5-Substituted pyridin-2(1H)-one compounds and their uses

Abstract

The present invention relates to a series of 5-substituted pyridin-2(1H)-one compounds and uses thereof, in particular to a compound represented by formula (X) or a pharma- ceutically acceptable salt thereof: JPEG2024540069000060.jpg4466

Description

[Technical Field]

[0001] This application claims priority to: CN202111272187.7, October 29, 2021, CN202210495439.0, May 7, 2022.

[0002] The present invention relates to a series of 5-substituted pyridin-2(1H)-one compounds and uses thereof, particularly to a compound represented by formula (X) or a pharmaceutically acceptable salt thereof: [Background technology]

[0003] Porcupine (PORCN) protein is an acyltransferase that activates the extracellular secretion of Wnt proteins through palmitoylation of WNT proteins. Extracellular Wnt proteins bind to Frizzled receptors on the cell membrane, activating the Wnt / β-catenin pathway, which causes β-catenin to accumulate and enter the nucleus, where it binds to TCF / LEF transcription factors and regulates the transcription of downstream target genes, promoting the proliferation and activation of tumor cells.

[0004] The Wnt / β-catenin signaling pathway is overactivated in many tumors, including gastric, colorectal, liver, and pancreatic cancers. Porcupine inhibitors can inhibit porcupine protein activity and block palmitoylation and extracellular secretion of Wnt proteins, thereby inhibiting the abnormal activation of the Wnt / β-catenin signaling pathway and inhibiting the proliferation of various tumor cells. Porcupine inhibitors currently in clinical trials include LGK974 (patent application for this compound: WO2010101849) and ETC-1922159 (patent application for this compound: WO2014189466). These porcupine inhibitors in clinical development have issues such as rapid metabolism, short half-life, and significant side effects. Therefore, the development of novel porcupine inhibitors that regulate Wnt / β-catenin pathway signaling is of great clinical and social value. [ka] Summary of the Invention

[0005] In one aspect, the present invention provides a compound represented by formula (X) or a pharmaceutically acceptable salt thereof: [ka] however, T is selected from CR or N; T1 is selected from CH or N; T2 is selected from CH or N; T3 is selected from CH or N; L is selected from a single bond and —C(R4R5)—; Ring A is a 9-10 membered heteroaryl, wherein said 9-10 membered heteroaryl optionally has 1, 2, or 3 R a is replaced by Each R1 is independently H, F, Cl, Br, I, -CN, -OH, -NH2, C 1-3 Alkyl, C 1-3Alkoxy and C 1-3 alkylamino, wherein said C 1-3 Alkyl, C 1-3 Alkoxy and C 1-3 The alkylamino may each independently optionally contain 1, 2, or 3 R b is replaced by Each R2 is independently H, F, Cl, Br, I, -CN, -OH, -NH2, C 1-3 Alkyl, C 1-3 Alkoxy and C 1-3 alkylamino, wherein said C 1-3 Alkyl, C 1-3 Alkoxy and C 1-3 The alkylamino may each independently optionally contain 1, 2, or 3 R c is replaced by Each R3 is independently H, F, Cl, Br, I, -CN, -OH, -NH2, C 1-3 Alkyl, C 1-3 Alkoxy and C 1-3 alkylamino, wherein said C 1-3 Alkyl, C 1-3 Alkoxy and C 1-3 The alkylamino may each independently optionally contain 1, 2, or 3 R d is replaced by R4 and R5 are each independently H and C 1-3 alkyl, wherein said C 1-3 The alkyl may optionally contain one, two, or three R e is replaced by R is H, F, Cl, Br, I, -CN, -OH, -NH2, C 1-3 Alkyl, C 1-3 Alkoxy and C 1-3 alkylamino, wherein said C 1-3 Alkyl, C 1-3 Alkoxy and C 1-3 The alkylamino may each independently optionally contain 1, 2, or 3 R f is replaced by R aare each independently selected from F, Cl, Br, I, —OH, —NH and —CN; R b are each independently selected from F, Cl, Br, I, —OH, —NH and —CN; R c are each independently selected from F, Cl, Br, I, —OH, —NH and —CN; R d are each independently selected from F, Cl, Br, I, —OH, —NH and —CN; R e are each independently selected from F, Cl, Br, I, —OH, —NH and —CN; R f are each independently selected from F, Cl, Br, I, —OH, —NH and —CN; m is selected from 0, 1, 2 and 3; n is selected from 0, 1, 2 and 3; p is selected from 0, 1, 2 and 3; The "hetero" in the 9- to 10-membered heteroaryl represents 1, 2, 3, or 4 heteroatoms or heteroatom groups independently selected from --O--, --S--, and --N--.

[0006] In one aspect, the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof. [ka] however, T is selected from CR or N; T1 is selected from CH or N; T2 is selected from CH or N; L is selected from a single bond and —C(R4R5)—; Ring A is a 9-10 membered heteroaryl, wherein said 9-10 membered heteroaryl optionally has 1, 2, or 3 R a is replaced by Each R1 is independently H, F, Cl, Br, I, -CN, -OH, -NH2, C 1-3 Alkyl, C1-3 Alkoxy, and C 1-3 alkylamino, wherein said C 1-3 Alkyl, C 1-3 Alkoxy and C 1-3 The alkylamino may each independently optionally contain 1, 2, or 3 R b is replaced by Each R2 is independently H, F, Cl, Br, I, -CN, -OH, -NH2, C 1-3 Alkyl, C 1-3 Alkoxy and C 1-3 alkylamino, wherein said C 1-3 Alkyl, C 1-3 Alkoxy and C 1-3 The alkylamino may each independently optionally contain 1, 2, or 3 R c is replaced by Each R3 is independently H, F, Cl, Br, I, -CN, -OH, -NH2, C 1-3 Alkyl, C 1-3 Alkoxy and C 1-3 alkylamino, wherein said C 1-3 Alkyl, C 1-3 Alkoxy and C 1-3 The alkylamino may each independently optionally contain 1, 2, or 3 R d is replaced by R4 and R5 are each independently H and C 1-3 alkyl, wherein said C 1-3 The alkyl may optionally contain one, two, or three R e is replaced by R is H, F, Cl, Br, I, -CN, -OH, -NH2, C 1-3 Alkyl, C 1-3 Alkoxy and C 1-3 alkylamino, wherein said C 1-3 Alkyl, C 1-3 Alkoxy and C 1-3 The alkylamino may each independently optionally contain 1, 2, or 3 R f is replaced by R aare each independently selected from F, Cl, Br, I, —OH, —NH and —CN; R b are each independently selected from F, Cl, Br, I, —OH, —NH and —CN; R c are each independently selected from F, Cl, Br, I, —OH, —NH and —CN; R d are each independently selected from F, Cl, Br, I, —OH, —NH and —CN; R e are each independently selected from F, Cl, Br, I, —OH, —NH and —CN; R f are each independently selected from F, Cl, Br, I, —OH, —NH and —CN; m is selected from 0, 1, 2 and 3; n is selected from 0, 1, 2 and 3; p is selected from 0, 1, 2 and 3; The "hetero" in the 9- to 10-membered heteroaryl represents 1, 2, 3, or 4 heteroatoms or heteroatom groups independently selected from --O--, --S--, and --N--.

[0007] In some embodiments of the present invention, the compound has a structure represented by formula (X-1). [ka] wherein ring A, R1, R2, R3, R4, R5, T, T1, T2, T3, m, n, and p are as defined in the present invention.

[0008] In some embodiments of the present invention, the compound has a structure represented by formula (I-1). [ka] wherein ring A, R1, R2, R3, R4, R5, T, T1, T2, m, n, and p are as defined in the present invention.

[0009] In some embodiments of the invention, each R is independently selected from H, F, Cl, Br, I, —CN, —OH, —NH, —CH, and —OCH, wherein —CH and —OCH are each independently optionally joined to one, two, or three R b is replaced by R b and other variables as defined in the present invention.

[0010] In some embodiments of the present invention, T is selected from CR, where R and other variables are as defined herein.

[0011] In some embodiments of the present invention, T above is selected from N, and other variables are as defined herein.

[0012] In some embodiments of the present invention, T1 above is selected from CH, and other variables are as defined herein.

[0013] In some embodiments of the present invention, T1 above is selected from N, and other variables are as defined herein.

[0014] In some embodiments of the present invention, T2 above is selected from CH, and other variables are as defined herein.

[0015] In some embodiments of the present invention, T2 above is selected from N, and other variables are as defined herein.

[0016] In some embodiments of the present invention, T3 above is selected from CH, and other variables are as defined herein.

[0017] In some embodiments of the present invention, T3 above is selected from N, and other variables are as defined herein.

[0018] In some embodiments of the invention, each R is independently selected from H, F, Cl, Br, I, —CN, —OH, —NH, and —CH, wherein said —CH is optionally selected from 1, 2, or 3 R b is replaced by R b and other variables as defined in the present invention.

[0019] In some embodiments of the present invention, each R 1 above is independently selected from H, F, —CH 3 and —OCH 3 , and all other variables are as defined herein.

[0020] In some embodiments of the present invention, each R 1 above is independently selected from H and F, and all other variables are as defined herein.

[0021] In some embodiments of the present invention, each R1 above is independently selected from F, and all other variables are as defined herein. In some embodiments of the present invention, each R2 above is independently selected from H, and all other variables are as defined herein.

[0022] In some embodiments of the invention, each R is independently selected from H, F, Cl, Br, I, —CN, —OH, —NH, and —CH, wherein said —CH is optionally selected from 1, 2, or 3 R d is replaced by R d and other variables as defined in the present invention.

[0023] In some embodiments of the present invention, each R3 above is independently selected from -CH3, and all other variables are as defined herein.

[0024] In some embodiments of the present invention, R and R are each independently selected from H and —CH, wherein said —CH is optionally selected from 1, 2, or 3 R e is replaced by R eand other variables as defined in the present invention.

[0025] In some embodiments of the present invention, R4 and R5 above are each independently selected from H, and all other variables are as defined herein.

[0026] In some embodiments of the invention, R is selected from H, F, Cl, Br, I, —CN, —OH, —NH and —CH, wherein said —CH is optionally joined by 1, 2 or 3 R f is replaced by R f and other variables as defined in the present invention.

[0027] In some embodiments of the present invention, R above is selected from H and F, and other variables are as defined herein.

[0028] In some embodiments of the present invention, R is selected from F, and other variables are as defined herein.

[0029] In some embodiments of the present invention, m above is selected from 0, and the other variables are as defined herein.

[0030] In some embodiments of the present invention, m above is selected from 1, and the other variables are as defined herein.

[0031] In some embodiments of the present invention, m above is selected from 2, and the other variables are as defined herein.

[0032] In some embodiments of the present invention, ring A above is [ka] is selected from, where: [ka] each independently optionally 1, 2 or 3 Ra is replaced by R a and other variables as defined in the present invention.

[0033] In some embodiments of the present invention, ring A above is [ka] and the other variables are as defined in the present invention.

[0034] In some embodiments of the present invention, the compound has a structure represented by formula (X-2) or (X-3).

[0035] [ka] however, E is selected from CH and N; E1 is selected from CH and N; R1, R2, R3, R4, R5, T, T1, T2, T3, m, n and p are as defined in the present invention.

[0036] In some embodiments of the present invention, the compound has a structure represented by formula (I-2) or (I-3).

[0037] [ka] however, E is selected from CH and N; E1 is selected from CH and N; R1, R2, R3, R4, R5, T, T1, T2, m, n and p are as defined in the present invention.

[0038] In some embodiments of the present invention, the compound has a structure represented by formula (X-4), (X-5), or (X-6).

[0039] [ka] wherein R1, R3, T, T1, T2, T3, m and p are as defined in the present invention.

[0040] In some embodiments of the present invention, the compound has a structure represented by formula (I-4), (I-5), or (I-6).

[0041] [ka] wherein R1, R3, T, T1, T2, m and p are as defined in the present invention.

[0042] In some embodiments of the present invention, the compound has the formula (X-7), (X-8) or It has a structure represented by (X-9).

[0043] [ka] wherein R1, R3, T, T1, T2 and T3 are as defined in the present invention.

[0044] In some embodiments of the present invention, the compound has formula (I-7), (I-8) or It has the structure represented by (I-9).

[0045] [ka] wherein R1, R3, T, T1 and T2 are as defined in the present invention.

[0046] In some embodiments of the present invention, the compound has a structure represented by formula (X-4A), (X-4B), (X-4C), (X-4D), (X-5A), or (X-6A).

[0047] [ka] wherein R1, R3, T, m and p are as defined in the present invention.

[0048] In some embodiments of the present invention, the compound has a structure represented by formula (X-4A-1), (X-4B-1), (X-4C-1), (X-4D-1), (X-5A-1), or (X-6A-1).

[0049] [ka] wherein R1, R3, T and m are as defined in the present invention.

[0050] Further embodiments of the present invention are formed by any combination of the above variables.

[0051] The present invention further provides a compound of the formula: or a pharmaceutically acceptable salt thereof:

[0052] [ka] The present invention further provides use of the above compound or a pharmaceutically acceptable salt thereof in the preparation of a porcupine inhibitor.

[0053] The present invention further provides the use of the above compound or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating pancreatic cancer, colorectal cancer and gastric cancer.

[0054] The present invention provides a type A crystal of Compound 2, characterized in that the powder X-ray diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 7.45±0.20°, 13.53±0.20°, 13.94±0.20°, and 15.93±0.20°. [ka] In some embodiments of the present invention, the above-mentioned Type A crystals are characterized in that their powder X-ray diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 7.45±0.20°, 10.56±0.20°, 13.53±0.20°, 13.94±0.20°, 14.86±0.20°, 15.93±0.20°, and 17.96±0.20°.

[0055] In some embodiments of the present invention, the above-mentioned Type A crystal is characterized by having at least 4, 5, 6, or 7 characteristic diffraction peaks selected from 7.45±0.20°, 10.56±0.20°, 13.53±0.20°, 13.94±0.20°, 14.86±0.20°, 15.93±0.20°, and 17.96±0.20° in a powder X-ray diffraction pattern expressed in 2θ angles.

[0056] In some embodiments of the present invention, the above-mentioned type A crystal has a powder X-ray diffraction pattern having characteristic diffraction peaks at the following 2θ angles: 4.96±0.20°, 7.45±0.20°, 10.56±0.20°, 13.53±0.20°, 13.94±0.20°, 14.86±0.20°, 15.93±0.20°, 17.96±0.20°, 19.87±0.20°, 20.90±0.20°, 25.38±0.20°, and 28.02±0.20°.

[0057] In some embodiments of the present invention, the above-mentioned Type A crystal is characterized by having, in a powder X-ray diffraction pattern expressed in 2θ angles, at least 4, 5, 6, 7, 8, 9, 10, 11, or 12 characteristic diffraction peaks selected from 4.96±0.20°, 7.45±0.20°, 10.56±0.20°, 13.53±0.20°, 13.94±0.20°, 14.86±0.20°, 15.93±0.20°, 17.96±0.20°, 19.87±0.20°, 20.90±0.20°, 25.38±0.20°, and 28.02±0.20°.

[0058] In some embodiments of the present invention, the above-mentioned type A crystal has a powder X-ray diffraction pattern having characteristic diffraction peaks at the following 2θ angles: 4.96±0.20°, 7.45±0.20°, 10.56±0.20°, 13.53±0.20°, 13.94±0.20°, 14.86±0.20°, 15.93±0.20°, 17.96±0.20°, 19.06±0.20°, 19.87±0.20°, 20.90±0.20°, 24.11±0.20°, 24.97±0.20°, 25.38±0.20°, and 28.02±0.20°.

[0059] In some embodiments of the present invention, the above-mentioned Form A crystal has a powder X-ray diffraction pattern exhibiting the following 2θ angles: 4.96±0.20°, 7.45±0.20°, 10.56±0.20°, 13.53±0.20°, 13.94±0.20°, 14.86±0.20°, 15.93±0.20°, 17.56±0.20°, 17.96±0.20°, 19.06±0.20°, 19.87±0.20°, 20.90±0.20°, 21.21±0.20°, and 21.90±0.20°. , 22.63±0.20°, 24.11±0.20°, 24.97±0.20°, 25.38±0.20°, 25.96±0.20°, 27.14±0.20°, 28.02±0.20°, 28.73±0.20°, 29.94±0.20°, 30.89±0.20°, 32.50±0.20°, 34.01±0.20°, 35.05±0.20°, 35.95±0.20°, 37.54±0.20° and 39.08±0.20°.

[0060] In some embodiments of the present invention, the above-mentioned Form A crystal has a powder X-ray diffraction pattern having characteristic diffraction peaks at the following 2θ angles: 4.96°, 7.45°, 10.56°, 13.53°, 13.94°, 14.86°, 15.93°, 17.56°, 17.96°, 19.06°, 19.87°, 20.90°, 21.21°, 21.90°, 22.63°, 24.11°, 24.97°, 25.38°, 25.96°, 27.14°, 28.02°, 28.73°, 29.94°, 30.89°, 32.50°, 34.01°, 35.05°, 35.95°, 37.54°, and 39.08°.

[0061] In some embodiments of the present invention, the above-mentioned Form A crystal has a powder X-ray diffraction pattern exhibiting the following 2θ angles: 13.53±0.20°, 13.94±0.20°, 15.93±0.20°, and / or 4.96±0.20°, and / or 7.45±0.20°, and / or 10.56±0.20°, and / or 14.86±0.20°, and / or 17.56±0.20°, and / or 17.96±0.20°, and / or 19.06±0.20°, and / or 19.87±0.20°, and / or 20.90±0.20°, and / or 21.21±0.20°, and / or 21.90±0.20°, and / or 22.63±0.20°. and / or 24.11±0.20°, and / or 24.97±0.20°, and / or 25.38±0.20°, and / or 25.96±0.20°, and / or 27.14±0.20°, and / or 28.02±0.20°, and / or 28.73±0.20°, and / or 29.94±0.20°, and / or 30.89±0.20°, and / or 32.50±0.20°, and / or 34.01±0.20°, and / or 35.05±0.20°, and / or 35.95±0.20°, and / or 37.54±0.20°, and / or 39.08±0.20°.

[0062] In some embodiments of the present invention, the XRPD pattern of the above-mentioned Form A crystals is essentially as shown in FIG.

[0063] In some embodiments of the present invention, the analytical data of the XRPD pattern of the above-mentioned Form A crystal is as shown in Table 1.

[0064] [Table 1] In some embodiments of the present invention, the differential scanning calorimetry curve for the above-mentioned Type A crystals has endothermic peak onsets at 259.7°C±5°C and 274.7°C±5°C.

[0065] In some embodiments of the present invention, the DSC pattern of the above-mentioned Form A crystals is essentially as shown in FIG.

[0066] In some embodiments of the present invention, the thermogravimetric analysis curve of the above-mentioned A-type crystals shows a weight loss of 0.80% at 240°C ± 3°C.

[0067] In some embodiments of the present invention, the TGA pattern of the above-mentioned Form A crystals is essentially as shown in FIG.

[0068] In some embodiments of the present invention, the DVS isotherm pattern of the above-mentioned Form A crystals is essentially as shown in FIG.

[0069] The present invention further provides a crystalline form B of compound 2 of formula, characterized in that the powder X-ray diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 13.43±0.20°, 14.07±0.20°, and 15.07±0.20°. [ka] In some embodiments of the present invention, the type B crystals have a powder X-ray diffraction pattern having characteristic diffraction peaks at the following 2θ angles: 11.72±0.20°, 13.43±0.20°, 14.07±0.20°, 15.07±0.20°, 16.39±0.20°, 18.21±0.20°, 22.53±0.20°, and 25.23±0.20°.

[0070] In some embodiments of the present invention, the above-mentioned type B crystal is characterized by having at least 4, 5, 6, 7, or 8 characteristic diffraction peaks selected from 11.72±0.20°, 13.43±0.20°, 14.07±0.20°, 15.07±0.20°, 16.39±0.20°, 18.21±0.20°, 22.53±0.20°, and 25.23±0.20° in a powder X-ray diffraction pattern expressed in 2θ angles.

[0071] In some embodiments of the present invention, the type B crystals have a powder X-ray diffraction pattern having characteristic diffraction peaks at the following 2θ angles: 4.86±0.20°, 7.05±0.20°, 11.72±0.20°, 13.43±0.20°, 14.07±0.20°, 14.54±0.20°, 15.07±0.20°, 16.39±0.20°, 18.21±0.20°, 19.67±0.20°, 22.53±0.20°, and 25.23±0.20°.

[0072] In some embodiments of the present invention, the type B crystal is characterized by having, in a powder X-ray diffraction pattern expressed in 2θ angles, at least 4, 5, 6, 7, 8, 9, 10, 11, or 12 characteristic diffraction peaks selected from 4.86±0.20°, 7.05±0.20°, 11.72±0.20°, 13.43±0.20°, 14.07±0.20°, 14.54±0.20°, 15.07±0.20°, 16.39±0.20°, 18.21±0.20°, 19.67±0.20°, 22.53±0.20°, and 25.23±0.20°.

[0073] In some embodiments of the present invention, the type B crystals have a powder X-ray diffraction pattern having characteristic diffraction peaks at the following 2θ angles: 4.86±0.20°, 7.05±0.20°, 11.72±0.20°, 13.43±0.20°, 14.07±0.20°, 14.50±0.20°, 15.07±0.20°, 16.39±0.20°, 18.21±0.20°, 19.67±0.20°, 22.53±0.20°, 23.24±0.20°, 24.28±0.20°, 25.23±0.20°, 28.32±0.20°, and 29.39±0.20°.

[0074] In some embodiments of the present invention, the type B crystals have a powder X-ray diffraction pattern exhibiting the following 2θ angles: 4.86±0.20°, 7.05±0.20°, 11.72±0.20°, 13.43±0.20°, 14.07±0.20°, 14.50±0.20°, 15.07±0.20°, 15.97±0.20°, 16.39±0.20°, 18.21±0.20°, 19.39±0.20°, 19.67±0.20°, 21.18±0.20°, 22.53±0.20°, 23.24±0.20°. ±0.20°, 24.28±0.20°, 25.23±0.20°, 25.79±0.20°, 26.46±0.20°, 27.20±0.20°, 27.90±0.20°, 28.32±0.20°, 28.90±0.20°, 29.39±0.20°, 31.70±0.20°, 32.54±0.20°, 33.50±0.20°, 34.34±0.20°, 34.90±0.20°, 37.53±0.20° and 38.96±0.20°.

[0075] In some embodiments of the present invention, the type B crystals have a powder X-ray diffraction pattern exhibiting the following 2θ angles: 4.86±0.10°, 7.05±0.10°, 11.72±0.10°, 13.43±0.10°, 14.07±0.10°, 14.50±0.10°, 15.07±0.10°, 15.97±0.10°, 16.39±0.10°, 18.21±0.10°, 19.39±0.10°, 19.67±0.10°, 21.18±0.10°, 22.53±0.10°, 23.24±0.10°, 24.86±0.10°, 25.05±0.10°, 26.05±0.10°, 27.05±0.10°, 28.05±0.10°, 29.05±0.10°, 30.05±0.10°, 31.05±0.10°, 32.05±0.10°, 33.05±0.10°, 34.05±0.10°, 35.05±0.10°, 36.05±0.10°, 37.05±0.10°, 38.05±0.10°, 39.05±0.10°, 40.05±0.10°, 41.05±0.10°, 42.05±0.10°, 43.05±0.10°, 44.05±0.10°, 45.05±0.10°, 4 ±0.10°, 24.28±0.10°, 25.23±0.10°, 25.79±0.10°, 26.46±0.10°, 27.20±0.10°, 27.90±0.10°, 28.32±0.10°, 28.90±0.10°, 29.39±0.10°, 31.70±0.10°, 32.54±0.10°, 33.50±0.10°, 34.34±0.10°, 34.90±0.10°, 37.53±0.10° and 38.96±0.10°.

[0076] In some embodiments of the present invention, the type B crystals have a powder X-ray diffraction pattern having characteristic diffraction peaks at the following 2θ angles: 4.86°, 7.05°, 11.72°, 13.43°, 14.07°, 14.50°, 15.07°, 15.97°, 16.39°, 18.21°, 19.39°, 19.67°, 21.18°, 22.53°, 23.24°, 24.28°, 25.23°, 25.79°, 26.46°, 27.20°, 27.90°, 28.32°, 28.90°, 29.39°, 31.70°, 32.54°, 33.50°, 34.34°, 34.90°, 37.53°, and 38.96°.

[0077] In some embodiments of the present invention, the type B crystals have a powder X-ray diffraction pattern at the following 2θ angles: 13.43±0.20°, 14.07±0.20°, 15.07±0.20°, and / or 4.86±0.20°, and / or 7.05±0.20°, and / or 11.72±0.20°, and / or 14.50±0.20°, and / or 15.97±0.20°, and / or 16.39±0.20°, and / or 18.21±0.20°, and / or 19.39±0.20°, and / or 19.67±0.20°, and / or 21.18±0.20°, and / or 22.53±0.20°, and / or 23.24±0.20°, and / or or 24.28±0.20°, and / or 25.23±0.20°, and / or 25.79±0.20°, and / or 26.46±0.20°, and / or 27.20±0.20°, and / or 27.90±0.20°, and / or 28.32±0.20°, and / or 28.90±0.20°, and / or 29.39±0.20°, and / or 31.70±0.20°, and / or 32.54±0.20°, and / or 33.50±0.20°, and / or 34.34±0.20°, and / or 34.90±0.20°, and / or 37.53±0.20°, and / or 38.96±0.20°.

[0078] In some embodiments of the present invention, the type B crystals have a powder X-ray diffraction pattern at the following 2θ angles: 13.43±0.10°, 14.07±0.10°, 15.07±0.10°, and / or 4.86±0.10°, and / or 7.05±0.10°, and / or 11.72±0.10°, and / or 14.50±0.10°, and / or 15.97±0.10°, and / or 16.39±0.10°, and / or 18.21±0.10°, and / or 19.39±0.10°, and / or 19.67±0.10°, and / or 21.18±0.10°, and / or 22.53±0.10°, and / or 23.24±0.10°, and / or or 24.28±0.10°, and / or 25.23±0.10°, and / or 25.79±0.10°, and / or 26.46±0.10°, and / or 27.20±0.10°, and / or 27.90±0.10°, and / or 28.32±0.10°, and / or 28.90±0.10°, and / or 29.39±0.10°, and / or 31.70±0.10°, and / or 32.54±0.10°, and / or 33.50±0.10°, and / or 34.34±0.10°, and / or 34.90±0.10°, and / or 37.53±0.10°, and / or 38.96±0.10°.

[0079] In some embodiments of the present invention, the XRPD pattern of the above-mentioned Form B crystals is essentially as shown in FIG.

[0080] In some embodiments of the present invention, the analytical data of the XRPD pattern of the above-mentioned B-type crystals are as shown in Table 2.

[0081] [Table 2] In some embodiments of the present invention, the differential scanning calorimetry curve of the above-mentioned B-type crystals has an endothermic peak onset at 275.1°C ± 5°C.

[0082] In some embodiments of the present invention, the DSC pattern of the above-mentioned B-type crystals is essentially as shown in FIG.

[0083] In some embodiments of the present invention, the thermogravimetric analysis curve of the B-type crystals shows a weight loss of 0.77% at 260°C ± 3°C.

[0084] In some embodiments of the present invention, the TGA pattern of the above-mentioned B-type crystals is essentially as shown in FIG.

[0085] The present invention further provides use of the above-mentioned type A crystals and / or type B crystals in the preparation of a porcupine inhibitor.

[0086] The present invention further provides use of the above-mentioned Form A crystals and / or Form B crystals in the preparation of a medicament for treating pancreatic cancer, colorectal cancer, and gastric cancer.

[0087] The present invention further provides use of the above compound or a pharmaceutically acceptable salt thereof, or / and type A crystals of Compound 2, or / and type B crystals of Compound 2, in the preparation of a porcupine inhibitor.

[0088] The present invention further provides use of the above compound or a pharmaceutically acceptable salt thereof, or / and type A crystals of Compound 2, or / and type B crystals of Compound 2, in the preparation of a medicament for treating pancreatic cancer, colorectal cancer, and gastric cancer.

[0089] The present invention further provides methods for biological testing of the above compounds.

[0090] Experimental test method 1: In vivo efficacy test in Capan-2 nude rat tumor xenografts Capan-2 cells were cultured in McCoy's 5A medium containing 10% fetal bovine serum (FBS) in a 5% CO2, 37°C incubator. Subcultured to an appropriate density, tumor cells were harvested, counted, and resuspended in McCoy's 5A medium at a cell suspension concentration of 2 × 10 7 / mL and used for inoculation.

[0091] Establishment of human pancreatic cancer Capan-2 xenograft tumors: Cells were harvested and grown at a concentration of 2 x 10 7 The tumor cells were adjusted to 2 × 10 cells / mL (resuspended in McCoy's 5A medium containing 50% Martigel) and injected subcutaneously into the right dorsum of mice under sterile conditions, with the number of cells inoculated per mouse being 2 × 10 cells. 6 After the tumor has grown to a predetermined size, the length (a) and width (b) of the tumor are measured using digital calipers, and the tumor volume is calculated using the formula TV = a × b 2 / 2.

[0092] Capan-2 tumor cells were inoculated, and tumor size was approximately 150 mm 3 When the animals reached maturity, they were divided into groups of 6 animals each. The day of grouping was designated as day 0, and administration began on the day of grouping. During the experiment, the animals' weights and tumor sizes were measured twice a week, and their clinical symptoms were observed and recorded daily. Each administration was based on the most recent weight measurement of the animals.

[0093] The evaluation index of antitumor activity was the relative tumor growth rate (T / C) (%). T / C (%) > 40% was ineffective, and T / C (%) ≦ 40% was effective after statistical processing with P < 0.05. The calculation formula for T / C (%) was T / C (%) = (T RTV / C RTV ) × 100%. T RTV is the relative tumor volume in the treatment group, and C RTV is the relative tumor volume of the negative control group, and TGI (%) bTumor inhibition rate = (mean tumor volume or weight of negative control group - mean tumor volume or weight of treatment group) / mean tumor volume or weight of negative control group x 100%. TGI (%) a = [(1 - (mean tumor volume at the time of measurement for a given treatment group - mean tumor volume at the start of treatment for that treatment group)) / (mean tumor volume at the time of measurement for the vehicle control group - mean tumor volume at the start of treatment for the vehicle control group)] × 100%.

[0094] Technical effects The compounds of the present invention have good inhibitory effects on porcupine protein, can effectively regulate Wnt / β-catenin pathway signaling, have good growth inhibitory effects on tumors overactivated by the Wnt / β-catenin signaling pathway, and have good pharmacokinetic properties.

[0095] Definitions and Explanations Unless otherwise explained, the following terms and phrases used herein are intended to have the following meanings. Certain terms or phrases, unless otherwise defined, should not be considered indefinite or unclear and should be understood according to their ordinary meaning. When trade names appear herein, it is intended to refer to the corresponding trade name or its active ingredient.

[0096] It should be noted that unless the context expressly dictates otherwise or is clearly contrary to the specification, the singular forms "a," "an," "said," and similar terms used in the present subject matter (particularly the subject matter of the appended claims) as used in this specification and the appended claims are to be construed as including both the singular and the plural. Thus, for example, reference to "said compound" includes reference to one or more compounds, etc.

[0097] As used herein, the term "pharmaceutically acceptable" refers to those compounds, materials, compositions and / or dosage forms which, within the scope of sound medical judgment, are suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response or other problem or complication, and are commensurate with a reasonable benefit / risk ratio.

[0098] The term "pharmaceutically acceptable salt" refers to a salt of a compound of the present invention prepared from a compound having certain substituents and a relatively non-toxic acid or base. When a compound of the present invention contains a relatively acidic functional group, a base addition salt can be obtained by contacting such a compound with a sufficient amount of base in a pure solution or in a suitable inert solvent. Pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amine, or magnesium salts or similar salts. When a compound of the present invention contains a relatively basic functional group, an acid addition salt can be obtained by contacting such a compound with a sufficient amount of acid in a pure solution or in a suitable inert solvent. Some specific compounds of the present invention contain both basic and acidic functional groups and can therefore be converted into either a base addition salt or an acid addition salt.

[0099] The pharmaceutically acceptable salts of the present invention can be synthesized from a parent compound that contains an acid or base group by conventional chemical methods. Generally, such salts are prepared by reacting the free acid or free base form of these compounds with a stoichiometric amount of the appropriate base or acid in water or an organic solvent, or a mixture of both.

[0100] The compounds of the present invention may exist in specific geometric or stereoisomeric forms. All such compounds contemplated by the present invention include cis and trans isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers, (D)-isomers, (L)-isomers, and racemic and other mixtures thereof, such as enantiomer- or diastereomer-enriched mixtures, all of which are within the scope of the present invention. Other asymmetric carbon atoms may be present in substituents such as alkyl. All of these isomers and mixtures thereof are within the scope of the present invention.

[0101] Unless otherwise specified, the terms "enantiomers" or "optical isomers" refer to stereoisomers that are mirror images of each other.

[0102] Unless otherwise stated, the terms "cis-trans isomers" or "geometric isomers" refer to the inability to freely rotate about double bonds or single bonds of ring carbon atoms.

[0103] Unless otherwise stated, the term "diastereomer" refers to stereoisomers whose molecules have two or more centers of chirality and whose molecules are not mirror images of each other.

[0104] Unless otherwise stated, "(+)" means dextrorotatory, "(-)" means levorotatory, and "(±)" means racemic.

[0105] Unless otherwise stated, [ka] Represents.

[0106] The compounds of the present invention may exist in specific forms. Unless otherwise specified, the term "tautomer" or "tautomeric form" refers to isomers of different functional groups that are in dynamic equilibrium at room temperature and are rapidly interconvertible. If tautomers are possible (e.g., in solution), chemical equilibrium of the tautomers can be achieved. For example, proton tautomers (also called prototropic tautomers) include interconversions via the migration of a proton, such as keto-enol isomerization and imine-enol isomerization. Valence tautomers include interconversions via recombination of some bond electrons. A specific example of keto-enol tautomerization is the interconversion between the two tautomers of pentane-2,4-dione and 4-hydroxypent-3-en-2-one.

[0107] Unless otherwise specified, the terms "enriched in one isomer," "enriched in an isomer," "enriched in one enantiomer," or "enantiomer-enriched" refer to a mixture that contains less than 100% of one isomer or enantiomer, and that contains at least 60%, or at least 70%, or at least 80%, or at least 90%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99%, or at least 99.5%, or at least 99.6%, or at least 99.7%, or at least 99.8%, or at least 99.9% of that isomer or enantiomer.

[0108] Unless otherwise stated, the terms "isomeric excess" or "enantiomeric excess" refer to the difference between the relative percentages of two isomers or two enantiomers. For example, if one isomer or enantiomer is present at 90% and the other isomer or enantiomer is present at 10%, the isomeric or enantiomeric excess (ee) is 80%.

[0109] Optically active (R)- and (S)-isomers, as well as D- and L-isomers, can be prepared by chiral synthesis or chiral reagents or other conventional techniques. Single enantiomers of certain compounds of the invention can be prepared by asymmetric synthesis or derivatization with a chiral auxiliary, where the resulting diastereomeric mixture can be separated and the auxiliary cleaved to provide the pure desired enantiomer. Alternatively, if the molecule contains a basic (e.g., amino) or acidic (e.g., carboxyl) functional group, the diastereomeric salt can be formed with an appropriate optically active acid or base, followed by separation and recovery of the diastereomers by conventional methods known in the art to provide the pure enantiomers. Separation of enantiomers and diastereomers is also typically carried out using chromatography on chiral stationary phases, optionally in combination with chemical derivatization (e.g., carbamate formation from amines). The compounds of the invention may contain unnatural proportions of atomic isotopes at one or more of the atoms that constitute the compounds. For example, compounds containing tritium ( 3 H), iodine-125(125 I), or C-14( 14 The compounds of the present invention can be labeled with a radioactive isotope such as HCl, HCl, or HCl. Alternatively, for example, deuterium can be substituted for hydrogen to form a deuterated drug, where the bond formed between deuterium and carbon is stronger than the bond formed between normal hydrogen and carbon. Compared with non-deuterated drugs, deuterated drugs have the advantages of reduced toxic side effects, increased drug stability, enhanced efficacy, and prolonged biological half-life. Conversion of the isotopic composition of the compounds of the present invention, whether radioactive or not, is included within the scope of the present invention.

[0110] The term "optionally" or "optionally" means that the subsequently described event or circumstance may, but need not, occur, and the description includes instances where the event or circumstance occurs and instances where the event or circumstance does not occur.

[0111] The term "substituted" means that any one or more hydrogen atoms at a particular atom are replaced with a substituent, and the substituent may include variants of deuterium and hydrogen, as long as the valence of the particular atom is normal and the substituted compound is stable. When the substituent is oxygen (i.e., =O), it means that two hydrogen atoms are replaced. Oxygen substitution does not occur in aromatic groups. The term "optionally substituted" means that the group may or may not be substituted, and unless otherwise specified, the type and number of substituents are optional as long as they are chemically feasible.

[0112] When any variable (e.g., R) occurs more than one time in any composition or structure of a compound, its definition is independent at each occurrence. Thus, for example, if a group is substituted with 0 to 2 R, then that group may optionally be substituted with up to 2 R, and each occurrence of R has independent options. Also, combinations of substituents and / or variants thereof are permissible only if such combinations result in stable compounds.

[0113] When the number of linking groups is 0, for example, -(CRR)0- means that the linking group is a single bond.

[0114] When one of the variables is a single bond, it means that the two groups linked to it are directly linked; for example, when L in ALZ represents a single bond, it means that the structure is actually AZ.

[0115] When a substituent is empty, it means that the substituent is not present, for example, when X in AX is empty, it means that the structure is actually A. When a recited substituent does not indicate through which atom it is bonded to the substituted group, such substituent can be bonded through any atom thereof, for example, pyridinyl as a substituent may be bonded to the substituted group through any carbon atom of the pyridine ring.

[0116] When the listed linking group does not indicate the direction of the link, the direction of the link is arbitrary, for example, [ka] In the formula, the linking group L is -MW-, and in this case, -MW- connects ring A and ring B in the same direction as the reading order from left to right. [ka] and by concatenating ring A and ring B in the reverse order of reading from left to right, [ka] Combinations of the above linking groups, substituents and / or variables thereof are permissible only if such combinations result in stable compounds.

[0117] Unless otherwise specified, when a group has one or more bondable sites, any one or more sites of the group can be bonded to other groups via a chemical bond. If the bonding mode of the chemical bond is delocalized and there is an H atom at the bondable site, when bonding with a chemical bond, the number of H atoms at the site is reduced to the group with the corresponding valence according to the number of bonded chemical bonds. The chemical bond that bonds the site to another group is [ka] For example, the straight solid bond in -OCH3 represents a bond to another group via the oxygen atom in the group, [ka] When one chemical bond is bonded, the H at that site decreases by one to the corresponding monovalent piperidinyl.

[0118] Unless otherwise specified, the number of atoms in a ring is generally defined as the number of ring members, e.g., a "5- to 7-membered ring" refers to a "ring" of 5 to 7 atoms arranged around it.

[0119] Unless otherwise stated, "C 1-3 The term "alkyl" is used to refer to a straight or branched chain saturated hydrocarbon group of 1 to 3 carbon atoms. 1-3 C for alkyl 1-2 and C 2-3 alkyl, which may be monovalent (e.g., methyl), divalent (e.g., methylene), and polyvalent (e.g., methine). 1-3 Examples of alkyl include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), and the like.

[0120] Unless otherwise stated, "C 1-3 The term "alkoxy" means an alkyl group containing 1 to 3 carbon atoms attached to the remainder of the molecule through an oxygen atom. 1-3 Alkoxy includes C1-2 , C 2-3 , C3 and C2 alkoxy, etc. 1-3 Illustrative examples of alkoxy include, but are not limited to, methoxy, ethoxy, propoxy (including n-propoxy or isopropoxy), and the like.

[0121] Unless otherwise stated, "C 1-3 The term "alkylamino" means an alkyl group containing 1 to 3 carbon atoms attached to the rest of the molecule through an ammonia atom. 1-3 Alkylamino has C 1-2 , C3 and C2 alkylamino, etc. 1-3 Illustrative examples of alkylamino include, but are not limited to, -NHCH3, -N(CH3)2, -NHCH2CH3, -N(CH3)CH2CH3, -NHCH2CH2CH3, -NHCH2(CH3)2, and the like.

[0122] Unless otherwise stated, in the present invention, the terms "9- to 10-membered heteroaryl ring" and "9- to 10-membered heteroaryl" can be used interchangeably, and the term "9- to 10-membered heteroaryl" refers to a cyclic group having a conjugated π-electron system consisting of 9 to 10 ring atoms, of which 1, 2, 3, or 4 ring atoms are heteroatoms independently selected from O, S, and N, and the remainder are carbon atoms. It may be a monocyclic, fused bicyclic, or fused tricyclic ring system, in which each ring is aromatic, wherein the nitrogen atom is optionally quaternized, and the nitrogen and sulfur heteroatoms may optionally be oxidized (i.e., NO and S(O)). p, p is 1 or 2). The 5- to 10-membered heteroaryl can be attached to the remainder of the molecule via a heteroatom or a carbon atom. The 9- to 10-membered heteroaryl includes 9-membered and 10-membered heteroaryls. Illustrative examples of the 9- to 10-membered heteroaryl include, but are not limited to, benzothiazolyl (including 5-benzothiazolyl, etc.), purinyl, benzimidazolyl (including 2-benzimidazolyl, etc.), benzoxazolyl, indolyl (including 5-indolyl, etc.), isoquinolyl (including 1-isoquinolyl and 5-isoquinolyl, etc.), quinoxalinyl (including 2-quinoxalinyl and 5-quinoxalinyl, etc.), or quinolyl (including 3-quinolyl and 6-quinolyl, etc.).

[0123] Unless otherwise stated, C n-n+m or C n ~C n+m includes any one specific embodiment of n to n+m carbons, for example, C 1-12 are C1, C2, C3, C4, C5, C6, C7, C8, C9, C 10 , C 11 , and C 12 and any one of the ranges n to n+m, for example, C 1-12 is C 1-3 , C 1-6 , C 1-9 , C 3-6 , C 3-9 , C 3-12 , C 6-9 , C 6-12 , and C 9-12 Similarly, n- to n+m-membered rings indicate that the number of atoms in the ring is n to n+m, and for example, a 3- to 12-membered ring includes a 3-membered ring, a 4-membered ring, a 5-membered ring, a 6-membered ring, a 7-membered ring, an 8-membered ring, a 9-membered ring, a 10-membered ring, an 11-membered ring, and a 12-membered ring, and also includes any one range of n to n+m, and for example, a 3- to 12-membered ring includes a 3- to 6-membered ring, a 3- to 9-membered ring, a 5- to 6-membered ring, a 5- to 7-membered ring, a 6- to 7-membered ring, a 6- to 8-membered ring, and a 6- to 10-membered ring, etc.

[0124] The term "leaving group" refers to a functional group or atom that may be displaced by another functional group or atom via a substitution reaction (e.g., an affinity substitution reaction). For example, representative leaving groups include trifluoromethanesulfonate; chlorine, bromine, iodine; sulfonate groups such as methanesulfonate, toluenesulfonate, p-bromobenzenesulfonate, p-toluenesulfonate, etc.; acyloxy groups such as acetoxy, trifluoroacetoxy, etc.

[0125] The term "protecting group" includes, but is not limited to, an "amino-protecting group," a "hydroxy-protecting group," or a "mercapto-protecting group." The term "amino-protecting group" refers to a protecting group suitable for preventing side reactions at the amino nitrogen. Representative amino acid protecting groups include, but are not limited to, formyl; acyl such as alkanoyl (e.g., acetyl, trichloroacetyl, or trifluoroacetyl); alkoxycarbonyl such as tert-butoxycarbonyl (Boc); arylmethoxycarbonyl such as benzyloxycarbonyl (Cbz) and 9-fluorenylmethoxycarbonyl (Fmoc); arylmethyl such as benzyl (Bn), triphenylmethyl (Tr), 1,1-di-(4'-methoxyphenyl)methyl; silyl such as trimethylsilyl (TMS) and tert-butyldimethylsilyl (TBS). The term "hydroxyl-protecting group" refers to a protecting group suitable for preventing side reactions of hydroxyl. Representative hydroxyl protecting groups include, but are not limited to, alkyl such as methyl, ethyl, and tert-butyl; acyl such as alkanoyl (e.g., acetyl); arylmethyl such as benzyl (Bn), p-formyloxybenzyl (PMB), 9-fluorenylthyl (Fm) and diphenylmethyl (diphenylmethyl, DPM); silyl such as trimethylsilyl (TMS) and tert-butyldimethylsilyl (TBS); and the like.

[0126] The compounds of the present invention can be produced by various synthetic methods known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combining with other chemical synthetic methods, and equivalent alternative methods known to those skilled in the art, and preferred embodiments include, but are not limited to, the examples of the present invention.

[0127] Throughout this specification, references to "one embodiment" or "an embodiment" or "another embodiment" or "in some embodiments" mean that at least one embodiment includes the associated particular referenced element, structure, or feature described in that embodiment. Thus, the appearances of the phrases "in one embodiment" or "an embodiment" or "another embodiment" or "in some embodiments" in various places throughout this specification do not necessarily all refer to the same embodiment. Furthermore, particular elements, structures, or features may be combined in any suitable manner in one or more embodiments.

[0128] Differential scanning calorimetry (DSC) of the crystals described in the present invention has experimental errors and is slightly affected by the degree of drying of the sample, and the position and value of the endothermic peak may differ slightly between one machine and another and between one sample and another, and the experimental error or difference may be 10°C or less, or 9°C or less, or 8°C or less, or 7°C or less, or 6°C or less, or 5°C or less, or 4°C or less, or 3°C or less, or 2°C or less, or 1°C or less, so the peak position or peak value of the DSC endothermic peak cannot be considered absolute.

[0129] The structure of the compound of the present invention can be confirmed by conventional methods known to those skilled in the art. When the present invention relates to the absolute configuration of a compound, the absolute configuration can be confirmed by conventional technical means of those skilled in the art. For example, in single crystal X-ray diffraction (SXRD), the grown single crystal is collected by a Bruker D8 venture diffractometer to collect diffraction intensity data, the light source is CuKα radiation, and the scanning method is φ / ω scanning. After collecting the relevant data, the crystal structure can be analyzed by a direct method (Shelxs97) to confirm the absolute configuration.

[0130] The present invention will be specifically explained below with reference to examples, but these examples do not limit the present invention in any way.

[0131] The solvents used in the present invention can be obtained commercially.

[0132] The following abbreviations are used in the present invention: DMF stands for N,N-dimethylformamide; K2CO3 stands for potassium carbonate; Cs2CO3 stands for cesium carbonate; EtOAc stands for ethyl acetate; EA stands for ethyl acetate; THF stands for tetrahydrofuran; MeOH stands for methanol; DCM stands for dichloromethane; DMSO stands for dimethyl sulfoxide; PE stands for petroleum ether; EtOH stands for ethanol; ACN stands for acetonitrile; TFA stands for trifluoroacetic acid; FA stands for formic acid; NH3·H2O stands for aqueous ammonia; TEA stands for triethylamine; DIPEA stands for N,N-diisopropylethylamine; Boc2O stands for di-tert-butyl dicarbonate; Boc stands for tert-butoxycarbonyl, an amino protecting group; LCMS stands for liquid chromatography mass spectrometry; HPLC stands for high performance liquid chromatography; TLC stands for thin layer chromatography; SFC stands for supercritical fluid chromatography; g stands for grams; mg stands for milligrams. μL stands for microliter. mL stands for milliliter. mol stands for mole. mmol stands for millimole. μmol stands for micromole. M stands for moles / liter. mM stands for millimoles / liter. μM stands for micromoles / liter. nM stands for nanomoles / liter. Me stands for methyl. Boc stands for tert-butoxycarbonyl. DMSO-d6 stands for deuterated dimethyl sulfoxide. CD3OD-d4 stands for deuterated methanol. CDCl3 stands for deuterated chloroform.

[0133] Compounds are named according to conventional naming principles in the art or using ChemDraw® software, commercially available compounds are named in supplier catalogs.

[0134] The test parameters for the X-ray powder diffractometer (XRPD) method of the present invention are as shown in Table 3.

[0135] [Table 3] The test parameters for the Differential Scanning Calorimeter (DSC) method of the present invention are as shown in Table 4.

[0136] [Table 4] The test parameters for the thermal gravimetric analyzer (TGA) method of the present invention are as shown in Table 5.

[0137] [Table 5] The test parameters for the Dynamic Vapor Sorption (DVS) method of the present invention are as shown in Table 6.

[0138] [Table 6] [Brief explanation of the drawings]

[0139] [Figure 1] 1 is an XRPD pattern of Cu-Kα radiation of the A-type crystal of Compound 2. [Figure 2] 1 is a DSC pattern of type A crystals of Compound 2. [Figure 3] 1 is a TGA pattern of type A crystals of Compound 2. [Figure 4] 1 shows the DVS pattern of type A crystals of Compound 2. [Figure 5] 1 is an XRPD pattern of Cu-Kα radiation of B-type crystals of Compound 2. [Figure 6] 1 is a DSC pattern of B-type crystals of Compound 2. [Figure 7] 1 is a TGA pattern of type B crystals of Compound 2. DETAILED DESCRIPTION OF THE INVENTION

[0140] The present invention will be described in detail below with reference to examples, which should not be construed as limiting the present invention in any way. Although the present invention has been described in detail herein, specific embodiments thereof have also been disclosed, and it will be apparent to those skilled in the art that various changes and modifications can be made to the specific embodiments of the present invention without departing from the spirit and scope of the present invention.

[0141] [ka] [ka] Step 1 Compound A-1 (15 g, 77.95 mmol) was dissolved in tetrahydrofuran (150 mL), and N-formylmorpholine (11.67 g, 101.33 mmol) and isopropyl magnesium chloride-lithium chloride (1.3 M, 89.94 mL) were added to the reaction solution. The reaction mixture was purged with gas and stirred at 25 ° C. under nitrogen gas protection for 2 hours. After the reaction was completed, the mixture was diluted with water (300 mL) and extracted with ethyl acetate (100 mL × 3). The combined organic phase was washed with saturated sodium chloride solution (200 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain crude product A-2.

[0142] Step 2 Compound A-2 (11 g, 77.71 mmol) was dissolved in tetrahydrofuran (150 mL). tert-butylsulfinamide (18.84 g, 155.42 mmol), sodium sulfate (5.52 g, 38.85 mmol), and copper sulfate (37.21 g, 233.12 mmol) were added to the reaction solution. The reaction mixture was stirred at 80 °C for 12 hours. After the reaction was completed, it was filtered, and the filtrate was diluted with water (200 mL) and extracted with ethyl acetate (150 mL × 3). The combined organic phase was washed with saturated sodium chloride solution (300 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain crude product A-3. MS-ESI calculated value [M + H] + 245, actual value 245.

[0143] Step 3 Compound A-3 (19 g, 77.63 mmol) was dissolved in tetrahydrofuran (200 mL). Sodium borohydride (1.81 g, 47.85 mmol) was added to the reaction solution, and the reaction mixture was stirred at 25 °C for 2 hours. After the reaction was completed, the mixture was diluted with water (350 mL) and extracted with ethyl acetate (150 mL × 3). The combined organic phase was washed with saturated sodium chloride solution (300 mL), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 100:1 to 1:1) to obtain compound A-4. MS-ESI calculated value [M + H] + 247, actual value 247.

[0144] Step 4 Compound A-4 (1.38 g, 5.61 mmol) was dissolved in dioxane (20 mL) and water (10 mL). Compound A-5 (1.45 g, 6.17 mmol), potassium carbonate (1.55 g, 11.22 mmol), and [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (205.14 mg, 280.45 μmol) were added to the reaction solution, and the reaction mixture was stirred at 80 ° C for 4 hours. After the reaction was completed, the mixture was filtered, concentrated under reduced pressure, and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 10 / 1 to 1 / 1) to obtain compound A-6. MS-ESI calculated value [M + H] + 320, actual measured value 320.

[0145] Step 5 Compound A-6 (2.8 g, 8.77 mmol) was dissolved in methanol (20 mL), and hydrogen chloride / ethyl acetate (4 M, 10.96 mL) was added to the reaction solution. The reaction mixture was stirred at 25°C for 4 hours. After the reaction was completed, the mixture was filtered, and the cake was dried under vacuum to obtain the hydrochloride salt of intermediate A. MS-ESI calculated values ​​[M+H] + 216, actual value 216.

[0146] Example 1 [ka] Step 1 The hydrochloride salt of intermediate A (200 mg) and compound 1-1 (118.67 mg, 554.47 μmol) were dissolved in N,N-dimethylformamide (3 mL). N,N-dimethylisopropylamine (636.99 mg, 4.93 mmol) and benzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate (35.97 mg, 985.73 μmol) were added to the reaction solution, and the reaction mixture was stirred at 50 °C for 12 h. After the reaction was completed, the mixture was diluted with water (5 mL) and extracted with ethyl acetate (10 mL x 3). The combined organic phase was washed with saturated sodium chloride solution (10 mL x 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product, compound 1-2, was obtained by silica gel column chromatography (eluent: dichloromethane / methanol, 1 / 0 to 10 / 1, v / v). MS-ESI calculated values ​​[M+H]. + 412, actual value 412.

[0147] Step 2 Compound 1-2 (80 mg, 194.53 μmol), compound 1-3 (54.82 mg, 389.05 μmol), and potassium phosphate (123.87 mg, 583.58 μmol) were dissolved in tetrahydrofuran (2 mL) and water (0.5 mL). The reaction mixture was purged with nitrogen gas three times. Under nitrogen gas protection, [1,1'-bis(di-tert-butylphosphino)ferrocene]palladium(II) dichloride (25.36 mg, 38.91 μmol) was added and stirred at 70 °C for 12 hours under nitrogen gas protection. After the reaction was completed, the mixture was filtered, the filtrate was concentrated under reduced pressure, and then diluted with water (5 mL). The mixture was extracted with dichloromethane:methanol (10:1, V / V, 10 mL × 3). The combined organic phase was washed with saturated sodium chloride solution (10 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was first separated and purified by thin-layer chromatography (eluent: dichloromethane / methanol, 10 / 1, V / V) to obtain a crude product, which was then purified by high-performance liquid chromatography (chromatography column: Phenomene×luna C18 150 × 40 mm × 15 μm, mobile phase: mobile phase A: 0.225% by volume aqueous formic acid, mobile phase B: acetonitrile, B%: 28%-55%) to obtain compound 1.

[0148] MS-ESI calculated value [M+H] + 428, actual value 428. 1 H NMR (400 MHz, DMSO-d6) δ ppm 9.16 (t, J = 1.6 Hz, 1 H), 9.01 (t, J = 5.8 Hz, 1 H), 8.61 (d, J = 1.4 Hz, 1 H) 8.45 - 8.54 (m, 3 H), 8.10 - 8.18 (m, 2 H), 7.75 - 7.85 (m, 2 H), 7.33 (d, J = 4.6 Hz, 1 H), 7.12 (d, J = 4.6 Hz, 1 H), 6.49 (d, J = 9.4 Hz, 1 H), 4.80 (d, J = 5.8 Hz, 2 H), 3.53 (s, 3H).

[0149] Example 2 [ka] Step 1 Compound 2-1 (7.9 g, 29.53 mmol), compound 2-2 (4.4 g, 29.53 mmol), and cesium carbonate (9.62 g, 29.53 mmol) were dissolved in N,N-dimethylformamide (44 mL). The reaction mixture was purged with nitrogen gas three times and stirred at 20 °C under nitrogen gas protection for 21 h. After the reaction was complete, the mixture was diluted with water (50 mL) and extracted with ethyl acetate (100 mL × 3). The combined organic phase was washed with saturated sodium chloride solution (50 mL × 2), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate, 1 / 0 to 20 / 1, v / v) to obtain crude product 2-3. MS-ESI calculated values ​​[M+H] + 380, actual value 380.

[0150] Step 2 Compound 2-3 (10 g, 26.33 mmol) was dissolved in water (9.1 mL) and toluene (27 mL), and hydrochloric acid (2.67 g, 26.33 mmol, purity: 36%) was added. The mixture was reacted at 25°C for 21 hours. After the reaction was completed, the mixture was extracted with toluene (20 mL x 3) and the aqueous phases were combined. Next, concentrated hydrochloric acid (2.36 mL) was added to the aqueous phase, and the mixture was reacted at 60°C for 20 hours. After the reaction was completed, the reaction solution was concentrated under reduced pressure to obtain the hydrochloride salt of compound 2-4. MS-ESI calculated value [M+H] + 144, measured value 144.

[0151] Step 3 The hydrochloride salt of compound 2-4 (3.8 g) and sodium bicarbonate (22.23 g, 264.67 mmol) were dissolved in dichloromethane (35 mL) and acetonitrile (35 mL). Di-tert-butyl dicarbonate (8.66 g, 39.70 mmol) was added, and the mixture was reacted at 25 °C for 16 hours. After the reaction was completed, the mixture was diluted with water (100 mL) and extracted with ethyl acetate (100 mL × 3). The combined organic phase was washed with saturated sodium chloride solution (50 mL × 2), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate, 1 / 0 to 10 / 1, v / v) to obtain crude product 2-5. MS-ESI calculated values ​​[M+H] + 244, actual value 244.

[0152] Step 4 Compound 2-5 (3 g, 12.31 mmol), compound A-5 (3.47 g, 14.77 mmol), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (450.39 mg, 615.54 μmol), and potassium carbonate (3.40 g, 24.62 mmol) were dissolved in water (3 mL) and dioxane (30 mL). The reaction mixture was purged with nitrogen gas three times and stirred at 80 °C for 12 h under nitrogen gas protection. After the reaction was complete, the mixture was diluted with water (50 mL) and extracted with ethyl acetate (100 mL × 2). The combined organic phase was washed with saturated sodium chloride solution (50 mL × 2), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by silica gel column chromatography (eluent: dichloromethane / methanol, 1 / 0 to 50 / 1, v / v) to obtain crude product 2-6. MS-ESI calculated values ​​[M+H] + 317, measured value 317.

[0153] Step 5 Compound 2-6 (5.64 g, 17.83 mmol) was dissolved in methanol (50 mL), and hydrogen chloride / ethyl acetate (44.57 mL, 178.3 mmol, 4 M) was added. The mixture was reacted at 25°C for 12 hours. After the reaction was completed, the reaction solution was concentrated under reduced pressure to obtain a crude product. Ethyl acetate:methanol (55 mL, 10 / 1, V / V) was added to the crude product, and the mixture was stirred at 25°C for 1 hour. The product was then filtered to obtain a cake, which was then concentrated under reduced pressure to obtain the hydrochloride salts of compounds 2-7. MS-ESI calculated values ​​[M+H] + 217, actual value 217.

[0154] Step 6 The hydrochloride salt (1 g) of compound 2-7 was dissolved in N-methylpyrrolidone (10 mL), and N,N-dimethylisopropylamine (1.8 g, 13.96 mmol) and compound 2-8 (584.11 mg, 2.51 mmol) were added. The reaction solution was reacted at 130 °C for 16 hours. After the reaction was completed, the mixture was diluted with water (50 mL) and filtered to obtain a cake. The cake was concentrated under reduced pressure to obtain compound 2-9. MS-ESI calculated values ​​[M+H] + 413, actual value 413.

[0155] Step 7 Compound 2-9 (400 mg, 970.3 μmol), compound 2-10 (229.83 mg, 1.46 mmol), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (35.5 mg, 48.52 μmol), and potassium carbonate (268.2 mg, 1.94 mmol) were dissolved in dioxane (4 mL) and water (0.8 mL). The reaction mixture was purged with nitrogen gas three times and stirred at 80 °C under nitrogen gas protection for 12 hours. After completion of the reaction, the mixture was diluted with water (20 mL) and extracted with ethyl acetate (50 mL × 3). The combined organic phase was washed with saturated sodium chloride solution (20 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was first purified by silica gel column chromatography (eluent: dichloromethane / methanol, 1 / 0 to 20 / 1, V / V) to obtain a crude product, and then purified by high-performance liquid chromatography (chromatography column: Phenomene×Gemini-N×C18 75×30 mm×15 μm, mobile phase: mobile phase A: 0.05% aqueous ammonia solution by volume, mobile phase B: acetonitrile, B%: 18% to 48%) to obtain compound 2.

[0156] MS-ESI calculated value [M+H] + 446, actual value 446. 1 H NMR (400 MHz, DMSO-d6) δ ppm 9.03 (d, J = 1.4 Hz, 1 H), 8.59 (m, 2 H), 8.28 (m, 1 H) 8.16 (m, 1 H), 7.85 - 7.95 (m, 2 H), 7.47 (m, 2 H), 7.30 - 7.40 (m, 2 H), 6.53 (d, J = 9.6 Hz, 1 H), 4.85 (d, J = 6.0 Hz, 2 H), 3.54 (s, 3 H).

[0157] Experimental Example 3 [ka] Compound 2-9 (400 mg, 970.3 μmol), compound 1-3 (205.08 mg, 1.46 mmol), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (25.5 mg, 48.52 μmol), and potassium carbonate (268.2 mg, 1.94 mmol) were dissolved in dioxane (4 mL) and water (0.8 mL). The reaction mixture was purged with nitrogen gas three times, and the mixture was stirred under nitrogen gas protection until it was heated to 1000°C. [ert-butylphosphino]ferrocenepalladium(II) dichloride (25.36 mg, 38.91 μmol) was added and stirred at 80 °C for 12 h under nitrogen gas protection. After completion of the reaction, the mixture was filtered, the filtrate was concentrated under reduced pressure, diluted with water (20 mL), extracted with ethyl acetate (50 mL × 3), the combined organic phase was washed with saturated sodium chloride solution (20 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was first purified by silica gel column chromatography (eluent: dichloromethane / methanol, 1 / 0 to 20 / 1, v / v) to obtain the crude product. The crude product was then purified by high-performance liquid chromatography (chromatography column: Waters Bridge C18 150 × 50 mm × 10 μm, mobile phase: mobile phase A: 10 mM aqueous ammonium bicarbonate solution, mobile phase B: acetonitrile, B%: 14% to 44%) to obtain compound 3.

[0158] MS-ESI calculated value [M+H] + 429, actual value 429. 1 H NMR (400 MHz, DMSO-d6) δ ppm 9.03 (d, J = 1.4 Hz, 1 H), 8.79 (s, 1 H), 8.67 (d, J = 2.6 Hz, 1 H), 8.60 (m, 2 H), 8.31 (m, 1 H), 8.09 - 8.20 (m, 2 H), 7.97 (s, 1 H), 7.91 (d, J = 4.8 Hz, 1 H), 7.34 (d, J = 4.8 Hz, 1 H), 6.53 (d, J = 9.6 Hz, 1 H), 4.85 (d, J = 5.8 Hz, 2 H), 3.54 (s, 3H).

[0159] Example 4 [ka] Step 1 The hydrochloride salt of compound intermediate A (500 mg) and compound 2-8 (539.99 mg, 2.32 mmol) were dissolved in N-methylpyrrolidone (15 mL). N,N-dimethylisopropylamine (300.21 mg, 2.32 mmol) was added to the reaction solution, and the reaction mixture was stirred at 130 °C for 16 hours. After the reaction was completed, the mixture was diluted with water (100 mL) and extracted with ethyl acetate (100 mL x 3). The combined organic phase was washed with saturated sodium chloride solution (100 mL x 3), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by silica gel column chromatography (petroleum ether:ethyl acetate:ethanol = 4:3:1) to obtain crude product 4-1. MS-ESI calculated values ​​[M+H] + 412, actual value 412.

[0160] Step 2 Compound 4-1 (955 mg, 2.32 mmol) and compound 1-3 (425.37 mg, 3.02 mmol) were dissolved in dioxane (10 mL) and water (2 mL). Potassium carbonate (962.81 mg, 6.97 mmol) and [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (169.91 mg, 232.22 μmol) were added to the reaction solution, and the reaction mixture was purged with gas and stirred at 110 °C for 16 hours under nitrogen gas protection. After the reaction was completed, the mixture was filtered, concentrated under reduced pressure, and purified by high-performance liquid chromatography (chromatography column: Phenomene×luna C18 150 × 40 mm × 15 μm, mobile phase: mobile phase A: 0.225% volume fraction formic acid aqueous solution, mobile phase B: acetonitrile, B%: 15% to 45%) to obtain compound 4.

[0161] MS-ESI calculated value [M+H] + 428, actual value 428. 1H NMR (400 MHz, DMSO-d6) δ ppm 8.77 (t, J = 1.6 Hz, 1 H), 8.66 (d, J = 2.8 Hz, 1 H), 8.27 (s, 1 H), 8.07 (d, J = 2.8 Hz, 1 H), 7.9 (s, 1 H), 7.86 (d, J = 4.8Hz, 1 H), 7.79 (d, J = 2.6 Hz, 1 H), 7.46 - 7.51 (m, 2 H), 7.38 - 7.43 (m, 2 H), 7.4 (d, J = 4.8 Hz, 1 H), 6.46 (d, J = 9.4 Hz, 1 H), 4.7 (d, J = 6.4 Hz, 2 H), 3.49 (s, 3 H).

[0162] Example 5 [ka] Step 1 Compound 5-1 (2 g, 13.02 mmol) was dissolved in N,N-dimethylformamide (10 mL). Compound 1-3 (2.39 g, 16.93 mmol), copper acetate (4.73 g, 26.05 mmol), and pyridine (3.09 g, 39.07 mmol) were added to the reaction solution. The reaction mixture was purged with gas and stirred at 40 °C under oxygen gas protection for 12 h. After completion of the reaction, water (100 mL) was added and the mixture was extracted with dichloromethane (100 mL). The combined organic phase was washed with saturated sodium chloride solution (25 mL × 2), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by silica gel column chromatography (petroleum ether:ethyl acetate = 10 / 1 to 2 / 1) to obtain compound 5-2. MS-ESI calculated values ​​[M+H]. + 249, actual value 249.

[0163] Step 2 Compound 5-2 (330 mg, 1.33 mmol) and the hydrochloride salt of intermediate A (428.52 mg) were dissolved in N-methylpyrrolidone (5 mL). N,N-dimethylisopropylamine (280.94 mg, 2.03 mmol) was added to the reaction solution, and the reaction mixture was stirred at 130 °C for 3 h. After the reaction was completed, the reaction solution was diluted with water (20 mL) and extracted with ethyl acetate (20 mL). The organic phase was washed with saturated sodium chloride solution (10 mL × 2) and purified by high-performance liquid chromatography (chromatography column: Phenomene × Luna C18 75 × 30 mm × 3 μm, mobile phase: mobile phase A: 0.225% formic acid aqueous solution by volume, mobile phase B: acetonitrile, B%: 1% to 30%) to obtain compound 5.

[0164] MS-ESI calculated value [M+H] + 428, actual value 428. 1 H NMR (400 MHz, DMSO-d6) δ ppm 8.84 (d, J = 1.6 Hz, 1 H), 8.75 (d, J = 2.4 Hz, 1 H), 8.59 (d, J = 1.6 Hz, 1 H), 8.56 (s, 1 H), 8.48 (d, J = 2.4 Hz, 1 H), 8.24 (m, 1 H), 8.12 (m, 1 H), 7.84 (d, J = 6.0 Hz, 1 H), 7.79 (m, 1 H), 7.73 (s, 1 H), 7.66 (t, J = 6.2 Hz, 1 H), 6.92 (d, J = 5.8 Hz, 1 H), 6.48 (d, J = 9.6 Hz, 1 H), 4.74 (d, J = 6.0 Hz, 2 H), 3.52 (s, 3 H).

[0165] Example 6 [ka] Step 1 Compound 6-1 (5 g, 26.87 mmol) was dissolved in dichloromethane (50 mL). Di-tert-butyl dicarbonate (7.04 g, 32.25 mmol) and N,N-dimethylisopropylamine (10.42 g, 80.62 mmol) were added to the reaction solution. The reaction mixture was purged with gas and stirred at 25 °C under nitrogen gas protection for 12 hours. After the reaction was completed, water (100 mL) was added and the mixture was extracted with dichloromethane (200 mL). The combined organic phase was washed with saturated sodium chloride solution (50 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give compound 6-2. MS-ESI calculated values ​​[M + H] + 287, actual value 287.

[0166] Step 2 Compound 6-2 (2 g, 6.99 mmol) and compound A-5 (2.05 g, 8.74 mmol) were dissolved in dioxane (30 mL) and water (3 mL). Potassium carbonate (2.90 g, 20.97 mmol) and [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (511.39 mg, 698.90 μmol) were added to the reaction solution, and the reaction mixture was stirred at 80 °C for 12 hours. After the reaction was completed, the mixture was filtered, concentrated under reduced pressure, and purified by silica gel column chromatography (petroleum ether / ethyl acetate = 5 / 1 to 0 / 1) to obtain compound 6-3. MS-ESI calculated values ​​[M+H] + 315, actual value 315.

[0167] Step 3 Compound 6-3 (2 g, 6.36 mmol) was dissolved in ethyl acetate (40 mL), and hydrogen chloride / ethyl acetate (4 M, 7.95 mL) was added to the reaction solution. The reaction mixture was purged with gas and stirred at 25°C under nitrogen gas protection for 12 hours. After the reaction was completed, the mixture was filtered, and the cake was dried in vacuo to obtain the hydrochloride salt of compound 6-4.

[0168] Step 4 Compound 6-4 hydrochloride (2 g) and compound 2-8 (1.84 g, 7.93 mmol) were dissolved in N-methylpyrrolidone (20 mL). N,N-dimethylisopropylamine (6.03 g, 46.67 mmol) was added to the reaction solution. The reaction mixture was purged with gas and stirred at 130 °C under nitrogen gas protection for 12 hours. After the reaction was complete, the mixture was diluted with water (100 mL) and extracted with dichloromethane (100 mL). The combined organic phase was washed with saturated sodium chloride solution (100 mL), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by silica gel column chromatography (petroleum ether / ethyl acetate = 1 / 1 to ethyl acetate / methanol = 20 / 1) to obtain compound 6-5. MS-ESI calculated values ​​[M+H] + 411, measured value 411.

[0169] Step 5 Compound 6-5 (500 mg, 1.22 mmol) and compound 1-3 (206.07 mg, 1.46 mmol) were dissolved in dioxane (10 mL) and water (2 mL). Potassium carbonate (505.30 mg, 3.66 mmol) and [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (89.17 mg, 121.87 μmol) were added to the reaction solution. The reaction mixture was purged with gas and stirred at 130 °C for 12 hours under nitrogen gas protection. After the reaction was completed, the mixture was filtered, concentrated under reduced pressure, and purified by high-performance liquid chromatography (chromatography column: Phenomene×luna C18 75 × 30 mm × 3 μm, mobile phase: mobile phase A: 0.225% volume fraction formic acid aqueous solution, mobile phase B: acetonitrile, B%: 22% to 52%) to obtain compound 6.

[0170] MS-ESI calculated value [M+H] + 427, actual value 427. 1H NMR (400 MHz, DMSO-d6) δ ppm 8.77 (t, J = 1.6 Hz, 1 H), 8.66 (d, J = 2.8 Hz, 1 H), 8.27 (s, 1 H), 8.09 - 8.13 (m, 1 H), 8.07 (d, J = 2.8 Hz, 1 H), 7.93 (s, 1 H), 7.86 (d, J = 4.8 Hz, 1 H), 7.79 (d, J = 2.6 Hz, 1 H), 7.46 - 7.51 (m, 2 H), 7.38 - 7.43 (m, 2 H), 7.34 (d, J = 4.8 Hz, 1 H), 6.46 (d, J = 9.4 Hz, 1 H), 4.71 (d, J = 6.4 Hz, 2 H), 3.49 (s, 3 H).

[0171] Example 7 [ka] Step 1 Compound 7-1 (1.5 g, 9.77 mmol) was dissolved in N,N-dimethylformamide (10 mL), followed by the addition of phenylboronic acid (2.38 g, 19.54 mmol), copper acetate (3.55 g, 19.54 mmol), and pyridine (1.55 g, 19.54 mmol, 1.58 mL). The mixture was stirred under oxygen gas at 25°C for 12 hours. After the reaction was completed, aqueous ammonia (20 mL) and ethyl acetate (50 mL) were added, followed by stirring at 25°C for 50 minutes. Water (30 mL) was added to the mixture, followed by extraction with ethyl acetate (50 mL x 3). The organic phase was washed with saturated brine (30 mL x 2), collected, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give compound 7-2. MS-ESI calculated values ​​[M+H] + 230, actual value 230.

[0172] Step 2 Compound 7-2 (200 mg, 870.83 μmol), the hydrochloride salt of compound intermediate A (339.24 mg), and 1,8-diazabicyclo[5.4.0]undec-7-ene (530.29 mg, 3.48 μmol) were dissolved in N-methylpyrrolidone (3 mL). After the reaction was completed, water (20 mL) was added and the mixture was extracted with ethyl acetate (20 mL × 3). The combined organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. The concentrate was purified by thin-layer chromatography (eluent: dichloromethane / methanol = 10 / 1) to obtain a crude product. The crude product was separated and purified by high-performance liquid chromatography (chromatography column: Waters Xbridge C18 150 × 50 mm × 10 μm, mobile phase: mobile phase A: 10 mM ammonium bicarbonate aqueous solution, mobile phase B: acetonitrile, B%: 20% to 50%) to obtain compound 7.

[0173] MS-ESI calculated value [M+H] + 409, actual value 409. 1 H NMR (400 MHz, CD3OD) δ 8.66 ppm (d, J = 1.8 Hz, 1H), 8.38 (d, J = 2.4 Hz, 1H), 8.35 (s, 1H), 8.19 (m, 1H), 7.92 (m, 1H), 7.84 (d, J = 6.2 Hz, 1H), 7.73 (d, J = 8.2 Hz, 1H), 7.69-7.60 (m, 4H), 7.59-7.53 (m, 1H), 6.92 (d, J = 6.2 Hz, 1H), 6.66 (d, J = 9.4 Hz, 1H), 4.88-4.87 (m, 2H), 3.68 (s, 3H).

[0174] Example 8: Preparation of Type A Crystals of Compound 2 100 mg of the compound of formula 2 was weighed and placed in a 4.0 ml glass vial, and an appropriate amount of methanol was added to form a suspension. After adding a magnet, the suspension sample was placed on a magnetic heating stirrer (25 °C) and stirred overnight at 25 °C. After filtration, the solid cake was dried overnight in a vacuum drying oven (45 °C) to obtain type A crystals of compound 2. The XRPD, DSC, TGA, and DVS detection results for type A crystals of compound 2 are shown in Figures 1, 2, 3, and 4, respectively.

[0175] Example 9: Preparation of B-type crystals of Compound 2 100 mg of the A-type crystals of the compound of formula 2 were weighed and added to a micro vial, which was then placed in a muffle furnace and heated at 260° C. for 5 minutes to obtain a B-type crystal compound.

[0176] Example 10: Solvent Pre-stabilization Study of Crystalline Form A of Compound 2 100 mg of compound 2 type A crystals were weighed and placed in a 4.0 ml glass vial, and an appropriate amount of solvent was added to form a suspension (during the experiment, the amount of compound and solvent was adjusted according to the test phenomenon, or the container used for the experiment was replaced, in order to suspend the sample as much as possible). After adding a magnet, the above suspension sample was placed on a magnetic heating stirrer (25 ° C) to perform the test. After stirring at 25 ° C overnight, it was filtered, and the solid sample on the cake was dried in a vacuum drying oven (45 ° C) overnight. The resulting solid crystals were detected by (XRPD). The test results are shown in Table 7.

[0177] [Table 7] Conclusion: Form A crystal of compound 2 had good stability in various solvents.

[0178] Example 11: Hygroscopicity study of Compound 2 Form A crystals Test materials: SMS DVS intrinsic dynamic water vapor adsorption measurement device Experimental Method: 10-30 mg of Compound 2 type A crystals were taken and placed in a DVS sample tray to perform the test.

[0179] The classification of moisture absorption evaluation is as shown in Table 8.

[0180] [Table 8] Note: ΔW% represents the moisture absorption weight gain of the test sample at 25±1°C and 80±2% RH.

[0181] Test Results: The DVS pattern of the A-type crystals of compound 2 is shown in Figure 4. The test sample was tested according to the 30-95% RH / 0-95% RH / 25°C procedure, with the sample pre-equilibrated at 30% RH. Compared to the initial humidity of 30%, as the humidity increased to 80%, the sample absorbed water, increasing in weight by 0.07% (cycle 1 adsorption). As the humidity continued to increase to 90%, the sample's weight decreased by 0.15%. As the humidity continued to increase to 95%, the sample absorbed water, increasing in weight by 0.01% (cycle 1 adsorption). The XRPD results showed that the diffraction peak positions of the sample before and after the DVS test were consistent, indicating that the crystal structure remained unchanged.

[0182] Testing Conclusion: The hygroscopic weight gain of the A-type crystals of Compound 2 at 25°C and 80% RH was 0.07%, indicating that the hygroscopicity was not or almost not significant.

[0183] Example 12: Preliminary solid-state stability test of Compound 2 type A crystals In accordance with the "Guiding Principles for Stability Testing of Drug Substances and Preparations" (Chinese Pharmacopoeia, 2015 Edition, Part IV, General Provisions 9001), the stability tests were conducted under high temperature (60°C, open), high humidity (room temperature / relative humidity 92.5%, open), and light irradiation (total irradiance = 1.2 × 10 6 Lux·hr / near ultraviolet=200w·hr / m 2 The stability of the A-type crystal of compound 2 under the conditions of condensation (open and closed) was investigated.

[0184] Ten milligrams of compound 2 type A crystals were weighed and placed at the bottom of a glass sample vial, allowing it to spread thinly. For samples exposed to high temperature (60°C) and high humidity (92.5% RH) conditions, the vial was sealed with aluminum foil, several small holes were poked in the foil to allow sufficient contact with the ambient air, and the sample was placed in a corresponding temperature and humidity oven. The irradiated sample (open, not covered with aluminum foil) and the irradiated control (the entire sample vial was covered with aluminum foil) were placed in the oven. Two portions were weighed at each time point as the official test sample. Approximately 50 mg of compound 2 type A crystals were also weighed and used for XRPD testing. The sample vial was wrapped in aluminum foil, small holes were poked in it, and the sample was similarly placed in a corresponding temperature and humidity oven. Samples were sampled on days 5 and 10 for XRPD analysis, and the results were compared with the initial results on day 0. The results of the solid state preliminary stability test of the A-type crystals of Compound 2 are shown in Table 9.

[0185] [Table 9] Conclusion: All of the A-type crystals of compound 2 had good stability under conditions of high temperature, high humidity and strong light irradiation.

[0186] Biological evaluation: Experimental Example 1: Super-Top-Flash (STF) reporter gene inhibitory activity test Test materials: HEK293 STF and L wnt3A cells were provided by Wuhan Heyan Biomedical Technology Co., Ltd., and Bright Glo was purchased from Promega.

[0187] Experimental Method: HEK293 STF cells and L wnt3A cells were seeded into a white 96-well plate at a 1:1 ratio, with 80 μL of cell suspension per well containing 20,000 HEK293 STF cells and 20,000 L wnt3A cells. The cell plate was cultured overnight in a carbon dioxide incubator.

[0188] The test compounds were diluted 5-fold to eight concentrations, from 400 μM to 5.12 nM, using a pipette. 78 μL of medium was added to the middle plate, and then 2 μL of the gradient-diluted compounds were transferred to the middle plate at the corresponding positions at each well. After uniform mixing, 20 μL of the compounds were transferred to the cell plate at each well. The cell plate was then incubated in a carbon dioxide incubator for 24 hours.

[0189] 100 μL of Promega Bright-Glo reagent was added per well to the cell plate and incubated at room temperature for 10 minutes to stabilize the luminescent signal. Data were read on a multilabel analyzer.

[0190] A multi-label analyzer was used to calculate raw data and IC values ​​were calculated through curve fitting. 50 The value can be obtained. Table 10 provides the inhibitory activity data of the compounds of the present invention against the Wnt signaling pathway. As a result, it was shown that the compounds of the present invention have good inhibitory activity against porcupine in the Wnt signaling pathway.

[0191] [Table 10] Experimental Example 2: Capan-2 cell proliferation inhibitory activity test Test materials: Capan-2 cells were provided by Wuhan Heyan Biomedical Technology Co., Ltd., and CellTiter-Glo was purchased from Promega.

[0192] Experimental Method: Capan-2 cells were seeded into a white 96-well plate, with 80 μL of cell suspension per well containing 5,000 Capan-2 cells, and the cell plate was cultured overnight in a carbon dioxide incubator.

[0193] Test compounds were diluted 3-fold to nine concentrations, from 200 μM to 30 nM, using a pipette. A replicate well experiment was set up. 78 μL of medium was added to the middle plate, and then 2 μL / well of the gradient-diluted compounds were transferred to the middle plate according to the corresponding positions. After uniform mixing, 20 μL / well of the compounds were transferred to the cell plate. The cell plate was placed in a carbon dioxide incubator and cultured for 5 days. A separate cell plate was prepared, and the signal value on the day of drug addition was taken as the maximum value (Max value in the equation below) and used for data analysis. 25 μL of cell viability chemiluminescence detection reagent was added to each well of this cell plate and incubated at room temperature for 10 minutes to stabilize the luminescence signal. The data were then read using a multilabel analyzer.

[0194] After culturing the cell plates for 5 days, 25 μL of Promega CellTiter-Glo reagent was added per well and incubated at room temperature for 10 minutes to stabilize the luminescence signal. Data were read using a multilabel analyzer.

[0195] I C 50 The values ​​can be obtained by curve fitting, and the results of the inhibitory activity of the compounds of the present invention against Capan-2 cell proliferation are shown in Table 11.

[0196] [Table 11] Experimental conclusion: The compounds of the present invention had good inhibitory activity against Capan-2 cell proliferation.

[0197] Experimental Example 3: In vivo efficacy study of HuPrime® gastric cancer GA3055 subcutaneous xenograft female BALB / C nude mouse model Test materials: GA3055 tumor-bearing mice were provided by Sino US Guanke Biotechnology (Taicang) Co., Ltd.

[0198] Experimental Method: Tumor tissue was collected from mice bearing the HuPrime® gastric cancer xenograft model GA3055, cut into tumor blocks 2-3 mm in diameter, and subcutaneously inoculated into the right anterior scapula of BALB / c nude mice. The average tumor volume of the tumor-bearing mice was approximately 150 mm. 3 When the mice reached 100 mg / kg, they were randomly assigned to groups. The coefficient of variation (CV) of tumor volume within each group was calculated using the formula CV = SD / MTV × 100% and was required to be less than 30%. Treatment was initiated on the day of group assignment (tumor volume was measured on the day of group assignment). The day of group assignment was defined as day 0.

[0199] Before the start of treatment, all animals were weighed and tumor volumes were measured using calipers. Because tumor volume affects the efficacy of treatment, mice were divided into groups according to tumor volume using a random grouping design method to ensure similar tumor volumes among different groups. Group division was performed using StudyDirector™ (version number: 3.1.399.19, supplier: Studylog System, Inc., San Francisco, CA, USA).

[0200] The "Matched distribution" random grouping method was used to divide the animals into groups, which can reflect the smallest intergroup differences in tumor volume levels. This algorithm compared the individual measurements of all selected animals with the mean value of all selected animals. First, paired animals whose mean value was close to the mean value of all selected animals were selected, and then the animals were assigned to groups so that the group mean value matched (or was as close as possible to) the mean value of all selected animals. The final mean value of the measurements of each group was as close as possible to the final mean value of the other groups.

[0201] Routine monitoring after tumor cell inoculation included tumor growth and the effects of treatment on the animals' normal behavior, including the activity of the experimental animals, food and water intake, weight gain and loss, eyes, hair coat, and other abnormalities. All clinical symptoms observed during the study were recorded in the raw data. After the start of treatment, the mice's weights and tumor sizes were measured twice a week. Tumor volume was calculated using the following formula: tumor volume (mm 3 )=1 / 2×(a×b 2 ) (a represents the long diameter and b represents the short diameter). In the experiment, StudyDirector™ (version number: 3.1.399.19, supplier: Studylog System, Inc.) software was used to collect data, including measurements of the long and short diameters of the tumors and animal weights. Raw data was measured using a balance and calipers and then directly imported into the software, and any changes to the data were recorded in the software. All processes, including drug administration, tumor measurement, and weight measurement, were performed in a biological safety cabinet or ultra-clean bench.

[0202] All experimental results were expressed as mean tumor volume ± SEM (standard error of the mean). The optimal drug treatment time point (usually after the last dose) was selected for statistical analysis between different groups. An independent sample t-test was used to compare relative tumor volume and tumor weight between treatment and control groups to determine whether there were significant differences. All data were analyzed using SPSS 18.0. p<0.05 was considered to indicate a significant difference.

[0203] The experimental results showed that the compound of the present invention administered at a dose of 5 mg / kg BID (twice daily) for 37 days showed significant tumor suppression effects in the in vivo efficacy experiment of HuPrime® gastric cancer GA3055 subcutaneous xenograft female BALB / C nude mice model, and the results are shown in Table 12 below.

[0204] [Table 12] Experimental conclusion: The compounds of the present invention had good tumor inhibitory effects in HuPrime® gastric cancer GA3055 subcutaneous xenograft female BALB / C nude mice model.

[0205] Experimental Example 4: Pharmacokinetic evaluation Test materials: CD-1 mice (male, 7-9 weeks old, Shanghai Slake) Experimental Procedure: The pharmacokinetic properties of the compounds in rodents after intravenous (IV) and oral administration (PO) were studied using standard protocols. Experiments were performed in mice with single intravenous injections and oral administration. The vehicle for intravenous injections was water. The oral vehicle was 0.5% hydroxypropylmethylcellulose. In this project, four male CD-1 mice were used. Two mice were intravenously injected with 0.5 mg / kg, and plasma samples were collected at time 0 (before administration) and 0.083, 0.25, 0.5, 1, 2, 4, 8, and 24 hours after administration. The other two mice were orally administered with 2.5 mg / kg intragastrically, and plasma samples were collected at time 0 (before administration) and 0.25, 0.5, 1, 2, 4, 8, and 24 hours after administration. Whole blood samples were collected within 24 hours, centrifuged at 3000 g for 15 minutes, and the supernatant was separated to obtain plasma samples. Four volumes of acetonitrile solution containing an internal standard were added to precipitate proteins, and the supernatant was centrifuged and collected. An equal volume of water was added, centrifuged, and the supernatant was collected for analysis. Blood drug concentrations were quantitatively analyzed by LC-MS / MS analysis, and the peak concentration (C max ), clearance (CL), half-life (T 1 / 2 ), tissue distribution (Vdss), area under the drug concentration-time curve (AUC 0-last Pharmacokinetic parameters such as bioavailability (F) were calculated.

[0206] The experimental results are shown in Table 13.

[0207] [Table 13] Experimental conclusion: The compounds of the present invention have good pharmacokinetic properties.

[0208] Experimental Example 5: Toxicity evaluation experiment in rats administered continuously for 21 days Test materials: SD rats (male and female, 7–8 weeks old, Zhejiang Vital River Laboratory Animal Technology Co., Ltd.).

[0209] Experimental Procedure: In this study, the potential toxicity of a candidate cancer treatment drug was detected by orally administering it to SD rats once daily for 21 days, while the toxicokinetic (TK) properties of the compound in animals were also investigated.

[0210] One hundred and twenty-six rats (63 per sex) were divided into six groups. The main test groups consisted of five males and five females. The TK test control groups consisted of three males and three females. The treatment groups consisted of six males and six females. The compounds were administered once daily at doses of 1, 3, 10, or 20 mg per kg per day (mg / kg / day). The control group received vehicle only. The dose volume was 10 mL / kg. At the start of treatment, the animals were 7–8 weeks old, with weights ranging from 251.11–308.11 g for males and 181.53–219.03 g for females.

[0211] Endpoints included mortality (moribund / dead), clinical signs, body weight, food intake, clinical pathology (hematology, serum biochemistry, blood coagulation), gross (necropsy) evaluation, organ weights, histopathological evaluation, and toxicokinetics. Toxicokinetic parameters included peak concentration (C max ), time to maximum drug concentration (T max ), area under the drug concentration-time curve (AUC 0-last ) were included.

[0212] The experimental results are shown in Table 14.

[0213] [Table 14] Experimental conclusion: The compounds of the present invention showed no significant decrease in animal body weight during administration and were well tolerated. Compared to the vehicle control group, fibrinogen and total bilirubin levels were mainly slightly increased, trabecular bone / cortical bone was decreased, and growth plate was thickened, but there were no other significant abnormalities. In toxicity evaluation of rats administered the compounds of the present invention for 21 consecutive days, they showed high maximum tolerated doses and high maximum tolerated exposure levels, indicating the safety of the compounds of the present invention.

Claims

1. A compound of formula (X) or a pharmaceutically acceptable salt thereof: 【Chemistry 1】 (however, T is selected from CR and N; T １ is selected from CH and N; T ２ is selected from CH and N; T ３ is selected from CH and N; L is -C(R ４ R ５ ) - and Ring A is 【Chemistry 2】 Each R １ are each independently F, Cl, Br, I, C １－３ Alkyl and C １－３ alkoxy; Each R ２ is H, Each R ３ is C １－３ is alkyl, R ４ and R ５ is H, R is H, F, Cl, Br, I, C １－３ Alkyl and C １－３ alkoxy; m is selected from 0, 1, 2 and 3; n is 1, p is selected from 0, 1, 2 and 3.

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, which satisfies the following (1) to (3): (1) Each R １ are each independently F, Cl, Br, I, or —CH ３ and -OCH ３ Selected from: (2) Each R ３ is -CH ３ and (3) R is H, F, Cl, Br, I, or —CH ３ is selected from.

3. The compound according to claim 2 or a pharmaceutically acceptable salt thereof, which satisfies the following (1) and (2): (1) Each R １ are each independently F, —CH ３ and -OCH ３ selected from: (2) R is selected from H and F.

4. The compound according to claim 1, wherein the compound has a structure represented by formula (X-4), (X-5), or (X-6), or a pharmaceutically acceptable salt thereof. 【Transformation 3】 (However, R １ , R ３ , T, T １ , T ２ , T ３ , m and p are as defined in claim 1.

5. The compound according to claim 4, wherein the compound has a structure represented by formula (X-7), (X-8), or (X-9), or a pharmaceutically acceptable salt thereof. 【Chemistry 4】 (However, R １ , R ３ , T, T １ , T ２ and T ３ is as defined in claim 4.

6. A compound of the formula: or a pharmaceutically acceptable salt thereof. 【Transformation 5】

7. A type A crystal of Compound 2, characterized in that the powder X-ray diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 7.45±0.20°, 13.53±0.20°, 13.94±0.20°, and 15.93±0.20°. 【Transformation 6】

8. The A-type crystal according to claim 7, characterized in that the powder X-ray diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 7.45±0.20°, 10.56±0.20°, 13.53±0.20°, 13.94±0.20°, 14.86±0.20°, 15.93±0.20°, and 17.96±0.20°.

9. The A-type crystal according to claim 8, characterized in that the powder X-ray diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 4.96±0.20°, 7.45±0.20°, 10.56±0.20°, 13.53±0.20°, 13.94±0.20°, 14.86±0.20°, 15.93±0.20°, 17.96±0.20°, 19.87±0.20°, 20.90±0.20°, 25.38±0.20°, and 28.02±0.20°.

10. The A-type crystal according to claim 9, characterized in that the powder X-ray diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 4.96±0.20°, 7.45±0.20°, 10.56±0.20°, 13.53±0.20°, 13.94±0.20°, 14.86±0.20°, 15.93±0.20°, 17.96±0.20°, 19.06±0.20°, 19.87±0.20°, 20.90±0.20°, 24.11±0.20°, 24.97±0.20°, 25.38±0.20°, and 28.02±0.20°.

11. The A-type crystal according to claim 10, characterized in that the powder X-ray diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 4.96°, 7.45°, 10.56°, 13.53°, 13.94°, 14.86°, 15.93°, 17.56°, 17.96°, 19.06°, 19.87°, 20.90°, 21.21°, 21.90°, 22.63°, 24.11°, 24.97°, 25.38°, 25.96°, 27.14°, 28.02°, 28.73°, 29.94°, 30.89°, 32.50°, 34.01°, 35.05°, 35.95°, 37.54°, and 39.08°.

12. The A-type crystal according to claim 7, Analytical data for XRPD patterns shown in the table below: Table 1 A type crystal having the formula:

13. A type A crystal according to claim 7, which satisfies the following (1) and (2): (1) A type crystal having a differential scanning calorimetry (DSC) curve with endothermic peak onsets at 259.7°C ± 5°C and 274.7°C ± 5°C; and (2) Thermogravimetric analysis (TGA) curve shows a weight loss of 0.80% at 240°C±3°C, and is an A-type crystal.

14. The compound according to any one of claims 1 to 6 or a pharmaceutically acceptable salt thereof, or the A-type crystal according to any one of claims 7 to 13, for use in inhibiting porcupine.

15. The compound according to any one of claims 1 to 6 or a pharmaceutically acceptable salt thereof, or the A-type crystal according to any one of claims 7 to 13, for use in the treatment of pancreatic cancer, colorectal cancer, and / or gastric cancer.

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