Heterocyclic inhibitors of methionine adenosyltransferase 2A.

Abstract

The present invention provides a heterocyclic inhibitor of methionine adenosyltransferase 2A, the structure of which is represented by general formula I. Each substituent is defined as described herein. It also provides a method for its preparation. The compound represented by general formula I has a remarkable inhibitory activity against methionine adenosyltransferase 2A, and is useful for treating methionine adenosyltransferase 2A-mediated diseases. [Formula 1] TIFF2025500974000044.tif44170

Description

[Technical field]

[0001] The present invention belongs to the technical field of medicine, specifically to heterocyclic inhibitors of methionine adenosyltransferase 2A, their preparation methods and uses. [Background technology]

[0002] Methionine adenosyltransferase (MAT), also known as S-adenosylmethionine synthase, is a cellular enzyme that catalyzes the synthesis of S-adenosylmethionine (SAM or AdoMet) from methionine and ATP, and is considered the rate-limiting step of the methionine cycle. SAM is an aminopropyl group donor in polyamine biosynthesis and the main methyl donor used in DNA methylation, and is involved in gene transcription, cell proliferation, and secondary metabolite formation. There are three human isozymes of MAT, including MAT1, MAT2, and MAT3, of which MAT1 and MAT3 are expressed in liver tissue, and methionine adenosine transferase 2A (MAT2A), a subtype of MAT2, is widely expressed in human cell types and is the predominant form in human cancers, and also plays an important role in the production of SAM.

[0003] MTAP (methylthioadenosine phosphorylase) is an enzyme widely expressed in normal tissues that catalyzes the conversion of methylthioadenosine (MTA) to adenine and 5-methylthioribose-1-phosphate, which is converted to adenine monophosphate and 5-methylthioribose-1-phosphate to methionine and formate. MTA is useful as an alternative purine source when purine synthesis is blocked, for example, by antimetabolites.

[0004] The gene encoding MTAP is located at a single site on chromosome 9 and is frequently deleted in cells of the central nervous system, pancreas, esophagus, bladder, and lungs in cancer patients. Loss of MTAP leads to accumulation of MTA compared with MTAP-expressing cells, making MTAP-deficient cells more dependent on SAM production and, therefore, MAT2A activity. In a screen of approximately 400 cancer cell lines, cells lacking MTAP due to MAT2A knockdown lost a greater proportion of vitality compared with cells normally expressing MTAP. Furthermore, inducible knockdown of the MAT2A protein reduces tumor growth in vivo. These results suggest that MAT2A inhibitors may provide a new therapeutic approach for patients with MTAP-deficient tumors.

[0005] Currently, Chinese patent application CN109890822A discloses pyrazolopyrimidinone MAT2A inhibitors, and WO2020123395A1 discloses 2-oxoquinazoline derivatives as MAT2A inhibitors. The present invention provides novel heterocyclic methionine adenosyltransferase 2A inhibitors. Summary of the Invention

[0006] The present invention provides compounds according to Formula I, or a pharma- ceutically acceptable salt thereof. [ka] (However, X is CR 3 or N, and Y is selected from CR 4 or N, Z is selected from CR 5 or N, W is selected from CR 6 or N, Here, R 3 , R 4 , R 5 , and R 6 are each independently hydrogen, cyano, C2-C6 alkynyl, halogen, hydroxy, NH2, (C1-C6 alkyl)-NR 7-, (C1-C6 alkyl)-O-, (C1-C6 alkyl)-S-, C1-C6 alkyl, C3-C6 cycloalkyl, 6-10 membered aryl, C2-C6 alkenyl, or C3-C6 cycloalkenyl, wherein the C1-C6 alkyl, C2-C6 alkenyl, C3-C6 cycloalkyl, or C3-C6 cycloalkenyl, by itself or as part of another group, is selected from halogen, cyano, hydroxy, -NR 7 R 8 , C1-C3 alkyl, C1-C3 alkoxy, C2-C6 alkenyl, or C2-C6 alkynyl, and the 6-10 membered aryl is optionally substituted with halogen, hydroxy, cyano, -NR 7 R 8 , NO2, C1-C3 alkyl, C1-C3 alkoxy, C2-C6 alkenyl, or C2-C6 alkynyl, wherein the C1-C3 alkyl, C1-C3 alkoxy, C2-C6 alkenyl, or C2-C6 alkynyl is optionally substituted with halogen, hydroxy, cyano, -NR 7 R 8 or optionally substituted by NO2, R 1 , and R 2 are each independently selected from a 6- to 10-membered aryl or a 9- to 18-membered benzoheterocyclyl, and the 6- to 10-membered aryl or the 9- to 18-membered benzoheterocyclyl is selected from halogen, hydroxy, cyano, -NR 7 R 8 , NO2, -NR 9 C(O)R 10 , C1-C6 alkyl, (C1-C6 alkyl)-O-, -C(O)NR 9 R 10 or a 5- to 7-membered heteroaryl, wherein the C1-C6 alkyl, by itself or as part of another group or as a 5- to 7-membered heteroaryl, is optionally substituted with halogen, cyano, hydroxy, C1-C3 alkyl, (C1-C3 alkyl)-O-, or -NR 7 R 8 optionally replaced by R 7 , R 8 , R 9 , and R 10are each independently selected from H or C1-C6 alkyl; The condition is that at most two of W, X, Y, and Z are simultaneously N.)

[0007] In some embodiments, X is CR 3 is selected from.

[0008] In some embodiments, Y is CR 4 is selected from.

[0009] In some embodiments, Z is CR 5 is selected from.

[0010] In some embodiments, W is N.

[0011] In some embodiments, R 3 , R 4 , R 5 , and R 6 are each independently hydrogen, halogen, hydroxy, NH2, (C1-C6 alkyl)-NR 7 -, (C1-C6 alkyl)-O-, C1-C6 alkyl, C3-C6 cycloalkyl, or 6-10 membered aryl, where the C1-C6 alkyl, by itself or as part of another group or as a C3-C6 cycloalkyl, is selected from halogen, cyano, hydroxy, or -NR 7 R 8 and said 6-10 membered aryl is optionally substituted by halogen substituted C1-C3 alkoxy.

[0012] In some exemplary embodiments, R 3 , R 4 , R 5 , and R 6 are each independently hydrogen, halogen, hydroxy, NH2, (C1-C6 alkyl)-NR 7-, (C1-C6 alkyl)-O-, C1-C6 alkyl, C3-C6 cycloalkyl, or 6-10 membered aryl, wherein the C1-C6 alkyl is optionally substituted by halogen, either by itself or as part of another group or as C3-C6 cycloalkyl, and the 6-10 membered aryl is optionally substituted by halogen-substituted C1-C3 alkoxy.

[0013] In some exemplary embodiments, R 3 , R 4 , R 5 , and R 6 are each independently hydrogen, halogen, hydroxy, NH2, (C1-C6 alkyl)-NR 7 -, (C1-C6 alkyl)-O-, C1-C6 alkyl, C3-C6 cycloalkyl, or 6-10 membered aryl, wherein the C1-C6 alkyl is optionally substituted by fluorine, either by itself or as part of another group or as C3-C6 cycloalkyl, and the 6-10 membered aryl is optionally substituted by fluorine-substituted C1-C3 alkoxy.

[0014] In some more exemplary embodiments, R 3 is selected from hydrogen, C1-C6 alkyl, or C3-C6 cycloalkyl, preferably R 3 is selected from hydrogen or C1-C6 alkyl.

[0015] In some more exemplary embodiments, R 3 is selected from hydrogen, methyl or cyclopropyl, preferably R 3 is selected from hydrogen or methyl, more preferably R 3 is selected from hydrogen.

[0016] In some more exemplary embodiments, R 4 is selected from hydrogen or C1-C6 alkyl, preferably R 4 is selected from hydrogen or methyl, more preferably R 4 is selected from hydrogen.

[0017] In some more exemplary embodiments, R 5 is hydrogen, halogen, hydroxy, (C1-C6 alkyl)-NR 7 -, (C1-C6 alkyl)-O-, C3-C6 cycloalkyl, or 6-10 membered aryl, wherein the C1-C6 alkyl is optionally substituted by fluorine, either by itself or as part of another group or as C3-C6 cycloalkyl, and the 6-10 membered aryl is optionally substituted by fluorine-substituted C1-C3 alkoxy.

[0018] In some more exemplary embodiments, R 5 is selected from hydrogen, chlorine, hydroxy, cyclopropyl, CF3CH2O-, CHF2O-, CF3CH2NH-, 4-difluoromethoxyphenyl, or CH3CH2O-, preferably R 5 is selected from cyclopropyl, CF3CH2O-, CF3CH2NH-, or CH3CH2O-, more preferably R 5 is selected from cyclopropyl, CF3CH2O-, or CF3CH2NH-, most preferably R 5 is selected from cyclopropyl.

[0019] In some embodiments, R 1 , and R 2 are each independently selected from phenyl, a group represented by the formula: 7 R 8 , NO2, -NR 9 C(O)R 10 , C1-C6 alkyl, (C1-C6 alkyl)-O-, -C(O)NR 9 R 10 or a 5- to 7-membered heteroaryl, wherein the C1-C6 alkyl, by itself or as part of another group or as a 5- to 7-membered heteroaryl, is optionally substituted with halogen, cyano, hydroxy, C1-C3 alkyl, (C1-C3 alkyl)-O-, or -NR 7 R 8 is optionally replaced by [ka]

[0020] In some embodiments, R 1 , and R 2 are each independently selected from phenyl, a group represented by the formula: [ka]

[0021] In some embodiments, R 7 , R 8 , R 9 , and R 10 are each independently selected from H.

[0022] In some embodiments, R 1 is selected from phenyl, a group represented by the formula: wherein said group is optionally substituted by C1-C6 alkyl, or (C1-C6 alkyl)-O-, said C1-C6 alkyl being optionally substituted by halogen, either by itself or as part of another group. [ka]

[0023] In some embodiments, R 1 is selected from the groups represented by the formula: [ka] Preferably, R 1 is selected from the groups represented by the following formulas: [ka]

[0024] In some embodiments, R 2 is selected from phenyl or a group represented by the following formula, wherein said group is optionally substituted with (C1-C6 alkyl)-O-, and said C1-C6 alkyl is optionally substituted with halogen. [ka]

[0025] In some embodiments, R 2 is selected from the groups represented by the formula: [ka] Preferably, R 2 is selected from the groups represented by the following formulas: [ka]

[0026] In some embodiments, the compound of Formula I has the structure shown in Formula II: [ka] (However, R 1 , R 2 , R 3 , R 4 , and R 5 The definitions of are the same as those in the compound of formula I.

[0027] In another aspect, the present invention provides the following compound or a pharma- ceutically acceptable salt thereof: [ka]

[0028] In some embodiments, the present invention provides a pharmaceutical composition comprising a therapeutically effective amount of the aforementioned compound or a pharma- ceutically acceptable salt thereof.

[0029] In some embodiments, the present invention provides a pharmaceutical composition comprising a therapeutically effective amount of the aforementioned compound, or a pharma- ceutically acceptable salt thereof, and a pharma- ceutically acceptable carrier.

[0030] The pharmaceutical compositions of the present invention may be administered by any suitable route or method, such as orally or parenterally (e.g., intravenously). The therapeutically effective amount of the compound is about 1 mg to 1 g per kg body weight per day.

[0031] For oral administration, the pharmaceutical composition of the present invention is usually provided in the form of a tablet, capsule or solution. The tablet may contain the compound of the present invention or a pharma- ceutically acceptable salt thereof and a pharma- ceutically acceptable carrier. The carrier includes, but is not limited to, a diluent, a disintegrant, a binder, and a lubricant.

[0032] For parenteral administration, the pharmaceutical composition of the present invention may be administered by intravenous injection, intramuscular injection or subcutaneous injection. It is usually provided as a sterile aqueous solution or suspension or lyophilized powder, with appropriate adjustment of pH and isotonicity.

[0033] In another aspect, the present invention also provides the use of the aforementioned compound or a pharma- ceutically acceptable salt thereof or a pharmaceutical composition thereof in the manufacture of a medicament for the prophylaxis and / or treatment of a MAT2A mediated disease or condition.

[0034] In another aspect, the present invention also provides a method for preventing and / or treating a MAT2A-mediated disease or condition, comprising administering to an individual in need thereof an effective amount of the aforementioned compound, a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition thereof.

[0035] In another aspect, the present invention also provides the above-mentioned compound of the present invention, its pharma- ceutically acceptable salt or the pharmaceutical composition of the present invention for preventing and / or treating a MAT2A-mediated disease or condition, such as colorectal cancer.

[0036] In some specific embodiments, said MAT2A mediated disease is a MAT2A overexpression mediated disease.

[0037] In another aspect, the present invention provides methods for preparing compounds of Formula I or II, including, but not limited to, the following synthetic schemes. [ka] (wherein X is selected from chlorine, bromine, or iodine; R 1 , R 2 , and R 5 is defined as in formula I above, and R a , and R b are each independently selected from C1 to C3 alkyl.

[0038] In some embodiments, R a , and R b are each independently selected from methyl.

[0039] The compound of formula 1-1 is reacted with the compound of formula 1-2 to produce the compound of formula 1-3, the compound of formula 1-3 is reacted with hydrohalic acid to produce the compound of formula 1-4, the compound of formula 1-4 is subjected to a halogenation reaction to produce the compound of formula 1-5, the compound of formula 1-5 is reacted with the compound of formula 1-6 to produce the compound of formula 1-7, the compound of formula 1-7 is subjected to a cyclization reaction to produce the compound of formula 1-8, the compound of formula 1-8 is reacted with the compound of formula 1-9a or 1-9b to produce the compound of formula 1-10, the compound of formula 1-10 is reacted with the compound of formula 1-11a or 1-11b to produce the compound of formula 1-12, or the compound of formula 1-8 is reacted with the compound of formula 1-9a or 1-9b to produce the compound of formula 1-13, in which in the compound of formula 1-13, R 1 , and R 2 is the same.

[0040] In some embodiments, X is selected from chlorine.

[0041] In some embodiments, R 1 , and R 2are each independently selected from the groups represented by the formula: 5 is selected from cyclopropyl. [ka] [Brief description of the drawings]

[0042] [Figure 1] 1 shows a tumor inhibition curve of HCT116 MTAP- / - tumor transplant model in Test Example 3. [Diagram 2] 13 shows the results of SAM inhibition within the tumor in the HCT116 MTAP- / - tumor transplant model in Test Example 3. [Diagram 3] 1 shows a tumor inhibition curve of a KP-4 tumor transplant model in Test Example 4. [Figure 4] 13 shows the results of SAM inhibition within the tumor in the KP-4 tumor transplant model in Test Example 4. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0043] definition Unless otherwise stated, the following terms used in the specification and claims have the following meanings.

[0044] The terms "optional" or "optionally" mean that the subsequently described event or circumstance may or may not occur, and the description includes cases where the event or circumstance occurs and cases where it does not occur.

[0045] Numerical ranges in this specification refer to individual integers within the given range. For example, "C1-C6" means that the group may have 1 carbon atom, 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, 5 carbon atoms, or 6 carbon atoms. "C3-C6" means that the group may have 3 carbon atoms, 4 carbon atoms, 5 carbon atoms, or 6 carbon atoms.

[0046] The term "membered" refers to the number of skeletal atoms or atomic groups constituting the ring. For example, "5- to 7-membered" means that the number of skeletal atoms or atomic groups constituting the ring is 5, 6, or 7. Thus, for example, pyridine, piperidine, piperazine, and benzene are 6-membered rings, and thiophene and pyrrole are 5-membered rings.

[0047] The term "substituted" means that any one or more of the hydrogen atoms on a particular group are replaced with a substituent, as long as the valence of the particular group is normal and the substituted compound is stable. For example, "substituted with halogen" means that any one or more of the hydrogen atoms on a particular group are replaced with a halogen, as long as the valence of the particular group is normal and the substituted compound is stable.

[0048] The term "alkyl" refers to a saturated aliphatic hydrocarbon group, including straight-chain or branched-chain saturated hydrocarbon groups having the indicated number of carbon atoms. For example, the term "C1-C6 alkyl" includes C1 alkyl, C2 alkyl, C3 alkyl, C4 alkyl, C5 alkyl, or C6 alkyl, examples of which include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, t-butyl, n-pentyl, 2-pentyl, 3-pentyl, n-hexyl, 2-hexyl, or 3-hexyl, etc.

[0049] The term "cycloalkyl" refers to a monocyclic saturated hydrocarbon system with no heteroatoms or double bonds. Examples of the term "3- to 6-membered cycloalkyl" include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl.

[0050] The term "halogen" refers to fluorine, chlorine, bromine and iodine.

[0051] The term "aryl" refers to an all-carbon monocyclic or fused bicyclic aromatic ring group having a conjugated pi-electron system obtained by removing a hydrogen atom from a single carbon atom of a parent aromatic ring system. It includes bicyclic groups fused to a saturated ring, a partially unsaturated ring, or an aromatic carbocyclic ring. Examples include, but are not limited to, phenyl, naphthyl, anthracenyl, indene, indane, 1,2-dihydronaphthalene, or 1,2,3,4-tetrahydronaphthalene.

[0052] The term "heteroaryl" refers to a monovalent aryl containing at least one heteroatom independently selected from nitrogen, oxygen, and sulfur. For example, examples of "5- to 7-membered heteroaryl" include, but are not limited to, pyridyl, thienyl, imidazolyl, pyrimidinyl, pyridyl, furyl, pyrazinyl, or thiazolyl.

[0053] The term "9-18 membered benzoheterocyclyl" refers to a ring system having 9-18 ring atoms or ring atom groups formed by the condensation of a benzene ring and a heterocyclic ring, the benzene ring and the heterocyclic ring sharing one pair of adjacent ring atoms, and the linking site to the core structure is located in the benzene ring portion. The heterocyclic portion is a 5-12 membered saturated, partially unsaturated or fully unsaturated ring system having ring carbon atoms and 1-4 cycloheteroatoms or heteroatom groups, the heteroatoms or heteroatom groups being independently selected from nitrogen, sulfur, oxygen, sulfoxide, sulfone, and the groups shown in the formula: [ka] Heterocycles may be monocyclic, bicyclic, or tricyclic systems, where two or more rings are present in the form of fused, spiro, or bridged rings, examples of which include, but are not limited to, the groups shown in the formula: [ka]

[0054] Of the structure shown below, [ka] The structures shown below indicate the points of attachment of chemical bonds. [ka] When the following structure appears in a bicyclic or polycyclic ring and the linking position is indefinite, as far as valence allows, [ka] The linking site is meant to be limited to any atom on the single ring in which the structure shown below resides. [ka] For example, the group shown in the formula below means that the linking site is at any carbon atom located only on the benzene ring in the bicycle, satisfying the valence bond requirements. [ka]

[0055] The term "pharmaceutically acceptable salts" refers to salts that retain the biological effectiveness of the free acids and bases of a particular compound without adverse biological effects, such as acid (including organic and inorganic acids) addition salts and base addition salts (including organic and inorganic bases).

[0056] The pharma- ceutically acceptable salts of the present invention can be synthesized from parent compounds containing acids or bases by conventional chemical methods. In general, the preparation of such salts is carried out by reacting the free acid or base form of these compounds with a stoichiometrically appropriate base or acid in water or an organic solvent, or in a mixture of both.

[0057] The term "effective amount" or "therapeutically effective amount" refers to a non-toxic but sufficient amount of a drug or agent to achieve a desired effect.

[0058] The term "pharmaceutical acceptable carrier" refers to a carrier that has no obvious irritating effect on the living body and does not impair the biological activity and performance of the active compound, including, but not limited to, diluents, disintegrants, binders, flow aids, and wetting agents that are approved by the State Food and Drug Administration and can be used by humans or animals.

[0059] The abbreviations used in the claims and specification have the following meanings: M:mol / L mM:mmol / L μM: μmol / L nM:nmol / L LCMS: Liquid Chromatography-Mass Spectrometry Brij35: Laureth polyoxyethylene ether BSA: bovine serum albumin DMSO: dimethyl sulfoxide; rpm: revolutions per minute Tris-HCl: Trishydroxymethylaminomethane hydrochloride O.D. 620 : Absorbance at wavelength 620 nm SAM: S-adenosylmethionine

[0060] The production methods of the compounds of the present invention will be described in more detail below, but these specific production methods are not intended to limit the scope of the present invention. Furthermore, reaction conditions such as reactants, solvents, bases, amounts of compounds used, reaction temperatures, and reaction times are not limited to the following examples.

[0061] The compounds of the present invention can also be readily produced by any combination of various synthetic methods described herein or known in the art, and such combinations can be readily performed by one of ordinary skill in the art.

[0062] Working Example Example 1: 2-Cyclopropyl-7,9-bis[4-(difluoromethoxy)phenyl]-8H-pyrimido[1,2-b]pyridazin-8-one [ka]

[0063] a) Preparation of N-(5-methoxypyridazin-3-yl)acetamide 3-Chloro-5-methoxypyridazine (1g), acetamide (0.61g), tris(dibenzylideneacetone)dipalladium(0) (0.32g), 4,5-bis(diphenylphosphine)-9,9-dimethylxanthene (0.40g), cesium carbonate (6.76g) were added to the reaction flask in sequence, followed by 1,4-dioxane (100mL), stirred under nitrogen protection at 100°C for 3 hours, concentrated under reduced pressure, evaporated to dryness to obtain a residue, extracted with ethyl acetate (3x50mL), combined organic phase washed with saturated brine (1x50mL), dried over anhydrous sodium sulfate, filtered, the filtrate concentrated under reduced pressure, purified by silica gel column chromatography (mobile phase: ethyl acetate / petroleum ether=10 / 1(V / V)) to obtain 780mg of the title compound. LCMS m / z= 168.05[M+1] +

[0064] b) Preparation of 6-aminopyridazin-4(1H)-one N-(5-methoxypyridazin-3-yl)acetamide (500 mg) and an aqueous hydrogen bromide solution (15 mL) were added to a microwave tube, and the mixture was stirred at 140° C. for 5 hours under microwave irradiation and concentrated under reduced pressure to give 360 ​​mg of the title compound. LCMS m / z=111.95[M+1] +

[0065] c) Preparation of 6-amino-3,5-dibromopyridazin-4(1H)-one A reaction flask was charged with 6-aminopyridazin-4(1H)-one (360 mg), N-bromosuccinimide (1.73 g), stirred in N,N-dimethylformamide (4 mL) at room temperature for 2 hours, concentrated under reduced pressure and evaporated to dryness to give a residue, which was washed with acetonitrile (3×5 mL) to give 360 ​​mg of the title compound.

[0066] d) Preparation of 6-amino-3,5-dibromo-1-[(1E)-3-cyclopropyl-3-oxoprop-1-en-1-yl]pyridazin-4(1H)-one 6-Amino-3,5-dibromopyridazin-4(1H)-one (430 mg), (2E)-3-chloro-1-cyclopropylprop-2-en-1-one (167.0 mg), and potassium carbonate (663.0 mg) were added to a reaction flask, and the mixture was stirred overnight at 25°C in N,N-dimethylformamide (43 mL) and concentrated under reduced pressure to obtain a mixture. The pH of the mixture was adjusted to 2-3 with 1M hydrochloric acid, and the mixture was washed with water (3 x 10 mL) to obtain 335 mg of the title compound.

[0067] e) Preparation of 7,9-dibromo-2-cyclopropyl-8H-pyrimido[1,2-b]pyridazin-8-one 6-amino-3,5-dibromo-1-[(1E)-3-cyclopropyl-3-oxoprop-1-en-1-yl]pyridazin-4(1H)-one (355 mg) was added to the reaction flask, followed by a solution of 2 mol / L hydrogen chloride in 1,4-dioxane (10 mL), stirred at 25° C. for 1 hour, concentrated under reduced pressure to obtain a dry residue, washed with aqueous sodium bicarbonate (1×10 mL), filtered to collect the filter cake, and washed with water (3×5 mL) to obtain 320 mg of the title compound.

[0068] f) Preparation of 2-cyclopropyl-7,9-bis[4-(difluoromethoxy)phenyl]-8H-pyrimido[1,2-b]pyridazin-8-one 7,9-Dibromo-2-cyclopropyl-8H-pyrimido[1,2-b]pyridazin-8-one (60 mg), 4-(difluoromethoxy)phenylboronic acid (98.1 mg), water (0.4 mL), 1,4-dioxane (2 mL), potassium phosphate (184.6 mg), and [1,1'-bis(diphenylphosphino)ferrocene]palladium(II) dichloride dichloromethane complex (14.17 mg) were added to a reaction flask, and the mixture was stirred at 80°C under nitrogen protection for 1 hour, concentrated under reduced pressure to obtain a dry residue, which was separated to obtain 37 mg of the title compound. Preparative conditions: Chromatography column: XBridge PrepOBD C18 column, 30*150mm, 5μm; Column temperature: 25℃; Mobile phase A: water (10mmol / LNH4HCO3), Mobile phase B: acetonitrile; Flow rate: 60mL / min; Elution gradient: 45%B-85%B elution from 0 to 10min, 85%B elution from 10min onwards; Detection wavelength: UV220nm; Retention time (min): 7.32. 1 H NMR(400MHz,DMSO-d6)δ8.72(d,J= 7.2Hz,1H),8.27-8.21(m,2H),7.64-7.58(m,2H),7.56-7.26(m,3H),7.21-7 .11(m,3H),7.07(d,J=7.2Hz,1H),2.20(tt,J=8.1,4.5Hz,1H),1.18-1.10(m, 2H),1.01(p,J=3.8Hz,2H). LCMS m / z=472[M+H] +

[0069] Example 2: 7,9-Bis(benzo[d][1,3]dioxo-5-yl)-2-cyclopropyl-8H-pyrimido[1,2-b]pyridazin-8-one [ka]

[0070] This was prepared by following the preparation method of Example 1, except that in step f) 4-(difluoromethoxy)phenylboronic acid was changed to benzo[d][1,3]dioxo-5-ylboronic acid (the group shown in the formula below). [ka] 1 H NMR(400MHz,DMSO-d6)δ8.69(d,J=7.2Hz,1H),7.88(dd,J=8.3,1.7Hz,1H),7.76(d,J=1.7Hz,1H),7.09-6.99(m,4H),6.93( d,J=8.0Hz,1H),6.11(s,2H),6.04(s,2H),2.18(td,J=8.0,4.1Hz,1H),1.12(dt,J=6.7,3.4Hz,2H),1.01(t,J=3.8Hz,2H). LCMS m / z=428[M+1] +

[0071] Example 3: 2-Cyclopropyl-9-[4-(difluoromethoxy)phenyl]-7-(2-methyl-2H-indazol-5-yl)-8H-pyrimido[1,2-b]pyridazin-8-one [ka]

[0072] a) Preparation of 9-bromo-2-cyclopropyl-7-(2-methyl-2H-indazol-5-yl)-8H-pyrimido[1,2-b]pyridazin-8-one 7,9-Dibromo-2-cyclopropyl-8H-pyrimido[1,2-b]pyridazin-8-one (50 mg), 2-methylindazol-5-ylboronic acid (28.06 mg), potassium carbonate (60.09 mg), [1,1'-bis(diphenylphosphino)ferrocene]palladium(II) dichloride dichloromethane complex (11.81 mg), 1,4-dioxane (0.5 mL), and water (0.1 mL) were added to the reaction flask in sequence, and the mixture was stirred at 60°C for 1 hour under nitrogen protection. The reaction mixture was concentrated under reduced pressure and purified by silica gel column chromatography (mobile phase: ethyl acetate / petroleum ether = 4 / 1 (V / V)) to obtain 40 mg of the title compound. 1H NMR(400MHz,DMSO)δ8.98(s,1H),8.81(d,J=7.1Hz,1H),8.54(s,1H),7.95 (d,J=8.9Hz,1H),7.66(d,J=9.1Hz,1H),7.14(d,J=7.1Hz,1H),4.20(s,3H),2.34(s,1H),1.46-1.10(m,4H). LCMS m / z=396[M+1] +

[0073] b) Preparation of 2-cyclopropyl-9-[4-(difluoromethoxy)phenyl]-7-(2-methyl-2H-indazol-5-yl)-8H-pyrimido[1,2-b]pyridazin-8-one 9-Bromo-2-cyclopropyl-7-(2-methyl-2H-indazol-5-yl)-8H-pyrimido[1,2-b]pyridazin-8-one (35 mg), 4-(difluoromethoxy)phenylboronic acid (24.90 mg), potassium carbonate (36.62 mg), [1,1'-bis(diphenylphosphino)ferrocene]palladium(II) dichloride dichloromethane complex (7.20 mg), 1,4-dioxane (1 mL), and water (0.2 mL) were added to the reaction flask in sequence, and the mixture was stirred at 80°C for 1 hour under nitrogen protection. The reaction mixture was concentrated under reduced pressure and separated to give the title compound (14.4 mg). Preparative conditions: Chromatography column: XBridge PrepOBD C18 column, 30*150mm, 5μm; Column temperature: 25℃; Mobile phase A: water (10mmol / LNH4HCO3), Mobile phase B: acetonitrile; Flow rate: 60mL / min; Elution gradient: 25%B-70%B elution from 0 to 10min, 85%B elution from 10min onwards; Detection wavelength: UV220nm; Retention time (min): 6.8. 1H NMR(400MHz,DMSO-d6)8.97(s,1H),8.76(d,J=7.2Hz,1H),8.52(s,1H),7.97(dd,J=9.2,1.6Hz,1H),7.68-7.59(m,3H),7.31(t,J=74.4Hz,1H ),7.22-7.16(m,2H),7.06(d,J=7.2Hz,1H),4.20(s,3H),2.19(dq,J=8.1,4.6,4.0Hz,1H),1.13(dd,J=7.8,3.5Hz,2H),1.02(t,J=3.8Hz,2H). LCMS m / z=460[M+1] +

[0074] Test example 1: Biological activity test 1. MAT2A Method 1. Experimental Steps a) First, 5x MAT2A test buffer (250 mM Tris-HCl, pH 8.0; 250 mM KCl; 75 mM MgCl2; 0.025% BSA; 0.05% Brij35; 1.5 mM EDTA) was prepared and partially diluted to 1x in preparation for use.

[0075] b) Preparation and addition of MAT2A enzyme (BPS, 71401) MAT2A enzyme was adjusted to 3.674 ng / μL (1.67×, final concentration 2.20 ng / μL) in 1× MAT2A test buffer, and 15 μL of the 1.67× MAT2A enzyme solution was added to each compound test well and negative control well, and 15 μL of 1× MAT2A test buffer was added to blank control wells using a BioTek (MultiFlo FX) automated separatory counter.

[0076] c) Preparation and addition of compounds The test compounds were diluted to 100μM from a 10mM stock solution using DMSO, and the positive drug AGI-24512 was diluted under the same conditions, and then automatically sprayed into each well according to the set concentration gradient using a Tecan compound titrator (D300e), with the volume sprayed being so small that it can be ignored. The concentration gradient started at 1μM, with 1 / 2 log dilution, and a total of eight gradients were set. The mixture was centrifuged at 2500rpm for 30 seconds and incubated at 25℃ for 30 minutes.

[0077] d) Preparation of ATP 10 mM ATP (Sigma, A7699) was diluted to 700 μM using 1× MAT2A test buffer and was ready for use.

[0078] e) Preparation and addition of substrate and ATP mixture 5x MAT2A test buffer (3μL / well), 750μM L-methionine (Adamas, 01100469) (2.5μL / well), 700μM ATP (2.5μL / well), double distilled water (2μL / well). The total amount of the mixture required was prepared according to the number of detection wells, and 10μL was added per well using a BioTek (MultiFlo FX) automatic dispenser. The mixture was centrifuged at 2500rpm for 30 seconds and reacted at 25℃ for 150 minutes.

[0079] f) Addition of Biomol Green detection reagent 50 μL of Biomol Green (Enzo, BML-AK111) was added per well using a BioTek (MultiFlo FX) automated dispenser, centrifuged at 2500 rpm for 30 seconds, and incubated at 25°C for 20 minutes. g) After the reaction is complete, the OD is measured using a Perkin Elmer (Envision 2105) multifunction plate reader. 620 It was read as follows.

[0080] 2. Data Analysis The formula for calculating the inhibition rate is as follows:

number

[0081] however, OD sample: OD of sample well 620 value ODmin: OD of blank control wells containing neither enzyme nor test compound 620 Represents the average value. ODmax: OD of negative control wells with enzyme but no compound 620 Represents the average value. Dose-response curves were fitted using GraphPad Prism 5 software log(inhibitor) vs. response-variable slope to obtain IC values ​​for compound-induced inhibition of MAT2A enzyme. 50 Got the value.

[0082] II. Cell test method 1. Experimental Steps HCT116 MTAP- / - cells (purchased from Horizon Discovery): MTAP gene-deficient human colorectal cancer cell line was cultured in RPMI 1640 medium + 10% fetal bovine serum (FBS). On day 0 of the experiment, the viable cell density of the above cells in logarithmic growth phase was adjusted to 5000 cells / mL and inoculated into a 96-well plate at 100 μL / well, with a blank group set up in parallel. The inoculated cell plate was placed at 37°C and cultured overnight in a 5% CO2 incubator.

[0083] On the first day of the experiment, the cell plate that had been cultured overnight was removed, the supernatant was discarded, 80 μL of serum-free RPMI 1640 medium was added per well, and the plate was placed in an incubator for 4 hours of starvation culture. The test compound was dissolved in dimethyl sulfoxide (DMSO) to prepare a 10 mM compound mother solution. After starvation was completed, the cell plate was removed and 80 μL of RPMI 1640 + 20% FBS medium was added per well. The cell plate was placed in an automatic medium dispenser D300e (Tecan) and the administration program was set as follows. Compounds were tested up to a maximum concentration of 30 μM, and a 3-fold concentration gradient dilution was performed using DMSO to set a total of 10 concentrations, with two duplicate wells installed for each concentration, and the final DMSO concentration in each well of the 96-well plate was 0.3% (v / v). The previously prepared 10 mM test compound mother solution was removed and administered by running the administration program above. After administration was completed, the cell plate was placed in an incubator and cultured for 120 hours.

[0084] On day 6 of the experiment, the cell plates were removed, 50 μL of CellTiter-Glo® (purchased from Promega) was added per well, and the fluorescent signal was measured on Envision (PerkinElmer) according to the manufacturer's instructions.

[0085] 2. Data Analysis The IC values ​​for the inhibition of cell proliferation by compounds were calculated by fitting the dose-response curves: log(inhibitor) vs. response-variable slope using GraphPad Prism 5 software. 50 The inhibition rate was calculated as follows:

number

[0086] however, Signal value of the test object: Average value of the fluorescence signals of the cells + medium + compound group Signal value of blank group: Mean value of fluorescence signal of medium group Signal value of negative control group: Average value of fluorescent signals of cells + medium group

[0087] Third, experimental results: As a result of the above experimental method, the inhibitory IC value of AGI-24512 against MAT2A was 50 was 26.79 nM. Structure of AGI-24512: [ka]

[0088] The experimental results for the compounds of the present invention are shown in Table 1. [Table 1]

[0089] Test example 2: In vivo pharmacokinetic study in ICR mice 1. Experimental Steps Male ICR mice (6-10 weeks old, Beijing Vital River Laboratory Animal Technology Co., Ltd.) were kept in an SPF animal breeding room with a temperature of 20-25°C, relative humidity of 40%-70%, and 12 hours of light and dark. The animals had free access to water and food. After being kept normally for at least 5 days, the mice were examined by a veterinarian and those with good symptoms were selected for this experiment. Each mouse was numbered on the tail.

[0090] The day before the experiment, the mice were fasted overnight, but allowed to drink water ad libitum and were fed 4 hours after administration. Compounds were prepared into 20 mg / mL stock solutions using DMSO, and appropriate volumes of the 20 mg / mL stock solutions were accurately pipetted into glass bottles, and appropriate volumes of PEG400 were added and mixed, followed by the addition of propylene glycol (PG). The ratio of the solvents in the final formulation was DMSO:PEG400:PG (v / v / v) = 5:65:30, and a test solution for administration of each test compound at a concentration of 1 mg / mL was obtained.

[0091] After weighing the mice, the theoretical administration volume for each mouse was calculated by the following formula: The actual administration volume and blood sample collection time for each mouse should be recorded in detail in the corresponding table.

number

[0092] On the day of the experiment, each group of mice was orally administered with the administration test solution of each test compound at a dose of 10 mg / kg. After administration, approximately 40 μL of blood was collected from the mouse's orbit at each time point and placed in an EDTA-K2 anticoagulant tube. The whole blood sample was centrifuged at 1500 to 1600 g for 10 min, and the separated plasma was stored in a refrigerator at -40 to -20 °C and used for biological sample analysis.

[0093] 2. Data Analysis The concentrations of the compounds in the biological samples obtained in the experiments were measured by LC-MS / MS analysis. The pharmacokinetic parameters were calculated using the non-compartmental model of Pharsight Phoenix 7.0.

[0094] 3. Experimental Results The calculated pharmacokinetic parameters are shown in Table 2. [Table 2]

[0095] Test Example 3: HCT116 MTAP - / - In vivo drug efficacy study in subcutaneous xenograft tumors in nude mice 1. Experimental Steps Female Nu / Nu nude mice (6-8 weeks old, Beijing Vital River Laboratory Animal Technology Co., Ltd.) were housed in an SPF animal breeding room with a temperature of 20-25°C, relative humidity of 40%-70%, and 12 hours of light and dark. The animals had free access to water and food. The animals were housed in an acclimated condition before the experiment.

[0096] HCT116 MTAP - / -Horizon cells were grown in vitro, and cells in the logarithmic growth phase were resuspended in serum-free RPMI-1640 medium at a cell concentration of 6.0 × 10 7 The cell suspension was adjusted to 100 cells / mL. 100 μL of the cell suspension was injected subcutaneously into the anterior right axilla of nude mice using a 1 mL syringe, and the condition of the animals was periodically observed to monitor the growth of the transplanted tumor.

[0097] Tumor volume is 100-300mm 3 When the tumor volume reached 100 mg / kg, animals with tumor volumes too large, too small, or uncertain tumor formation were eliminated, and tumor-bearing mice in good health and with similar tumor volumes were selected and grouped using a random block method. The AG-270 group and the test compound group were orally administered daily (AG-270: 50 mg / kg, structure is as follows; test compound: 5 mg / kg), and the control group was orally administered the same volume of blank solvent every day. During the administration period, the tumor diameter was measured twice a week, the tumor volume was calculated, and the animal's weight was measured and recorded.

[0098] Detection of SAMs in transplanted tumors At the end of the study, the animals were euthanized using CO2, and the tumor tissue was scraped off, washed with cold PBS, weighed, flash frozen in liquid nitrogen, and then cryogenically frozen and stored (-80°C) for use. The frozen tumor tissue was removed and thawed in an ice bath, and then homogenized in 80% aqueous methanol (containing 1M formic acid) at a ratio of 1:10 (w / v) between the tumor tissue and aqueous methanol (containing 1M formic acid). After homogenization, the homogenate was collected and processed to detect S-adenosylmethionine (SAM) using LC-MS / MS.

[0099] AG-270 Structure: [ka]

[0100] 2. Data Analysis The formula for calculating tumor volume (TV) is TV=1 / 2×a×b.2 where a represents the long diameter of the tumor and b represents the short diameter of the tumor.

[0101] The formula for calculating relative tumor volume (RTV) is RTV=TV. t / TV initial where TV initial is the tumor volume measured when administered in each group, and TV t is the tumor volume at each measurement during the treatment period.

[0102] The formula for calculating the relative tumor growth rate (T / C(%)) is T / C%=(RTV T / RTV C ) × 100%, where RTV T indicates the relative tumor volume of the treatment group, RTV C indicates the relative tumor volume in the vehicle control group.

[0103] The formula for tumor growth inhibition (TGI(%)) is TGI=[1-(TV) t(T) -TV initial(T) ) / (TV t(C) -TV initial(C) )] × 100%, where TV t(T) indicates tumor volume per measurement in the treatment group, and TV initial(T) indicates the tumor volume of the treatment group when administered by group, and TV t(C) indicates tumor volume per measurement in the vehicle control group, and TV initial(C) indicates the tumor volume in the solvent control group when administered by group.

[0104] The formula for calculating the animal weight loss rate is Animal Weight Loss Rate = 100% × (BW initial -B.W. t ) / BW initial where BW t indicates the animal body weight at each measurement during the administration period, and BW initial indicates the animal body weight when administered by group.

[0105] The formula for calculating the tumor inhibition rate (IR) based on tumor weight is IR = 100% × (W C -W T ) / W C where W C indicates the tumor weight in the control group, and W T indicates tumor weight of treatment group.

[0106] The test data were calculated and related statistical processing was performed using Microsoft Office Excel 2007 software. Data are expressed as mean ± standard error (Mean ± SE) unless otherwise stated, and t-test was used for comparison between both groups.

[0107] 3. Experimental Results HCT116 MTAP - / - The tumor inhibition curves and intratumoral SAM inhibition results of the transplanted tumor model are shown in Figures 1 and 2, respectively.

[0108] Test Example 4: In vivo drug efficacy test on KP-4 mouse subcutaneous xenograft tumor 1. Experimental Steps Female NOD SCID mice (6-8 weeks old, Beijing Vital River Laboratory Animal Technology Co., Ltd.) were housed in a SPF animal breeding room with a temperature of 20-25°C, relative humidity of 40%-70%, and a 12-hour dark and 12-hour light cycle. The animals had free access to water and food. The animals were housed in an acclimated condition before the study.

[0109] KP-4 cells (Cobioer Biosciences Co., Ltd.) were cultured in vitro and grown. Logarithmic growth phase cells were resuspended in serum-free RPMI-1640 medium to a cell concentration of 1.0 × 10 8 The cell suspension was adjusted to 100 cells / mL. 100 μL of the cell suspension was injected subcutaneously into the anterior right axilla of nude mice using a 1 mL syringe, and the condition of the animals was periodically observed to monitor the growth of the transplanted tumor.

[0110] Tumor volume 80-100mm 3After reaching the age of 10 days, animals with tumor volumes that were too large, too small, or with uncertain tumor formation were eliminated, and tumor-bearing mice in good health and with similar tumor volumes were selected and grouped using a random block method. The AG-270 group and the test compound group were orally administered daily (AG-270: 100 mg / kg, test compound: 2.5 mg / kg), and the control group was orally administered the same volume of blank solvent every day. During the administration period, the tumor diameter was measured twice a week, the tumor volume was calculated, and the animal's weight was weighed and recorded.

[0111] Detection of SAMs in transplanted tumors At the end of the study, the animals were euthanized using CO2, and the tumor tissue was scraped off, washed with cold PBS, weighed, flash frozen in liquid nitrogen, and then cryogenically frozen and stored (-80°C) for use. The frozen tumor tissue was removed and thawed in an ice bath, and then homogenized in 80% aqueous methanol (containing 1M formic acid) at a ratio of 1:10 (w / v) between the tumor tissue and aqueous methanol (containing 1M formic acid). After homogenization, the homogenate was collected and processed to detect S-adenosylmethionine (SAM) using LC-MS / MS.

[0112] 2. Data Analysis The formula for calculating tumor volume (TV) is TV=1 / 2×a×b. 2 where a represents the long diameter of the tumor and b represents the short diameter of the tumor.

[0113] The formula for calculating relative tumor volume (RTV) is RTV=TV. t / TV initial where TV initial is the tumor volume measured when administered in each group, and TV t is the tumor volume at each measurement during the treatment period.

[0114] The formula for calculating the relative tumor growth rate (T / C(%)) is T / C%=(RTV T / RTV C ) × 100%, where RTVT indicates the relative tumor volume of the treatment group, RTV C indicates the relative tumor volume in the vehicle control group.

[0115] The formula for tumor growth inhibition (TGI(%)) is TGI=[1-(TV) t(T) -TV initial(T) ) / (TV t(C) -TV initial(C) )] × 100%, where TV t(T) indicates tumor volume per measurement in the treatment group, and TV initial(T) indicates the tumor volume of the treatment group when administered by group, and TV t(C) indicates tumor volume per measurement in the vehicle control group, and TV initial(C) indicates the tumor volume in the solvent control group when administered by group.

[0116] The formula for calculating the animal weight loss rate is Animal Weight Loss Rate = 100% × (BW initial -B.W. t ) / BW initial where BW t indicates the animal body weight at each measurement during the administration period, and BW initial indicates the animal body weight when administered by group.

[0117] The formula for calculating the tumor inhibition rate (IR) based on tumor weight is IR = 100% × (W C -W T ) / W C where W C indicates the tumor weight in the control group, and W T indicates tumor weight of treatment group.

[0118] The test data were calculated and related statistical processing was performed using Microsoft Office Excel 2007 software. Data are expressed as mean ± standard error (Mean ± SE) unless otherwise stated, and t-test was used for comparison between both groups.

[0119] 3. Experimental Results The tumor inhibition curve and intratumoral SAM inhibition results of the KP-4 tumor xenograft model are shown in Figures 3 and 4, respectively.

Claims

1. A compound of formula I or a pharma- ceutically acceptable salt thereof. 【Chemistry 1】 (however, X is CR ３ or N, and Y is selected from CR ４ or N, Z is selected from CR ５ or N, W is selected from CR ６ or N, Here, R ３ , R ４ , R ５ , and R ６ are each independently hydrogen, cyano, C2-C6 alkynyl, halogen, hydroxy, NH ２ , (C1-C6 alkyl)-NR ７ -, (C1-C6 alkyl)-O-, (C1-C6 alkyl)-S-, C1-C6 alkyl, C3-C6 cycloalkyl, 6- to 10-membered aryl, C2-C6 alkenyl, or C3-C6 cycloalkenyl, wherein the C1-C6 alkyl, C2-C6 alkenyl, C3-C6 cycloalkyl, or C3-C6 cycloalkenyl, by itself or as part of another group, is selected from halogen, cyano, hydroxy, -NR ７ R ８ , C1-C3 alkyl, C1-C3 alkoxy, C2-C6 alkenyl, or C2-C6 alkynyl, wherein the 6-10 membered aryl is optionally substituted with halogen, hydroxy, cyano, -NR ７ R ８ , NO ２ , optionally substituted by C1-C3 alkyl, C1-C3 alkoxy, C2-C6 alkenyl, or C2-C6 alkynyl, wherein said C1-C3 alkyl, C1-C3 alkoxy, C2-C6 alkenyl, or C2-C6 alkynyl is halogen, hydroxy, cyano, -NR ７ R ８ , or NO ２ optionally replaced by R １ , and R ２ are each independently selected from 6- to 10-membered aryl, or 9- to 18-membered benzoheterocyclyl, wherein the 6- to 10-membered aryl or 9- to 18-membered benzoheterocyclyl is selected from halogen, hydroxy, cyano, -NR ７ R ８ , NO ２ , -NR ９ C(O)R １０ , C1-C6 alkyl, (C1-C6 alkyl)-O-, -C(O)NR ９ R １０ or a 5- to 7-membered heteroaryl, wherein the C1-C6 alkyl, by itself or as part of another group or the 5- to 7-membered heteroaryl, is optionally substituted with halogen, cyano, hydroxy, C1-C3 alkyl, (C1-C3 alkyl)-O-, or -NR ７ R ８ optionally replaced by R ７ , R ８ , R ９ , and R １０ are each independently selected from H or C1-C6 alkyl; The condition is that at most two of W, X, Y, and Z are simultaneously N.

2. 2. The compound of formula I according to claim 1, wherein the compound of formula I has the structure shown in formula II: or a pharma- ceutically acceptable salt thereof. 【Chemistry 2】 (However, R １ , R ２ , R ３ , R ４ , and R ５ The definitions are the same as those in the compound of formula I.

3. R ３ , R ４ , R ５ , and R ６ are each independently hydrogen, halogen, hydroxy, NH ２ , (C1-C6 alkyl)-NR ７ -, (C1-C6 alkyl)-O-, C1-C6 alkyl, C3-C6 cycloalkyl, or 6-10 membered aryl, where the C1-C6 alkyl, by itself or as part of another group or the C3-C6 cycloalkyl, is selected from halogen, cyano, hydroxy, or -NR ７ R ８ wherein said 6-10 membered aryl is optionally substituted by halogen substituted C1-C3 alkoxy; Preferably, R ３ , R ４ , R ５ , and R ６ are each independently hydrogen, halogen, hydroxy, NH ２ , (C1-C6 alkyl)-NR ７ -, (C1-C6 alkyl)-O-, C1-C6 alkyl, C3-C6 cycloalkyl, or 6-10 membered aryl, wherein the C1-C6 alkyl is optionally substituted by halogen, either by itself or as part of another group or as C3-C6 cycloalkyl, and the 6-10 membered aryl is optionally substituted by halogen-substituted C1-C3 alkoxy; More preferably, R ３ , R ４ , R ５ , and R ６ are each independently hydrogen, halogen, hydroxy, NH ２ , (C1-C6 alkyl)-NR ７ The compound according to claim 1 or 2, characterized in that it is selected from -, (C1-C6 alkyl)-O-, C1-C6 alkyl, C3-C6 cycloalkyl, or 6-10 membered aryl, wherein the C1-C6 alkyl is optionally substituted by fluorine, either by itself or as part of another group or as C3-C6 cycloalkyl, and the 6-10 membered aryl is optionally substituted by fluorine-substituted C1-C3 alkoxy, or a pharma- ceutically acceptable salt thereof.

4. R ３ is selected from hydrogen, C1-C6 alkyl, or C3-C6 cycloalkyl, preferably R ３ is selected from hydrogen or C1-C6 alkyl, more preferably R ３ is selected from hydrogen, methyl or cyclopropyl, more preferably R ３ is selected from hydrogen or methyl, most preferably R ３ The compound according to any one of claims 1 to 3, or a pharma- ceutically acceptable salt thereof, wherein is selected from hydrogen.

5. R ４ is selected from hydrogen or C1-C6 alkyl, preferably R ４ is selected from hydrogen or methyl, more preferably R ４ The compound according to any one of claims 1 to 4, or a pharma- ceutically acceptable salt thereof, wherein is selected from hydrogen.

6. R ５ is hydrogen, halogen, hydroxy, (C1-C6 alkyl)-NR ７ -, (C1-C6 alkyl)-O-, C3-C6 cycloalkyl, or 6-10 membered aryl, wherein the C1-C6 alkyl is optionally substituted by fluorine, either by itself or as part of another group or as C3-C6 cycloalkyl, and the 6-10 membered aryl is optionally substituted by fluorine-substituted C1-C3 alkoxy, preferably R ５ is hydrogen, chlorine, hydroxy, cyclopropyl, CF ３ CH ２ O-, CHF ２ O-, CF ３ CH ２ NH-, 4-difluoromethoxyphenyl, or CH ３ CH ２ O-, more preferably R ５ is cyclopropyl, CF ３ CH ２ O-, CF ３ CH ２ NH- or CH ３ CH ２ O-, more preferably R ５ is cyclopropyl, CF ３ CH ２ O- or CF ３ CH ２ NH-, most preferably R ５ The compound according to any one of claims 1 to 5, or a pharma- ceutically acceptable salt thereof, wherein is selected from cyclopropyl.

7. R １ , and R ２ are each independently selected from phenyl and a group represented by the formula: 【Chemistry 3】 wherein the group is halogen, hydroxy, cyano, -NR ７ R ８ , NO ２ , -NR ９ C(O)R １０ , C1-C6 alkyl, (C1-C6 alkyl)-O-, -C(O)NR ９ R １０ or a 5- to 7-membered heteroaryl, wherein the C1-C6 alkyl, by itself or as part of another group or the 5- to 7-membered heteroaryl, is optionally substituted with halogen, cyano, hydroxy, C1-C3 alkyl, (C1-C3 alkyl)-O-, or -NR ７ R ８ Optionally substituted by, preferably by R １ , and R ２ are each independently selected from phenyl and a group represented by the formula: 【Chemistry 4】 wherein said group is optionally substituted by C1-C6 alkyl, or (C1-C6 alkyl)-O-, said C1-C6 alkyl being optionally substituted by halogen, either by itself or as part of another group, preferably R ７ , R ８ , R ９ , and R １０ or a pharma- ceutically acceptable salt thereof according to any one of claims 1 to 6, wherein each of

8. R １ is selected from phenyl, a group represented by the formula: 【Chemistry 5】 wherein said group is optionally substituted by C1-C6 alkyl, or (C1-C6 alkyl)-O-, said C1-C6 alkyl being optionally substituted by halogen, either by itself or as part of another group, preferably R １ is selected from the groups represented by the formula: 【Chemistry 6】 More preferably, R １ The compound according to any one of claims 1 to 7, wherein is selected from the group represented by the following formula: or a pharma- ceutically acceptable salt thereof. 【Chemistry 7】

9. R ２ is selected from phenyl or a group represented by the formula: 【Chemistry 8】 wherein said group is optionally substituted by (C1-C6 alkyl)-O-, said C1-C6 alkyl being optionally substituted by halogen, preferably R ２ is selected from the groups represented by the formula: 【Chemistry 9】 More preferably, R ２ The compound according to any one of claims 1 to 8, wherein is selected from the group represented by the following formula: or a pharma- ceutically acceptable salt thereof. 【Chemistry 10】

10. The following compound or a pharma- ceutically acceptable salt thereof: 【Chemistry 11】

11. A pharmaceutical composition comprising a therapeutically effective amount of a compound according to any one of claims 1 to 10 or a pharma- ceutically acceptable salt thereof.

12. Use of a compound according to any one of claims 1 to 10, a pharma- ceutically acceptable salt thereof or a pharmaceutical composition according to claim 11 in the preparation of a medicament for the prevention and / or treatment of a MAT2A mediated disease or condition, preferably said MAT2A mediated disease or condition being mediated by MAT2A overexpression.

13. 1. A method for preventing and / or treating a MAT2A mediated disease or condition, comprising: A method comprising administering to an individual in need thereof an effective amount of a compound according to any one of claims 1 to 10, a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition according to claim 11.

14. A compound according to any one of claims 1 to 10, a pharma- ceutically acceptable salt thereof or a pharmaceutical composition according to claim 11 for the prevention and / or treatment of a MAT2A mediated disease or condition.

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