RNA methyltransferase inhibitor, screening method of the same, effectiveness determination marker of anticancer agent, and kit for effectiveness prediction of ftsj1 inhibitor

Abstract

To provide an RNA methyltransferase inhibitor for treating cancer.SOLUTION: An RNA methyltransferase inhibitor contains a pyrazoline-based compound represented by general formula (2). [In the formula (2), n denotes an integer of 2 to 4, R4 denotes, identically or differently, a phenyl group, a phenylsulfonyl group, an alkylcarbonyl group, an aminothiocarbonyl group, a benzodioxolyl group, an alkylsulfonyl group, an adamanthylcarbonyl group, a benzopyrazyl group or the like, and each group further may have a substituent. The bond between the fourth-position carbon atom and the fifth-position carbon atom of a pyrazole skeleton is a single bond or double bond. Alternately, two adjacent carbon atoms constituting a pyrazoline ring may bind to each other to form a ring. Alternatively, a nitrogen atom and a carbon atom adjacent to the atom constituting a pyrazoline ring may bind to each other to form a ring.]SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to an RNA methyltransferase inhibitor, a screening method therefor, a marker for determining the efficacy of an anticancer drug, and a kit for predicting the efficacy of an FTSJ1 inhibitor. [Background technology]

[0002] For example, in the field of anticancer drugs, anticancer drugs with a mechanism of action related to cell cycle activity have been proposed from the viewpoint of suppressing tumor tissue hypertrophy by inhibiting the proliferation of cancer cells.

[0003] However, many cancer cells have extremely long cell cycles or are in the resting phase of the cell cycle, and conventional anticancer drugs with mechanisms of action related to cell cycle activity are unable to effectively exert their anticancer effects on these cancer cells. Summary of the Invention [Problem to be solved by the invention]

[0004] With the recent advances in nucleic acid analysis techniques, it has become clear that translational forms based on specific nucleic acid modifications play an important role in maintaining the functions of cells and viruses.

[0005] For example, in cancer cells, the translational forms specific to cancer cells are being elucidated, and it is becoming clear that these translational forms play a major role in various phenomena such as cancer cell survival, proliferation, metastasis, and maintenance of stemness. Enzymes involved in the cellular translation function, such as RNA methyltransferase, are attracting attention.

[0006] In view of the above circumstances, an object of the present invention is to provide an RNA methyltransferase inhibitor and a screening method therefor. [Means for solving the problem]

[0007] The present inventors have conducted extensive research to solve the above problems and have found that at least one compound selected from the group consisting of sulfonamide compounds represented by the following general formula (1) and pyrazoline compounds represented by the following general formula (2) has the desired RNA methyltransferase inhibitory activity. The present invention was completed based on this finding.

[0008] That is, the present invention provides the following RNA methyltransferase inhibitors, novel sulfonamide compounds, screening methods, markers for determining the effectiveness of anticancer drugs, and kits for predicting the effectiveness of FTSJ1 inhibitors.

[0009] Section 1. An RNA methyltransferase inhibitor comprising at least one compound selected from the group consisting of sulfonamide compounds represented by the following general formula (1) and pyrazoline compounds represented by the following general formula (2):

[0010] [ka]

[0011] [In formula (1), R 1 represents any one of the following groups (1-1) to (1-5). (1-1) A nitrogen-containing heterocyclic group which may have a substituent, (1-2) a cycloalkyl group which may have a substituent; (1-3) an alkyl group which may have a substituent; (1-4) pyrazolylamino group, (1-5) phenyl group; R 2 represents (2-1) a hydrogen atom or (2-2) an alkyl group. R 3 represents any one of the following groups (3-1) to (3-9). (3-1) phenyl group, (3-2) naphthyl group, (3-3) nitrogen- or sulfur-containing heterocyclic groups, (3-4) dihydrocarbostyril group, (3-5) tetrahydronaphthyl group, (3-6) indanyl group, (3-7) benzoxolyl group, (3-8) benzothiadiazolyl group, (3-9) Dihydrobenzodioxepinyl group; Each of the groups (3-1) to (3-9) may further have a substituent. 1 and R 2 may be bonded to the nitrogen atom to which they are bonded to form a ring.

[0012] [ka]

[0013] [In formula (2), n represents an integer of 2 to 4. R 4 are the same or different and represent any of the following groups (4-1) to (4-34). (4-1) phenyl group, (4-2) phenylsulfonyl group, (4-3) alkylcarbonyl group, (4-4) aminothiocarbonyl group, (4-5) benzodioxolyl group, (4-6) alkylsulfonyl group, (4-7) Adamantyl carbonyl group, (4-8) benzopyrazyl group, (4-9) phenylcarbonyl group, (4-10) naphthyl group, (4-11) Furylcarbonyl group, (4-12) thienylcarbonyl group, (4-13) quinazolyl group, (4-14) quinoxalyl group, (4-15) Hydroxyl group, (4-16) an alkenyl group, (4-17) Thiazolyl group, (4-18) cycloalkylcarbonyl group, (4-19) Aminocarbonyl group, (4-20) Furyl group, (4-21) thienyl group, (4-22) Pyridyl group, (4-23) a cycloalkenyl group, (4-24) alkyl group, (4-25) Pyrazolyl group, (4-26) quinolyl group, (4-27) Alkenylcarbonyl group, (4-28) Benzopyranyl group, (4-29) Benzopyrimidyl group, (4-30) pyrrolidinoalkylcarbonyl group, (4-31) quinolylcarbonyl group, (4-32) Alkoxycarbonyl group, (4-33) Morpholino group, (4-34) pyrrolidinocarbonylalkoxy group, (4-35) benzodioxy-6-yl group; Each of these groups (4-1) to (4-35) may further have a substituent. The bond between the 4-position carbon atom and the 5-position carbon atom of the pyrazole skeleton represents a single bond or a double bond. Alternatively, two adjacent carbon atoms constituting the pyrazoline ring may be bonded to each other to form a ring. Alternatively, the nitrogen atom constituting the pyrazoline ring and the carbon atom adjacent thereto may be bonded to each other to form a ring.

[0014] Section 2. 2. The RNA methyltransferase inhibitor according to claim 1, wherein the substituent on the nitrogen-containing heterocyclic group represented by (1-1) is at least one selected from the group consisting of an alkyl group, a hydroxyl group, a cyclopropyl group, a phenylthiopropylcarbonyl group, a phenylsulfonyl group, an alkylsulfonyl group, a thienylsulfonyl group, an alkylcarbonyl group, an alkoxycarbonyl group, a phenylsulfonylamino group, an aminocarbonylalkyl group, a pyrazolylcarbonyl group, a cyclopropylcarbonyl group, a piperidylsulfonyl group, and a morpholinosulfonyl group.

[0015] Section 3. Item 2. The RNA methyltransferase inhibitor according to Item 1, wherein the nitrogen-containing heterocyclic group represented by (1-1) has 1 to 5 substituents.

[0016] Section 4. Item 4. The RNA methyltransferase inhibitor according to Item 2 or 3, wherein the alkyl group and alkoxy group moieties constituting the alkyl group, alkylsulfonyl group, alkylcarbonyl group, alkoxycarbonyl group, and aminocarbonylalkyl group on the nitrogen-containing heterocyclic group represented by (1-1) have 1 to 4 carbon atoms.

[0017] Section 5. Item 5. The RNA methyltransferase inhibitor according to any one of Items 2 to 4, wherein the phenylsulfonyl group on the nitrogen-containing heterocyclic group represented by (1-1) further has at least one substituent selected from the group consisting of a halogen atom, an alkyl group, a fluoroalkyl group, an alkoxy group, and a nitro group.

[0018] Section 6. Item 2. The RNA methyltransferase inhibitor according to Item 1, wherein the cycloalkyl group represented by (1-2) has 3 to 6 carbon atoms.

[0019] Section 7. Item 2. The RNA methyltransferase inhibitor according to Item 1, wherein the cycloalkyl group represented by (1-2) has 5 or 6 carbon atoms.

[0020] Section 8. Item 2. The RNA methyltransferase inhibitor according to Item 1, wherein the alkyl group represented by (1-3) is a linear alkyl group having 1 to 6 carbon atoms which may have a substituent.

[0021] Section 9. Item 9. The RNA methyltransferase inhibitor according to any one of Items 1 to 8, wherein the substituent on the cycloalkyl group represented by (1-2) and the substituent on the alkyl group represented by (1-3) are at least one selected from the group consisting of a phenyl group, a biphenyl group, a cycloalkyl group, a cycloalkenyl group, a nitrogen-containing heterocyclic group, and a hydroxyl group.

[0022] Section 10. Item 10. The RNA methyltransferase inhibitor according to any one of Items 1 to 9, wherein the phenyl group, biphenyl group, cycloalkyl group, cycloalkenyl group, nitrogen-containing heterocyclic group, or hydroxyl group present on the alkyl group represented by (1-3) further has an alkyl group having 1 to 5 carbon atoms as a substituent.

[0023] Section 11. The RNA methyltransferase inhibitor according to claim 1, wherein the substituent on each of the groups (3-1) to (3-9) is at least one selected from the group consisting of an alkyl group, an alkoxy group, a halogen atom, a carboxyl group, an amino group, a nitro group, a phenyl group, and a cycloalkyl group.

[0024] Section 12. Item 2. The RNA methyltransferase inhibitor according to Item 1, wherein the number of substituents on each of the groups (3-1) to (3-9) is 1 to 5.

[0025] Section 13. Item 2. The RNA methyltransferase inhibitor according to Item 1, wherein the alkyl group and alkoxy group on the phenyl group represented by (3-1) have 1 to 5 carbon atoms, and the cycloalkyl group has 3 to 7 carbon atoms.

[0026] Section 14. Item 2. The RNA methyltransferase inhibitor according to Item 1, wherein the number of substituents on each of the groups (4-1) to (4-35) is 1 to 6.

[0027] Section 15. Item 2. The RNA methyltransferase inhibitor according to Item 1, wherein the substituent on each of the groups (4-1) to (4-35) is at least one selected from the group consisting of a linear or branched alkyl group, a cycloalkyl group, an alkoxy group, an alkylamino group, a phenyl group, a phenylalkyl group, a phenylalkenyl group, a halogen atom, a nitro group, a carboxy group, a furyl group, a dihydroxyphenyl group, a biphenylyl group, an alkylcarbonyl group, an oxo-substituted quinolyl group, a benzofuranyl group, a thienyl group, a trialkylamino group, an oxo group, and a pyridyl group.

[0028] Section 16. Item 2. The RNA transferase inhibitor according to Item 1, wherein the substituent on the phenyl group represented by (4-1) is at least one selected from the group consisting of halogen, alkyl group, haloalkyl group, alkoxy group, hydroxyl group, alkylsulfonylamino group, nitro group, amino group, carboxyl group, and phenyl group.

[0029] Section 17. The substituent on the alkylcarbonyl group represented by (4-3) above is a phenylalkylamino group, a triazolylthio group, a phenoxy group, an oxadiazolylthio group, an ester group, a piperazinyl group, a carboxyl group, a pyrimidinylthio group, a quinazolyloxy group, a morpholinocarbonyl group, a morpholino group, a benzotriazolyl group, a pyrazolylcarbonyl group, a pyrimidyl group, a pyrrolidino group, a piperidino group, a tetrahydroimidazolyl group, a halogen atom, a naphthyloxy group, an alkoxy group, an imidazolyl group, a tetrazolylthio group, an alkylamino group, a pyridyl group, a tetrazolyl group, a benzodioxonyloxy group, an aminocarbonyl group, a piperazinyloxy group, a Item 2. The RNA methyltransferase inhibitor according to Item 1, wherein the RNA methyltransferase inhibitor is at least one selected from the group consisting of an alkyl group, a phenylalkylthio group, an alkylcarbonyloxy group, a benzotriazolylthio group, a pyridazinyl group, a pyrrolylcarbonyloxy group, a piperidino group, a dihydrothiazolylthio group, a benzopyrazyl group, a thienopyridinoxy group, a thienopyrimidinylthio group, a cyclopentathienopyrimidinyl group, a thiadiazolylthio group, an azepinylthio group, a dioxoloquinolinyl group, a diazaspirononanyl group, an imidazolidinyl group, a triazolylthio group, a dihydropyridazinyl group, and a 1,3-diazaspironoundecanyl group.

[0030] Section 18. Item 2. The RNA methyltransferase inhibitor according to Item 1, wherein each group on the alkylcarbonyl group represented by (4-3) may further have 1 to 6 substituents.

[0031] Section 19. Item 2. The RNA methyltransferase inhibitor according to Item 1, wherein each group on the alkylcarbonyl group represented by (4-3) further has at least one substituent selected from the group consisting of a linear, branched, or cyclic alkyl group, an alkoxy group, an alkoxyphenyl group, an amino group, a carbamoyl group, a carbamoylalkyl group, a thienyl group, a furyl group, a tetrazolyl group, an alkylcarbonyl group, a halogen atom, a phenyl group, a furanyl group, an alkylpyrrolidinyl group, a thiophenyl group, a furylcarbonyl group, an oxo group, a trifluoroalkyl group, a hydroxyl group, a thienylalkyl group, an alkylaminosulfonyl group, a hydroxyalkyl group, a furanylcarbonyl group, a benzylthio group, a nonanyl group, a bicyclononanyl group, an alkylthiadiazolyl group, and an alkylisoxazolyl group.

[0032] Section 20. Item 2. The RNA methyltransferase inhibitor according to Item 1, wherein in the general formula (2), the ring formed by bonding two adjacent carbon atoms constituting the pyrazoline ring to each other is a cyclohexane ring.

[0033] Section 21. Item 21. The RNA methyltransferase inhibitor according to Item 20, wherein the cyclohexane ring has a vinyl group which may have a substituent.

[0034] Section 22. Item 22. The RNA methyltransferase inhibitor according to Item 21, wherein the substituent on the vinyl group is at least one selected from the group consisting of a phenyl group, a benzoxonyl group, a furyl group, a thienyl group, and a cyclopentane ring.

[0035] Section 23. Item 2. The RNA methyltransferase inhibitor according to Item 1, wherein in the general formula (2), the nitrogen atom constituting the pyrazoline ring and the carbon atom adjacent thereto are bonded to each other to form a cyclohexane ring which may have a substituent.

[0036] Section 24. Item 2. The RNA methyltransferase inhibitor according to Item 1, wherein in general formula (2), the nitrogen atom constituting the pyrazoline ring and the carbon atom adjacent thereto are bonded to each other to form a cyclohexane ring which may have a substituent, and the substituent on the cyclohexane ring is at least one selected from the group consisting of a halogen atom, an alkyl group, an alkoxy group, a (bi)phenyl group which may have a substituent, an alkylphenyl group, an alkoxyphenyl group, a pyridyl group, an alkoxyphenyl group, a nitrophenyl group, a (di)fluorophenyl group, a (di)chlorophenyl group, and a spiro ring.

[0037] Section 25. In the general formula (2), a nitrogen-containing heterocyclic group on a nitrogen atom constituting the pyrazoline ring; Item 2. The RNA methyltransferase inhibitor according to Item 1, wherein the hydroxyphenyl group on the carbon atom adjacent to the nitrogen atom constituting the pyrazoline ring is bonded to each other to form a ring.

[0038] Section 26. Item 26. The RNA transferase inhibitor according to any one of Items 1 to 25, which is used in the treatment of cancer.

[0039] Section 27. A sulfonamide compound represented by the following general formula (1a):

[0040] [ka]

[0041] [In formula (1a), R 1a represents a piperidyl group which may have a substituent, a pyridyl group which may have a substituent, a pyrazolyl group which may have a substituent, a cyclohexyl group, a linear alkyl group having 1 to 5 carbon atoms which may have a substituent, a pyrazolylamino group which may have a substituent, or a phenylamino group which may have a substituent. R 2a represents a hydrogen atom or a methyl group. 3arepresents a phenyl group which may have a substituent.

[0042] Section 28. R 1a Item 28. The sulfonamide compound according to Item 27, wherein the number of substituents on the piperidyl group, the pyridyl group, the pyridyl group, the pyrazolyl group, the cyclohexyl group, the linear alkyl group having 1 to 5 carbon atoms, the pyrazolylamino group, or the phenylamino group is 1 to 5, and is represented by the following formula:

[0043] Section 29. R 1a Item 28. The sulfonamide compound according to Item 27, wherein the substituent on the piperidyl group represented by the formula: is at least one selected from the group consisting of a methyl group and a hydroxyl group.

[0044] Section 30. R 1a Item 28. The sulfonamide compound according to Item 27, wherein the substituent on the linear alkyl group having 1 to 5 carbon atoms, represented by the formula: is at least one selected from the group consisting of a cycloalkyl group, a cycloalkenyl group, a nitrogen-containing heterocyclic group, and a hydroxyl group.

[0045] Section 31. R 1a Item 28. The sulfonamide compound according to Item 27, wherein the substituent on the linear alkyl group having 1 to 5 carbon atoms, represented by the formula: is at least one selected from the group consisting of a cyclohexyl group, a cyclohexenyl group, a piperidyl group, and a hydroxyl group.

[0046] Section 32. R 3a Item 28. The sulfonamide compound according to item 27, wherein the number of substituents on the phenyl group is 1 to 3.

[0047] Section 33. R 3aItem 28. The sulfonamide compound according to Item 27, wherein the substituent on the phenyl group represented by the formula: is at least one selected from the group consisting of an alkyl group having 1 to 6 carbon atoms, an alkoxy group having 1 to 3 carbon atoms, a phenyl group, a halogen atom, and a carboxyl group.

[0048] Section 34. R 3a Item 28. The sulfonamide compound according to Item 27, wherein the substituent on the phenyl group represented by the formula: is at least one selected from a methyl group, an isopropyl group, a tert-butyl group, a tert-pentyl group, a methoxy group, a phenyl group, a cyclopropyl group, a chlorine atom, and a carboxyl group.

[0049] Section 35. A method for screening RNA transferase inhibitors, comprising a step of measuring the inhibitory effect of a test substance on RNA methylation in cells or viruses.

[0050] Section 36. Item 36. The method according to Item 35, wherein the RNA methylation inhibitory effect is based on FTSJ inhibition.

[0051] Section 37. Item 36. The method of item 35, wherein the FTSJ is FTSJ1.

[0052] Section 38. Item 38. The method according to any one of Items 35 to 37, wherein the RNA methylation inhibitory effect is measured by a reporter assay using a sequence in which a translation regulatory region is added to a reporter region, The method according to claim 1, wherein the translation regulatory region comprises a sequence formed by the binding of at least one selected from the group consisting of glutamine, phenylalanine, tryptophan, methionine, and leucine.

[0053] Section 39. Item 39. The method according to Item 38, wherein the translation regulatory region comprises a sequence in which 5 to 50 consecutively linked amino acids of at least one selected from the group consisting of glutamine, phenylalanine, tryptophan, methionine, and leucine are linked.

[0054] Section 40. Item 39. The method according to Item 38, wherein the translation regulatory region consists of polyglutamine, polyphenylalanine, polytryptophan, polymethionine, or polyleucine, each of which has 5 to 50 consecutively linked glutamines, phenylalanines, tryptophans, methionines, or leucines.

[0055] Section 41. Item 39. The method according to Item 38, wherein the translation regulatory region is any one of SEQ ID NOs: 1 to 12.

[0056] Section 42. 42. The method according to any one of items 35 to 41, further comprising carrying out a reporter assay using a sequence comprising the transcription factor binding region shown in SEQ ID NO: 13 and a reporter region.

[0057] Section 43. Adding a methyl group donor to the test substance to obtain a reaction product; and a step of measuring the FTSJ1 activity of the test substance using the reaction product, in this order;

[0058] Section 44. Item 44. The method of Item 43, wherein the methyl group donor is S-adenosylmethionine (SAM).

[0059] Section 45. Item 45. The method according to Item 44, wherein the FTSJ1 activity measurement is carried out by luciferase assay.

[0060] Section 46. A method for predicting the effectiveness of an FTSJ1 inhibitor against cancer, or a method for predicting the prognosis of cancer by use of an FTSJ1 inhibitor, comprising step A of measuring the expression level of FTSJ1 in a sample.

[0061] Section 47. Item 47. The method according to Item 46, wherein step A is carried out by an immunological technique or a genetic technique.

[0062] Section 48. 48. The method of claim 46 or 47, wherein the sample is taken from a patient.

[0063] Section 49. Item 49. The method according to any one of Items 46 to 48, further comprising Step B of determining the efficacy of an FTSJ1 inhibitor against cancer or determining the prognosis of the patient's cancer pathology based on the FTSJ1 expression level obtained in Step A.

[0064] Section 50. Item 50. The method according to any one of Items 46 to 49, wherein the cancer is at least one selected from the group consisting of glioblastoma (malignant brain tumor), pancreatic cancer, acute myeloid leukemia, lung cancer, liver cancer, kidney cancer, stomach cancer, and breast cancer.

[0065] Section 51. A marker for determining the effectiveness of anticancer drugs, consisting of a gene marker associated with sensitivity to FTSJ1 inhibitors or a gene marker associated with resistance to FTSJ1 inhibitors.

[0066] Section 52. Item 52. The marker according to Item 51, wherein the FTSJ1 inhibitor sensitivity-associated gene marker or the FTSJ1 inhibitor resistance-associated gene marker is an FTSJ1 modified nucleic acid RNA.

[0067] Section 53. The FTSJ1 inhibitor resistance-associated gene markers include AHNAK nucleoprotein 2 (AHNAK2, SEQ ID NO: 14), extended synaptotagmin 1 (ESYT1, SEQ ID NO: 15), SLIT-ROBO Rho GTPase activating protein 1 (SRGAP1, SEQ ID NO: 16), ras homolog family member F, filopodia associated (RHOF, SEQ ID NO: 17), microRNA 4746 (MIR4746, SEQ ID NO: 18), UBX domain protein 6 (UBXN6, SEQ ID NO: 19), cytochrome c oxidase assembly factor COX16 (COX16, SEQ ID NO: 20), ferritin heavy chain 1 (FTH1, SEQ ID NO: 21), lysophosphatidic acid receptor 1 (LPAR1, SEQ ID NO: 22), ankyrin repeat domain 29 (ANKRD29, SEQ ID NO: 23), and twist family bHLH transcription factor 2 (TWIST2, SEQ ID NO: 24), JNK1 / MAPK8 associated membrane protein (JKAMP, SEQ ID NO: 25), protein kinase AMP-activated catalytic subunit alpha 2 (PRKAA2, SEQ ID NO: 26), cleavage stimulation factor subunit 2 tau variant (CSTF2T, SEQ ID NO: 27), thrombospondin type 1 domain containing 4 (THSD4, SEQ ID NO: 28), membrane associated guanylate kinase,Item 51. The marker according to Item 50, which is at least one selected from the group consisting of WW and PDZ domain containing 1 (MAGI1, SEQ ID NO: 29), ubiquitin conjugating enzyme E2 L3 (UBE2L3, SEQ ID NO: 30), glycosylphosphatidylinositol specific phospholipase D1 (GPLD1, SEQ ID NO: 31), FRY-like transcription coactivator (FRYL, SEQ ID NO: 32), and myosin IXA (MYO9A, SEQ ID NO: 33).

[0068] Section 54. The gene markers associated with sensitivity to FTSJ1 inhibitors include RNA binding motif protein 15 (RBM15, SEQ ID NO: 34), nuclear autoantigenic sperm protein (NASP, SEQ ID NO: 35), pre-mRNA processing factor 38A (PRPF38A, SEQ ID NO: 36), chromosome 1 open reading frame 50 (C1orf50, SEQ ID NO: 37), peroxisomal biogenesis factor 16 (PEX16, SEQ ID NO: 38), zinc finger protein 213 (ZNF213, SEQ ID NO: 39), fem-1 homolog B (FEM1B, SEQ ID NO: 40), regulatory factor X associated protein (RFXAP, SEQ ID NO: 41), Sin3A associated protein 18 (SAP18, SEQ ID NO: 42), alanyl-tRNA synthetase 2, mitochondrial (AARS2, SEQ ID NO: 43), regulator of chromosome condensation 2 (RCC2, SEQ ID NO: 44), and tyrosyl-tRNA synthetase 1 (YARS1, SEQ ID NO: 45), RNA binding motif protein 10 (RBM10, SEQ ID NO: 46), ribosomal protein L5 (RPL5, SEQ ID NO: 47), zinc finger HIT-type containing 2 (ZNHIT2, SEQ ID NO: 48), oxidative stress-induced growth inhibitor family member 2 (OSGIN2, SEQ ID NO: 49), egl-9 family hypoxia inducible factor 3 (EGLN3, SEQ ID NO: 50), tRNA phosphotransferase 1 (TRPTI, SEQ ID NO: 51), CRACD-like (CRACDL, SEQ ID NO: 52), capping actin protein,Item 50. The marker according to Item 50, which is at least one selected from the group consisting of gelsolin-like (CAPG, SEQ ID NO: 53), RAB11 family interacting protein 3 (RAB11FIP3, SEQ ID NO: 54), calcium homeostasis modulator family member 5 (CALHM5, SEQ ID NO: 55), BICD cargo adaptor 1 (BICD1, SEQ ID NO: 56), and FtsJ RNA 2'-O-Methyltransferase 1 (FTSJ1, SEQ ID NO: 57).

[0069] Section 55. Item 55. A kit for predicting the efficacy of an FTSJ1 inhibitor, comprising the marker according to any one of Items 51 to 54. [Effects of the Invention]

[0070] The present invention provides an RNA methyltransferase inhibitor, a screening method therefor, a marker for determining the efficacy of an anticancer drug, and a kit for predicting the efficacy of an FTSJ1 inhibitor. [Brief explanation of the drawings]

[0071] [Figure 1] 1 shows the results of animal experiments on the RNA methyltransferase inhibitor of the present invention. [Figure 2] 1 shows the results of animal experiments on the RNA methyltransferase inhibitor of the present invention. [Figure 3] 1 shows the results of detecting FTSJ1 inhibitory activity using the screening method of the present invention. [Figure 4] Evaluation results of genetic markers related to sensitivity to FTSJ1 inhibitors. DETAILED DESCRIPTION OF THE INVENTION

[0072] 1. RNA methyltransferase inhibitors The RNA methyltransferase inhibitor of the present invention includes a compound represented by the following general formula (1) and / or a pyrazoline compound represented by the following general formula (2).

[0073] [ka]

[0074] [In formula (1), R 1 represents any one of the following groups (1-1) to (1-5). (1-1) A nitrogen-containing heterocyclic group which may have a substituent, (1-2) a cycloalkyl group which may have a substituent; (1-3) an alkyl group which may have a substituent; (1-4) pyrazolylamino group, (1-5) phenyl group; R 2 represents (2-1) a hydrogen atom or (2-2) an alkyl group. R 3 represents any one of the following groups (3-1) to (3-9). (3-1) phenyl group, (3-2) naphthyl group, (3-3) nitrogen- or sulfur-containing heterocyclic groups, (3-4) dihydrocarbostyril group, (3-5) tetrahydronaphthyl group, (3-6) indanyl group, (3-7) benzoxolyl group, (3-8) benzothiadiazolyl group, (3-9) Dihydrobenzodioxepinyl group; Each of the groups represented by (3-1) to (3-9) may further have a substituent. Or, R 1 and R 2 may be bonded to the nitrogen atom to which they are bonded to form a ring.

[0075] [ka]

[0076] [In formula (2), n represents an integer of 2 to 4. R4 are the same or different and represent any of the following groups (4-1) to (4-35). (4-1) phenyl group, (4-2) phenylsulfonyl group, (4-3) alkylcarbonyl group, (4-4) aminothiocarbonyl group, (4-5) benzodioxolyl group, (4-6) alkylsulfonyl group, (4-7) Adamantyl carbonyl group, (4-8) benzopyrazyl group, (4-9) phenylcarbonyl group, (4-10) naphthyl group, (4-11) Furylcarbonyl group, (4-12) thienylcarbonyl group, (4-13) quinazolyl group, (4-14) quinoxalyl group, (4-15) Hydroxyl group, (4-16) an alkenyl group, (4-17) Thiazolyl group, (4-18) cycloalkylcarbonyl group, (4-19) Aminocarbonyl group, (4-20) Furyl group, (4-21) thienyl group, (4-22) Pyridyl group, (4-23) a cycloalkenyl group, (4-24) alkyl group, (4-25) Pyrazolyl group, (4-26) quinolyl group, (4-27) Alkenylcarbonyl group, (4-28) Benzopyranyl group, (4-29) Benzopyrimidyl group, (4-30) pyrrolidinoalkylcarbonyl group, (4-31) quinolylcarbonyl group, (4-32) Alkoxycarbonyl group, (4-33) Morpholino group, (4-34) pyrrolidinocarbonylalkoxy group, (4-35) benzodioxy-6-yl group; Each of the groups (4-1) to (4-35) may further have a substituent. The bond between the nitrogen atom at position 2 and the carbon atom at position 3 of the pyrazole skeleton represents a single bond or a double bond. Alternatively, two adjacent carbon atoms constituting the pyrazoline ring may be bonded to each other to form a ring. Alternatively, the nitrogen atom constituting the pyrazoline ring and the carbon atom adjacent thereto may be bonded to each other to form a ring.

[0077] The present inventors have found that the compounds represented by the above general formulas (1) and (2) inhibit the methylation modification reaction of RNA by competitively binding with S-adenosylmethionine (hereinafter also referred to simply as SAM) to the region (hereinafter also referred to simply as SAM binding region) of FTSJ, a tRNA methylation modification enzyme.

[0078] Furthermore, the present inventors have found that the compounds represented by general formulas (1) and (2) have antitumor activity based on the above-mentioned inhibitory activity against RNA methylation and are useful as therapeutic agents for cancer. In this specification, the term "therapeutic agents for cancer" is defined to include not only those generally called anticancer agents but also agents that suppress cancer metastasis.

[0079] The compounds represented by general formulas (1) and (2) are described in detail below.

[0080] In this specification, examples of the alkyl group include alkyl groups having 1 to 6 carbon atoms, specifically, methyl groups, ethyl groups, n-propyl groups, isopropyl groups, n-butyl groups, isobutyl groups, sec-butyl groups, tert-butyl groups, n-pentyl groups, and n-hexyl groups.

[0081] In this specification, examples of the alkoxy group include alkoxy groups having 1 to 6 carbon atoms, specifically, methoxy, ethoxy, n-propyloxy, isopropyloxy, n-butyloxy, isobutyloxy, sec-butyloxy, tert-butyloxy, n-pentyloxy, and n-hexyloxy groups.

[0082] In this specification, the cycloalkyl group includes a cycloalkyl group having 3 to 8 carbon atoms, specifically, a cyclopropyl group, a cyclobutyl group, a cyclopentyl group, a cyclohexyl group, a cycloheptyl group, etc.

[0083] In this specification, the cycloalkenyl group refers to a cycloalkenyl group having 3 to 8 carbon atoms, specifically, a cyclopropenyl group, a cyclobutenyl group, a cyclopentenyl group, a cyclohexenyl group, a cycloheptenyl group, etc.

[0084] In general formula (1), R 1 Examples of the nitrogen-containing heterocyclic group represented by the formula include a piperidyl group, a pyridyl group, a pyrazolyl group, etc. A preferred nitrogen-containing heterocyclic group is a piperidyl group.

[0085] Substituents on the nitrogen-containing heterocyclic group include alkyl groups, hydroxyl groups, cyclopropyl groups, phenylthiopropylcarbonyl groups, phenylsulfonyl groups, alkylsulfonyl groups, thienylsulfonyl groups, alkylcarbonyl groups, alkoxycarbonyl groups, phenylsulfonylamino groups, aminocarbonylalkyl groups, pyrazolylcarbonyl groups, cyclopropylcarbonyl groups, piperidylsulfonyl groups, and morpholinosulfonyl groups. The preferred substituents on the nitrogen-containing heterocyclic group are alkyl groups, and more preferably isopropyl groups. The number of substituents is 1 to 5, preferably 1 to 4.

[0086] The alkyl group moiety and alkoxy group moiety constituting the alkyl group, alkylsulfonyl group, alkylcarbonyl group, alkoxycarbonyl group and aminocarbonylalkyl group on the nitrogen-containing heterocyclic group represented by (1-1) above each have 1 to 4 carbon atoms.

[0087] The phenylsulfonyl group on the nitrogen-containing heterocyclic group represented by (1-1) further has at least one substituent selected from the group consisting of a halogen atom, an alkyl group, a fluoroalkyl group, an alkoxy group and a nitro group.

[0088] In general formula (1), R 1 Examples of the cycloalkyl group represented by the formula (I) include a cycloalkyl group having 3 to 8 carbon atoms, specifically a cyclopropyl group, a cyclobutyl group, a cyclopentyl group, a cyclohexyl group, a cycloheptyl group, etc. The cycloalkyl group preferably has 3 to 6 carbon atoms. More preferred cycloalkyl groups are a cyclopentyl group and a cyclohexyl group.

[0089] In general formula (1), R 1 Examples of the alkyl group represented by the formula (I) include alkyl groups having 1 to 6 carbon atoms, specifically methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, n-hexyl, etc. Preferred alkyl groups are methyl, ethyl, isopropyl, etc.

[0090] The substituent on the cycloalkyl group represented by (1-2) and the substituent on the alkyl group represented by (1-3) are at least one selected from the group consisting of a phenyl group, a biphenyl group, a cycloalkyl group, a cycloalkenyl group, a nitrogen-containing heterocyclic group (e.g., a piperidyl group, a pyridyl group, a pyrazolyl group, etc.), and a hydroxyl group.

[0091] The phenyl group, biphenyl group, cycloalkyl group, cycloalkenyl group, nitrogen-containing heterocyclic group, or hydroxyl group present on the alkyl group represented by (1-3) may further have an alkyl group having 1 to 5 carbon atoms as a substituent.

[0092] The substituent on each of the groups (3-1) to (3-9) is at least one selected from the group consisting of an alkyl group, an alkoxy group, a halogen atom, a carboxyl group, an amino group, a nitro group, a phenyl group, and a cycloalkyl group. The number of substituents on each of the groups (3-1) to (3-9) is 1 to 5, preferably 1 to 3.

[0093] The alkyl group and alkoxy group on the phenyl group represented by (3-1) have 1 to 5 carbon atoms, and the cycloalkyl group has 3 to 7 carbon atoms. The alkyl group and alkoxy group preferably have 1 to 3 carbon atoms.

[0094] Examples of the nitrogen- or sulfur-containing heterocyclic group represented by (3-3) include a pyrrolyl group, a piperidyl group, a quinolyl group, and a thienyl group.

[0095] R 3 As the phenyl group, (3-1) a phenyl group or (3-2) a naphthyl group is preferred. The phenyl group is more preferably substituted with 1 to 3 alkyl groups having 1 to 5 carbon atoms, and particularly preferably substituted with 3 isopropyl groups.

[0096] In general formula (2), R 4 The number of substituents on each of the groups (4-1) to (4-35) defined above is 1 to 6, preferably 1 to 3.

[0097] The substituent on each of the groups (4-1) to (4-35) is at least one selected from the group consisting of a linear or branched alkyl group, a cycloalkyl group, an alkoxy group, an alkylamino group, a phenyl group, a phenylalkyl group, a phenylalkenyl group, a halogen atom, a nitro group, a carboxy group, a furyl group, a dihydroxyphenyl group, a biphenylyl group, an alkylcarbonyl group, an oxo-substituted quinolyl group, a benzofuranyl group, a thienyl group, a trialkylamino group, an oxo group, and a pyridyl group.

[0098] The substituent on the phenyl group represented by (4-1) is preferably at least one selected from the group consisting of halogen, alkyl group, haloalkyl group, alkoxy group, hydroxyl group, alkylsulfonylamino group, nitro group, amino group, carboxyl group, and phenyl group.

[0099] The substituent on the alkylcarbonyl group represented by (4-3) above is a phenylalkylamino group, a triazolylthio group, a phenoxy group, an oxadiazolylthio group, an ester group, a piperazinyl group, a carboxyl group, a pyrimidinylthio group, a quinazolyloxy group, a morpholinocarbonyl group, a morpholino group, a benzotriazolyl group, a pyrazolylcarbonyl group, a pyrimidyl group, a pyrrolidino group, a piperidino group, a tetrahydroimidazolyl group, a halogen atom, a naphthyloxy group, an alkoxy group, an imidazolyl group, a tetrazolylthio group, an alkylamino group, a pyridyl group, a tetrazolyl group, a benzodioxonyloxy group, an amino group, a benzotriazolyl group, a pyrimidinyl group, a quinazolyloxy group, a benzotriazolyl group, a pyrimidyl group, a pyrrolidino group, a piperidino group, a tetrahydroimidazolyl group, a halogen atom, a naphthyloxy group, an alkoxy group, an imidazolyl group, a tetrazolylthio group, an alkylamino group, a pyridyl group, a tetrazolyl group, a benzodioxonyloxy group, an amino group, a benzotriazolyl group, a benzodioxonyloxy group, an amine ... and at least one selected from the group consisting of a thienocarbonyl group, a piperazinyl group, a phenylalkylthio group, an alkylcarbonyloxy group, a benzotriazolylthio group, a pyridazinyl group, a pyrrolylcarbonyloxy group, a piperidino group, a dihydrothiazolylthio group, a benzopyrazyl group, a thienopyridinoxy group, a thienopyrimidinylthio group, a cyclopentathienopyrimidinyl group, a thiadiazolylthio group, an azepinylthio group, a dioxoloquinolinyl group, a diazaspirononanyl group, an imidazolidinyl group, a triazolylthio group, a dihydropyridazinyl group, and a 1,3-diazaspironoundecanyl group.

[0100] Each group on the alkylcarbonyl group represented by (4-3) may further have 1 to 6, preferably 1 to 3, substituents.

[0101] Each group on the alkylcarbonyl group represented by (4-3) further has at least one substituent selected from the group consisting of a linear, branched, or cyclic alkyl group, an alkoxy group, an alkoxyphenyl group, an amino group, a carbamoyl group, a carbamoylalkyl group, a thienyl group, a furyl group, a tetrazolyl group, an alkylcarbonyl group, a halogen atom, a phenyl group, a furanyl group, an alkylpyrrolidinyl group, a thiophenyl group, a furylcarbonyl group, an oxo group, a trifluoroalkyl group, a hydroxyl group, a thienylalkyl group, an alkylaminosulfonyl group, a hydroxyalkyl group, a furanylcarbonyl group, a benzylthio group, a nonanyl group, a bicyclononanyl group, an alkylthiadiazolyl group, and an alkylisoxazolyl group.

[0102] In the general formula (2), the ring formed by bonding two adjacent carbon atoms constituting the pyrazoline ring is, for example, a cyclohexane ring, which preferably has a vinyl group which may have a substituent.

[0103] The substituent on the vinyl group is at least one selected from the group consisting of a phenyl group, a benzoxonyl group, a furyl group, a thienyl group, and a cyclopentane ring.

[0104] In the general formula (2), the nitrogen atom constituting the pyrazoline ring and the carbon atom adjacent thereto are preferably bonded to each other to form a cyclohexane ring which may have a substituent. The substituent on the cyclohexane ring is preferably at least one selected from the group consisting of a halogen atom, an alkyl group, an alkoxy group, a (bi)phenyl group which may have a substituent, an alkylphenyl group, an alkoxyphenyl group, a pyridyl group, an alkoxyphenyl group, a nitrophenyl group, a (di)fluorophenyl group, a (di)chlorophenyl group, and a spiro ring.

[0105] In addition, in the general formula (2), a nitrogen-containing heterocyclic group on a nitrogen atom constituting the pyrazoline ring; On the pyrazoline ring, the hydroxyphenyl group on the carbon atom adjacent to the nitrogen atom constituting the pyrazoline ring may be bonded to each other to form a ring.

[0106] Among the groups represented by (4-1) to (4-35), (4-1) a phenyl group, (4-2) a phenylsulfonyl group, (4-3) an alkylcarbonyl group, (4-4) an aminothiocarbonyl group, (4-6) an alkylsulfonyl group, (4-11) a furylcarbonyl group, (4-12) a thienylcarbonyl group, (4-20) a furyl group, (4-21) a thienyl group, (4-22) a pyridyl group, (4-25) a pyrazolyl group, or (4-35) a benzodioxy-6-yl group is preferred.

[0107] Among the groups represented by (4-1) to (4-35), (4-1) a phenyl group, (4-2) a phenylsulfonyl group, (4-3) an alkylcarbonyl group, (4-4) an aminothiocarbonyl group, (4-6) an alkylsulfonyl group, (4-11) a furylcarbonyl group, (4-12) a thienylcarbonyl group, (4-20) a furyl group, (4-21) a thienyl group, (4-22) a pyridyl group, (4-25) a pyrazolyl group, or (4-35) a benzodioxy-6-yl group is preferred.

[0108] Among the groups (4-1) to (4-35), (4-1) a phenyl group, (4-3) an alkylcarbonyl group, (4-6) an alkylsulfonyl group, or (4-35) a benzodioxy-6-yl group is more preferred.

[0109] Among the groups (4-1) to (4-35), (4-1) a phenyl group and (4-35) a benzodioxy-6-yl group are particularly preferred.

[0110] In general formula (2), the substituent of the phenyl group represented by (4-1) may be a halogen atom such as a bromine atom, an alkoxy group such as a methoxy group, or a hydroxyl group, and is preferably a hydroxyl group. The number of substituents on the phenyl group is 1 to 5, preferably 1 to 3, and more preferably 1.

[0111] When the RNA methyltransferase inhibitor of the present invention is used as a cancer therapeutic agent, it may further contain a pharmaceutically acceptable carrier in addition to the above-mentioned compound. Pharmaceutically acceptable carriers include diluents and excipients commonly used in pharmaceutical formulations, such as fillers, extenders, binders, wetting agents, disintegrants, surfactants, and lubricants. The RNA methyltransferase inhibitor of the present invention may be formulated in any common pharmaceutical formulation, such as tablets, flash-melt tablets, pills, powders, liquids, suspensions, emulsions, granules, capsules, suppositories, injections (liquids, suspensions, etc.), troches, nasal sprays, and transdermal patches. The type of cancer to which the RNA methyltransferase inhibitor can be applied is not particularly limited, and the agent can be used against a variety of cancers.

[0112] The method of administration of the RNA methyltransferase inhibitor of the present invention is not particularly limited, and is administered in a manner appropriate to the various formulations, the patient's age, sex, and other conditions (such as the severity of the disease). For example, tablets, pills, liquids, suspensions, emulsions, granules, and capsules are administered orally. Injections are administered intravenously, either alone or mixed with common fluids such as glucose and amino acids, or alone, intramuscularly, intradermally, subcutaneously, or intraperitoneally. Suppositories are administered rectally.

[0113] 2. Sulfonamide compounds The present invention also includes an invention relating to a novel sulfonamide compound, which is represented by the following general formula (1a):

[0114] [ka]

[0115] [In formula (1a), R1a represents a piperidyl group which may have a substituent, a pyridyl group which may have a substituent, a pyrazolyl group which may have a substituent, a cyclohexyl group, a linear alkyl group having 1 to 5 carbon atoms which may have a substituent, a pyrazolylamino group which may have a substituent, or a phenylamino group which may have a substituent. R 2a represents a hydrogen atom or a methyl group. 3a represents a phenyl group which may have a substituent.

[0116] R 1a R represents a piperidyl group, a cyclohexyl group, or a linear alkyl group having 1 to 5 carbon atoms, which may have a substituent. 2a represents a hydrogen atom or a methyl group.

[0117] In general formula (1a), R 1a The substituent of the piperidyl group represented by the following formula is preferably a trifluoromethyl-substituted pyridyl group.

[0118] In general formula (1a), R 1a The substituent of the pyridyl group represented by the following formula is preferably a difluorophenyloxy group.

[0119] In general formula (1a), R 1a The substituent of the pyrazolyl group represented by the following formula is preferably a trifluoromethyl-substituted phenyl group.

[0120] In general formula (1a), R 1a The substituent of the linear alkyl group having 1 to 5 carbon atoms represented by the formula (I) is preferably a carbonylamino or piperidyl group.

[0121] In general formula (1a), R 3a The phenyl group represented by the formula (I) is preferably substituted with three alkyl groups having 1 to 5 carbon atoms (preferably isopropyl groups).

[0122] The compound represented by the above formula (1a) can be obtained by the method described in the production examples below or a method similar thereto.

[0123] 3. Screening Method Furthermore, the present invention encompasses an invention relating to a method for screening anticancer agents. The screening method of the present invention uses cells or viruses and measures the RNA methylation inhibitory activity of a test substance.

[0124] The cells used are preferably cancer cells, and the type of cancer is not particularly limited. Specific examples include pharyngeal cancer (lip cancer, gum cancer, tongue cancer, oral cancer, floor of the mouth cancer, salivary gland cancer, etc.), digestive cancer (esophageal cancer, stomach cancer, appendix cancer, colon cancer, rectal cancer, etc.), lung cancer, liver cancer, hepatocellular carcinoma, bile duct cancer, bone cancer, articular cartilage cancer, malignant melanoma of the skin, squamous cell carcinoma, other skin cancers, mesothelioma, breast cancer, uterine cancer (cervical cancer, endometrial cancer, etc.), ovarian cancer, prostate cancer, bladder cancer, brain tumor, thyroid cancer, non-Hodgkin's lymphoma, lymphocytic leukemia, sarcoma, and cancers of metastatic tissues originating from these as primary foci.

[0125] Specific cancer cells are not particularly limited and may be any known cancer cells of the above-mentioned cancer types. Of these, it is preferable to use cancer stem cells.

[0126] As the test substance, a low molecular weight compound is preferably used, and the methylation inhibitory effect on the above-mentioned cells or viruses is measured to select test substances having an inhibitory effect equal to or greater than a predetermined level.

[0127] In one preferred embodiment of the screening method of the present invention, the activity of an RNA methylation inhibitor is measured by measuring the activity of an enzyme that modifies RNA methylation, i.e., an RNA methyltransferase. A wide range of known RNA methyltransferases can be used. Specific examples include enzymes belonging to the ALKBH family and the Mettle family.

[0128] Among FTSJs, FTSJ1 in particular is thought to be a poor prognostic factor for cancer. Therefore, in screening anticancer drugs, it is preferable to measure the FTSJ1 inhibitory activity of test substances.

[0129] The specific method for measuring RNA methyltransferase activity is not particularly limited, and a wide variety of known methods can be used. Examples include ELISA, RIA, immunoprecipitation, bisulfite assay, quantitative PCR, and reporter assay. Among these, it is preferable to use the reporter assay because it allows for simple and accurate measurement.

[0130] In mammalian cells, FTSJ1 is predicted to 2'-O-methylate the 32nd and 34th nucleotides of tRNAs corresponding to the codons polyglutamine (Q), phenylalanine (F), methionine (M), and asparagine (N).

[0131] In a reporter assay, the sequence of the translation regulatory region to which the reporter region is bound preferably contains a sequence formed by binding at least one selected from the group consisting of glutamine, phenylalanine, tryptophan, methionine, and leucine.

[0132] The translation regulatory region is preferably a repeat sequence of the five amino acids mentioned above, i.e., polyglutamine, polyphenylalanine, polytryptophan, polymethionine, or polyleucine, but may also be a sequence containing a mixture of the five amino acids. Furthermore, other amino acids may also be included to the extent that they do not inhibit the function of the region.

[0133] The number of amino acid repeats is preferably 3 or more, more preferably 5 or more, and even more preferably 8 or more. The upper limit of the number of repeats is not particularly limited and can be set to 50, for example.

[0134] More specifically, examples of such translation regulatory regions include the sequences shown in any of SEQ ID NOS: 1 to 12. Examples of polyglutamine include SEQ ID NOS: 1 or 2, polyphenylalanine include SEQ ID NOS: 3 or 4, polytryptophan include SEQ ID NOS: 5, polymethionine include SEQ ID NOS: 6, and polyleucine include SEQ ID NOS: 7 to 12.

[0135] Furthermore, as a result of the research of the present inventors, it is predicted that YAP / TAZ is a protein that controls cancer stemness and whose protein synthesis is promoted by FTSJ1. Therefore, in addition to the reporter assay in which the above-mentioned translation regulatory region is added to the reporter, it is also preferable to perform a reporter assay in which expression is regulated by a transcription factor binding region (GTIIC) that contains 3 to 15 repeats of the sequence shown in SEQ ID NO: 13 (however, other sequences may be included between each repeat unit). In addition, it is also preferable to perform a sphere formation assay and / or mass spectrometry.

[0136] The reporter gene used in the reporter assay can be a wide variety of known reporter genes and is not particularly limited. Specific examples include the β-galactosidase gene, the chloramphenicol acetyltransferase gene derived from a bacterial transposon, and the firefly luciferase gene. Among these, the firefly luciferase gene is preferably used because it provides excellent detection sensitivity.

[0137] The method for linking the transcriptional regulatory region and the reporter gene can be a wide variety of known methods and is not particularly limited. Specifically, a method can be used in which the purified transcriptional sequence region is cleaved with an appropriate restriction enzyme and then linked to the reporter gene.

[0138] As the vector into which the ligated sequence is inserted, a wide range of known reporter assay vectors, such as plasmids, shuttle vectors, and helper plasmids, can be used.

[0139] The method for introducing a vector into a cell is not particularly limited, and a wide variety of known methods can be used, including, for example, electroporation, the spheroplast method, and the lithium acetate method.

[0140] The luminescence intensity of the cells into which the vector has been introduced is measured using a luminometer or the like in accordance with standard methods. In the screening method of the present invention, it is preferable to also measure the luminescence of a solvent such as DMSO as a control (blank), and calculate the assay value of the test substance as a percentage (%) of this.

[0141] In a reporter assay in which a translational regulatory region containing a sequence formed by the binding of at least one selected from the group consisting of glutamine, phenylalanine, tryptophan, methionine, and leucine is added to a reporter, the assay values ​​obtained are preferably selected by screening to be 100 or less, more preferably 80 or less, and even more preferably 40 or less.

[0142] Furthermore, in a reporter assay in which the transcriptional regulatory region (GTIIC) represented by SEQ ID NO: 13 is added to a reporter, assay values ​​obtained are preferably selected by screening to be 100 or less, more preferably 80 or less, and even more preferably 40 or less.

[0143] Another embodiment is a screening method comprising the steps of adding a methyl group donor to the above-described test substance to obtain a reaction product, and measuring the test substance using the reaction product.

[0144] As the methyl group donor, for example, one that can become a precursor of ATP, described below, by removing a methyl group from the methyl group donor is preferred, and a specific example is S-adenosylmethionine (hereinafter simply referred to as "SAM").

[0145] In the step of measuring the inhibitory effect of RNA methylation on cells or viruses, for example, ELISA, RIA, immunoprecipitation, bisulfite assay, quantitative PCR, reporter assay, and luciferase assay can be used.

[0146] In particular, when SAM is used as the methyl group donor in the step of adding the methyl group donor to the test substance, FTSJ1 contained in the test substance converts SAM to S-adenosylhomocysteine ​​(hereinafter simply referred to as "SAH"). The resulting SAH is then reacted with a reagent that converts it to adenosine diphosphate (hereinafter simply referred to as "ADP"), and a specific reagent is further added to the ADP to obtain adenosine triphosphate (hereinafter simply referred to as "ATP"). By combining this with an assay system such as a luciferase assay, the activity of FTSJ1 in the test substance can be evaluated more directly, and highly accurate evaluation results can be obtained.

[0147] 4. Method for predicting the efficacy or prognosis of FTSJ1 inhibitors against cancer The present invention also includes an invention relating to a method for predicting the efficacy of an FTSJ1 inhibitor against cancer, and a method for predicting cancer prognosis using an FSTJ1 inhibitor. As used herein, "prognosis" is defined as the medical outlook for a patient after chemotherapy.

[0148] The above-mentioned efficacy of an FTSJ1 inhibitor refers to the effect of an FTSJ1 inhibitor on cancer pathology. In other words, the method of predicting the efficacy of an FTSJ1 inhibitor of the present invention encompasses the concept of a method of predicting the sensitivity of a cancer patient, or cancer tissue or cancer cells collected from a cancer patient, to an FTSJ1 inhibitor. Furthermore, the method of predicting the efficacy of an FTSJ1 inhibitor of the present invention also encompasses the concept of a method of predicting the resistance of a cancer patient, or cancer tissue or cancer cells collected from a cancer patient, to an FTSJ1 inhibitor.

[0149] The method of the present invention for predicting the efficacy of an FTSJ1 inhibitor against cancer and the method of predicting the prognosis of cancer using an FTSJ1 inhibitor include a step A of measuring the expression level of FTSJ1 in a sample.

[0150] Here, cancer tissues or cancer cells derived from living organisms (including humans and animals) can be suitably used as specimens. Specifically, cancer tissues or cancer cells collected from patients (cancer patients) can be used.

[0151] The expression level of FTSJ1 can be measured by a wide range of known methods without any particular limitation, and either an immunological method or a genetic method can be suitably used.

[0152] The immunological technique is not particularly limited, and examples thereof include ELISA, immunostaining, flow cytometry, and immunoblot.

[0153] The genetic techniques are not particularly limited, and examples include Western blotting and RT-PCR.

[0154] The method of predicting the effectiveness of an FTSJ1 inhibitor against cancer and the method of predicting the prognosis of cancer using an FTSJ1 inhibitor of the present invention may further include, after the above step A, step B for determining the effectiveness of the FTSJ1 inhibitor against cancer or for determining the prognosis of the patient's cancer pathology based on the FTSJ1 expression level obtained in step A.

[0155] In particular, in step B, it is preferable to set a predetermined cutoff value for the expression level of FTSJ1 in the sample obtained in step A. Then, for example, when predicting the effectiveness of an FTSJ1 inhibitor, the effectiveness of FTSJ1 is predicted to be high for samples obtained in step A with an expression level equal to or higher than the cutoff value, and conversely, the effectiveness of FTSJ1 is predicted to be low for samples with an expression level lower than the cutoff value.

[0156] Similarly, in a method for predicting the pathological condition of a patient's cancer after use of an FTSJ1 inhibitor, for example, for a sample in which an expression level equal to or greater than a predetermined cutoff value is obtained in step A, it is predicted that the patient will have a good prognosis after use of an FTSJ1 inhibitor, and conversely, for a sample in which an expression level equal to or less than the cutoff value is obtained, it is predicted that the patient will have a poor prognosis after use of an FTSJ1 inhibitor.

[0157] Furthermore, the method of predicting the efficacy of an FTSJ1 inhibitor against cancer and the method of predicting cancer prognosis using an FTSJ1 inhibitor of the present invention can be widely applied to known cancer types without any particular limitation, specifically, glioblastoma (malignant brain tumor), pancreatic cancer, acute myeloid leukemia, lung cancer, liver cancer, kidney cancer, stomach cancer, and breast cancer.

[0158] 5. Anticancer drug efficacy assessment marker The present invention also includes an invention relating to a marker for determining the effectiveness of an anticancer drug. The marker is a genetic marker, and is a genetic marker associated with sensitivity to an FTSJ1 inhibitor and a genetic marker associated with resistance to an FTSJ1 inhibitor.

[0159] These markers are used to determine the sensitivity of a patient to an FTSJ1 inhibitor based on whether they are detected in a sample (tissue, cells, etc.) collected from the patient (including humans and animals). If a gene marker associated with FTSJ1 inhibitor sensitivity is detected in the sample, it is presumed that an FTSJ1 inhibitor will be effective during chemotherapy for that patient. On the other hand, if a gene marker associated with FTSJ1 inhibitor resistance is detected in the sample, it is presumed that an FTSJ1 inhibitor will not be effective during chemotherapy for that patient.

[0160] These FTSJ1-related markers for determining the efficacy of anticancer drugs (gene markers associated with sensitivity to FTSJ1 inhibitors or gene markers associated with resistance to FTSJ1 inhibitors) are preferably FTSJ1-modified nucleic acid RNAs.

[0161] Furthermore, the FTSJ1 inhibitor resistance-associated gene marker is preferably at least one selected from the group consisting of AHNAK2 (SEQ ID NO: 14), ESYT1 (SEQ ID NO: 15), SRGAP1 (SEQ ID NO: 16), RHOF (SEQ ID NO: 17), MIR4746 (SEQ ID NO: 18), UBXN6 (SEQ ID NO: 19), COX16 (SEQ ID NO: 20), FTH1 (SEQ ID NO: 21), LPAR1 (SEQ ID NO: 22), ANKRD29 (SEQ ID NO: 23), TWIST2 (SEQ ID NO: 24), JKAMP (SEQ ID NO: 25), PRKAA2 (SEQ ID NO: 26), CSTF2T (SEQ ID NO: 27), THSD4 (SEQ ID NO: 28), MAGI1 (SEQ ID NO: 29), UBE2L3 (SEQ ID NO: 30), GPLD1 (SEQ ID NO: 31), FRYL (SEQ ID NO: 32), and MYO9A (SEQ ID NO: 33).

[0162] The gene markers associated with FTSJ1 inhibitor sensitivity are RBM15 (SEQ ID NO: 34), NASP (SEQ ID NO: 35), PRPF38A (SEQ ID NO: 36), C1orf50 (SEQ ID NO: 37), PEX16 (SEQ ID NO: 38), ZNF213 (SEQ ID NO: 39), FEM1B (SEQ ID NO: 40), RFXAP (SEQ ID NO: 41), SAP18 (SEQ ID NO: 42), AARS2 (SEQ ID NO: 43), RCC2 (SEQ ID NO: 44), YARS1 (SEQ ID NO: 45), RBM It is preferable that the gene is at least one selected from the group consisting of: 10 (SEQ ID NO: 46), RPL5 (SEQ ID NO: 47), ZNHIT2 (SEQ ID NO: 48), OSGIN2 (SEQ ID NO: 49), EGLN3 (SEQ ID NO: 50), TRPTI (SEQ ID NO: 51), CRACDL (SEQ ID NO: 52), CAPG (SEQ ID NO: 53), RAB11FIP3 (SEQ ID NO: 54), CALHM5 (SEQ ID NO: 55), BICD1 (SEQ ID NO: 56) and FTSJ1 (SEQ ID NO: 57).

[0163] 6. Kit for predicting the efficacy of FTSJ1 inhibitors The present invention also encompasses a kit containing the above-mentioned marker for determining the effectiveness of an anticancer drug. When at least one of the above-mentioned gene markers associated with resistance to an FTSJ1 inhibitor is detected in a sample, it is determined that an FTSJ1 inhibitor is ineffective in chemotherapy for the patient.

[0164] On the other hand, when at least one of the above-mentioned gene markers associated with sensitivity to an FTSJ1 inhibitor is detected in a sample, it is determined that an FTSJ1 inhibitor is effective in chemotherapy for the patient.

[0165] Such a kit is not particularly limited as long as it utilizes a mechanism for detecting the above-mentioned genetic markers in a sample. For example, a kit may be used in which cDNA is obtained from a sample and amplified by PCR to detect the genetic markers. In this case, the kit of the present invention preferably includes, for example, primers for each genetic marker for performing PCR.

[0166] Although the embodiments of the present invention have been described above, the present invention is not limited to these examples, and it goes without saying that the present invention can be embodied in various forms without departing from the spirit of the present invention. [Example]

[0167] Hereinafter, the embodiments of the present invention will be described in more detail based on examples, but the present invention is not limited to these examples.

[0168] (Experimental example) screening The human gastric cancer cell line NUGC3 was cultured in DMEM medium (high glucose, L-glutamine, and phenol red, Fujifilm Wako Pure Chemical Industries, Ltd.) containing 10% fetal bovine serum (Thermo Fisher Scientific) and penicillin / streptomycin (Fujifilm Wako Pure Chemical Industries, Ltd.) (hereinafter, this medium will be referred to simply as "DMEM + 10% FBS + 1x P / S"). NUGC3 cells were then seeded onto 60 mm culture dishes (BioLite) and allowed to reach 80% confluence after 24 hours. Next, 5 μg of reporter plasmid and 15 μL of lipofection reagent (TransIT-LT1, Mirus) were mixed in 500 μL of Opti-MEM (Thermo Fisher Scientific) medium. The resulting complex was added to the medium of NUGC3 cells cultured in 60 mm culture dishes. For reporter plasmids, 5 μg of polyglutamine luciferase reporter was transfected (polyglutamine luciferase reporter has an IRES-expressed Renilla luciferase downstream of polyglutamine-tagged firefly luciferase as an internal reference). For YAP / TAZ activity reporter, 3 μg of 8xGTIIC plasmid was transfected simultaneously with 2 μg of Renilla luciferase reporter as an internal reference. After 24 hours, cells were detached with 0.05 w / v% trypsin-0.53 mmol / L EDTA·4Na solution (Fujifilm Wako Pure Chemical Industries, Ltd.) and re-seeded onto 96-well plates (BioLite) to reach 80% confluence after 24 hours. Twenty-four hours after seeding the cells back onto the 96-well plate, the medium in each well was replaced with 100 μL of medium (DMEM + 10% FBS + antibiotics) containing 10 μM or 5 μM of the compound to be evaluated. After another 24 hours, the compound-containing medium was removed, and the 96-well plate was transferred to ice. After washing with 100 μL of phosphate-buffered saline (PBS, Fujifilm Wako Pure Chemical Industries, Ltd.), 20 μL of Passive Lysis Buffer (1X) included in the Dual-Luciferase Reporter Assay System (Promega) was added. The cells were then gently rocked at room temperature for 15 minutes to lyse the cells.After confirming cell lysis, 10 μL of the lysate from each well was transferred to the corresponding well of a white 96-well plate (Greiner). Firefly luciferase luminescence was induced by adding 100 μL of a mixture of Luciferase Assay Buffer II and Luciferase Assay Substrate from the Dual-Luciferase Reporter Assay System (Promega) to each well of the white 96-well plate, and the luminescence intensity was measured using a microluminometer (Berthold). Subsequently, 100 μL of a mixture of Stop & Glo Buffer and Stop & Glo Substrate from the Dual-Luciferase Reporter Assay System (Promega) was added to quench the firefly luciferase luminescence and induce Renilla luciferase luminescence. The luminescence intensity was similarly measured using a microluminometer (Berthold). The ratio of the luminescence intensity of firefly luciferase to that of Renilla luciferase was calculated, and the luminescence intensity ratio of the well to which each compound was added was calculated as a percentage, with the luminescence intensity ratio of the well to which DMSO (dimethyl sulfoxide, used as a solvent for compound solutions) was added being set at 100%. The inhibition results for each compound are shown in Table 1 below. Note that, unless otherwise specified, the measured values ​​in the table indicate the results of evaluation after adding 10 μM of the compound to be evaluated.

[0169] [Table 1] JPEG2025081579000008.jpg254170JPEG2025081579000009.jpg242170JPEG2025081579000010.jpg246170JPEG2025081579000011.jpg226170JPEG2025081579000012.jpg240170JPEG2025081579000013.jpg255170JPEG2025081579000014.jpg252170JPEG2025081579000015.jpg254169JPEG2025081579000016.jpg244170JPEG2025081579000017.jpg240169JPEG2025081579000018.jpg248170JPEG2025081579000019.jpg223169JPEG2025081579000020.jpg245169JPEG2025081579000021.jpg249169JPEG2025081579000022.jpg222170JPEG2025081579000023.jpg233170JPEG2025081579000024.jpg255169JPEG2025081579000025.jpg247170JPEG2025081579000026.jpg215169JPEG2025081579000027.jpg253169JPEG2025081579000028.jpg233170JPEG2025081579000029.jpg223169JPEG2025081579000030.jpg253170JPEG2025081579000031.jpg255169JPEG2025081579000032.jpg248170JPEG2025081579000033.jpg231169JPEG2025081579000034.jpg223169JPEG2025081579000035.jpg223169JPEG2025081579000036.jpg253170JPEG2025081579000037.jpg91170JPEG2025081579000038.jpg255164JPEG2025081579000039.jpg243167JPEG2025081579000040.jpg235167JPEG2025081579000041.jpg255163JPEG2025081579000042.jpg224168JPEG2025081579000043.jpg245167JPEG2025081579000044.jpg255165JPEG2025081579000045.jpg236167JPEG2025081579000046.jpg251168JPEG2025081579000047.jpg255165JPEG2025081579000048.jpg220166JPEG2025081579000049.jpg224167JPEG2025081579000050.jpg234167JPEG2025081579000051.jpg251167JPEG2025081579000052.jpg245167JPEG2025081579000053.jpg235167JPEG2025081579000054.jpg241167JPEG2025081579000055.jpg251167JPEG2025081579000056.jpg217167JPEG2025081579000057.jpg230167JPEG2025081579000058.jpg236167JPEG2025081579000059.jpg232167JPEG2025081579000060.jpg246167JPEG2025081579000061.jpg255165JPEG2025081579000062.jpg238167JPEG2025081579000063.jpg245167JPEG2025081579000064.jpg255164JPEG2025081579000065.jpg254166JPEG2025081579000066.jpg238167JPEG2025081579000067.jpg227167JPEG2025081579000068.jpg255164JPEG2025081579000069.jpg234166JPEG2025081579000070.jpg255164JPEG2025081579000071.jpg239167JPEG2025081579000072.jpg245166JPEG2025081579000073.jpg222167JPEG2025081579000074.jpg239167JPEG2025081579000075.jpg221167JPEG2025081579000076.jpg244167JPEG2025081579000077.jpg236167JPEG2025081579000078.jpg230167JPEG2025081579000079.jpg225167JPEG2025081579000080.jpg224167JPEG2025081579000081.jpg244167JPEG2025081579000082.jpg219167JPEG2025081579000083.jpg255167JPEG2025081579000084.jpg233166JPEG2025081579000085.jpg246167JPEG2025081579000086.jpg251167JPEG2025081579000087.jpg255167JPEG2025081579000088.jpg255165JPEG2025081579000089.jpg235167JPEG2025081579000090.jpg216167JPEG2025081579000091.jpg255167JPEG2025081579000092.jpg225167JPEG2025081579000093.jpg245167JPEG2025081579000094.jpg232167JPEG2025081579000095.jpg231167JPEG2025081579000096.jpg230167JPEG2025081579000097.jpg255164JPEG2025081579000098.jpg228167JPEG2025081579000099.jpg236167JPEG2025081579000100.jpg222167JPEG2025081579000101.jpg234167JPEG2025081579000102.jpg242167JPEG2025081579000103.jpg228167JPEG2025081579000104.jpg223168JPEG2025081579000105.jpg231168JPEG2025081579000106.jpg242167JPEG2025081579000107.jpg232167JPEG2025081579000108.jpg240167JPEG2025081579000109.jpg234167JPEG2025081579000110.jpg96167.

[0170] Screening Tests Human embryonic kidney (HEK293) cells constitutively expressing FLAG-tagged FTSJ1 were engineered. FLAG-FTSJ1 was isolated from the cell lysate using anti-FLAG M2 antibody affinity gel (Sigma-Aldrich, cat# A2220-10ML) and eluted with FLAG peptide (Sigma-Aldrich, cat# F3290-25MG). The protein concentration of the eluate was measured, and then serially diluted from 0 to 20 ng / reaction to obtain 12 different enzyme dilutions. Total RNA was also extracted from HEK293-FTSJ1-KO cells, in which FTSJ1 was knocked out, using TRIzol Reagent (ThermoFisher, cat# 15596018). To measure the activity of FLAG-FTSJ1 when it methylates RNA, we used the enzyme dilutions described above, total RNA (1,000 ng / reaction), and a methyltransferase activity assay kit (Promega, cat. #V7601). The conversion of S-adenosyl methionine (SAM) to S-adenosyl homocysteine ​​(SAH) produced by methylation of 1,000 ng / reaction of total RNA by FLAG-FTSJ1 in each dilution was measured as luciferase luminescence using the MTase-Glo Reagent included in the assay kit. The luminescence value at 0 ng of FLAG-FTSJ1 was used as the background value, and a graph was created by subtracting the background value from the luminescence values ​​at each dilution. As shown in Figure 3A, a dose-dependent increase in luminescence was observed, confirming the detectable enzymatic activity of FTSJ1.

[0171] Using the above system, we confirmed the inhibitory effect of PVZF2001, an FTSJ1 inhibitor, on the methyltransferase reaction. Using 10 ng / reaction of FLAG-FTSJ1, 1,000 ng / reaction of total RNA, and a methyltransferase activity assay kit, we measured the luminescence intensity when different concentrations of PVZF2001 were mixed. As shown in Figure 3B, we observed a concentration-dependent decrease in luminescence intensity, confirming that PVZF2001 inhibits the methyltransferase reaction catalyzed by FTSJ1.

[0172] Antitumor effect evaluation test A suspension of breast cancer triple-negative cell line MDA-MB-231 cells suspended in phosphate buffered saline (PBS, Fujifilm Wako Pure Chemical Industries) was injected subcutaneously into the lumbar region of immunodeficient mice (BALB / c-nu / nu, female, 6 weeks old, purchased from Shimizu Laboratory Materials) at a dose of 1 × 10 using a syringe with a 23G needle (Terumo Corporation). 6 The cancer cells were inoculated at 100 μL per cell. The cancer cells were subcutaneously implanted in the mice to form tumor tissue, and the tumor volume reached 100 mm. 3 After confirming that the tumor volume reached the target value, the obtained tumor model mice were used in the following evaluation test. First, the compound to be evaluated was dissolved in corn oil (Sigma Aldrich) and administered intraperitoneally. PVZF0024 was administered at 100 mg / kg, and PVZF2001 was administered at 20 mg / kg to the tumor model mice every other day. After each administration, the tumor diameter was measured with a vernier caliper. The tumor volume was calculated as V = (3.14 × D × d 2 ) / 6 (where V is tumor volume, D is tumor major axis, and d is tumor minor axis) (Wu. et al., Clin. Cancer Res., 2013 Oct 15;19(20):5699-5710).

[0173] The antitumor effects of PVZF0024 and PVZF2001 are shown in Figures 1 and 2. As shown in each figure, both compounds were confirmed to significantly suppress tumor growth, and no weight loss was observed due to administration.

[0174] A study to investigate genetic markers related to sensitivity to FTSJ1 inhibitors To identify genes that influence sensitivity and resistance to FTSJ1 inhibitors, we analyzed a cell line panel (JFCR39) using PVZF2001. Thirty-nine human cancer cell lines were treated with PVZF2001 at concentrations of 0.01 μM, 0.1 μM, 1 μM, 10 μM, and 100 μM for 48 hours. Cell proliferation was measured colorimetrically with sulforhodamine B, and the Log GI50 values ​​shown in Figure 4A were obtained. The GI50 value is defined as the concentration at which the increase in cell number due to drug treatment is reduced by 50%, relative to the increase in cell number in the untreated sample (negative control) cultured for 48 hours before drug exposure. To extract the target gene group from this, a COMPARE analysis was first performed using gene expression and Log GI50 values ​​in all cell lines, identifying the gene group shown in SEQ ID NOs: 14 to 28 as FTSJ1 inhibitor resistance-associated gene markers and the gene group shown in SEQ ID NOs: 34 to 47 as FTSJ1 inhibitor sensitivity-associated gene markers. Furthermore, limited to malignant brain tumors and lung cancers, which are clearly differentiated in sensitivity and resistance to PVZF2001, gene expression data for highly sensitive cell lines (e.g., U251, SF-539, SNB-75, NCI-H522, DMS114) and highly resistant cell lines (e.g., SNB-78, NCI-H23) were used to analyze the gene group whose expression was commonly increased in each of the cell lines. As a result, the gene group shown in SEQ ID NOs: 29 to 33 was identified as FTSJ1 inhibitor resistance-associated gene markers, and the gene group shown in SEQ ID NOs: 48 to 56 was identified as FTSJ1 inhibitor sensitivity-associated gene markers. Using these genetic markers, we analyzed a database (TCGA) of gene expression information obtained from patient samples and found that patients with high FTSJ1 gene expression levels had high expression of gene markers associated with FTSJ1 inhibitor sensitivity, and conversely, patients with low FTSJ1 gene expression levels had high expression of gene markers associated with FTSJ1 inhibitor resistance. In other words, it was found that the FTSJ1 gene expression level itself is a gene that determines sensitivity and resistance to FTSJ1 inhibitors (SEQ ID NO: 57).

[0175] To verify whether expression analysis of these gene groups contributes to predicting the anticancer effects of FTSJ1 inhibitors, we actually analyzed the anticancer effects of the FTSJ1 inhibitor PVZF2001 on human malignant brain tumor cell lines, using the FTSJ1 gene as an example. Human malignant brain tumor cell lines MGG4, MGG8, MGG18, and MGG23 were treated with stem cell medium (Neurobasal, B-27, N-2, 20 ng / mL EGF, and 20 ng / mL bFGF) containing PVZF2001 at different concentrations (0 nM, 200 nM, 500 nM, 1000 nM, 2000 nM, and 5000 nM) for 1 week, and anchorage-independent cell growth was assessed by sphere formation. As shown in Figure 4B, MGG4, MGG8, and MGG18, which express high levels of FTSJ1, exhibited growth inhibition at low concentrations of PVZF2001. However, MGG23, which expresses low levels of FTSJ1, exhibited cell proliferation even at a high concentration of 2000 nM.

[0176] (Production Example 1) (E)-1-(2,3-Dihydrobenzo[b][1,4]dioxin-6-yl)-3-(2-hydroxyphenyl)prop-2-en-1-one (PVZF0024) 178 mg of 1-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)ethan-1-one and 122 mg of 2-hydroxybenzaldehyde were dissolved in 20 mL of ethanol. 0.5 mL of 40% sodium hydroxide solution was added and the mixture was stirred at 60°C for 10 hours. After neutralization with acetic acid, the mixture was extracted with chloroform and purified by silica gel chromatography to obtain (E)-1-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)-3-(2-hydroxyphenyl)prop-2-en-1-one. 141 mg of the above compound was dissolved in 10 mL of ethanol, 121 μL of hydrazine monohydrate was added, and the mixture was stirred at 80°C for 5 hours. The ethanol was evaporated under reduced pressure, and the resulting residue was purified by silica gel chromatography to obtain 31 mg of the desired compound in 31% yield. ESI (m / z): 297 (M+H) + 1H-NMR (400 MHz, DMSO-d6) δ 9.66 (d, J = 25.4 Hz, 1H), 7.25-7.27 (m, 1H), 7.05-7.12 (m, 3H), 6.74-6.86 (m, 3H), 4.95 (td, J = 10.5, 2.8 Hz, 1H), 4.25 (s, 4H), 3.32-3.38 (m, 1H), 2.66 (dd, J = 16.3, 10.5 Hz, 1H)

[0177] (Production Example 2) 2,4,6-Triisopropyl-N-(piperidin-4-yl)benzenesulfonamide (PVZF2005) 151 mg of 2,4,6-triisopropylbenzenesulfonyl chloride was dissolved in 10 mL of dichloromethane, and 50 mg of piperidin-4-amine and 120 μL of pyridine were added in an ice bath and stirred at room temperature for 4 hours. Methanol was added to terminate the reaction, and dichloromethane was evaporated under reduced pressure. The resulting residue was purified by silica gel chromatography to give 158 mg of the desired compound in 86% yield. ESI (m / z): 367 (M+H) + 1 H-NMR (400 MHz, DMSO-d6) δ 7.29-7.48 (s, 2H), 4.05 (td, J = 13.4, 6.7 Hz, 3H), 3.43 (m, 8H), 1.91-1.95 (m, 1H), 1.44 (td, J = 11.8, 3.1Hz, 1H), 1.05-1.38 (m, 18H)

[0178] (Production Example 3) 2,4,6-Triisopropyl-N-(piperidin-3-ylmethyl)benzenesulfonamide (PVZF2008) 151 mg of 2,4,6-triisopropylbenzenesulfonyl chloride was dissolved in 10 mL of dichloromethane, and 57 mg of piperidin-3-ylmethanamine and 120 μL of pyridine were added in an ice bath and stirred at room temperature for 4 hours. Methanol was added to terminate the reaction, and dichloromethane was evaporated under reduced pressure. The resulting residue was purified by silica gel chromatography to give 129 mg of the desired compound in 68% yield. ESI (m / z): 381 (M+H) + 1 H-NMR (400 MHz, DMSO-d6) δ 7.22-7.28 (m, 2H), 4.08 (td, J = 13.3, 6.7 Hz, 2H), 3.57-3.64 (m, 1H), 2.90-2.97 (m, 1H), 2.62-2.74 (m, 3H), 1.70-1.91 (m, 3H), 1.09-1.29 (m, 22H)

[0179] (Production Example 4) N-(2-(cyclohex-1-en-1-yl)ethyl)-[1,1'-biphenyl]-4-sulfonamide (PVZF2035) 126 mg of [1,1'-biphenyl]-4-sulfonyl chloride was dissolved in 10 mL of dichloromethane, and 62.5 mg of 2-(cyclohex-1-en-1-yl)ethan-1-amine and 120 μL of pyridine were added in an ice bath and stirred at room temperature for 4 hours. Methanol was added to terminate the reaction, and the dichloromethane was evaporated under reduced pressure. The resulting residue was purified by silica gel chromatography to give 123 mg of the desired compound in 72% yield. ESI (m / z): 342 (M+H) + 1 H-NMR (400 MHz, DMSO-d6) δ 7.85-7.90 (m, 4H), 7.74-7.76 (m, 2H), 7.64 (d, J = 23.2 Hz, 1H), 7.52 (t, J = 7.7 Hz, 2H), 7.42-7.46 (m, 1H), 5.33 (s, 1H), 2.83 (t, J = 7.4 Hz, 2H), 2.01 (t, J = 7.2 Hz, 2H), 1.89 (s, 2H), 1.80 (s, 2H), 1.43-1.56 (m, 4H)

[0180] (Production Example 5) N-(2,2,6,6-tetramethylpiperidin-4-yl)-[1,1'-biphenyl]-4-sulfonamide (PVZF0036) 126 mg of [1,1'-biphenyl]-4-sulfonyl chloride was dissolved in 10 mL of dichloromethane, and 78 mg of 2,2,6,6-tetramethylpiperidin-4-amine and 120 μL of pyridine were added in an ice bath and stirred at room temperature for 4 hours. Methanol was added to terminate the reaction, and dichloromethane was distilled off under reduced pressure. The resulting residue was purified by silica gel chromatography to obtain 153 mg of the target compound in an 82% yield. ESI (m / z): 373 (M+H) + 1 H-NMR (400 MHz, DMSO-d6) δ 7.88-7.92 (m, 4H), 7.75 (t, J = 8.4 Hz, 2H), 7.49-7.55 (m, 2H), 7.42-7.47 (m, 1H), 3.46-3.57 (m, 1H), 1.54-1.61 (m, 2H), 1.39 (m, 2H), 1.13-1.37 (m, 12H)

[0181] (Production Example 6) N-Cyclohexyl-4-(tert-pentyl)benzenesulfonamide (PVZF0039) 123 mg of 4-(tert-pentyl)benzenesulfonyl chloride was dissolved in 10 mL of dichloromethane, and 50 mg of cyclohexaneamine and 120 μL of pyridine were added in an ice bath and stirred at room temperature for 4 hours. Methanol was added to terminate the reaction, and dichloromethane was distilled off under reduced pressure. The resulting residue was purified by silica gel chromatography to obtain 105 mg of the target compound in a 68% yield. ESI (m / z): 310 (M+H) + 1H-NMR (400 MHz, DMSO-d6) δ 7.72-7.81 (m, 2H), 7.56 (s, 1H), 7.53 (d, J = 8.0 Hz, 2H), 2.91 (s, 1H), 1.63 (q, J = 7.4 Hz, 2H), 1.55(d,J = 4.6Hz, 2H), 1.41-1.44 (m, 1H), 1.27 (s, 6H), 1.05-1.15 (m, 4H), 1.01 (d, J = 11.0 Hz, 2H), 0.58 (t, J = 7.3 Hz, 3H)

[0182] (Production Example 7) 4-Cyclohexyl-N-pentylbenzenesulfonamide (PVZF2065) 129 mg of 4-cyclohexylbenzenesulfonyl chloride was dissolved in 10 mL of dichloromethane, and 44 mg of pentan-1-amine and 120 μL of pyridine were added in an ice bath. The mixture was stirred at room temperature for 4 hours. Methanol was added to terminate the reaction, and the dichloromethane was distilled off under reduced pressure. The resulting residue was purified by silica gel chromatography to give 121 mg of the target compound in 78% yield. ESI (m / z): 310 (M+H) + 1 H-NMR (400 MHz, DMSO-d6) δ 7.67-7.70 (m, 2H), 7.47 (s, 1H), 7.43 (d, J = 8.0 Hz, 2H), 2.57-2.72 (m, 3H), 1.79 (d, J = 10.7 Hz, 4H), 1.71 (d, J = 12.7 Hz, 1H), 1.25-1.56 (m, 7H), 1.12-1.23 (m, 4H), 0.79 (dd, J = 7.0, 6.0 Hz, 3H)

[0183] (Production Example 8) N,4-Dicyclohexylbenzenesulfonamide (PVZF2066) 129 mg of 4-cyclohexylbenzenesulfonyl chloride was dissolved in 10 mL of dichloromethane, and 50 mg of cyclohexanamine and 120 μL of pyridine were added in an ice bath. The mixture was stirred at room temperature for 4 hours. Methanol was added to terminate the reaction, and the dichloromethane was distilled off under reduced pressure. The resulting residue was purified by silica gel chromatography to obtain 135 mg of the target compound in 84% yield. ESI (m / z): 322 (M+H). + 1 H-NMR (400 MHz, DMSO-d6) δ 7.71 (d, J = 8.3 Hz, 2H), 7.56 (s, 1H), 7.42 (d, J = 8.3 Hz, 2H), 2.91 (s, 1H), 2.56-2.67 (m, 1H), 1.78-1.85 (m, 4H), 1.71 (d, J = 12.4 Hz, 1H), 1.56 (d, J = 6.6 Hz, 4H), 1.32-1.43 (m, 5H), 1.25 (t, J = 12.2 Hz, 1H), 1.00-1.16 (m, 5H)

[0184] (Production Example 9) 4-Cyclohexyl-N-(cyclohexylmethyl)benzenesulfonamide (PVZF2069) 129 mg of 4-cyclohexylbenzenesulfonyl chloride was dissolved in 10 mL of dichloromethane, and 58 mg of cyclohexylmethanamine and 120 μL of pyridine were added in an ice bath and stirred at room temperature for 4 hours. Methanol was added to terminate the reaction, and the dichloromethane was evaporated under reduced pressure. The resulting residue was purified by silica gel chromatography to give 131 mg of the desired compound in 78% yield. ESI (m / z): 336 (M+H) + 1H-NMR (400 MHz, DMSO-d6) δ 7.68 (d, J = 8.3 Hz, 2H), 7.49 (s, 1H), 7.43 (d, J = 8.3 Hz, 2H), 2.53-2.67 (m, 6H), 1.77-1.84 (m, 3H), 1.71 (d, J = 12.4 Hz, 2H), 1.61 (d, J = 11.2 Hz, 3H), 1.35-1.46 (m, 2H), 1.22-1.32 (m, 1H), 1.07-1.15 (m, 3H), 0.74-0.82 (m, 2H)

[0185] (Production Example 10) 4-Bromo-2-isopropyl-N-(2,2,6,6-tetramethylpiperidin-4-yl)benzenesulfonamide (PVZF2074) 148 mg of 4-bromo-2-isopropylbenzenesulfonyl chloride was dissolved in 10 mL of dichloromethane, and 78 mg of 2,2,6,6-tetramethylpiperidin-4-amine and 120 μL of pyridine were added in an ice bath and stirred at room temperature for 4 hours. Methanol was added to terminate the reaction, and the dichloromethane was evaporated under reduced pressure. The resulting residue was purified by silica gel chromatography to give 175 mg of the desired compound in 85% yield. ESI (m / z): 417 (M+H) + 1 H-NMR (400 MHz, DMSO-d6) δ 8.14 (bs, 1H), 7.78 (d, J = 8.5 Hz, 2H), 7.57-7.60 (m, 1H), 3.75-3.82 (m, 1H), 1.60 (s, 2H), 1.13-1.39 (m, 22H)

[0186] (Production Example 11) 2-(tert-butyl)-N-(2,2,6,6-tetramethylpiperidin-4-yl)benzenesulfonamide (PVZF2075) 116 mg of 2-(tert-butyl)benzenesulfonyl chloride was dissolved in 10 mL of dichloromethane, and 78 mg of 2,2,6,6-tetramethylpiperidin-4-amine and 120 μL of pyridine were added in an ice bath and stirred at room temperature for 4 hours. Methanol was added to terminate the reaction, and dichloromethane was distilled off under reduced pressure. The resulting residue was purified by silica gel chromatography to obtain 141 mg of the target compound PVZF2075 in an 80% yield. ESI (m / z): 353 (M+H)+ 1 H-NMR (400 MHz, DMSO-d6) δ 8.05 (d, J = 8.0 Hz, 2H), 7.66 (d, J = 8.0 Hz, 1H), 7.53-7.56 (m, 1H), 7.43 (t, J = 7.7 Hz, 1H), 1.68-1.75 (m, 2H), 1.52 (s, 9H), 1.15-1.29 (m, 16H)

[0187] (Production Example 12) 2-Isopropyl-4-methoxy-5-methyl-N-(2,2,6,6-tetramethylpiperidin-4-yl)benzenesulfonamide (PVZF2076) 131 mg of 2-isopropyl-4-methoxy-5-methylbenzenesulfonyl chloride was dissolved in 10 mL of dichloromethane, and 78 mg of 2,2,6,6-tetramethylpiperidin-4-amine and 120 μL of pyridine were added in an ice bath and stirred at room temperature for 4 hours. Methanol was added to terminate the reaction, and the dichloromethane was evaporated under reduced pressure. The resulting residue was purified by silica gel chromatography to give 145 mg of the desired compound in 76% yield. ESI (m / z): 383 (M+H) + 1 H-NMR (400 MHz, DMSO-d6) δ 7.77 (bs, 1H), 7.64 (s, 1), 7.03 (s, 1H), 3.88 (s, 3H), 3.76-3.83 (m, 1H), 2.20-2.08 (3H), 1.59 (d, J = 12.4Hz, 2H), 1.13-1.33 (m, 22H)

[0188] (Manufacturing Example 13) 2,5-Diisopropyl-N-(2,2,6,6-tetramethylpiperidin-4-yl)benzenesulfonamide (PVZF2077) 130 mg of 2,5-diisopropylbenzenesulfonyl chloride was dissolved in 10 mL of dichloromethane, and 78 mg of 2,2,6,6-tetramethylpiperidin-4-amine and 120 μL of pyridine were added in an ice bath and stirred at room temperature for 4 hours. Methanol was added to terminate the reaction, and dichloromethane was distilled off under reduced pressure. The resulting residue was purified by silica gel chromatography to obtain 164 mg of the target compound in an 86% yield. ESI (m / z): 381 (M+H)+ 1 H-NMR (400 MHz, DMSO-d6) δ 7.93 (bs, 1H), 7.71-7.79 (m, 1H), 7.44-7.54 (m, 2H), 3.76-3.83 (m, 1H), 3.54 (d, J = 25.6 Hz, 1H), 3.05-2.86 (1H), 1.54 (d, J = 11.5 Hz, 2H), 1.03-1.39 (m, 26H)

[0189] (Manufacturing Example 14) 2-Cyclopropyl-N-(2,2,6,6-tetramethylpiperidin-4-yl)benzenesulfonamide (PVZF2078) 108 mg of 2-cyclopropylbenzenesulfonyl chloride was dissolved in 10 mL of dichloromethane, and 78 mg of 2,2,6,6-tetramethylpiperidin-4-amine and 120 μL of pyridine were added in an ice bath and stirred at room temperature for 4 hours. Methanol was added to terminate the reaction, and dichloromethane was distilled off under reduced pressure. The resulting residue was purified by silica gel chromatography to obtain 133 mg of the target compound in a 79% yield. ESI (m / z): 337 (M+H)+ 1H-NMR (400 MHz, DMSO-d6) δ 8.02-7.91 (1H), 7.89 (d, J = 7.8 Hz, 1H), 7.52 (t, J = 7.4 Hz, 1H), 7.32 (t, J = 7.7 Hz, 1H), 7.05 (d, J = 7.8 Hz, 1H), 3.51 (s, 2H), 2.64-2.70 (m, 1H), 1.60 (d, J = 10.0 Hz, 2H), 1.03-1.35 (m, 16H), 0.80-0.84 (m, 2H)

[0190] (Manufacturing Example 15) 5-chloro-2-cyclopropyl-N-(2,2,6,6-tetramethylpiperidin-4-yl)benzenesulfonamide (PVZF2079) 125 mg of 5-chloro-2-cyclopropylbenzenesulfonyl chloride was dissolved in 10 mL of dichloromethane, and 78 mg of 2,2,6,6-tetramethylpiperidin-4-amine and 120 μL of pyridine were added in an ice bath and stirred at room temperature for 4 hours. Methanol was added to terminate the reaction, and dichloromethane was distilled off under reduced pressure. The resulting residue was purified by silica gel chromatography to obtain 152 mg of the target compound in an 82% yield. ESI (m / z): 371 (M+H)+ 1 H-NMR (400 MHz, DMSO-d6) δ 8.14 (d, J = 28.8 Hz, 1H), 7.86 (d, J = 1.2 Hz, 1H), 7.59 (d, J = 8.5 Hz, 1H), 7.08 (d, J = 8.5 Hz, 1H), 2.60-2.68 (m, 1H), 1.72-1.47 (2H), 1.44-1.04 (18H), 0.91-0.74 (bs, 2H)

[0191] (Manufacturing Example 16) N-Benzhydryl-2,4,6-triisopropylbenzenesulfonamide (PVZF2082)151 mg of 2,4,6-triisopropylbenzenesulfonyl chloride was dissolved in 10 mL of dichloromethane, and 92 mg of diphenylmethanamine and 120 μL of pyridine were added in an ice bath and stirred at room temperature for 4 hours. Methanol was added to terminate the reaction, and dichloromethane was distilled off under reduced pressure. The resulting residue was purified by silica gel chromatography to obtain 101 mg of the target compound in a 45% yield. ESI (m / z): 450 (M+H)+ 1 H-NMR (400 MHz, DMSO-d6) δ 8.73 (d, J = 9.3 Hz, 1H), 7.15-7.35 (m, 10H), 7.05-7.11 (m, 2H), 5.46 (d, J = 9.3 Hz, 1H), 4.07-4.13 (m, 2H), 2.84-2.92 (m, 1H), 1.14-1.24 (m, 6H), 1.05 (m, 12H)

[0192] (Manufacturing Example 17) 2,4,6-triisopropyl-N-(1,2,2,6,6-pentamethylpiperidin-4-yl)benzenesulfonamide (PVZF2085) 151 mg of 2,4,6-triisopropylbenzenesulfonyl chloride was dissolved in 10 mL of dichloromethane, and 85 mg of 1,2,2,6,6-pentamethylpiperidin-4-amine and 120 μL of pyridine were added in an ice bath and stirred at room temperature for 4 hours. Methanol was added to terminate the reaction, and dichloromethane was evaporated under reduced pressure. The resulting residue was purified by silica gel chromatography to give 118 mg of the desired compound in 54% yield. ESI (m / z): 437 (M+H) + 1 H-NMR (400 MHz, DMSO-d6) δ 7.23 (s, 2H), 3.94-4.08 (m, 3H), 2.89-2.96 (m, 1H), 2.60-2.62 (m, 3H), 1.29-1.41 (m, 5H), 1.20 (d, J = 6.8Hz, 18H), 0.96-1.06 (m, 12H)

[0193] (Manufacturing Example 18) 2,4,6-triisopropyl-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4-yl)benzenesulfonamide (PVZF2086) 151 mg of 2,4,6-triisopropylbenzenesulfonyl chloride was dissolved in 10 mL of dichloromethane, and 85 mg of N,2,2,6,6-pentamethylpiperidin-4-amine and 120 μL of pyridine were added in an ice bath and stirred at room temperature for 4 hours. Methanol was added to terminate the reaction, and dichloromethane was distilled off under reduced pressure. The resulting residue was purified by silica gel chromatography to obtain 50 mg of the target compound in a 23% yield. ESI (m / z): 437 (M+H)+ 1 H-NMR (400 MHz, DMSO-d6) δ 7.27 (s, 2H), 4.02 (td, J = 13.4, 6.7 Hz, 3H), 2.93 (td, J = 13.6, 6.7 Hz, 1H), 2.64 (s, 3H), 1.49-1.63 (m, 3H)1.04-1.37 (m, 32H)

[0194] (Manufacturing Example 19) N-(2,6-dimethylpiperidin-4-yl)-2,4,6-triisopropylbenzenesulfonamide (PVZF2132) 151 mg of 2,4,6-triisopropylbenzenesulfonyl chloride was dissolved in 10 mL of dichloromethane, and 64 mg of 2,6-dimethylpiperidin-4-amine and 120 μL of pyridine were added and stirred at room temperature for 4 hours. Methanol was added to terminate the reaction, and dichloromethane was evaporated under reduced pressure. The resulting residue was purified by silica gel chromatography to give 158 mg of the desired compound in 80% yield. ESI (m / z): 395 (M+H) + 1 H-NMR (400 MHz, DMSO-d6) δ 7.77 (s, 1H), 7.25 (s, 2H), 4.13 (td, J = 13.3, 6.6 Hz, 2H), 3.30-3.21 (m, 2H)3.09-2.86 (m, 2H), 1.76-1.63 (bs, 1H), 1.36-1.09 (m, 28H)

[0195] (Manufacturing Example 20) 1-Ethyl-3-(trifluoromethyl)-N-(2-((2,4,6-triisopropylphenyl)sulfonamido)ethyl)-1H-pyrazole-5-carboxamide (PVZF2133) 151 mg of 2,4,6-triisopropylbenzenesulfonyl chloride was dissolved in 10 mL of dichloromethane, and 64 mg of N-(2-aminoethyl)-1-ethyl-3-trifluoromethyl-H-pyrazole-5-carboxamide and 120 μL of pyridine were added and stirred at room temperature for 4 hours. Methanol was added to terminate the reaction, and dichloromethane was evaporated under reduced pressure. The resulting residue was purified by silica gel chromatography to give 139 mg of the desired compound in 54% yield. ESI (m / z): 517 (M+H) + 1 H-NMR (400 MHz, DMSO-d6) δ 8.40 (t, J = 5.9 Hz, 1H), 7.61 (s, 1H), 7.22 (s, 2H), 7.18 (s, 1H), 4.30 (q, J = 7.2 Hz, 2H), 4.16-4.06 (m, 2H), 3.45-3.26 (m, 2H), 2.95-2.85 (m, 3H), 1.43-1.32 (m, 3H), 1.27-1.01 (m, 18H)

[0196] (Manufacturing Example 21) 2,4,6-Triisopropyl-N-(1-(5-(trifluoromethyl)pyridin-2-yl)piperidin-4-yl)benzenesulfonamide (PVZF2134) 151 mg of 2,4,6-triisopropylbenzenesulfonyl chloride was dissolved in 10 mL of dichloromethane, and 123 mg of 1-(5-(trifluoromethyl)pyridin-2-yl)piperidin-4-amine and 120 μL of pyridine were added and stirred at room temperature for 4 hours. Methanol was added to terminate the reaction, and dichloromethane was distilled off under reduced pressure. The resulting residue was purified by silica gel chromatography to give 174 mg of the target compound in a 68% yield. ESI (m / z): 512 (M+H) + 1H-NMR (400 MHz, DMSO-d6) δ 8.37 (s, 1H), 7.75 (dd, J = 9.1, 2.3 Hz, 1H), 7.65 (d, J = 7.3 Hz, 1H), 7.25 (d, J = 12.4 Hz, 2H), 6.93 (d, J = 9.0 Hz, 1H), 4.33-4.12 (m, 4H), 3.01-2.87 (m, 3H), 1.69 (d, J = 10.5 Hz, 2H), 1.39-1.30 (m, 3H), 1.22-1.03 (m, 18H)

[0197] (Manufacturing Example 22) 2,4,6-triisopropyl-N-(3-(3-(trifluoromethyl)-5,6-dihydrocyclopenta[c]pyrazol-1(4H)-yl)propyl)benzenesulfonamide (PVZF2135) 151 mg of 2,4,6-triisopropylbenzenesulfonyl chloride was dissolved in 10 mL of dichloromethane, and 117 mg of 3-(3-(trifluoromethyl)-5,6-dihydrocyclopentane[c]pyrazol-1(4H)-yl)propan-1-amine and 120 μL of pyridine were added and stirred at room temperature for 4 hours. Methanol was added to terminate the reaction, and dichloromethane was distilled off under reduced pressure. The resulting residue was purified by silica gel chromatography to obtain 155 mg of the target compound in a 62% yield. ESI (m / z): 500 (M+H) + 1 H-NMR (400 MHz, DMSO-d6) δ 7.59 (s, 1H), 7.22 (s, 2H), 4.12-3.99 (m, 4H), 2.95-2.85 (m, 1H), 2.76 (t, J = 6.2 Hz, 2H), 2.63-2.56 (m, 4H), 2.51-2.42 (m, 2H), 1.93-1.86 (m, 2H), 1.30-1.05 (m,18H)

[0198] (Manufacturing Example 23) 2,4,6-Triisopropyl-N'-(1-(3-(trifluoromethyl)benzyl)-1H-pyrazol-4-yl)benzenesulfonohydrazide (PVZF2136) 151 mg of 2,4,6-triisopropylbenzenesulfonyl chloride was dissolved in 10 mL of dichloromethane, and 128 mg of 4-hydrazinyl-1-(3-(trifluoromethyl)benzyl)-1H-pyrazole and 120 μL of pyridine were added and stirred at room temperature for 4 hours. Methanol was added to terminate the reaction, and dichloromethane was distilled off under reduced pressure. The resulting residue was purified by silica gel chromatography to give 91 mg of the desired compound in a 35% yield. ESI (m / z): 523 (M+H) + 1 H-NMR (400 MHz, DMSO-d6) δ 9.45 (s, 1H), 7.72-7.46 (m, 5H), 7.15 (s, 2H), 7.09 (s, 1H), 5.31 (s, 2H), 3.94 (t, J = 6.5 Hz, 2H), 2.93-2.82 (m, 1H), 1.23-0.87 (m, 18H)

[0199] (Manufacturing Example 24) N-(6-(2,3-difluorophenoxy)pyridin-3-yl)-2,4,6-triisopropylbenzenesulfonamide (PVZF2137) 151 mg of 2,4,6-triisopropylbenzenesulfonyl chloride was dissolved in 10 mL of dichloromethane, and 244 mg of 6-(2,3-difluorophenoxy)pyridin-3-amine and 120 μL of pyridine were added and stirred at room temperature for 4 hours. Methanol was added to terminate the reaction, and dichloromethane was evaporated under reduced pressure. The resulting residue was purified by silica gel chromatography to give 142 mg of the desired compound in 58% yield. ESI (m / z): 489 (M+H) + 1 H-NMR (400 MHz, DMSO-d6) δ 10.13 (s, 1H), 7.68 (d, J = 2.0 Hz, 1H), 7.54-7.52 (m, 1H), 7.36-7.08 (m, 6H), 3.95 (s, 2H), 2.88 (td, J = 13.7, 6.8 Hz, 1H), 1.25-0.85 (m, 18H)

[0200] (Manufacturing Example 25) 2,4,6-Triisopropyl-N-((1-((trifluoromethyl)sulfonyl)piperidin-4-yl)methyl)benzenesulfonamide (PVZF2138) 151 mg of 2,4,6-triisopropylbenzenesulfonyl chloride was dissolved in 10 mL of dichloromethane, and 123 mg of (1-((trifluoromethyl)sulfonyl)piperidin-4-yl)methanamine and 120 μL of pyridine were added and stirred at room temperature for 4 hours. Methanol was added to terminate the reaction, and dichloromethane was distilled off under reduced pressure. The resulting residue was purified by silica gel chromatography to obtain 200 mg of the target compound in a 78% yield. ESI (m / z): 513 (M+H) + 1 H-NMR (400 MHz, DMSO-d6) δ 7.65 (s, 1H), 7.24 (s, 1H), 7.23(s, 1H), 4.17-4.07 (m, 2H), 3.74 (d, J = 12.9 Hz, 2H), 3.10 (t, J = 12.4 Hz, 2H), 2.96-2.86 (m, 1H), 2.73-2.67 (m, 2H), 1.77-1.74 (m, 2H), 1.70-1.62 (m, 1H), 1.37-1.18 (m, 18H), 1.15-1.02 (m, 2H)

[0201] (Manufacturing Example 26) 2,4,6-triisopropyl-N-(1-(3-(trifluoromethyl)phenyl)-1H-pyrazol-3-yl)benzenesulfonamide (PVZF2139) 151 mg of 2,4,6-triisopropylbenzenesulfonyl chloride was dissolved in 10 mL of dichloromethane, and 114 mg of 1-(3-(trifluoromethyl)phenyl)-1H-pyrazol-3-amine and 120 μL of pyridine were added and stirred at room temperature for 4 hours. Methanol was added to terminate the reaction, and dichloromethane was distilled off under reduced pressure. The resulting residue was purified by silica gel chromatography to give 69 mg of the desired compound in a 28% yield. ESI (m / z): 494 (M+H) + 1H-NMR (400 MHz, DMSO-d6) δ 10.85 (s, 1H), 8.49 (s, 1H), 7.95-7.92 (m, 2H), 7.66 (t, J = 7.9 Hz, 1H), 7.57 (d, J = 7.6 Hz, 1H), 7.20 (s, 2H), 6.09 (d, J = 2.2 Hz, 1H), 4.27 (s, 2H), 2.93-2.83 (m, 1H), 1.16 (dd, J = 6.7, 2.3 Hz, 18H)

[0202] (Manufacturing Example 27) (E)-N'-Hydroxy-3-(3-methyl-5-(trifluoromethyl)-1H-pyrazol-1-yl)-N-((2,4,6-triisopropylphenyl)sulfonyl)propanimidamide (PVZF2140) 151 mg of 2,4,6-triisopropylbenzenesulfonyl chloride was dissolved in 10 mL of dichloromethane, and 118 mg of (E)-2-amino-4-(3-methyl-5-(trifluoromethyl)-1H-pyrazol-1-yl)but-1-en-1-ol and 120 μL of pyridine were added and stirred at room temperature for 4 hours. Methanol was added to terminate the reaction, and dichloromethane was distilled off under reduced pressure. The resulting residue was purified by silica gel chromatography to obtain 53 mg of the desired compound in a 21% yield. ESI (m / z): 503 (M+H) + 1 H-NMR (400 MHz, DMSO-d6) δ 8.55 (s, 1H), 7.23 (s, 2H), 6.82 (s, 2H), 6.55 (s, 1H), 4.15-4.06 (m, 4H), 3.57 (d, J = 1.2 Hz, 1H), 2.97-2.87 (m, 1H), 2.11 (s, 3H), 1.21-1.15 (m, 18H)

[0203] (Manufacturing Example 28) N-(3-(3-cyclopropyl-4,5-dihydroxy-2-oxo-2,3-dihydro-1H-imidazol-1-yl)phenyl)-2,4,6-triisopropylbenzenesulfonamide (PVZF2141) 151 mg of 2,4,6-triisopropylbenzenesulfonyl chloride was dissolved in 10 mL of dichloromethane, and 137 mg of 1-(3-aminophenyl)-3-cyclopropyl-4,5-dihydroxy-1,3-dihydro-2H-imidazol-2-one and 120 μL of pyridine were added and stirred at room temperature for 4 hours. Methanol was added to terminate the reaction, and the dichloromethane was evaporated under reduced pressure. The resulting residue was purified by silica gel chromatography to give 108 mg of the desired compound in 42% yield. ESI (m / z): 514 (M+H) + 1 H-NMR (400 MHz, DMSO-d6) δ 10.29 (s, 1H), 7.49 (s, 1H), 7.28-7.07 (m, 5H), 6.79 (d, J = 8.0 Hz, 1H), 5.44 (d, J = 9.8 Hz, 1H), 4.20 (s, 2H), 3.57 (d, J = 1.2 Hz, 1H), 2.89 (td, J = 13.6, 6.6 Hz, 1H), 1.18-1.13 (m, 18H), 0.90-0.79 (m, 4H)

[0204] (Manufacturing Example 29) 2,4,6-triisopropyl-N-(((2,2,6,6-tetramethylpiperidin-4-yl)methyl)benzenesulfonamide (PVZF2142) 151 mg of 2,4,6-triisopropylbenzenesulfonyl chloride was dissolved in 10 mL of dichloromethane, and 135 mg of (2,2,6,6-tetramethylpiperidin-4-yl)methanamine and 120 μL of pyridine were added and stirred at room temperature for 4 hours. Methanol was added to terminate the reaction, and dichloromethane was evaporated under reduced pressure. The resulting residue was purified by silica gel chromatography to give 194 mg of the desired compound in 89% yield. ESI (m / z): 437 (M+H) + 1H-NMR (400 MHz, DMSO-d6) 1H-NMR (400 MHz, DMSO) δ 7.80-7.67 (m, 1H), 7.37-7.18 (m, 2H), 6.95 (s, 1H), 4.17-4.07 (m, 2H), 2.96-2.86 (m, 1H), 2.75 (t, J = 6.2 Hz, 2H), 1.91-1.86 (m, 1H), 1.61-1.43 (m, 2H), 1.26-1.15 (m, 32H)

[0205] (Manufacturing Example 30) 2,4,6-triisopropyl-N-(1,2,3,4-tetrahydroquinolin-4-yl)benzenesulfonamide (PVZF2143) 151 mg of 2,4,6-triisopropylbenzenesulfonyl chloride was dissolved in 10 mL of dichloromethane, and 124 mg of 1,2,3,4-tetrahydroquinolin-4-amine and 120 μL of pyridine were added and stirred at room temperature for 4 hours. Methanol was added to terminate the reaction, and the dichloromethane was evaporated under reduced pressure. The resulting residue was purified by silica gel chromatography to give 155 mg of the desired compound in 75% yield. ESI (m / z): 415 (M+H) + 1 H-NMR (400 MHz, DMSO-d6) δ 8.00 (d, J = 8.8 Hz, 1H), 7.24 (s, 2H), 6.94-6.86 (m, 1H), 6.70 (d, J = 7.6 Hz, 1H), 6.47-6.32 (m, 2H), 5.84 (s, 1H), 4.34-4.30 (m, 1H), 4.22-4.10 (m, 2H), 3.22-3.17 (m, 1H), 3.06 (d, J = 12.0 Hz, 1H), 2.97-2.90 (m, 1H), 1.80-1.65 (m, 2H), 1.32-1.16 (m, 18H)

Claims

1. An RNA methyltransferase inhibitor comprising a pyrazoline compound represented by the following general formula (2): 【Chemistry 1】 [In formula (2), n represents an integer of 2 to 4. 4 are the same or different and represent any one of the following groups (4-1) to (4-34). (4-1) phenyl group, (4-2) a phenylsulfonyl group, (4-3) alkylcarbonyl group, (4-4) aminothiocarbonyl group, (4-5) benzodioxolyl group, (4-6) an alkylsulfonyl group, (4-7) adamantylcarbonyl group, (4-8) benzopyrazyl group, (4-9) phenylcarbonyl group, (4-10) naphthyl group, (4-11) Furylcarbonyl group, (4-12) thienylcarbonyl group, (4-13) quinazolyl group, (4-14) quinoxalyl group, (4-15) Hydroxyl group, (4-16) an alkenyl group, (4-17) thiazolyl group, (4-18) cycloalkylcarbonyl group, (4-19) aminocarbonyl group, (4-20) furyl group, (4-21) thienyl group, (4-22) pyridyl group, (4-23) a cycloalkenyl group, (4-24) alkyl group, (4-25) a pyrazolyl group, (4-26) quinolyl group, (4-27) alkenylcarbonyl group, (4-28) Benzopyranyl group, (4-29) a benzopyrimidyl group, (4-30) pyrrolidinoalkylcarbonyl group, (4-31) quinolylcarbonyl group, (4-32) an alkoxycarbonyl group, (4-33) morpholino group, (4-34) a pyrrolidinocarbonylalkoxy group, (4-35) benzodioxy-6-yl group; Each of the groups represented by (4-1) to (4-35) may further have a substituent. The bond between the 4-position carbon atom and the 5-position carbon atom of the pyrazole skeleton represents a single bond or a double bond. Alternatively, two adjacent carbon atoms constituting the pyrazoline ring may be bonded to each other to form a ring. Alternatively, the nitrogen atom constituting the pyrazoline ring and the carbon atom adjacent thereto may be bonded to each other to form a ring.

2. The RNA methyltransferase inhibitor according to claim 1, wherein the substituent on the phenyl group represented by (4-1) is at least one selected from the group consisting of halogen, an alkyl group, a haloalkyl group, an alkoxy group, a hydroxyl group, an alkylsulfonylamino group, a nitro group, an amino group, a carboxyl group, and a phenyl group.

3. The substituent on the alkylcarbonyl group represented by (4-3) is a phenylalkylamino group, a triazolylthio group, a phenoxy group, an oxadiazolylthio group, an ester group, a piperazinyl group, a carboxyl group, a pyrimidinylthio group, a quinazolyloxy group, a morpholinocarbonyl group, a morpholino group, a benzotriazolyl group, a pyrazolylcarbonyl group, a pyrimidyl group, a pyrrolidino group, a piperidino group, a tetrahydroimidazolyl group, a halogen atom, a naphthyloxy group, an alkoxy group, an imidazolyl group, a tetrazolylthio group, an alkylamino group, a pyridyl group, a tetrazolyl group, a benzodioxonyloxy group, an aminocarbonyl group, or a piperazinyl group.

3. The RNA methyltransferase inhibitor according to claim 1, which is at least one selected from the group consisting of a phenylalkylthio group, an alkylcarbonyloxy group, a benzotriazolylthio group, a pyridazinyl group, a pyrrolylcarbonyloxy group, a piperidino group, a dihydrothiazolylthio group, a benzopyrazyl group, a thienopyridinoxy group, a thienopyrimidinylthio group, a cyclopentathienopyrimidinyl group, a thiadiazolylthio group, an azepinylthio group, a dioxoloquinolinyl group, a diazaspirononanyl group, an imidazolidinyl group, a triazolylthio group, a dihydropyridazinyl group and a 1,3-diazaspirondecanyl group.

4. The RNA methyltransferase inhibitor according to any one of claims 1 to 3, which is used in the treatment of cancer.

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