 490413, 5 pages, 2015. https: / / doi.org / 10.1155 / 2015 / 490413.) R. Anti-GBM disease Anti-glomerular basement membrane (GBM) disease, also known as Goodpasture's disease, is a rare condition that causes inflammation of small blood vessels in the kidneys and lungs. Although anti-glomerular basement membrane (GBM) antibodies primarily attack the kidneys and lungs, systemic symptoms such as fatigue, weight loss, tiredness, fever, and chills are also common, as well as joint aches and pains. 60–80% of patients with this condition experience both pulmonary and renal abnormalities, 20–40% have renal abnormalities alone, and less than 10% have pulmonary abnormalities alone. Pulmonary symptoms usually precede renal symptoms and typically include hemoptysis, chest pain (in less than 50% of cases), cough, and shortness of breath. Renal symptoms typically include hematuria, proteinuria, unexplained swelling of the limbs or face, excessive blood urea, and high blood pressure. GPS causes abnormal production of anti-GBM antibodies by blood plasma cells. Anti-GBM antibodies attack the alveoli and glomerular basement membrane. These antibodies bind reactive epitopes to the basement membrane, activating the complement cascade and killing targeted cells. T cells are also involved. The disease is generally considered a type II hypersensitivity reaction.

[0209] S. Polycystic kidney disease Polycystic kidney disease (PKD) is a rare, progressive kidney disease and a leading cause of chronic kidney disease. PKD accounts for 7% to 10% of end-stage renal disease (ESRD) cases. Approximately half of all PKD patients progress to ESRD by age 40 to 60. PKD affects people of all races, and men typically experience a slightly more aggressive disease. There are two major categories of PKD: autosomal dominant PKD (ADPKD) and autosomal recessive PKD (ARPKD). The former is more common, while the latter is a pediatric disease typically characterized by a more severe and accelerated disease course. Renal cysts are a defining feature of PKD. Patients with PKD are at increased risk for elevated blood pressure, CV events, aneurysms, liver cysts, pyelonephritis, and pain.

[0210] Measurement of urine protein levels Urinary protein levels can be measured using methods known in the art. Until recently, accurate protein measurements required a 24-hour urine collection. In a 24-hour collection, patients urinate into a container and refrigerate it after each toilet visit. Patients are instructed to begin collecting urine after their first toilet visit in the morning. All remaining urine for the day is collected in the container. The next morning, the patient completes the collection with the first urine they urinate upon waking.

[0211] More recently, researchers have found that a single urine sample can provide the necessary information. Newer methods compare the amount of albumin in a urine sample with the amount of creatinine, a waste product formed by the breakdown of normal muscle. This measurement is called the urinary albumin-to-creatinine ratio (UACR). A urine sample containing more than 30 milligrams of albumin per gram of creatinine (30 mg / g) signals a potential problem. If the lab value exceeds 30 mg / g, another UACR test should be performed in 1 to 2 weeks. If the second test also shows high levels of protein, the patient has persistent proteinuria, a sign of declining kidney function, and should undergo additional testing to evaluate kidney function.

[0212] Tests that measure the amount of creatinine in the blood also indicate whether a person's kidneys are efficiently removing waste products. Excess creatinine in the blood is a sign of kidney damage. Doctors can use creatinine measurements to estimate how efficiently the kidneys are filtering blood. This calculation is called the estimated glomerular filtration rate (or eGFR). Chronic kidney disease is present when the eGFR is less than 60 milliliters per minute (mL / min).

[0213] TRPC5 TRPCs are a family of animal transient receptor potential cation channels. TRPC5 is a subtype of the TRPC family of mammalian transient receptor potential ion channels. Three examples of TRPC5 are shown in Table 1 below.

[0214] [Table 5]

[0215] Accordingly, in certain embodiments, the present invention provides a method for treating or reducing the risk of developing a disease or condition selected from renal disease, pulmonary arterial hypertension, anxiety, depression, cancer, diabetic retinopathy, or pain, comprising administering to a subject in need thereof a therapeutically effective amount of a compound of the present invention (e.g., a compound of structural formula I) or a pharmaceutical composition comprising the compound.

[0216] In some embodiments, the disease is kidney disease, anxiety, depression, cancer, or diabetic retinopathy.

[0217] In some embodiments, the disease or condition is a renal disease selected from focal segmental glomerulosclerosis (FSGS), diabetic nephropathy, Alport syndrome, hypertensive renal disease, nephrotic syndrome, steroid-resistant nephrotic syndrome, minimal change nephrotic syndrome, membranous nephropathy, idiopathic membranous nephropathy, membranoproliferative glomerulonephritis (MPGN), immune complex-mediated MPGN, complement-mediated MPGN, lupus nephritis, post-infectious glomerulonephritis, thin basement membrane disease, mesangial proliferative glomerulonephritis, amyloidosis (primary), c1q nephritis, rapidly progressive GN, anti-GBM disease, C3 glomerulonephritis, hypertensive nephrosclerosis, or IgA nephropathy. In some embodiments, the renal disease is a proteinuric renal disease. In some embodiments, the renal disease is microalbuminuric or macroalbuminuric renal disease.

[0218] In some embodiments, the disease or condition being treated is pulmonary arterial hypertension.

[0219] In some embodiments, the disease or condition being treated is pain selected from neuropathic pain and visceral pain.

[0220] In some embodiments, the disease or condition is a cancer selected from chemotherapy-resistant breast cancer, adriamycin-resistant breast cancer, chemotherapy-resistant colorectal cancer, medulloblastoma, and tumor angiogenesis.

[0221] The present invention also provides a method for treating or reducing the risk of developing anxiety, or depression, or cancer, comprising administering to a subject in need thereof a therapeutically effective amount of a compound of the present invention (e.g., a compound of Formula I) or a pharmaceutical composition comprising said compound.

[0222] In some embodiments, the disease or condition being treated is transplant-associated FSGS, transplant-associated nephrotic syndrome, transplant-associated proteinuria, cholestatic liver disease, polycystic kidney disease, autosomal dominant polycystic kidney disease (ADPKD), obesity, insulin resistance, type II diabetes, prediabetes, metabolic syndrome, nonalcoholic fatty liver disease (NAFLD), or nonalcoholic steatohepatitis (NASH).

[0223] Treatment target In one embodiment of the present invention, the subject is selected based on having or being at risk of developing kidney disease, pulmonary arterial hypertension, anxiety, depression, cancer, diabetic retinopathy, or pain. In another embodiment, the subject is selected based on having or being at risk of developing kidney disease, anxiety, depression, cancer, or diabetic retinopathy. In another embodiment of the present invention, the subject is selected based on having or being at risk of developing pain, neuropathic pain, visceral pain, transplant-associated FSGS, transplant-associated nephrotic syndrome, transplant-associated proteinuria, cholestatic liver disease, polycystic kidney disease, autosomal dominant polycystic kidney disease (ADPKD), obesity, insulin resistance, type II diabetes, prediabetes, metabolic syndrome, nonalcoholic fatty liver disease (NAFLD), or nonalcoholic steatohepatitis (NASH).

[0224] Subjects with or at risk of developing proteinuria include those with diabetes, elevated blood pressure, or certain family backgrounds. Diabetes is the leading cause of end-stage renal disease (ESRD) in the United States. In both type 1 and type 2 diabetes, urinary albumin is one of the first signs of declining kidney function. As kidney function declines, the amount of albumin in urine increases. Another risk factor for developing proteinuria is elevated blood pressure. Proteinuria in hypertensive patients is an indicator of declining kidney function. Uncontrolled elevated blood pressure can lead to complete kidney failure. African Americans are more likely than Caucasians to develop high blood pressure and are more likely to experience kidney problems even when blood pressure is only mildly elevated. Other groups at risk for proteinuria include Native Americans, Hispanics / Latinos, Pacific Islanders, the elderly, and overweight subjects.

[0225] In one embodiment of the present invention, a subject is selected based on having or being at risk of developing proteinuria. A subject with or at risk of developing proteinuria is one who has one or more symptoms of the condition. Symptoms of proteinuria are known in the art and include, but are not limited to, large amounts of protein in the urine. Protein in the urine causes the urine to appear foamy in the toilet. Large amounts of protein loss can result in edema, which can cause swelling of the hands, feet, abdomen, or face. These are signs of large amounts of protein loss and indicate the progression of kidney disease. Laboratory tests are the only way to determine whether a subject has protein in their urine before extensive kidney damage occurs.

[0226] The methods are effective in a variety of subjects, including mammals, e.g., humans, and other animals, e.g., laboratory animals, e.g., mice, rats, rabbits, or monkeys, or pets and livestock, e.g., cats, dogs, goats, sheep, pigs, cows, or horses. In some embodiments, the subject is a mammal. In some embodiments, the subject is a human. [Example]

[0227] The present invention is further described in the following examples, which do not limit the scope of the invention as defined in the claims.

[0228] Example 1 Synthesis of Compound 100

[0229] [ka] tert-Butyl 4-[4-fluoro-2-(trifluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate To a stirred solution of tert-butyl 4-chloro-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (400 mg, 1.48 mmol, 1 equiv.) and 4-fluoro-2-(trifluoromethyl)phenol (400.6 mg, 2.22 mmol, 1.5 equiv.) in acetonitrile (10 mL) was added DBU (451.5 mg, 2.97 mmol, 2.00 equiv.) at room temperature. The resulting mixture was stirred at 80° C. for 2 hours. The reaction was monitored by LCMS. The mixture was allowed to cool to room temperature. The resulting mixture was concentrated under reduced pressure. The resulting mixture was extracted with DCM (3×100 mL). The combined organic layers were washed with brine (3×100 mL) and dried over anhydrous NaSO. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC (PE / EtOAc=2:1) ​​to give tert-butyl 4-[4-fluoro-2-(trifluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (110 mg, 17.94%) as a brown solid.

[0230] 4-[4-fluoro-2-(trifluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine To a stirred solution of tert-butyl 4-[4-fluoro-2-(trifluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (110 mg, 0.27 mmol, 1 equiv.) in DCM (4 mL) was added TFA (1 mL, 13.46 mmol, 50.59 equiv.) at room temperature. The resulting mixture was stirred at room temperature for 1 hour. The reaction was monitored by LCMS. The resulting mixture was concentrated under reduced pressure. The mixture was basified to pH 8 with saturated NaHCO3 (aq.). The resulting mixture was concentrated under reduced pressure. The residue was purified by preparative TLC (DCM / MeOH=12:1) to afford 4-[4-fluoro-2-(trifluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine (50 mg, 59.98%) as a brown solid.

[0231] 4-chloro-5-[4-[4-fluoro-2-(trifluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one To a stirred solution of 4-[4-fluoro-2-(trifluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine (50 mg, 0.16 mmol, 1 equiv.) in DIEA (2 mL) was added 4,5-dichloro-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (47.5 mg, 0.19 mmol, 1.19 equiv.) at room temperature. The resulting mixture was stirred at 100° C. for 2 hours. The reaction was monitored by LCMS. The mixture was allowed to cool to room temperature. The residue was purified by preparative TLC (PE / EtOAc=2:1) ​​to give 4-chloro-5-[4-[4-fluoro-2-(trifluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (40 mg, 47.65%) as a brown solid.

[0232] 4-chloro-5-[4-[4-fluoro-2-(trifluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2,3-dihydropyridazin-3-one To a stirred solution of 4-chloro-5-[4-[4-fluoro-2-(trifluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (40 mg, 0.08 mmol, 1 equiv.) in DCM (4 mL) was added TFA (1 mL, 13.46 mmol, 177.00 equiv.) dropwise at room temperature. The resulting mixture was stirred at room temperature for 1 hour. The reaction was monitored by LCMS. The resulting mixture was concentrated under reduced pressure. The mixture was basified to pH 8 with saturated NaHCO3 (aq.). The resulting mixture was concentrated under reduced pressure. The crude product (40 mg) was purified by preparative HPLC using the following conditions: (Column: XBridge Prep OBD C18 column, 30 × 150 mm, 5 μm, Mobile phase A = water (10 mmol / L NH4HCO3), Mobile phase B = acetonitrile, Flow rate = 60 mL / min, Gradient = 18% B to 47% B in 7 min, 220 nm, Rt = 6.22 min) to give 4-chloro-5-[4-[4-fluoro-2-(trifluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2,3-dihydropyridazin-3-one (8.6 mg, 25.59%) as a white solid.

[0233] Example 2 Synthesis of Compound 140

[0234] [ka] tert-Butyl 4-bromo-5,6,7,8-tetrahydro-1,7-naphthyridine-7-carboxylate To a solution of 4-bromo-5,6,7,8-tetrahydro-1,7-naphthyridine (250 mg, 1.173 mmol, 1 equiv) in THF (10 mL, 123.430 mmol, 105.20 equiv) was added BocO (512.13 mg, 2.347 mmol, 2.00 equiv) and TEA (474.90 mg, 4.693 mmol, 4 equiv) at 25 °C. The solution was stirred at 25 °C for 2 h. The resulting mixture was concentrated under reduced pressure. The residue was purified by preparative TLC (PE / EA = 5 / 1) to give tert-butyl 4-bromo-5,6,7,8-tetrahydro-1,7-naphthyridine-7-carboxylate (210 mg, 57.15%) as a pale yellow oil.

[0235] tert-Butyl 4-[4-fluoro-2-(trifluoromethyl)phenoxy]-5,6,7,8-tetrahydro-1,7-naphthyridine-7-carboxylate To a solution of tert-butyl 4-bromo-5,6,7,8-tetrahydro-1,7-naphthyridine-7-carboxylate (210 mg, 0.671 mmol, 1 equiv.) and 4-fluoro-2-(trifluoromethyl)phenol (241.52 mg, 1.341 mmol, 2 equiv.) in DMSO (10 mL) was added CsCO (873.86 mg, 2.682 mmol, 4 equiv.), 2-(dimethylamino)acetic acid (41.46 mg, 0.402 mmol, 0.6 equiv.), and CuI (76.62 mg, 0.402 mmol, 0.60 equiv.). After stirring at 120 °C under a nitrogen atmosphere for 4 h, the resulting mixture was concentrated under reduced pressure. The residue was purified by preparative TLC, eluting with PE / EA (5 / 1) to give tert-butyl 4-[4-fluoro-2-(trifluoromethyl)phenoxy]-5,6,7,8-tetrahydro-1,7-naphthyridine-7-carboxylate (100 mg, 36.17%) as a pale yellow solid.

[0236] 4-[4-fluoro-2-(trifluoromethyl)phenoxy]-5,6,7,8-tetrahydro-1,7-naphthyridine To a solution of tert-butyl 4-[4-fluoro-2-(trifluoromethyl)phenoxy]-5,6,7,8-tetrahydro-1,7-naphthyridine-7-carboxylate (150 mg, 0.364 mmol, 1 equiv) in DCM (10 mL, 157.300 mmol, 432.46 equiv) was added TFA (414.75 mg, 3.637 mmol, 10 equiv) at 25° C. The solution was stirred at 25° C. for 2 hours. The resulting mixture was concentrated under reduced pressure. The residue was used in the next step.

[0237] 4-chloro-5-[4-[4-fluoro-2-(trifluoromethyl)phenoxy]-5,6,7,8-tetrahydro-1,7-naphthyridin-7-yl]-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one A mixture of 4-[4-fluoro-2-(trifluoromethyl)phenoxy]-5,6,7,8-tetrahydro-1,7-naphthyridine (60 mg, 0.192 mmol, 1 equiv.) and 4,5-dichloro-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (47.86 mg, 0.192 mmol, 1.00 equiv.) in DIEA (49.67 mg, 0.384 mmol, 2 equiv.) was stirred at 100° C. for 2 h under a N atmosphere. The residue was purified by preparative TLC (PE / EA=1 / 1) to give 4-chloro-5-[4-[4-fluoro-2-(trifluoromethyl)phenoxy]-5,6,7,8-tetrahydro-1,7-naphthyridin-7-yl]-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (100 mg, 99.15%) as a pale yellow solid.

[0238] 4-chloro-5-[4-[4-fluoro-2-(trifluoromethyl)phenoxy]-5,6,7,8-tetrahydro-1,7-naphthyridin-7-yl]-2,3-dihydropyridazin-3-one To a solution of 4-chloro-5-[4-[4-fluoro-2-(trifluoromethyl)phenoxy]-5,6,7,8-tetrahydro-1,7-naphthyridin-7-yl]-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (100 mg, 0.191 mmol, 1 equiv) in DCM (10 mL, 157.300 mmol, 825.67 equiv) was added TFA (217.23 mg, 1.905 mmol, 10.00 equiv) at 25° C. The solution was stirred at 25° C. for 2 h. The crude product (150 mg) was purified by preparative HPLC using the following conditions: (Column: XBridge Shield RP18 OBD column, 30 × 150 mm, 5 μm, Mobile phase A = water (10 mM NH4HCO3), Mobile phase B = acetonitrile, Flow rate = 60 mL / min, Gradient = 20% B to 40% B in 7 min, 220 nm, Rt = 6.63 min) to give 4-chloro-5-[4-[4-fluoro-2-(trifluoromethyl)phenoxy]-5,6,7,8-tetrahydro-1,7-naphthyridin-7-yl]-2,3-dihydropyridazin-3-one (42.9 mg, 51.09%) as a white solid.

[0239] Example 3 Synthesis of Compound 120

[0240] [ka] 2-Benzyl-5-[4-fluoro-2-(trifluoromethyl)phenoxy]-1,2,3,4-tetrahydro-2,6-naphthyridine To a stirred mixture of 2-benzyl-5-bromo-1,2,3,4-tetrahydro-2,6-naphthyridine (250 mg, 0.825 mmol, 1 equiv.) and 2-(dimethylamino)acetic acid (170.05 mg, 1.649 mmol, 2.00 equiv.) in DMSO (5 mL), 4-fluoro-2-(trifluoromethyl)phenol (89.10 mg, 0.495 mmol, 0.6 equiv.) and CuI (94.22 mg, 0.495 mmol, 0.6 equiv.) were added at room temperature. CsCO (1074.59 mg, 3.298 mmol, 4 equiv.) was then added at room temperature. The final reaction mixture was irradiated at 120 °C for 1 h by microwave irradiation. The reaction was monitored by LCMS. The mixture was allowed to cool to room temperature. The crude product was purified by reverse-phase flash using the following conditions: (Column = XBridge Prep OBD C18 column, 30 x 150 mm, 5 um, Mobile phase A = water (10 mM NH4HCO3), Mobile phase B = acetonitrile, Flow rate = 60 mL / min, Gradient = 18% B to 35% B in 8 min, 220 nm, Rt = 7.12 min) to give 2-benzyl-5-[4-fluoro-2-(trifluoromethyl)phenoxy]-1,2,3,4-tetrahydro-2,6-naphthyridine (180 mg, 54.25%) as a brown solid.

[0241] 5-[4-Fluoro-2-(trifluoromethyl)phenoxy]-1,2,3,4-tetrahydro-2,6-naphthyridine To a stirred solution of 2-benzyl-5-[4-fluoro-2-(trifluoromethyl)phenoxy]-1,2,3,4-tetrahydro-2,6-naphthyridine (180 mg) in MeOH (10 mL) was added Pd / C (20 mg) at room temperature under a nitrogen atmosphere. The resulting mixture was stirred at room temperature under a hydrogen atmosphere for 5 hours. The reaction was monitored by LCMS. The resulting mixture was concentrated under reduced pressure. The residue was purified by preparative TLC (DCM / MeOH=12:1) to give 5-[4-fluoro-2-(trifluoromethyl)phenoxy]-1,2,3,4-tetrahydro-2,6-naphthyridine (100 mg) as a brown solid.

[0242] 4-chloro-5-[5-[4-fluoro-2-(trifluoromethyl)phenoxy]-1,2,3,4-tetrahydro-2,6-naphthyridin-2-yl]-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one To a stirred solution of 5-[4-fluoro-2-(trifluoromethyl)phenoxy]-1,2,3,4-tetrahydro-2,6-naphthyridine (100 mg, 0.320 mmol, 1 equiv.) in DIEA (0.1 mL) was added 4,5-dichloro-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (63.81 mg, 0.256 mmol, 0.8 equiv.) at room temperature. The resulting mixture was stirred at 90° C. for 1 hour. The reaction was monitored by LCMS. The mixture was allowed to cool to room temperature. The residue was purified by preparative TLC (DCM / MeOH=12:1) to give 4-chloro-5-[5-[4-fluoro-2-(trifluoromethyl)phenoxy]-1,2,3,4-tetrahydro-2,6-naphthyridin-2-yl]-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (130 mg, 77.34%) as a white solid.

[0243] 4-chloro-5-[5-[4-fluoro-2-(trifluoromethyl)phenoxy]-1,2,3,4-tetrahydro-2,6-naphthyridin-2-yl]-2,3-dihydropyridazin-3-one To a stirred solution of 4-chloro-5-[5-[4-fluoro-2-(trifluoromethyl)phenoxy]-1,2,3,4-tetrahydro-2,6-naphthyridin-2-yl]-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (107 mg, 0.204 mmol, 1 equiv.) in DCM (4 mL) was added TFA (1 mL) at room temperature. The resulting mixture was stirred at room temperature for 1 hour. The reaction was monitored by LCMS. The mixture was basified to pH 7 with saturated NaHCO3 (aq.). The resulting mixture was concentrated under reduced pressure. The crude product (50 mg) was purified by preparative HPLC using the following conditions: (Column = XBridge Prep OBD C18 column, 30 × 150 mm, 5 μm, Mobile phase A = water (10 mM NH4HCO3), Mobile phase B = acetonitrile, Flow rate = 60 mL / min, Gradient = 30% B to 50% B in 8 min, 220 nm, Rt = 7.55 min) to give 4-chloro-5-[5-[4-fluoro-2-(trifluoromethyl)phenoxy]-1,2,3,4-tetrahydro-2,6-naphthyridin-2-yl]-2,3-dihydropyridazin-3-one (60 mg, 66.78%) as a white solid.

[0244] Example 4 Synthesis of Compound 118

[0245] [ka] Ethyl 2-(benzylamino)propanoate To a stirred solution of benzaldehyde (8 g, 75.384 mmol, 1 equiv.) and TEA (7.63 g, 75.384 mmol, 1 equiv.) in DCE (100 mL, 1263.149 mmol, 16.76 equiv.) was added TEA (7.63 g, 75.384 mmol, 1 equiv.) and NaBH(OAc) (31.95 g, 150.767 mmol, 2 equiv.) in a nitrogen atmosphere at room temperature in small portions. The mixture was stirred at room temperature overnight. The desired product could be detected by LCMS. The resulting mixture was extracted with DCM (2 × 150 mL). The combined organic layers were washed with brine (1 × 90 mL) and dried over anhydrous NaSO. After filtration, the filtrate was concentrated under reduced pressure to give ethyl 2-(benzylamino)propanoate (12 g, 76.80%) as a colorless oil.

[0246] Methyl 4-[benzyl(1-ethoxy-1-oxopropan-2-yl)amino]butanoate To a stirred solution of ethyl 2-(benzylamino)propanoate (8 g, 38.596 mmol, 1 equiv.) and methyl 4-oxobutanoate (4.48 g, 38.596 mmol, 1.00 equiv.) in DCE (120 mL, 1515.779 mmol, 39.27 equiv.), TEA (3.91 g, 38.596 mmol, 1 equiv.) and NaBH(OAc) (16.36 g, 77.193 mmol, 2 equiv.) were added portionwise at room temperature under a nitrogen atmosphere. The mixture was stirred overnight at room temperature. The desired product could be detected by LCMS. The resulting mixture was extracted with DCM (2 × 150 mL). The combined organic layers were washed with brine (1 × 90 mL) and dried over anhydrous NaSO. After filtration, the filtrate was concentrated under reduced pressure to give methyl 4-[benzyl(1-ethoxy-1-oxopropan-2-yl)amino]butanoate (10 g, 84.29%) as a colorless oil.

[0247] Methyl 1-benzyl-2-methyl-3-oxopiperidine-4-carboxylate To a stirred solution of methyl 4-[benzyl(1-ethoxy-1-oxopropan-2-yl)amino]butanoate (8 g, 26.026 mmol, 1 equiv.) in toluene (100 mL) was added t-BuOK (5.00 g, 52.051 mmol, 2 equiv.) at room temperature. The mixture was stirred at 80° C. for 2 hours. The desired product was detected by LCMS. The resulting mixture was concentrated in vacuo. The residue was purified by silica gel column chromatography eluting with PE / EtOAc (5:1 to 2:1) to give methyl 1-benzyl-2-methyl-3-oxopiperidine-4-carboxylate (6.5 g, 95.57%) as a white solid.

[0248] 7-benzyl-8-methyl-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-4-ol To a stirred solution of methyl 1-benzyl-2-methyl-3-oxopiperidine-4-carboxylate (6 g, 22.960 mmol, 1 equiv.) in EtOH (80 mL, 1377.083 mmol, 59.98 equiv.), t-BuONa (4.41 g, 45.921 mmol, 2 equiv.) and methanimidamide hydrochloride (3.70 g, 45.921 mmol, 2.00 equiv.) were added portionwise at room temperature under a nitrogen atmosphere. The mixture was stirred at 80° C. for 2 hours. The desired product could be detected by LCMS. The resulting mixture was concentrated in vacuo. The residue was purified by silica gel column chromatography eluting with PE / EtOAc (3:1 to 2:1) to give 7-benzyl-8-methyl-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-4-ol (5 g, 85.29%) as a white solid.

[0249] tert-Butyl 4-hydroxy-8-methyl-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate To a solution of 7-benzyl-8-methyl-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-4-ol (5 g, 19.583 mmol, 1 equiv.) in EtOH (60 mL, 1032.812 mmol, 52.74 equiv.), BocO (8.55 g, 39.166 mmol, 2 equiv.), CHCOONa (1.81 g, 23.500 mmol, 1.2 equiv.), Pd(OH) / C (275.01 mg, 1.958 mmol, 0.1 equiv.) were added under a nitrogen atmosphere. The mixture was hydrogenated under a hydrogen atmosphere at room temperature for 2 hours, filtered through a pad of Celite, and concentrated under reduced pressure to give tert-butyl 4-hydroxy-8-methyl-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (4.5 g, 86.61%) as a white solid.

[0250] tert-Butyl 4-chloro-8-methyl-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate To a stirred solution of tert-butyl 4-hydroxy-8-methyl-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (4.5 g, 16.961 mmol, 1 equiv.) and PPh3 (6.67 g, 25.442 mmol, 1.5 equiv.) in DCE (60 mL, 0.606 mmol, 0.04 equiv.), CCl4 (5.22 g, 33.922 mmol, 2 equiv.) was added in small portions at room temperature under a nitrogen atmosphere. The mixture was stirred at 70 °C for 2 h. The desired product could be detected by LCMS. The resulting mixture was concentrated in vacuo. The residue was purified by silica gel column chromatography eluting with PE / EtOAc (7:1) to give tert-butyl 4-chloro-8-methyl-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (4 g, 83.11%) as a white solid.

[0251] tert-Butyl 4-(2-chloro-4-fluorophenoxy)-8-methyl-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate To a stirred solution of tert-butyl 4-chloro-8-methyl-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (4 g, 14.096 mmol, 1 equiv.) and 2-chloro-4-fluorophenol (2.07 g, 14.096 mmol, 1 equiv.) in DMF (50 mL) was added KCO (3.90 g, 28.193 mmol, 2 equiv.) in portions at room temperature under a nitrogen atmosphere. The mixture was stirred at 70 °C for 1 h. The desired product could be detected by LCMS. The resulting mixture was concentrated in vacuo. The residue was purified by silica gel column chromatography, eluting with PE / EtOAc (1:1) to give tert-butyl 4-(2-chloro-4-fluorophenoxy)-8-methyl-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (4 g, 72.05%) as a white solid.

[0252] 4-(2-chloro-4-fluorophenoxy)-8-methyl-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine To a stirred solution of tert-butyl 4-(2-chloro-4-fluorophenoxy)-8-methyl-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (4 g, 1 equiv.) in DCM (20 mL) was added TFA (4 mL) dropwise / portionwise at room temperature under a nitrogen atmosphere. The mixture was stirred at room temperature for 2 hours. The desired product could be detected by LCMS. The resulting mixture was concentrated under reduced pressure to give 4-(2-chloro-4-fluorophenoxy)-8-methyl-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine (2.7 g, 90.51%) as an off-white solid.

[0253] 4-chloro-5-[4-(2-chloro-4-fluorophenoxy)-8-methyl-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one To a stirred solution of 4-(2-chloro-4-fluorophenoxy)-8-methyl-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine (1 g, 3.404 mmol, 1 equiv.) in DIEA (1 mL) was added 4,5-dichloro-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (0.85 g, 3.404 mmol, 1 equiv.) in portions at room temperature under a nitrogen atmosphere. The mixture was stirred at 100° C. overnight. The desired product could be detected by LCMS. The residue was purified by silica gel column chromatography, eluting with PE / EtOAc (1:1 to 1:2) to give 4-chloro-5-[4-(2-chloro-4-fluorophenoxy)-8-methyl-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (1 g, 58.01%) as a white solid.

[0254] 4-chloro-5-[4-(2-chloro-4-fluorophenoxy)-8-methyl-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2,3-dihydropyridazin-3-one To a stirred solution of 4-chloro-5-[4-(2-chloro-4-fluorophenoxy)-8-methyl-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (1 g, 1 equivalent) in DCM (10 mL) was added dropwise TFA (2 mL) at room temperature under a nitrogen atmosphere. The mixture was stirred at room temperature for 1 hour. The desired product could be detected by LCMS. The resulting mixture was concentrated under reduced pressure to give 4-chloro-5-[4-(2-chloro-4-fluorophenoxy)-8-methyl-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2,3-dihydropyridazin-3-one (600 mg, 71.95%) as a white solid.

[0255] 4-chloro-5-[(8R)-4-(2-chloro-4-fluorophenoxy)-8-methyl-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2,3-dihydropyridazin-3-one 4-Chloro-5-[4-(2-chloro-4-fluorophenoxy)-8-methyl-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2,3-dihydropyridazin-3-one (250 mg, 1 equiv.) was purified by preparative chiral HPLC (column: CHIRALPAK Separation by IG, 20 x 250 mm, 5 µm, mobile phase A = Hex:DCM = 3:1 (0.1% FA) - HPLC, mobile phase B = EtOH - HPLC, flow rate = 20 mL / min, gradient = 15B to 15B in 19 min, 220 / 254 nm, RT1 = 13.016, RT2 = 16.004) gave 4-chloro-5-[(8R)-4-(2-chloro-4-fluorophenoxy)-8-methyl-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2,3-dihydropyridazin-3-one (144 mg, 57.60%) as a white solid.

[0256] Example 5 Synthesis of Compound 103

[0257] [ka] tert-Butyl 4-[2-(difluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate To a stirred solution of tert-butyl 4-chloro-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (800 mg, 2.966 mmol, 1 equiv.) and 2-(difluoromethyl)phenylacetic acid (1104.26 mg, 5.932 mmol, 2.00 equiv.) in DMF (20 mL) was added K2CO3 (1229.72 mg, 8.898 mmol, 3 equiv.) in portions at 80 °C under a nitrogen atmosphere. The mixture was stirred for 2 h. The reaction was monitored by LCMS. The reaction was quenched with water at room temperature. The mixture was extracted with EtOAc (3 × 50 mL). The combined organic layers were washed with brine (3 × 100 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography using the following conditions: column = C18 silica gel, mobile phase = aqueous MeOH, gradient 10% to 50% in 10 min, detector = UV 254 nm to give tert-butyl 4-[2-(difluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (900 mg, 80.41%) as an off-white solid.

[0258] 4-[2-(difluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine To a stirred solution of tert-butyl 4-[2-(difluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (900 mg, 2.385 mmol, 1 equiv.) in DCM, 3,3,3-trifluoropropanoic acid (3 mL, 6.00 equiv.) was added dropwise at room temperature. The mixture was stirred for 1.5 h. The reaction was monitored by TLC (PE / EtOAc = 10:1). The residue was basified to pH 8 with saturated NaHCO3 (aq.). The mixture was concentrated under reduced pressure. The crude product (100 mg) was purified by preparative HPLC using the following conditions to afford 4-[2-(difluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine (329 mg, 49.75%) as an off-white solid.

[0259] 4-chloro-5-[4-[2-(difluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one To a stirred solution of 4-[2-(difluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine (328 mg, 1.183 mmol, 1 equiv) and 4,5-dichloro-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (169.05 mg, 0.679 mmol, 1.00 equiv) was added DIEA (175.43 mg, 1.357 mmol, 2.00 equiv) in small portions at 70° C. The mixture was stirred at 70° C. for 2 h. The residue was purified by reverse-phase flash chromatography using the following conditions: column = C18 silica gel, mobile phase = aqueous MeOH, gradient 10% to 50% in 10 min, detector = UV 254 nm to give 4-chloro-5-[4-[2-(difluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (328 mg, 56.60%) as an off-white solid.

[0260] 4-chloro-5-[4-[2-(difluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2,3-dihydropyridazin-3-one To a stirred solution of 4-chloro-5-[4-[2-(difluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (328 mg, 0.670 mmol, 1 equiv.) in DCM (10 mL) was added trifluoroacetic acid (3 mL) dropwise at room temperature. The mixture was concentrated in vacuo. The product was purified by preparative HPLC to give 4-chloro-5-[4-[2-(difluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2,3-dihydropyridazin-3-one (256.4 mg, 94.38%) as an off-white solid.

[0261] Example 6 Synthesis of Compounds 117 and 117a

[0262] [ka] Ethyl 4-[(1-phenylethyl)amino]pentanoate To a stirred solution of 1-phenylethan-1-amine (25 g, 206.300 mmol, 1 equiv.) and ethyl 4-oxopentanoate (29.74 g, 206.300 mmol, 1 equiv.) in DCE (400 mL, 5052.598 mmol, 24.49 equiv.) was added NaBH(OAc) (65.59 g, 309.449 mmol, 1.5 equiv.) in portions at 25 °C under a nitrogen atmosphere. The solution was stirred at 25 °C for 2 h. The reaction was quenched by the addition of HO (400 mL) at 0 °C. The resulting mixture was extracted with DCM (3 × 200 mL). The combined organic layers were washed with saturated NaCl (aq.) (3 × 200 mL) and dried over anhydrous NaSO. After filtration, the filtrate was concentrated under reduced pressure. The crude product was used in the next step.

[0263] Ethyl 4-[(2-ethoxy-2-oxoethyl)(1-phenylethyl)amino]pentanoate To a stirred solution of ethyl 4-[(1-phenylethyl)]pentanoate (49 g, 196.508 mmol, 1 equiv.) and ethyl 2-oxoacetate (40.12 g, 392.990 mmol, 2.00 equiv.) in DCE (500 mL, 6315.747 mmol, 32.14 equiv.) was added NaBH(OAc) (62.47 g, 294.762 mmol, 1.5 equiv.) in portions at 25 °C under a nitrogen atmosphere. The solution was stirred at 25 °C for 2 h. The reaction was quenched by the addition of HO (400 mL) at 0 °C. The resulting mixture was extracted with DCM (3 × 200 mL). The combined organic layers were washed with saturated NaCl (aq.) (3 × 200 mL) and dried over anhydrous NaSO. After filtration, the filtrate was concentrated under reduced pressure. The crude product, ethyl 4-[(2-ethoxy-2-oxoethyl)(1-phenylethyl)amino]pentanoate (57 g, 86.47%), was used in the next step.

[0264] Ethyl 2-methyl-5-oxo-1-(1-phenylethyl)piperidine-4-carboxylate To a solution of ethyl 4-[(2-ethoxy-2-oxoethyl)(1-phenylethyl)amino]pentanoate (57 g, 169.924 mmol, 1 equiv.) in toluene (500 mL, 4699.452 mmol, 27.66 equiv.), t-BuOK (47.67 g, 424.810 mmol, 2.5 equiv.) was added via a port at 0° C. The mixture was stirred at 25° C. for 2 hours. The resulting mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with PE / EA (50 / 1 to 10 / 1) to give ethyl 2-methyl-5-oxo-1-(1-phenylethyl)piperidine-4-carboxylate (29 g, 58.98%) as a yellow oil.

[0265] 7-(1-Cyclohexylethyl)-6-methyl-decahydropyrido[3,4-d]pyrimidin-4-ol To a solution of ethyl 2-methyl-5-oxo-1-(1-phenylethyl)piperidine-4-carboxylate (10 g, 34.557 mmol, 1 equiv.) and methanimidamide hydrochloride (4.17 g, 51.836 mmol, 1.50 equiv.) in EtOH (100 mL, 1721.353 mmol, 49.81 equiv.), EtONa (5.88 g, 86.393 mmol, 2.50 equiv.) was added via a port at 25 °C. The mixture was stirred at 90 °C for 2 h. The residue was purified by silica gel column chromatography eluting with DCM / MeOH (20 / 1 to 10 / 1) to give 7-(1-cyclohexylethyl)-6-methyl-decahydropyrido[3,4-d]pyrimidin-4-ol (3.4 g, 34.96%) as a yellow solid.

[0266] tert-Butyl 4-hydroxy-6-methyl-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate To a solution of 6-methyl-7-(1-phenylethyl)-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-4-ol (3.5 g, 12.994 mmol, 1 equiv.), HCOONH (4.10 g, 65.022 mmol, 5.00 equiv.), and BocO (8.51 g, 38.983 mmol, 3 equiv.) in EtOH (50 mL, 860.677 mmol, 66.23 equiv.) was added Pd(OH) / C (0.36 g, 2.599 mmol, 0.2 equiv.) under a nitrogen atmosphere. The mixture was hydrogenated under a hydrogen atmosphere using a hydrogen balloon at 70 °C for 2 h, filtered through a Celite pad, and concentrated under reduced pressure. Obtained tert-butyl 4-hydroxy-6-methyl-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (1.8 g, 52.21%) as a yellow solid.

[0267] tert-Butyl 4-chloro-6-methyl-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate To a solution of tert-butyl 4-hydroxy-6-methyl-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (1.8 g, 6.784 mmol, 1 equiv.) and PPh (3.56 g, 13.569 mmol, 2 equiv.) in DCE (20 mL, 252.630 mmol, 37.24 equiv.) was added CCl (3.13 g, 20.353 mmol, 3 equiv.) at 25 °C. The mixture was stirred at 70 °C for 3 h. The resulting mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with PE / EA (10 / 1 to 1 / 1) to give tert-butyl 4-chloro-6-methyl-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (1.1 g, 57.14%) as a yellow solid.

[0268] tert-Butyl 4-(2-chloro-4-fluorophenoxy)-6-methyl-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate To a solution of tert-butyl 4-chloro-6-methyl-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (1.1 g, 3.877 mmol, 1 equiv.) and 2-chloro-4-fluorophenol (0.85 g, 5.800 mmol, 1.50 equiv.) in DMF (15 mL, 193.826 mmol, 50.00 equiv.) was added KCO (1.07 g, 7.753 mmol, 2 equiv.) at 25 °C. The mixture was stirred at 70 °C for 1 h. The resulting mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with PE / EA (10 / 1 to 5 / 1) to give tert-butyl 4-(2-chloro-4-fluorophenoxy)-6-methyl-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (1.2 g, 78.60%) as a yellow solid.

[0269] 4-chloro-5-[4-(2-chloro-4-fluorophenoxy)-6-methyl-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one A mixture of 4-(2-chloro-4-fluorophenoxy)-6-methyl-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine (800 mg, 2.724 mmol, 1 equiv.) and 4,5-dichloro-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (678.42 mg, 2.724 mmol, 1.00 equiv.) in DIEA (704.01 mg, 5.447 mmol, 2 equiv.) was stirred at 100° C. for 16 h under a nitrogen atmosphere. The residue was purified by preparative TLC (PE / EA=1 / 1) to give 4-chloro-5-[4-(2-chloro-4-fluorophenoxy)-6-methyl-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (530 mg, 38.43%) as a pale yellow solid.

[0270] 4-chloro-5-[(6R)-4-(2-chloro-4-fluorophenoxy)-6-methyl-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2,3-dihydropyridazin-3-one To a solution of 4-chloro-5-[4-(2-chloro-4-fluorophenoxy)-6-methyl-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (530 mg, 1.047 mmol, 1 equiv) in DCM (20 mL, 314.601 mmol, 300.57 equiv), TFA (1193.47 mg, 10.467 mmol, 10 equiv) was added at 25° C. The solution was stirred at 25° C. for 2 hours. The resulting mixture was concentrated under reduced pressure. The crude product (600 mg) was purified by preparative HPLC using the following conditions: (Column: XBridge Shield RP18 OBD column, 30 × 150 mm, 5 μm, Mobile phase A: water (10 mM NH4HCO3), Mobile phase B: acetonitrile, Flow rate: 60 mL / min, Gradient: 20% B to 40% B in 7 min, 220 nm, Rt = 6.63 min) to give the racemate (200 mg). The residue (200 mg) was purified by chiral preparative HPLC using the following conditions: (Column: CHIRALPAK IE, 2 × 25 cm, 5 μm, Mobile phase A: MTBE (0.1% FA)-HPLC, Mobile phase B: IPA-HPLC, Flow rate: 18 mL / min, Gradient: 20% B to 20% B in 15 min, 220 / 254 nm). The two isomers were separated by this technique, but the absolute orientation was not determined. The compound designated 4-chloro-5-[(6S)-4-(2-chloro-4-fluorophenoxy)-6-methyl-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2,3-dihydropyridazin-3-one (60.9 mg, 13.78%) was obtained as a white solid in 9.688 minutes. The compound designated 4-chloro-5-[(6R)-4-(2-chloro-4-fluorophenoxy)-6-methyl-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2,3-dihydropyridazin-3-one (61.5 mg, 13.92%) was obtained as a white solid in 11.813 minutes.

[0271] Example 7 Synthesis of Compound 134

[0272] [ka] tert-Butyl 2-chloro-4-[2-(difluoromethyl)-4-fluorophenoxy]-5H,6H,7H,8H-pyrimido[3,4-d]pyrimidine-7-carboxylate To a stirred solution of 2-(difluoromethyl)-4-fluorophenol (5.33 g, 32.879 mmol, 2.00 equiv.) and tert-butyl 2,4-dichloro-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (5 g, 16.438 mmol, 1 equiv.) in DMF (30 mL) was added NaHCO (4.14 g, 49.282 mmol, 3.00 equiv.) at room temperature. The solution was stirred at 70 °C for 0.5 h. The mixture was concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography using the following conditions: column = spherical C18, 20-40 μm, 330 g, mobile phase A = water (+ 10 mM NH4HCO3), mobile phase B = acetonitrile, flow rate = 80 mL / min, gradient = 5% to 5% B in 10 min, gradient = 70% B to 95% B in 100 min, detector = 254 nm. Fractions containing the desired product were collected at 92% B and concentrated under reduced pressure to give tert-butyl 2-chloro-4-[2-(difluoromethyl)-4-fluorophenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (2.100 g) as an off-white solid.

[0273] 7-tert-butyl 2-methyl 4-[2-(difluoromethyl)-4-fluorophenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-2,7-dicarboxylate To a solution of tert-butyl 2-chloro-4-[2-(difluoromethyl)-4-fluorophenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (400 mg, 0.931 mmol, 1 equiv.) and TEA (188.34 mg, 1.861 mmol, 2 equiv.) in MeOH (15 mL, 370.484 mmol, 398.10 equiv.) was added Pd(PPh3)4 (107.54 mg, 0.093 mmol, 0.1 equiv.) in a pressure bath. The mixture was purged with nitrogen for 1 hour and then pressurized to 10 atmospheres with carbon monoxide at 100 °C for 16 hours. The reaction mixture was cooled to room temperature and filtered to remove insoluble solids. The residue was purified under the following conditions: column = spherical C 18 Purification was performed by reverse-phase flash chromatography using a 20-40 μm, 330 g, mobile phase A = water (+ 10 mM NHHCO), mobile phase B = acetonitrile, flow rate = 80 mL / min, gradient = 5% to 5% B in 10 min, 35% to 65% B in 20 min, detector = 254 nm. Fractions containing the desired product were collected at 62% B and concentrated under reduced pressure to give 7-tert-butyl 2-methyl 4-[2-(difluoromethyl)-4-fluorophenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-2,7-dicarboxylate (100 mg, 23.70%) as a colorless oil.

[0274] tert-Butyl 4-[2-(difluoromethyl)-4-fluorophenoxy]-2-(hydroxymethyl)-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate To a stirred solution of 7-tert-butyl 2-methyl 4-[2-(difluoromethyl)-4-fluorophenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-2,7-dicarboxylate (100 mg, 0.221 mmol, 1 equiv.) in t-BuOH (6 mL, 63.139 mmol, 286.29 equiv.), NaBH4 (16.69 mg, 0.441 mmol, 2 equiv.) was added at room temperature. The solution was stirred at 70 °C for 3 h. Water (3 mL) was added to the mixture. The residue was purified by reverse-phase flash chromatography using the following conditions: column = spherical C18, 20-40 μm, 330 g, mobile phase A = water (+ 10 mM NH4HCO3), mobile phase B = acetonitrile, flow rate = 80 mL / min, gradient = 5% to 5% B in 10 min, gradient = 45% B to 80% B in 20 min, detector = 254 nm. Fractions containing the desired product were collected at 74% B and concentrated under reduced pressure to give tert-butyl 4-[2-(difluoromethyl)-4-fluorophenoxy]-2-(hydroxymethyl)-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (35 mg, 37.30%) as a colorless oil.

[0275] [4-[2-(difluoromethyl)-4-fluorophenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-2-yl]methanol To a stirred solution of tert-butyl 4-[2-(difluoromethyl)-4-fluorophenoxy]-2-(hydroxymethyl)-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (35 mg) in DCM (6 mg) was added TFA (1 mg) at room temperature. The solution was stirred at room temperature for 2 hours. The mixture was concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography using the following conditions: column = spherical C18, 20-40 μm, 330 g, mobile phase A = water (+10 mM NH4HCO3), mobile phase B = acetonitrile, flow rate = 80 mL / min, gradient = 5% B to 5% B in 10 min, gradient = 25% B to 55% B in 20 min, detector = 254 nm. Fractions containing the desired product were collected at 41% B and concentrated under reduced pressure to give [4-[2-(difluoromethyl)-4-fluorophenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-2-yl]methanol (20 mg) as a colorless oil.

[0276] 4-chloro-5-[4-[2-(difluoromethyl)-4-fluorophenoxy]-2-(hydroxymethyl)-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one To a 25 mL round-bottom flask were added [4-[2-(difluoromethyl)-4-fluorophenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-2-yl]methanol (20 mg, 0.061 mmol, 1 equiv.) and 4,5-dichloro-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (15.31 mg, 0.061 mmol, 1 equiv.) at room temperature. To the mixture was added DIEA (15.89 mg, 0.123 mmol, 2 equiv.) at room temperature. The mixture was stirred at 90° C. for 2 hours. The residue was purified by reverse-phase flash chromatography using the following conditions: column = spherical C18, 20-40 μm, 330 g, mobile phase A = water (+ 10 mM NH4HCO3), mobile phase B = acetonitrile, flow rate = 80 mL / min, gradient = 5% to 5% B in 10 min, gradient = 35% B to 70% B in 20 min, detector = 254 nm. Fractions containing the desired product were collected at 65% B and concentrated under reduced pressure to give 4-chloro-5-[4-[2-(difluoromethyl)-4-fluorophenoxy]-2-(hydroxymethyl)-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (30 mg, 90.71%) as a colorless oil.

[0277] 4-chloro-5-[4-[2-(difluoromethyl)-4-fluorophenoxy]-2-(hydroxymethyl)-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2,3-dihydropyridazin-3-one To a solution of 4-chloro-5-[4-[2-(difluoromethyl)-4-fluorophenoxy]-2-(hydroxymethyl)-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (30 mg) in DCM (5 mL) was added TFA (1 mL) at room temperature. The solution was stirred at room temperature for 2 hours. The mixture was concentrated under reduced pressure. The crude product (30 mg) was purified by preparative HPLC using the following conditions: (Column: XBridge Prep OBD C18 column, 30 × 150 mm, 5 μm, Mobile phase A = unspecified, Mobile phase B = unspecified, Flow rate = 60 mL / min, Gradient = 20% B to 40% B in 8 min, 220 nm, Rt = 7.22 min) to give 4-chloro-5-[4-[2-(difluoromethyl)-4-fluorophenoxy]-2-(hydroxymethyl)-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2,3-dihydropyridazin-3-one (8.7 mg) as a white solid.

[0278] Compounds 128, 125, and 114 were prepared by the methods and schemes described in this example by substituting 2-trifluoromethylphenol, 4-fluoro-2-trifluoromethylphenol, and 4-fluoro-2-chlorophenol, respectively, for 2-(difluoromethyl)-4-fluorophenol in the first step of the synthesis.

[0279] Example 8 Synthesis of Compound 112

[0280] [ka] tert-Butyl 2-chloro-4-(2-chloro-4-fluorophenoxy)-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate To a stirred mixture of tert-butyl 2,4-dichloro-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (800 mg, 2.630 mmol, 1 equiv.) and 2-chloro-4-fluorophenol (578.16 mg, 3.945 mmol, 1.50 equiv.) in DMF (15 mL) was added K2CO3 (726.99 mg, 5.260 mmol, 2.00 equiv.) in portions at room temperature under a nitrogen atmosphere. The resulting mixture was stirred at 70 °C under a nitrogen atmosphere for 0.5 h. The reaction was monitored by LCMS. The mixture was allowed to cool to room temperature. The resulting mixture was concentrated under reduced pressure. The resulting mixture was extracted with EtOAc (3 × 200 mL). The combined organic layers were washed with brine (2 × 100 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with PE / EtOAc (30 / 1 to 10 / 1) to give tert-butyl 2-chloro-4-(2-chloro-4-fluorophenoxy)-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (1 g, 91.78%) as a yellow oil.

[0281] tert-Butyl 4-(2-chloro-4-fluorophenoxy)-2-[[(4-methoxyphenyl)methyl]amino]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate To a stirred mixture of tert-butyl 2-chloro-4-(2-chloro-4-fluorophenoxy)-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (700 mg, 1.690 mmol, 1 equiv.) in THF (30 mL) was added 1-(4-methoxyphenyl)methanamine (1159.02 mg, 8.449 mmol, 5.00 equiv.) in portions at room temperature under a nitrogen atmosphere. The resulting mixture was stirred at 60° C. for 16 hours under a nitrogen atmosphere. The reaction was monitored by LCMS. The mixture was allowed to cool to room temperature. The resulting mixture was concentrated under reduced pressure. The resulting mixture was extracted with EtOAc (3×200 mL). The combined organic layers were washed with brine (2×100 mL) and dried over anhydrous NaSO. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography using the following conditions: column: C18 silica gel, mobile phase: acetonitrile in water, gradient from 60% to 95% in 20 min, detector: UV 220 nm) to give tert-butyl 4-(2-chloro-4-fluorophenoxy)-2-[[(4-methoxyphenyl)methyl]amino]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (350 mg, 40.22%) as a yellow oil.

[0282] 4-(2-chloro-4-fluorophenoxy)-N-[(4-methoxyphenyl)methyl]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-2-amine To a stirred solution of tert-butyl 4-(2-chloro-4-fluorophenoxy)-2-[[(4-methoxyphenyl)methyl]amino]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (350 mg, 1 equiv.) in DCM (10 mL) was added TFA (1 mL) dropwise at room temperature. The reaction mixture was stirred at room temperature for 2 hours. The reaction was monitored by LCMS. The resulting mixture was concentrated under reduced pressure. The residue was basified to pH = 8 with saturated NH4HCO3 (aq.). The resulting mixture was extracted with DCM (3 x 100 mL). The combined organic layers were washed with brine (1 x 100 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative HPLC using the following conditions: (Column = XBridge Shield RP18 OBD column, 5 μm, 19 × 150 mm, Mobile phase A = water (10 mM NH4HCO3), Mobile phase B = acetonitrile, Flow rate = 25 mL / min, Gradient = 2% B to 32% B in 1 min, 220 / 254 nm, Rt = 7.08 min) to give 4-(2-chloro-4-fluorophenoxy)-N-[(4-methoxyphenyl)methyl]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-2-amine (260 mg) as a yellow oil.

[0283] 4-chloro-5-[4-(2-chloro-4-fluorophenoxy)-2-[[(4-methoxyphenyl)methyl]amino]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one A 50 mL round-bottom flask was charged with 4-(2-chloro-4-fluorophenoxy)-N-[(4-methoxyphenyl)methyl]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-2-amine (260 mg, 0.627 mmol, 1 equiv.), 4,5-dichloro-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (156.11 mg, 0.627 mmol, 1.00 equiv.), and DIEA (242.99 mg, 1.880 mmol, 3.00 equiv.) under a nitrogen atmosphere at room temperature. The resulting mixture was stirred at 90° C. for 2 hours under a nitrogen atmosphere. The residue was purified by reverse-phase flash chromatography using the following conditions: (column = C18 silica gel, mobile phase = aqueous acetonitrile, 50% to 85% gradient in 25 minutes, detector = UV 220 nm) to give 4-chloro-5-[4-(2-chloro-4-fluorophenoxy)-2-[[(4-methoxyphenyl)methyl]amino]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (350 mg, 89.00%) as a yellow solid.

[0284] 5-[2-amino-4-(2-chloro-4-fluorophenoxy)-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-4-chloro-2,3-dihydropyridazin-3-one To a stirred solution of 4-chloro-5-[4-(2-chloro-4-fluorophenoxy)-2-[[(4-methoxyphenyl)methyl]amino]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (200 mg) in TFA (8 mL, 107.704 mmol, 328.23 equiv.) was added 4-chloro-5-[4-(2-chloro-4-fluorophenoxy)-2-[[(4-methoxyphenyl)methyl]amino]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (200 mg). The final reaction mixture was irradiated by microwave irradiation at 80 °C for 2 h. The reaction was monitored by LCMS. The resulting mixture was concentrated under reduced pressure. The residue was basified to pH = 8 with saturated NH4HCO3 (aq). The resulting mixture was extracted with DCM (2 × 100 mL). The combined organic layers were washed with brine (1 × 100 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative HPLC using the following conditions: (Column = XBridge Prep OBD C18 column, 30 x 150 mm, 5 um, Mobile phase A = water (10 mM NH4HCO3), Mobile phase B = acetonitrile, Flow rate = 60 mL / min, Gradient = 25% B to 40% B in 8 min, 220 nm, Rt = 7.35 min) to give 5-[2-amino-4-(2-chloro-4-fluorophenoxy)-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-4-chloro-2,3-dihydropyridazin-3-one (52.4 mg) as a yellow solid.

[0285] Compounds 113, 116, and 102 were prepared by the methods and schemes described in this example by substituting 2-chlorophenol, 4-fluoro-2-trifluoromethylphenol, and 2-trifluorophenol, respectively, for 2-chloro-4-fluorophenol in the first step of the synthesis.

[0286] Example 9 Synthesis of Compounds 129 and 130

[0287] [ka] 1-[4-[4-fluoro-2-(trifluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-2-yl]ethan-1-one To a mixture of tert-butyl 2-chloro-4-[4-fluoro-2-(trifluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (600 mg, 1.340 mmol, 1 equiv.) and tributyl(1-ethoxyethenyl)stannane (967.80 mg, 2.680 mmol, 2.00 equiv.) in toluene (10 mL), Pd(PPh3)4 (77.41 mg, 0.067 mmol, 0.05 equiv.) was added at room temperature under a nitrogen atmosphere. The resulting mixture was stirred at 110 °C for 4 h. The reaction was monitored by LCMS. Tert-butyl 2-(1-ethoxyethenyl)-4-[4-fluoro-2-(trifluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (700 mg, 108.06%) was obtained as a yellow oil. The crude mixture obtained was used directly in the next step without further purification.

[0288] To a stirred solution of tert-butyl 2-(1-ethoxyethenyl)-4-[4-fluoro-2-(trifluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (1 g, 2.068 mmol, 1 equiv.) in DCM (5 mL) was added TFA (3.33 mL, 29.239 mmol, 21.70 equiv.) at room temperature. The resulting mixture was stirred at room temperature for 2 hours. The reaction was monitored by LCMS. The resulting mixture was concentrated under reduced pressure. The mixture / residue was basified to pH 8 with saturated NaHCO3 (aq.). The resulting mixture was concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography using the following conditions: column = spherical C18, 20-40 μm, 330 g, mobile phase A = water (+ 5 mM NH4HCO3), mobile phase B = acetonitrile, flow rate = 80 mL / min, gradient = 5% to 5% B in 10 min, gradient = 43% B to 55% B in 20 min, detector = 220 nm. Fractions containing the desired product were collected at 50% B and concentrated under reduced pressure to give 1-[4-[4-fluoro-2-(trifluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-2-yl]ethan-1-one (750 mg, 102.06%) as a pale yellow solid.

[0289] 5-[2-acetyl-4-[4-fluoro-2-(trifluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-4-chloro-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one To a 50 mL round-bottom flask, 1-[4-[4-fluoro-2-(trifluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-2-yl]ethan-1-one (750 mg, 2.111 mmol, 1 eq.) and 4,5-dichloro-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (525.81 mg, 2.111 mmol, 1.00 eq.) were added at room temperature. DIEA (818.47 mg, 6.333 mmol, 3.00 eq.) was added to the above mixture. The resulting mixture was stirred at 100° C. for 2 hours. The reaction was monitored by LCMS. The mixture was allowed to cool to room temperature. The residue was purified by reverse-phase flash chromatography using the following conditions: (Column: C18 spherical, 20-40 μm, 330 g; Mobile phase A: water (+ 5 mM NH4HCO3); Mobile phase B: acetonitrile; Flow rate: 80 mL / min; Gradient: 5% to 5% B in 10 min, 60% to 85% B in 20 min; Detector: 220 nm). Fractions containing the desired product were collected at 80% B and concentrated under reduced pressure to give 5-[2-acetyl-4-[4-fluoro-2-(trifluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-4-chloro-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (230 mg, 19.18%) as a pale yellow oil.

[0290] 4-chloro-5-[4-[4-fluoro-2-(trifluoromethyl)phenoxy]-2-(1-hydroxyethyl)-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one To a stirred solution of 5-[2-acetyl-4-[4-fluoro-2-(trifluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-4-chloro-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (230 mg, 0.405 mmol, 1 equiv.) in MeOH (10 mL) was added NaBH4 (30.64 mg, 0.810 mmol, 2.00 equiv.) in portions at 0° C. under a nitrogen atmosphere. The resulting mixture was stirred at room temperature for 2 hours. The reaction was monitored by LCMS. The resulting mixture was concentrated under reduced pressure. The residue was purified by preparative TLC (PE / EtOAc=1 / 1) to give 4-chloro-5-[4-[4-fluoro-2-(trifluoromethyl)phenoxy]-2-(1-hydroxyethyl)-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (120 mg, 51.99%) as a pale yellow oil.

[0291] 4-chloro-5-[4-[4-fluoro-2-(trifluoromethyl)phenoxy]-2-[(1S)-1-hydroxyethyl]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2,3-dihydropyridazin-3-one and 4-chloro-5-[4-[4-fluoro-2-(trifluoromethyl)phenoxy]-2-[(1R)-1-hydroxyethyl]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2,3-dihydropyridazin-3-one To a stirred solution of 4-chloro-5-[4-[4-fluoro-2-(trifluoromethyl)phenoxy]-2-(1-hydroxyethyl)-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (120 mg, 0.211 mmol, 1 equiv.) in DCM (5 mL) was added TFA (2.00 mL, 17.541 mmol, 127.89 equiv.) dropwise at room temperature. The resulting mixture was stirred at room temperature for 2 hours. The reaction was monitored by LCMS. The residue was basified to pH 8 with saturated NaHCO3 (aq.). The resulting mixture was concentrated under reduced pressure. The residue was purified using the following conditions: column = spherical C 18 The crude product (50 mg) was purified by reverse-phase flash chromatography using a 20-40 μm column, 330 g sample, mobile phase A = water (+ 5 mM NH4HCO3), mobile phase B = acetonitrile, flow rate = 80 mL / min, gradient = 5% to 5% B in 10 min, 40% B to 80% B in 25 min, detector = 220 nm). Fractions containing the desired product were collected at 55% B and concentrated under reduced pressure. The crude product (50 mg) was purified by chiral preparative HPLC using the following conditions: (Column = CHIRALPAK IE, 2 x 25 cm, 5 μm, mobile phase A = hexane (0.1% FA)-HPLC, mobile phase B = EtOH-HPLC, flow rate = 16 mL / min, gradient = 30% B to 30% B in 33 min, 220 / 254 nm, RT1 = 26.219, RT2 = 29.589). Two isomers were separated by this technique, but the absolute orientation was not determined. The compound designated 4-chloro-5-[4-[4-fluoro-2-(trifluoromethyl)phenoxy]-2-[(1S)-1-hydroxyethyl]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2,3-dihydropyridazin-3-one (27.1 mg) was obtained as an off-white solid in 29.589 minutes. The compound designated 4-chloro-5-[4-[4-fluoro-2-(trifluoromethyl)phenoxy]-2-[(1R)-1-hydroxyethyl]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2,3-dihydropyridazin-3-one (22.6 mg) was obtained as an off-white solid in 26.219 minutes.

[0292] Compound 119 was prepared by the methods and schemes described in this example, using tert-butyl 2-chloro-4-[4-fluoro-2-chlorophenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate as the starting material.

[0293] Compounds 122 and 123 were prepared by the methods and schemes described in this Example, starting with tert-butyl 2-chloro-4-[2-(trifluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate. Again, the absolute orientation of these separated isomers has not been determined, and the designation as (S) or (R) is arbitrary.

[0294] Example 10 Synthesis of Compound 115

[0295] [ka] tert-Butyl 2-chloro-4-[2-(trifluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate To a stirred solution of tert-butyl 2,4-dichloro-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (2 g, 6.58 mmol, 1 equiv.) and 2-(trifluoromethyl)phenol (1.6 g, 9.86 mmol, 1.5 equiv.) in acetonitrile (20 mL) was added DBU (2.0 g, 13.15 mmol, 2 equiv.) at room temperature. The solution was stirred at room temperature for 4 hours. The mixture was concentrated under reduced pressure. The residue was purified by preparative TLC (PE / EtOAc = 10:1) to afford tert-butyl 2-chloro-4-[2-(trifluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (700 mg, 24.77%) as a colorless oil.

[0296] tert-Butyl 2-([2-[(tert-butyldimethylsilyl)oxy]ethyl]amino)-4-[2-(trifluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate To a solution of tert-butyl 2-chloro-4-[2-(trifluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (500 mg, 1.163 mmol, 1 equiv.) in THF (15 mL), (2-aminoethoxy)(tert-butyl)dimethylsilane (1019.89 mg, 5.816 mmol, 5.00 equiv.) was added at room temperature under a nitrogen atmosphere. The resulting mixture was stirred at 50° C. for 16 hours. The reaction was monitored by LCMS. The resulting mixture was concentrated under reduced pressure. The residue was purified by preparative TLC (PE / EtOAc=3 / 1) to give tert-butyl 2-([2-[(tert-butyldimethylsilyl)oxy]ethyl]amino)-4-[2-(trifluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (440 mg, 66.51%) as a pale yellow oil.

[0297] 2-([4-[2-(trifluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-2-yl]amino)ethan-1-ol To a stirred solution of tert-butyl 2-([2-[(tert-butyldimethylsilyl)oxy]ethyl]amino)-4-[2-(trifluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (440 mg, 0.774 mmol, 1 equiv.) in DCM (10 mL) was added TFA (3 mL, 40.389 mmol, 52.20 equiv.) at room temperature. The resulting mixture was stirred at room temperature for 2 hours. The reaction was monitored by LCMS. The resulting mixture was concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography using the following conditions: column = C18 silica gel, mobile phase = aqueous ACN, 40% to 60% gradient in 15 min, detector = UV 254 nm) to give 2-([4-[2-(trifluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-2-yl]amino)ethan-1-ol (220 mg) as a pale yellow oil.

[0298] 4-chloro-5-[2-[(2-hydroxyethyl)amino]-4-[2-(trifluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one To a 50 mL round-bottom flask were added 2-([4-[2-(trifluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-2-yl]amino)ethan-1-ol (220 mg, 0.621 mmol, 1 eq.) and 4,5-dichloro-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (154.66 mg, 0.621 mmol, 1.00 eq.) at room temperature. To the above mixture was added DIEA (240.74 mg, 1.863 mmol, 3.00 eq.). The resulting mixture was stirred at 100° C. for 2 hours. The reaction was monitored by LCMS. The mixture was allowed to cool to room temperature. The residue was purified by reverse-phase flash chromatography using the following conditions: column = spherical C18, 20-40 μm, 330 g, mobile phase A = water (+ 5 mM NH4HCO3), mobile phase B = ACN, flow rate = 80 mL / min, gradient = 5% to 5% B in 10 min, gradient = 45% B to 60% B in 20 min, detector = 220 nm. Fractions containing the desired product were collected at 55% B and concentrated under reduced pressure to give 4-chloro-5-[2-[(2-hydroxyethyl)amino]-4-[2-(trifluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (210 mg, 59.66%) as a yellow solid.

[0299] 4-chloro-5-[2-[(2-hydroxyethyl)amino]-4-[2-(trifluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2,3-dihydropyridazin-3-one To a stirred solution of 4-chloro-5-[2-[(2-hydroxyethyl)amino]-4-[2-(trifluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (200 mg, 0.353 mmol, 1 equiv.) in DCM (5 mL) was added TFA (2 mL) at room temperature. The resulting mixture was stirred at room temperature for 1 hour. The reaction was monitored by LCMS. The mixture was basified to pH 8 with saturated NaHCO3 (aq.). The resulting mixture was concentrated under reduced pressure. The residue was purified by preparative HPLC using the following conditions: (Column: XBridge Prep OBD C18 column, 30 x 150 mm, 5 um, Mobile phase A = unspecified, Mobile phase B = unspecified, Flow rate = 60 mL / min, Gradient = 25% B to 50% B in 8 min, 220 nm, Rt = 7.67 min) to give 4-chloro-5-[2-[(2-hydroxyethyl)amino]-4-[2-(trifluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2,3-dihydropyridazin-3-one (106.3 mg) as a white solid.

[0300] Example 11 Synthesis of Compounds 138 and 139

[0301] [ka] 7-Benzyl-2-chloro-4-[4-fluoro-2-(trifluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine To a stirred solution of 4-fluoro-2-(trifluoromethyl)phenol (1469.32 mg, 8.158 mmol, 1.20 equiv.) and 7-benzyl-2,4-dichloro-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine (2000 mg, 6.799 mmol, 1 equiv.) in DMF (20 mL) was added KCO (1879.20 mg, 13.597 mmol, 2 equiv.) at room temperature. The solution was stirred at 70 °C for 0.5 h. The mixture was concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography using the following conditions: column = spherical C18, 20-40 μm, 330 g, mobile phase A = water (+ 5 mM NTFA), mobile phase B = acetonitrile, flow rate = 80 mL / min, gradient = 5% to 5% B in 10 min, gradient = 70% B to 95% B in 20 min, detector = 254 nm. Fractions containing the desired product were collected at 95% B and concentrated under reduced pressure to give 7-benzyl-2-chloro-4-[4-fluoro-2-(trifluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine (2331 mg, 78.31%) as an off-white solid.

[0302] 7-Benzyl-4-[4-fluoro-2-(trifluoromethyl)phenoxy]-1H,2H,5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-2-one A solution of 7-benzyl-2-chloro-4-[4-fluoro-2-(trifluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine (2 g, 4.568 mmol, 1 equiv.) in HAc (10 mL, 174.515 mmol, 38.20 equiv.) and HO (1 mL, 55.508 mmol, 12.15 equiv.) was stirred at 140 °C for 10 h under a N atmosphere. The resulting mixture was concentrated under reduced pressure. The residue was purified by preparative TLC (PE / EtOAc = 1:1) to give 7-benzyl-4-[4-fluoro-2-(trifluoromethyl)phenoxy]-1H,2H,5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-2-one (530 mg, 27.67%) as a pale yellow solid.

[0303] 4-[4-Fluoro-2-(trifluoromethyl)phenoxy]-1H,2H,5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-2-one To a solution of 7-benzyl-4-[4-fluoro-2-(trifluoromethyl)phenoxy]-1H,2H,5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-2-one (530 mg, 1.264 mmol, 1 equiv.) in MeOH (10 mL, 246.989 mmol, 195.44 equiv.), Pd / C (268.98 mg, 2.528 mmol, 2 equiv.) was added under a nitrogen atmosphere. The mixture was hydrogenated under a hydrogen atmosphere using a hydrogen balloon at room temperature for 4 hours, filtered through a Celite pad, and concentrated under reduced pressure. 4-[4-fluoro-2-(trifluoromethyl)phenoxy]-1H,2H,5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-2-one (430 mg, 103.34%) was obtained as a pale yellow solid.

[0304] 4-chloro-5-[4-[4-fluoro-2-(trifluoromethyl)phenoxy]-2-oxo-1H,2H,5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one A mixture of 4-[4-fluoro-2-(trifluoromethyl)phenoxy]-1H,2H,5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-2-one (430 mg, 1.306 mmol, 1 equiv.) and 4,5-dichloro-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (357.84 mg, 1.437 mmol, 1.1 equiv.) in DIEA (337.58 mg, 2.612 mmol, 2.00 equiv.) was stirred at 100° C. for 2 h under a N atmosphere. The residue was purified by preparative TLC (PE / EA=1:1) to give 4-chloro-5-[4-[4-fluoro-2-(trifluoromethyl)phenoxy]-2-oxo-1H,2H,5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (210 mg, 29.67%) as a pale yellow solid.

[0305] 4-chloro-5-[4-[4-fluoro-2-(trifluoromethyl)phenoxy]-1-methyl-2-oxo-1H,2H,5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one and 4-chloro-5-[4-[4-fluoro-2-(trifluoromethyl)phenoxy]-2-methoxy-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one To a solution of 4-chloro-5-[4-[4-fluoro-2-(trifluoromethyl)phenoxy]-2-oxo-1H,2H,5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (90 mg, 0.166 mmol, 1 equiv.) and NaHCO (27.90 mg, 0.332 mmol, 2 equiv.) in DMF (10 mL, 129.218 mmol, 778.02 equiv.), CHI (47.15 mg, 0.332 mmol, 2.00 equiv.) was added dropwise at 0° C. under a nitrogen atmosphere. The mixture was stirred at 25° C. for 16 h. The resulting mixture was concentrated under reduced pressure. The residue was purified by preparative TLC (PE / EA=0 / 1) to give 4-chloro-5-[4-[4-fluoro-2-(trifluoromethyl)phenoxy]-1-methyl-2-oxo-1H,2H,5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (60 mg, 64.99%) and 4-chloro-5-[4-[4-fluoro-2-(trifluoromethyl)phenoxy]-2-methoxy-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (15 mg) as a light yellow solid.

[0306] 4-chloro-5-[4-[4-fluoro-2-(trifluoromethyl)phenoxy]-1-methyl-2-oxo-1H,2H,5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2,3-dihydropyridazin-3-one To a solution of 4-chloro-5-[4-[4-fluoro-2-(trifluoromethyl)phenoxy]-1-methyl-2-oxo-1H,2H,5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (60 mg, 0.108 mmol, 1 equiv) in DCM (10 mL, 157.300 mmol, 1457.41 equiv) was added TFA (123.07 mg, 1.079 mmol, 10 equiv) at 25° C. The resulting mixture was concentrated under reduced pressure. The crude product (100 mg) was purified by preparative HPLC using the following conditions: (Column: XBridge Shield RP18 OBD column 30 × 150 mm, 5 μm, Mobile phase A = water (10 mM NH4HCO3), Mobile phase B = acetonitrile, Flow rate = 60 mL / min, Gradient = 20% B to 40% B in 7 min, 220 nm, Rt = 6.63 min) to give 4-chloro-5-[4-[4-fluoro-2-(trifluoromethyl)phenoxy]-1-methyl-2-oxo-1H,2H,5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2,3-dihydropyridazin-3-one (29.3 mg, 57.54%) as a white solid.

[0307] 4-chloro-5-[4-[4-fluoro-2-(trifluoromethyl)phenoxy]-2-methoxy-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2,3-dihydropyridazin-3-one To a solution of 4-chloro-5-[4-[4-fluoro-2-(trifluoromethyl)phenoxy]-2-methoxy-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (15 mg, 0.027 mmol, 1 equiv) in DCM (5 mL, 78.650 mmol, 2914.83 equiv) was added TFA (30.77 mg, 0.270 mmol, 10 equiv) at 25° C. The resulting mixture was concentrated under reduced pressure. The crude product (20 mg) was purified by preparative HPLC using the following conditions: (Column: XBridge Shield RP18 OBD column, 30 × 150 mm, 5 μm, Mobile phase A = water (10 mM NH4HCO3), Mobile phase B = acetonitrile, Flow rate = 60 mL / min, Gradient = 20% B to 40% B in 7 min, 220 nm, Rt = 6.63 min) to give 4-chloro-5-[4-[4-fluoro-2-(trifluoromethyl)phenoxy]-2-methoxy-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2,3-dihydropyridazin-3-one (7.5 mg, 58.91%) as a white solid.

[0308] Example 12 Synthesis of Compound 110

[0309] [ka] tert-Butyl 2-chloro-4-[2-(trifluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate To a stirred solution of tert-butyl 2,4-dichloro-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (2 g, 6.58 mmol, 1 equiv.) and 2-(trifluoromethyl)phenol (1.6 g, 9.86 mmol, 1.5 equiv.) in acetonitrile (20 mL) was added DBU (2.0 g, 13.15 mmol, 2 equiv.) at room temperature. The solution was stirred at room temperature for 4 hours. The mixture was concentrated under reduced pressure. The residue was purified by preparative TLC (PE / EtOAc = 10:1) to afford tert-butyl 2-chloro-4-[2-(trifluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (700 mg, 24.77%) as a colorless oil.

[0310] tert-Butyl 2-methoxy-4-[2-(trifluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate To a solution of tert-butyl 2-chloro-4-[2-(trifluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (1 g, 2.327 mmol, 1 equiv.) in MeOH (20 mL, 493.978 mmol, 212.32 equiv.) was added NaOMe (0.25 g, 0.005 mmol, 2 equiv.) at 25° C. The mixture was stirred at 25° C. for 4 hours. The resulting mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with PE / EA (10 / 1 to 1 / 1) to give tert-butyl 2-methoxy-4-[2-(trifluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (100 mg, 10.10%) as a pale yellow solid.

[0311] 2-Methoxy-4-[2-(trifluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine To a solution of tert-butyl 2-methoxy-4-[2-(trifluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (100 mg, 0.235 mmol, 1 equiv) in DCM (10 mL) was added TFA (268.03 mg, 2.351 mmol, 10 equiv) at 25° C. The solution was stirred at 25° C. for 4 h. The resulting mixture was concentrated under reduced pressure. The crude product (150 mg) was purified by preparative HPLC using the following conditions: (Column: XBridge Shield RP18 OBD column, 30 × 150 mm, 5 μm, Mobile phase A = water (10 mM NH4HCO3), Mobile phase B = acetonitrile, Flow rate = 60 mL / min, Gradient = 20% B to 40% B in 7 min, 220 nm, Rt = 6.63 min) to give 2-methoxy-4-[2-(trifluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine (80 mg, 104.62%) as a pale yellow solid.

[0312] 4-chloro-5-[2-methoxy-4-[2-(trifluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one A solution of 2-methoxy-4-[2-(trifluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine (80 mg, 0.246 mmol, 1 equiv.) and 4,5-dichloro-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (61.26 mg, 0.246 mmol, 1 equiv.) in DIEA (63.57 mg, 0.492 mmol, 2.00 equiv.) was stirred at 100° C. for 2 h under a nitrogen atmosphere. The residue was purified by silica gel column chromatography eluting with PE / EA (5 / 1 to 1 / 1) to give 4-chloro-5-[2-methoxy-4-[2-(trifluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (120 mg, 90.71%) as a pale yellow solid.

[0313] 4-chloro-5-[2-methoxy-4-[2-(trifluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2,3-dihydropyridazin-3-one To a solution of 4-chloro-5-[2-methoxy-4-[2-(trifluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (120 mg, 0.223 mmol, 1 equiv) in DCM (5 mL, 78.650 mmol, 352.56 equiv) was added TFA (254.36 mg, 2.231 mmol, 10.00 equiv) at 25° C. The resulting mixture was concentrated under reduced pressure. The crude product (150 mg) was purified by preparative HPLC using the following conditions: (Column: XBridge Shield RP18 OBD column, 30 × 150 mm, 5 μm, Mobile phase A = water (10 mM NH4HCO3), Mobile phase B = acetonitrile, Flow rate = 60 mL / min, Gradient = 20% B to 40% B in 7 min, 220 nm, Rt = 6.63 min) to give 4-chloro-5-[2-methoxy-4-[2-(trifluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2,3-dihydropyridazin-3-one (24.1 mg, 23.81%) as a white solid.

[0314] Example 13 Synthesis of Compound 108

[0315] [ka] tert-Butyl 4-(3-bromo-2-chlorophenoxy)-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate To a stirred solution of tert-butyl 4-chloro-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (500 mg, 1.854 mmol, 1 equiv.) and 3-bromo-2-chlorophenol (461.46 mg, 2.224 mmol, 1.20 equiv.) in DMF (10 mL) was added KCO (512.38 mg, 3.707 mmol, 2 equiv.). The resulting mixture was stirred at 70 °C for 1 h. The mixture was purified by reverse-phase flash chromatography using the following conditions: spherical C18, 20-40 μm, 330 g, mobile phase A = water (5 mM NH4HCO3), mobile phase B = acetonitrile, flow rate = 80 mL / min, gradient = 20% B to 60% B in 55 min, 254 nm. Fractions containing the desired product were collected at 40% B and concentrated under reduced pressure. This gave tert-butyl 4-(3-bromo-2-chlorophenoxy)-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (300 mg, 36.72%) as an off-white solid.

[0316] tert-Butyl 4-(2-chloro-3-cyanophenoxy)-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate To a stirred solution of tert-butyl 4-(3-bromo-2-chlorophenoxy)-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (450 mg, 1.021 mmol, 1 equiv.) and zinc dicarbonitrile (143.87 mg, 1.225 mmol, 1.20 equiv.) in DMF (5 mL) was added Pd(PPh3)4 (117.99 mg, 0.102 mmol, 0.1 equiv.). The resulting mixture was stirred at 120 °C under a nitrogen atmosphere for 2 h. The residue was purified by reverse-phase flash chromatography using the following conditions: column = spherical C18, 20-40 μm, 180 g, mobile phase A = water (5 mM NH4HCO3), mobile phase B = acetonitrile, flow rate = 45 mL / min, gradient = 10% B to 60% B in 55 min, 254 nm. Fractions containing the desired product were collected at 40% B and concentrated under reduced pressure. This afforded tert-butyl 4-(2-chloro-3-cyanophenoxy)-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (280 mg, 70.89%) as a pale yellow solid.

[0317] 2-Chloro-3-[5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-4-yloxy]benzonitrile To a stirred solution of tert-butyl 4-(2-chloro-3-cyanophenoxy)-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (100 mg, 0.259 mmol, 1 equiv.) in DCM (3 mL) was added TFA (1 mL). The resulting mixture was stirred at room temperature under air for 2 hours. The resulting mixture was concentrated under reduced pressure. The mixture was basified to pH 7 with saturated NH4HCO3 (aq.). The mixture was purified by reverse-phase flash chromatography using the following conditions: column = spherical C18, 20-40 μm, 180 g, mobile phase A = water (5 mM NH4HCO3), mobile phase B = acetonitrile, flow rate = 45 mL / min, gradient = 30% B to 60% B in 30 min, 254 nm. Fractions containing the desired product were collected at 45% B and concentrated under reduced pressure. This gave 2-chloro-3-[5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-4-yloxy]benzonitrile (60 mg, 80.95%) as a pale yellow oil.

[0318] 2-chloro-3-([7-[5-chloro-1-(oxan-2-yl)-6-oxo-1,6-dihydropyridazin-4-yl]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-4-yl]oxy)benzonitrile To a stirred solution of tert-butyl 4-(2-chloro-3-cyanophenoxy)-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (60 mg, 0.155 mmol, 1 equiv.) and 4,5-dichloro-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (38.63 mg, 0.155 mmol, 1.00 equiv.) in DIEA (40.09 mg, 0.310 mmol, 2 equiv.) The resulting mixture was stirred at 100° C. under air for hours. The residue was purified by preparative TLC (PE / EtOAc=1:1) to give 2-chloro-3-([7-[5-chloro-1-(oxan-2-yl)-6-oxo-1,6-dihydropyridazin-4-yl]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-4-yl]oxy)benzonitrile (50 mg, 64.56%) as a pale yellow solid.

[0319] 2-chloro-3-[[7-(5-chloro-6-oxo-1,6-dihydropyridazin-4-yl)-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-4-yl]oxy]benzonitrile To a stirred solution of 2-chloro-3-([7-[5-chloro-1-(oxan-2-yl)-6-oxo-1,6-dihydropyridazin-4-yl]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-4-yl]oxy)benzonitrile (50 mg, 0.100 mmol, 1 equiv) in DCM (3 mL) was added TFA (1 mL). The resulting mixture was stirred at room temperature for 2 hours. The resulting mixture was concentrated under reduced pressure. The mixture was basified to pH 7 with saturated NH4CO3 (aq). The crude product was purified by preparative HPLC using the following conditions: (Column: XBridge Prep OBD C18 column, 30 x 150 mm, 5 um, Mobile phase A = water (10 mM NH4HCO3), Mobile phase B = acetonitrile, Flow rate = 60 mL / min, Gradient = 20% B to 42% B in 8 min, 220 nm, Rt = 7.58 min) to give 2-chloro-3-[[7-(5-chloro-6-oxo-1,6-dihydropyridazin-4-yl)-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-4-yl]oxy]benzonitrile (14.5 mg, 34.88%) as an off-white solid.

[0320] Example 14 Synthesis of Compound 111

[0321] [ka] tert-Butyl 4-[3-bromo-2-(trifluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate To a stirred mixture of tert-butyl 4-chloro-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (180 mg, 0.667 mmol, 1 equiv.) and 3-bromo-2-(trifluoromethyl)phenol (241.25 mg, 1.001 mmol, 1.50 equiv.) in DMF (10 mL) was added CsCO (434.86 mg, 1.335 mmol, 2.00 equiv.) in portions at room temperature under a nitrogen atmosphere. The resulting mixture was stirred at 70 °C under a nitrogen atmosphere for 0.5 h. The reaction was monitored by LCMS. The mixture was allowed to cool to room temperature. The resulting mixture was concentrated under reduced pressure. The resulting mixture was extracted with EtOAc (3 × 200 mL). The combined organic layers were washed with brine (2 × 100 mL) and dried over anhydrous NaSO. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography using the following conditions: (column: C18 silica gel, mobile phase: aqueous acetonitrile, gradient from 40% to 85% in 30 min, detector: UV 220 nm) to give tert-butyl 4-[3-bromo-2-(trifluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (150 mg, 47.39%) as a yellow oil.

[0322] tert-Butyl 4-[3-cyano-2-(trifluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate To a stirred mixture of tert-butyl 4-[3-bromo-2-(trifluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (150 mg, 0.316 mmol, 1 equiv.) and Zn(CN) (111.43 mg, 0.949 mmol, 3.00 equiv.) in DMF (8 mL), Pd(PPh) (36.55 mg, 0.032 mmol, 0.1 equiv.) was added in small portions at room temperature under a nitrogen atmosphere. The final reaction mixture was irradiated by microwave irradiation at 150 °C for 3 h. The reaction was monitored by LCMS. The residue was purified by reverse-phase flash chromatography using the following conditions: (column = C18 silica gel, mobile phase = aqueous acetonitrile, 40% to 95% gradient in 30 min, detector = UV 220 nm) to give tert-butyl 4-[3-cyano-2-(trifluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (70 mg, 52.65%) as a yellow oil.

[0323] 3-[5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-4-yloxy]-2-(trifluoromethyl)benzonitrile To a stirred solution of tert-butyl 4-[3-cyano-2-(trifluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (70 mg) in DCM (10 mL) was added dropwise TFA (1 mL) at room temperature. The reaction mixture was stirred at room temperature for 2 hours. The reaction was monitored by LCMS. The resulting mixture was concentrated under reduced pressure. The residue was basified to pH = 8 with saturated NH4HCO3 (aq). The resulting mixture was extracted with DCM (3 x 100 mL). The combined organic layers were washed with brine (1 x 100 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography using the following conditions: (column: C18 silica gel, mobile phase: aqueous acetonitrile, gradient from 30% to 60% in 20 min, detector: UV 220 nm) to give 3-[5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-4-yloxy]-2-(trifluoromethyl)benzonitrile (40 mg) as a yellow oil.

[0324] 3-([7-[5-chloro-1-(oxan-2-yl)-6-oxo-1,6-dihydropyridazin-4-yl]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-4-yl]oxy)-2-(trifluoromethyl)benzonitrile To a 25 mL round-bottom flask was added 3-[5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-4-yloxy]-2-(trifluoromethyl)benzonitrile (40 mg, 0.125 mmol, 1 equiv.), 4,5-dichloro-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (62.22 mg, 0.250 mmol, 2.00 equiv.), and DIEA (48.42 mg, 0.375 mmol, 3.00 equiv.) under a nitrogen atmosphere at room temperature. The resulting mixture was stirred at 90° C. for 16 hours under a nitrogen atmosphere. The residue was purified by preparative TLC (PE / EtOAc=5 / 1) to give 3-([7-[5-chloro-1-(oxan-2-yl)-6-oxo-1,6-dihydropyridazin-4-yl]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-4-yl]oxy)-2-(trifluoromethyl)benzonitrile (50 mg, 75.12%) as a yellow oil.

[0325] 3-[[7-(5-chloro-6-oxo-1,6-dihydropyridazin-4-yl)-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-4-yl]oxy]-2-(trifluoromethyl)benzonitrile To a stirred solution of 3-([7-[5-chloro-1-(oxan-2-yl)-6-oxo-1,6-dihydropyridazin-4-yl]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-4-yl]oxy)-2-(trifluoromethyl)benzonitrile (50 mg) in DCM (10 mL) was added TFA (1 mL) dropwise at room temperature. The reaction mixture was stirred at room temperature for 2 hours. The reaction was monitored by LCMS. The resulting mixture was concentrated under reduced pressure. The residue was basified to pH = 8 with saturated NH4HCO3 (aq). The resulting mixture was extracted with DCM (3 x 100 mL). The combined organic layers were washed with brine (1 x 100 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative HPLC using the following conditions: (Column = XBridge Prep OBD C18 column 30 x 150 mm, 5 um, Mobile phase A = water (10 mM NH4HCO3), Mobile phase B = acetonitrile, Flow rate = 60 mL / min, Gradient = 25% B to 45% B in 8 min, 220 nm, Rt = 7.07 min) to give 3-[[7-(5-chloro-6-oxo-1,6-dihydropyridazin-4-yl)-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-4-yl]oxy]-2-(trifluoromethyl)benzonitrile (10.8 mg) as a white solid.

[0326] Example 15 Synthesis of Compounds 126 and 126a

[0327] [ka] N-[(1E)-1-[4-fluoro-2-(trifluoromethyl)phenyl]ethylidene]-4-methylbenzene-1-sulfonohydrazide To a stirred solution of 1-[4-fluoro-2-(trifluoromethyl)phenyl]ethan-1-one (2 g, 9.702 mmol, 1 equiv.) in EtOH (40 mL) was added 4-methylbenzene-1-sulfonohydrazide (1.81 g, 9.719 mmol, 1.00 equiv.) in small portions at room temperature under a nitrogen atmosphere. The resulting mixture was stirred at 90 °C for 6 hours under a nitrogen atmosphere. The reaction was monitored by LCMS. The mixture was allowed to cool to room temperature. The resulting mixture was concentrated in vacuo. The residue was purified by reverse-phase flash chromatography using the following conditions: column = spherical C18, 20-40 μm, 330 g, mobile phase A = water (+ 5 mM AcOH), mobile phase B = acetonitrile, flow rate = 80 mL / min, gradient = 5% B to 5% B in 10 min, 45% B to 70% B in 20 min, detector = 220 nm. Fractions containing the desired product were collected at 60% B and concentrated under reduced pressure to give N-[(1E)-1-[4-fluoro-2-(trifluoromethyl)phenyl]ethylidene]-4-methylbenzene-1-sulfonohydrazide (2.5 g, 68.83%) as a white solid.

[0328] tert-Butyl 4-[1-[4-fluoro-2-(trifluoromethyl)phenyl]ethenyl]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate To a stirred mixture of tert-butyl 4-chloro-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (750 mg, 2.781 mmol, 1 equiv.) and N-[(1E)-1-[4-fluoro-2-(trifluoromethyl)phenyl]ethylidene]-4-methylbenzene-1-sulfonohydrazide (2081.80 mg, 5.561 mmol, 2.00 equiv.) in 1,4-dioxane (20 mL), Pd(acetonitrile)Cl (72.14 mg, 0.278 mmol, 0.10 equiv.), Dppf (307.18 mg, 0.556 mmol, 0.2 equiv.), and t-BuOLi (489.71 mg, 6.117 mmol, 2.20 equiv.) were added in small portions at room temperature under a nitrogen atmosphere. The final reaction mixture was irradiated at 100°C for 2 hours using microwave irradiation. The reaction was monitored by LCMS. The mixture was allowed to cool to room temperature. The resulting mixture was filtered, and the filter cake was washed with EtOAc (2 x 50 mL). The filtrate was concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography using the following conditions: Column = spherical C18, 20-40 μm, 330 g, Mobile Phase A = water (+ 5 mM AcOH), Mobile Phase B = acetonitrile, Flow Rate = 80 mL / min, Gradient = 5% to 5% B in 10 min, 50% B to 90% B in 30 min, Detector = 220 nm. Fractions containing the desired product were collected at 85% B and concentrated under reduced pressure to give tert-butyl 4-[1-[4-fluoro-2-(trifluoromethyl)phenyl]ethenyl]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (800 mg, 67.95%) as a brown oil.

[0329] tert-Butyl 4-[1-[4-fluoro-2-(trifluoromethyl)phenyl]ethyl]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate To a solution of tert-butyl 4-[1-[4-fluoro-2-(trifluoromethyl)phenyl]ethenyl]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (150 mg) in 30 mL of MeOH was added Pd / C (10%, 30 mg) under a nitrogen atmosphere in a 100 mL round-bottom flask. The mixture was hydrogenated under a hydrogen atmosphere using a hydrogen balloon at room temperature for 4 hours, filtered through a Celite pad, and concentrated under reduced pressure to give tert-butyl 4-[1-[4-fluoro-2-(trifluoromethyl)phenyl]ethyl]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (150 mg) as a yellow oil.

[0330] 4-[1-[4-fluoro-2-(trifluoromethyl)phenyl]ethyl]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine To a stirred solution of tert-butyl 4-[1-[4-fluoro-2-(trifluoromethyl)phenyl]ethyl]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (150 mg) in DCM (10 mL) was added dropwise TFA (1 mL) at room temperature. The reaction mixture was stirred at room temperature for 2 hours. The reaction was monitored by LCMS. The resulting mixture was concentrated under reduced pressure. The residue was basified to pH = 8 with saturated NH4HCO3 (aq). The resulting mixture was extracted with DCM (3 x 50 mL). The combined organic layers were washed with brine (1 x 30 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography using the following conditions: column = spherical C18, 20-40 μm, 120 g, mobile phase A = water (+ 5 mM AcOH), mobile phase B = acetonitrile, flow rate = 45 mL / min, gradient = 5% to 5% B in 10 min, 40% B to 58% B in 15 min, detector = 254 nm. Fractions containing the desired product were collected at 53% B and concentrated under reduced pressure to give 4-[1-[4-fluoro-2-(trifluoromethyl)phenyl]ethyl]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine (100 mg) as a yellow oil.

[0331] 4-chloro-5-(4-[1-[4-fluoro-2-(trifluoromethyl)phenyl]ethyl]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl)-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one To a 50 mL round-bottom flask, 4-[1-[4-fluoro-2-(trifluoromethyl)phenyl]ethyl]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine (100 mg, 0.307 mmol, 1 equiv.), 4,5-dichloro-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (91.88 mg, 0.369 mmol, 1.20 equiv.), and DIEA (119.19 mg, 0.922 mmol, 3.00 equiv.) were added under a nitrogen atmosphere at room temperature. The resulting mixture was stirred at 90° C. under a nitrogen atmosphere for 2 hours. The reaction was monitored by LCMS. The mixture was allowed to cool to room temperature. The residue was purified by reverse-phase flash chromatography using the following conditions: column = spherical C18, 20-40 μm, 120 g, mobile phase A = water (+ 5 mM AcOH), mobile phase B = acetonitrile, flow rate = 45 mL / min, gradient = 5% to 5% B in 10 min, gradient = 40% B to 60% B in 15 min, detector = 220 nm. Fractions containing the desired product were collected at 53% B and concentrated under reduced pressure to give 4-chloro-5-(4-[1-[4-fluoro-2-(trifluoromethyl)phenyl]ethyl]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl)-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (120 mg, 72.57%) as a yellow oil.

[0332] 4-chloro-5-[4-[(1S)-1-[4-fluoro-2-(trifluoromethyl)phenyl]ethyl]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2,3-dihydropyridazin-3-one and 4-chloro-5-[4-[(1R)-1-[4-fluoro-2-(trifluoromethyl)phenyl]ethyl]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2,3-dihydropyridazin-3-one To a stirred solution of 4-chloro-5-(4-[1-[4-fluoro-2-(trifluoromethyl)phenyl]ethyl]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl)-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (200 mg) in DCM (10 mL) was added TFA (1 mL) dropwise at room temperature. The reaction mixture was stirred at room temperature for 4 hours. The reaction was monitored by LCMS. The resulting mixture was concentrated under reduced pressure. The residue was basified to pH = 8 with saturated NH4HCO3 (aq). The resulting mixture was extracted with DCM (3 x 100 mL). The combined organic layers were washed with brine (1 x 100 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by chiral preparative HPLC using the following conditions: (Column: XBridge Preparative Phenyl OBD Column 19 x 150 mm, 5 μm, 13 nm, Mobile Phase A:Mobile Phase B = Flow Rate = 60 mL / min, Gradient = 20% B to 37% B in 8 min, 220 nm, Rt = 7.97 min). Two isomers were separated by this technique, but absolute orientation was not determined. The compound designated 4-chloro-5-[4-[(1S)-1-[4-fluoro-2-(trifluoromethyl)phenyl]ethyl]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2,3-dihydropyridazin-3-one (11.8 mg) was obtained in 1.819 min as an off-white solid. The compound designated 4-chloro-5-[4-[(1R)-1-[4-fluoro-2-(trifluoromethyl)phenyl]ethyl]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2,3-dihydropyridazin-3-one (13.5 mg) was obtained as a white solid in 2.470 minutes.

[0333] Example 16 Synthesis of Compound 133

[0334] [ka] tert-Butyl 4-[methyl[(3R,4R)-4-methylpiperidin-3-yl]amino]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate To a 25 mL round-bottom flask were added (3R,4R)-1-benzyl-N,4-dimethylpiperidin-3-amine (2.43 g, 0.011 mmol, 1.50 equiv.) and tert-butyl 4-chloro-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (2 g, 0.007 mmol, 1 equiv.) at room temperature. To the mixture was added DIEA (1.92 g, 0.015 mmol, 2.00 equiv.) at room temperature. The mixture was stirred at 100° C. for 2 hours. The residue was purified by preparative TLC (PE / EtOAc=1:1) to give tert-butyl 4-[methyl[(3R,4R)-4-methylpiperidin-3-yl]amino]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (670 mg, 25.00%) as an off-white solid.

[0335] (3R,4R)-1-Benzyl-N,4-dimethyl-N-[5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-4-yl]piperidin-3-amine To a stirred solution of tert-butyl 4-[[(3R,4R)-1-benzyl-4-methylpiperidin-3-yl](methyl)amino]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (413 mg, 0.914 mmol, 1 equiv.) in DCM (10 mL) was added trifluoroacetic acid (3 mL, 0.026 mmol, 6.00 equiv.) dropwise at 0° C. The mixture was stirred at room temperature for 2 hours. The reaction was monitored by LCMS. The solution was concentrated under reduced pressure. The crude product (362 mg) was purified by preparative HPLC using the following conditions: (Column = XBridge Shield RP18 OBD column, 5 μm, 19 × 150 mm, Mobile phase A = water (10 mM NH4HCO3), Mobile phase B = acetonitrile, Flow rate = 80 mL / min, Gradient = 30% B to 80% B in 25 min, 220 nm, Rt = 21.65 min) to afford (3R,4R)-1-benzyl-N,4-dimethyl-N-[5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-4-yl]piperidin-3-amine (250 mg, 77.77%) as a red oil.

[0336] 5-(4-[[(3R,4R)-1-benzyl-4-methylpiperidin-3-yl](methyl)amino]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl)-4-chloro-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one To a 25 mL round-bottom flask were added (3R,4R)-1-benzyl-N,4-dimethyl-N-[5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-4-yl]piperidin-3-amine (263 mg, 0.748 mmol, 1 equivalent) and 4,5-dichloro-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (186.38 mg, 0.748 mmol, 1.00 equivalent) at room temperature. DIEA (193.41 mg, 1.261 mmol, 2 equivalents) was added to the mixture at room temperature. The mixture was stirred at 100° C. for 2 hours. The residue was purified by reverse-phase flash chromatography using the following conditions: column = spherical C18, 20-40 μm, 330 g, mobile phase A = water (+ 5 mM NH4HCO3), mobile phase B = acetonitrile, flow rate = 80 mL / min, gradient = 5% to 5% B in 10 min, 45% to 95% B in 30 min, detector = 254 nm. Fractions containing the desired product were collected at 85% B and concentrated under reduced pressure to give 5-(4-[[(3R,4R)-1-benzyl-4-methylpiperidin-3-yl](methyl)amino]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl)-4-chloro-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (245 mg, 58.04%) as an off-white solid.

[0337] 5-(4-[[(3R,4R)-1-benzyl-4-methylpiperidin-3-yl](methyl)amino]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl)-4-chloro-2,3-dihydropyridazin-3-one To a stirred solution of 5-(4-[[(3R,4R)-1-benzyl-4-methylpiperidin-3-yl](methyl)amino]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl)-4-chloro-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (88 mg, 1 equiv.) in DCM (10 mL) was added trifluoroacetic acid (3 mL, 0.026 mmol, 6.00 equiv.) dropwise at 0° C. The mixture was stirred at room temperature for 2 hours. The solution was concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography using the following conditions: column = spherical C18, 20-40 μm, 330 g, mobile phase A = water (+ 5 mM NTFA), mobile phase B = acetonitrile, flow rate = 80 mL / min, gradient = 5% to 5% B in 10 min, gradient = 33% to 95% B in 30 min, detector = 254 nm. Fractions containing the desired product were collected at 90% B and concentrated under reduced pressure to give 5-(4-[[(3R,4R)-1-benzyl-4-methylpiperidin-3-yl](methyl)amino]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl)-4-chloro-2,3-dihydropyridazin-3-one (33.5 mg, 44.74%) as an off-white solid.

[0338] Compound 133a was prepared by the methods and schemes described in this example by using (3S,4S)-1-benzyl-N,4-dimethylpiperidin-3-amine instead of (3R,4R)-1-benzyl-N,4-dimethylpiperidin-3-amine.

[0339] Example 17 Synthesis of Compound 136

[0340] [ka] tert-Butyl 4-[[4-fluoro-2-(trifluoromethyl)phenyl]amino]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate A mixture of 4-fluoro-2-(trifluoromethyl)aniline (6.64 g, 37.074 mmol, 2 equiv.), tert-butyl 4-chloro-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (5 g, 18.537 mmol, 1 equiv.), Pd(AcO) (0.83 g, 3.707 mmol, 0.2 equiv.), Xantphos (4.29 g, 7.415 mmol, 0.4 equiv.), and CsCO (12.08 g, 37.074 mmol, 2 equiv.) in 1,4-dioxane (80 mL) was stirred at 110 °C for 16 h. The reaction mixture was filtered, and the filtrate was concentrated to give the crude product, which was purified by silica gel column chromatography eluting with PE:EA (20:1 to 1:2) to give tert-butyl 4-[[4-fluoro-2-(trifluoromethyl)phenyl]amino]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (5.6 g, 73.26%) as a white solid.

[0341] tert-Butyl 4-[[4-fluoro-2-(trifluoromethyl)phenyl](2-methoxy-2-oxoethyl)amino]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate To a stirred mixture of tert-butyl 4-[[4-fluoro-2-(trifluoromethyl)phenyl]amino]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (3 g, 7.275 mmol, 1 equiv.) and methyl 2-bromoacetate (2.23 g, 14.578 mmol, 2.00 equiv.) in DMF (30 mL) was added CsCO (4.74 g, 14.548 mmol, 2.00 equiv.) in portions at room temperature under a nitrogen atmosphere. The resulting mixture was stirred at room temperature for 2 hours. The reaction was monitored by LCMS. The resulting mixture was extracted with EtOAc (3 × 400 mL). The combined organic layers were washed with brine (2 × 200 mL) and dried over anhydrous NaSO. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography using the following conditions: column = spherical C18, 20-40 μm, 330 g, mobile phase A = water (+ 5 mM TFA), mobile phase B = acetonitrile, flow rate = 80 mL / min, gradient = 5% to 5% B in 10 min, gradient = 55% to 85% B in 30 min, detector = 220 nm. Fractions containing the desired product were collected at 79% B and concentrated under reduced pressure to give tert-butyl 4-[[4-fluoro-2-(trifluoromethyl)phenyl](2-methoxy-2-oxoethyl)amino]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (500 mg, 14.19%) as a yellow solid.

[0342] tert-Butyl 4-[[4-fluoro-2-(trifluoromethyl)phenyl](2-hydroxyethyl)amino]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate To a stirred solution of tert-butyl 4-[[4-fluoro-2-(trifluoromethyl)phenyl](2-methoxy-2-oxoethyl)amino]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (500 mg, 1.032 mmol, 1 equiv.) in THF (50 mL) was added LiAlH4 (78.34 mg, 2.064 mmol, 2.00 equiv.) in portions at −30° C. under a nitrogen atmosphere. The reaction mixture was stirred at room temperature for 16 hours. The reaction was monitored by LCMS. The reaction was quenched by the addition of water (1 mL) at −30° C. The precipitated solid was collected by filtration and washed with MeOH (3×30 mL). The resulting mixture was concentrated in vacuo. The residue was purified by preparative TLC (PE / EtOAc=1:1) to give tert-butyl 4-[[4-fluoro-2-(trifluoromethyl)phenyl](2-hydroxyethyl)amino]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (100 mg, 21.23%) as a yellow oil.

[0343] 4-[1-[4-fluoro-2-(trifluoromethyl)phenyl]ethyl]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine To a stirred solution of tert-butyl 4-[1-[4-fluoro-2-(trifluoromethyl)phenyl]ethyl]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (150 mg) in DCM (10 mL) was added dropwise TFA (1 mL) at room temperature. The reaction mixture was stirred at room temperature for 2 hours. The reaction was monitored by LCMS. The resulting mixture was concentrated under reduced pressure. The residue was basified to pH = 8 with saturated NH4HCO3 (aq). The resulting mixture was extracted with DCM (3 x 50 mL). The combined organic layers were washed with brine (1 x 30 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography using the following conditions: column = spherical C18, 20-40 μm, 120 g, mobile phase A = water (+ 5 mM NH4HCO3), mobile phase B = acetonitrile, flow rate = 80 mL / min, gradient = 5% to 5% B in 10 min, 40% B to 58% B in 15 min, detector = 220 nm. Fractions containing the desired product were collected at 53% B and concentrated under reduced pressure to give 4-[1-[4-fluoro-2-(trifluoromethyl)phenyl]ethyl]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine (100 mg) as a yellow oil.

[0344] 4-chloro-5-(4-[[4-fluoro-2-(trifluoromethyl)phenyl](2-hydroxyethyl)amino]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl)-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one To a 50 mL round-bottom flask, 2-[[4-fluoro-2-(trifluoromethyl)phenyl]([5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-4-yl])amino]ethan-1-ol (40 mg, 0.112 mmol, 1 equiv.), 4,5-dichloro-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (55.92 mg, 0.224 mmol, 2.00 equiv.), and DIEA (43.53 mg, 0.337 mmol, 3.00 equiv.) were added under a nitrogen atmosphere at room temperature. The resulting mixture was stirred at 90° C. under a nitrogen atmosphere for 2 hours. The reaction was monitored by LCMS. The mixture was allowed to cool to room temperature. The residue was purified by reverse-phase flash chromatography using the following conditions: column = spherical C18, 20-40 μm, 120 g, mobile phase A = water (+ 5 mM NH4HCO3), mobile phase B = acetonitrile, flow rate = 45 mL / min, gradient = 5% to 5% B in 10 min, gradient = 40% B to 60% B in 15 min, detector = 220 nm. Fractions containing the desired product were collected at 55% B and concentrated under reduced pressure to give 4-chloro-5-(4-[[4-fluoro-2-(trifluoromethyl)phenyl](2-hydroxyethyl)amino]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl)-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (50 mg, 78.28%) as a yellow oil.

[0345] 4-chloro-5-(4-[[4-fluoro-2-(trifluoromethyl)phenyl](2-hydroxyethyl)amino]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl)-2,3-dihydropyridazin-3-one To a stirred solution of 4-chloro-5-(4-[[4-fluoro-2-(trifluoromethyl)phenyl](2-hydroxyethyl)amino]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl)-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (50 mg) in DCM (10 mL) was added TFA (1 mL) dropwise at room temperature. The reaction mixture was stirred at room temperature for 2 hours. The reaction was monitored by LCMS. The resulting mixture was concentrated under reduced pressure. The residue was basified to pH = 8 with saturated NH4HCO3 (aq). The resulting mixture was extracted with EtOAc (2 x 50 mL). The combined organic layers were washed with brine (1 x 100 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative HPLC using the following conditions: (Column: XBridge Prep OBD C18 column, 30 x 150 mm, 5 um, Mobile phase A = unspecified, Mobile phase B = unspecified, Flow rate = 60 mL / min, Gradient = 30% B to 45% B in 8 min, 220 nm, Rt = 7.6 min) to give 4-chloro-5-(4-[[4-fluoro-2-(trifluoromethyl)phenyl](2-hydroxyethyl)amino]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl)-2,3-dihydropyridazin-3-one (6.2 mg) as a white solid.

[0346] Example 18 Synthesis of Compound 132

[0347] [ka] tert-Butyl 4-[1-[4-fluoro-2-(trifluoromethyl)phenyl]cyclopropyl]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate To a stirred solution of t-BuONa (226.97 mg, 2.362 mmol, 2.00 equiv.) in DMSO (20 mL) was added MeSiI (472.57 mg, 2.362 mmol, 2.00 equiv.) in portions at 40° C. under a nitrogen atmosphere. The resulting mixture was stirred at 40° C. under a nitrogen atmosphere for 0.5 h. Then, a solution of tert-butyl 4-[1-[4-fluoro-2-(trifluoromethyl)phenyl]ethenyl]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (500 mg, 1.181 mmol, 1 equiv.) in DMSO (5 mL) was added dropwise at room temperature under a nitrogen atmosphere. The resulting mixture was stirred at room temperature for 1 h under a nitrogen atmosphere. The reaction was monitored by LCMS. The mixture was allowed to cool to room temperature. The resulting mixture was extracted with EtOAc (3×100 mL). The combined organic layers were washed with brine (2 × 100 mL) and dried over anhydrous NaSO. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography using the following conditions: column = spherical C18, 20-40 μm, 330 g, mobile phase A = water (+ 5 mM NHHCO), mobile phase B = acetonitrile, flow rate = 80 mL / min, gradient = 5% to 5% B in 10 min, 55% to 80% B in 25 min, detector = 220 nm. Fractions containing the desired product were collected at 73% B and concentrated under reduced pressure to give tert-butyl 4-[1-[4-fluoro-2-(trifluoromethyl)phenyl]cyclopropyl]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (240 mg, 46.46%) as a yellow oil.

[0348] 4-[1-[4-fluoro-2-(trifluoromethyl)phenyl]cyclopropyl]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine To a stirred solution of tert-butyl 4-[1-[4-fluoro-2-(trifluoromethyl)phenyl]cyclopropyl]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (240 mg, 0.549 mmol, 1 equiv.) in DCM (10 mL) was added TFA (1 mL, 13.463 mmol, 24.54 equiv.) dropwise at room temperature. The reaction mixture was stirred at room temperature for 2 hours. The reaction was monitored by LCMS. The resulting mixture was concentrated under reduced pressure. The residue was basified to pH = 8 with saturated NH4HCO3 (aq.). The resulting mixture was extracted with EtOAc (2 x 100 mL). The combined organic layers were washed with brine (1 x 100 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography using the following conditions: column = spherical C18, 20-40 μm, 120 g, mobile phase A = water (+ 5 mM AcOH), mobile phase B = acetonitrile, flow rate = 45 mL / min, gradient = 5% to 5% B in 10 min, gradient = 33% B to 45% B in 20 min, detector = 254 nm. Fractions containing the desired product were collected at 40% B and concentrated under reduced pressure to give 4-[1-[4-fluoro-2-(trifluoromethyl)phenyl]cyclopropyl]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine (150 mg, 81.05%) as a yellow oil.

[0349] 4-chloro-5-(4-[1-[4-fluoro-2-(trifluoromethyl)phenyl]cyclopropyl]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl)-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one To a 50 mL round-bottom flask, 4-[1-[4-fluoro-2-(trifluoromethyl)phenyl]cyclopropyl]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine (150 mg, 0.445 mmol, 1 equiv.), 4,5-dichloro-2-(oxan-2-yl)-1,2,3,6-tetrahydropyridazin-3-one (134.00 mg, 0.534 mmol, 1.20 equiv.), and DIEA (172.42 mg, 1.334 mmol, 3.00 equiv.) were added under a nitrogen atmosphere at room temperature. The resulting mixture was stirred at 90° C. under a nitrogen atmosphere for 2 hours. The reaction was monitored by LCMS. The mixture was allowed to cool to room temperature. The residue was purified by reverse-phase flash chromatography using the following conditions: column = spherical C18, 20-40 μm, 330 g, mobile phase A = water (+ 5 mM NH4HCO3), mobile phase B = acetonitrile, flow rate = 80 mL / min, gradient = 5% to 5% B in 10 min, 40% B to 60% B in 15 min, detector = 220 nm. Fractions containing the desired product were collected at 54% B and concentrated under reduced pressure to give 4-chloro-5-(4-[1-[4-fluoro-2-(trifluoromethyl)phenyl]cyclopropyl]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl)-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (200 mg, 81.78%) as a yellow oil.

[0350] 4-chloro-5-(4-[1-[4-fluoro-2-(trifluoromethyl)phenyl]cyclopropyl]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl)-2,3-dihydropyridazin-3-one To a stirred solution of 4-chloro-5-(4-[1-[4-fluoro-2-(trifluoromethyl)phenyl]cyclopropyl]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl)-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (200 mg) in DCM (10 mL) was added TFA (2 mL) dropwise at room temperature. The reaction mixture was stirred at room temperature for 2 hours. The reaction was monitored by LCMS. The resulting mixture was concentrated under reduced pressure. The residue was basified to pH = 8 with saturated NH4HCO3 (aq). The resulting mixture was extracted with EtOAc (2 x 100 mL). The combined organic layers were washed with brine (1 x 100 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative HPLC using the following conditions: (Column: XBridge Prep OBD C18 column, 30 x 150 mm, 5 μm, Mobile phase A: unspecified, Mobile phase B: unspecified, Flow rate: 60 mL / min, Gradient: 30% B to 55% B in 8 min, 220 nm, Rt: 7.232 min) to afford 4-chloro-5-(4-[1-[4-fluoro-2-(trifluoromethyl)phenyl]cyclopropyl]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl)-2,3-dihydropyridazin-3-one (39.2 mg) as an off-white solid.

[0351] Example 19 Synthesis of Compound 109

[0352] [ka] tert-Butyl 4-(2-bromo-3-fluorophenoxy)-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate To a stirred solution of tert-butyl 4-chloro-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (500 mg, 1.854 mmol, 1 equiv.) and 2-bromo-3-fluorophenol (424.87 mg, 2.224 mmol, 1.20 equiv.) in DMF (10 mL) was added K2CO3 (512.38 mg, 3.707 mmol, 2 equiv.). The resulting mixture was stirred at 70 °C for 0.5 h. The mixture was allowed to cool to room temperature. The reaction was quenched with water at room temperature. The resulting mixture was extracted with EtOAc (2 × 100 mL). The combined organic layers were washed with brine (2 × 100 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with PE / EtOAc (5:1) to give tert-butyl 4-(2-bromo-3-fluorophenoxy)-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (500 mg, 63.58%) as a white solid.

[0353] tert-Butyl 4-(2-ethenyl-3-fluorophenoxy)-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate To a solution of tert-butyl 4-(2-bromo-3-fluorophenoxy)-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (500 mg, 1.178 mmol, 1 equiv.) and pentamethyl-1,3,2-dioxaborolane (334.72 mg, 2.357 mmol, 2.00 equiv.) in HO (2 mL) and 1,4-dioxane (16 mL) was added KCO (325.75 mg, 2.357 mmol, 2 equiv.) and Pd(PPh) (68.09 mg, 0.059 mmol, 0.05 equiv.). After stirring overnight at 90 °C under a nitrogen atmosphere, the resulting mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with PE / EtOAc (5:1) to give tert-butyl 4-(2-ethenyl-3-fluorophenoxy)-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (250 mg, 57.12%) as a yellow oil.

[0354] tert-Butyl 4-(2-ethyl-3-fluorophenoxy)-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate To a stirred solution of tert-butyl 4-(2-ethenyl-3-fluorophenoxy)-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (250 mg, 0.673 mmol, 1 equiv.) in MeOH (10 mL) was added Pd / C (100 mg, 0.940 mmol, 1.40 equiv.). The resulting mixture was stirred under a hydrogen atmosphere at room temperature for 2 hours. The resulting mixture was filtered, and the filter cake was washed with MeOH (2 × 10 mL). The filtrate was concentrated under reduced pressure. The resulting mixture was concentrated under reduced pressure. This afforded tert-butyl 4-(2-ethyl-3-fluorophenoxy)-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (210 mg, 0.08%) as a black oil.

[0355] 4-(2-ethyl-3-fluorophenoxy)-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine To a stirred solution of tert-butyl 4-(2-ethyl-3-fluorophenoxy)-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (210 mg, 0.562 mmol, 1 equiv.) in DCM (3 mL) was added TFA (1 mL). The resulting mixture was stirred at room temperature under air for 2 hours. The resulting mixture was concentrated under reduced pressure. The mixture was basified to pH 8 with saturated NH4HCO3 (aq.). The mixture was purified by reverse-phase flash chromatography using the following conditions: (Column = spherical C18, 20-40 μm, 180 g, Mobile Phase A = water (5 mM NH4HCO3), Mobile Phase B = acetonitrile, Flow Rate = 45 mL / min, Gradient = 25% B to 60% B in 40 min, 254 nm). Fractions containing the desired product were collected at 40% B and concentrated under reduced pressure. This gave 4-(2-ethyl-3-fluorophenoxy)-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine (120 mg, 78.07%) as a pale yellow oil.

[0356] 4-chloro-5-[4-(2-ethyl-3-fluorophenoxy)-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one To a stirred solution of 4-(2-ethyl-3-fluorophenoxy)-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine (120 mg, 0.439 mmol, 1 equiv.) and 4,5-dichloro-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (109.37 mg, 0.439 mmol, 1.00 equiv.) in DIEA (113.49 mg, 0.878 mmol, 2 equiv.) The resulting mixture was stirred at 100° C. for 2 h under an air atmosphere. The residue was purified by preparative TLC (PE / EtOAc=1:1) to give 4-chloro-5-[4-(2-ethyl-3-fluorophenoxy)-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (100 mg, 46.87%) as a pale yellow solid.

[0357] 4-chloro-5-[4-(2-ethyl-3-fluorophenoxy)-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2,3-dihydropyridazin-3-one To a stirred solution of 4-chloro-5-[4-(2-ethyl-3-fluorophenoxy)-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (100 mg, 0.206 mmol, 1 equiv.) in DCM (3 mL) was added TFA (1 mL). The resulting mixture was stirred at room temperature for 2 hours. The resulting mixture was concentrated under reduced pressure. The mixture was basified to pH 7 with saturated NH4HCO3 (aq.). The crude product was purified by preparative HPLC using the following conditions: (Column = XBridge Prep OBD C18 column, 30 × 150 mm, 5 μm, Mobile phase A = water (10 mM NH4HCO3), Mobile phase B = acetonitrile, Flow rate = 60 mL / min, Gradient = 30% B to 50% B in 8 min, 220 nm, Rt = 7.27 min) to give 4-chloro-5-[4-(2-ethyl-3-fluorophenoxy)-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2,3-dihydropyridazin-3-one (41.6 mg, 50.31%) as a white solid.

[0358] Example 20 Synthesis of Compound 127

[0359] [ka] Methyl 3-(methylamino)pyridine-4-carboxylate To a solution of 3-(methylamino)pyridine-4-carboxylic acid (11 g, 72.296 mmol, 1 equiv) in MeOH (500 mL, 12349.455 mmol, 170.82 equiv) was added SOCl2 (43.01 g, 361.478 mmol, 5 equiv) dropwise at 0 °C. The resulting mixture was stirred at 70 °C for 30 h. The reaction was monitored by LCMS. The mixture was allowed to cool to room temperature. The resulting mixture was concentrated under reduced pressure. The residue was dissolved in ethyl acetate (50 mL). The mixture was basified to pH 8 with saturated NaHCO3 (aq). The resulting mixture was extracted with EtOAc (2 × 20 mL). The combined organic layers were washed with brine (1 × 30 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure to give methyl 3-(methylamino)pyridine-4-carboxylate (9 g, crude) as a yellow solid.

[0360] Methyl 3-(N-methylacetamido)pyridine-4-carboxylate To a stirred solution of methyl 3-(methylamino)pyridine-4-carboxylate (9 g, 54.158 mmol, 1 equiv.) in DCM (100 mL) was added pyridine (21.42 g, 270.791 mmol, 5 equiv.) and acetyl chloride (6.38 g, 81.237 mmol, 1.5 equiv.) dropwise at room temperature. The resulting mixture was stirred at room temperature for 2 h. The reaction was monitored by LCMS. The solution was basified to pH 8 with saturated NaHCO3 (aq.). The resulting mixture was concentrated under reduced pressure. The residue was purified by reverse-phase flash using the following conditions: (Column = spherical C18 column, 330 g, Mobile phase A = water (10 mM NH4HCO3), Mobile phase B = acetonitrile, Flow rate = 80 mL / min, Gradient = 10% B to 30% B in 25 min, 254 / 220 nm) to give methyl 3-(N-methylacetamido)pyridine-4-carboxylate (8 g, 70.94%) as a brown liquid.

[0361] 4-Hydroxy-1-methyl-1,2-dihydro-1,7-naphthyridin-2-one To a stirred solution of methyl 3-(N-methylacetamido)pyridine-4-carboxylate (6 g, 28.816 mmol, 1 equiv.) in dry 1,4-dioxane (100 mL) was added t-BuOK (6.47 g, 57.632 mmol, 2 equiv.) at room temperature under a nitrogen atmosphere. The resulting mixture was stirred at 90° C. under a nitrogen atmosphere for 1 hour. The reaction was monitored by LCMS. The mixture was allowed to cool to room temperature. The resulting mixture was concentrated in vacuo. The residue was purified by silica gel column chromatography eluting with DCM / MeOH (10:1) to give 4-hydroxy-1-methyl-1,2-dihydro-1,7-naphthyridin-2-one (4.5 g, 88.64%) as an orange solid.

[0362] 4-Chloro-1-methyl-1,2-dihydro-1,7-naphthyridin-2-one To a stirred solution of 4-hydroxy-1-methyl-1,2-dihydro-1,7-naphthyridin-2-one (4.5 g, 25.543 mmol, 1 equiv.) in dry 1,4-dioxane (100 mL) was added POCl (3.92 g, 25.543 mmol, 1 equiv.). The resulting mixture was stirred at 90 °C for 16 h. The mixture was allowed to cool to room temperature. The resulting mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with DCM / MeOH (10:1) to give 4-chloro-1-methyl-1,2-dihydro-1,7-naphthyridin-2-one (2 g, 40.23%) as a red solid.

[0363] 4-[[4-Fluoro-2-(trifluoromethyl)phenyl]amino]-1-methyl-1,2-dihydro-1,7-naphthyridin-2-one To a stirred solution of 4-chloro-1-methyl-1,2-dihydro-1,7-naphthyridin-2-one (0.8 g, 4.111 mmol, 1 equiv.) in dry 1,4-dioxane (15 mL), CsCO (2.68 g, 8.221 mmol, 2 equiv.), 4-fluoro-2-(trifluoromethyl)aniline (1.47 g, 8.221 mmol, 2.00 equiv.), Xantphos (0.95 g, 1.644 mmol, 0.4 equiv.), and Pd(AcO) (0.18 g, 0.822 mmol, 0.2 equiv.) were added under a nitrogen atmosphere at room temperature. The final reaction mixture was irradiated by microwave irradiation at 110 °C for 4 h. The reaction was monitored by LCMS. The resulting mixture was extracted with EtOAc (3 × 50 mL). The combined organic layers were washed with brine (1 × 100 mL) and dried over anhydrous NaSO. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography using the following conditions: (Column = spherical C18 column, 330 g, Mobile phase A = water (10 mM AcOH), Mobile phase B = acetonitrile, Flow rate = 50 mL / min, Gradient = 20% B to 40% B in 40 min, 254 / 220 nm) to give 4-[[4-fluoro-2-(trifluoromethyl)phenyl]amino]-1-methyl-1,2-dihydro-1,7-naphthyridin-2-one (1.1 g, 79.34%) as an off-white solid.

[0364] 4-[[4-Fluoro-2-(trifluoromethyl)phenyl]amino]-1-methyl-1,2,5,6,7,8-hexahydro-1,7-naphthyridin-2-one To a stirred solution of 4-[[4-fluoro-2-(trifluoromethyl)phenyl]amino]-1-methyl-1,2-dihydro-1,7-naphthyridin-2-one (1 g, 2.965 mmol, 1 equiv.) in THF (20 mL) was added PtO2 (67.33 mg, 0.296 mmol, 0.10 equiv.) at room temperature under a nitrogen atmosphere. The resulting mixture was stirred at room temperature under a hydrogen atmosphere for 16 hours. The reaction was monitored by LCMS. The resulting mixture was filtered, and the filter cake was washed with EtOAc (3 x 20 mL). The filtrate was concentrated under reduced pressure. The resulting mixture was concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography using the following conditions: (Column: C18 column, 330 g, Mobile phase A: water (10 mM AcOH), Mobile phase B: acetonitrile, Flow rate: 80 mL / min, Gradient: 5% B to 20% B in 40 min, 254 / 220 nm). Fractions containing the desired product were collected at 16% and concentrated under reduced pressure to give 4-[[4-fluoro-2-(trifluoromethyl)phenyl]amino]-1-methyl-1,2,5,6,7,8-hexahydro-1,7-naphthyridin-2-one (750 mg, 74.11%) as an off-white solid.

[0365] 7-[5-chloro-1-(oxan-2-yl)-6-oxo-1,6-dihydropyridazin-4-yl]-4-[[4-fluoro-2-(trifluoromethyl)phenyl]amino]-1-methyl-1,2,5,6,7,8-hexahydro-1,7-naphthyridin-2-one To a stirred mixture of 4-[[4-fluoro-2-(trifluoromethyl)phenyl]amino]-1-methyl-1,2,5,6,7,8-hexahydro-1,7-naphthyridin-2-one (750 mg, 2.197 mmol, 1 equiv.) and 4,5-dichloro-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (1.09 g, 4.395 mmol, 2 equiv.), DIPEA (568.00 mg, 4.395 mmol, 2 equiv.) was added at room temperature. The resulting mixture was stirred at 100° C. for 2 hours. The reaction was monitored by LCMS. The residue was then dissolved in DMF (10 mL). The solution was purified by reverse-phase flash chromatography using the following conditions: (Column: C18 column, 330 g, Mobile phase A: water (10 mM FA), Mobile phase B: acetonitrile, Flow rate: 80 mL / min, Gradient: 30% B to 50% B in 40 min, 254 / 220 nm). Fractions containing the desired product were collected at 44% B and concentrated under reduced pressure to give 7-[5-chloro-1-(oxan-2-yl)-6-oxo-1,6-dihydropyridazin-4-yl]-4-[[4-fluoro-2-(trifluoromethyl)phenyl]amino]-1-methyl-1,2,5,6,7,8-hexahydro-1,7-naphthyridin-2-one (1 g, 82.15%) as a yellow oil.

[0366] 7-[5-chloro-1-(oxan-2-yl)-6-oxo-1,6-dihydropyridazin-4-yl]-4-[[4-fluoro-2-(trifluoromethyl)phenyl](methyl)amino]-1-methyl-1,2,5,6,7,8-hexahydro-1,7-naphthyridin-2-one To a stirred solution of 7-[5-chloro-1-(oxan-2-yl)-6-oxo-1,6-dihydropyridazin-4-yl]-4-[[4-fluoro-2-(trifluoromethyl)phenyl]amino]-1-methyl-1,2,5,6,7,8-hexahydro-1,7-naphthyridin-2-one (800 mg, 1.444 mmol, 1 equiv.) in DMF (20 mL) was added CsCO (0.94 g, 2.888 mmol, 2 equiv.) and MeI (614.96 mg, 4.333 mmol, 3 equiv.) at room temperature. The resulting mixture was stirred at room temperature for 16 hours. The reaction was monitored by LCMS. The mixture was purified by reverse-phase flash chromatography using the following conditions: column = C18 column, 120 g, mobile phase A = water (10 mM AcOH), mobile phase B = acetonitrile, flow rate = 60 mL / min, gradient = 40% B to 60% B in 40 min, 254 / 220 nm. Fractions containing the desired product were collected at 49% B and concentrated under reduced pressure to give 7-[5-chloro-1-(oxan-2-yl)-6-oxo-1,6-dihydropyridazin-4-yl]-4-[[4-fluoro-2-(trifluoromethyl)phenyl](methyl)amino]-1-methyl-1,2,5,6,7,8-hexahydro-1,7-naphthyridin-2-one (80 mg, 9.75%) as a yellow oil.

[0367] 7-(5-chloro-6-oxo-1,6-dihydropyridazin-4-yl)-4-[[4-fluoro-2-(trifluoromethyl)phenyl](methyl)amino]-1-methyl-1,2,5,6,7,8-hexahydro-1,7-naphthyridin-2-one To a stirred solution of 7-[5-chloro-1-(oxan-2-yl)-6-oxo-1,6-dihydropyridazin-4-yl]-4-[[4-fluoro-2-(trifluoromethyl)phenyl](methyl)amino]-1-methyl-1,2,5,6,7,8-hexahydro-1,7-naphthyridin-2-one (80 mg, 0.141 mmol, 1 equiv.) in DCM (4.5 mL) was added TFA (0.5 mL, 6.732 mmol, 31.07 equiv.) dropwise at room temperature. The resulting mixture was stirred at room temperature for 2 hours. The reaction was monitored by LCMS. The resulting mixture was concentrated under reduced pressure. The residue was basified to pH 8 with saturated NaHCO3 (aq.). The solution was purified by reverse-phase flash to give 7-(5-chloro-6-oxo-1,6-dihydropyridazin-4-yl)-4-[[4-fluoro-2-(trifluoromethyl)phenyl](methyl)amino]-1-methyl-1,2,5,6,7,8-hexahydro-1,7-naphthyridin-2-one (40 mg, 58.69%) as a white solid.

[0368] Example 21 Synthesis of Compounds 135 and 137

[0369] [ka] tert-Butyl 2-chloro-4-(4-fluoro-2-(trifluoromethyl)phenoxy)-5,8-dihydropyrido[3,4-d]pyrimidine-7(6H)-carboxylate To a solution of tert-butyl 2,4-dichloro-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (5 g, 16.44 mmol) in DMF (50 mL) were added 4-fluoro-2-(trifluoromethyl)phenol (4.44 g, 24.66 mmol) and KCO (3.41 g, 24.66 mmol) at room temperature. The resulting mixture was stirred at 70 °C for 1 hour. After cooling to room temperature, it was filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography using the following conditions: (Column = spherical C18, 20-40 μm, 330 g, Mobile phase A = water (+ 5 mM NH4HCO3), Mobile phase B = acetonitrile, Flow rate = 80 mL / min, Gradient = 5% B in 10 min, Gradient = 35% B to 45% B in 10 min, Detector = 254 nm / 220 nm). Fractions containing the desired product were collected at 44% B and concentrated under reduced pressure to give tert-butyl 2-chloro-4-[4-fluoro-2-(trifluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (6.2 g, 85%) as a white solid.

[0370] tert-Butyl 4-(4-fluoro-2-(trifluoromethyl)phenoxy)-2-vinyl-5,8-dihydropyrido[3,4-d]pyrimidine-7(6H)-carboxylate To a solution of tert-butyl 2-chloro-4-[4-fluoro-2-(trifluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (500 mg, 1.12 mmol) in dioxane (10 mL) was added 2-ethenyl-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (344 mg, 2.23 mmol), HO (0.5 mL, 27.75 mmol), KCO (309 mg, 2.23 mmol), and Pd(PPh) (129 mg, 0.11 mmol). After stirring at 95 °C under a nitrogen atmosphere for 2 h, the resulting mixture was concentrated under reduced pressure. The residue was purified by preparative TLC, eluting with 17% ethyl acetate in petroleum ether to give tert-butyl 2-ethenyl-4-[4-fluoro-2-(trifluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (490 mg, 99%) as a pale yellow solid.

[0371] tert-Butyl 2-(1,2-dihydroxyethyl)-4-(4-fluoro-2-(trifluoromethyl)phenoxy)-5,8-dihydropyrido[3,4-d]pyrimidine-7(6H)-carboxylate To a solution of tert-butyl 2-ethenyl-4-[4-fluoro-2-(trifluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (400 mg, 0.91 mmol) in DCM (20 mL) was added 4-hydroxy-4-methylmorpholin-4-ium (323 mg, 2.73 mmol) and KOsO.2HO (34 mg, 0.091 mmol) at room temperature. After stirring for an additional 1 h, the resulting mixture was concentrated under reduced pressure, and the residue was purified by reverse-phase flash chromatography using the following conditions: column = spherical C18, 20-40 μm, 120 g, mobile phase A = water (+ 5 mM NH4HCO3), mobile phase B = acetonitrile, flow rate = 45 mL / min, gradient = 5% B in 10 min, 45% B to 65% B in 15 min, detector = 254 nm and 220 nm. Fractions containing the desired product were collected at 64% B and concentrated under reduced pressure to give tert-butyl 2-(1,2-dihydroxyethyl)-4-[4-fluoro-2-(trifluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (280 mg, 65%) as a white solid.

[0372] 1-(4-(4-fluoro-2-(trifluoromethyl)phenoxy)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)ethane-1,2-diol To a stirred solution of tert-butyl 2-(1,2-dihydroxyethyl)-4-[4-fluoro-2-(trifluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (280 mg, 0.59 mmol) in DCM (4 mL) was added TFA (1 mL) at room temperature. The resulting mixture was stirred at room temperature for 1 hour. The resulting mixture was concentrated in vacuo. The residue was dissolved in DCM (50 mL) and washed with saturated aqueous NaHCO3 (20 mL). The organic layer was separated and dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure, and the residue was purified by preparative TLC using 8% methanol in dichloromethane to give 1-[4-[4-fluoro-2-(trifluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-2-yl]ethane-1,2-diol (180 mg, 82%) as a brown solid.

[0373] 4-chloro-5-(2-(1,2-dihydroxyethyl)-4-(4-fluoro-2-(trifluoromethyl)phenoxy)-5,8-dihydropyrido[3,4-d]pyrimidin-7(6H)-yl)-2-(tetrahydro-2H-pyran-2-yl)pyridazin-3(2H)-one To a stirred solution of 2-[5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-4-yloxy]benzaldehyde (180 mg, 0.71 mmol) in DIEA (0.5 mL) was added 4,5-dichloro-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (176 mg, 0.71 mmol) at room temperature. The resulting mixture was stirred at 90° C. for 1 hour. After cooling to ambient temperature, the resulting mixture was concentrated under reduced pressure. The residue was purified by preparative TLC, eluting with 8% methanol in dichloromethane to give 4-chloro-5-(2-(1,2-dihydroxyethyl)-4-(4-fluoro-2-(trifluoromethyl)phenoxy)-5,8-dihydropyrido[3,4-d]pyrimidin-7(6H)-yl)-2-(tetrahydro-2H-pyran-2-yl)pyridazin-3(2H)-one (140 mg, 43%) as a brown solid.

[0374] (S)-4-chloro-5-(2-(1,2-dihydroxyethyl)-4-(4-fluoro-2-(trifluoromethyl)phenoxy)-5,8-dihydropyrido[3,4-d]pyrimidin-7(6H)-yl)pyridazin-3(2H)-one and (R)-4-chloro-5-(2-(1,2-dihydroxyethyl)-4-(4-fluoro-2-(trifluoromethyl)phenoxy)-5,8-dihydropyrido[3,4-d]pyrimidin-7(6H)-yl)pyridazin-3(2H)-one To a solution of 4-chloro-5-[2-(1,2-dihydroxyethyl)-4-[4-fluoro-2-(trifluoromethyl)phenoxy]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl]-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (150 mg, 0.27 mmol) in DCM (4 mL) was added TFA (1 mL) at room temperature. The resulting mixture was stirred at room temperature for 1 hour. The resulting mixture was concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography using the following conditions: column = spherical C18, 20-40 μm, 120 g, mobile phase A = water (+ 5 mM NH4HCO3), mobile phase B = acetonitrile, flow rate = 45 mL / min, gradient = 5% B in 10 min, 45% B to 65% B in 15 min, detector = 254 nm and 220 nm. Fractions containing the desired product were collected at 64% B and concentrated under reduced pressure to give the racemic product (130 mg), which was separated by preparative chiral HPLC using the following conditions: column = XBridge Prep OBD C18 column, 30 × 150 mm, 5 μm, mobile phase A = hexane, mobile phase B = EtOH, flow rate = 20 mL / min, gradient = 35% B in 10 min, detector = 254 / 220 nm. Two isomers were separated by this technique, but the absolute orientation was not determined. Fractions containing the desired product were collected and concentrated under reduced pressure to give the following products: (S)-4-chloro-5-(2-(1,2-dihydroxyethyl)-4-(4-fluoro-2-(trifluoromethyl)phenoxy)-5,8-dihydropyrido[3,4-d]pyrimidin-7(6H)-yl)pyridazin-3(2H)-one (retention time (4.97 min) (49.5 mg, 39%)) as a white solid, and (R)-4-chloro-5-(2-(1,2-dihydroxyethyl)-4-(4-fluoro-2-(trifluoromethyl)phenoxy)-5,8-dihydropyrido[3,4-d]pyrimidin-7(6H)-yl)pyridazin-3(2H)-one (retention time (8.05 min) (45.7 mg, 36%)) as a white solid.

[0375] Example 22 Synthesis of Compound 131

[0376] [ka] tert-Butyl 4-[[4-(trifluoromethyl)pyridin-3-yl]amino]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate To a stirred mixture of tert-butyl 4-chloro-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (500 mg, 1.854 mmol, 1 equiv.) and 4-(trifluoromethyl)pyridin-3-amine (601.03 mg, 3.707 mmol, 2.0 equiv.) in 1,4-dioxane (5 mL), Pd(AcO) (83.24 mg, 0.371 mmol, 0.2 equiv.), CsCO (1207.95 mg, 3.707 mmol, 2.0 equiv.), and Xantphos (429.04 mg, 0.741 mmol, 0.4 equiv.) were added at room temperature under a nitrogen atmosphere. The resulting mixture was stirred at 110 °C under a nitrogen atmosphere for 2 hours. The reaction was monitored by LCMS. The mixture was allowed to cool to room temperature. The resulting mixture was filtered, and the filter cake was washed with DCM (3 × 2 mL). The filtrate was concentrated under reduced pressure. The resulting mixture was concentrated under reduced pressure. The crude product was purified by reverse-phase flash chromatography using the following conditions: (Column: C18, 120 g, Mobile phase A: water / 0.05% NH4HCO3, Mobile phase B: ACN, Flow rate: 45 mL / min, Gradient: 45% B to 65% B in 15 min, Detector: 254 nm and 220 nm, Desired product collected at 64% B) to give tert-butyl 4-[[4-(trifluoromethyl)pyridin-3-yl]amino]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (600 mg, 81.86%) as a white solid.

[0377] tert-Butyl 4-[methyl[4-(trifluoromethyl)pyridin-3-yl]amino]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate To a stirred mixture of tert-butyl 4-[[4-(trifluoromethyl)pyridin-3-yl]amino]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (1.32 g, 3.339 mmol, 1 equiv.) and CsCO (2.18 g, 6.677 mmol, 2.0 equiv.) in DMF (10 mL) was added CHI (0.95 g, 6.677 mmol, 2.0 equiv.) under a nitrogen atmosphere at 0° C. The resulting mixture was stirred at room temperature under a nitrogen atmosphere for 2 hours. The reaction was monitored by LCMS. The crude product was purified by reverse-phase flash using the following conditions: (Column = C18, 120 g, Mobile phase A = water / 0.05% NH4HCO3, Mobile phase B = ACN, Flow rate = 45 mL / min, Gradient = 45% B to 65% B in 15 min, Detector = 254 nm and 220 nm, desired product collected at 64% B) to give tert-butyl 4-[methyl[4-(trifluoromethyl)pyridin-3-yl]amino]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (400 mg, 29.26%) as a brown solid.

[0378] N-Methyl-N-[5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-4-yl]-4-(trifluoromethyl)pyridin-3-amine To a stirred solution of tert-butyl 4-[methyl[4-(trifluoromethyl)pyridin-3-yl]amino]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidine-7-carboxylate (220 mg, 0.537 mmol, 1 equiv.) in DCM (4 mL) was added TFA (1 mL) at room temperature under a nitrogen atmosphere. The resulting mixture was stirred at room temperature for 1 hour. The reaction was monitored by LCMS. The resulting mixture was concentrated under reduced pressure. The mixture was basified to pH 8 with saturated NaHCO3 (aq.). The resulting mixture was concentrated under reduced pressure. The residue was purified by preparative TLC (DCM / MeOH=12:1) to afford N-methyl-N-[5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-4-yl]-4-(trifluoromethyl)pyridin-3-amine (130 mg, 78.22%) as a brown solid.

[0379] 4-chloro-5-(4-[methyl[4-(trifluoromethyl)pyridin-3-yl]amino]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl)-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one To a stirred solution of N-methyl-N-[5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-4-yl]-4-(trifluoromethyl)pyridin-3-amine (130 mg, 0.420 mmol, 1 equiv.) in DIEA (0.5 mg) was added 4,5-dichloro-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (104.69 mg, 0.420 mmol, 1.0 equiv.) at room temperature. The resulting mixture was stirred at 90° C. for 1 hour. The reaction was monitored by LCMS. The mixture was allowed to cool to room temperature. The resulting mixture was concentrated under reduced pressure. The residue was purified by preparative TLC (DCM / MeOH=12:1) to give 4-chloro-5-(4-[methyl[4-(trifluoromethyl)pyridin-3-yl]amino]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl)-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (100 mg, 45.58%) as a brown solid.

[0380] 4-chloro-5-(4-[methyl[4-(trifluoromethyl)pyridin-3-yl]amino]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl)-2,3-dihydropyridazin-3-one To a stirred solution of 4-chloro-5-(4-[methyl[4-(trifluoromethyl)pyridin-3-yl]amino]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl)-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (100 mg, 0.192 mmol, 1 equiv) in DCM (4 mL) was added TFA (1 mL) at room temperature. The resulting mixture was stirred at room temperature for 1 hour. The reaction was monitored by LCMS. The resulting mixture was concentrated under reduced pressure. The mixture was basified to pH 8 with saturated NaHCO3 (aq). The resulting mixture was concentrated under reduced pressure. The crude product (100 mg) was purified by preparative HPLC using the following conditions: (Column = XBridge Prep Phenyl OBD Column, 19 × 150 mm, 5 um, 13 nm, Mobile phase A = water, 5 mM NH4HCO3, Mobile phase B = acetonitrile, Flow rate = 60 mL / min, Gradient = 35% B to 55% B in 8 min, 220 nm, Rt = 7.13 min) to give 4-chloro-5-(4-[methyl[4-(trifluoromethyl)pyridin-3-yl]amino]-5H,6H,7H,8H-pyrido[3,4-d]pyrimidin-7-yl)-2,3-dihydropyridazin-3-one (52 mg, 61.99%) as a white solid.

[0381] Example 23 Synthesis of intermediates A. 2-(Difluoromethyl)-4-fluorophenyl acetate

[0382] [ka] 4-Fluoro-2-formylphenyl acetate To a solution of 5-fluoro-2-hydroxybenzaldehyde (10 g, 71.371 mmol, 1 equiv) in pyridine (100 mL, 1242.353 mmol, 17.41 equiv) was added acetyl acetate (14.57 g, 0.143 mmol, 2 equiv) at 25° C. The solution was stirred at 25° C. for 30 minutes. The resulting solution was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with PE / EA (100 / 1 to 20 / 1) to give 4-fluoro-2-formylphenyl acetate (12 g, 92.31%) as a pale yellow oil.

[0383] 2-(Difluoromethyl)-4-fluorophenyl acetate To a solution of 4-fluoro-2-formylphenyl acetate (12 g, 65.880 mmol, 1 equiv.) in DCM (200 mL, 3146.009 mmol, 47.75 equiv.) was added DAST (21.24 g, 131.760 mmol, 2 equiv.) at 0 °C. The solution was stirred at 25 °C for 4 h. The resulting solution was quenched with water (100 mL). The resulting mixture was extracted with DCM (100 mL × 2). The combined organic layers were washed with saturated aqueous NaCl solution (100 mL × 2) and dried over anhydrous NaSO. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with PE / EA (10 / 1 to 5 / 1) to give 2-(difluoromethyl)-4-fluorophenyl acetate (10 g, 74.35%) as a pale yellow oil.

[0384] B. 2-(Difluoromethyl)-4-fluorophenol

[0385] [ka] 1-Bromo-2-(difluoromethyl)-4-fluorobenzene To a stirred solution of 2-bromo-5-fluorobenzaldehyde (10 g, 49.26 mmol, 1 equiv.) in DCM (60 mL) was added DAST (15.9 g, 98.52 mmol, 2 equiv.). The resulting mixture was stirred at −10° C. for 2 h. The reaction was quenched with water at −10° C. The resulting mixture was extracted with EtOAc (4×30 mL). The combined organic layers were washed with brine (2×40 mL) and dried over anhydrous NaSO. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with PE / EtOAc (6:1) to afford 1-bromo-2-(difluoromethyl)-4-fluorobenzene (8 g, 72.18%) as a pale yellow oil.

[0386] 2-[2-(difluoromethyl)-4-fluorophenyl]-4,4,5,5-tetramethyl-1,3,2-dioxaborolane To a solution of 1-bromo-2-(difluoromethyl)-4-fluorobenzene (31 g, 137.773 mmol, 1 equiv.) and BPD (52.48 g, 206.664 mmol, 1.50 equiv.) in 1,4-dioxane (300 mL, 3541.225 mmol, 25.70 equiv.), AcOK (27.04 g, 275.546 mmol, 2 equiv.) and Pd(dppf)Cl.CHCl (5.63 g, 6.889 mmol, 0.05 equiv.) were added under a nitrogen atmosphere at 25 °C. The mixture was stirred at 90 °C for 2 h. The resulting mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with PE / EA (10 / 1) to give 2-[2-(difluoromethyl)-4-fluorophenyl]-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (30 g, 80.03%) as a pale yellow oil. The reaction was monitored by TLC. The crude product was used directly in the next step.

[0387] 2-(Difluoromethyl)-4-fluorophenol To a solution of 2-[2-(difluoromethyl)-4-fluorophenyl]-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (50 g, 183.776 mmol, 1 equiv) in MeOH (300 mL, 7409.673 mmol, 40.32 equiv) and HO (100 mL, 5550.837 mmol, 30.20 equiv), HO (30%) (50 mL, 2146.131 mmol, 11.68 equiv) was added dropwise at 0 °C. The solution was stirred at 25 °C for 3 h. The resulting solution was concentrated under reduced pressure. The residue was diluted with EA (500 mL). The organic layer was washed 3 × 200 mL with saturated NaCl (aq). The combined organic layers were dried over anhydrous Na2SO4 and concentrated under reduced pressure to give 2-(difluoromethyl)-4-fluorophenol (25 g, 83.91%) as a pale yellow oil.

[0388] Example 24 Synthesis of Compound MS

[0389] [ka] tert-Butyl 1-[1-(2-bromopyridin-3-yl)ethyl]-1H,4H,5H,6H,7H-[1,2,3]triazolo[4,5-c]pyridine-5-carboxylate To a stirred mixture of 1-(2-bromopyridin-3-yl)ethan-1-amine (1009.1 mg, 5.02 mmol, 2.00 equiv.) and tert-butyl 4-oxopiperidine-1-carboxylate (500 mg, 2.51 mmol, 1 equiv.) in DMF (10 mL), 1-azido-4-nitrobenzene (576.6 mg, 3.51 mmol, 1.40 equiv.) and Zn(OAc) (460.5 mg, 2.51 mmol, 1.00 equiv.) were added in small portions at room temperature under a nitrogen atmosphere. The resulting mixture was stirred at 60° C. for 16 hours under a nitrogen atmosphere. The reaction was monitored by LCMS. The mixture was allowed to cool to room temperature. The residue was purified by reverse-phase flash using the following conditions (Column = XBridge Shield RP18OBD column, 20-40 μm, 19*150 mm, Mobile phase A = water (10 mmol / L NH4HCO3), Mobile phase B = ACN, Flow rate = 80 mL / min, Gradient = 40% B to 80% B in 30 min, 220 nm, Rt = 7.08 min) to give tert-butyl 1-[1-(2-bromopyridin-3-yl)ethyl]-1H,4H,5H,6H,7H-[1,2,3]triazolo[4,5-c]pyridine-5-carboxylate (800 mg, 78.08%) as a yellow oil.

[0390] tert-Butyl 1-[1-(2-ethenylpyridin-3-yl)ethyl]-1H,4H,5H,6H,7H-[1,2,3]triazolo[4,5-c]pyridine-5-carboxylate To a stirred mixture of tert-butyl 1-[1-(2-bromopyridin-3-yl)ethyl]-1H,4H,5H,6H,7H-[1,2,3]triazolo[4,5-c]pyridine-5-carboxylate (800 mg, 1.96 mmol, 1 equiv.) and 2-ethenyl-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (301.8 mg, 1.96 mmol, 1.00 equiv.) in dioxane (30 mL) and HO (6 mL), Pd(PPh) (226.4 mg, 0.20 mmol, 0.10 equiv.) and KCO (812.4 mg, 5.88 mmol, 3.00 equiv.) were added in small portions at room temperature under a nitrogen atmosphere. The resulting mixture was stirred at 90 °C for 16 h under a nitrogen atmosphere. The reaction was monitored by LCMS. The mixture was allowed to cool to room temperature. The resulting mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with PE / EtOAc (30 / 1 to 5 / 1) to give tert-butyl 1-[1-(2-ethenylpyridin-3-yl)ethyl]-1H,4H,5H,6H,7H-[1,2,3]triazolo[4,5-c]pyridine-5-carboxylate (600 mg, 86.15%) as a yellow oil.

[0391] tert-Butyl 1-[1-(2-ethylpyridin-3-yl)ethyl]-1H,4H,5H,6H,7H-[1,2,3]triazolo[4,5-c]pyridine-5-carboxylate To a solution of 1-[1-(2-ethenylpyridin-3-yl)ethyl]-1H,4H,5H,6H,7H-[1,2,3]triazolo[4,5-c]pyridine-5-carboxylate (300 mg, 0.84 mmol, 1 equiv.) in 20 mL of MeOH was added Pd / C (10%, 0.02 g) in a 50 mL round-bottom flask under a nitrogen atmosphere. The mixture was hydrogenated under a hydrogen atmosphere using a hydrogen balloon at room temperature for 2 hours, filtered through a Celite pad, and concentrated under reduced pressure to afford tert-butyl 1-[1-(2-ethylpyridin-3-yl)ethyl]-1H,4H,5H,6H,7H-[1,2,3]triazolo[4,5-c]pyridine-5-carboxylate (260 mg, 86.18%) as a yellow oil.

[0392] 2-Ethyl-3-(1-[1H,4H,5H,6H,7H-[1,2,3]triazolo[4,5-c]pyridin-1-yl]ethyl)pyridine To a stirred solution of tert-butyl 1-[1-(2-ethylpyridin-3-yl)ethyl]-1H,4H,5H,6H,7H-[1,2,3]triazolo[4,5-c]pyridine-5-carboxylate (260 mg, 0.73 mmol, 1 equiv.) in DCM (10 mL) was added TFA (1 mL, 13.46 mmol, 18.51 equiv.) dropwise at room temperature. The reaction mixture was stirred at room temperature for 16 hours. The reaction was monitored by LCMS. The resulting mixture was concentrated under reduced pressure. The residue was basified to pH = 8 with saturated NH4HCO3 (aq.). The resulting mixture was extracted with CHCl2 (3 x 100 mL). The combined organic layers were washed with brine (1 x 100 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC (PE / EtOAc=1 / 1) to give 2-ethyl-3-(1-[1H,4H,5H,6H,7H-[1,2,3]triazolo[4,5-c]pyridin-1-yl]ethyl)pyridine (150 mg, 80.14%) as a yellow oil.

[0393] 4-chloro-5-[1-[1-(2-ethylpyridin-3-yl)ethyl]-1H,4H,5H,6H,7H-[1,2,3]triazolo[4,5-c]pyridin-5-yl]-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one A 25 mL round-bottom flask was charged with 2-ethyl-3-(1-[1H,4H,5H,6H,7H-[1,2,3]triazolo[4,5-c]pyridin-1-yl]ethyl)pyridine (150 mg, 0.58 mmol, 1 equiv.), 4,5-dichloro-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (290.4 mg, 1.17 mmol, 2.00 equiv.), and DIEA (150.7 mg, 1.17 mmol, 2.00 equiv.) under a nitrogen atmosphere at room temperature. The resulting mixture was stirred at 90° C. for 16 hours under a nitrogen atmosphere. The residue was purified by preparative TLC (PE / EtOAc=1 / 1) to give 4-chloro-5-[1-[1-(2-ethylpyridin-3-yl)ethyl]-1H,4H,5H,6H,7H-[1,2,3]triazolo[4,5-c]pyridin-5-yl]-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (145 mg, 52.93%) as a yellow oil.

[0394] 4-chloro-5-[1-[(1R)-1-(2-ethylpyridin-3-yl)ethyl]-1H,4H,5H,6H,7H-[1,2,3]triazolo[4,5-c]pyridin-5-yl]-2,3-dihydropyridazin-3-one and 4-chloro-5-[1-[(1S)-1-(2-ethylpyridin-3-yl)ethyl]-1H,4H,5H,6H,7H-[1,2,3]triazolo[4,5-c]pyridin-5-yl]-2,3-dihydropyridazin-3-one To a stirred solution of 4-chloro-5-[1-[1-(2-ethylpyridin-3-yl)ethyl]-1H,4H,5H,6H,7H-[1,2,3]triazolo[4,5-c]pyridin-5-yl]-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (145 mg, 0.31 mmol, 1 equiv.) in DCM (10 mL) was added TFA (1 mL, 13.46 mmol, 43.64 equiv.) dropwise at room temperature. The reaction mixture was stirred at room temperature for 4 hours. The reaction was monitored by LCMS. The resulting mixture was concentrated under reduced pressure. The residue was basified to pH = 8 with saturated NH4HCO3 (aq.). The resulting mixture was extracted with CHCl2 (3 x 100 mL). The combined organic layers were washed with brine (1 x 100 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure, and the residue (75 mg) was purified by chiral preparative HPLC using the following conditions: (Column: CHIRALPAK IE, 2 × 25 cm, 5 μm, Mobile phase: (Hex / DCM = 3 / 1) / EtOH = 80 / 20, Flow rate: 20 mL / min, Gradient: 20B to 20B in 20 min, 220 / 254 nm, RT1 = 12.678, RT2 = 16.738). 4-Chloro-5-[1-[(1R)-1-(2-ethylpyridin-3-yl)ethyl]-1H,4H,5H,6H,7H-[1,2,3]triazolo[4,5-c]pyridin-5-yl]-2,3-dihydropyridazin-3-one (16.8 mg, 14.11%) was obtained as a white solid in 1.380 minutes, and 4-chloro-5-[1-[(1S)-1-(2-ethylpyridin-3-yl)ethyl]-1H,4H,5H,6H,7H-[1,2,3]triazolo[4,5-c]pyridin-5-yl]-2,3-dihydropyridazin-3-one (19.8 mg) was obtained as a white solid in 1.832 minutes (E01224-021).

[0395] Example 25 Synthesis of MX

[0396] [ka] tert-Butyl (4R)-4-methyl-1-[1-[2-(trifluoromethyl)phenyl]ethyl]-1H,4H,5H,6H,7H-[1,2,3]triazolo[4,5-c]pyridine-5-carboxylate To a stirred solution of tert-butyl (2R)-2-methyl-4-oxopiperidine-1-carboxylate (1 g, 4.69 mmol, 1 equiv.) and 1-[2-(trifluoromethyl)phenyl]ethan-1-amine (0.9 g, 4.76 mmol, 1.01 equiv.) in DMF (20 mL) was added 1-azido-4-nitrobenzene (1.1 g, 6.56 mmol, 1.4 equiv.) and Zn(OAc) (0.9 g, 4.69 mmol, 1 equiv.). The resulting mixture was stirred at 60 °C overnight, and the residue was purified by reverse-phase flash chromatography using the following conditions: column: C18 silica gel, mobile phase: MeCN in water, gradient: 20% to 60% in 40 min, detector: UV 254 nm. This gave tert-butyl (4R)-4-methyl-1-[1-[2-(trifluoromethyl)phenyl]ethyl]-1H,4H,5H,6H,7H-[1,2,3]triazolo[4,5-c]pyridine-5-carboxylate (1.5 g, 77.94%) as an off-white solid.

[0397] (4R)-4-Methyl-1-[1-[2-(trifluoromethyl)phenyl]ethyl]-1H,4H,5H,6H,7H-[1,2,3]triazolo[4,5-c]pyridine To a stirred solution of tert-butyl (4R)-4-methyl-1-[1-[2-(trifluoromethyl)phenyl]ethyl]-1H,4H,5H,6H,7H-[1,2,3]triazolo[4,5-c]pyridine-5-carboxylate (1.5 g, 3.65 mmol, 1 equiv.) in DCM (9 mL) was added TFA (3 mL), and the resulting mixture was stirred at room temperature for 2 h. The mixture was basified to pH 8 with saturated NH4HCO3 (aq). The solution was concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography using the following conditions: column = C18 silica gel, mobile phase = MeCN in water, 10% to 50% gradient in 30 min, detector = UV 254 nm. This gave (4R)-4-methyl-1-[1-[2-(trifluoromethyl)phenyl]ethyl]-1H,4H,5H,6H,7H-[1,2,3]triazolo[4,5-c]pyridine (1 g, 88.17%) as a yellow solid.

[0398] 4-chloro-5-[(6R)-6-methyl-1-[1-[2-(trifluoromethyl)phenyl]ethyl]-1H,4H,5H,6H,7H-[1,2,3]triazolo[4,5-c]pyridin-5-yl]-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one To a stirred solution of (6R)-6-methyl-1-[1-[2-(trifluoromethyl)phenyl]ethyl]-1H,4H,5H,6H,7H-[1,2,3]triazolo[4,5-c]pyridine (200 mg, 0.64 mmol, 1 equiv.), 4,5-dichloro-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (192.6 mg, 0.77 mmol, 1.2 equiv.), and DIEA (249.9 mg, 1.93 mmol, 3 equiv.) was added. The resulting mixture was stirred at 100 °C overnight. The residue was purified by reverse-phase flash chromatography using the following conditions: column = C18 silica gel, mobile phase = MeCN in water, gradient from 20% to 60% in 40 min, detector = UV 254 nm. This gave 4-chloro-5-[(6R)-6-methyl-1-[1-[2-(trifluoromethyl)phenyl]ethyl]-1H,4H,5H,6H,7H-[1,2,3]triazolo[4,5-c]pyridin-5-yl]-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (250 mg, 74.17%) as a yellow solid.

[0399] 4-chloro-5-[(6R)-6-methyl-1-[(1R)-1-[2-(trifluoromethyl)phenyl]ethyl]-1H,4H,5H,6H,7H-[1,2,3]triazolo[4,5-c]pyridin-5-yl]-2,3-dihydropyridazin-3-one and 4-chloro-5-[(6R)-6-methyl-1-[(1S)-1-[2-(trifluoromethyl)phenyl]ethyl]-1H,4H,5H,6H,7H-[1,2,3]triazolo[4,5-c]pyridin-5-yl]-2,3-dihydropyridazin-3-one To a stirred solution of 4-chloro-5-[(6R)-6-methyl-1-[1-[2-(trifluoromethyl)phenyl]ethyl]-1H,4H,5H,6H,7H-[1,2,3]triazolo[4,5-c]pyridin-5-yl]-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (240 mg, 0.46 mmol, 1 equiv.) in DCM (6 mL) was added TFA (2 mL). The resulting mixture was stirred at room temperature for 2 hours. The resulting mixture was concentrated under reduced pressure. The crude product was purified under the following conditions: (Column: XBridge Prep OBD C18 column 30 x 150 mm 5 um; Mobile phase A: water (10 mmol / L NH4HCO3), Mobile phase B: ACN; Flow rate: 60 mL / min; Gradient: 27% B to 50% B in 7 min) Purification by preparative HPLC using a HPLC column (B; 220 nm; rt: 6.38 min) afforded 4-chloro-5-[(6R)-6-methyl-1-[(1R)-1-[2-(trifluoromethyl)phenyl]ethyl]-1H,4H,5H,6H,7H-[1,2,3]triazolo[4,5-c]pyridin-5-yl]-2,3-dihydropyridazin-3-one (22.4 mg, 11.12%) as a yellow solid and 4-chloro-5-[(6R)-6-methyl-1-[(1S)-1-[2-(trifluoromethyl)phenyl]ethyl]-1H,4H,5H,6H,7H-[1,2,3]triazolo[4,5-c]pyridin-5-yl]-2,3-dihydropyridazin-3-one (58.5 mg, 29.05%) as an off-white solid.

[0400] Example 26 Synthesis of Compound LY

[0401] [ka] 3-(1-chloroethyl)-2-ethylpyridine was prepared by the method and scheme described for 3-(1-chloropropyl)-2-ethylpyridine by using the corresponding pyridine.

[0402] [ka] 3-(1-chloropropyl 1)-2-ethylpyridine To a stirred solution of 1-(2-ethylpyridin-3-yl)propan-1-ol (300 mg, 1.82 mmol, 1 equiv.) in DCM (20 mL) was added SOCl (432.0 mg, 3.63 mmol, 2.00 equiv.) dropwise at 0° C. under a nitrogen atmosphere. The resulting mixture was stirred at room temperature under a nitrogen atmosphere for 16 hours. The reaction was monitored by LCMS. The resulting mixture was concentrated in vacuo. This afforded 3-(1-chloropropyl)-2-ethylpyridine (350 mg, 104.95%) as a yellow oil.

[0403] Step 1. 4-chloro-5-[4-[1-(2-ethylpyridin-3-yl)ethyl]piperazin-1-yl]-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one To a stirred mixture of 4-chloro-2-(oxan-2-yl)-5-(piperazin-1-yl)-2,3-dihydropyridazin-3-one (100 mg, 0.33 mmol, 1 equiv.) and 3-(1-chloroethyl)-2-ethylpyridine (68.1 mg, 0.40 mmol, 1.20 equiv.) in ACN (10 mL), KCO (92.5 mg, 0.67 mmol, 2.0 equiv.) and KI (111.1 mg, 0.67 mmol, 2.00 equiv.) were added in small portions at room temperature under a nitrogen atmosphere. The resulting mixture was stirred at 70 °C for 16 h under a nitrogen atmosphere. The reaction was monitored by LCMS. The resulting mixture was filtered, and the filter cake was washed with ACN (2 × 30 mL). The filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC (CH2Cl2 / MeOH=20 / 1) to give 4-chloro-5-[4-[1-(2-ethylpyridin-3-yl)ethyl]piperazin-1-yl]-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (120 mg, 83.00%) as a yellow oil.

[0404] Step 2. LY and LZ 4-chloro-5-[4-[(1S)-1-(2-ethylpyridin-3-yl)ethyl]piperazin-1-yl]-2,3-dihydropyridazin-3-one and 4-chloro-5-[4-[(1R)-1-(2-ethylpyridin-3-yl)ethyl]piperazin-1-yl]-2,3-dihydropyridazin-3-one To a stirred solution of 4-chloro-5-[4-[1-(2-ethylpyridin-3-yl)ethyl]piperazin-1-yl]-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (120 mg, 0.28 mmol, 1 equiv.) in DCM (10 mL) was added TFA (1 mL, 13.46 mmol, 48.46 equiv.) dropwise at room temperature. The reaction mixture was stirred at room temperature for 4 hours. The reaction was monitored by LCMS. The resulting mixture was concentrated under reduced pressure. The residue was basified to pH = 8 with saturated NH4HCO3 (aq.). The resulting mixture was extracted with CHCl2 (3 x 100 mL). The combined organic layers were washed with brine (1 x 50 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue (70 mg) was purified by chiral preparative HPLC using the following conditions: (Column: CHIRALPAK IE, 2 x 25 cm, 5 μm, Mobile phase: MTBE / EtOH = 80 / 20, Flow rate: 20 mL / min, Gradient: 20B to 20B in 20 min, 220 / 254 nm, RT1 = 12.678, RT2 = 16.738). 4-Chloro-5-[4-[(1S)-1-(2-ethylpyridin-3-yl)ethyl]piperazin-1-yl]-2,3-dihydropyridazin-3-one (9.5 mg, 9.83%) was obtained in 2.544 min as a pale yellow solid, and 4-chloro-5-[4-[(1R)-1-(2-ethylpyridin-3-yl)ethyl]piperazin-1-yl]-2,3-dihydropyridazin-3-one (14.2 mg) was obtained in 2.984 min as a pale yellow solid.

[0405] Example 27 Synthesis of LW

[0406] [ka] Step 1. tert-Butyl 4-[(2-bromopyridin-3-yl)amino]piperidine-1-carboxylate To a stirred solution of 2-bromopyridin-3-amine (600 mg, 3.468 mmol, 1 equiv.) and tert-butyl 4-oxopiperidine-1-carboxylate (690.99 mg, 3.468 mmol, 1 equiv.) in DCM (20 mL) was added AcOH (208.26 mg, 3.468 mmol, 1 equiv.) dropwise / portionwise at 0 °C under a nitrogen atmosphere. The mixture was stirred at room temperature for 2 h. NaBH(OAc) (1470.00 mg, 6.936 mmol, 2.00 equiv.) was added to the mixture at 0 °C, and the mixture was stirred at room temperature overnight. The desired product could be detected by LCMS. The reaction was quenched by adding water (40 mL) at 0 °C. The aqueous layer was extracted with CHCl (2 × 30 mL). The organic layer was concentrated under reduced pressure to give tert-butyl 4-[(2-bromopyridin-3-yl)amino]piperidine-1-carboxylate (800 mg, 64.75%) as a yellow solid.

[0407] Step 2. tert-Butyl 4-[(2-ethenylpyridin-3-yl)amino]piperidine-1-carboxylate To a solution of 2-ethenyl-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (691.71 mg, 4.491 mmol, 2 equiv.) and tert-butyl 4-[(2-bromopyridin-3-yl)amino]piperidine-1-carboxylate (800 mg, 2.246 mmol, 1 equiv.) in 1,4-dioxane (10 mL) and HO (2 mL) was added KCO (931.03 mg, 6.737 mmol, 3 equiv.) and Pd(PPh) (259.48 mg, 0.225 mmol, 0.1 equiv.). After stirring overnight at 80 °C under a nitrogen atmosphere, the resulting mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with PE / EtOAc (5:1 to 3:1) to give tert-butyl 4-[(2-ethenylpyridin-3-yl)amino]piperidine-1-carboxylate (600 mg, 88.07%) as a yellow solid.

[0408] Step 3. tert-Butyl 4-[(2-ethylpyridin-3-yl)amino]piperidine-1-carboxylate To a solution of tert-butyl 4-[(2-ethenylpyridin-3-yl)amino]piperidine-1-carboxylate (600 mg, 1.978 mmol, 1 equiv.) in 30 mL of MeOH was added Pd / C (10%, 21.05 mg) in a 250 mL round-bottom flask under a nitrogen atmosphere. The mixture was hydrogenated under a hydrogen atmosphere using a hydrogen balloon at room temperature for 3 hours, filtered through a Celite pad, and concentrated under reduced pressure to give tert-butyl 4-[(2-ethylpyridin-3-yl)amino]piperidine-1-carboxylate (590 mg, 97.68%) as a yellow solid.

[0409] Step 4. 2-Ethyl-N-(piperidin-4-yl)pyridin-3-amine To a stirred solution of tert-butyl 4-[(2-ethylpyridin-3-yl)amino]piperidine-1-carboxylate (590 mg, 1 eq.) in DCM (15 mL) was added TFA (3 mL) dropwise at 0° C. under a nitrogen atmosphere. The mixture was stirred at room temperature for 1 hour. The desired product could be detected by LCMS. The resulting mixture was concentrated under reduced pressure to give 2-ethyl-N-(piperidin-4-yl)pyridin-3-amine (390 mg, 98.34%) as a white solid.

[0410] Step 5. Compound LX 4-chloro-5-[4-[(2-ethylpyridin-3-yl)amino]piperidin-1-yl]-2,3-dihydropyridazin-3-one To a stirred solution of 2-ethyl-N-(piperidin-4-yl)pyridin-3-amine (100 mg, 0.487 mmol, 1 equiv.) and 4,5-dichloro-2,3-dihydropyridazin-3-one (80.35 mg, 0.487 mmol, 1.00 equiv.) in DMA (8 mL), DIEA (125.90 mg, 0.974 mmol, 2 equiv.) was added dropwise at room temperature under a nitrogen atmosphere. The mixture was stirred at 100° C. overnight. The desired product could be detected by LCMS. The resulting mixture was concentrated under reduced pressure. The crude product (60 mg) was purified by preparative HPLC using the following conditions: (Column = XBridge Prep OBD C18 column, 30 × 150 mm, 5 μm, Mobile phase A = water (10 mmol / L NH4HCO3), Mobile phase B = ACN, Flow rate = 60 mL / min, Gradient = 18% B to 30% B in 6.5 min, 220 nm, Rt = 5.37, 8.55 min) to give 4-chloro-5-[4-[(2-ethylpyridin-3-yl)amino]piperidin-1-yl]-2,3-dihydropyridazin-3-one (20 mg) as a white solid and 5-chloro-4-[4-[(2-ethylpyridin-3-yl)amino]piperidin-1-yl]-2,3-dihydropyridazin-3-one (7 mg) as a white solid.

[0411] Step 6. tert-Butyl 4-[ethyl(2-ethylpyridin-3-yl)amino]piperidine-1-carboxylate To a stirred solution of tert-butyl 4-[(2-ethylpyridin-3-yl)amino]piperidine-1-carboxylate (150 mg, 0.491 mmol, 1 equiv.) and acetaldehyde (32.45 mg, 0.737 mmol, 1.5 equiv.) in DCM (10 mL) was added dropwise AcOH (29.49 mg, 0.491 mmol, 1 equiv.) under a nitrogen atmosphere at 0 °C. The mixture was stirred at room temperature for 2 h. NaBHCN (92.59 mg, 1.473 mmol, 3 equiv.) was added to the mixture at 0 °C, and the mixture was stirred at room temperature overnight. The desired product could be detected by LCMS. The reaction was quenched by adding water (40 mL) at 0 °C. The aqueous layer was extracted with CHCl (2 × 30 mL). The organic layer was concentrated under reduced pressure to give tert-butyl 4-[ethyl(2-ethylpyridin-3-yl)amino]piperidine-1-carboxylate (150 mg, 91.59%) as a white solid.

[0412] Step 8. N,2-Diethyl-N-(piperidin-4-yl)pyridin-3-amine To a stirred solution of tert-butyl 4-[ethyl(2-ethylpyridin-3-yl)amino]piperidine-1-carboxylate (150 mg, 1 equiv.) in DCM (10 mL) was added TFA (2 mL) dropwise at 0° C. under a nitrogen atmosphere. The mixture was stirred at room temperature for 2 hours. The desired product could be detected by LCMS. The resulting mixture was concentrated under reduced pressure to give N,2-diethyl-N-(piperidin-4-yl)pyridin-3-amine (100 mg, 95.27%) as a yellow solid.

[0413] Step 8. Compound LW 4-chloro-5-[4-[ethyl(2-ethylpyridin-3-yl)amino]piperidin-1-yl]-2,3-dihydropyridazin-3-one To a stirred solution of N,2-diethyl-N-(piperidin-4-yl)pyridin-3-amine (60 mg, 0.26 mmol, 1 equiv.) and 4,5-dichloro-2,3-dihydropyridazin-3-one (42.4 mg, 0.26 mmol, 1.00 equiv.) in DMA (5 mL, 53.78 mmol, 209.15 equiv.), DIEA (66.5 mg, 0.51 mmol, 2 equiv.) was added at room temperature under a nitrogen atmosphere. The mixture was stirred at 100° C. overnight. The desired product could be detected by LCMS. The mixture was concentrated under reduced pressure. The crude product (50 mg) was purified by preparative HPLC using the following conditions: (Column = XBridge Prep OBD C18 column, 30 × 150 mm 5 μm, Mobile phase A = water (10 mmol / L NH4HCO3), Mobile phase B = ACN, Flow rate = 60 mL / min, Gradient = 25% B to 40% B in 8 min, 220 nm, Rt = 7.58 min) to give 4-chloro-5-[4-[ethyl(2-ethylpyridin-3-yl)amino]piperidin-1-yl]-2,3-dihydropyridazin-3-one (24.3 mg) as a white solid.

[0414] Example 28 Synthesis of OM

[0415] [ka] Compound OM was prepared by the method and scheme described for compound OK by using the corresponding amine.

[0416] [ka] Preparation of OK 5-tert-Butyl-3-ethyl 2-[[2-(difluoromethyl)phenyl]methyl]-2H,4H,5H,6H,7H-pyrazolo[4,3-c]pyridine-3,5-dicarboxylate To a stirred solution of 1-(chloromethyl)-2-(difluoromethyl)benzene (800 mg, 4.530 mmol, 1 equiv.) and 5-tert-butyl 3-ethyl 1H,4H,5H,6H,7H-pyrazolo[4,3-c]pyridine-3,5-dicarboxylate (1337.96 mg, 4.530 mmol, 1.00 equiv.) in MeCN (15 mL), KI (752.04 mg, 4.530 mmol, 1 equiv.) and KCO (1252.22 mg, 9.061 mmol, 2 equiv.) were added at room temperature under a nitrogen atmosphere. The mixture was stirred at 80 °C overnight. The desired product could be detected by LCMS. The resulting mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with PE / EtOAc (5:1 to 3:1) to give 5-tert-butyl 3-ethyl 1-[[2-(difluoromethyl)phenyl]methyl]-1H,4H,5H,6H,7H-pyrazolo[4,3-c]pyridine-3,5-dicarboxylate (500 mg, 25.34%) as a yellow solid and 5-tert-butyl 3-ethyl 2-[[2-(difluoromethyl)phenyl]methyl]-2H,4H,5H,6H,7H-pyrazolo[4,3-c]pyridine-3,5-dicarboxylate (200 mg, 10.14%) as a yellow solid.

[0417] Ethyl 1-[[2-(difluoromethyl)phenyl]methyl]-1H,4H,5H,6H,7H-pyrazolo[4,3-c]pyridine-3-carboxylate To a stirred solution of 5-tert-butyl 3-ethyl 1-[[2-(difluoromethyl)phenyl]methyl]-1H,4H,5H,6H,7H-pyrazolo[4,3-c]pyridine-3,5-dicarboxylate (500 mg, 1.148 mmol, 1 equiv.) in DCM (10 mL, 157.300 mmol, 137.00 equiv.), TFA (2 mL, 26.926 mmol, 23.45 equiv.) was added dropwise at room temperature under a nitrogen atmosphere. The mixture was stirred at room temperature for 2 hours. The desired product could be detected by LCMS. The resulting mixture was concentrated under reduced pressure to give ethyl 1-[[2-(difluoromethyl)phenyl]methyl]-1H,4H,5H,6H,7H-pyrazolo[4,3-c]pyridine-3-carboxylate (380 mg, 98.69%) as a yellow solid.

[0418] Ethyl 5-[5-chloro-1-(oxan-2-yl)-6-oxo-1,6-dihydropyridazin-4-yl]-1-[[2-(difluoromethyl)phenyl]methyl]-1H,4H,5H,6H,7H-pyrazolo[4,3-c]pyridine-3-carboxylate To a stirred solution of ethyl 1-[[2-(difluoromethyl)phenyl]methyl]-1H,4H,5H,6H,7H-pyrazolo[4,3-c]pyridine-3-carboxylate (380 mg, 1.133 mmol, 1 equiv.) in DIEA (292.90 mg, 2.266 mmol, 2 equiv.), 4,5-dichloro-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (282.25 mg, 1.133 mmol, 1.00 equiv.) was added in small portions at room temperature under a nitrogen atmosphere. The mixture was stirred at 100 °C overnight. The desired product could be detected by LCMS. The residue was purified by silica gel column chromatography, eluting with PE / EtOAc (5:1 to 3:1) to give ethyl 5-[5-chloro-1-(oxan-2-yl)-6-oxo-1,6-dihydropyridazin-4-yl]-1-[[2-(difluoromethyl)phenyl]methyl]-1H,4H,5H,6H,7H-pyrazolo[4,3-c]pyridine-3-carboxylate (500 mg, 80.52%) as a yellow solid.

[0419] 5-[5-chloro-1-(oxan-2-yl)-6-oxo-1,6-dihydropyridazin-4-yl]-1-[[2-(difluoromethyl)phenyl]methyl]-1H,4H,5H,6H,7H-pyrazolo[4,3-c]pyridine-3-carboxylic acid To a stirred solution of ethyl 5-[5-chloro-1-(oxan-2-yl)-6-oxo-1,6-dihydropyridazin-4-yl]-1-[[2-(difluoromethyl)phenyl]methyl]-1H,4H,5H,6H,7H-pyrazolo[4,3-c]pyridine-3-carboxylate (460 mg, 0.839 mmol, 1 equiv.) in THF (5 mL) and HO (5 mL), LiOH (100.51 mg, 4.197 mmol, 5 equiv.) was added portionwise at room temperature under a nitrogen atmosphere. The mixture was stirred at 50 °C overnight. The desired product could be detected by LCMS. The resulting mixture was concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography using the following conditions: Column: XBridge Prep OBD C18 column, 30 x 150 mm, 5 µm; Mobile phase A: water (10 mmol / L NH4HCO3); Mobile phase B: ACN; Flow rate: 60 mL / min; Gradient: 27% B to 55% B in 8 min; 220 nm; Rt: 7.82 min to give 5-[5-chloro-1-(oxan-2-yl)-6-oxo-1,6-dihydropyridazin-4-yl]-1-[[2-(difluoromethyl)phenyl]methyl]-1H,4H,5H,6H,7H-pyrazolo[4,3-c]pyridine-3-carboxylic acid (430 mg, 98.52%) as a colorless oil.

[0420] 5-[5-chloro-1-(oxan-2-yl)-6-oxo-1,6-dihydropyridazin-4-yl]-1-[[2-(difluoromethyl)phenyl]methyl]-N,N-dimethyl-1H,4H,5H,6H,7H-pyrazolo[4,3-c]pyridine-3-carboxamide To a stirred solution of 5-[5-chloro-1-(oxan-2-yl)-6-oxo-1,6-dihydropyridazin-4-yl]-1-[[2-(difluoromethyl)phenyl]methyl]-1H,4H,5H,6H,7H-pyrazolo[4,3-c]pyridine-3-carboxylic acid (80 mg, 0.15 mmol, 1 equiv.) in DMF (5 mL), CDI (37.4 mg, 0.23 mmol, 1.5 equiv.) was added portionwise at room temperature under a nitrogen atmosphere. The mixture was stirred at 50° C. for 2 hours. Dimethylamine (13.9 mg, 0.31 mmol, 2.00 equiv.) was added to the mixture. The mixture was stirred at 50° C. overnight. The desired product could be detected by LCMS. The resulting mixture was concentrated under vacuum to afford 5-[5-chloro-1-(oxan-2-yl)-6-oxo-1,6-dihydropyridazin-4-yl]-1-[[2-(difluoromethyl)phenyl]methyl]-N,N-dimethyl-1H,4H,5H,6H,7H-pyrazolo[4,3-c]pyridine-3-carboxamide (60 mg, 71.29%) as a yellow solid.

[0421] 5-(5-chloro-6-oxo-1,6-dihydropyridazin-4-yl)-1-[[2-(difluoromethyl)phenyl]methyl]-N,N-dimethyl-1H,4H,5H,6H,7H-pyrazolo[4,3-c]pyridine-3-carboxamide To a stirred solution of 5-[5-chloro-1-(oxan-2-yl)-6-oxo-1,6-dihydropyridazin-4-yl]-1-[[2-(difluoromethyl)phenyl]methyl]-N,N-dimethyl-1H,4H,5H,6H,7H-pyrazolo[4,3-c]pyridine-3-carboxamide (50 mg, 1 equiv.) in DCM (10 mL) was added TFA (2 mL) dropwise at room temperature under a nitrogen atmosphere. The mixture was stirred at room temperature for 1 hour. The desired product could be detected by LCMS. The resulting mixture was concentrated under reduced pressure. The crude product (30 mg) was purified by preparative HPLC using the following conditions: (Column = XBridge Prep OBD C18 column, 30 × 150 mm, 5 μm, Mobile phase A = water (10 mmol / L NH4HCO3), Mobile phase B = ACN, Flow rate = 60 mL / min, Gradient = 23% B to 45% B in 7 min, 220 nm, Rt = 6.47 min) to give 5-(5-chloro-6-oxo-1,6-dihydropyridazin-4-yl)-1-[[2-(difluoromethyl)phenyl]methyl]-N,N-dimethyl-1H,4H,5H,6H,7H-pyrazolo[4,3-c]pyridine-3-carboxamide (25 mg) as a white solid.

[0422] Example 29 Synthesis of Compound OU

[0423] [ka] Preparation of OU 5-tert-Butyl-3-ethyl 2-[[2-(difluoromethyl)phenyl]methyl]-2H,4H,5H,6H,7H-pyrazolo[4,3-c]pyridine-3,5-dicarboxylate To a stirred solution of 1-(chloromethyl)-2-(difluoromethyl)benzene (800 mg, 4.530 mmol, 1 equiv.) and 5-tert-butyl 3-ethyl 1H,4H,5H,6H,7H-pyrazolo[4,3-c]pyridine-3,5-dicarboxylate (1337.96 mg, 4.530 mmol, 1.00 equiv.) in MeCN (15 mL), KI (752.04 mg, 4.530 mmol, 1 equiv.) and KCO (1252.22 mg, 9.061 mmol, 2 equiv.) were added at room temperature under a nitrogen atmosphere. The mixture was stirred at 80 °C overnight. The desired product could be detected by LCMS. The resulting mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with PE / EtOAc (5:1 to 3:1) to give 5-tert-butyl 3-ethyl 1-[[2-(difluoromethyl)phenyl]methyl]-1H,4H,5H,6H,7H-pyrazolo[4,3-c]pyridine-3,5-dicarboxylate (500 mg, 25.34%) as a yellow solid and 5-tert-butyl 3-ethyl 2-[[2-(difluoromethyl)phenyl]methyl]-2H,4H,5H,6H,7H-pyrazolo[4,3-c]pyridine-3,5-dicarboxylate (200 mg, 10.14%) as a yellow solid.

[0424] tert-Butyl 1-[[2-(difluoromethyl)phenyl]methyl]-3-(hydroxymethyl)-1H,4H,5H,6H,7H-pyrazolo[4,3-c]pyridine-5-carboxylate To a stirred solution of 5-tert-butyl 3-ethyl 1-[[2-(difluoromethyl)phenyl]methyl]-1H,4H,5H,6H,7H-pyrazolo[4,3-c]pyridine-3,5-dicarboxylate (600 mg, 1.378 mmol, 1 equiv.) in THF (10 mL, 123.430 mmol, 89.58 equiv.), LiAlH4 (62.75 mg, 1.653 mmol, 1.2 equiv.) was added portionwise at 0 °C under a nitrogen atmosphere. The mixture was stirred at room temperature for 1 h. The desired product could be detected by LCMS. The reaction was quenched by adding water (5 mL) at 0 °C. The mixture was concentrated and purified by silica gel column chromatography (PE:EA=2:1) ​​to give tert-butyl 1-[[2-(difluoromethyl)phenyl]methyl]-3-(hydroxymethyl)-1H,4H,5H,6H,7H-pyrazolo[4,3-c]pyridine-5-carboxylate (500 mg, 92.24%) as a white solid.

[0425] (1-[[2-(difluoromethyl)phenyl]methyl]-1H,4H,5H,6H,7H-pyrazolo[4,3-c]pyridin-3-yl)methanol To a stirred solution of tert-butyl 1-[[2-(difluoromethyl)phenyl]methyl]-3-(hydroxymethyl)-1H,4H,5H,6H,7H-pyrazolo[4,3-c]pyridine-5-carboxylate (500 mg, 1.271 mmol, 1 equiv.) in DCM (10 mL, 157.300 mmol, 123.78 equiv.), TFA (2 mL, 26.926 mmol, 21.19 equiv.) was added dropwise at 0° C. under a nitrogen atmosphere. The mixture was stirred at room temperature for 2 hours. The desired product could be detected by LCMS. The mixture was concentrated to give (1-[[2-(difluoromethyl)phenyl]methyl]-1H,4H,5H,6H,7H-pyrazolo[4,3-c]pyridin-3-yl)methanol (370 mg, 99.26%) as a yellow solid.

[0426] 4-chloro-5-(1-[[2-(difluoromethyl)phenyl]methyl]-3-(hydroxymethyl)-1H,4H,5H,6H,7H-pyrazolo[4,3-c]pyridin-5-yl)-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one To a stirred solution of (1-[[2-(difluoromethyl)phenyl]methyl]-1H,4H,5H,6H,7H-pyrazolo[4,3-c]pyridin-3-yl)methanol (370 mg, 1.261 mmol, 1 equiv.) and 4,5-dichloro-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (471.31 mg, 1.892 mmol, 1.5 equiv.) in DMA (1 mL), DIEA (326.06 mg, 2.523 mmol, 2 equiv.) was added dropwise at room temperature under a nitrogen atmosphere. The mixture was stirred at 100° C. overnight. The desired product could be detected by LCMS. The resulting mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with PE / EtOAc (6:1 to 3:1) to give 4-chloro-5-(1-[[2-(difluoromethyl)phenyl]methyl]-3-(hydroxymethyl)-1H,4H,5H,6H,7H-pyrazolo[4,3-c]pyridin-5-yl)-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (420 mg, 65.81%) as a white solid.

[0427] 4-chloro-5-[3-(chloromethyl)-1-[[2-(difluoromethyl)phenyl]methyl]-1H,4H,5H,6H,7H-pyrazolo[4,3-c]pyridin-5-yl]-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one To a stirred solution of 4-chloro-5-(1-[[2-(difluoromethyl)phenyl]methyl]-3-(hydroxymethyl)-1H,4H,5H,6H,7H-pyrazolo[4,3-c]pyridin-5-yl)-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (400 mg, 0.791 mmol, 1 equiv.) and TEA (160.00 mg, 1.581 mmol, 2 equiv.) in DCM (8 mL, 125.840 mmol, 159.17 equiv.), MsCl (108.68 mg, 0.949 mmol, 1.2 equiv.) was added dropwise at 0° C. under a nitrogen atmosphere. The mixture was stirred at room temperature overnight. The desired product could be detected by LCMS. The resulting mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with PE / EtOAc (5:1) to give 4-chloro-5-[3-(chloromethyl)-1-[[2-(difluoromethyl)phenyl]methyl]-1H,4H,5H,6H,7H-pyrazolo[4,3-c]pyridin-5-yl]-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (400 mg, 96.48%) as a yellow solid.

[0428] 4-chloro-5-(1-[[2-(difluoromethyl)phenyl]methyl]-3-([4-methylpiperazin-1-yl)methyl]-1H,4H,5H,6H,7H-pyrazolo[4,3-c]pyridin-5-yl)-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one To a stirred solution of 4-chloro-5-[3-(chloromethyl)-1-[[2-(difluoromethyl)phenyl]methyl]-1H,4H,5H,6H,7H-pyrazolo[4,3-c]pyridin-5-yl]-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (60 mg, 0.103 mmol, 1 equiv.) in MeCN (10 mL) was added 1-methylpiperazine (51.45 mg, 0.514 mmol, 5 equiv.) dropwise at room temperature under a nitrogen atmosphere. The mixture was stirred at 80° C. overnight. The desired product could be detected by LCMS. The resulting mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with PE / EtOAc (5:1 to 1:1) to give 4-chloro-5-(1-[[2-(difluoromethyl)phenyl]methyl]-3-[(4-methylpiperazin-1-yl)methyl]-1H,4H,5H,6H,7H-pyrazolo[4,3-c]pyridin-5-yl)-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (60 mg, 99.31%) as a white solid.

[0429] 4-chloro-5-(1-[[2-(difluoromethyl)phenyl]methyl]-3-[(4-methylpiperazin-1-yl)methyl]-1H,4H,5H,6H,7H-pyrazolo[4,3-c]pyridin-5-yl)-2,3-dihydropyridazin-3-one To a stirred solution of 4-chloro-5-(1-[[2-(difluoromethyl)phenyl]methyl]-3-[(4-methylpiperazin-1-yl)methyl]-1H,4H,5H,6H,7H-pyrazolo[4,3-c]pyridin-5-yl)-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (60 mg) in DCM (10 mL) was added dropwise TFA (2 mL) under a nitrogen atmosphere at 0° C., and the mixture was stirred at room temperature for 2 hours. The desired product could be detected by LCMS. The reaction was quenched by adding saturated NaHCO3 (aq) (5 mL) at room temperature. The resulting mixture was concentrated under reduced pressure. The crude product (mg) was purified by preparative HPLC using the following conditions: (Column = XBridge Prep OBD C18 column, 30 × 150 mm, 5 μm, Mobile phase A = water (10 mmol / L NH4HCO3), Mobile phase B = ACN, Flow rate = 60 mL / min, Gradient = 20% B to 35% B in 8 min, 220 nm, Rt = 7.25 min) to give 4-chloro-5-(1-[[2-(difluoromethyl)phenyl]methyl]-3-[(4-methylpiperazin-1-yl)methyl]-1H,4H,5H,6H,7H-pyrazolo[4,3-c]pyridin-5-yl)-2,3-dihydropyridazin-3-one (30 mg) as a white solid.

[0430] Example 30 Synthesis of Compound OP

[0431] [ka] Preparation of OPs 2-tert-butyl 7-ethyl-5-[[2-(trifluoromethyl)phenyl]methyl]-1H,2H,3H,4H,5H-cyclopenta[c]pyridine-2,7-dicarboxylate To a stirred solution of 5-tert-butyl 3-ethyl 1H,4H,5H,6H,7H-pyrazolo[4,3-c]pyridine-3,5-dicarboxylate (1.5 g, 5.079 mmol, 1 equiv.) and 1-(bromomethyl)-2-(trifluoromethyl)benzene (1.46 g, 6.095 mmol, 1.2 equiv.) in ACN (20 mL, 380.494 mmol), KCO (1.40 g, 10.158 mmol, 2 equiv.) and KI (0.84 g, 5.079 mmol, 1 equiv.) were added in small portions at room temperature under a nitrogen atmosphere. The resulting mixture was stirred at 80 °C under a nitrogen atmosphere for 2 hours. The reaction was monitored by LCMS. The resulting mixture was concentrated under reduced pressure. The resulting mixture was extracted with EtOAc (50 mL). The combined organic layers were washed with brine (3×100 mL) and dried over anhydrous NaSO. After filtration, the filtrate was concentrated under reduced pressure. The resulting mixture was used directly in the next step (E00692-127) without further purification.

[0432] Ethyl 1-[[2-(trifluoromethyl)phenyl]methyl]-1H,4H,5H,6H,7H-pyrazolo[4,3-c]pyridine-3-carboxylate To a stirred solution of 2-tert-butyl 7-ethyl 5-[[2-(trifluoromethyl)phenyl]methyl]-1H,2H,3H,4H,5H-cyclopenta[c]pyridine-2,7-dicarboxylate (1 g, 2.215 mmol, 1 equiv.) in DCM (10 mL) was added TFA (3 mL) in small portions at room temperature under a nitrogen atmosphere. The resulting mixture was stirred at room temperature under a nitrogen atmosphere for 2 hours. The reaction was monitored by LCMS. The resulting mixture was concentrated under reduced pressure. The resulting mixture was used directly in the next step (E00692-129) without further purification.

[0433] Ethyl 5-[5-chloro-1-(oxan-2-yl)-6-oxo-1,6-dihydropyridazin-4-yl]-1-[[2-(trifluoromethyl)phenyl]methyl]-1H,4H,5H,6H,7H-pyrazolo[4,3-c]pyridine-3-carboxylate To a stirred solution of ethyl 1-[[2-(trifluoromethyl)phenyl]methyl]-1H,4H,5H,6H,7H-pyrazolo[4,3-c]pyridine-3-carboxylate (750 mg, 2.123 mmol, 1 equiv.) and 4,5-dichloro-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (528.71 mg, 2.123 mmol, 1 equiv.) was added DIEA (5 mL) in small portions at room temperature under a nitrogen atmosphere. The resulting mixture was stirred as a neat reaction at 100° C. overnight under a nitrogen atmosphere. The reaction was monitored by LCMS. The mixture was allowed to cool to room temperature. The resulting mixture was extracted with EtOAc (30 mL). The combined organic layers were washed with brine (3×50 mL) and dried over anhydrous NaSO. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC (PE / EtOAc=1:1) to give ethyl 5-[5-chloro-1-(oxan-2-yl)-6-oxo-1,6-dihydropyridazin-4-yl]-1-[[2-(trifluoromethyl)phenyl]methyl]-1H,4H,5H,6H,7H-pyrazolo[4,3-c]pyridine-3-carboxylate (1 g, 83.24%) as a pale yellow oil.

[0434] 5-[5-chloro-1-(oxan-2-yl)-6-oxo-1,6-dihydropyridazin-4-yl]-1-[[2-(trifluoromethyl)phenyl]methyl]-1H,4H,5H,6H,7H-pyrazolo[4,3-c]pyridine-3-carboxylic acid To a stirred solution of ethyl 5-[5-chloro-1-(oxan-2-yl)-6-oxo-1,6-dihydropyridazin-4-yl]-1-[[2-(trifluoromethyl)phenyl]methyl]-1H,4H,5H,6H,7H-pyrazolo[4,3-c]pyridine-3-carboxylate (1 g, 1.767 mmol, 1 equiv.) in THF (5 mL) and HO (5 mL) was added LiOH (0.21 g, 0.009 mmol, 5 equiv.) in a nitrogen atmosphere at room temperature. The resulting mixture was stirred at 50° C. for 3 hours under a nitrogen atmosphere. The reaction was monitored by LCMS. The mixture was acidified to pH 6 with HCl (aq.). The resulting mixture was extracted with EtOAc (30 mL). The combined organic layers were washed with brine (3×10 mL) and dried over anhydrous NaSO. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with CHCl / MeOH (50:1 to 5:1) to give 5-[5-chloro-1-(oxan-2-yl)-6-oxo-1,6-dihydropyridazin-4-yl]-1-[[2-(trifluoromethyl)phenyl]methyl]-1H,4H,5H,6H,7H-pyrazolo[4,3-c]pyridine-3-carboxylic acid (700 mg, 73.65%) as a pale yellow oil.

[0435] 5-[5-chloro-1-(oxan-2-yl)-6-oxo-1,6-dihydropyridazin-4-yl]-1-[[2-(trifluoromethyl)phenyl]methyl]-1H,4H,5H,6H,7H-pyrazolo[4,3-c]pyridine-3-carboxamide To a stirred solution of 5-[5-chloro-1-(oxan-2-yl)-6-oxo-1,6-dihydropyridazin-4-yl]-1-[[2-(trifluoromethyl)phenyl]methyl]-1H,4H,5H,6H,7H-pyrazolo[4,3-c]pyridine-3-carboxylic acid (700 mg, 1.301 mmol, 1 equiv.) in DMF (10 mL) was added CDI (316.51 mg, 1.952 mmol, 1.5 equiv.) in portions at room temperature under a nitrogen atmosphere. The resulting mixture was stirred at 50° C. for 1 hour under a nitrogen atmosphere. To the above mixture was added NHOAc (300.92 mg, 3.904 mmol, 3 equiv.) in portions over 5 minutes at 50° C. The resulting mixture was stirred at 50° C. for an additional 2 hours. The reaction was monitored by LCMS. The mixture was allowed to cool to room temperature. The resulting mixture was extracted with EtOAc (30 mL). The combined organic layers were washed with brine (3 × 50 mL) and dried over anhydrous NaSO. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with PE / EtOAc (20:1 to 5:1) to afford 5-[5-chloro-1-(oxan-2-yl)-6-oxo-1,6-dihydropyridazin-4-yl]-1-[[2-(trifluoromethyl)phenyl]methyl]-1H,4H,5H,6H,7H-pyrazolo[4,3-c]pyridine-3-carboxamide (40 mg, 5.72%) as a pale yellow oil.

[0436] 5-(5-chloro-6-oxo-1,6-dihydropyridazin-4-yl)-1-[[2-(trifluoromethyl)phenyl]methyl]-1H,4H,5H,6H,7H-pyrazolo[4,3-c]pyridine-3-carboxamide To a stirred solution of 5-[5-chloro-1-(oxan-2-yl)-6-oxo-1,6-dihydropyridazin-4-yl]-1-[[2-(trifluoromethyl)phenyl]methyl]-1H,4H,5H,6H,7H-pyrazolo[4,3-c]pyridine-3-carboxamide (40 mg, 0.074 mmol, 1 equiv.) in DCM (10 mL) was added TFA (3 mL, 40.389 mmol, 542.16 equiv.) at room temperature. The resulting mixture was stirred at room temperature for 2 hours. The reaction was monitored by LCMS. The resulting mixture was concentrated under reduced pressure. The mixture was basified to pH 8 with saturated NaHCO3 (aq.). The crude product (30 mg) was purified by preparative HPLC using the following conditions: (Column = XBridge Shield RP18OBD column, 5 μm, 19 × 150 mm, Mobile phase A = water (10 mmol / L NH4HCO3), Mobile phase B = ACN, Flow rate = 20 mL / min, Gradient = 24% B to 45% B in 7 min, 220 / 254 nm, Rt = 6.45 min) to give 5-(5-chloro-6-oxo-1,6-dihydropyridazin-4-yl)-1-[[2-(trifluoropyrazolo)phenyl]methyl]-1H,4H,5H,6H,7H-pyrazolo[4,3-c]pyridine-3-carboxamide (12.5 mg, 37.06%) as a white solid.

[0437] Example 31 Synthesis of Compound NL

[0438] [ka] Preparation of NK and NL 1-Bromo-2-(difluoromethyl)-4-fluorobenzene To a stirred solution of 2-bromo-5-fluorobenzaldehyde (10 g, 49.26 mmol, 1 equiv) in DCM (10 mL) was added DAST (15.9 g, 98.64 mmol, 2.00 equiv). The resulting mixture was stirred at −10° C. for 2 hours. The resulting mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with PE / EtOAc (5:1) to give 1-bromo-2-(difluoromethyl)-4-fluorobenzene (8.2 g, 73.98%) as a pale yellow oil.

[0439] 2-(Difluoromethyl)-4-fluorobenzaldehyde A solution of 1-bromo-2-(difluoromethyl)-4-fluorobenzene (8 g, 35.55 mmol, 1 equiv.) and n-BuLi (2.7 g, 42.15 mmol, 1.19 equiv.) in THF (150 mL) was stirred at −78° C. for 2 h. To the above mixture was added DMF (3.9 g, 53.33 mmol, 1.5 equiv.). The resulting mixture was stirred at −78° C. for 1 h. The reaction was quenched by adding water (50 mL) at −70° C. The solution was extracted with EtOAc (3×50 mL). The combined organic layers were washed with brine (2×30 mL) and dried over anhydrous NaSO. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with PE / EtOAc (5:1) to give 2-(difluoromethyl)-4-fluorobenzaldehyde (3 g, 48.46%) as a pale yellow oil.

[0440] 1-[2-(difluoromethyl)-4-fluorophenyl]ethan-1-ol To a stirred solution of 2-(difluoromethyl)-4-fluorobenzaldehyde (3 g, 17.23 mmol, 1 equiv) in THF (30 mL, 416.06 mmol, 10 equiv) was added CHMgBr (25.84 mL, 25.84 mmol, 1.5 equiv) dropwise at −30° C. under a nitrogen atmosphere. The resulting mixture was stirred at −10° C. for 2 h under a nitrogen atmosphere. The reaction was quenched with saturated NHCl (aq) at 0° C. The mixture was extracted with EtOAc (3×300 mL). The combined organic layers were washed with brine (3×300 mL) and dried over anhydrous NaSO. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with PE / EtOAc (100:1 to 50:1) to give 1-[2-(difluoromethyl)-4-fluorophenyl]ethan-1-ol (2.68 g, 81.80%) as a red oil.

[0441] 1-(1-chloroethyl)-2-(difluoromethyl)-4-fluorobenzene To a stirred solution / mixture of 1-[2-(difluoromethyl)-4-fluorophenyl]ethan-1-ol (2.68 g, 14.09 mmol, 1 equiv) in DCM (30 mL, 140.93 mmol, 10 equiv) was added SO2Cl2 (6.7 g, 49.64 mmol, 3.52 equiv) dropwise under air atmosphere at 0° C. The resulting mixture was stirred at 20° C. for 2 h and the resulting oil was dried under vacuum to give 1-(1-chloroethyl)-2-(difluoromethyl)-4-fluorobenzene (2.36 g, 80.27%) as a red oil.

[0442] 1-[2-(difluoromethyl)-4-fluorophenyl]ethan-1-amine To a stirred solution of 1-(1-chloroethyl)-2-(difluoromethyl)-4-fluorobenzene (300 mg, 1.44 mmol, 1 equiv.) in MeOH was added NH3 (g) at room temperature under a nitrogen atmosphere. The resulting mixture was stirred at 70° C. for 20 hours under a nitrogen atmosphere. The reaction was monitored by LCMS. The mixture was allowed to cool to room temperature. The resulting mixture was concentrated under reduced pressure. This gave 1-[2-(difluoromethyl)-4-fluorophenyl]ethan-1-amine (130 mg, 47.78%) as a yellow oil. The resulting mixture was used directly in the next step without further purification.

[0443] 4-chloro-5-[1-[(1S)-1-[2-(difluoromethyl)-4-fluorophenyl]ethyl]-1H,4H,5H,6H,7H-[1,2,3]triazolo[4,5-c]pyridin-5-yl]-2,3-dihydropyridazin-3-one and 4-chloro-5-[1-[(1R)-1-[2-(difluoromethyl)-4-fluorophenyl]ethyl]-1H,4H,5H,6H,7H-[1,2,3]triazolo[4,5-c]pyridin-5-yl]-2,3-dihydropyridazin-3-one To a stirred mixture of 1-[2-(difluoromethyl)-4-fluorophenyl]ethan-1-amine (130.0 mg, 0.69 mmol, 2.00 equiv.) and 4-chloro-5-(4-oxopiperidin-1-yl)-2,3-dihydropyridazin-3-one (78.2 mg, 0.34 mmol, 1 equiv.) in DMF (10 mL), 1-azido-4-nitrobenzene (78.9 mg, 0.48 mmol, 1.40 equiv.) and Zn(OAc) (63.0 mg, 0.34 mmol, 1.00 equiv.) were added in small portions at room temperature under a nitrogen atmosphere. The resulting mixture was stirred at 60° C. for 16 hours under a nitrogen atmosphere. The reaction was monitored by LCMS. The mixture was allowed to cool to room temperature. The residue was purified by reverse-phase flash chromatography using the following conditions: (Column: XBridge Shield RP18OBD column, 20-40 μm, 19 × 150 mm; Mobile phase A: water (10 mmol / L NH4HCO3), Mobile phase B: ACN, Flow rate: 80 mL / min, Gradient: 30% B to 70% B in 30 min, 220 nm, Rt = 7.08 min) to give a mixture of products. The residue (100 mg) was purified by chiral preparative HPLC using the following conditions: Column: CHIRALPAK IF-3, 0.46 × 5 cm, 3 μm, Mobile phase: MtBE (0.1% DEA):EtOH = 80:20, Detector: UV-254 nm. 4-Chloro-5- [1- [(1S)-1- [2- (difluoromethyl) -4-fluorophenyl] ethyl] -1H, 4H, 5H, 6H, 7H- [1,2,3] triazolo [4,5-c] pyridin-5-yl] -2,3-dihydropyridazin-3-one (19.0 mg) was obtained as an off-white solid in 3.835 minutes, 4-chloro-5- [1- [(1R)-1- [2- (difluoromethyl) -4-fluorophenyl] ethyl] -1H, 4H, 5H, 6H, 7H- [1,2,3] triazolo [4,5-c] pyridin-5-yl] -2,3-dihydropyridazin-3-one (33.8 mg) was obtained as an off-white solid in 3.185 minutes.

[0444] Example 32 Synthesis of Compound QM

[0445] [ka] Compound QM was prepared by using the corresponding aniline according to the method and scheme described for QL.

[0446] [ka] Preparation of QL 2-ethenyl-3-nitropyridine To a stirred mixture of 2-chloro-3-nitropyridine (2 g, 12.615 mmol, 1 equiv.) and NaCO (2.67 g, 25.230 mmol, 2.0 equiv.) in 1,4-dioxane (20 mL) and HO (1 mL), Pd(PPh) (0.73 g, 0.631 mmol, 0.05 equiv.) and 2-ethenyl-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (1.94 g, 12.615 mmol, 1.00 equiv.) were added under a nitrogen atmosphere at 0 °C. The resulting mixture was stirred under a nitrogen atmosphere at room temperature for 3 h. The reaction was monitored by LCMS. The resulting mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with PE / EtOAc (10:1 to 5:1) to give 2-ethenyl-3-nitropyridine (1.1 g, 58.08%) as a brown solid.

[0447] 2-Ethylpyridin-3-amine To a stirred solution of 2-ethenyl-3-nitropyridine (1.1 g, 7.327 mmol, 1 equiv.) in MeOH (10 mL) was added Pd / C (100 mg, 0.266 mmol, 0.04 equiv.) at room temperature under a hydrogen atmosphere. The resulting mixture was stirred at room temperature under a hydrogen atmosphere for 16 hours. The reaction was monitored by LCMS. The resulting mixture was filtered, and the filter cake was washed with MeOH (2×10 mL). The filtrate was concentrated under reduced pressure. The residual product was purified by reverse-phase flash using the following conditions: (Column = XBridge Prep OBD C18 column, 30 × 150 mm, 5 μm, Mobile phase A = water (10 mmol / L NH4HCO3), Mobile phase B = ACN, Flow rate = 60 mL / min, Gradient = 20% B to 40% B in 11 min, 220 nm, Rt = 11.77 min) to give 2-ethylpyridin-3-amine (620 mg, 69.27%) as a white solid.

[0448] tert-Butyl 2-[(2-ethylpyridin-3-yl)amino]-5H,6H,7H-pyrrolo[3,4-d]pyrimidine-6-carboxylate To a stirred mixture of tert-butyl 2-chloro-5H,6H,7H-pyrrolo[3,4-d]pyrimidine-6-carboxylate (200 mg, 0.782 mmol, 1 equiv.) and 2-ethyl-3-nitropyridine (238.02 mg, 1.564 mmol, 2.0 equiv.) in 1,4-dioxane (20 mL), CsCO (509.69 mg, 1.564 mmol, 2.0 equiv.) and Pd(AcO) (35.12 mg, 0.156 mmol, 0.2 equiv.) were added at room temperature under a nitrogen atmosphere. Xantphos (181.03 mg, 0.313 mmol, 0.4 equiv.) was then added at room temperature under a nitrogen atmosphere. The final reaction mixture was irradiated at 110 °C for 2 h by microwave irradiation. The reaction was monitored by LCMS. The mixture was allowed to cool to room temperature. The resulting mixture was filtered, and the filter cake was washed with CHCl (2×10 mL). The filtrate was concentrated under reduced pressure. The crude product was purified by reverse-phase flash using the following conditions: (Column: XBridge Prep OBD C18 column, 30×150 mm, 5 μm, Mobile Phase A: water (10 mmol / L NHHCO), Mobile Phase B: ACN, Flow Rate: 60 mL / min, Gradient: 20% B to 40% B in 11 min, 220 nm, Rt = 11.77 min) to give tert-butyl 2-[(2-ethylpyridin-3-yl)amino]-5H,6H,7H-pyrrolo[3,4-d]pyrimidine-6-carboxylate (250 mg, 93.62%) as a brown solid.

[0449] tert-Butyl 2-[(2-ethylpyridin-3-yl)(methyl)amino]-5H,6H,7H-pyrrolo[3,4-d]pyrimidine-6-carboxylate To a stirred solution of tert-butyl 2-[(2-ethylpyridin-3-yl)amino]-5H,6H,7H-pyrrolo[3,4-d]pyrimidine-6-carboxylate (300 mg, 0.879 mmol, 1 equiv.) in DMF (10 mL) was added NaH (42.17 mg, 1.757 mmol, 2.0 equiv.) at 0° C. under a nitrogen atmosphere. The resulting mixture was stirred at 0° C. under a nitrogen atmosphere for 1 hour. Then, CHCl (249.44 mg, 1.757 mmol, 2.00 equiv.) was added at 0° C. under a nitrogen atmosphere. The resulting mixture was stirred at room temperature under a nitrogen atmosphere for 1 hour. The reaction was monitored by LCMS. The resulting mixture was diluted with water (2 mL). The crude product was purified by reverse-phase flash using the following conditions: (Column = XBridge Prep OBD C18 column, 30 × 150 mm, 5 μm, Mobile phase A = water (10 mmol / L NH4HCO3), Mobile phase B = ACN, Flow rate = 60 mL / min, Gradient = 20% B to 40% B in 11 min, 220 nm, Rt = 11.77 min) to give tert-butyl 2-[(2-ethylpyridin-3-yl)(methyl)amino]-5H,6H,7H-pyrrolo[3,4-d]pyrimidine-6-carboxylate (250 mg, 80.04%) as a brown solid.

[0450] N-(2-ethylpyridin-3-yl)-N-methyl-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-2-amine To a stirred solution of tert-butyl 2-[(2-ethylpyridin-3-yl)(methyl)amino]-5H,6H,7H-pyrrolo[3,4-d]pyrimidine-6-carboxylate (200 mg, 0.586 mmol, 1 equiv.) in DCM (4 mL) was added TFA (1 mL, 13.463 mmol, 22.98 equiv.) at room temperature. The resulting mixture was stirred at room temperature for 1 hour. The reaction was monitored by LCMS. The resulting mixture was concentrated under reduced pressure. The mixture was basified to pH 8 with saturated NaHCO3 (aq.). The crude product was purified by reverse-phase flash using the following conditions: (Column = XBridge Prep OBD C18 column, 30 × 150 mm, 5 μm, Mobile phase A = water (10 mmol / L NH4HCO3), Mobile phase B = ACN, Flow rate = 60 mL / min, Gradient = 18% B to 35% B in 8 min, 220 nm, Rt = 7.12 min) to give N-(2-ethylpyridin-3-yl)-N-methyl-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-2-amine (120 mg, 84.89%) as a brown solid.

[0451] 4-chloro-5-[2-[(2-ethylpyridin-3-yl)(methyl)amino]-5H,6H,7H-pyrrolo[3,4-d]pyrimidin-6-yl]-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one To a stirred solution of N-(2-ethylpyridin-3-yl)-N-methyl-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-2-amine (120 mg, 0.497 mmol, 1 equiv.) in DIEA (0.1 mL) was added 4,5-dichloro-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (99.10 mg, 0.398 mmol, 0.80 equiv.) at room temperature. The resulting mixture was stirred at 90° C. for 1 hour. The reaction was monitored by LCMS. The mixture was allowed to cool to room temperature. The resulting mixture was concentrated under reduced pressure. The residue was purified by preparative TLC (CHCl / MeOH=12:1) to give 4-chloro-5-[2-[(2-ethylpyridin-3-yl)(methyl)amino]-5H,6H,7H-pyrrolo[3,4-d]pyrimidin-6-yl]-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (100 mg, 42.97%) as a brown solid.

[0452] 4-chloro-5-[2-[(2-ethylpyridin-3-yl)(methyl)amino]-5H,6H,7H-pyrrolo[3,4-d]pyrimidin-6-yl]-2,3-dihydropyridazin-3-one To a stirred solution of 4-chloro-5-[2-[(2-ethylpyridin-3-yl)(methyl)amino]-5H,6H,7H-pyrrolo[3,4-d]pyrimidin-6-yl]-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (100 mg, 0.214 mmol, 1 equiv.) in DCM (4 mL) was added TFA (1 mL) at room temperature. The resulting mixture was stirred at room temperature for 1 hour. The reaction was monitored by LCMS. The resulting mixture was concentrated under reduced pressure. The mixture was basified to pH 8 with saturated NaHCO3 (aq.). The crude product (80 mg) was purified by preparative HPLC using the following conditions: (Column = XBridge Prep OBD C18 column, 30 × 150 mm, 5 μm, Mobile phase A = water (10 mmol / L NH4HCO3), Mobile phase B = ACN, Flow rate = 60 mL / min, Gradient = 15% B to 35% B in 8 min, 220 nm, Rt = 6.65 min) to give 4-chloro-5-[2-[(2-ethylpyridin-3-yl)(methyl)amino]-5H,6H,7H-pyrrolo[3,4-d]pyrimidin-6-yl]-2,3-dihydropyridazin-3-one (67.4 mg, 82.17%) as a white solid.

[0453] Example 33 Synthesis of Compounds MD and ME

[0454] [ka] Step 1. tert-Butyl N-[(2R)-1-(2-chloroacetamido)propan-2-yl]carbamate To a stirred solution of tert-butyl N-[(2R)-1-aminopropan-2-yl]carbamate (3 g, 17.217 mmol, 1 equiv.) in EA (50 mL) was added a solution of Na2CO3 (3649.65 mg, 34.434 mmol, 2 equiv.) in HO (10 mL) at room temperature. Then, a solution of 2-chloroacetyl chloride (3.89 g, 34.434 mmol, 2 equiv.) in EA (10 mL) was added dropwise at 0 °C. The resulting mixture was stirred at room temperature for 2 h. The reaction was monitored by LCMS. The reaction was quenched with water at room temperature. The resulting mixture was extracted with EtOAc (3 × 100 mL). The combined organic layers were washed with brine (1 × 100 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. This gave tert-butyl N-[(2R)-1-(2-chloroacetamido)propan-2-yl]carbamate (4.5 g, crude) as a white solid.

[0455] Step 2. (5R)-5-Methylpiperazin-2-one To a stirred solution of tert-butyl N-[(2R)-1-(2-chloroacetamido)propan-2-yl]carbamate (4.5 g, 17.948 mmol, 1 equiv) in DCM (30 mL) was added a solution of TFA (10 mL, 134.630 mmol, 7.50 equiv) in DCM (10 mL) dropwise at 0 °C. The resulting mixture was stirred at room temperature for 2 h. The reaction was monitored by LCMS. The resulting mixture was concentrated under reduced pressure. To the above mixture, K2CO3 (4.96 g, 35.897 mmol, 2 equiv) and KI (2.98 g, 17.948 mmol, 1 equiv) were added at room temperature. The resulting mixture was stirred at 80 °C for an additional 16 h. The reaction was monitored by LCMS. The resulting mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with CH2Cl2 / MeOH (20:1 to 10:1) to give (5R)-5-methylpiperazin-2-one (2.5 g, crude) as a yellow oil.

[0456] Step 3. 4-chloro-5-[(2R)-2-methyl-5-oxopiperazin-1-yl]-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one To a stirred solution of (5R)-5-methylpiperazin-2-one (2.5 g, 21.901 mmol, 1 equiv.) in DIEA (2 mL) was added 4,5-dichloro-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (5.46 g, 21.901 mmol, 1 equiv.) at room temperature. The resulting mixture was stirred at 100° C. for 16 hours. The reaction was monitored by LCMS. The mixture was allowed to cool to room temperature. The residue was purified by reverse-phase flash using the following conditions: (Column = C18 330 g, Mobile phase A = water / 0.05% NH4HCO3, Mobile phase B = ACN, Flow rate = 80 mL / min, Gradient = 20% B to 30% B in 10 min, Detector = 220 nm, Monitor = 254 nm) to give 4-chloro-5-[(2R)-2-methyl-5-oxopiperazin-1-yl]-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (600 mg, 8.38%) as a yellow solid.

[0457] Step 4. 4-chloro-5-[(2R)-4-[1-[4-fluoro-2-(trifluoromethyl)phenyl]ethyl]-2-methyl-5-oxopiperazin-1-yl]-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one To a stirred mixture of 4-chloro-5-[(2R)-2-methyl-5-oxopiperazin-1-yl]-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (500 mg, 1.530 mmol, 1 equiv.) and 1-[4-fluoro-2-(trifluoromethyl)phenyl]ethyl methanesulfonate (656.96 mg, 2.295 mmol, 1.5 equiv.) in ACN (20 mL) was added t-BuONa (220.57 mg, 2.295 mmol, 1.5 equiv.) at room temperature under a nitrogen atmosphere. The final reaction mixture was irradiated by microwave irradiation at 110° C. for 3 hours. The reaction was monitored by LCMS. The mixture was allowed to cool to room temperature. The resulting mixture was concentrated under reduced pressure. The residue was purified by reverse-phase flash using the following conditions: (Column = C18, 330 g, Mobile phase A = water / 0.05% NH4HCO3, Mobile phase B = ACN, Flow rate = 80 mL / min, Gradient = 55% B to 75% B in 15 min, Detector = 220 nm, Monitor = 254 nm) to give 4-chloro-5-[(2R)-4-[1-[4-fluoro-2-(trifluoromethyl)phenyl]ethyl]-2-methyl-5-oxopiperazin-1-yl]-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (120 mg, 15.17%) as a yellow solid.

[0458] Step 5. MD and ME 4-chloro-5-[(2R)-4-[(1S)-1-[4-fluoro-2-(trifluoromethyl)phenyl]ethyl]-2-methyl-5-oxopiperazin-1-yl]-2,3-dihydropyridazin-3-one and 4-chloro-5-[(2R)-4-[(1R)-1-[4-fluoro-2-(trifluoromethyl)phenyl]ethyl]-2-methyl-5-oxopiperazin-1-yl]-2,3-dihydropyridazin-3-one To a stirred solution of 4-chloro-5-[(2R)-4-[1-[4-fluoro-2-(trifluoromethyl)phenyl]ethyl]-2-methyl-5-oxopiperazin-1-yl]-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (120 mg, 0.232 mmol, 1 equiv.) in DCM (8 mL) was added TFA (2 mL, 26.926 mmol, 115.99 equiv.) at room temperature. The resulting mixture was stirred at room temperature for 1 hour. The reaction was monitored by LCMS. The resulting mixture was concentrated under reduced pressure. The residue was basified to pH 8 with saturated NaHCO3 (aq.). The resulting mixture was concentrated under reduced pressure. The residue was purified using the following conditions: (Column = XBridge Prep C18 Purification was performed by reversed-phase flash using an OBD column, 19 x 150 mm, 5 um, mobile phase A = water (10 mmol / L NH4HCO3), mobile phase B = ACN, flow rate = 60 mL / min, gradient = 22% B to 51% B in 7 min, 254 / 220 nm, Rt = 6.4 min) with 4-chloro-5-[(2R)-4-[(1S)-1-[4-fluoro-2-(trifluoromethyl)phenyl]ethyl]-2-methyl-5 -oxopiperazin-1-yl]-2,3-dihydropyridazin-3-one (16.3 mg, 16.22%) as a white solid, and 4-chloro-5-[(2R)-4-[(1R)-1-[4-fluoro-2-(trifluoromethyl)phenyl]ethyl]-2-methyl-5-oxopiperazin-1-yl]-2,3-dihydropyridazin-3-one (18.6 mg, 18.51%) as a white solid.

[0459] Example 34 Synthesis of Compound MF

[0460] [ka] Step 1. 1-[4-fluoro-2-(trifluoromethyl)phenyl]ethan-1-ol To a stirred solution of 4-fluoro-2-(trifluoromethyl)benzaldehyde (3 g, 15.616 mmol, 1 equiv) in THF (50 mL) was added MeMgBr in EtO (3 mol / L, 30 ml) dropwise at −30° C. under a nitrogen atmosphere. The resulting mixture was stirred at room temperature under a nitrogen atmosphere for 2 hours. The reaction was monitored by TLC. The reaction was quenched with saturated NH4Cl (aq) at 0° C. The resulting mixture was extracted with EtOAc (50 mL). The combined organic layers were washed with brine (3×50 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The resulting mixture was used directly in the next step without further purification.

[0461] Step 2. 1-[4-Fluoro-2-(trifluoromethyl)phenyl]ethyl methanesulfonate To a stirred solution of 1-[4-fluoro-2-(trifluoromethyl)phenyl]ethan-1-ol (3 g, 14.412 mmol, 1 equiv.) and EtN (2.92 g, 28.825 mmol, 2 equiv.) in DCM (60 mL) was added MsCl (2.48 g, 21.618 mmol, 1.5 equiv.) dropwise at 0° C. under a nitrogen atmosphere. The resulting mixture was stirred at room temperature under a nitrogen atmosphere for 2 h. The reaction was monitored by TLC. The reaction was quenched with saturated NH4Cl(aq.) (50 mL) at 0° C. The resulting mixture was extracted with EtOAc (50 mL). The combined organic layers were washed with brine (3×100 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure to give 1-[4-fluoro-2-(trifluoromethyl)phenyl]ethyl methanesulfonate (1.6 g, 38.78%) as a pale yellow oil.

[0462] Step 3. 4-chloro-5-(4-[1-[4-fluoro-2-(trifluoromethyl)phenyl]ethyl]-3-oxopiperazin-1-yl)-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one To a stirred solution of 4-chloro-2-(oxan-2-yl)-5-(3-oxopiperazin-1-yl)-2,3-dihydropyridazin-3-one (800 mg, 2.558 mmol, 1 equiv.) and 1-[4-fluoro-2-(trifluoromethyl)phenyl]ethyl methanesulfonate (878.63 mg, 3.070 mmol, 1.2 equiv.) in ACN (8 mL) was added sodium 2,2-dimethylpropan-1-olate (563.43 mg, 5.116 mmol, 2 equiv.) in a nitrogen atmosphere at room temperature in small portions. The final reaction mixture was irradiated by microwave irradiation at 110 °C for 3 h. The reaction was monitored by LCMS. The mixture was allowed to cool to room temperature. The resulting mixture was extracted with EtOAc (20 mL). The combined organic layers were washed with brine (3 × 10 mL) and dried over anhydrous NaSO. After filtration, the filtrate was concentrated under reduced pressure, and the resulting mixture was used directly in the next step without further purification.

[0463] Step 4. Compound MF 4-chloro-5-[4-[(1R)-1-[4-fluoro-2-(trifluoromethyl)phenyl]ethyl]-3-oxopiperazin-1-yl]-2,3-dihydropyridazin-3-one and 4-chloro-5-[4-[(1S)-1-[4-fluoro-2-(trifluoromethyl)phenyl]ethyl]-3-oxopiperazin-1-yl]-2,3-dihydropyridazin-3-one To a stirred solution of 4-chloro-5-(4-[1-[4-fluoro-2-(trifluoromethyl)phenyl]ethyl]-3-oxopiperazin-1-yl)-2-(oxan-2-yl)-2,3-dihydropyridazin-3-one (110 mg, 0.219 mmol, 1 equiv) in DCM (10 mL) was added TFA (3 mL) in small portions at room temperature under a nitrogen atmosphere. The resulting mixture was stirred at room temperature under a nitrogen atmosphere for 3 hours. The reaction was monitored by LCMS. The resulting mixture was concentrated in vacuo. The residue was basified to pH 8 with saturated NaHCO3 (aq). The resulting mixture was extracted with EtOAc (20 mL). The combined organic layers were washed with brine (3 x 20 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The crude product (50 mg) was purified using the following conditions: (Column: XBridge Prep OBD Purified by chiral HPLC using a C18 column, 30 x 150 mm, 5 µm, mobile phase A = water (10 mmol / L NH4HCO3), mobile phase B = ACN, flow rate = 60 mL / min, gradient = 20% B to 32% B in 16 min, 220 nm, Rt = 14.27 min) and 4-chloro-5-[4-[(1R)-1-[4-fluoro-2-(trifluoromethyl)phenyl]ethyl]- 3-Oxopiperazin-1-yl]-2,3-dihydropyridazin-3-one (6.0 mg, 6.55%) as a white solid and 4-chloro-5-[4-[(1S)-1-[4-fluoro-2-(trifluoromethyl)phenyl]ethyl]-3-oxopiperazin-1-yl]-2,3-dihydropyridazin-3-one (6.2 mg, 6.77%) as a white solid.

[0464] Example 35 Synthesis of Compound PW

[0465] [ka] Compound PW was prepared by the method and scheme described for PU using the corresponding amine.

[0466] [ka] Preparation of PU Preparation of Intermediate 9 (Int9) 1,5-Di-tert-butyl 1H,4H,5H,6H,7H-imidazo[4,5-c]pyridine-1,5-dicarboxylate To a stirred solution of 1H,4H,5H,6H,7H-imidazo[4,5-c]pyridine dihydrochloride (22 g, 112.20 mmol, 1 equiv.) in MeOH (300 mL), di-tert-butyl decarbonate (61.2 g, 280.50 mmol, 2.5 equiv.) and ethyl bis(propan-2-yl)amine (50.8 g, 392.70 mmol, 3.5 equiv.) were added dropwise at 0° C. under a nitrogen atmosphere. The solution was stirred at room temperature overnight. The desired product could be detected by LCMS. The mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with PE / EtOAc (5:1 to 2:1) to give 1,5-di-tert-butyl 1H,4H,5H,6H,7H-imidazo[4,5-c]pyridine-1,5-dicarboxylate (30 g, 82.68%) as a white solid.

[0467] To a stirred solution of 1,5-di-tert-butyl 1H,4H,5H,6H,7H-imidazo[4,5-c]pyridine-1,5-dicarboxylate (7 g, 1 equiv.) in MeOH (80 mL) and HO (17 mL), NaOH (1.7 g, 43.29 mmol, 2.00 equiv.) was added portionwise at room temperature under a nitrogen atmosphere. The mixture was stirred at room temperature for 2 hours. The desired product could be detected by LCMS. The mixture was basified to pH 8 with citric acid. The resulting mixture was extracted with CHCl (3 × 100 mL). The combined organic layers were washed with brine (1 × 100 mL) and dried over anhydrous NaSO. After filtration, the filtrate was concentrated under reduced pressure to give tert-butyl 1H,4H,5H,6H,7H-imidazo[4,5-c]pyridine-5-carboxylate (4.1 g, 84.84%) as an off-white semi-solid.

[0468] tert-Butyl 1-[[2-(trifluoromethyl)phenyl]methyl]-1H,4H,5H,6H,7H-imidazo[4,5-c]pyridine-5-carboxylate To a stirred solution of tert-butyl 1H,4H,5H,6H,7H-imidazo[4,5-c]pyridine-5-carboxylate (93.4 mg, 0.42 mmol, 1 equiv.) in DMF (8 mL) was added NaH (25.1 mg, 0.63 mmol, 1.5 equiv., 60%) in portions at 0° C. under a nitrogen atmosphere. The mixture was stirred at room temperature for 1 hour. To the mixture was added 1-(bromomethyl)-2-(trifluoromethyl)benzene (100 mg, 0.42 mmol, 1 equiv.) at 0° C. The mixture was stirred at room temperature for 1 hour. The desired product could be detected by LCMS. This was a test reaction and no workup was performed.

[0469] tert-Butyl 2-bromo-1-[[2-(trifluoromethyl)phenyl]methyl]-1H,4H,5H,6H,7H-imidazo[4,5-c]pyridine-5-carboxylate To a stirred solution of tert-butyl 1-[[2-(trifluoromethyl)phenyl]methyl]-1H,4H,5H,6H,7H-imidazo[4,5-c]pyridine-5-carboxylate (1 g, 2.62 mmol, 1 equiv.) in DMF (15 mL) was added NBS (0.5 g, 2.81 mmol, 1.07 equiv.) in portions at 0° C. under a nitrogen atmosphere. The mixture was stirred at room temperature for 1 hour. The desired product could be detected by LCMS. The resulting mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with PE / EtOAc (5:1 to 1:1) to give tert-butyl 2-bromo-1-[[2-(trifluoromethyl)phenyl]methyl]-1H,4H,5H,6H,7H-imidazo[4,5-c]pyridine-5-carboxylate (800 mg, 66.29%) as a colorless oil.

[0470] 5-tert-butyl 2-methyl 1-[[2-(trifluoromethyl)phenyl]methyl]-1H,4H,5H,6H,7H-imidazo[4,5-c]pyridine-2,5-dicarboxylate To a stirred mixture of tert-butyl 2-bromo-1-[[2-(trifluoromethyl)phenyl]methyl]-1H,4H,5H,6H,7H-imidazo[4,5-c]pyridine-5-carboxylate (1 g, 2.173 mmol, 1 equiv.) and TEA (0.44 g, 4.348 mmol, 2.00 equiv.) in MeOH (100 mL) was added Pd(PPh3)4 (0.25 g, 0.217 mmol, 0.1 equiv.) at room temperature under a nitrogen atmosphere. The resulting mixture was stirred at 100 °C under a nitrogen atmosphere for 1 hour. The reaction was monitored by LCMS. The mixture was allowed to cool to room temperature. The resulting mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with PE / EtOAc (10:1 to 1:1) to give 5-tert-butyl 2-methyl-1-[[2-(trifluoromethyl)phenyl]methyl]-1H,4H,5H,6H,7H-imidazo[4,5-c]pyridine-2,5-dicarboxylate (800 mg, 83.80%) as a brown solid.

[0471] Methyl 1-(2-(trifluoromethyl)benzyl)-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-2-carboxylate To a stirred solution of 5-tert-butyl 2-methyl 1-[[2-(trifluoromethyl)phenyl]methyl]-1H,4H,5H,6H,7H-imidazo[4,5-c]pyridine-2,5-dicarboxylate (350 mg, 0.71 mmol, 1 equiv.) in DCM (12 mL) was added TFA (2 mL, 26.93 mmol, 37.81 equiv.) at room temperature. The solution was stirred at room temperature for 2 hours....

Claims

1. 1. A method of treating renal disease, comprising co-administering to a subject in need thereof: a. A TRPC5 inhibitor compound represented by the structural formula (A), or a tautomer or a pharma- ceutically acceptable salt thereof 【Chemistry 1】 (In the formula, Each R is independently H, alkyl, alkenyl, alkynyl, aryl, heterocyclyl, heteroaryl, halogen, -OH, CN, cycloalkyl, -O-alkyl, -O-cycloalkyl, -O-aryl, -aryl-O-aryl, -CF ３ , -C(H)F ２ , alkylene-CF ３ , alkylene-C(H)F ２ , -SO ２ -alkyl, -O-alkylene-O-alkyl, -heterocyclyl-L-R ４ and heteroaryl-L-R ４ is selected from the group consisting of R ４ is absent, alkyl, cycloalkyl, polycyclyl, aryl, heterocyclyl, heteroaryl, -C(O)N(R ５ ) ２ , and C.F. ３ is selected from the group consisting of R ５ is independently H or alkyl; R ６ is an alkyl, cycloalkyl, aryl, heterocyclyl, heteroaryl, alkylene-aryl, -C(O)N(R ５ ) ２ , and C.F. ３ is selected from the group consisting of L is absent or is methylene, -C(O)-, -SO ２ --, --CH ２ N(Me)-, -N(R ５ ) (R ６ ) -, -C(R ５ ) (R ６ )- and -O-R ６ is selected from the group consisting of Only one R is -heterocyclyl-L-R ４ or -heteroaryl-L-R ４ ) and b. a second therapeutic agent selected from the following: an immunomodulator, a calcineurin inhibitor, a renin-angiotensin-aldosterone system inhibitor, an antiproliferative agent, an alkylating agent, a corticosteroid, an angiotensin-converting enzyme inhibitor, an adrenocorticotropic hormone stimulator, an angiotensin receptor blocker, a sodium-glucose transport protein 2 inhibitor, a dual sodium-glucose transport protein 1 / 2 inhibitor, a nuclear factor-1 (erythroid-derived 2)-like 2 agonist, a chemokine receptor 2 inhibitor, a chemokine receptor 5 inhibitor, an endothelin 1 receptor antagonist, a beta blocker, a mineralocorticoid receptor antagonist, a loop or thiazide diuretic, a calcium channel blocker, a statin, a short-, intermediate- or long-acting insulin, a dipeptidyl peptidase 4 inhibitor, a glucagon-like peptide 1 receptor agonist, a sulfonylurea, an apoptosis signal-regulating kinase-1, a chymase inhibitor, or a selective glycation inhibitor proteins, renin inhibitors, interleukin-33 inhibitors, farnesoid X receptor agonists, soluble guanylate cyclase stimulators, thromboxane receptor antagonists, xanthine oxidase inhibitors, erythropoietin receptor agonists, cannabinoid receptor type 1 inverse agonists, NADPH oxidase inhibitors, anti-vascular endothelial growth factor B, anti-fibrotic agents, neprilysin inhibitors, dual CD80 / CD86 inhibitors, CD40 antagonists, cell collagenase inhibitors, Terol and lipid blockers, PDGFR antagonists, Slit guidance ligand 2, APOL1 inhibitors, Nrl2 activators / NF-κB inhibitors, somatostatin receptor agonists, PPAR gamma agonists, AMP-activated protein kinase stimulators, tyrosine kinase inhibitors, glucosylceramide synthase inhibitors, arginine vasopressin receptor 2 antagonists, xanthine oxidase inhibitors, and vasopressin receptor 2 antagonists.

2. The method according to claim 1, wherein the TRPC5 inhibitor compound is represented by structural formula (AI), (A-II), or (A-III), or a tautomer or a pharma- ceutically acceptable salt thereof. 【Chemistry 2】 (In the formula, R １ and R ３ are independently H, alkyl, alkenyl, alkynyl, aryl, heterocyclyl, heteroaryl, halogen, -OH, CN, cycloalkyl, -O-alkyl, -O-cycloalkyl, -O-aryl, -aryl-O-aryl, -CF ３ , -C(H)F ２ , alkylene-CF ３ , alkylene-C(H)F ２ , -SO ２ -alkyl, and -O-alkylene-O-alkyl, -heterocyclyl-L-R ４ and heteroaryl-L-R ４ is selected from the group consisting of R ２ Heterocyclyl-L-R ４ and R ４ is absent or is alkyl, cycloalkyl, aryl, alkylene-aryl, alkylene-heteroaryl, heteroaryl, heterocyclyl, —C(O)N(R ５ ) ２ , and C.F. ３ is selected from the group consisting of R ５ is independently H or alkyl; R ６ is an alkyl, cycloalkyl, aryl, heterocyclyl, heteroaryl, alkylene-aryl, -C(O)N(R ５ ) ２ , and C.F. ３ is selected from the group consisting of L is absent or is methylene, -C(O)-, -SO ２ --, --CH ２ N(Me)-, -N(R ５ ) (R ６ ) -, -C(R ５ ) (R ６ )- and -O-R ６ is selected from the group consisting of R １ , R ２ , and R ３ Exactly one of the groups is -heterocyclyl-L-R ４ or -heteroaryl-L-R ４ (It is.)

3. The TRPC5 inhibitor compound has the structural formula (I): 【Chemistry 3】 10. The method of claim 1 , comprising administering to said patient a compound of formula: "---" is a single bond or a double bond, X １ is CH or N, When "---" is a double bond, X ２ is CH or N, When "---" is a single bond, X ２ is N(CH ３ ) and X １ When is CH, X ２ is N or N(CH ３ ) and Y is -O-, -N(CH ３ ) -, -N(CH ２ CH ２ OH)-, cyclopropane-1,1-diyl, or -CH(CH ３ ) - and Q is 2-trifluoromethyl-4-fluorophenyl, 2-difluoromethyl-4-fluorophenyl, 2-trifluoromethylphenyl, 2-methyl-4-fluorophenyl, 2-chloro-4-fluorophenyl, 2-chlorophenyl, 1-(benzyl)-4-methylpiperidin-3-yl, 4-trifluoromethylpyridin-3-yl, 2-trifluoromethyl-6-fluorophenyl, 2-trifluoromethyl-3-cyanophenyl, 2-ethyl-3-fluorophenyl, 2-chloro-3-cyanophenyl, 2-trifluoromethyl-5-fluorophenyl, or 2-difluoromethylphenyl; When "---" is a double bond, R １３ is hydrogen, -CH ２ OH, -CH(OH)-CH ２ OH, -NH ２ , -CH(OH)CH ３ , -OCH ３ , or -NH-(CH ２ ) ２ OH and R １４ does not exist, or When "---" is a single bond, R １３ and R １４ together form =O, R ５ and R ６ Each of is independently hydrogen or -CH ３ (It is.)

4. The TRPC5 inhibitor compound has the structural formula (II): 【Chemistry 4】 3. The method of claim 2 having the formula: R １１ is chloro, -CF ３ , -CHF ２ , or -CH ３ and R １２ is hydrogen or fluoro, R １３ is hydrogen, -NH ２ , -CH ２ OH, or CH(OH)-CH ２ OH).

5. R １１ Ga-CHF ２ and R １２ The method of claim 4 , wherein is fluoro.

6. The method according to claim 3, wherein the TRPC5 inhibitor compound is selected from any one of the following compounds or a pharma- ceutically acceptable salt thereof: 【Table 1-1】 【Table 1-2】 【Table 1-3】 【Table 1-4】

7. The method according to claim 6, wherein the TRPC5 inhibitor compound is selected from any one of the following compounds or a pharma- ceutically acceptable salt thereof: 【Table 2-1】 【Table 2-2】

8. The method according to claim 7, wherein the TRPC5 inhibitor compound is the following compound or a pharma- ceutically acceptable salt thereof: 【Table 3】

9. The method of any one of claims 1 to 8, wherein the immunomodulatory agent is rituximab.

10. 9. The method of any one of claims 1 to 8, wherein the angiotensin converting enzyme inhibitor is captopril, zofenopril, enalapril, ramipril, quinapril, perindopril, lisinopril, benazepril, imidapril, trandolapril, or cilazapril.

11. The method of any one of claims 1 to 8, wherein the angiotensin receptor blocker is losartan, candesartan, valsartan, irbesartan, telmisartan, eprosartan, olmesartan, azilsartan, or fimasartan.

12. The method according to any one of claims 1 to 8, wherein the renin-angiotensin-aldosterone system inhibitor is aliskiren.

13. The method of any one of claims 1 to 8, wherein the endothelin 1 receptor antagonist is ambrisentan, atrasentan, bosentan, or sparsentan.

14. The method of any one of claims 1 to 8, wherein the antiproliferative agent is mycophenolate mofetil.

15. The method according to any one of claims 1 to 8, wherein the SGLT2 inhibitor is canagliflozin, dapagliflozin, empagliflozin, a combination of empagliflozin and linagliptin, a combination of empagliflozin and metformin, or a combination of dapagliflozin and metformin.

16. The method of any one of claims 1 to 8, wherein the calcineurin inhibitor is cyclosporin A or tacrolimus.

17. The method of any one of claims 1 to 8, wherein the nuclear factor-1 (erythroid-derived 2)-like 2 agonist is bardoxolone or CXA-10.

18. The method of any one of claims 1 to 8, wherein the chemokine receptor 2 inhibitor is PF-04136309 or ccx140.

19. The method of any one of claims 1 to 8, wherein the second therapeutic agent is tacrolimus, cyclosporine A, rituximab, mycophenolate mofetil, a corticosteroid, sparsentan, enalapril, or losartan.

20. 20. The method of claim 19, wherein the second therapeutic agent is enalapril, losartan, or cyclosporine A.

21. The disease or condition is selected from the group consisting of focal segmental glomerulosclerosis (FSGS), primary focal segmental glomerulosclerosis, hereditary focal segmental glomerulosclerosis, secondary focal segmental glomerulosclerosis, diabetic nephropathy, Alport syndrome, hypertensive kidney disease, nephrotic syndrome, steroid-resistant nephropathy, minimal change nephrotic syndrome, membranous nephropathy, idiopathic membranous nephropathy, membranoproliferative glomerulonephritis (MPGN), immune complex-mediated MPGN, 21. The method of any one of claims 1 to 20, wherein the glomerulonephritis is selected from the group consisting of pulmonary glomerulonephritis, pulmonary ne ...

22. 22. The method of claim 21, wherein the disease or condition is focal segmental glomerulosclerosis (FSGS), primary focal segmental glomerulosclerosis, hereditary focal segmental glomerulosclerosis, transplant-associated FSGS, or secondary focal segmental glomerulosclerosis.

23. 22. The method of claim 21, wherein the renal disease is a proteinuric renal disease.

24. 22. The method of claim 21, wherein the renal disease is microalbuminuric or macroalbuminuric renal disease.

25. The method of any one of claims 1 to 24, wherein the subject is a human.

26. 26. The method of claim 22 or 25, wherein the disease or condition is focal segmental glomerulosclerosis.

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