Salt and crystal form of nitrogen-containing heterocyclic derivative, preparation method therefor and application thereof

Abstract

A salt and a crystal form relating to a nitrogen-containing heterocyclic derivative, a preparation method therefor and an application thereof. In particular, the present invention relates to a salt and crystal form of a compound represented by general formula (I), a preparation method therefor, a pharmaceutical composition comprising a therapeutically effective amount of the crystal form, and a use thereof as a KRAS G12C mutation inhibitor in the treatment of diseases or conditions such as leukemia, neuroblastoma, melanoma, breast cancer, lung cancer and colon cancer. Each substituent in the general formula (I) is the same as defined in the description.

Description

Salts, crystal forms, preparation methods, and applications of nitrogen-containing heterocyclic derivatives

[0001] This application claims priority to Chinese Patent Application No. 2020113542899 filed on November 26, 2020, and Chinese Patent Application No. 2021113892168 filed on November 22, 2021. This application incorporates the entirety of the aforementioned Chinese patent applications. Technical Field

[0002] The present invention belongs to the field of biomedicine, and specifically relates to a salt, a crystal form, a preparation method and an application of a nitrogen-containing heterocyclic derivative. Background Art

[0003] Rat sarcoma (RAS) is encoded by the proto-oncogenes HRAS, NRAS, and KRAS, which are divided into four proteins: HRAS, NRAS, KRAS4A, and KRAS4B. RAS is a GTP (guanosine triphosphate)-binding protein located on the inner surface of the cell membrane. Its upstream is the receptor tyrosine kinase (RTK). Upon activation, RAS regulates downstream signaling pathways such as PI3K and RAF, thereby controlling cell growth, survival, migration, and differentiation.

[0004] RAS exists in two main states within the body: an inactive state bound to GDP (guanosine diphosphate) and an activated state bound to GTP. Its activity is regulated by two proteins: the guanine nucleotide exchange factor (GEF), which releases GDP from the RAS protein, allowing GTP to bind and activate RAS; and the GTPase activating protein (GAP), which activates the GTPase activity of the RAS protein, hydrolyzing the bound GTP to GDP, thereby inactivating RAS. Under normal circumstances, the RAS protein is in an inactive state. However, mutations alter its conformation, leaving RAS in a persistently activated state and continuously activating downstream signaling pathways, leading to the development of various cancers.

[0005] As the first oncogene to be identified, RAS is the most frequently mutated oncogene, accounting for an average of 25% of human cancers. The most common oncogenic mutation in the RAS family is KRAS (85%), while NRAS (12%) and HRAS (3%) are less common. KRAS mutations are prevalent in a range of cancers, including pancreatic cancer (95%), colorectal cancer (52%), and lung cancer (31%). The most common KRAS mutation is a point mutation, occurring at G12, G13, and Q61 of the Switch II region (aa59-76) within the p-loop (aa 10-17), with G12 mutation being the most common (83%). KRAS G12C is one of the most common mutations in non-small cell lung cancer (NSCLC) and colorectal cancer.

[0006] Despite the immense clinical need, no drug directly targeting KRAS has yet been marketed. Currently, chemotherapy is the only treatment for patients with KRAS mutations. The development of KRAS inhibitors is hampered by two main factors: first, the smooth structure of the RAS protein makes it difficult for small molecules to bind to the protein surface; second, the affinity of RAS GTPase for GTP is as high as the picomolar (pM) level, and endogenous GTP levels are high, making it difficult for small molecule drugs to block the binding of the two. Recent studies have found that when glycine (Gly) at position 12 of KRAS mutates to cysteine ​​(Cys), the conformation changes, forming a new pocket for covalent binding of small molecules, irreversibly locking KRAS G12C in an inactive state bound to GDP. Therefore, KRAS G12C inhibitors are expected to become the first drugs directly targeting KRAS.

[0007] Currently, several KRAS G12C inhibitors have entered the clinical research stage, such as AMG 510 developed by Amgen, ARS-3248 developed by Wellspring Biosciences, and MTRX849 developed by Mirati. All of them are currently in the Phase I clinical research stage, but no KRAS G12C inhibitor has been developed and marketed.

[0008] There is currently no specific targeted drug for KRAS G12C, and there is a large clinical need. KRAS G12C inhibitors with higher selectivity, better activity, and better safety have the potential to treat various cancers and have broad market prospects.

[0009] Jiangsu Hausen Pharmaceuticals Group Co., Ltd.'s patent application (application number: PCT / CN2020 / 093285) discloses the structures of a series of pyridazine derivative inhibitors. In subsequent research and development, in order to make the products easy to handle, filter, dry, and store, and to ensure long-term stability and high bioavailability, the present invention has conducted a comprehensive study of the salts and crystal forms of the above-mentioned substances, and is committed to obtaining the most suitable crystal form.

[0010] Summary of the Invention

[0011] All contents involved in patent application PCT / CN2020 / 093285 are added to the present invention by citation.

[0012] The object of the present invention is to provide an acid salt of a compound represented by general formula (I):

[0013]

[0014] in:

[0015] R a Each is independently selected from hydrogen, deuterium, halogen, amino, hydroxyl, mercapto, cyano, nitro, alkyl, deuterated alkyl, haloalkyl, alkoxy, -SR aa 、-C(O)R aa 、-NR aa R bb , haloalkoxy or hydroxyalkyl;

[0016] R1 is selected from hydrogen, deuterium, halogen, amino, hydroxy, cyano, nitro, alkyl, deuterated alkyl, haloalkyl, alkoxy, -SR aa 、-C(O)R aa 、-NR aa R bb , haloalkoxy or hydroxyalkyl;

[0017] R2 is selected from alkyl;

[0018] R3 is selected from hydrogen, deuterium, halogen, amino, hydroxy, cyano, nitro, alkyl, deuterated alkyl, haloalkyl, alkoxy, -SR aa 、-C(O)R aa 、-NR aa R bb or hydroxyalkyl;

[0019] R4 is selected from hydrogen, deuterium, halogen, amino, hydroxy, cyano, nitro, alkyl, deuterated alkyl, haloalkyl, alkoxy, -SR aa 、-C(O)R aa 、-NR aa R bb or hydroxyalkyl;

[0020] R5 is selected from hydrogen, deuterium, halogen, amino, hydroxy, mercapto, cyano, nitro, alkyl, deuterated alkyl, haloalkyl, alkoxy, -SR aa 、-C(O)R aa 、-NR aa R bb or hydroxyalkyl;

[0021] R6 is selected from hydrogen, deuterium, halogen, amino, hydroxy, mercapto, cyano, nitro, alkyl, deuterated alkyl, haloalkyl, alkoxy, -SR aa 、-C(O)R aa 、-NR aa R bb or hydroxyalkyl;

[0022] R7 is selected from hydrogen, deuterium, halogen, amino, hydroxy, mercapto, cyano, nitro, alkyl, deuterated alkyl, haloalkyl, alkoxy, -SR aa 、-C(O)R aa 、-NR aa R bb or hydroxyalkyl;

[0023] R aa is selected from deuterium, halogen, alkyl, deuterated alkyl or haloalkyl;

[0024] R bb is selected from deuterium, halogen, alkyl, deuterated alkyl, or haloalkyl; and

[0025] x is selected from 0, 1, 2 or 3.

[0026] In a preferred embodiment of the present invention, in the acid salt of the compound represented by general formula (I), R a Each independently selected from hydrogen, deuterium, halogen, amino, hydroxyl, thiol, cyano, nitro, C 1-6 Alkyl, C 1-6 Deuterated alkyl, C 1-6 Halogenated alkyl, C 1-6 Alkoxy, -SR aa 、-C(O)R aa 、-NR aa R bb 、C 1-6 Haloalkoxy or C 1-6 hydroxyalkyl;

[0027] Preferred are hydrogen, deuterium, halogen, amino, hydroxyl, mercapto, cyano, nitro, C 1-3 Alkyl, C 1-3 Deuterated alkyl, C 1- 3 haloalkyl, C 1-3 Alkoxy, -SR aa 、-C(O)R aa、-NR aa R bb 、C 1-3 Haloalkoxy or C 1-3 hydroxyalkyl;

[0028] More preferably, hydrogen, deuterium, halogen, amino, hydroxyl, mercapto, cyano, nitro, methyl, ethyl, propyl, isopropyl, deuterated methyl, deuterated ethyl, deuterated propyl, deuterated isopropyl, halomethyl, haloethyl, halopropyl, haloisopropyl, methoxy, ethoxy, propoxy, isopropoxy, methylthio, ethylthio, propylthio, isopropylthio, halomethoxy, haloethoxy, halopropoxy, hydroxymethyl, hydroxyethyl, hydroxypropyl or hydroxyisopropyl;

[0029] R1 is selected from hydrogen, deuterium, halogen, amino, hydroxy, cyano, nitro, C 1-6 Alkyl, C 1-6 Deuterated alkyl, C 1-6 Halogenated alkyl, C 1-6 Alkoxy, -SR aa 、-C(O)R aa 、-NR aa R bb 、C 1-6 Haloalkoxy or C 1-6 hydroxyalkyl;

[0030] Preferred are hydrogen, deuterium, halogen, amino, hydroxyl, cyano, nitro, C 1-3 Alkyl, C 1-3 Deuterated alkyl, C 1-3 Halogenated alkyl, C 1-3 Alkoxy, -SR aa 、-C(O)R aa 、-NR aa R bb 、C 1-3 Haloalkoxy or C 1-3 hydroxyalkyl;

[0031] More preferably hydrogen, methyl, fluorine, chlorine, amino, hydroxyl or cyano;

[0032] R2 is selected from hydrogen or C 1-6 alkyl;

[0033] Preferably hydrogen or C 1-3 alkyl;

[0034] More preferably hydrogen, methyl, ethyl, propyl or isopropyl;

[0035] R3 is selected from hydrogen, deuterium, halogen, amino, hydroxy, cyano, nitro, C 1-6 Alkyl, C 1-6 Deuterated alkyl, C 1-6 Halogenated alkyl, C1-6 Alkoxy, -SR aa 、-C(O)R aa 、-NR aa R bb or C 1-6 hydroxyalkyl;

[0036] Preferred are hydrogen, deuterium, halogen, amino, hydroxyl, cyano, nitro, C 1-3 Alkyl, C 1-3 Deuterated alkyl, C 1-3 Halogenated alkyl, C 1-3 Alkoxy, -SR aa or C 1-3 hydroxyalkyl;

[0037] More preferably, hydrogen, deuterium, fluorine, chlorine, bromine, iodine, amino, hydroxyl, cyano, nitro, methyl, ethyl, propyl, isopropyl, deuterated methyl, deuterated ethyl, deuterated propyl, deuterated isopropyl, halomethyl, haloethyl, halopropyl, haloisopropyl, methoxy, ethoxy, propoxy, isopropoxy, -S(CH 3 ), hydroxymethyl, hydroxyethyl, hydroxypropyl or hydroxyisopropyl;

[0038] R4 is selected from hydrogen, deuterium, halogen, amino, hydroxy, cyano, nitro, C 1-6 Alkyl, C 1-6 Deuterated alkyl, C 1-6 Halogenated alkyl, C 1-6 Alkoxy, -SR aa 、-C(O)R aa 、-NR aa R bb , or C 1-6 hydroxyalkyl;

[0039] Preferred are hydrogen, deuterium, halogen, amino, hydroxyl, cyano, nitro, C 1-3 Alkyl, C 1-3 Deuterated alkyl, C 1-3 Halogenated alkyl, C 1-3 Alkoxy, -NR aa R bb , or C 1-3 hydroxyalkyl;

[0040] More preferably, hydrogen, deuterium, fluorine, chlorine, bromine, iodine, amino, hydroxyl, cyano, nitro, methyl, ethyl, propyl, isopropyl, deuterated methyl, deuterated ethyl, deuterated propyl, deuterated isopropyl, halomethyl, haloethyl, halopropyl, haloisopropyl, methoxy, ethoxy, propoxy, isopropoxy, -NH(CH 3 ), -N(CH 3 ) 2 , hydroxymethyl, hydroxyethyl, hydroxypropyl or hydroxyisopropyl;

[0041] R5 is selected from hydrogen, deuterium, halogen, amino, hydroxyl, mercapto, cyano, nitro, C1-6 Alkyl, C 1-6 Deuterated alkyl, C 1-6 Halogenated alkyl, C 1-6 Alkoxy, -SR aa 、-C(O)R aa 、-NR aa R bb or C 1-6 hydroxyalkyl;

[0042] Preferred are hydrogen, deuterium, halogen, amino, hydroxyl, mercapto, cyano, nitro, C 1-3 Alkyl, C 1-3 Deuterated alkyl, C 1- 3 haloalkyl, C 1-3 Alkoxy or C 1-3 hydroxyalkyl;

[0043] More preferably, hydrogen, deuterium, fluorine, chlorine, bromine, iodine, amino, hydroxyl, mercapto, cyano, nitro, methyl, ethyl, propyl, isopropyl, deuterated methyl, deuterated ethyl, deuterated propyl, deuterated isopropyl, halomethyl, haloethyl, halopropyl, haloisopropyl, methoxy, ethoxy, propoxy, isopropoxy, hydroxymethyl, hydroxyethyl, hydroxypropyl or hydroxyisopropyl;

[0044] R6 is selected from hydrogen, deuterium, halogen, amino, hydroxyl, mercapto, cyano, nitro, C 1-6 Alkyl, C 1-6 Deuterated alkyl, C 1-6 Halogenated alkyl, C 1-6 Alkoxy, -SR aa 、-C(O)R aa 、-NR aa R bb or C 1-6 hydroxyalkyl;

[0045] Preferred are hydrogen, deuterium, halogen, amino, hydroxyl, mercapto, cyano, nitro, C 1-3 Alkyl, C 1-3 Deuterated alkyl, C 1- 3 haloalkyl, C 1-3 Alkoxy or C 1-3 hydroxyalkyl;

[0046] More preferably, hydrogen, deuterium, fluorine, chlorine, bromine, iodine, amino, hydroxyl, mercapto, cyano, nitro, methyl, ethyl, propyl, isopropyl, deuterated methyl, deuterated ethyl, deuterated propyl, deuterated isopropyl, halomethyl, haloethyl, halopropyl, haloisopropyl, methoxy, ethoxy, propoxy, isopropoxy, hydroxymethyl, hydroxyethyl, hydroxypropyl or hydroxyisopropyl;

[0047] R7 is selected from hydrogen, deuterium, halogen, amino, hydroxyl, mercapto, cyano, nitro, C1-6 Alkyl, C 1-6 Deuterated alkyl, C 1-6 Halogenated alkyl, C 1-6 Alkoxy, -SR aa 、-C(O)R aa 、-NR aa R bb or C 1-6 hydroxyalkyl;

[0048] Preferred are hydrogen, deuterium, halogen, amino, hydroxyl, mercapto, cyano, nitro, C 1-3 Alkyl, C 1-3 Deuterated alkyl, C 1- 3 haloalkyl, C 1-3 Alkoxy or C 1-3 hydroxyalkyl;

[0049] More preferably, hydrogen, deuterium, fluorine, chlorine, bromine, iodine, amino, hydroxyl, mercapto, cyano, nitro, methyl, ethyl, propyl, isopropyl, deuterated methyl, deuterated ethyl, deuterated propyl, deuterated isopropyl, halomethyl, haloethyl, halopropyl, haloisopropyl, methoxy, ethoxy, propoxy, isopropoxy, hydroxymethyl, hydroxyethyl, hydroxypropyl or hydroxyisopropyl;

[0050] R aa Selected from deuterium, halogen, C 1-6 Alkyl, C 1-6 Deuterated alkyl or C 1-6 alkyl halide;

[0051] Deuterium, halogen, C 1-3 Alkyl, C 1-3 Deuterated alkyl or C 1-3 alkyl halide;

[0052] More preferably methyl, ethyl, propyl or isopropyl;

[0053] R bb Selected from deuterium, halogen, C 1-6 Alkyl, C 1-6 Deuterated alkyl or C 1-6 alkyl halide;

[0054] Deuterium, halogen, C 1-3 Alkyl, C 1-3 Deuterated alkyl or C 1-3 alkyl halide;

[0055] More preferably methyl, ethyl, propyl or isopropyl;

[0056] x is selected from 0, 1, 2 or 3; preferably 0, 1 or 2; more preferably 0 or 1.

[0057] In a preferred embodiment of the present invention, the acid salt of the compound is represented by the general formula (II):

[0058]

[0059] in:

[0060] R a is selected from hydrogen or methyl;

[0061] R1 is selected from hydrogen, fluorine, chlorine, bromine or methyl;

[0062] R3 is selected from hydrogen, amino, hydroxy, fluoro, chloro, methyl, -S(CH3) or trifluoromethyl;

[0063] R4 is selected from hydrogen, amino, hydroxy, fluorine, chlorine, -N(CH3)2, -NH(CH3) or fluorine;

[0064] R5 is selected from hydrogen, fluorine, chlorine, bromine, methyl, ethyl, propyl or isopropyl;

[0065] R6 is selected from hydrogen, fluorine, chlorine, bromine, methyl, ethyl, propyl or isopropyl;

[0066] R7 is selected from hydrogen, fluorine, chlorine, bromine or methyl.

[0067] In a preferred embodiment of the present invention, the acid salt of the compound is further represented by the general formula (II-A) or (II-B):

[0068]

[0069] In a preferred embodiment of the present invention, the acid salt of the compound, wherein the compound is selected from:

[0070]

[0071]

[0072]

[0073] In a more preferred embodiment of the present invention, the acid salt of the compound, wherein the compound is selected from:

[0074]

[0075]

[0076] The acid in the acid salt is selected from isethionic acid, sulfuric acid, 1,5-naphthalene disulfonic acid, methanesulfonic acid, hydrobromic acid, phosphoric acid, benzenesulfonic acid, oxalic acid, maleic acid, adipic acid, hydrochloric acid, citric acid, malonic acid, L-malic acid, pamoic acid, p-toluenesulfonic acid or fumaric acid, preferably isethionic acid or sulfuric acid.

[0077] In a further preferred embodiment of the present invention, the number of acids is 0.2-3; preferably 0.2, 0.5, 1, 1.5, 2, 2.5 or 3; more preferably 0.5, 1, 2 or 3.

[0078] In a further preferred embodiment of the present invention, the acid salt is a hydrate or an anhydrate, and when the acid salt is a hydrate, the number of water is 0.2-3; preferably 0.2, 0.5, 1, 1.5, 2, 2.5 or 3; more preferably 0.5, 1, 2 or 3.

[0079] In the most preferred embodiment of the present invention, an acid salt of the compound P-4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-7-(6-amino-3-chloro-2-fluorophenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one is provided, wherein the acid in the acid salt is selected from isethionic acid, sulfuric acid, 1,5-naphthalenedisulfonic acid, methanesulfonic acid, hydrobromic acid, phosphoric acid, benzenesulfonic acid, oxalic acid, maleic acid, adipic acid, hydrochloric acid, citric acid, malonic acid, L-malic acid, pamoic acid, p-toluenesulfonic acid or fumaric acid, wherein the structure of the acid salt of the compound is as follows:

[0080]

[0081]

[0082] In a preferred embodiment of the present invention, the acid salt is a crystalline form; preferably, the acid salt of the compound P-4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-7-(6-amino-3-chloro-2-fluorophenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one is a crystalline form;

[0083] A crystalline acid salt of P-4-((S)-4-acryloyl-2-methylpiperazin-1-yl)-6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one;

[0084] A crystalline acid salt of P-4-((S)-4-acryloyl-2-methylpiperazin-1-yl)-7-(2-amino-6-fluorophenyl)-6-fluoro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one;

[0085] A crystalline acid salt of P-4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-6-chloro-7-(2-fluoro-6-hydroxyphenyl)-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one;

[0086] A crystalline acid salt of P-4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one;

[0087] More preferred are isethionate salt crystalline form, sulfate salt crystalline form, 1,5-naphthalene disulfonate salt crystalline form, methanesulfonate salt crystalline form, hydrobromide salt crystalline form, phosphate salt crystalline form, benzenesulfonate salt crystalline form, oxalate salt crystalline form, maleate salt crystalline form, adipate salt crystalline form, hydrochloride salt crystalline form, citrate salt crystalline form, malonate salt crystalline form, L-malate salt crystalline form, pamoate salt crystalline form, p-toluenesulfonate salt crystalline form or fumarate salt crystalline form.

[0088] In a preferred embodiment of the present invention, a crystalline form of the acid salt of compound P-4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-7-(6-amino-3-chloro-2-fluorophenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one is provided.

[0089] In a more preferred embodiment of the present invention, the acid salt of compound P-4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-7-(6-amino-3-chloro-2-fluorophenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one is a crystalline form, preferably a hydroxyethyl sulfonate crystalline form, a sulfate crystalline form, a 1,5-naphthalene disulfonate crystalline form, a methanesulfonate crystalline form, a hydrobromide crystalline form, a phosphate crystalline form, a benzenesulfonate crystalline form, an oxalate crystalline form, a maleate crystalline form, an adipate crystalline form, a hydrochloride crystalline form, a citrate crystalline form, a malonate crystalline form, an L-malate crystalline form, a pamoate crystalline form, a p-toluenesulfonate crystalline form or a fumarate crystalline form.

[0090] In a preferred embodiment of the present invention, the acid salt is in crystalline form, wherein the number of acids is 0.2-3; preferably 0.2, 0.5, 1, 1.5, 2, 2.5 or 3; more preferably 0.5, 1, 2 or 3.

[0091] In a preferred embodiment of the present invention, the isethionate salt crystalline form I-III and the sulfate salt crystalline form I-IV of the compound P-4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-7-(6-amino-3-chloro-2-fluorophenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one are provided:

[0092] The X-ray powder diffraction pattern of the isethionate salt form I has a diffraction peak at 21.7±0.2°; or a diffraction peak at 8.8±0.2°; or a diffraction peak at 19.3±0.2°; or a diffraction peak at 27.6±0.2°; or a diffraction peak at 10.9±0.2°; or a diffraction peak at 15.4±0.2°; or a diffraction peak at 16.7±0.2°; or a diffraction peak at 15.8 ± 0.2 ° having a diffraction peak; or having a diffraction peak at 17.5 ± 0.2 °; or having a diffraction peak at 23.8 ± 0.2 °; or having a diffraction peak at 10.2 ± 0.2 °; or having a diffraction peak at 11.8 ± 0.2 °; preferably including any 2-5, or 3-5, or 3-6, or 3-8, or 5-8, or 6-8 of the above diffraction peaks, more preferably including any 6, 7 or 8 thereof;

[0093] The X-ray powder diffraction pattern of the isethionate salt form II has a diffraction peak at 21.7±0.2°; or a diffraction peak at 8.8±0.2°; or a diffraction peak at 19.3±0.2°; or a diffraction peak at 27.6±0.2°; or a diffraction peak at 10.9±0.2°; or a diffraction peak at 23.8±0.2°; or a diffraction peak at 16.7±0.2°; or a diffraction peak at 15.4±0.2°; or a diffraction peak at 15.8±0.2°; or a diffraction peak at 10.0±0.2°; preferably includes any 2-5, or 3-5, or 3-6, or 3-8, or 5-8, or 6-8 of the above diffraction peaks, and more preferably includes any 6, 7 or 8 thereof;

[0094] The X-ray powder diffraction pattern of the isethionate salt form III has a diffraction peak at 19.4±0.2°; or a diffraction peak at 16.9±0.2°; or a diffraction peak at 26.6±0.2°; or a diffraction peak at 14.6±0.2°; or a diffraction peak at 28.0±0.2°; or a diffraction peak at 25.6±0.2°; or a diffraction peak at 20.7±0.2°; or a diffraction peak at 12.8±0.2°; or a diffraction peak at 19.1±0.2°; or a diffraction peak at 27.2±0.2°; preferably includes any 2-5, or 3-5, or 3-6, or 3-8, or 5-8, or 6-8 of the above diffraction peaks, and more preferably includes any 6, 7 or 8 thereof;

[0095] The X-ray powder diffraction pattern 2θ of the sulfate salt crystalline form I has a diffraction peak at 19.0±0.2°; or a diffraction peak at 19.4±0.2°; or a diffraction peak at 12.4±0.2°; or a diffraction peak at 26.2±0.2°; or a diffraction peak at 17.6±0.2°; or a diffraction peak at 18.1±0.2°; or a diffraction peak at 25.3±0.2°; or a diffraction peak at 8.8±0.2°; or a diffraction peak at 21.9±0.2°; or a diffraction peak at 11.5±0.2°; preferably comprising any 2-5, or 3-5, or 3-6, or 3-8, or 5-8, or 6-8 of the above diffraction peaks, more preferably comprising any 6, 7 or 8 thereof;

[0096] The X-ray powder diffraction pattern 2θ of the sulfate salt crystalline form II has a diffraction peak at 15.5±0.2°; or a diffraction peak at 11.1±0.2°; or a diffraction peak at 8.9±0.2°; or a diffraction peak at 19.3±0.2°; or a diffraction peak at 22.3±0.2°; or a diffraction peak at 23.6±0.2°; or a diffraction peak at 17.4±0.2°; or a diffraction peak at 27.3±0.2°; or a diffraction peak at 17.0±0.2°; or a diffraction peak at 27.9±0.2°; preferably comprising any 2-5, or 3-5, or 3-6, or 3-8, or 5-8, or 6-8 of the above diffraction peaks, more preferably comprising any 6, 7 or 8 thereof;

[0097] The X-ray powder diffraction pattern 2θ of the sulfate salt crystalline form III has a diffraction peak at 19.6±0.2°; or a diffraction peak at 18.0±0.2°; or a diffraction peak at 18.4±0.2°; or a diffraction peak at 16.8±0.2°; or a diffraction peak at 14.3±0.2°; or a diffraction peak at 11.8±0.2°; or a diffraction peak at 14.9±0.2°; or a diffraction peak at 25.7±0.2°; or a diffraction peak at 15.4±0.2°; or a diffraction peak at 23.5±0.2°; preferably comprising any 2-5 of the above diffraction peaks, or 3-5, or 3-6, or 3-8, or 5-8, or 6-8, more preferably comprising any 6, 7 or 8 thereof;

[0098] The X-ray powder diffraction pattern 2θ of sulfate form IV has a diffraction peak at 19.4±0.2°; or a diffraction peak at 18.9±0.2°; or a diffraction peak at 15.5±0.2°; or a diffraction peak at 8.8±0.2°; or a diffraction peak at 18.1±0.2°; or a diffraction peak at 24.9±0.2°; or a diffraction peak at 17.4±0.2°; or a diffraction peak at 12.3±0.2°; or a diffraction peak at 26.1±0.2°; or a diffraction peak at 14.5±0.2°; preferably includes any 2-5 of the above diffraction peaks, or 3-5, or 3-6, or 3-8, or 5-8, or 6-8, more preferably includes any 6, 7 or 8 thereof.

[0099] In a further preferred embodiment of the present invention, the isethionate salt crystalline form I-III and the sulfate salt crystalline form I-IV of the compound P-4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-7-(6-amino-3-chloro-2-fluorophenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one are provided:

[0100] The X-ray powder diffraction pattern of the isethionate salt crystalline form I comprises at least one or more diffraction peaks located at 2θ of 21.7±0.2°, 8.8±0.2°, and 19.3±0.2°, preferably two of them, and more preferably three of them; optionally, it may further comprise at least one diffraction peak located at 2θ of 27.6±0.2°, 10.9±0.2°, 15.4±0.2°, 16.7±0.2°, 15.8±0.2°, 10.2±0.2°, and 11.8±0.2°, preferably two, three, four, or five of them; for example,

[0101] 21.7±0.2°, 8.8±0.2°;

[0102] 8.8±0.2°, 27.6±0.2°;

[0103] 21.7±0.2°, 8.8±0.2°, 10.9±0.2°;

[0104] 8.8±0.2°, 19.3±0.2°, 15.4±0.2°;

[0105] 21.7±0.2°, 8.8±0.2°, 27.6±0.2°, 10.9±0.2°;

[0106] 8.8±0.2°, 19.3±0.2°, 15.4±0.2°, 16.7±0.2°;

[0107] 15.8±0.2°, 8.8±0.2°, 27.6±0.2°, 10.9±0.2°;

[0108] 11.7±0.2°, 8.8±0.2°, 27.6±0.2°, 10.9±0.2°;

[0109] 16.7±0.2°, 8.8±0.2°, 19.3±0.2°, 16.7±0.2°, 10.9±0.2°, 15.4±0.2°;

[0110] 21.7±0.2°, 8.8±0.2°, 19.3±0.2°, 15.8±0.2°, 10.9±0.2°, 15.4±0.2°;

[0111] 10.9±0.2°, 8.8±0.2°, 10.2±0.2°, 27.6±0.2°, 10.9±0.2°, 15.8±0.2°;

[0112] The X-ray powder diffraction pattern of the isethionate salt form II comprises at least one or more diffraction peaks located at 2θ of 21.7±0.2°, 10.0±0.2°, and 8.8±0.2°, preferably two of them, and more preferably three of them; optionally, it may further comprise at least one diffraction peak located at 2θ of 19.3±0.2°, 27.6±0.2°, 10.9±0.2°, 23.8±0.2°, and 16.7±0.2°, preferably two, three, four, or five of them; for example,

[0113] 21.7±0.2°, 10.0±0.2°;

[0114] 10.0±0.2°, 8.8±0.2°;

[0115] 21.7±0.2°, 10.0±0.2°, 19.3±0.2°;

[0116] 10.0±0.2°, 8.8±0.2°, 27.6±0.2°;

[0117] 21.7±0.2°, 10.0±0.2°, 8.8±0.2°, 19.3±0.2°;

[0118] 10.0±0.2°, 8.8±0.2°, 19.3±0.2°, 27.6±0.2°;

[0119] 27.6±0.2°, 10.0±0.2°, 8.8±0.2°, 19.3±0.2°, 16.7±0.2°, 10.9±0.2°;

[0120] 21.7±0.2°, 10.0±0.2°, 8.8±0.2°, 16.7±0.2°, 27.6±0.2°, 10.9±0.2°;

[0121] The X-ray powder diffraction pattern of the isethionate salt form III comprises at least one or more diffraction peaks located at 2θ of 19.4±0.2°, 16.9±0.2°, and 26.6±0.2°, preferably two of them, and more preferably three of them; optionally, it may further comprise at least one diffraction peak located at 2θ of 14.6±0.2°, 28.0±0.2°, 25.6±0.2°, 20.7±0.2°, and 12.8±0.2°, preferably two, three, four, or five of them; for example,

[0122] 19.4±0.2°, 16.9±0.2°, 26.6±0.2°, 14.6±0.2°, 28.0±0.2°, 25.6±0.2°;

[0123] 19.4±0.2°, 16.9±0.2°, 26.6±0.2°, 12.8±0.2°, 28.0±0.2°, 25.6±0.2°;

[0124] The X-ray powder diffraction pattern of the sulfate salt crystalline form I comprises at least one or more diffraction peaks located at 2θ of 19.0±0.2°, 19.4±0.2°, and 12.4±0.2°, preferably two of them, and more preferably three of them; optionally, it may further comprise at least one of 2θ of 26.2±0.2°, 17.6±0.2°, 18.1±0.2°, 25.3±0.2°, and 8.8±0.2°, preferably two, three, four, or five of them; for example,

[0125] 19.0±0.2°, 19.4±0.2°, 12.4±0.2°, 26.2±0.2°, 17.6±0.2°, 18.1±0.2°;

[0126] 19.0±0.2°, 19.4±0.2°, 12.4±0.2°, 26.2±0.2°, 17.6±0.2°, 25.3±0.2°;

[0127] The X-ray powder diffraction pattern of the sulfate salt crystalline form II comprises at least one or more diffraction peaks located at 2θ of 15.5±0.2°, 11.1±0.2°, and 8.9±0.2°, preferably two of them, more preferably three; optionally, it may further comprise at least one of 2θ of 19.3±0.2°, 22.3±0.2°, 23.6±0.2°, 17.4±0.2°, and 27.3±0.2°, preferably 2, 3, 4 or 5 of them; for example,

[0128] 15.5±0.2°, 11.1±0.2°;

[0129] 11.1±0.2°, 8.9±0.2°;

[0130] 15.5±0.2°, 11.1±0.2°, 8.9±0.2°;

[0131] 11.1±0.2°, 8.9±0.2°, 19.3±0.2°;

[0132] 15.5±0.2°, 11.1±0.2°, 8.9±0.2°, 19.3±0.2°;

[0133] 15.5±0.2°, 11.1±0.2°, 8.9±0.2°, 19.3±0.2°, 22.3±0.2°, 27.3±0.2°;

[0134] 8.9±0.2°, 19.3±0.2°, 22.3±0.2°, 23.6±0.2°, 17.4±0.2°, 27.3±0.2°;

[0135] The X-ray powder diffraction pattern of the sulfate salt form III comprises at least one or more diffraction peaks located at 2θ of 19.6±0.2°, 18.0±0.2°, and 18.4±0.2°, preferably two of them, more preferably three; optionally, it may further comprise at least one of 2θ of 16.8±0.2°, 14.3±0.2°, 11.8±0.2°, 14.9±0.2°, and 25.7±0.2°, preferably 2, 3, 4 or 5 of them; for example,

[0136] 19.6±0.2°, 18.0±0.2°, 18.4±0.2°, 16.8±0.2°, 14.3±0.2°, 11.8±0.2°;

[0137] 19.6±0.2°, 18.0±0.2°, 18.4±0.2°, 16.8±0.2°, 14.3±0.2°, 14.9±0.2°;

[0138] The X-ray powder diffraction pattern of the sulfate salt form IV comprises at least one or more diffraction peaks located at 2θ of 19.4±0.2°, 18.9±0.2°, and 15.5±0.2°, preferably two of them, more preferably three; optionally, it may further comprise at least one of 2θ of 8.8±0.2°, 18.1±0.2°, 24.9±0.2°, 17.4±0.2°, and 12.3±0.2°, preferably 2, 3, 4 or 5 of them; for example,

[0139] 19.4±0.2°, 18.9±0.2°, 15.5±0.2°, 8.8±0.2°, 18.1±0.2°, 24.9±0.2°;

[0140] 19.4±0.2°, 18.9±0.2°, 15.5±0.2°, 8.8±0.2°, 12.3±0.2°, 24.9±0.2°.

[0141] In a further preferred embodiment of the present invention, the isethionate crystalline form I-III and the sulfate crystalline form I-IV of the compound P-4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-7-(6-amino-3-chloro-2-fluorophenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one are provided:

[0142] The X-ray powder diffraction pattern of the isethionate salt form I optionally further comprises one or more diffraction peaks located at 2θ of 21.7±0.2°, 8.8±0.2°, 10.2±0.2°, 11.8±0.2°, 13.3±0.2°, 19.3±0.2°, 27.6±0.2°, 10.9±0.2°, 15.4±0.2°, and 16.7±0.2°; preferably, at least any 2-3, or 4-5, or 6-8 of these peaks are included; further preferably, any 2, 3, 4, 5, 6, 7, or 8 of these peaks are included; for example,

[0143] 8.8±0.2°, 10.2±0.2°, 11.8±0.2°, 13.3±0.2°, 27.6±0.2°, 10.9±0.2°, 15.8±0.2°, 17.5±0.2°;

[0144] 27.6±0.2°, 10.9±0.2°, 15.4±0.2°, 17.5±0.2°, 15.8±0.2°, 16.7±0.2°, 17.5±0.2°, 23.8±0.2°;

[0145] 21.7±0.2°, 8.8±0.2°, 19.3±0.2°, 27.6±0.2°, 10.9±0.2°, 17.5±0.2°, 16.7±0.2°, 15.8±0.2°;

[0146] The X-ray powder diffraction pattern of the isethionate salt form II optionally further comprises one or more diffraction peaks located at 2θ of 10.0±0.2°, 21.7±0.2°, 8.8±0.2°, 19.3±0.2°, 27.6±0.2°, 10.9±0.2°, and 23.8±0.2°; preferably, at least any 2-3, or 4-5, or 6-8 of these peaks are included; further preferably, any 2, 3, 4, 5, 6, 7, or 8 of these peaks are included; for example,

[0147] 21.7 ± 0.2°, 8.8 ± 0.2°, 19.3 ± 0.2°, 27.6 ± 0.2°, 10.9 + ± 0.2°, 23.8 ± 0.2°, 16.7 ± 0.2°, 15.4 ± 0.2°;

[0148] 8.8 ± 0.2°, 19.3 ± 0.2°, 27.6 ± 0.2°, 10.9 + ± 0.2°, 23.8 ± 0.2°, 15.4 ± 0.2°, 15.8 ± 0.2°, 10.0 ± 0.2°;

[0149] The X-ray powder diffraction pattern of the isethionate salt form III optionally further comprises one or more diffraction peaks located at 2θ of 19.4±0.2°, 16.9±0.2°, 26.6±0.2°, 14.6±0.2°, 28.0±0.2°, 25.6±0.2°, and 20.7±0.2°; preferably, at least any 2-3, or 4-5, or 6-8 of these peaks are included; further preferably, any 2, 3, 4, 5, 6, 7, or 8 of these peaks are included; for example,

[0150] 19.4±0.2°, 16.9±0.2°, 26.6±0.2°, 14.6±0.2°, 28.0±0.2°, 25.6±0.2°, 20.7±0.2°, 27.2±0.2°;

[0151] 19.4±0.2°, 16.9±0.2°, 26.6±0.2°, 14.6±0.2°, 28.0±0.2°, 27.2±0.2°, 20.7±0.2°, 12.8±0.2°;

[0152] The X-ray powder diffraction pattern of the sulfate salt crystalline form I optionally further includes one or more diffraction peaks located at 2θ of 19.0±0.2°, 19.4±0.2°, 12.4±0.2°, 26.2±0.2°, 17.6±0.2°, 18.1±0.2°, and 25.3±0.2°; preferably, at least any 2-3, or 4-5, or 6-8 thereof; further preferably, any 2, 3, 4, 5, 6, 7, or 8 thereof; for example,

[0153] 19.0±0.2°, 19.4±0.2°, 12.4±0.2°, 26.2±0.2°, 17.6±0.2°, 18.1±0.2°, 25.3±0.2°, 8.8±0.2°;

[0154] The X-ray powder diffraction pattern of the sulfate salt form II optionally further comprises one or more diffraction peaks located at 2θ of 15.5±0.2°, 11.1±0.2°, 8.9±0.2°, 19.3±0.2°, 22.3±0.2°, 23.6±0.2°, and 17.4±0.2°; preferably, at least any 2-3, or 4-5, or 6-8 thereof; further preferably, any 2, 3, 4, 5, 6, 7, or 8 thereof; for example,

[0155] 15.5±0.2°, 8.9±0.2°, 19.3±0.2°, 22.3±0.2°, 23.6±0.2°, 17.4±0.2°, 27.3±0.2°, 17.0±0.2°;

[0156] 11.1±0.2°, 8.9±0.2°, 19.3±0.2°, 22.3±0.2°, 17.4±0.2°, 27.3±0.2°, 17.0±0.2°, 27.9±0.2°;

[0157] The X-ray powder diffraction pattern of the sulfate salt form III optionally further includes one or more diffraction peaks located at 2θ of 19.6±0.2°, 18.0±0.2°, 18.4±0.2°, 16.8±0.2°, 14.3±0.2°, 11.8±0.2°, and 14.9±0.2°; preferably, at least any 2-3, or 4-5, or 6-8 thereof; further preferably, any 2, 3, 4, 5, 6, 7, or 8 thereof; for example,

[0158] 19.6±0.2°, 18.0±0.2°, 18.4±0.2°, 16.8±0.2°, 14.3±0.2°, 11.8±0.2°, 14.9±0.2°, 15.4±0.2°;

[0159] 19.6±0.2°, 18.0±0.2°, 18.4±0.2°, 16.8±0.2°, 14.3±0.2°, 11.8±0.2°, 14.9±0.2°, 23.5±0.2°;

[0160] The X-ray powder diffraction pattern of the sulfate salt form IV optionally further comprises one or more diffraction peaks located at 2θ of 19.4±0.2°, 18.9±0.2°, 15.5±0.2°, 8.8±0.2°, 18.1±0.2°, 24.9±0.2°, and 17.4±0.2°; preferably, at least any 2-3, or 4-5, or 6-8 thereof; further preferably, any 2, 3, 4, 5, 6, 7, or 8 thereof; for example,

[0161] 19.4±0.2°, 18.9±0.2°, 15.5±0.2°, 8.8±0.2°, 18.1±0.2°, 24.9±0.2°, 17.4±0.2°, 12.3±0.2°;

[0162] 19.4±0.2°, 18.9±0.2°, 15.5±0.2°, 8.8±0.2°, 18.1±0.2°, 24.9±0.2°, 17.4±0.2°, 26.1±0.2°;

[0163] In a preferred embodiment of the present invention, the isethionate salt crystalline form I-III and the sulfate salt crystalline form I-IV of the compound P-4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-7-(6-amino-3-chloro-2-fluorophenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one are provided:

[0164] The X-ray powder diffraction pattern of the isethionate salt form I comprises 2θ values ​​of 21.7±0.2°, 8.8±0.2°, 10.2±0.2°, 11.8±0.2°, 13.1±0.2°, 19.3±0.2°, 27.6±0.2°, 10.9±0.2°, 13.3±0.2°, 15.4±0.2°, 16.7±0.2°. , 15.8±0.2°, 17.5±0.2°, 23.8±0.2°, 14.7±0.2°, 24.3±0.2°, 27.3±0.2°, 23.4±0.2°, 20.6±0.2°, 21.2±0.2°, one or more diffraction peaks, preferably, comprising 4, 5, 6, 8 or 10 diffraction peaks selected therefrom; for example,

[0165] 8.8±0.2°, 19.3±0.2°, 27.6±0.2°, 20.6±0.2°;

[0166] 19.3±0.2°, 10.9±0.2°, 15.4±0.2°, 16.7±0.2°;

[0167] 8.8±0.2°, 19.3±0.2°, 27.6±0.2°, 10.9±0.2°, 15.4±0.2°, 20.6±0.2°;

[0168] 19.3±0.2°, 27.6±0.2°, 10.9±0.2°, 16.7±0.2°, 23.4±0.2°, 20.6±0.2°;

[0169] 8.8±0.2°, 19.3±0.2°, 27.6±0.2°, 16.7±0.2°, 15.8±0.2°, 17.5±0.2°, 24.3±0.2°, 14.7±0.2°, 27.3±0.2°, 23.4±0.2°;

[0170] 21.7±0.2°, 8.8±0.2°, 19.3±0.2°, 27.6±0.2°, 10.9±0.2°, 15.4±0.2°, 16.7±0.2°, 15.8±0.2°, 24.3±0.2°, 23.8±0.2°;

[0171] 8.8±0.2°, 10.2±0.2°, 11.8±0.2°, 13.1±0.2°, 27.6±0.2°, 10.9±0.2°, 13.3±0.2°, 21.2±0.2°, 15.8±0.2°, 17.5±0.2°;

[0172] The X-ray powder diffraction pattern of the isethionate salt form II comprises one or more diffraction peaks located at 2θ of 21.7±0.2°, 10.0±0.2°, 8.8±0.2°, 19.3±0.2°, 27.6±0.2°, 10.9±0.2°, 23.8±0.2°, 16.7±0.2°, 15.4±0.2°, 15.8±0.2°, 17.5±0.2°, 14.7±0.2°, 24.4±0.2°, 27.3±0.2°, and 29.2±0.2°, preferably, comprises diffraction peaks at any of 4, 5, 6, 8, or 10 of the diffraction peaks; for example,

[0173] 10.0±0.2°, 8.8±0.2°, 19.3±0.2°, 29.2±0.2°;

[0174] 8.8±0.2°, 27.6±0.2°, 27.3±0.2°, 29.2±0.2°;

[0175] 21.7±0.2°, 10.0±0.2°, 8.8±0.2°, 19.3±0.2°, 27.6±0.2°, 29.2±0.2°;

[0176] 10.0±0.2°, 8.8±0.2°, 19.3±0.2°, 27.6±0.2°, 27.3±0.2°, 14.7±0.2°;

[0177] 21.7±0.2°, 8.8±0.2°, 19.3±0.2°, 27.6±0.2°, 10.9±0.2°, 23.8±0.2°, 27.3±0.2°, 17.5±0.2°;

[0178] 19.3±0.2°, 27.6±0.2°, 10.9±0.2°, 23.8±0.2°, 15.8±0.2°, , 16.7±0.2°, 15.4±0.2°, 17.5±0.2°;

[0179] 10.0±0.2°, 8.8±0.2°, 19.3±0.2°, 27.6±0.2°, 10.9±0.2°, 23.8±0.2°, 16.7±0.2°, 15.4±0.2°, 15.8±0.2°, 17.5±0.2°;

[0180] 8.8±0.2°, 19.3±0.2°, 27.6±0.2°, 10.9±0.2°, 23.8±0.2°, 16.7±0.2°, 15.4±0.2°, 17.5±0.2°, 27.3±0.2°, 29.2±0.2°;

[0181] The X-ray powder diffraction pattern of the isethionate salt form III comprises one or more diffraction peaks located at 2θ of 19.4±0.2°, 16.9±0.2°, 26.6±0.2°, 14.6±0.2°, 28.0±0.2°, 25.6±0.2°, 20.7±0.2°, 12.8±0.2°, 19.1±0.2°, 27.2±0.2°, 24.4±0.2°, 15.3±0.2°, 26.2±0.2°, 30.2±0.2°, and 27.4±0.2°, preferably, comprising 4, 5, 6, 8 or 10 diffraction peaks selected from these; for example,

[0182] 19.4±0.2°, 16.9±0.2°, 26.6±0.2°, 14.6±0.2°;

[0183] 19.4±0.2°, 16.9±0.2°, 26.6±0.2°, 28.0±0.2°;

[0184] 19.4±0.2°, 16.9±0.2°, 26.6±0.2°, 14.6±0.2°, 28.0±0.2°, 27.4±0.2°;

[0185] 19.4±0.2°, 16.9±0.2°, 26.6±0.2°, 14.6±0.2°, 28.0±0.2°, 30.2±0.2°;

[0186] 19.4±0.2°, 16.9±0.2°, 26.6±0.2°, 14.6±0.2°, 28.0±0.2°, 25.6±0.2°, 20.7±0.2°, 27.4±0.2°;

[0187] 19.4±0.2°, 16.9±0.2°, 26.6±0.2°, 14.6±0.2°, 28.0±0.2°, 25.6±0.2°, 20.7±0.2°, 30.2±0.2°;

[0188] 19.4±0.2°, 16.9±0.2°, 26.6±0.2°, 14.6±0.2°, 28.0±0.2°, 25.6±0.2°, 20.7±0.2°, 12.8±0.2°, 19.1±0.2°, 27.2±0.2°;

[0189] 19.4±0.2°, 16.9±0.2°, 26.6±0.2°, 14.6±0.2°, 28.0±0.2°, 25.6±0.2°, 20.7±0.2°, 12.8±0.2°, 19.1±0.2°, 24.4±0.2°;

[0190] The X-ray powder diffraction pattern of the sulfate salt crystalline form I comprises one or more diffraction peaks located at 2θ of 19.0±0.2°, 19.4±0.2°, 12.4±0.2°, 26.2±0.2°, 17.6±0.2°, 18.1±0.2°, 25.3±0.2°, 8.8±0.2°, 21.9±0.2°, and 11.5±0.2°, preferably, comprising 4, 5, 6, 8 or 10 diffraction peaks selected therefrom; for example,

[0191] 19.0±0.2°, 19.4±0.2°, 12.4±0.2°, 26.2±0.2°;

[0192] 19.0±0.2°, 19.4±0.2°, 12.4±0.2°, 17.6±0.2°;

[0193] 19.0±0.2°, 19.4±0.2°, 12.4±0.2°, 26.2±0.2°, 17.6±0.2°, 11.5±0.2°;

[0194] 19.0±0.2°, 19.4±0.2°, 12.4±0.2°, 26.2±0.2°, 17.6±0.2°, 21.9±0.2°;

[0195] 19.0±0.2°, 19.4±0.2°, 12.4±0.2°, 26.2±0.2°, 17.6±0.2°, 18.1±0.2°, 25.3±0.2°, 8.8±0.2°;

[0196] 19.0±0.2°, 19.4±0.2°, 12.4±0.2°, 26.2±0.2°, 17.6±0.2°, 18.1±0.2°, 25.3±0.2°, 21.9±0.2°;

[0197] 19.0±0.2°, 19.4±0.2°, 12.4±0.2°, 26.2±0.2°, 17.6±0.2°, 18.1±0.2°, 25.3±0.2°, 8.8±0.2°, 21.9±0.2°, 11.5±0.2°;

[0198] The X-ray powder diffraction pattern of the sulfate salt form II comprises one or more diffraction peaks located at 2θ of 15.5±0.2°, 11.1±0.2°, 8.9±0.2°, 19.3±0.2°, 22.3±0.2°, 23.6±0.2°, 17.4±0.2°, 27.3±0.2°, 17.0±0.2°, 27.9±0.2°, 15.8±0.2°, 24.2±0.2°, 21.8±0.2°, 10.3±0.2°, and 20.6±0.2°, preferably, comprising 4, 5, 6, 8 or 10 diffraction peaks selected therefrom; for example,

[0199] 11.1±0.2°, 8.9±0.2°, 19.3±0.2°, 21.8±0.2°;

[0200] 15.5±0.2°, 8.9±0.2°, 22.3±0.2°, 10.3±0.2°;

[0201] 11.1±0.2°, 8.9±0.2°, 19.3±0.2°, 22.3±0.2°, 20.6±0.2°, 27.9±0.2°;

[0202] 15.5±0.2°, 11.1±0.2°, 19.3±0.2°, 22.3±0.2°, 21.8±0.2°, 10.3±0.2°;

[0203] 15.5±0.2°, 11.1±0.2°, 8.9±0.2°, 22.3±0.2°, 23.6±0.2°, 17.4±0.2°, 20.6±0.2°, 27.9±0.2°;

[0204] 15.5±0.2°, 11.1±0.2°, 8.9±0.2°, 19.3±0.2°, 23.6±0.2°, 17.4±0.2°, 10.3±0.2°, 20.6±0.2°;

[0205] 11.1±0.2°, 8.9±0.2°, 19.3±0.2°, 22.3±0.2°, 23.6±0.2°, 17.4±0.2°, 27.3±0.2°, 17.0±0.2°, 27.9±0.2°, 20.6±0.2°;

[0206] 15.5±0.2°, 8.9±0.2°, 19.3±0.2°, 22.3±0.2°, 23.6±0.2°, 17.4±0.2°, 27.3±0.2°, 17.0±0.2°, 15.8±0.2°, 10.3±0.2°;

[0207] The X-ray powder diffraction pattern of the sulfate salt form III comprises one or more diffraction peaks located at 2θ of 19.6±0.2°, 18.0±0.2°, 18.4±0.2°, 16.8±0.2°, 14.3±0.2°, 11.8±0.2°, 14.9±0.2°, 25.7±0.2°, 15.4±0.2°, 23.5±0.2°, 18.8±0.2°, 24.7±0.2°, 9.5±0.2°, 8.8±0.2°, and 11.1±0.2°, preferably, comprising 4, 5, 6, 8 or 10 diffraction peaks selected therefrom; for example,

[0208] 19.6±0.2°, 18.0±0.2°, 18.4±0.2°, 16.8±0.2°;

[0209] 19.6±0.2°, 18.0±0.2°, 18.4±0.2°, 14.3±0.2°;

[0210] 19.6±0.2°, 18.0±0.2°, 18.4±0.2°, 16.8±0.2°, 14.3±0.2°, 11.1±0.2°;

[0211] 19.6±0.2°, 18.0±0.2°, 18.4±0.2°, 16.8±0.2°, 14.3±0.2°, 8.8±0.2°;

[0212] 19.6±0.2°, 18.0±0.2°, 18.4±0.2°, 16.8±0.2°, 14.3±0.2°, 11.8±0.2°, 14.9±0.2°, 11.1±0.2°;

[0213] 19.6±0.2°, 18.0±0.2°, 18.4±0.2°, 16.8±0.2°, 14.3±0.2°, 11.8±0.2°, 14.9±0.2°, 8.8±0.2°;

[0214] 19.6±0.2°, 18.0±0.2°, 18.4±0.2°, 16.8±0.2°, 14.3±0.2°, 11.8±0.2°, 14.9±0.2°, 25.7±0.2°, 15.4±0.2°, 23.5±0.2°;

[0215] 19.6±0.2°, 18.0±0.2°, 18.4±0.2°, 16.8±0.2°, 14.3±0.2°, 11.8±0.2°, 14.9±0.2°, 25.7±0.2°, 15.4±0.2°, 18.8±0.2°;

[0216] The X-ray powder diffraction pattern of the sulfate salt form IV comprises one or more diffraction peaks located at 2θ of 19.4±0.2°, 18.9±0.2°, 15.5±0.2°, 8.8±0.2°, 18.1±0.2°, 24.9±0.2°, 17.4±0.2°, 12.3±0.2°, 26.1±0.2°, 14.5±0.2°, 22.2±0.2°, 24.3±0.2°, 21.7±0.2°, and 23.6±0.2°, preferably, comprising 4, 5, 6, 8 or 10 diffraction peaks selected therefrom; for example,

[0217] 19.4±0.2°, 18.9±0.2°, 15.5±0.2°, 8.8±0.2°;

[0218] 19.4±0.2°, 18.9±0.2°, 15.5±0.2°, 18.1±0.2°;

[0219] 19.4±0.2°, 18.9±0.2°, 15.5±0.2°, 8.8±0.2°, 18.1±0.2°, 23.6±0.2°;

[0220] 19.4±0.2°, 18.9±0.2°, 15.5±0.2°, 8.8±0.2°, 18.1±0.2°, 21.7±0.2°;

[0221] 19.4±0.2°, 18.9±0.2°, 15.5±0.2°, 8.8±0.2°, 18.1±0.2°, 24.9±0.2°, 17.4±0.2°, 23.6±0.2°;

[0222] 19.4±0.2°, 18.9±0.2°, 15.5±0.2°, 8.8±0.2°, 18.1±0.2°, 24.9±0.2°, 17.4±0.2°, 12.3±0.2°, 26.1±0.2°, 14.5±0.2°;

[0223] 19.4±0.2°, 18.9±0.2°, 15.5±0.2°, 8.8±0.2°, 18.1±0.2°, 24.9±0.2°, 17.4±0.2°, 12.3±0.2°, 26.1±0.2°, 24.3±0.2°.

[0224] In a further preferred embodiment of the present invention, the isethionate salt form I of compound P-4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-7-(6-amino-3-chloro-2-fluorophenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one, using Cu-Kα radiation, the X-ray characteristic diffraction peaks expressed in 2θ angles and interplanar spacing d values ​​are shown in Table 1.

[0225] Table 1

[0226]

[0227] The isethionate salt form I of the compound P-4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-7-(6-amino-3-chloro-2-fluorophenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one shown in Example 13-1 of the present invention has an X-ray powder diffraction pattern substantially as shown in Figure 1; a DSC pattern substantially as shown in Figure 2; and a TGA pattern substantially as shown in Figure 3.

[0228] In a further preferred embodiment of the present invention, the isethionate salt form II of the compound P-4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-7-(6-amino-3-chloro-2-fluorophenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one shown in Example 13-1, using Cu-Kα radiation, the X-ray characteristic diffraction peaks expressed in 2θ angles and interplanar spacing d values ​​are shown in Table 2.

[0229] Table 2

[0230]

[0231]

[0232] The isethionate salt form II of the compound P-4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-7-(6-amino-3-chloro-2-fluorophenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one shown in Example 13-1 of the present invention has an X-ray powder diffraction pattern substantially as shown in Figure 4; a DSC pattern substantially as shown in Figure 5; and a TGA pattern substantially as shown in Figure 6.

[0233] In a further preferred embodiment of the present invention, the isethionate salt form III of the compound P-4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-7-(6-amino-3-chloro-2-fluorophenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one shown in Example 13-1, using Cu-Kα radiation, the X-ray characteristic diffraction peaks expressed in 2θ angles and interplanar spacing d values ​​are shown in Table 3.

[0234] Table 3

[0235]

[0236]

[0237] The isethionate salt form III of the compound P-4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-7-(6-amino-3-chloro-2-fluorophenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one shown in Example 13-1 of the present invention has an X-ray powder diffraction pattern substantially as shown in Figure 7; a DSC pattern substantially as shown in Figure 8; and a TGA pattern substantially as shown in Figure 9.

[0238] In a further preferred embodiment of the present invention, the sulfate salt crystalline form I of the compound P-4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-7-(6-amino-3-chloro-2-fluorophenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one shown in Example 13-1, using Cu-Kα radiation, the X-ray characteristic diffraction peaks expressed in 2θ angles and interplanar spacing d values ​​are shown in Table 4.

[0239] Table 4

[0240]

[0241] The sulfate salt crystalline form I of the compound P-4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-7-(6-amino-3-chloro-2-fluorophenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one shown in Example 13-1 of the present invention has an X-ray powder diffraction pattern substantially as shown in Figure 10.

[0242] In a further preferred embodiment of the present invention, the sulfate salt form II of the compound P-4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-7-(6-amino-3-chloro-2-fluorophenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one shown in Example 13-1, using Cu-Kα radiation, the X-ray characteristic diffraction peaks expressed in 2θ angles and interplanar spacing d values ​​are shown in Table 5.

[0243] Table 5

[0244]

[0245]

[0246] The sulfate salt crystalline form II of the compound P-4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-7-(6-amino-3-chloro-2-fluorophenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one shown in Example 13-1 of the present invention has an X-ray powder diffraction pattern substantially as shown in Figure 11.

[0247] In a further preferred embodiment of the present invention, the sulfate salt form III of the compound P-4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-7-(6-amino-3-chloro-2-fluorophenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one shown in Example 13-1 uses Cu-Kα radiation, and the X-ray characteristic diffraction peaks expressed in 2θ angles and interplanar spacing d values ​​are shown in Table 6.

[0248] Table 6

[0249]

[0250] The sulfate salt crystalline form III of the compound P-4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-7-(6-amino-3-chloro-2-fluorophenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one shown in Example 13-1 of the present invention has an X-ray powder diffraction pattern substantially as shown in Figure 12.

[0251] In a further preferred embodiment of the present invention, the sulfate salt form IV of the compound P-4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-7-(6-amino-3-chloro-2-fluorophenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one shown in Example 13-1, using Cu-Kα radiation, the X-ray characteristic diffraction peaks expressed in 2θ angles and interplanar spacing d values ​​are shown in Table 7.

[0252] Table 7

[0253]

[0254] The sulfate salt crystalline form IV of the compound P-4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-7-(6-amino-3-chloro-2-fluorophenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one shown in Example 13-1 of the present invention has an X-ray powder diffraction pattern substantially as shown in Figure 13.

[0255] In a further preferred embodiment of the present invention, the 2θ error between the top ten diffraction peak positions with the relative peak intensity in the X-ray powder diffraction pattern of the isethionate salt form I and the diffraction peak at the corresponding position in Figure 1 is ±0.2° to ±0.5°, preferably ±0.2° to ±0.3°, and most preferably ±0.2°;

[0256] The 2θ error between the top ten diffraction peak positions with the highest relative peak intensity in the X-ray powder diffraction pattern of the isethionate salt form II and the diffraction peak at the corresponding position in FIG4 is ±0.2° to ±0.5°, preferably ±0.2° to ±0.3°, and most preferably ±0.2°;

[0257] The 2θ error between the top ten diffraction peak positions with the highest relative peak intensity in the X-ray powder diffraction pattern of the isethionate salt form III and the diffraction peak at the corresponding position in FIG7 is ±0.2° to ±0.5°, preferably ±0.2° to ±0.3°, and most preferably ±0.2°;

[0258] The 2θ error between the top ten diffraction peak positions with the highest relative peak intensity in the X-ray powder diffraction pattern of the sulfate salt form I and the diffraction peak at the corresponding position in FIG10 is ±0.2° to ±0.5°, preferably ±0.2° to ±0.3°, and most preferably ±0.2°;

[0259] The 2θ error between the top ten diffraction peak positions with the highest relative peak intensity in the X-ray powder diffraction pattern of the sulfate salt form II and the diffraction peak at the corresponding position in FIG11 is ±0.2° to ±0.5°, preferably ±0.2° to ±0.3°, and most preferably ±0.2°;

[0260] The 2θ error between the top ten diffraction peak positions with the highest relative peak intensity in the X-ray powder diffraction pattern of the sulfate salt form III and the diffraction peak at the corresponding position in FIG12 is ±0.2° to ±0.5°, preferably ±0.2° to ±0.3°, and most preferably ±0.2°;

[0261] The 2θ error between the top ten diffraction peak positions with the highest relative peak intensity in the X-ray powder diffraction pattern of the sulfate salt form IV and the diffraction peak at the corresponding position in FIG13 is ±0.2° to ±0.5°, preferably ±0.2° to ±0.3°, and most preferably ±0.2°;

[0262] In a further preferred embodiment of the present invention, the acid salt of the compound is characterized in that the acid salt crystal form is a hydrate or anhydrous. When the acid salt crystal form is a hydrate, the number of water is 0.2-3, preferably 0.2, 0.5, 1, 1.5, 2, 2.5 or 3, more preferably 0.5, 1, 2 or 3; further, the water in the hydrate is pipeline water or crystal water or a combination of the two.

[0263] In a further preferred embodiment of the present invention, the method for preparing the acid salt comprises the following steps:

[0264] 1) Weigh an appropriate amount of free base and dissolve it in a solvent;

[0265] 2) adding an appropriate amount of acid and stirring; the amount of acid is preferably 1.2 equivalents;

[0266] 3) Rapidly centrifuge or allow to stand to obtain a salt of the compound;

[0267] The solvent is an organic solvent, preferably at least one of ethanol, 2-methyltetrahydrofuran, toluene, isopropyl acetate, tert-butanol, n-butanol, tetrahydrofuran, acetone, 2-butanone, ethyl acetate or 1,4-dioxane;

[0268] The acid is selected from the group consisting of hydrochloric acid, sulfuric acid, nitric acid, hydrobromic acid, hydrofluoric acid, hydroiodic acid, phosphoric acid, 2,5-dihydroxybenzoic acid, 1-hydroxy-2-naphthoic acid, acetic acid, ethanesulfonic acid, dichloroacetic acid, trichloroacetic acid, acetohydroxamic acid, adipic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, benzoic acid, 4-acetamidobenzoic acid, 4-aminobenzoic acid, capric acid, hexanoic acid, caprylic acid, cinnamic acid, citric acid, cyclohexanesulfamic acid, camphorsulfonic acid, aspartic acid, camphoric acid, gluconic acid, glucuronic acid, glutamic acid, isoascorbic acid, lactic acid, malic acid, mandelic acid, pyroglutamic acid, tartaric acid, dodecylsulfuric acid, dibenzoyltartaric acid, ethane-1,2-disulfonic acid , ethanesulfonic acid, formic acid, fumaric acid, galactonic acid, gentisic acid, glutaric acid, 2-ketoglutaric acid, glycolic acid, hippuric acid, isethionic acid, lactobionic acid, ascorbic acid, aspartic acid, lauric acid, camphoric acid, maleic acid, malonic acid, methanesulfonic acid, 1,5-naphthalenedisulfonic acid, naphthalene-2-sulfonic acid, nicotinic acid, oleic acid, orotic acid, oxalic acid, palmitic acid, pamoic acid, propionic acid, salicylic acid, 4-aminosalicylic acid, sebacic acid, stearic acid, succinic acid, thiocyanic acid, undecylenic acid, trifluoroacetic acid, benzenesulfonic acid, p-toluenesulfonic acid or L-malic acid; preferably hydrochloric acid, phosphoric acid, ethanesulfonic acid, benzenesulfonic acid, methanesulfonic acid, fumaric acid, isethionic acid, oxalic acid or hydrobromic acid.

[0269] In a further preferred embodiment of the present invention, the method for preparing the acid salt of the compound and its crystal form comprises the following steps:

[0270] 1) Weigh an appropriate amount of free base and dissolve it in a reaction solvent;

[0271] 2) adding an appropriate amount of acid and stirring; the amount of acid is preferably 1.2 equivalents;

[0272] 3) After centrifugal drying, a crystalline form of the acid salt of the compound is obtained;

[0273] The reaction solvent is an organic solvent, preferably at least one of ethanol, 2-methyltetrahydrofuran, n-heptane, methyl tert-butyl ether, toluene, isopropyl acetate, tert-butanol, n-butanol, tetrahydrofuran, acetone, 2-butanone, ethyl acetate or 1,4-dioxane;

[0274] The acid is selected from the group consisting of hydrochloric acid, sulfuric acid, nitric acid, hydrobromic acid, hydrofluoric acid, hydroiodic acid, phosphoric acid, 2,5-dihydroxybenzoic acid, 1-hydroxy-2-naphthoic acid, acetic acid, ethanesulfonic acid, dichloroacetic acid, trichloroacetic acid, acetohydroxamic acid, adipic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, benzoic acid, 4-acetamidobenzoic acid, 4-aminobenzoic acid, capric acid, hexanoic acid, caprylic acid, cinnamic acid, citric acid, cyclohexanesulfamic acid, camphorsulfonic acid, aspartic acid, camphoric acid, gluconic acid, glucuronic acid, glutamic acid, isoascorbic acid, lactic acid, malic acid, mandelic acid, pyroglutamic acid, tartaric acid, dodecylsulfuric acid, dibenzoyltartaric acid, ethane-1,2-disulfonic acid, ethanesulfonic acid, formic acid, fumaric acid, galactonic acid, gentisic acid , glutaric acid, 2-ketoglutaric acid, glycolic acid, hippuric acid, isethionic acid, lactobionic acid, ascorbic acid, aspartic acid, lauric acid, camphoric acid, maleic acid, malonic acid, methanesulfonic acid, 1,5-naphthalenedisulfonic acid, naphthalene-2-sulfonic acid, nicotinic acid, oleic acid, orotic acid, oxalic acid, palmitic acid, pamoic acid, propionic acid, salicylic acid, 4-aminosalicylic acid, sebacic acid, stearic acid, succinic acid, thiocyanic acid, undecylenic acid, trifluoroacetic acid, benzenesulfonic acid, p-toluenesulfonic acid or L-malic acid; preferably hydrochloric acid, phosphoric acid, ethanesulfonic acid, benzenesulfonic acid, methanesulfonic acid, fumaric acid, isethionic acid, oxalic acid or hydrobromic acid.

[0275] In a further preferred embodiment of the present invention, the method for preparing the acid salt crystalline form of the compound comprises the following steps:

[0276] 1) Weigh an appropriate amount of the salt of the compound and add an organic solvent to suspend it;

[0277] 2) stirring, and centrifugally drying to obtain a crystalline form of the acid salt of the compound;

[0278] The organic solvent is selected from at least one of ethanol, 2-methyltetrahydrofuran, n-heptane, methyl tert-butyl ether, toluene, isopropyl acetate, tert-butanol, n-butanol, tetrahydrofuran, acetone, 2-butanone, ethyl acetate and 1,4-dioxane.

[0279] In a further preferred embodiment of the present invention, the method for preparing the acid salt of the compound or its crystalline form comprises the following steps:

[0280] 1) Weigh an appropriate amount of free base and dissolve it in a reaction solvent;

[0281] 2) Add appropriate amount of acid and organic solvent and stir to dissolve;

[0282] 3) optionally, adding seed crystals;

[0283] 4) Cool, filter and separate the precipitated solid, wash with solvent and dry.

[0284] The reaction solvent used in step 1) is an organic solvent, preferably at least one of ethanol, propanol, isopropanol, 2-methyltetrahydrofuran, n-heptane, methyl tert-butyl ether, toluene, isopropyl acetate, tert-butanol, n-butanol, tetrahydrofuran, acetone, 2-butanone, ethyl acetate or 1,4-dioxane;

[0285] The acid in step 2) is selected from hydrochloric acid, sulfuric acid, nitric acid, hydrobromic acid, hydrofluoric acid, hydroiodic acid, phosphoric acid, 2,5-dihydroxybenzoic acid, 1-hydroxy-2-naphthoic acid, acetic acid, ethanesulfonic acid, dichloroacetic acid, trichloroacetic acid, acetohydroxamic acid, adipic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, benzoic acid, 4-acetamidobenzoic acid, 4-aminobenzoic acid, decanoic acid, hexanoic acid, octanoic acid, cinnamic acid, citric acid, cyclohexanesulfamic acid, camphorsulfonic acid, aspartic acid, camphoric acid, gluconic acid, glucuronic acid, glutamic acid, isoascorbic acid, lactic acid, malic acid, mandelic acid, pyroglutamic acid, tartaric acid, dodecyl sulfuric acid, dibenzoyltartaric acid, ethane-1,2- disulfonic acid, ethanesulfonic acid, formic acid, fumaric acid, galactonic acid, gentisic acid, glutaric acid, 2-ketoglutaric acid, glycolic acid, hippuric acid, isethionic acid, lactobionic acid, ascorbic acid, aspartic acid, lauric acid, camphoric acid, maleic acid, malonic acid, methanesulfonic acid, 1,5-naphthalenedisulfonic acid, naphthalene-2-sulfonic acid, nicotinic acid, oleic acid, orotic acid, oxalic acid, palmitic acid, pamoic acid, propionic acid, salicylic acid, 4-aminosalicylic acid, sebacic acid, stearic acid, succinic acid, thiocyanic acid, undecylenic acid, trifluoroacetic acid, benzenesulfonic acid, p-toluenesulfonic acid or L-malic acid; preferably hydrochloric acid, phosphoric acid, ethanesulfonic acid, benzenesulfonic acid, methanesulfonic acid, fumaric acid, isethionic acid, oxalic acid or hydrobromic acid.

[0286] The organic solvent in step 2) is selected from one or more of alcohols, ethers, ketones or esters, preferably at least one of ethanol, propanol, isopropanol, 2-methyltetrahydrofuran, n-heptane, methyl tert-butyl ether, toluene, isopropyl acetate, tert-butanol, n-butanol, tetrahydrofuran, acetone, 2-butanone, ethyl acetate or 1,4-dioxane;

[0287] The solvent in step 3) is selected from one or more of alcohols, ethers, ketones or ester solvents, preferably at least one of ethanol, propanol, isopropanol, 2-methyltetrahydrofuran, n-heptane, methyl tert-butyl ether, toluene, isopropyl acetate, tert-butanol, n-butanol, tetrahydrofuran, acetone, 2-butanone, ethyl acetate or 1,4-dioxane.

[0288] The present invention also provides a preferred embodiment and relates to a pharmaceutical composition comprising a therapeutically effective dose of an acid salt of a compound of the general formula (I) or a crystalline form thereof, and one or more pharmaceutically acceptable carriers, diluents or excipients.

[0289] The present invention further relates to the use of any acid salt of the compound of general formula (I) or its crystal form or the pharmaceutical composition in the preparation of KRAS inhibitor drugs; preferably in the preparation of KRAS G12C mutation inhibitor drugs.

[0290] In some embodiments, the pharmaceutically acceptable salts of the compounds of the present invention and their crystalline forms or compositions are used to treat Noonan syndrome, Leopard syndrome, leukemia, neuroblastoma, melanoma, breast cancer, esophageal cancer, head and neck cancer, gastric cancer, lung cancer and colon cancer; preferably non-small cell lung cancer, colon cancer, esophageal cancer, head and neck cancer.

[0291] Detailed Description of the Invention

[0292] Unless otherwise stated, the terms used in the specification and claims have the following meanings.

[0293] The term "alkyl" refers to a saturated aliphatic hydrocarbon group, which is a straight or branched chain group containing 1 to 20 carbon atoms, preferably an alkyl group containing 1 to 8 carbon atoms, more preferably an alkyl group containing 1 to 6 carbon atoms, and most preferably an alkyl group containing 1 to 3 carbon atoms. Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, n-heptyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 2,3-dimethylpentyl, 2,4-dimethylpentyl, 2,2-dimethylpentyl, 3,3-dimethylpentyl, 2-ethylpentyl, 3-Ethylpentyl, n-octyl, 2,3-dimethylhexyl, 2,4-dimethylhexyl, 2,5-dimethylhexyl, 2,2-dimethylhexyl, 3,3-dimethylhexyl, 4,4-dimethylhexyl, 2-ethylhexyl, 3-ethylhexyl, 4-ethylhexyl, 2-methyl-2-ethylpentyl, 2-methyl-3-ethylpentyl, n-nonyl, 2-methyl-2-ethylhexyl, 2-methyl-3-ethylhexyl, 2,2-diethylpentyl, n-decyl, 3,3-diethylhexyl, 2,2-diethylhexyl, and various branched-chain isomers thereof. More preferred are lower alkyl groups containing 1 to 6 carbon atoms, non-limiting examples of which include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, and the like. The alkyl group may be substituted or unsubstituted. When substituted, the substituent may be substituted at any available point of attachment. The substituent is preferably one or more of the following groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, oxo, carboxyl or carboxylate groups. Methyl, ethyl, isopropyl, tert-butyl, haloalkyl, deuterated alkyl, alkoxy-substituted alkyl and hydroxy-substituted alkyl are preferred.

[0294] The term "alkylene" refers to an alkyl group in which one hydrogen atom is further substituted, for example: "methylene" refers to -CH2-, "ethylene" refers to -(CH2)2-, "propylene" refers to -(CH2)3-, "butylene" refers to -(CH2)4-, etc. The term "alkenyl" refers to an alkyl group as defined above consisting of at least two carbon atoms and at least one carbon-carbon double bond, for example, ethenyl, 1-propenyl, 2-propenyl, 1-, 2- or 3-butenyl, etc. The alkenyl group may be substituted or unsubstituted. When substituted, the substituent is preferably one or more of the following groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, and heterocycloalkylthio.

[0295] The term "cycloalkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic hydrocarbon substituent, wherein the cycloalkyl ring contains 3 to 20 carbon atoms, preferably 3 to 12 carbon atoms, and more preferably 3 to 6 carbon atoms. Non-limiting examples of monocyclic cycloalkyls include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatrienyl, cyclooctyl, and the like; polycyclic cycloalkyls include spirocyclic, fused, and bridged cycloalkyls, preferably cyclopropyl, cyclobutyl, cyclohexyl, cyclopentyl, and cycloheptyl.

[0296] The term "spiroalkyl" refers to a polycyclic group having a carbon atom (called a spiral atom) shared between 5 to 20 monocyclic rings, which may contain one or more double bonds, but no ring has a completely conjugated π electron system. Preferably, it is 6 to 14 yuan, more preferably 7 to 10 yuan. According to the number of spiral atoms shared between the rings, the spiroalkyl is divided into a single spiroalkyl, a double spiroalkyl or a multi-spiroalkyl, preferably a single spiroalkyl and a double spiroalkyl. More preferably, it is 3 yuan / 6 yuan, 3 yuan / 5 yuan, 4 yuan / 4 yuan, 4 yuan / 5 yuan, 4 yuan / 6 yuan, 5 yuan / 5 yuan or 5 yuan / 6 yuan of single spiroalkyl. Non-limiting examples of spiroalkyl include:

[0297] wait;

[0298] It also includes spirocycloalkyl groups that share a spiro atom with a heterocycloalkyl group. Non-limiting examples include:

[0299] wait.

[0300] The term "fused cycloalkyl" refers to a 5 to 20-membered, all-carbon polycyclic group in which each ring in the system shares a pair of adjacent carbon atoms with the other rings in the system, wherein one or more rings may contain one or more double bonds, but no ring has a completely conjugated π electron system. Preferably, it is 6 to 14 members, more preferably 7 to 10 members. Depending on the number of constituent rings, it can be divided into bicyclic, tricyclic, tetracyclic or polycyclic fused cycloalkyl groups, preferably bicyclic or tricyclic, more preferably 5-membered / 5-membered or 5-membered / 6-membered bicyclic alkyl groups. Non-limiting examples of fused cycloalkyl groups include:

[0301] wait.

[0302] The term "bridged cycloalkyl" refers to a 5-20 membered, all-carbon polycyclic group in which any two rings share two carbon atoms that are not directly connected, which may contain one or more double bonds, but no ring has a completely conjugated π electron system. Preferably, it is 6-14 members, more preferably 7-10 members. Depending on the number of constituent rings, it can be classified as a bicyclic, tricyclic, tetracyclic or polycyclic bridged cycloalkyl group, preferably a bicyclic, tricyclic or tetracyclic group, more preferably a bicyclic or tricyclic group. Non-limiting examples of bridged cycloalkyl groups include:

[0303]

[0304] The cycloalkyl ring may be fused to an aryl, heteroaryl or heterocycloalkyl ring, wherein the ring attached to the parent structure is a cycloalkyl, non-limiting examples of which include indanyl, tetrahydronaphthyl, benzocycloheptanyl, etc. The cycloalkyl group may be optionally substituted or unsubstituted, and when substituted, the substituents are preferably one or more of the following groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, oxo, carboxyl or carboxylate.

[0305] The term "heterocyclyl" refers to a saturated or partially unsaturated monocyclic or polycyclic hydrocarbon substituent containing 3 to 20 ring atoms, one or more of which is selected from nitrogen, oxygen or S(O) m (wherein m is an integer from 0 to 2) heteroatoms, excluding the ring portion of -OO-, -OS- or -SS-, and the remaining ring atoms are carbon. Preferably, it contains 3 to 12 ring atoms, of which 1 to 4 are heteroatoms; more preferably, it contains 3 to 8 ring atoms; most preferably, it contains 3 to 8 ring atoms; further preferably, it is a 3-8 membered heterocyclic group containing 1-3 nitrogen atoms, optionally substituted with 1-2 oxygen atoms, sulfur atoms, or oxo groups, including nitrogen-containing monocyclic heterocyclic groups, nitrogen-containing spiro heterocyclic groups, or nitrogen-containing fused heterocyclic groups.

[0306] Non-limiting examples of monocyclic heterocyclic groups include pyrrolidinyl, imidazolidinyl, tetrahydrofuranyl, tetrahydrothiophenyl, dihydroimidazolyl, dihydrofuranyl, dihydropyrazolyl, dihydropyrrolyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl, azepanyl, 1,4-diazepanyl, pyranyl, etc., preferably pyrrolidinyl, morpholinyl, piperidinyl, azepanyl, 1,4-diazepanyl and piperazinyl. Polycyclic heterocyclic groups include spirocyclic, fused and bridged heterocyclic groups; wherein the spirocyclic, fused and bridged heterocyclic groups are optionally connected to other groups by single bonds, or further connected to other cycloalkyl, heterocyclic, aryl and heteroaryl groups through any two or more atoms on the ring.

[0307] The term "spiroheterocyclyl" refers to a polycyclic heterocyclic group in which the monocyclic rings of 5 to 20 members share one atom (called a spiro atom), wherein one or more ring atoms are selected from nitrogen, oxygen or S(O) m (wherein m is an integer 0 to 2) heteroatom, and the remaining ring atoms are carbon. It may contain one or more double bonds, but no ring has a completely conjugated π electron system. It is preferably 6 to 14 yuan, more preferably 7 to 10 yuan. According to the number of shared spiral atoms between the rings, the spiral heterocyclic group is divided into a single spiral heterocyclic group, a double spiral heterocyclic group or a multi-spiral heterocyclic group, preferably a single spiral heterocyclic group and a double spiral heterocyclic group. More preferably 3 yuan / 5 yuan, 3 yuan / 6 yuan, 4 yuan / 4 yuan, 4 yuan / 5 yuan, 4 yuan / 6 yuan, 5 yuan / 5 yuan or 5 yuan / 6 yuan single spiral heterocyclic group. Non-limiting examples of spiral heterocyclic groups include:

[0308]

[0309] wait.

[0310] The term "fused heterocyclyl" refers to a polycyclic heterocyclic group of 5 to 20 members, wherein each ring in the system shares a pair of adjacent atoms with other rings in the system, one or more rings may contain one or more double bonds, but no ring has a completely conjugated π electron system, wherein one or more ring atoms are selected from nitrogen, oxygen or S(O) m (wherein m is an integer from 0 to 2) heteroatoms, the remaining ring atoms being carbon. Preferably, it is 6 to 14 members, more preferably 7 to 10 members. According to the number of constituent rings, it can be divided into bicyclic, tricyclic, tetracyclic or polycyclic fused heterocyclic groups, preferably bicyclic or tricyclic, more preferably 5-membered / 5-membered or 5-membered / 6-membered bicyclic fused heterocyclic groups. Non-limiting examples of fused heterocyclic groups include:

[0311]

[0312]

[0313] wait.

[0314] The term "bridged heterocyclyl" refers to a 5- to 14-membered polycyclic heterocyclic group in which any two rings share two atoms that are not directly connected, which may contain one or more double bonds but no ring has a completely conjugated π electron system, wherein one or more ring atoms are selected from nitrogen, oxygen or S(O) m (wherein m is an integer from 0 to 2) heteroatoms, the remaining ring atoms being carbon. Preferably, it is 6 to 14 members, more preferably 7 to 10 members. Depending on the number of constituent rings, it can be classified as a bicyclic, tricyclic, tetracyclic or polycyclic bridged heterocyclic group, preferably a bicyclic, tricyclic or tetracyclic group, more preferably a bicyclic or tricyclic group. Non-limiting examples of bridged heterocyclic groups include:

[0315]

[0316] wait.

[0317] The heterocyclyl ring may be fused to an aryl, heteroaryl or cycloalkyl ring, wherein the ring attached to the parent structure is a heterocyclyl, non-limiting examples of which include:

[0318] wait.

[0319] The heterocyclyl group may be optionally substituted or unsubstituted. When substituted, the substituents are preferably one or more groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, oxo, carboxyl or carboxylate.

[0320] The term "aryl" refers to a 6- to 14-membered all-carbon monocyclic or fused polycyclic (i.e., rings sharing adjacent pairs of carbon atoms) group having a conjugated π electron system, preferably 6- to 12-membered, such as phenyl and naphthyl. More preferably, phenyl. The aryl ring may be fused to a heteroaryl, heterocyclic, or cycloalkyl ring, including benzo 5- to 10-membered heteroaryl, benzo 3- to 8-membered cycloalkyl, and benzo 3- to 8-membered heteroalkyl, preferably benzo 5- to 6-membered heteroaryl, benzo 3- to 6-membered cycloalkyl, and benzo 3- to 6-membered heteroalkyl, wherein the heterocyclic group is a heterocyclic group containing 1-3 nitrogen atoms, oxygen atoms, or sulfur atoms; or further comprises a three-membered nitrogen-containing fused ring containing a benzene ring.

[0321] Wherein the ring connecting to the parent structure is an aryl ring, non-limiting examples of which include:

[0322]

[0323] wait.

[0324] The aryl group may be substituted or unsubstituted. When substituted, the substituents are preferably one or more groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, carboxyl or carboxylate.

[0325] The term "heteroaryl" refers to a heteroaromatic system containing 1 to 4 heteroatoms and 5 to 14 ring atoms, wherein the heteroatoms are selected from oxygen, sulfur and nitrogen. The heteroaryl group is preferably 5 to 12-membered, more preferably 5-membered or 6-membered, such as imidazolyl, furyl, thienyl, thiazolyl, pyrazolyl, oxazolyl, pyrrolyl, triazolyl, tetrazolyl, pyridyl, pyrimidinyl, thiadiazole, pyrazinyl, etc., preferably triazolyl, thienyl, imidazolyl, pyrazolyl, oxazolyl, pyrimidinyl or thiazolyl; more preferably pyrazolyl, pyrrolyl and oxazolyl. The heteroaryl ring can be fused to an aryl, heterocyclyl or cycloalkyl ring, wherein the ring connected to the parent structure is a heteroaryl ring, non-limiting examples of which include:

[0326]

[0327] wait.

[0328] The heteroaryl group may be optionally substituted or unsubstituted, and when substituted, the substituents are preferably one or more groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, carboxyl or carboxylate.

[0329] The term "alkoxy" refers to -O-(alkyl) and -O-(unsubstituted cycloalkyl), wherein the definition of alkyl is as described above, preferably an alkyl containing 1 to 8 carbon atoms, more preferably an alkyl containing 1 to 6 carbon atoms, and most preferably an alkyl containing 1 to 3 carbon atoms. Non-limiting examples of alkoxy include: methoxy, ethoxy, propoxy, butoxy, cyclopropyloxy, cyclobutyloxy, cyclopentyloxy, cyclohexyloxy. Alkoxy can be optionally substituted or unsubstituted, and when substituted, the substituent is preferably one or more of the following groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, sulfhydryl, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio, carboxyl or carboxylate.

[0330] The term "alkylthio" refers to-S-(alkyl) and-S-(unsubstituted cycloalkyl), wherein the definition of alkyl is as described above. Preferably, the alkyl group containing 1 to 8 carbon atoms is preferably, more preferably, the alkyl group containing 1 to 6 carbon atoms is most preferably, the alkyl group containing 1 to 3 carbon atoms. The limiting examples of alkylthio include: methylthio, ethylthio, propylthio, butylthio, cyclopropylthio, cyclobutylthio, cyclopentylthio, cyclohexylthio. Alkylthio can be optionally substituted or unsubstituted, and when substituted, substituent group is preferably one or more following groups, which are independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, sulfydryl, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio, carboxyl or carboxylate.

[0331] "Alkylthio-alkyl" refers to an alkylthio group attached to an alkyl group, wherein alkyl and alkylthio are as defined above.

[0332] "Alkylaminocarbonyl" refers to an (alkyl)-NC(O)- group in which alkyl is as previously described.

[0333] "Haloalkyl" refers to an alkyl group substituted with one or more halogens, wherein alkyl is as defined above.

[0334] "Haloalkoxy" refers to an alkoxy group substituted with one or more halogens, wherein alkoxy is as defined above.

[0335] "Haloalkoxy" refers to an alkylthio group substituted with one or more halogens, wherein alkylthio is as defined above.

[0336] "Hydroxyalkyl" refers to an alkyl group substituted with a hydroxy group, wherein alkyl is as defined above.

[0337] "Alkenyl" refers to a chain alkenyl group, also known as an alkene group, preferably an alkyl group containing 2 to 8 carbon atoms, more preferably an alkyl group containing 2 to 6 carbon atoms, and most preferably an alkyl group containing 2 to 3 carbon atoms. The alkenyl group may be further substituted with other related groups, such as alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio, carboxyl or carboxylate groups.

[0338] "Alkynyl" refers to (CH≡C-), preferably an alkyl group containing 2 to 8 carbon atoms, more preferably an alkyl group containing 2 to 6 carbon atoms, and most preferably an alkyl group containing 2 to 3 carbon atoms. The alkynyl group may be further substituted with other related groups, such as alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio, carboxyl or carboxylate.

[0339] The term "alkenylcarbonyl" refers to -C(O)-(alkenyl), wherein the definition of alkenyl is as described above. Non-limiting examples of alkenylcarbonyl include: vinylcarbonyl, propenylcarbonyl, butenylcarbonyl. Alkenylcarbonyl can be optionally substituted or unsubstituted, and when substituted, the substituent is preferably one or more of the following groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, sulfhydryl, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio, carboxyl or carboxylate.

[0340] "Hydroxy" refers to an -OH group.

[0341] "Halogen" refers to fluorine, chlorine, bromine or iodine.

[0342] "Amino" refers to -NH2.

[0343] "Cyano" refers to -CN.

[0344] "Nitro" refers to -NO2.

[0345] "Carbonyl" refers to -C(O)-.

[0346] "Carboxyl" refers to -C(O)OH.

[0347] "THF" refers to tetrahydrofuran.

[0348] "EtOAc" refers to ethyl acetate.

[0349] "MeOH" refers to methanol.

[0350] "DMF" refers to N,N-dimethylformamide.

[0351] "DIPEA" refers to diisopropylethylamine.

[0352] "TFA" refers to trifluoroacetic acid.

[0353] "MeCN" refers to acetylene glycol.

[0354] "DMA" refers to N,N-dimethylacetamide.

[0355] "Et2O" refers to diethyl ether.

[0356] "DCE" refers to 1,2-dichloroethane.

[0357] "DIPEA" refers to N,N-diisopropylethylamine.

[0358] "NBS" refers to N-bromosuccinimide.

[0359] "NIS" refers to N-iodosuccinimide.

[0360] "Cbz-Cl" refers to benzyl chloroformate.

[0361] "Pd2(dba)3" refers to tris(dibenzylideneacetone)dipalladium.

[0362] "Dppf" refers to 1,1'-bis(diphenylphosphino)ferrocene.

[0363] "HATU" refers to 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethyluronium hexafluorophosphate.

[0364] "KHMDS" refers to potassium hexamethyldisilazide.

[0365] "LiHMDS" refers to lithium bis(trimethylsilylamide).

[0366] "MeLi" refers to methyllithium.

[0367] "n-BuLi" refers to n-butyllithium.

[0368] "NaBH(OAc)3" refers to sodium triacetoxyborohydride.

[0369] Different expressions such as “X is selected from A, B, or C”, “X is selected from A, B and C”, “X is A, B or C”, and “X is A, B and C” all express the same meaning, that is, X can be any one or more of A, B, and C.

[0370] The hydrogen atoms described in the present invention can all be replaced by their isotope deuterium, and any hydrogen atom in the example compounds of the present invention can also be replaced by a deuterium atom.

[0371] "Optional" or "optionally" means that the subsequently described event or circumstance may but need not occur, and that the description includes instances where the event or circumstance occurs and instances where it does not. For example, "a heterocyclic group optionally substituted with an alkyl group" means that the alkyl group may but need not be present, and that the description includes instances where the heterocyclic group is substituted with an alkyl group and instances where the heterocyclic group is not substituted with an alkyl group.

[0372] "Substituted" means that one or more hydrogen atoms, preferably up to 5, more preferably 1 to 3 hydrogen atoms, in a group are replaced independently of one another by a corresponding number of substituents. It goes without saying that the substituents are only in their possible chemical positions, and a person skilled in the art can determine (by experiment or theory) which substitutions are possible or impossible without undue effort. For example, an amino or hydroxyl group with free hydrogen may be unstable when combined with a carbon atom with an unsaturated (e.g., olefinic) bond.

[0373] A "pharmaceutical composition" refers to a mixture containing one or more compounds described herein, or their physiologically / pharmaceutically acceptable salts or prodrugs, together with other chemical components, as well as other components such as physiologically / pharmaceutically acceptable carriers and excipients. The purpose of a pharmaceutical composition is to facilitate administration to an organism, facilitating absorption of the active ingredient and thereby exerting its biological activity.

[0374] "Pharmaceutically acceptable salts" refer to salts of the compounds of the present invention that are safe and effective when used in mammals and have the desired biological activity. BRIEF DESCRIPTION OF THE DRAWINGS

[0375] Figure 1 is an XRPD diagram of P-4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-7-(6-amino-3-chloro-2-fluorophenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one isethionate salt Form I.

[0376] Figure 2 is a DSC diagram of P-4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-7-(6-amino-3-chloro-2-fluorophenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one isethionate salt Form I.

[0377] Figure 3 is a TGA chart of P-4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-7-(6-amino-3-chloro-2-fluorophenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one isethionate salt Form I.

[0378] Figure 4 is an XRPD diagram of P-4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-7-(6-amino-3-chloro-2-fluorophenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one isethionate salt Form II.

[0379] Figure 5 is a DSC chart of P-4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-7-(6-amino-3-chloro-2-fluorophenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one isethionate salt Form II.

[0380] Figure 6 is a TGA chart of P-4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-7-(6-amino-3-chloro-2-fluorophenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one isethionate salt Form II.

[0381] Figure 7 is an XRPD diagram of P-4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-7-(6-amino-3-chloro-2-fluorophenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one isethionate salt Form III.

[0382] Figure 8 is a DSC chart of P-4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-7-(6-amino-3-chloro-2-fluorophenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one isethionate salt Form III.

[0383] Figure 9 is a TGA chart of P-4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-7-(6-amino-3-chloro-2-fluorophenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one isethionate salt Form III.

[0384] Figure 10 is an XRPD diagram of P-4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-7-(6-amino-3-chloro-2-fluorophenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one sulfate salt Form I.

[0385] Figure 11 is an XRPD diagram of P-4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-7-(6-amino-3-chloro-2-fluorophenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one sulfate salt Form II.

[0386] Figure 12 is an XRPD diagram of P-4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-7-(6-amino-3-chloro-2-fluorophenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one sulfate salt Form III.

[0387] Figure 13 is an XRPD diagram of P-4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-7-(6-amino-3-chloro-2-fluorophenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one sulfate salt Form IV. DETAILED DESCRIPTION

[0388] The present invention is further described below with reference to the following examples, but these examples are not intended to limit the scope of the present invention.

[0389] 1. Preparation of Compounds

[0390] Example

[0391] The structures of the compounds of the present invention are determined by nuclear magnetic resonance (NMR) and / or liquid chromatography-mass spectrometry (LC-MS). NMR chemical shifts (δ) are given in parts per million (ppm). NMR measurements were performed using a Bruker AVANCE-400 NMR spectrometer, using deuterated dimethyl sulfoxide (DMSO-d6), deuterated methanol (CD3OD), and deuterated chloroform (CDCl3) as the solvents, with tetramethylsilane (TMS) as the internal standard.

[0392] Liquid chromatography-mass spectrometry (LC-MS) was performed on an Agilent 1200 Infinity Series mass spectrometer. HPLC was performed on an Agilent 1200DAD high-pressure liquid chromatograph (Sunfire C18 150 × 4.6 mm column) and a Waters 2695-2996 high-pressure liquid chromatograph (Gimini C 18 150×4.6mm chromatographic column).

[0393] Thin layer chromatography silica gel plates use Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plates. The specifications used for TLC are 0.15mm-0.20mm, and the specifications used for thin layer chromatography separation and purification products are 0.4mm-0.5mm. Column chromatography generally uses Yantai Huanghai silica gel 200-300 mesh silica gel as the carrier.

[0394] The starting materials in the examples of the present invention are known and can be purchased commercially, or can be synthesized using or according to methods known in the art.

[0395] Unless otherwise specified, all reactions of the present invention are carried out under continuous magnetic stirring in a dry nitrogen or argon atmosphere, with dry solvents and reaction temperatures in degrees Celsius.

[0396] Example 1

[0397] 4-((S)-4-Acryloyl-2-methylpiperazin-1-yl)-6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one

[0398]

[0399] Step 1: Preparation of 4-chloro-2-(prop-1-en-2-yl)pyridin-3-amine

[0400]

[0401] 2,4-Dichloropyridin-3-amine (4.5 g, 27.78 mmol), 4,4,5,5-tetramethyl-2-(prop-1-en-2-yl)-1,3,2-dioxaborolane (5.13 g, 30.56 mmol), potassium carbonate (11.5 g, 83.34 mmol), and Pd(PPh3)4 were added to dioxane (120 mL). The reaction mixture was mixed thoroughly and stirred overnight in an oil bath at 100°C. The mixture was concentrated under reduced pressure, and the resulting crude product was purified by flash silica gel column chromatography to obtain the target compound, 4-chloro-2-(prop-1-en-2-yl)pyridin-3-amine, as a colorless oil (4.5 g, 96% yield).

[0402] MS m / z(ESI):169.1[M+H] + .

[0403] Step 2: Preparation of 4-(methylthio)-2-(prop-1-en-2-yl)pyridin-3-amine

[0404]

[0405] 4-Chloro-2-(prop-1-en-2-yl)pyridin-3-amine (2 g, 11.9 mmol) and sodium methyl mercaptan (10 mL, 20% aqueous solution) were added to dioxane (3 mL). The reaction solution was evenly mixed and reacted at 100° C. for 2 days. The mixture was cooled to room temperature and concentrated under reduced pressure. The crude product was separated and purified by flash silica gel column chromatography to give compound 4-(methylthio)-2-(prop-1-en-2-yl)pyridin-3-amine as a light yellow liquid (1.7 g, yield 79%).

[0406] MS m / z(ESI):181.2[M+H] + .

[0407] Step 3: Preparation of 2-isopropyl-4-(methylthio)pyridin-3-amine

[0408]

[0409] Methanol (50 mL) was added to 4-(methylthio)-2-(prop-1-en-2-yl)pyridin-3-amine (2 g, 11.11 mmol) and Pd / C (4 g). The reaction mixture was mixed thoroughly, allowed to react overnight at room temperature, and then concentrated under reduced pressure. The resulting crude product was added to a solution of methanol (5 mL), N,N-diisopropylethylamine (0.5 mL), and acrylonitrile (1 mL) and allowed to react at room temperature for 2 hours. The mixture was concentrated under reduced pressure and the resulting crude product was purified by flash silica gel column chromatography to afford 2-isopropyl-4-(methylthio)pyridin-3-amine as a colorless liquid (500 mg, 25% yield).

[0410] MS m / z(ESI):183.2[M+H] + .

[0411] Step 4: Preparation of 2,6-dichloro-5-fluoro-N-((2-isopropyl-4-(methylthio)pyridin-3-yl)carbamoyl)nicotinamide

[0412]

[0413] THF (10 mL) was added to 2,6-dichloro-5-fluoronicotinamide (500 mg, 2.44 mmol) and oxalyl chloride (1.32 mL, 2.54 mmol). The reaction mixture was uniformly mixed and reacted at 60°C for 3 hours. The reaction temperature was lowered to room temperature, and triethylamine (680 mg, 6.6 mmol) and 2-isopropyl-4-(methylthio)pyridin-3-amine (400 mg, 2.2 mmol) were added. The reaction was allowed to react at room temperature for 1 hour. The mixture was concentrated under reduced pressure, and the crude product was separated and purified by flash silica gel column chromatography to give 2,6-dichloro-5-fluoro-N-((2-isopropyl-4-(methylthio)pyridin-3-yl)carbamoyl)nicotinamide as a white solid (800 mg, 87% yield).

[0414] MS m / z(ESI):417.1[M+H] + .

[0415] Step 5: Preparation of 7-chloro-6-fluoro-4-hydroxy-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one

[0416]

[0417] 2,6-Dichloro-5-fluoro-N-((2-isopropyl-4-(methylthio)pyridin-3-yl)carbamoyl)nicotinamide (800 mg, 1.92 mmol) was added to THF (20 mL). After the reaction solution was evenly mixed, KHMDS (4.8 mL, 4.8 mmol) was slowly added at 0°C. The reaction was allowed to react at room temperature for 1 hour, and the mixture was concentrated under reduced pressure. The crude product was separated and purified by flash silica gel column chromatography to give compound 7-chloro-6-fluoro-4-hydroxy-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one as a white solid (600 mg, yield 82%).

[0418] MS m / z(ESI):381.1[M+H] + .

[0419] Step 6: Preparation of tert-butyl (S)-4-(7-chloro-6-fluoro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)-2-carbonyl-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate

[0420]

[0421] 7-Chloro-6-fluoro-4-hydroxy-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one (300 mg, 0.79 mmol), phosphorus oxychloride (600 mg, 3.95 mmil), and DIPEA (1 g, 7.9 mmol) were added to THF (40 mL). The reaction solution was uniformly mixed and reacted at 80° C. for 1 hour. The reaction temperature was lowered to room temperature, and tert-butyl ( S)-3-methylpiperazine-1-carboxylate (240 mg, 1.19 mmol) was reacted at room temperature for 1 hour, concentrated under reduced pressure, and the resulting crude product was separated and purified by flash silica gel column chromatography to give the compound tert-butyl (S)-4-(7-chloro-6-fluoro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)-2-carbonyl-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate as a white solid (400 mg, yield 90%).

[0422] MS m / z(ESI):563.1[M+H] + .

[0423] Step 7: Preparation of (S)-4-(4-acryloyl-2-methylpiperazin-1-yl)-7-chloro-6-fluoro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one

[0424]

[0425] Tert-butyl (S)-4-(7-chloro-6-fluoro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)-2-carbonyl-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate (400 mg, 0.71 mmol) and TFA (2 mL) were added to CH2Cl2 (30 mL). The reaction solution was uniformly mixed and reacted at room temperature for 1 hour. The mixture was concentrated under reduced pressure. CH2Cl2 (20 mL) and DIPEA (0.3 mL) were added to the crude product. The reaction temperature was lowered to 0°C. Acryloyl chloride (0.1 mL) was slowly added to the reaction solution. The mixture was reacted at room temperature for 1 hour and concentrated under reduced pressure. The obtained crude product was separated and purified by flash silica gel column chromatography to obtain compound (S)-4-(4-acryloyl-2-methylpiperazin-1-yl)-7-chloro-6-fluoro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one as a yellow solid (200 mg, yield 55%).

[0426] MS m / z(ESI):517.1[M+H] + .

[0427] Step 8: Preparation of 4-((S)-4-acryloyl-2-methylpiperazin-1-yl)-6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one

[0428]

[0429] (S)-4-(4-acryloyl-2-methylpiperazin-1-yl)-7-chloro-6-fluoro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one (50 mg, 0.1 mmol), (2-fluoro-6-hydroxyphenyl)boronic acid (30 mg, 0.2 mmol), Pd(dppf)Cl2 (16 mg, 0.02 mmol), and cesium carbonate (100 mg, 0.3 mmol) were added to dixoane (1.5 mL). The reaction solution was uniformly mixed and reacted under microwave heating at 100°C for 1 hour. The reaction was concentrated under reduced pressure. CH2Cl2 (20 mL) and DIPEA (0.3 mL) were added to the crude product. The reaction temperature was lowered to 0°C. Acryloyl chloride (0.1 mL) was slowly added to the reaction solution, reacted at room temperature for 1 hour, and concentrated under reduced pressure. The crude product was separated and purified by Pre-HPLC to give the compound 4-((S)-4-acryloyl-2-methylpiperazin-1-yl)-6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one as a white solid (14 mg, yield: 24%).

[0430] MS m / z(ESI):593.1[M+H] + .

[0431] 1 H NMR(400MHz,MeOD-d4)δ8.40(d,J=5.6Hz,1H),8.22-8.27(m,1H),7.21-7.27(m,2H),6.7 9-6.88(m,1H),6.58-6.66(m,2H),6.28-6.34(m,1H),5.84(d,J=12.0Hz,1H),5.06(s,1H) ,4.43-4.59(m,2H),4.07-4.23(s,1H),3.57-3.85(m,2H),3.20-3.48(m,1H),2.79-2.85 (m,1H),2.41(s,3H),1.47(d,J=4.8Hz,3H),1.20(d,J=6.4Hz,3H),1.06(d,J=6.8Hz,3H).

[0432] Example 1-1 and Example 1-2

[0433] (P-4-((S)-4-acryloyl-2-methylpiperazin-1-yl)-6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one) and (M-4-((S)-4-acryloyl-2-methylpiperazin-1-yl)-6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one)

[0434]

[0435]

[0436] Example 1 Two axial chiral isomers, Example 1-1 and Example 1-2, were obtained by SFC separation. SFC: Chiral preparation conditions:

[0437] Instrument: SFC-150 (Thar, Waters); column type: IC 20*250 mm, 10 μm (Daicel); column pressure: 100 bar; mobile phase: CO2 / Methanol (0.2% Methanol Ammonia) = 50 / 50; flow rate: 120 g / min; detection wavelength: UV 214 nm; column temperature: 35°C

[0438] Example 1-1:

[0439] t R =1.92min

[0440] MS m / z(ESI):593.1[M+H] + .

[0441] 1 H NMR(400MHz,MeOD-d4)δ8.40(d,J=5.6Hz,1H),8.22-8.27(m,1H),7.21-7.27(m,2H),6.7 9-6.88(m,1H),6.58-6.66(m,2H),6.28-6.34(m,1H),5.84(d,J=12.0Hz,1H),5.06(s,1H) ,4.43-4.59(m,2H),4.07-4.23(s,1H),3.57-3.85(m,2H),3.20-3.48(m,1H),2.79-2.85 (m,1H),2.41(s,3H),1.47(d,J=4.8Hz,3H),1.20(d,J=6.4Hz,3H),1.06(d,J=6.8Hz,3H).

[0442] Example 1-2:

[0443] t R =2.43min

[0444] MS m / z(ESI):593.1[M+H] + .

[0445] 1 HNMR(400MHz,MeOD-d4)δ8.40(d,J=5.6Hz,1H),8.25(t,J=10.8Hz,1H),7.21-7.27(m,2H),6. 79-6.90(m,1H),6.58-6.66(m,2H),6.28-6.34(m,1H),5.83(dd,J=10.8Hz,2.0Hz,1H),5.05- 5.10(m,1H),4.41-4.57(m,2H),4.07-4.21(m,1H),3.61-3.87(m,2H),3.24-3.36(m,1H),2.7 7-2.83(m,1H),2.41(s,3H),1.46-1.49(m,3H),1.19(d,J=6.8Hz,3H),1.06(d,J=6.8Hz,3H).

[0446] Example 2

[0447] Preparation of 4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one

[0448]

[0449] Step 1: Preparation of 4,7-dichloro-6-fluoro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one

[0450]

[0451] To a solution of 7-chloro-6-fluoro-4-hydroxy-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one (200 mg, 0.526 mmol) in acetonitrile (10 mL) was added N,N-diisopropylethylamine (407 mg, 3.16 mmol) and phosphorus oxychloride (242 mg, 1.58 mmol) at room temperature. The mixture was stirred at 80°C for 1 hour. The mixture was cooled to room temperature and used directly in the next reaction.

[0452] Step 2: Preparation of tert-butyl (2R,5S)-4-(7-chloro-6-fluoro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)-2-carbonyl-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-2,5-dimethylpiperazine-1-carboxylate

[0453]

[0454] Add N,N-diisopropylethylamine (678 mg, 5.26 mmol) and tert-butyl (2R,5S)-2,5-dimethylpiperazine-1-carboxylate (224 mg, 1.005 mmol) to the reaction mixture of the previous step, and stir at room temperature for 1 hour. Water (60 mL) was added, and the mixture was extracted with ethyl acetate (40 mL × 3). The organic phase was washed with aqueous ammonium chloride solution (40 mL) and then with aqueous sodium chloride solution (30 mL). After concentration, the residue was purified by column chromatography [eluent: dichloromethane to methanol / dichloromethane from 0% to 2.2%] to give tert-butyl (2R,5S)-4-(7-chloro-6-fluoro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)-2-carbonyl-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-2,5-dimethylpiperazine-1-carboxylate (200 mg, 66% yield in 2 steps) as a yellow solid.

[0455] MS m / z(ESI):577.2[M+H] + ,579.2[M+H+2] +

[0456] Step 3: Preparation of 7-chloro-4-((2S,5R)-2,5-dimethylpiperazin-1-yl)-6-fluoro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one trifluoroacetate

[0457]

[0458] To a solution of tert-butyl (2R,5S)-4-(7-chloro-6-fluoro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)-2-carbonyl-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-2,5-dimethylpiperazine-1-carboxylate (200 mg, 0.347 mmol) in dichloromethane (6 mL) was added trifluoroacetic acid (1.2 mL). The mixture was stirred at room temperature for 1.5 hours. The reaction mixture was concentrated at low temperature to afford 7-chloro-4-((2S,5R)-2,5-dimethylpiperazin-1-yl)-6-fluoro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one trifluoroacetate (200 mg) as a red oil, which was quickly used in the next reaction.

[0459] MS m / z(ESI):477.2[M+H] + ,479.2[M+H+2] + .

[0460] Step 4: Preparation of 4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-7-chloro-6-fluoro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one

[0461]

[0462] To a solution of 7-chloro-4-((2S,5R)-2,5-dimethylpiperazin-1-yl)-6-fluoro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one trifluoroacetate (200 mg, 0.347 mmol) in dichloromethane (15 mL) was added N,N-diisopropylethylamine (447 mg, 3.47 mmol), and acryloyl chloride (63 mg, 0.694 mmol) was added dropwise at 0°C. The mixture was stirred for 1 hour after the addition was completed. The reaction was quenched with aqueous ammonium chloride (30 mL), extracted with dichloromethane (30 mL × 3), and the dichloromethane layer was washed with saturated aqueous NaCl solution (20 mL), dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography [eluent: dichloromethane to methanol / dichloromethane from 0% to 2.5%] to give 4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-7-chloro-6-fluoro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one (130 mg, 71% yield over 2 steps) as a yellow solid.

[0463] MS m / z(ESI):530.2[M+H] + ,532.2[M+H+2] + .

[0464] Step 5: Preparation of 4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one

[0465]

[0466] 4-((2S,5R)-4-Acryloyl-2,5-dimethylpiperazin-1-yl)-7-chloro-6-fluoro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one (130 mg, 0.246 mmol), (2-fluoro-6-hydroxyphenyl)boronic acid (77 mg, 0.491 mmol), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium dichloromethane complex (40 mg, 0.0491 mmol) and cesium carbonate (240 mg, 0.738 mmol) were added to dioxane (8 mL) and water (1 mL), the atmosphere was replaced with nitrogen, and the mixture was stirred in a microwave at 100°C for 1 hour. The reaction solution was concentrated and purified by column chromatography [eluent: dichloromethane to methanol / dichloromethane from 0% to 2.5%] to give 4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one (90 mg, yield 60%) as a yellow solid.

[0467] MS m / z(ESI):606.2[M+H] + .

[0468] 1 H NMR(400MHz,MeOD-d4)δ8.40(d,J=8Hz,1H),8.29-8.18(m,1H),7.30–7.18(m,2H),6.93–6.73 (m,1H),6.70–6.56(m,2H),6.36–6.20(m,1H),5.89–5.75(m,1H),5.15–4.98(m,1H),4.63–4. 22(m,2H),4.11–3.82(m,2H),3.68–3.40(m,1H),2.88–2.65(m,1H),2.40(d,J=4Hz,3H),1.53 –1.43(m,3H),1.36(t,J=8Hz,1H),1.28(t,J=8Hz,2H),1.23–1.16(m,3H),1.10–1.01(m,3H).

[0469] Example 2-1 and Example 2-2

[0470] (P-4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one) and (M-4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one)

[0471]

[0472]

[0473] Example 2 Two axial chiral isomers, Example 2-1 and Example 2-2, were obtained by SFC separation. SFC: Chiral preparation conditions:

[0474] Instrument: SFC-150 (Thar, Waters); column type: IC 20*250mm, 10μm (Daicel); column pressure: 100 bar; mobile phase: CO2 / Methanol (0.2% Methanol Ammonia) = 60 / 40; flow rate: 100 g / min; detection wavelength: UV 214 nm; column temperature: 35°C

[0475] Example 2-1:

[0476] t R =1.99min

[0477] MS m / z(ESI):606.2[M+H] + .

[0478] 1 H NMR(400MHz,MeOD-d4)δ8.40(d,J=8Hz,1H),8.29-8.18(m,1H),7.30–7.18(m,2H),6.93–6.73 (m,1H),6.70–6.56(m,2H),6.36–6.20(m,1H),5.89–5.75(m,1H),5.15–4.98(m,1H),4.63–4. 22(m,2H),4.11–3.82(m,2H),3.68–3.40(m,1H),2.88–2.65(m,1H),2.40(d,J=4Hz,3H),1.53 –1.43(m,3H),1.36(t,J=8Hz,1H),1.28(t,J=8Hz,2H),1.23–1.16(m,3H),1.10–1.01(m,3H).

[0479] Example 2-2:

[0480] t R =2.87min

[0481] MS m / z(ESI):606.2[M+H] + .

[0482] 1 H NMR(400MHz,MeOD-d4)δ8.40(d,J=8Hz,1H),8.27-8.18(m,1H),7.30–7.19(m,2H),6.94–6.74(m,1 H),6.70–6.56(m,2H),6.36–6.20(d,J=16Hz,1H),5.90–5.75(m,1H),5.14–4.98(m,1H),4.63–4.22 (m,2H),4.12–3.82(m,2H),3.68–3.41(m,1H),2.87–2.65(m,1H),2.40(d,J=4Hz,3H),1.53–1.42(m ,3H),1.36(t,J=8Hz,1H),1.28(t,J=8Hz,2H),1.23–1.16(d,J=4Hz,3H),1.10–1.01(d,J=4Hz,3H).

[0483] Example 3

[0484] 4-((S)-4-Acryloyl-2-methylpiperazin-1-yl)-6-chloro-7-(2-fluoro-6-hydroxyphenyl)-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one)

[0485]

[0486] Preparation of 4-((S)-4-acryloyl-2-methylpiperazin-1-yl)-6-chloro-7-(2-fluoro-6-hydroxyphenyl)-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one) Reference Example 1.

[0487] MS m / z(ESI):609.1[M+H] + .

[0488] Example 4

[0489] 4-((S)-4-Acryloyl-2-methylpiperazin-1-yl)-6-fluoro-7-(2-fluoro-6-(methylthio)phenyl)-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one)

[0490]

[0491] Preparation of 4-((S)-4-acryloyl-2-methylpiperazin-1-yl)-6-fluoro-7-(2-fluoro-6-(methylthio)phenyl)-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one) Reference Example 1.

[0492] MS m / z(ESI):622.8[M+H] + .

[0493] Example 5

[0494] Preparation of 4-((S)-4-acryloyl-2-methylpiperazin-1-yl)-7-(2-amino-6-fluorophenyl)-6-fluoro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one

[0495]

[0496] Step 1: Preparation of N-(4-chloro-3-fluorophenyl)-2,2,2-trifluoroacetamide

[0497]

[0498] Dissolve 4-chloro-3-fluoroaniline (1.45 g, 0.01 mol) in THF (150 mL), add Na₂CO₃ (3.18 g, 0.03 mol), and under nitrogen, cool to 0°C. Add trifluoroacetic anhydride (4.2 mL, 0.03 mol) dropwise. Stir at room temperature for 10 hours. Add the reaction mixture to water (150 mL). Extract three times with ethyl acetate (100 mL). Combine the organic layers, dry over anhydrous sodium sulfate, and concentrate to obtain the crude product. Purify by column chromatography (PE / EA = 5:1) to obtain the desired product, N-(4-chloro-3-fluorophenyl)-2,2,2-trifluoroacetamide (2.3 g, 95% yield), as a white solid.

[0499] 1 H NMR (400MHz, MeOD-d4) δ7.70 (dd, J=11.1, 2.0Hz, 1H), 7.49–7.40 (m, 2H);

[0500] 19F NMR(376MHz,MeOD-d4)δ-77.17(s);

[0501] MS m / z(ESI):242.1[M+H] + .

[0502] Step 2: Preparation of (6-amino-3-chloro-2-fluorophenyl)boronic acid

[0503]

[0504] Dissolve N-(4-chloro-3-fluorophenyl)-2,2,2-trifluoroacetamide (2.3 g, 9.5 mmol) in THF (40 mL). Under nitrogen, cool to -78°C and add n-BuLi (7.9 mL, 19.0 mmol, 2.4 M) dropwise. Stir at -50°C for 50 minutes. Cool the reaction mixture to -78°C and add triisopropyl borate (2.3 g, 9.5 mmol) (4.8 mL, 20.9 mmol) dropwise. Stir at the same temperature for 20 minutes. Remove the dry ice bath and stir at room temperature for 2 hours. Then, cool the reaction mixture to 0°C and add dilute hydrochloric acid (19 mL, 1 M). Warm to 40°C and stir for 1 hour. Extract three times with ethyl acetate (100 mL). The organic layers were combined, dried over anhydrous sodium sulfate, and concentrated to give a crude product, which was purified by column chromatography (PE / EA=4:1) to give the target product (6-amino-3-chloro-2-fluorophenyl)boronic acid (1.1 g, yield 56%) as a gray solid.

[0505] MS m / z(ESI):190.0[M+H] + .

[0506] Step 3: Preparation of (2-amino-6-fluorophenyl)boronic acid

[0507]

[0508] (6-Amino-3-chloro-2-fluorophenyl)boronic acid (100 mg, 0.53 mmol) was dissolved in MeOH (20 mL) and Pd / C (20 mg) was added. The atmosphere was replaced with hydrogen three times and stirred at 15 psi for 2 hours. The reaction was complete as determined by TLC (PE / EA 1:1). Filtration and concentration of the filtrate afforded the desired product, (2-amino-6-fluorophenyl)boronic acid, as a yellow solid (80 mg, 97% yield), which was used directly in the next step without purification.

[0509] MS m / z(ESI):156.0[M+H] + .

[0510] Step 4: Preparation of 4-((S)-4-acryloyl-2-methylpiperazin-1-yl)-7-(2-amino-6-fluorophenyl)-6-fluoro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one

[0511]

[0512] (S)-4-(4-acryloyl-2-methylpiperazin-1-yl)-7-chloro-6-fluoro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one (26 mg, 0.05 mmol), (6-amino-3-chloro-2-fluorophenyl)boronic acid (23.2 mg, 0.15 mmol), and cesium carbonate (48.87 mg, 0.15 mmol) were dissolved in dioxane / H2O (1.5 mL / 0.3 mL). The atmosphere was replaced with nitrogen for 1 minute and the reaction was carried out in a microwave at 100°C for 1 hour. After the reaction was completed, the reaction solution was concentrated and purified by column chromatography (CH2Cl2 / MeOH=20:1) to give a crude product, which was then purified by preparative HPLC to give the target product 4-((S)-4-acryloyl-2-methylpiperazin-1-yl)-7-(2-amino-6-fluorophenyl)-6-fluoro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one (7.0 mg, yield 24%) as a yellow solid.

[0513] 1 H NMR(400MHz,MeOD-d4)δ8.46(d,J=5.4Hz,1H),8.25(dd,J=21.2,12.0Hz,1H),7.27(d,J=5.5Hz,1H),7.1 1(dd,J=14.7,8.2Hz,1H),6.84(d,J=14.2Hz,1H),6.49(d,J=8.3Hz,1H),6.41–6.27(m,2H),5.83(dd,J=1 0.6,1.6Hz,1H),4.48(dd,J=52.4,11.6Hz,2H),4.30–3.83(m,2H),3.74(d,J=9.7Hz,2H),3.22(s,1H),2 .98–2.80(m,1H),2.43(d,J=0.7Hz,3H),1.56–1.40(m,3H),1.22(d,J=6.6Hz,3H),1.01(d,J=6.6Hz,3H).

[0514] 19F NMR(376MHz,MeOD-d4)δ-114.58–-114.95(m),-114.95–-115.34(m),-125.12–-126.48(m).

[0515] MS m / z(ESI):592.2[M+H] + .

[0516] Example 6

[0517] 2-(4-((S)-4-acryloyl-2-methylpiperazin-1-yl)-6-fluoro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)-2-oxyl-1,2-dihydropyridin[2,3-d]pyrimidin-7-yl)-3-fluorobenzamide

[0518]

[0519] The preparation of 2-(4-((S)-4-acryloyl-2-methylpiperazin-1-yl)-6-fluoro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)-2-oxyl-1,2-dihydropyrido[2,3-d]pyrimidin-7-yl)-3-fluorobenzamide was carried out according to Example 1.

[0520] MS m / z(ESI):619.7[M+H] + .

[0521] Example 7

[0522] 4-((S)-4-Acryloyl-2-methylpiperazin-1-yl)-7-(2-(dimethylamino)-6-fluorophenyl)-6-fluoro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyridinyl[2,3-d]pyrimidin-2(1H)-one

[0523]

[0524] The preparation of 4-((S)-4-acryloyl-2-methylpiperazin-1-yl)-7-(2-(dimethylamino)-6-fluorophenyl)-6-fluoro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyridinyl[2,3-d]pyrimidin-2(1H)-one was carried out according to Example 1.

[0525] MS m / z(ESI):619.7[M+H] + .

[0526] Example 8

[0527]

[0528] The preparation of 4-((S)-4-acryloyl-2-methylpiperazin-1-yl)-6-fluoro-7-(2-fluoro-6-(methylamino)phenyl)-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyridinyl[2,3-d]pyrimidin-2(1H)-one was carried out according to Example 1.

[0529] MS m / z(ESI):605.7[M+H] + .

[0530] Example 9

[0531] Preparation of 4-((S)-4-acryloyl-2-methylpiperazin-1-yl)-7-(6-amino-3-chloro-2-fluorophenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one

[0532]

[0533] (S)-4-(4-acryloyl-2-methylpiperazin-1-yl)-7-chloro-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one (26.7 mg, 0.05 mmol), (6-amino-3-chloro-2-fluorophenyl)boronic acid (28.4 mg, 0.15 mmol), and potassium acetate (15.0 mg, 0.15 mmol) were dissolved in dioxane / H2O (1.5 mL / 0.3 mL). The atmosphere was replaced with nitrogen for 1 minute and the reaction was carried out in a microwave at 100°C for 1 hour. After the reaction was completed, the reaction solution was concentrated and purified by column chromatography (CH2Cl2 / MeOH=20:1) to give a crude product, which was then purified by preparative HPLC to give the target product 4-((S)-4-acryloyl-2-methylpiperazin-1-yl)-7-(2-amino-6-fluorophenyl)-6-fluoro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one (3.7 mg, yield 14%) as a yellow solid.

[0534] MS m / z(ESI):642.1[M+H] + .

[0535] 1H NMR (400MHz, MeOD-d4) δ8.56–8.39(m,2H),7.24(t,J=5.3Hz,1H),7.15(dd,J=15.4,6.9H z,1H),6.84(d,J=9.9Hz,1H),6.53–6.46(m,1H),6.32(d,J=15.9Hz,1H),5.84(d,J=12.2H z,1H),4.68–4.36(m,3H),4.10(dd,J=45.7,31.6Hz,2H),3.76(s,1H),2.94(s,2H),2.42 (d,J=6.2Hz,3H),1.57–1.43(m,3H),1.22(d,J=6.7Hz,3H),1.06(dd,J=42.4,6.7Hz,3H). 19 F NMR(376MHz,MeOD)δ-117.04–-117.24(m),-117.24–-117.51(m).

[0536] Example 9-1 and Example 9-2

[0537] (P-4-((S)-4-Acryloyl-2-methylpiperazin-1-yl)-7-(2-amino-6-fluorophenyl)-6-fluoro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one) and (M-4-((S)-4-Acryloyl-2-methylpiperazin-1-yl)-7-(2-amino-6-fluorophenyl)-6-fluoro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one)

[0538]

[0539] Example 9 was resolved by SFC to obtain two axial chiral isomers, Example 9-1 and Example 9-2. SFC: Chiral preparation conditions:

[0540] Instrument: SFC-80 (Thar, Waters) column type IC 20*250mm, 10μm (Daicel) column pressure 100bar

[0541] Mobile phase: CO2 / Methanol (0.2% Methanol Ammonia) = 45 / 55; Flow rate: 80 g / min; Detection wavelength: UV: 214 nm; Column temperature: 35°C

[0542] Example 9-1:

[0543] t R =1.74min

[0544] MS m / z (ESI): 642.1 [M+H] + 。

[0545] 1 H NMR (400 MHz, MeOD-d4) δ 8.56–8.39 (m, 2H), 7.24 (t, J = 5.3 Hz, 1H), 7.15 (dd, J = 15.4, 6.9 Hz, 1H), 6.84 (d, J = 9.9 Hz, 1H), 6.53–6.46 (m, 1H), 6.32 (d, J = 15.9 Hz, 1H), 5.84 (d, J = 12.2 Hz, 1H), 4.68–4.36 (m, 3H), 4.10 (dd, J = 45.7, 31.6 Hz, 2H), 3.76 (s, 1H), 2.94 (s, 2H), 2.42 (d, J = 6.2 Hz, 3H), 1.57–1.43 (m, 3H), 1.22 (d, J = 6.7 Hz, 3H), 1.06 (dd, J = 42.4, 6.7 Hz, 3H).

[0546] 19 F NMR (376 MHz, MeOD-d4) δ -117.04–-117.24 (m), -117.24–-117.51 (m).

[0547] Example 9-2:

[0548] t R = 2.49 min

[0549] MS m / z (ESI): 642.1 [M+H] + 。

[0550] 1 H NMR (400 MHz, MeOD-d4) δ 8.56–8.39 (m, 2H), 7.27–7.10 (m, 2H), 6.84 (dd, J = 28.3, 17.7 Hz, 1H), 6.50 (d, J = 8.8 Hz, 1H), 6.32 (d, J = 16.9 Hz, 1H), 5.83 (d, J = 11.7 Hz, 1H), 4.63–4.41 (m, 2H), 4.23–4.02 (m, 1H), 3.79–3.57 (m, 2H), 3.36 (s, 2H), 2.99–2.86 (m, 1H), 2.41 (d, J = 7.6 Hz, 3H), 1.51 (d, J = 25.9 Hz, 3H), 1.21 (d, J = 6.6 Hz, 3H), 1.05 (dd, J = 44.8, 6.7 Hz, 3H).

[0551] Example 10

[0552] Preparation of 4-((S)-4-acryloyl-2-methylpiperazin-1-yl)-7-(6-amino-3-chloro-2-fluorophenyl)-6-fluoro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one

[0553]

[0554] (S)-4-(4-acryloyl-2-methylpiperazin-1-yl)-7-chloro-6-fluoro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one (26 mg, 0.05 mmol), (6-amino-3-chloro-2-fluorophenyl)boronic acid (28.4 mg, 0.15 mmol), and cesium carbonate (48.8 mg, 0.15 mmol) were dissolved in dioxane / H2O (1.5 mL / 0.3 mL). The atmosphere was replaced with nitrogen for 1 minute and the reaction was carried out in a microwave at 100°C for 1 hour. After the reaction, the reaction solution was dried and purified by column chromatography (CH2Cl2 / MeOH=20:1) to give a crude product, which was then purified by preparative HPLC to give the target product 4-((S)-4-acryloyl-2-methylpiperazin-1-yl)-7-(2-amino-6-fluorophenyl)-6-fluoro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one as a yellow solid (4.4 mg, yield 14%).

[0555] 1 H NMR(400MHz,MeOD-d4)δ8.47(d,J=5.4Hz,1H),8.38–8.24(m,1H),7.27(d,J=5.4Hz,1H),7.17(t ,J=8.6Hz,1H),6.85(d,J=14.9Hz,1H),6.49(d,J=8.9Hz,1H),6.32(d,J=16.3Hz,1H),5.84(d,J =10.5Hz,1H),4.57(d,J=23.5Hz,2H),4.42(s,1H),4.24–3.89(m,2H),3.73(dd,J=14.4,7.9Hz, 1H),2.92(s,1H),2.43(s,3H),1.54–1.40(m,3H),1.22(d,J=6.7Hz,3H),1.01(d,J=6.6Hz,3H).

[0556] 19F NMR(376MHz,MeOD-d4)δ-116.46–-116.73(m),-116.87(dd,J=39.0,8.4Hz),-126.18(dd,J=24.9,15.2Hz).

[0557] MS m / z(ESI):626.1[M+H] + .

[0558] Example 11

[0559] Preparation of 4-((S)-4-acryloyl-2-methylpiperazin-1-yl)-7-(2,3-difluoro-6-hydroxyphenyl)-6-fluoro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one

[0560]

[0561] Preparation of 4-((S)-4-acryloyl-2-methylpiperazin-1-yl)-7-(2,3-difluoro-6-hydroxyphenyl)-6-fluoro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one Reference Example 2.

[0562] MS m / z(ESI):611.1[M+H] + .

[0563] 1 H NMR(400MHz,MeOD-d4)δ8.41(d,J=5.6Hz,1H),8.32-8.25(m,1H),7.25(d,J=5.6Hz,1H),7 .20-7.13(m,1H),6.92-6.82(m,1H),6.62-6.58(m,1H),6.34-6.28(m,1H),5.83(d,J=10.4 Hz,1H),5.14-5.04(m,1H),4.64-4.42(m,2H),4.25-4.07(m,1H),3.89-3.61(m,3H),2.88- 2.77(m,1H),2.42(s,3H),1.52-1.46(m,3H),1.20(d,J=6.4Hz,3H),1.05(d,J=6.4Hz,3H).

[0564] Example 12

[0565] Preparation of (S)-4-(4-acryloyl-2-methylpiperazin-1-yl)-7-(2,6-difluorophenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one

[0566]

[0567] Preparation of (S)-4-(4-acryloyl-2-methylpiperazin-1-yl)-7-(2,6-difluorophenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one Reference Example 2.

[0568] MS m / z(ESI):611.1[M+H] + .

[0569] Example 13

[0570] Preparation of 4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-7-(6-amino-3-chloro-2-fluorophenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one

[0571]

[0572] Step 1: Preparation of 2,5,6-trichloro-N-((2-isopropyl-4-(methylthio)pyridin-3-yl)carbamoyl)nicotinamide

[0573]

[0574] Under N2 protection, 2,5,6-trichloronicotinamide (6.2 g, 27.7 mmol) was dissolved in THF (60 mL), and oxalyl chloride (15.2 mL, 31.5 mmol) (2M / L dichloromethane solution) was added dropwise at -78 ° C. The mixture was stirred at -78 ° C for 10 minutes and at 60 ° C for 3 hours. The reaction solution was cooled to 0 ° C. Triethylamine (18 mL, 111 mmol) was added dropwise, and a THF solution of 2-isopropyl-4-(methylthio)pyridin-3-amine (5 g, 27.7 mmol) was added dropwise, and stirred at room temperature for 2 hours. The reaction mixture was quenched with brine, extracted with water and ethyl acetate (3*100 mL), and the organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to give a crude product, which was purified by column chromatography (DCM / MeOH=100:1 to 70:1) to give the target product 2,5,6-trichloro-N-((2-isopropyl-4-(methylthio)pyridin-3-yl)carbamoyl)nicotinamide (8.6 g, yield 72%).

[0575] MS m / z(ESI):433.1[M+H] + ,435.1[M+H+2] + .

[0576] Step 2: Preparation of 6,7-dichloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidine-2,4(1H,3H)-dione

[0577]

[0578] 2,5,6-Trichloro-N-((2-isopropyl-4-(methylthio)pyridin-3-yl)carbamoyl)nicotinamide (10.4 g, 24.1 mmol) was dissolved in anhydrous THF (80 mL). Under nitrogen, the mixture was cooled to 0°C and KHMDS (48 mL, 48.2 mmol) was added dropwise. The mixture was stirred for 0.5 hours. The mixture was quenched with saturated aqueous ammonium chloride and extracted with water and ethyl acetate (3 x 100 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated to afford the crude product, which was purified by slurrying with ethyl acetate to afford the desired product, 6,7-dichloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidine-2,4(1H,3H)-dione (8 g, 84% yield).

[0579] MS m / z(ESI):397.1[M+H] + ,399.1[M+H+2] + .

[0580] Step 3: Preparation of 4,6,7-trichloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one

[0581]

[0582] Dissolve 6,7-dichloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidine-2,4(1H,3H)-dione (5.2 g, 13.1 mmol) in ACN (50 mL). Add DIEA (23 mL, 66 mmol) and POCl3 (3 mL, 19.7 mmol). Stir at 80°C for 0.5 h. Use directly in the next reaction.

[0583] MS m / z(ESI):415.1[M+H] + ,417.1[M+H+2] + .

[0584] Step 4: Preparation of tert-butyl (2R,5S)-4-(6,7-dichloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)-2-carbonyl-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-2,5-dimethylpiperazine-1-carboxylate

[0585]

[0586] To a solution of 4,6,7-trichloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one in acetonitrile (50 mL) was added DIEA (23 mL, 66 mmol) and tert-butyl (2R,5S)-2,5-dimethylpiperazine-1-carboxylate (6.2 g, 26.2 mmol). The mixture was stirred at room temperature for 1 hour. The mixture was quenched with water and extracted with water and ethyl acetate (3 x 100 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to give a crude product, which was purified by column chromatography (CH2Cl2 / MeOH=30:1) to give the target product, tert-butyl (2R,5S)-4-(6,7-dichloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)-2-carbonyl-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-2,5-dimethylpiperazine-1-carboxylate (6.1 g, yield 77%).

[0587] MS m / z(ESI):593.1[M+H] + ,595.1[M+H+2] + .

[0588] Step 5: Preparation of 6,7-dichloro-4-((2S,5R)-2,5-dimethylpiperazin-1-yl)-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one

[0589]

[0590] Tert-butyl (2R,5S)-4-(6,7-dichloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)-2-carbonyl-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-2,5-dimethylpiperazine-1-carboxylate (6.1 g, 10.3 mmol) was dissolved in dichloromethane (20 mL). TFA (20 mL) was added and the mixture was stirred at room temperature for 1 hour. Concentration was performed to obtain the crude target product, 6,7-dichloro-4-((2S,5R)-2,5-dimethylpiperazin-1-yl)-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one (6.1 g, 100% yield).

[0591] MS m / z(ESI):493.1[M+H] + ,495.1[M+H+2] + .

[0592] Step 6: Preparation of 4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-6,7-dichloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one

[0593]

[0594] Dissolve 6,7-dichloro-4-((2S,5R)-2,5-dimethylpiperazin-1-yl)-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one (6 g, 12.2 mmol) in dichloromethane (30 mL). Add DIEA (30 mL, 131 mmol) and acryloyl chloride (1.08 mL, 13.13 mmol). Stir at room temperature for 1 hour. Quench with water and extract with water and ethyl acetate (3 x 100 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to give a crude product, which was purified by column chromatography (CH2Cl2 / MeOH=20:1) to give the target product 4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-6,7-dichloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one (1.6 g, yield 22%).

[0595] MS m / z(ESI):547.1[M+H] + ,549.1[M+H+2] + .

[0596] Step 7: Preparation of 4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-7-(6-amino-3-chloro-2-fluorophenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one

[0597]

[0598] Under N2 protection, 4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-6,7-dichloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one (700 mg, 1.3 mmol) and (6-amino-3-chloro-2-fluorophenyl)boronic acid (380 mg, 2.6 mmol) were dissolved in a mixture of 1,4-dioxane and water (6 mL:0.3 mL), and the mixture was reacted with Pd(dppf)Cl2.DCM (100 mg, 0.1 mmol) and KOAc (400 mg, 4 mmol) under microwave evaporation at 100°C for 1 hour. The mixture was quenched with water and extracted with water and ethyl acetate (3*50 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to give a crude product, which was purified by column chromatography (CH2Cl2 / MeOH=200:1 to 80:1) to give the target product 4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-7-(6-amino-3-chloro-2-fluorophenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one (400 mg, 48% yield).

[0599] MS m / z(ESI):656.1[M+H] + ,658.1[M+H+2] + .

[0600] 1 H NMR(400MHz,Methanol-d4)δ8.47–8.34(m,2H),7.24-7.20(m,1H),7.10-7.14(m,1H ),6.79-6.68(m,1H),6.42–6.40(d,J=8.0Hz,1H),6.24–6.17(m,1H),5.75-5.71(m,1 H),5.01–4.94(m,2H),4.46-4.40(m,1H),4.26-4.17(m,1H),4.03-3.99(m,1H),3.84 -3.79(m,1H),2.86-2.77(m,1H),2.36(s,3H),1.26-1.19(m,9H),1.14-1.11(m,3H).

[0601] Example 13-1 and Example 13-2

[0602] (P-4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-7-(6-amino-3-chloro-2-fluorophenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one) and (M-4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-7-(6-amino-3-chloro-2-fluorophenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one)

[0603]

[0604] Example 13 was separated by SFC to obtain two axial chiral isomers, Example 13-1 and Example 13-2. SFC: Chiral preparation conditions:

[0605] Instrument: SFC-150 (Thar, Waters); column type: IC 20*250 mm, 10 μm (Daicel); column pressure: 100 bar; mobile phase: CO2 / Methanol (0.2% Methanol Ammonia) = 40 / 60; flow rate: 120 g / min; detection wavelength: UV 214 nm; column temperature: 35°C

[0606] Example 13-1:

[0607] t R =1.74min

[0608] MS m / z(ESI):656.1[M+H] + ,658.1[M+H+2] + .

[0609] 1 H NMR(400MHz,MeOD-d4)δ8.47–8.34(m,2H),7.24-7.20(m,1H),7.10-7.14(m,1H),6 .79-6.68(m,1H),6.42–6.40(d,J=8.0Hz,1H),6.24–6.17(m,1H),5.75-5.71(m,1H) ,5.01–4.94(m,2H),4.46-4.40(m,1H),4.26-4.17(m,1H),4.03-3.99(m,1H),3.84- 3.79(m,1H),2.86-2.77(m,1H),2.36(s,3H),1.26-1.19(m,9H),1.14-1.11(m,3H).

[0610] Example 13-2:

[0611] t R =2.49min

[0612] MS m / z(ESI):656.1[M+H] + ,658.1[M+H+2] + .

[0613] 1 H NMR(400MHz,DMSO-d6)δ8.55–8.38(m,2H),7.25-7.20(m,1H),7.18-7.11(m, 1H),6.88-6.76(m,1H),6.51–6.47(d,J=8.0Hz,1H),6.33–6.27(m,1H),5.84-5.80(m,1H),5.12–5.10(m,2H), 4.46-4.23(m,2H),4.15-3.89(m,2H),3.64-3.50(m,1H),2.89-2.82(m,1H),2.43(s,3H),1.51-0.99(m,12H).

[0614] Example 14

[0615] Preparation of 4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-6-chloro-7-(2-fluoro-6-hydroxyphenyl)-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one

[0616]

[0617] Preparation of 4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-6-chloro-7-(2-fluoro-6-hydroxyphenyl)-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one Reference Example 13.

[0618] MS m / z(ESI):623.1[M+H] + ,625.1[M+H+2] + .

[0619] 1H NMR(400MHz, Methanol-d4)δ8.47–8.34(m,2H),7.21-7.20(m,2H),6.89-6.77(m,1H),6. 64–6.55(m,2H),6.32–6.26(m,1H),5.84-5.80(m,1H),5.08–5.03(m,2H),4.56-4.49(m,1 H),4.34-4.26(m,1H),4.13-4.04(m,1H),3.92-3.88(m,1H),2.79-2.72(m,1H),2.40(s,3 H),1.55–1.43(m,3H),1.35-1.27(m,3H),1.20-1.17(m,3H),1.08-1.05(t,J=8.0Hz,3H).

[0620] Example 14-1 and Example 14-2

[0621] (P-4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-6-chloro-7-(2-fluoro-6-hydroxyphenyl)-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one) and (M-4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-6-chloro-7-(2-fluoro-6-hydroxyphenyl)-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one)

[0622]

[0623]

[0624] Example 14 was separated by SFC to obtain two axial chiral isomers, Example 14-1 and Example 14-2. SFC: Chiral preparation conditions:

[0625] Instrument: SFC-150 (Thar, Waters); column type: IC 20*250 mm, 10 μm (Daicel); column pressure: 100 bar; mobile phase: CO2 / Methanol (0.2% Methanol Ammonia) = 50 / 50; flow rate: 120 g / min; detection wavelength: UV 214 nm; column temperature: 35°C

[0626] Example 14-1:

[0627] t R =2.46min

[0628] MS m / z(ESI):623.1[M+H] + ,625.1[M+H+2] + .

[0629] 1 H NMR(400MHz, Methanol-d4)δ8.47–8.34(m,2H),7.21-7.20(m,2H),6.89-6.77(m,1H),6. 64–6.55(m,2H),6.32–6.26(m,1H),5.84-5.80(m,1H),5.08–5.03(m,2H),4.56-4.49(m,1 H),4.34-4.26(m,1H),4.13-4.04(m,1H),3.92-3.88(m,1H),2.79-2.72(m,1H),2.40(s,3 H),1.55–1.43(m,3H),1.35-1.27(m,3H),1.20-1.17(m,3H),1.08-1.05(t,J=8.0Hz,3H).

[0630] Example 14-2:

[0631] t R =3.08min

[0632] MS m / z(ESI):623.1[M+H] + ,625.1[M+H+2] + .

[0633] 1 H NMR(400MHz, Methanol-d4)δ8.48–8.34(m,2H),7.23-7.21(m,2H),6.90-6.78(m,1H),6. 66–6.58(m,2H),6.33–6.28(m,1H),5.85-5.82(m,1H),5.10–5.06(m,2H),4.58-4.50(m,1 H),4.34-4.27(m,1H),4.13-4.06(m,1H),3.93-3.88(m,1H),2.79-2.71(m,1H),2.41(s,3 H),1.56–1.46(m,3H),1.37-1.29(m,3H),1.21-1.18(m,3H),1.07-1.05(t,J=8.0Hz,3H).

[0634] Example 15

[0635] Preparation of (S)-4-(4-acryloyl-2-methylpiperazin-1-yl)-7-(2,6-difluorophenyl)-6-fluoro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one

[0636]

[0637] Preparation of (S)-4-(4-acryloyl-2-methylpiperazin-1-yl)-7-(2,6-difluorophenyl)-6-fluoro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one Reference Example 2.

[0638] MS m / z(ESI):595.1[M+H] + .

[0639] 1 H NMR(400MHz, Methanol-d4)δ8.40–8.32(m,2H),7.51(t,J=7.6Hz,1H),7.22(d,J=5.4Hz, 1H),7.05(t,J=8.4Hz,2H),6.86–6.79(m,1H),6.37–6.26(m,1H),5.84(d,J=10.6Hz,1H) ,5.08(m,2H),4.56-4.46(m,2H),4.21-4.08(m,1H),3.85-3.62(m,2H),2.86-2.82(m,1H ), 2.40 (s, 3H), 1.47 (d, J = 6.6Hz, 3H), 1.21–1.19 (d, J = 6.8Hz, 3H), 1.04 (d, J = 6.8Hz, 3H).

[0640] Example 16

[0641] Preparation of (S)-4-(4-acryloyl-2-methylpiperazin-1-yl)-6-fluoro-7-(2-fluorophenyl)-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one

[0642]

[0643] Preparation of (S)-4-(4-acryloyl-2-methylpiperazin-1-yl)-6-fluoro-7-(2-fluorophenyl)-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one Reference Example 2.

[0644] MS m / z(ESI):576.7[M+H]+

[0645] Example 17

[0646] Preparation of 4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-7-(2-amino-3,5-dichloro-6-fluorophenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one

[0647]

[0648] Preparation of 4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-7-(2-amino-3,5-dichloro-6-fluorophenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one Reference Example 13.

[0649] MS m / z(ESI):690.1[M+H] + ,692.1[M+H+2] + .

[0650] 1 H NMR(400MHz,Methanol-d4)δ8.46–8.34(m,2H),7.25-7.21(m,1H),7.11-7.14( m,1H),6.44–6.42(d,J=8.0Hz,1H),6.23–6.16(m,1H),5.73-5.70(m,1H),5.03– 4.97(m,2H),4.47-4.42(m,1H),4.25-4.16(m,1H),4.06-4.02(m,1H),3.86-3.8 3(m,1H),2.84-2.79(m,1H),2.34(s,3H),1.27-1.19(m,9H),1.16-1.14(m,3H).

[0651] Example 18

[0652] Preparation of 4-((S)-4-acryloyl-2-methylpiperazin-1-yl)-7-(2-amino-5-chloro-3,6-difluorophenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one

[0653]

[0654] Preparation of 4-((S)-4-acryloyl-2-methylpiperazin-1-yl)-7-(2-amino-5-chloro-3,6-difluorophenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one Reference Example 13.

[0655] MS m / z(ESI):660.1[M+H] + ,662.1[M+H+2] + .

[0656] 1 H NMR(400MHz, Methanol-d4)δ8.58–8.38(m,2H),7.53–7.36(m,1H),7.23–7.15(m,1H) ,6.97–6.79(m,1H),6.22(d,J=16Hz,1H),5.77(d,J=8Hz,1H),5.45–5.40(m,2H),5.0 7–4.82(m,1H),4.50–3.98(m,3H),3.92–3.49(m,2H),3.17–3.02(m,1H),2.93–2.63( m,1H),2.44–2.26(m,3H),1.43–1.27(m,3H),1.08(d,J=4Hz,3H),1.04–0.86(m,3H).

[0657] Example 19

[0658] Preparation of 4-((S)-4-acryloyl-2-methylpiperazin-1-yl)-7-(2-amino-5,6-difluoro-3-methylphenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one

[0659]

[0660] Preparation of 4-((S)-4-acryloyl-2-methylpiperazin-1-yl)-7-(2-amino-5,6-difluoro-3-methylphenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one Reference Example 13.

[0661] MS m / z(ESI):640.1[M+H] + ,642.1[M+H+2] + .

[0662] 1H NMR(400MHz, Methanol-d4)δ8.57–8.35(m,3H),7.25–7.04(m,2H),6.96–6.79( m,1H),6.29–6.14(m,1H),5.77(d,J=12Hz,1H),5.09–4.82(m,1H),4.76–4.58(m ,2H),4.48–3.98(m,3H),3.94–3.59(m,2H),2.93–2.69(m,1H),2.44–2.29(m,3H ),2.10–1.95(m,3H),1.42–1.26(m,3H),1.08(d,J=4Hz,3H),1.05–0.87(m,3H).

[0663] Example 20

[0664] Preparation of 4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-7-(2-amino-5-chloro-3,6-difluorophenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one

[0665]

[0666] Preparation of 4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-7-(2-amino-5-chloro-3,6-difluorophenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one Reference Example 13.

[0667] MS m / z(ESI):674.1[M+H] + ,676.1[M+H+2] + .

[0668] 1H NMR(400MHz, Methanol-d4)δ8.61–8.39(m,2H),7.56–7.35(m,1H),7.27–7.14( m,1H),6.96–6.75(m,1H),6.20(d,J=16Hz,1H),5.82–5.71(m,1H),5.53–5.38(m ,2H),4.95–4.69(m,1H),4.57–4.30(m,1H),4.24–4.00(m,2H),3.98–3.79(m,2 H),2.95–2.60(m,1H),2.44–2.25(m,3H),1.40–1.13(m,6H),1.10–0.87(m,6H).

[0669] Example 21

[0670] 4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-7-(2-amino-3,6-difluorophenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one

[0671]

[0672] Preparation of 4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-7-(2-amino-3,6-difluorophenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one Reference Example 13.

[0673] MS m / z(ESI):674.1[M+H] + ,

[0674] Example 22

[0675] Preparation of 4-((S)-4-acryloyl-2-methylpiperazin-1-yl)-7-(2-amino-3,6-difluorophenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one

[0676]

[0677] Preparation of 4-((S)-4-acryloyl-2-methylpiperazin-1-yl)-7-(2-amino-3,6-difluorophenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one Reference Example 13.

[0678] MS m / z(ESI):626.1[M+H] + ,628.1[M+H+2] + .

[0679] 1 H NMR(400MHz, Methanol-d4)δ8.58–8.34(m,2H),7.26–6.99(m,2H),6.95–6.77(m,1 H),6.47–6.27(m,1H),6.26–6.13(m,1H),5.77(d,J=16Hz,1H),5.22(s,2H),5.09–4 .80(m,1H),4.50–3.99(m,3H),3.95–3.53(m,2H),3.20–2.98(m,1H),2.94–2.65(m ,1H),2.42–2.24(m,3H),1.43–1.25(m,3H),1.09(d,J=4Hz,3H),1.04–0.82(m,3H).

[0680] Example 23

[0681] Preparation of 4-((S)-4-acryloyl-2-methylpiperazin-1-yl)-7-(2-amino-3,5,6-trifluorophenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one

[0682]

[0683] Refer to Example 13 for the preparation of 4-((S)-4-acryloyl-2-methylpiperazin-1-yl)-7-(2-amino-3,5,6-trifluorophenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one.

[0684] MS m / z(ESI):644.1[M+H]+.

[0685] Example 24

[0686] Preparation of 4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-7-(2-amino-3,5,6-trifluorophenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one

[0687]

[0688] Preparation of 4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-7-(2-amino-3,5,6-trifluorophenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one Reference Example 13.

[0689] MS m / z(ESI):658.1[M+H]+.

[0690] Example 25

[0691] Preparation of 4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-7-(2-amino-5,6-difluoro-3-methylphenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one

[0692]

[0693] Preparation of 4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-7-(2-amino-5,6-difluoro-3-methylphenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one Reference Example 13.

[0694] MS m / z(ESI):654.1[M+H] + ,656.1[M+H+2] + .

[0695] 1 H NMR(400MHz, Methanol-d4)δ8.58–8.31(m,2H),7.25–7.03(m,2H),6.94–6.73(m,1H),6.19(d,J=16Hz,1H),5.81–5.69(m,1H),4.96–4.59(m,3H),4.5 5–4.38(m,1H),4.29–3.96(m,2H),3.93–3.72(m,2H),3.00–2.60(m,1H),2 .45–2.25(m,3H),2.07–1.94(m,3H),1.43–1.13(m,6H),1.12–0.82(m,6H).

[0696] Example 26

[0697] Preparation of 4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-7-(6-amino-2,3,4-trifluorophenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one

[0698]

[0699] Preparation of 4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-7-(6-amino-2,3,4-trifluorophenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one Reference Example 13.

[0700] MS m / z(ESI):658.1[M+H]+,

[0701] Example 27

[0702] Preparation of 4-((S)-4-acryloyl-2-methylpiperazin-1-yl)-7-(6-amino-2,3,4-trifluorophenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one

[0703]

[0704] Preparation of 4-((S)-4-acryloyl-2-methylpiperazin-1-yl)-7-(6-amino-2,3,4-trifluorophenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one Reference Example 13.

[0705] MS m / z(ESI):644.1[M+H]+,

[0706] Example 28

[0707] Preparation of 4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-7-(6-amino-2,3-difluorophenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one

[0708]

[0709] Preparation of 4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-7-(6-amino-2,3-difluorophenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one Reference Example 13.

[0710] MS m / z(ESI):640.2[M+H]+,

[0711] Example 29

[0712] Preparation of 4-((S)-4-acryloyl-2-methylpiperazin-1-yl)-7-(6-amino-2,3-difluorophenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one

[0713]

[0714] Reference Example 13 was used to prepare 4-((S)-4-acryloyl-2-methylpiperazin-1-yl)-7-(6-amino-2,3-difluorophenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one.

[0715] MS m / z(ESI):626.1[M+H]+,

[0716] Example 30

[0717] Preparation of 4-((S)-4-acryloyl-2-methylpiperazin-1-yl)-6-chloro-7-(2-fluoro-6-methylphenyl)-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one

[0718]

[0719] Preparation of 4-((S)-4-acryloyl-2-methylpiperazin-1-yl)-6-chloro-7-(2-fluoro-6-methylphenyl)-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one Reference Example 13.

[0720] MS m / z(ESI):607.1[M+H] + ,609.1[M+H+2] + .

[0721] 1H NMR(400MHz, Methanol-d4)δ8.57–8.34(m,2H),7.43–7.31(m,1H),7.18(d,J=4Hz,1H),7.15–7 .01(m,2H),6.95–6.78(m,1H),6.28–6.14(m,1H),5.77(d,J=12Hz,1H),5.07–4.86(m,1H),4.4 5–4.25(m,2H),4.22–3.98(m,1H),3.93–3.58(m,2H),3.21–3.02(m,1H),2.87–2.69(m,1H),2. 40–2.27(m,3H),1.98–1.85(m,3H),1.41–1.28(m,3H),1.08(d,J=8Hz,3H),1.02–0.79(m,3H).

[0722] Example 31

[0723] Preparation of 4-((2S,5R)-4-acryloyl-2-methylpiperazin-1-yl)-6-chloro-7-(2-fluoro-6-methylphenyl)-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one

[0724]

[0725] Preparation of 4-((2S,5R)-4-acryloyl-2-methylpiperazin-1-yl)-6-chloro-7-(2-fluoro-6-methylphenyl)-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one Reference Example 13.

[0726] MS m / z(ESI):621.2[M+H]+,

[0727] Example 32

[0728] Preparation of 4-((S)-4-acryloyl-2-methylpiperazin-1-yl)-6-chloro-7-(2-chloro-6-fluorophenyl)-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one

[0729]

[0730] Preparation of 4-((S)-4-acryloyl-2-methylpiperazin-1-yl)-6-chloro-7-(2-chloro-6-fluorophenyl)-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one Reference Example 13.

[0731] MS m / z(ESI):627.1[M+H] + ,629.1[M+H+2] + .

[0732] 1 H NMR(400MHz, Methanol-d4)δ8.56–8.30(m,2H),7.58-7.36(m,3H),7.19(s,1H),6.87(s,1H),6.24-6.19(d,J=20.0Hz,1H),5 .79-5.76(d,J=12.0Hz,1H),4.97(s,1H),4.32-4.04(m,3H),3.80-3.49(m,3H),2.72(s,1H),2.35(s,3H),1.34-0.91(m,9H).

[0733] Example 33

[0734] Preparation of 4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-6-chloro-7-(2-chloro-6-fluorophenyl)-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one

[0735]

[0736] Preparation of 4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-6-chloro-7-(2-chloro-6-fluorophenyl)-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one Reference Example 13.

[0737] MS m / z(ESI):641.6[M+H]+,

[0738] Example 34

[0739] Preparation of (S)-4-(4-acryloyl-2-methylpiperazin-1-yl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)-7-(o-phenylmethyl)pyrido[2,3-d]pyrimidin-2(1H)-one

[0740]

[0741] Preparation of (S)-4-(4-acryloyl-2-methylpiperazin-1-yl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)-7-(o-phenylmethyl)pyrido[2,3-d]pyrimidin-2(1H)-one Reference Example 13.

[0742] MS m / z(ESI):589.1[M+H] + ,591.1[M+H+2] + .

[0743] Example 35

[0744] Preparation of (S)-4-(4-acryloyl-2-methylpiperazin-1-yl)-6-chloro-7-(2-chlorophenyl)-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one

[0745]

[0746] Preparation of (S)-4-(4-acryloyl-2-methylpiperazin-1-yl)-6-chloro-7-(2-chlorophenyl)-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one Reference Example 13.

[0747] MS m / z(ESI):609.6[M+H]+,

[0748] Example 36

[0749] Preparation of (S)-4-(4-acryloyl-2-methylpiperazin-1-yl)-6-chloro-7-(2-fluoro-6-(trifluoromethyl)phenyl)-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one

[0750]

[0751] Preparation of (S)-4-(4-acryloyl-2-methylpiperazin-1-yl)-6-chloro-7-(2-fluoro-6-(trifluoromethyl)phenyl)-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one Reference Example 13.

[0752] MS m / z(ESI):661.1[M+H]+,

[0753] Example 37

[0754] Preparation of 4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)-7-(o-phenylmethyl)pyrido[2,3-d]pyrimidin-2(1H)-one

[0755]

[0756] Preparation of 4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)-7-(o-phenylmethyl)pyrido[2,3-d]pyrimidin-2(1H)-one Reference Example 13.

[0757] MS m / z(ESI):603.1[M+H] + ,605.1[M+H+2] + .

[0758] Example 38

[0759] Preparation of 4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-6-chloro-7-(2-chlorophenyl)-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one

[0760]

[0761] Preparation of 4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-6-chloro-7-(2-chlorophenyl)-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one Reference Example 13.

[0762] MS m / z(ESI):623.6[M+H]+,

[0763] Example 39

[0764] Preparation of 4-((2S,5R)-(4-acryloyl-2,5-dimethylpiperazin-1-yl)-6-chloro-7-(2-fluoro-6-(trifluoromethyl)phenyl)-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one

[0765]

[0766] Preparation of 4-((2S,5R)-(4-acryloyl-2,5-dimethylpiperazin-1-yl)-6-chloro-7-(2-fluoro-6-(trifluoromethyl)phenyl)-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one Reference Example 13.

[0767] MS m / z(ESI):675.1[M+H]+,

[0768] Example 40

[0769] Preparation of 4-((S)-4-acryloyl-2-methylpiperazin-1-yl)-7-(2-amino-3,5-dichloro-6-fluorophenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one

[0770]

[0771] Preparation of 4-((S)-4-acryloyl-2-methylpiperazin-1-yl)-7-(2-amino-3,5-dichloro-6-fluorophenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one Reference Example 13.

[0772] MS m / z(ESI):676.1[M+H] + ,678.1[M+H+2] + .

[0773] 1 H NMR(400MHz, Methanol-d4)δ8.40–8.32(m,2H),7.51(t,J=7.6Hz,1H),7.22(d,J=5.4Hz, 1H),7.05(t,J=8.4Hz,2H),6.86–6.79(m,1H),6.37–6.26(m,1H),5.84(d,J=10.6Hz,1H) ,5.08(m,2H),4.56-4.46(m,2H),4.21-4.08(m,1H),3.85-3.62(m,2H),2.86-2.82(m,1H ), 2.40 (s, 3H), 1.47 (d, J = 6.6Hz, 3H), 1.21–1.19 (d, J = 6.8Hz, 3H), 1.04 (d, J = 6.8Hz, 3H).

[0774] 2. Biological Test Evaluation of Compounds

[0775] The present invention is further described and explained below in conjunction with test examples, but these examples are not intended to limit the scope of the present invention.

[0776] Test Example 1: Determination of the inhibitory effect on NCI-H358 / Mia PaCa-2 cell proliferation activity

[0777] 1.1 Experimental Purpose:

[0778] The inhibitory effect of the example compounds on the proliferation activity of KRAS G12C mutant cell lines NCI-H358 and Mia PaCa-2 cells was determined.

[0779] 1.2. Experimental instruments and reagents:

[0780] 1.2.1 Instruments:

[0781] Microplate reader (BioTek Synergy H1)

[0782] Pipette (Eppendorf & Rainin)

[0783] 1.2.2 Reagents:

[0784] NCI-H358 was purchased from Nanjing Kebai Biotechnology Co., Ltd.;

[0785] Mia PaCa-2 was purchased from ATCC;

[0786] Cell Titer-Glo cells were purchased from Promega, catalog number G7573;

[0787] RPMI 1640 was purchased from Gibco, catalog number 22400089;

[0788] DMEM was purchased from Gibco, catalog number 11995065;

[0789] FBS was purchased from Gibco, catalog number 10091148;

[0790] PBS was purchased from Gibco, catalog number 10010023;

[0791] Pancreatin was purchased from Gibco, catalog number 25200056;

[0792] Cell culture plates were purchased from Corning, catalog number 3610.

[0793] 1.3 Experimental methods

[0794] When NCI-H358 or Mia PaCa-2 cells were cultured to an appropriate degree of confluence, the NCI-H358 or Mia PaCa-2 cells were collected and adjusted to an appropriate cell concentration using complete culture medium. The cell suspension was plated into a 96-well plate at 90 μL per well and placed in a 37°C, 5% CO2 incubator to adhere overnight. Compound solutions of varying concentrations were prepared using DMSO and culture medium. A solvent control was set up and the compound solution was added to a 96-well plate at 10 μL per well. The plates were placed in a 37°C, 5% CO2 incubator and cultured for 72 hours. CellTiter-Glo solution was then added and mixed evenly by vortexing. The plates were incubated in the dark for 10 minutes and read using a BioTek Synergy H1 microplate reader.

[0795] 1.4. Experimental data processing method:

[0796] The inhibition rate was calculated using the luminescence signal value, and the concentration and inhibition rate were fitted with a nonlinear regression curve using Graphpad Prism software to obtain the IC 50 value.

[0797] 1.5. Experimental results:

[0798] The experimental results are shown in Table 8. The IC values ​​of the example compounds for the inhibition of proliferation of NCI-H358 and Mia PaCa-2 cells are 50 value.

[0799] Table 8

[0800]

[0801]

[0802] Note: “NT” means not tested.

[0803] 1.6. Experimental Conclusions:

[0804] According to the data, the example compounds of the present invention have a good proliferation inhibitory effect on NCI-H358 and Mia PaCa-2 cells.

[0805] Test Example 2: Determination of the ability of the compounds of the present invention to enhance the stability (melting temperature) of KRAS G12C protein

[0806] 2.1. Experimental Purpose:

[0807] The ability of the test compound to enhance the stability of the KRAS G12C protein (the degree of increase in the protein melting temperature can represent the binding ability of the compound to the KRAS G12C protein).

[0808] 2.2. Experimental reagents and instruments:

[0809] 2.2.1 Experimental instruments:

[0810] Quantitative PCR instrument (Quantstudio6Flex) was purchased from Life Science;

[0811] Pipettes were purchased from Eppendorf or Rainin.

[0812] 2.2.2 Experimental reagents:

[0813] Protein Thermal Shift TM Dye Kit was purchased from Thermofisher, catalog number 4461146;

[0814] KRAS G12C protein was purchased from Beijing Sino Biological Technology Co., Ltd., catalog number 12259-H07E2;

[0815] HEPES, 1M Buffer Solution was purchased from Thermofisher, catalog number 15630080;

[0816] DTT was purchased from Sigma, product number 43816-50mL;

[0817] NaCl was purchased from Sinopharm Chemical Reagent Co., Ltd. with the catalog number 10019318.

[0818] 2.3 Experimental methods:

[0819] In this experiment, the thermal shift method was used to test the change in the melting temperature (Tm) of KRAS G12C protein before and after compound binding to characterize the ability of the compound to enhance the stability of KRAS G12C protein.

[0820] The specific experimental operations are as follows:

[0821] Prepare an assay buffer containing 20 μM HEPES (pH 7.5), 1 mM DTT, 5X SYPRO Orange, and 150 mM NaCl. Add human KRAS G12C protein to a final concentration of 5.37 μM. Aliquot the reaction mixture into 8 PCR tube strips, 19.5 μL per tube. Add 0.5 μL of test compound or DMSO, for a total reaction volume of 20 μL, to a final compound concentration of 10 μM. A 2.5% DMSO control was used as a vehicle control. After incubation for one hour at room temperature in the dark, place the PCR tubes in a thermal cycler and launch QuantStudio Software v1.3. Use the melt curve function to measure the melting temperature of KRAS G12C protein in the different treatment groups (heating from 25°C to 95°C at 0.03°C / s).

[0822] 2.4. Experimental data processing method:

[0823] The PCR instrument experimental data file was imported into the thermal shift software to obtain the melting temperature (Tm) of each treatment group, and the Tm of the DMSO solvent control group was subtracted to obtain the change value of the melting temperature (ΔTm).

[0824] 2.5. Experimental results:

[0825] According to the above scheme, the compounds of the present invention showed the ability to increase the protein melting temperature as shown in Table 9 in the experiment of improving the stability of KRAS G12C protein.

[0826] Table 9

[0827] Example No. Tm (°C) DMSOTm (°C) ΔTm (°C) Example 1 48.66 0.2 11.6 Example 2 48.75 7.2 8.5 Example 3 50.66 1.5 10.9 Example 4 49.56 1.2 11.7 Example 5 48.66 4.4 15.8 Example 9 46.86 0.2 13.4 Example 13 47.05 8.0 11.0

[0828] 2.6 Experimental Conclusions:

[0829] The above data show that the example compounds of the present invention have good binding ability with KRAS G12C protein.

[0830] Test Example 3: Inhibitory activity of the compounds of the present invention on p-ERK in Miapaca-2 cells

[0831] 3.1. Experimental Purpose:

[0832] The inhibitory activity of the example compounds on the level of phosphorylated ERK in KRAS G12C mutant Mia PaCa-2 cells was determined.

[0833] 3.2. Experimental instruments:

[0834] 3.2.1 Instruments:

[0835] Microplate reader (BioTek Synergy H1);

[0836] Pipette (Eppendorf & Rainin).

[0837] 3.2.2 Reagents:

[0838] Phosphorylated ERK1 / 2 (T202-Y204) LANCE Ultra Cellular Detection Kit was purchased from PerkinElmer, catalog number TRF4000M;

[0839] Cell culture plates were purchased from Corning, catalog number 3610;

[0840] White opaque OptiPlate TM -384 plate was purchased from PerkinElmer, catalog number 6007290.

[0841] 3.3 Experimental methods

[0842] When Mia PaCa-2 cells reached an appropriate confluence, they were harvested and the cell density was adjusted to 1 × 10 6 / mL, the cell suspension was plated in a 96-well plate, 50 μL per well, and placed in a 37°C, 5% CO2 incubator to adhere overnight. Different concentrations of compound solutions were prepared using DMSO and complete culture medium. As a vehicle control, 25 μL of compound solution was added to each well of the 96-well plate. The plate was incubated in a 37°C, 5% CO2 incubator for another 2 hours. The supernatant was discarded from the cell culture plate, 50 μL of lysis buffer was added to each well, and the cells were shaken and lysed at room temperature for 30 minutes. The cells were centrifuged at 1000 rpm for 1 minute, and 15 μL of supernatant was transferred to a 384-well plate. 5 μL of detection mixture (Eu-labeled anti-ERK1 / 2 (T202-Y204) Antibody with a final detection concentration of 0.5 nM and ULight labeled anti-ERK1 / 2 Antibody with a final detection concentration of 5 nM) was added to each well. The cells were centrifuged at 1000 rpm for 1 minute to mix well. The cells were reacted overnight at room temperature and detected using BioTek Synergy H1 reads the plate and uses a time-resolved fluorescence program to detect the signal values ​​at emission wavelengths of 620 nm and 665 nm.

[0843] 3.4. Experimental data processing method:

[0844] The ratio of the signal values ​​at the emission wavelengths of 665 nm and 620 nm was calculated, and the inhibition rate was calculated using the ratio. The concentration and inhibition rate were fitted with a nonlinear regression curve using Graphpad Prism software to obtain the IC 50 value.

[0845] 3.5 Experimental results

[0846] Table 10 IC of pERK inhibition in Mia PaCa-2 cells 50 value

[0847]

[0848] 3.6. Experimental Conclusions

[0849] The above data show that the compounds of the present invention have a good inhibitory effect on pERK in Mia PaCa-2 cells.

[0850] Test Example 4: Pharmacokinetic Determination in Mice

[0851] 4.1. Research Objectives:

[0852] Balb / c mice were used as test animals to study the pharmacokinetic behavior of the compound in mice (plasma) after oral administration.

[0853] 4.2. Experimental plan:

[0854] 4.2.1 Investigational Drugs:

[0855] The compounds of the present invention were prepared in-house;

[0856] 4.2.2 Experimental Animals:

[0857] Balb / c male mice were purchased from Shanghai Jiesijie Experimental Animal Co., Ltd. with animal production license number (SCXK (Shanghai) 2013-0006N0.311620400001794).

[0858] 4.2.3 Drug preparation:

[0859] Weigh 5g of hydroxyethyl cellulose (HEC, CMC-Na, viscosity: 800-1200 cps), dissolve it in 1000mL of purified water, and add 10g of Tween 80. Mix well to form a clear solution.

[0860] The example compounds were weighed and added to 4-mL glass bottles, 2.4 mL of the solution was added, and ultrasonication was performed for 10 minutes to obtain a colorless clear solution with a concentration of 1 mg / mL.

[0861] 4.2.4 Administration:

[0862] Male Balb / c mice were fasted overnight and administered orally at a dose of 10 mg / kg in a volume of 10 mL / kg.

[0863] 4.2.5 Sample collection:

[0864] Blood was collected before administration and at 0.083h, 0.25h, 0.5h, 1h, 2h, 4h, 6h, and 8h after administration. The blood was placed in EDTA-2K tubes and centrifuged at 6000rpm for 6min at 4°C to separate the plasma, which was then stored at -80°C. Food was consumed 4h after administration. 4.3 Experimental Results:

[0865] The final determination results obtained by LCMS / MS method are shown in Table 11

[0866] Table 11: Pharmacokinetic parameters of compounds in mice

[0867]

[0868] 4.4 Experimental Conclusions

[0869] The above data show that the compounds of the present invention have good pharmacokinetic parameters in mice.

[0870] Test Example 5: Tumor Inhibition Experiment on MiaPaca 2 Transplanted Tumor Model

[0871] 5.1 Experimental Purpose:

[0872] BALB / c nude mice were used as test animals, and the human pancreatic cancer cell MiaPaca 2 xenograft (CDX) model was used for in vivo efficacy experiments to evaluate the anti-tumor effects of the test compounds.

[0873] 5.2 Experimental instruments and reagents:

[0874] 5.2.1 Instruments:

[0875] Clean bench (BSC-1300II A2, Shanghai Boxun Industrial Co., Ltd. Medical Equipment Factory);

[0876] CO2 incubator (Thermo-311, Thermo);

[0877] Centrifuge (Centrifuge 5720R, Eppendorf);

[0878] Automatic cell counter (Countess II, Life Technologies);

[0879] pipette (10-20 μL, Eppendorf);

[0880] microscope (Ts 2, Nikon);

[0881] Vernier caliper (CD-6”AX, Mitutoyo, Japan);

[0882] Cell culture flasks (T25 / T75 / T225, Corning);

[0883] Constant temperature water sink (HWS12, Shanghai Yiheng Science).

[0884] 5.2.2 Reagents:

[0885] DMEM (11995-065, Gibco);

[0886] Fetal bovine serum (FBS) (10091-148, Gibco);

[0887] 0.25% trypsin (25200-056, Gibco);

[0888] Penicillin-streptomycin dual antibody (P / S) (SV30010, GE);

[0889] Phosphate buffered saline (PBS) (10010-023, Gibco);

[0890] Matrigel (356234, Corning);

[0891] Gln (25030-081, Gibco).

[0892] 5.3 Experimental operation:

[0893] MiaPaca 2 cells were taken out from the cell bank, and after recovery, DMEM medium (containing 10% FBS, 1% Glu, 1% P / S) was added and placed in a CO2 incubator for culture (incubator temperature was 37°C, CO2 concentration was 5%). After the cells covered 80-90% of the bottom of the culture flask, they were passaged. After passage, the cells were continued to be cultured in a CO2 incubator. This process was repeated until the cell number met the in vivo drug efficacy inoculation requirement. Cells in the logarithmic growth phase were collected and counted using an automatic cell counter. According to the counting results, the cells were resuspended with PBS and Matrigel (volume ratio of 1:1) to make a cell suspension (density 8×10 7 / mL) and placed in an ice box for use.

[0894] The animals used were BALB / c nude mice, female, 6-8 weeks old, weighing approximately 18-22 grams. The mice were kept in an environment free of special pathogens and in single ventilated cages with 5 mice per cage. All cages, bedding and water were disinfected before use, and all animals had free access to standard certified commercial laboratory diets. Before the start of the experiment, the nude mice were marked with disposable universal ear tags for large and small mice. Before inoculation, the skin of the inoculation site was disinfected with 75% medical alcohol. Each mouse was inoculated subcutaneously on the right back with 0.1 ml (containing 8*10 6 When the average tumor volume reaches 100-200 mm 3 The test compound was administered orally daily at a dose / frequency of 6 mg / kg QD x 3 weeks. The efficacy of each group at the end of the experiment is shown in Table 5.

[0895] 5.4 Data Processing:

[0896] Tumor volume (mm) was measured twice a week using a vernier caliper. 3 ), calculated as: V = 0.5 * D * d * d, where D and d are the major and minor diameters of the tumor, respectively. Antitumor efficacy was determined by dividing the mean tumor volume increase in compound-treated animals by the mean tumor volume increase in untreated animals. Tumor inhibition rate (TGI) was calculated as: TGI (%) = 1 - [(Vt - V0) of the drug-treated group / (Vt - V0) of the solvent control group] * 100%. All animals were euthanized after the experiment.

[0897] 5.5 Experimental results:

[0898] Table 12: Pharmacodynamic parameters of compounds in transplanted tumor mice

[0899]

[0900] 5.6 Experimental Conclusions:

[0901] The above data show that after 21 consecutive days of oral administration, the example compound of the present invention can significantly inhibit the growth of MiaPaca 2 nude mouse transplanted tumor at a daily dose of 6 mg / kg.

[0902] Test Example 6: In vivo pharmacodynamic study on human lung cancer NCI-H358 cell xenograft tumor model

[0903] 6.1 Experimental Purpose:

[0904] The in vivo efficacy of the compounds was evaluated in a human lung cancer NCI-H358 cell xenograft tumor model.

[0905] 6.2 Experimental instruments and reagents:

[0906] 6.2.1 Instruments:

[0907] 1) Biological safety cabinet (BSC-1300II A2, Shanghai Boxun Industrial Co., Ltd. Medical Equipment Factory);

[0908] 2) Clean bench (CJ-2F, Suzhou Fengshi Experimental Animal Equipment Co., Ltd.);

[0909] 3) CO2 incubator (Thermo-311, Thermo);

[0910] 4) Centrifuge (Centrifuge 5720R, Eppendorf);

[0911] 5) Automated cell counter (Countess II, Life Technologies);

[0912] 6) Vernier caliper (CD-6"AX, Mitutoyo, Japan);

[0913] 7) Cell culture flasks (T75 / T225, Corning);

[0914] 8) Electronic balance (CPA2202S, Sartorius);

[0915] 9) Electronic balance (BSA2202S-CW, Sartorius);

[0916] 10) Electronic balance (BS124S, Sartorius).

[0917] 6.2.2 Reagents:

[0918] 1) RPMI-1640 medium (22400-089, Gibco);

[0919] 2) DMEM medium (11995-065, Gibco);

[0920] 3) Fetal bovine serum (FBS) (10099-141C, Gibco);

[0921] 4) Phosphate buffered saline (PBS) (10010-023, Gibco);

[0922] 5) Tween 80 (30189828, Sinopharm Reagent);

[0923] 6) Sodium carboxymethyl cellulose (30036365, Sinopharm Reagent.)

[0924] 6.3 Experimental operation and data processing:

[0925] 6.3.1 Animals:

[0926] BALB / c nude mice, 6-8 weeks old, female, were purchased from Shanghai Xipu-Bikai Laboratory Animal Co., Ltd.

[0927] 6.3.2 Cell culture and cell suspension preparation

[0928] 1) A strain of MiaPaca-2 cells was removed from the cell bank and revived with DMEM medium (DMEM + 10% FBS). The revived cells were placed in a cell culture flask (labeled with the cell type, date, and name of the culturer, etc.) and cultured in a CO2 incubator (incubator temperature: 37°C, CO2 concentration: 5%) (NCI-H358 cells were revived using the same method as the MiaPaca-2 cells in Test Example 5, except that the culture medium was changed to RPMI-1640 medium).

[0929] 2) Subculture the cells every three to five days, and continue to culture them in a CO2 incubator. Repeat this process until the cell count meets the in vivo efficacy requirement.

[0930] 3) The cultured MiaPaca-2 cells were collected and counted using an automatic cell counter. Based on the counting results, the cells were resuspended in PBS and Matrigel (ratio 1:1) to prepare a cell suspension (cell density 5×10 7 / mL) and placed in an ice box for use (NCI-H358 cells do not need to be added with Matrigel and can be directly resuspended in PBS with a cell density of 1×10 8 / mL).

[0931] 6.3.3 Sample preparation:

[0932] 1) Solvent: solvent (0.5% CMC-Na + 1% Tween 80), storage condition: 4°C.

[0933] Weigh 0.5 g of CMC-Na and dissolve it in a certain volume of ddH2O. Then add 1.0 mL of Tween 80 and stir to mix. Finally, dilute to 100 mL.

[0934] 2) Preparation of the test compound (10 mg / kg):

[0935] 8.42 mg of AMG510 compound was weighed, 8.260 mL of solvent was added, and a homogeneous solution was obtained by ultrasonication, vortexing, and stirring.

[0936] 7.81 mg of the compound of Example 13-1 was weighed, 7.654 mL of solvent was added, and a homogeneous solution was obtained by ultrasonication, vortexing, and stirring.

[0937] 6.3.3 Cell seeding

[0938] 1) Before inoculation, label the nude mice with disposable ear tags for both rats and mice;

[0939] 2) Mix the cell suspension thoroughly during inoculation, draw out 0.1-1 mL of cell suspension with a 1 mL syringe, remove any bubbles, and place the syringe on an ice pack until ready to use.

[0940] 3) Secure the nude mouse with your left hand and disinfect the right side of the mouse's back near the right shoulder (inoculation site) with 75% alcohol. After 30 seconds, begin inoculation.

[0941] 4) Inoculate the test nude mice sequentially (0.1 mL of cell suspension per mouse);

[0942] 6.3.4 Tumor measurement, grouping, and drug administration in tumor-bearing mice:

[0943] 1) Based on tumor growth, measure the tumor on day 18 after inoculation and calculate the tumor size.

[0944] Tumor volume calculation: Tumor volume (mm 3 ) = length (mm) × width (mm) × width (mm) / 2

[0945] 2) Tumor-bearing mice were randomly divided into groups according to their weight and tumor size.

[0946] 3) According to the grouping results, the test drug was started to be administered (administration method: oral administration; administration dose: 10 mg / kg; administration volume: 10 mL / kg; administration frequency: once / day; administration cycle: 21 days; solvent: 0.5% CMC / 1% Tween 80).

[0947] 4) After the start of the test drug administration, the tumors were measured and weighed twice a week.

[0948] 5) Euthanize the animals after the experiment.

[0949] 6) Use software such as Excel to process data.

[0950] 6.4 Data Processing:

[0951] Calculation of compound tumor inhibition rate (TGI) (%): When the tumor did not regress, TGI (%) = [(1-(average tumor volume at the end of treatment group dosing - average tumor volume at the start of treatment group dosing)) / (average tumor volume at the end of treatment in the solvent control group - average tumor volume at the start of treatment in the solvent control group)] × 100%. When the tumor regressed, TGI (%) = [1-(average tumor volume at the end of treatment group dosing - average tumor volume at the start of treatment group dosing) / average tumor volume at the start of treatment group dosing] × 100%.

[0952] 6.5 Experimental Results

[0953] Table 13: Pharmacodynamic parameters of compounds in transplanted tumor mice

[0954]

[0955] 6.6 Experimental Conclusions:

[0956] The above data show that after 15 consecutive days of oral administration, the example compound of the present invention can significantly inhibit the growth of human lung cancer NCI-H358 cell transplanted tumors in nude mice at a daily dose of 10 mg / kg, which is significantly better than the reference data.

[0957] Test Example 7: hERG potassium channel inhibitory activity test

[0958] 7.1 Cell Preparation

[0959] 7.1.1 CHO-hERG cells cultured at 175 cm 2 When the cell density in the culture flask grows to 60-80%, remove the culture medium, wash once with 7 mL PBS, and then add 3 mL Detachin for digestion.

[0960] 7.1.2 After digestion is complete, add 7 mL of culture medium to neutralize the cells, centrifuge, aspirate the supernatant, and resuspend in 5 mL of culture medium to ensure a cell density of 2 to 5 × 10 6 / mL.

[0961] 7.2 Solution preparation

[0962] Table 14: Components of intracellular and extracellular fluids

[0963]

[0964] 7.3 Electrophysiological Recording Process

[0965] The Qpatch instrument automatically performed the single-cell high-impedance sealing and whole-cell recording process. After acquiring the whole-cell recording mode, the cell was clamped at -80 mV. A 50-millisecond pre-voltage of -50 mV was applied before a 5-second depolarizing stimulus of +40 mV was applied. The cell then repolarized to -50 mV for 5 seconds before returning to -80 mV. This voltage stimulus was applied every 15 seconds. After recording for 2 minutes, extracellular solution was added for 5 minutes. Drug administration then began, starting with the lowest tested concentration. Each test concentration was administered for 2.5 minutes. After all concentrations were administered, the positive control compound, 3 μM Cisapride, was administered. At least three cells were tested for each concentration (n ≥ 3).

[0966] 7.4 Compound Preparation

[0967] 7.4.1 Dilute the 20 mM compound stock solution with extracellular fluid. Take 5 μL of the 20 mM compound stock solution and add it to 2495 μL of extracellular fluid to dilute it 500-fold to 40 μM. Then, perform three-fold serial dilutions in extracellular fluid containing 0.2% DMSO to obtain the final concentration to be tested.

[0968] 7.4.2 The highest tested concentration was 40 μM, and the other six concentrations were 40, 13.33, 4.44, 1.48, 0.49, and 0.16 μM, respectively.

[0969] 7.4.3 The DMSO content in the final test concentration does not exceed 0.2%. This concentration of DMSO has no effect on the hERG potassium channel.

[0970] 7.5 Data Analysis

[0971] The experimental data were analyzed by XLFit software.

[0972] 7.6 Quality Control

[0973] Environment: Humidity 20-50%, temperature 22-25°C

[0974] Reagents: All experimental reagents were purchased from Sigma, with a purity of >98%.

[0975] The experimental data in the report must meet the following standards:

[0976] Whole cell sealing impedance>100MΩ

[0977] Tail current amplitude>400pA

[0978] Pharmacological parameters:

[0979] The inhibitory effects of Cisapride at various concentrations on hERG channels were set as positive controls.

[0980] 7.7 Experimental results:

[0981] Table 15: Inhibition results of hERG current at multiple concentrations in the examples of the present invention

[0982] Example No. hERG (μM) Example 2-1>30 Example 9-1>30 Example 13-1>30 Example 14-1>30

[0983] 7.8 Experimental Conclusions:

[0984] The inhibition of cardiac hERG potassium ion channels by drugs is the main cause of drug-induced QT prolongation syndrome. From the experimental results, it can be seen that the compounds of the present invention have no significant inhibitory effect on cardiac hERG potassium ion channels, and can avoid cardiac toxicity and side effects at high doses.

[0985] Test Example 8: Plasma Stability Test Protocol

[0986] 8.1 Experimental Purpose

[0987] The purpose of this experiment is to test the stability of the example compounds in mouse, rat, dog and human plasma.

[0988] 8.2 Experimental Procedure

[0989] 8.2.1 Solution preparation

[0990] 1) Plasma preparation

[0991] After whole blood is collected from animals or humans, it is placed in a test tube containing an anticoagulant, centrifuged at 3500 rpm for 10 minutes, and the upper layer of light yellow plasma is collected.

[0992] 2) 10 μM test compound (m / M / V=C)

[0993] The compound was weighed, the stock solution was prepared with DMSO, and the working solution was prepared with 100 mM phosphate buffer.

[0994] 3) 10 μM positive control

[0995] (1) Propantheline (propantheline bromide Mr = 449.4 Da)

[0996] Weigh 2.36 mg of propantheline bromide and dilute it with 1 mL of DMSO to a 10 mM stock solution; pipette 10 μL of the 10 mM stock solution into 1 mL of 100 mM phosphate buffer for a final concentration of 100 μM.

[0997] (2) Mevinolin (lovastatin Mr = 404.5 Da)

[0998] Weigh 4.05 mg of lovastatin and dilute it with 1 mL of DMSO to a 10 mM stock solution; pipette 10 μL of the 10 mM stock solution into 1 mL of 100 mM phosphate buffer for a final concentration of 100 μM.

[0999] 8.2.2 Experimental process:

[1000] 1) In a 96-well plate, add 285 μL of plasma and 15 μL of 10 μM compound (test compound) in sequence and incubate at 37°C.

[1001] 2) Take out 40 μL at 0, 15, 30, 60, 90, and 120 min (the sampling point can be fine-tuned) and add 160 μL of acetonitrile stop solution containing internal standard.

[1002] 3) After centrifugation (3500 rpm, 10 min), 50 μL of supernatant was taken, diluted with 50 μL of DD H2O, and then injected into LC-MS / MS.

[1003] 8.3 Chromatographic conditions

[1004] Instrument: Shimadzu LC-20AD

[1005] Column: Phenomenex C18 (50*4.6mm, 5μm particle size);

[1006] Mobile phase: A: acetonitrile, B: 0.1% formic acid solution. 0-8 min: 5% A→95% A, 2.0-2.1 min: 90% A→5% A; flow rate: 0.8 mL / min; run time: 5.0 min; injection volume: 5 μL.

[1007] 8.4 Mass spectrometry conditions:

[1008] Instrument: API4000 liquid chromatography-mass spectrometer, AB Company, USA;

[1009] The ion source was electrospray ionization (ESI);

[1010] Drying gas (N2) temperature 500°C;

[1011] The electrospray voltage was 5500 V;

[1012] The detection method is positive ion detection;

[1013] The scanning mode was selected reaction monitoring (MRM);

[1014] The scan time is 0.1s.

[1015] 8.5 Experimental results:

[1016] Table 16: Plasma stability results of example compounds

[1017]

[1018]

[1019] 8.6 Experimental Conclusions:

[1020] The above data show that the compounds of the present invention have high plasma stability and small species differences.

[1021] Test Example 9: CYP enzyme single-point inhibition test

[1022] 9.1 Experimental Purpose

[1023] A human liver microsome incubation system was used to rapidly predict the inhibition of compounds on CYP450 enzyme isoforms using a single-point method.

[1024] 9.2 Experimental Procedure

[1025] 9.2.1 Solution preparation

[1026] For 2.5mM NADPH, weigh 4.165mg NADPH (reduced nicotinamide adenine dinucleotide phosphate) and add 100mM phosphate buffer to 2mL. For 0.25mg / mL microsomes, add 50μL of 20mg / mL microsomes to 4mL of 100mM phosphate buffer and mix thoroughly.

[1027] Preparation of test compound reaction solution

[1028] The compound of the example to be tested was weighed, diluted to 10 mM with DMSO, and then diluted to 100 μM with 100 mM phosphate buffer.

[1029] 9.2.2 Experimental process:

[1030] 1. In a 96-well plate, add 40 μL liver microsomes, 10 μL substrate, and 10 μL test compound and pre-incubate for 3 minutes.

[1031] 2. Add 40 μL of NADPH.

[1032] 3. At 20 min, add 300 μL of acetonitrile stop solution containing internal standard.

[1033] 4. Centrifuge and inject.

[1034] 9.3 Experimental Results

[1035] Table 17: Single-site inhibition results of CYP enzymes by example compounds

[1036]

[1037] Note:

[1038] Strong inhibition: IC 50 <1μM; moderate inhibition: 1μM <IC 50 <10 μM; weak inhibition: IC 50 >10μM

[1039] 9.4 Experimental Conclusions:

[1040] The above data show that the compounds of the present invention do not strongly inhibit various CYP enzyme subtypes and have a low DDI risk.

[1041] Test Example 10: Plasma protein binding rate experiment

[1042] 10.1 Experimental Purpose:

[1043] The purpose of this experimental method is to detect the plasma protein binding of the example compounds in plasma.

[1044] 10.2 Experimental instruments and materials:

[1045] Liquid chromatography-mass spectrometry instrument, centrifuge, vortexer, pipette, continuous liquid feeder, 96-well plate, tissue homogenizer (for tissue sample analysis), 50% methanol-water solution, acetonitrile solution with internal standard, blank matrix (plasma, urine, or tissue homogenate, etc.)

[1046] 10.3 Experimental steps:

[1047] 10.3.1 Preparation of Analyte Stock Solution A

[1048] The example compounds were prepared into 1 mM solution A using DMSO.

[1049] 10.3.2 Preparation of Plasma Solution B

[1050] Solution A was added to the plasma solution to prepare 5uM solution B.

[1051] 10.3.3 Processing Flow

[1052] 1) Add 200uL of solution B to the membrane.

[1053] 2) Add 350uL PBS to the outside of the membrane.

[1054] 3) Incubate in a 37°C water bath for 6 h.

[1055] 4) The sample is processed, diluted and then tested by mass spectrometry.

[1056] 10.4 Chromatographic conditions:

[1057] Instrument: Shimadzu LC-20AD;

[1058] Column: Phenomenex C18 (50*4.6mm, 5μm particle size);

[1059] Mobile phase: A: acetonitrile, B: 0.1% formic acid solution 0-0.5 min: 5% A→90% A, 2.0-2.1 min: 90% A→5% A; flow rate: 0.8 mL / min; run time: 5.0 min; injection volume: 5 μL.

[1060] 10.5 Mass spectrometry conditions:

[1061] Instrument: API4000 liquid chromatography-mass spectrometer, AB Company, USA;

[1062] The ion source was electrospray ionization (ESI);

[1063] Drying gas (N2) temperature 500°C;

[1064] The electrospray voltage was 5500 V;

[1065] The detection method is positive ion detection;

[1066] The scanning mode was selected reaction monitoring (MRM) and the scanning time was 0.1 s.

[1067] 10.6 Experimental Results:

[1068] Table 18: Plasma protein binding rate of example compounds

[1069] No. Human Rat Mouse Dog Example 2-198.090.588.482.6

[1070] Example 9 - 199.894.990.198.7 Example 13 - 199.797.993.998.7 Example 14 - 196.895.496.392.5

[1071] 10.7 Experimental Conclusions:

[1072] The above data show that the compounds of the present invention have high plasma protein binding rates with little species difference.

[1073] Test Example 11: Pharmacokinetic Determination in Tumor-Bearing Mice

[1074] 11.1. Research Objectives:

[1075] MiaPaca 2 tumor-bearing mice were used as test animals to study the pharmacokinetic behavior of Example 13-1 and AMG-510 compound at a dose of 6 mg / kg administered orally in mice (plasma, tumor tissue and intestine).

[1076] 11.2. Experimental Plan

[1077] 11.2.1 Investigational Drugs:

[1078] Example 13-1 of the present invention, AMG-510 compound, was prepared in-house.

[1079] 11.2.2 Experimental Animals:

[1080] Twenty-four female MiaPaca 2 tumor-bearing mice were used. Three mice were used at each time point (0 h, 1 h, 2 h, 4 h, 6 h, 8 h, 16 h, and 24 h). Shanghai Xipu-Bikai Laboratory Animal Co., Ltd., Animal Production License No. (SCXK(Shanghai)2018-0006).

[1081] 11.2.3 Drug preparation:

[1082] Weigh 5g of hydroxymethylcellulose, dissolve it in 1000mL of purified water, and add 10g of Tween 80. Mix well to form a clear solution.

[1083] Example compound 13-1 was weighed, AMG-510 was dissolved in the solution, shaken, and ultrasonicated for 15 minutes to obtain a uniform suspension with a concentration of 0.6 mg / mL.

[1084] 11.2.4 Administration:

[1085] MiaPaca 2 tumor-bearing mice were fasted and administered orally according to body weight (no drug was administered at 0 h) at a dose of 6 mg / kg in a volume of 10 mL / kg.

[1086] 11.2.5 Sample Collection:

[1087] Before and after dosing, mice were sacrificed by CO2, and 0.5 ml of cardiac blood was collected in EDTA-2K tubes. Plasma was separated by centrifugation at 6000 rpm for 6 min at 4°C and stored at -80°C. Tumor tissue was weighed, placed in 2 mL centrifuge tubes, and stored at -80°C. Appropriate lengths of duodenum, ileum, and colon tissue were cut open with scissors, the contents removed, and washed twice with PBS. The tissue was then blotted dry with absorbent paper, weighed, placed in 2 mL centrifuge tubes, and stored at -80°C.

[1088] 11.3 Experimental results: The final results were obtained by LCMS / MS method, see Table 11:

[1089] Table 19: Pharmacokinetic parameters of the compounds of the present invention in mice

[1090]

[1091]

[1092] 11.4 Experimental Conclusions:

[1093] At a dose of 6 mg / kg, the ratio of the exposure of the compound of the present invention in the mouse tumor to the exposure in the blood was higher than that of AMG-510. 1 / 2 And the MRT is longer.

[1094] 3. Study on the salts and crystal forms of the compounds

[1095] It is well known to those skilled in the art that when the compounds of the above examples are shown to have a good proliferation inhibitory effect on NCI-H358 and Mia PaCa-2 cells, their pharmaceutically acceptable salts often have the same pharmacological and pharmacodynamic activity. On this basis, the inventors further studied the physicochemical properties of the salt forms and crystal forms of the corresponding compounds. However, the preparation and characterization of the following specific salt forms or crystal forms do not limit the scope of protection of the present invention. Based on the present invention, those skilled in the art can obtain more salt forms and crystals of the compounds of the present invention through conventional salt formation or crystallization methods. These salt forms and crystals are all solutions protected by the present invention. Specifically as follows:

[1096] 1. Experimental instruments

[1097] 1.1 Some parameters of physical and chemical testing instruments

[1098]

[1099]

[1100] 1.2 Instruments and liquid analysis conditions

[1101] 1.2.1 Instruments and Equipment

[1102] Instrument Name and Model: METTLER TOLEDO XA105 analytical balance, Milli-Q Plus water purifier, Millipore Thermo Ultimate 3000 high-performance liquid chromatograph

[1103] 1.2.2 Chromatographic conditions

[1104]

[1105] 2. Research on salt forms of compounds

[1106] 2.1 Salt screening of compound of Example 13-1

[1107] 2.1.1 Experimental Purpose:

[1108] Screening of salt forms of compounds.

[1109] 2.1.2 Experimental steps:

[1110] 1) Instruments and equipment

[1111] Name Model Source Analytical Balance XA105 METTLER TOLEDO Ultrasonic Cleaner SK5200LHC Shanghai Kedao Ultrasonic Instrument

[1112] Eppendorf pipette (50mL, 100μL)

[1113] 2) Operating procedures

[1114] ①Dissolve or suspend into salt

[1115] Weigh 10 mg of the compound, add 200 μL of solvent, stir at room temperature, add different acids respectively, stir overnight, and centrifuge to dry or evaporate to obtain the salt of the compound.

[1116]

[1117] ② Anti-solvent method for salt formation

[1118] Select a good solvent, weigh the acid, add the good solvent to prepare a stock solution containing the compound at a concentration of 100 mg / mL, add the anti-solvent, weigh 100 mg of the compound, add 1 mL of the good solvent, and filter after all are dissolved. Take 0.2 mL of the filtrate and add the anti-solvent dropwise (stop adding if there is precipitation, and add a maximum of 1.8 mL of anti-solvent). After stirring for a period of time, quickly centrifuge to remove the filtrate to obtain the salt of the compound.

[1119]

[1120] 2.1.3 Experimental results:

[1121] Through salt screening experiments, sulfuric acid, isethionic acid, and 1,5-naphthalenedisulfonic acid were found to be capable of forming salts with the free base of the compound.

[1122] As described above, those skilled in the art can obtain more pharmaceutically acceptable salts by using conventional methods based on the present invention.

[1123] 2.2 Quantitative Analysis of Isethionate Salt of Compound 13-1

[1124] 2.2.1 HPLC quantification of isethionate

[1125] 2.2.1.1 Experimental Purpose:

[1126] Determine the number of isethionic acids in the isethionate salt of the compound of Example 13-1.

[1127] 2.2.1.2 Experimental steps:

[1128] 1) Chromatographic conditions

[1129]

[1130] 2) Operation

[1131] An appropriate amount of the free base of Example 13-1 was weighed and added with methanol to prepare a series of linear solutions with a concentration ranging from 0.05 to 0.30 mg / mL.

[1132] An appropriate amount of the isethionate salt compound of Example 13-1 was weighed and methanol was added to prepare a solution containing the isethionate salt of Example 13-1 at a concentration of 0.25 mg / mL. The linearization solution and the sample solution were injected separately.

[1133] 2.2.1.3 Experimental results:

[1134]

[1135]

[1136] The results of external standard method calculation showed that the isethionic acid and the free base formed salts in a 1:1 molar ratio.

[1137] 2.2.2 ELSD Quantification of Isethionate Salt of Compound 13-1

[1138] 2.2.2.1 Experimental Purpose:

[1139] Determine the number of isethionic acids in the isethionate salt of the compound of Example 13-1.

[1140] 2.2.2.2 Experimental steps:

[1141] 1) Chromatographic conditions

[1142] Dilutent MeOH Column ZIC-HILIC (150*4.6mm, 5μm) Mobile phase 75mM ammonium acetate solution (pH 4.80): acetonitrile = 30:70 Injection volume 5μL Flow rate 1.0mL / min Column Temperature 35℃ ELSD Temperature 40℃

[1143] 2) Operation

[1144] Weigh an appropriate amount of isethionic acid and add methanol to prepare a series of linear solutions containing isethionic acid in the range of 0.5-1 mg / mL.

[1145] An appropriate amount of the isethionate salt of the compound of Example 13-1 was weighed and methanol was added to prepare a solution containing the isethionate salt of the compound of Example 13-1 at a concentration of 5.0 mg / mL. The linearization solution and the sample solution were injected separately.

[1146] 2.2.2.3 Experimental results:

[1147]

[1148] The number of isethionic acid in the isethionic salt of the compound of Example 13-1 was calculated to be 1.

[1149] 3. Study on the crystal form of compound salts

[1150] 3.1 Study on the Crystalline Form of the Compound of Example 13-1

[1151] 3.1.1 Experimental Purpose:

[1152] Screening of the compound for crystalline salts.

[1153] 3.1.2 Experimental steps:

[1154] 1) Instruments and equipment

[1155]

[1156] 2) Operating procedures

[1157] ① Dissolve or suspend in different solvents to form salt crystals

[1158] 10 mg of the compound of Example 13-1 was weighed, and different reaction solvents were added respectively to make the final total volume 200 μL. The mixture was stirred, and acid was added. The mixture was stirred for 12 hours, centrifuged and dried, and then XRPD was measured.

[1159]

[1160]

[1161] ② Anti-solvent method for salt crystallization

[1162] Select a good solvent, weigh the acid, add the good solvent to prepare a stock solution containing the compound at a concentration of 100 mg / mL, add the anti-solvent, weigh 100 mg of the compound separately, add 1 mL of the good solvent, and filter after all are dissolved. Take 0.2 mL of the filtrate and dropwise add the anti-solvent (stop adding if there is precipitation, and add a maximum of 1.8 mL of anti-solvent). After stirring for a period of time, quickly centrifuge to remove the filtrate, and measure its XRPD after the solid is dried.

[1163]

[1164] 3.1.3 Experimental results

[1165] Through the crystal form research experiment of the salt of the compound, the crystal forms of the salts obtained are isethionate, sulfate, and 1,5-naphthalene disulfonate.

[1166] 3.2 Preparation of Crystalline Form of Compound 13-1

[1167] 3.2.1 Experimental Purpose:

[1168] Prepare the crystalline form of the salt of the compound of Example 13-1.

[1169] 3.2.2 Experimental steps:

[1170] 1) Instruments and equipment

[1171]

[1172] 2) Operating procedures

[1173] I. Preparation of Isethionate Crystalline Form I

[1174] 500 mg of the compound of Example 13-1 was weighed, 9.08 mL of isopropanol was added, and the mixture was heated at 50°C with stirring. 0.914 mL of isethionic acid (1.0 M in MeOH) was added, and the solution was dissolved to separate out a precipitate. The mixture was stirred at room temperature for 2 hours. After filtering, the solid was dried under vacuum at 50°C to obtain isethionic salt Form I. After testing and analysis, it had an XRPD pattern as shown in Figure 1, a DSC pattern as shown in Figure 2, and a TGA pattern as shown in Figure 3.

[1175] Alternatively, add 100 g of the compound of Example 13-1 and 1200 mL of isopropanol to a 3 L three-necked flask, heat to 40-45°C, and stir to dissolve. Then, disperse 2-hydroxyethylsulfonic acid (28.84 g) in 800 mL of ethanol and add dropwise to the reaction system, maintaining the temperature at 39-42°C over approximately 10 minutes. Add 500 mg of seed crystals to the reaction solution, causing a solid to precipitate rapidly. Remove from heat, cool to 25°C, and stir for 12 hours. Filter, wash the filter cake with 400 mL of isopropanol, drain, and vacuum dry at 45°C for 16 hours to yield 92.57 g of a pale yellow solid with a purity of 97.9%, a chiral purity of 99.8%, and a mass yield of 92%. Analysis and testing revealed an XRPD pattern substantially as shown in Figure 1, a DSC pattern substantially as shown in Figure 2, and a TGA pattern substantially as shown in Figure 3.

[1176] II. Preparation of Isethionate Crystalline Form II

[1177] 10 mg of the compound of Example 13-1 was weighed, 0.2 mL of tetrahydrofuran was added, and the mixture was heated and stirred at 50°C. 18.3 μL of isethionic acid (1.0 M in MeOH) was added, and the solution was dissolved to separate out a precipitate. The mixture was stirred at room temperature for 2 hours. After filtering, the solid was dried under vacuum at 50°C to obtain isethionic salt Form II. After testing and analysis, it had an XRPD pattern as shown in Figure 4, a DSC pattern as shown in Figure 5, and a TGA pattern as shown in Figure 6.

[1178] III. Preparation of Isethionate Crystalline Form III

[1179] 20 mg of the isethionate salt Form I was weighed, 0.2 mL of methanol and 0.45 mL of methyl tert-butyl ether were added, and the mixture was heated at 50° C. with stirring overnight. After filtration, the solid was dried under vacuum at 50° C. to obtain the isethionate salt Form III. After testing and analysis, the isethionate salt Form III had the XRPD pattern shown in FIG7 , the DSC pattern shown in FIG8 , and the TGA pattern shown in FIG9 .

[1180] IV. Preparation of Sulfate Crystal Form I

[1181] Weigh 10 mg of the compound of Example 13-1, add 0.2 mL of ethanol, heat and stir at 50°C, add 18.3 μL of sulfuric acid (1.0 M in MeOH), dissolve and precipitate, stir at room temperature overnight, filter the solid, and dry it in vacuo at 50°C to obtain sulfate salt Form I, which has an XRPD pattern as shown in Figure 10.

[1182] V. Preparation of Sulfate Crystal Form II

[1183] Weigh 100 mg of the compound of Example 13-1, add 1.82 mL of isopropanol, heat and stir at 50°C, add 183 μL of sulfuric acid (1.0 M in MeOH), dissolve and precipitate a solid, stir at room temperature overnight, filter the solid, and dry it in vacuo at 50°C to obtain sulfate salt Form II, which has an XRPD pattern as shown in Figure 11.

[1184] VI. Preparation of Sulfate Crystal Form III

[1185] 10 mg of sulfate salt form I was weighed, 0.2 mL of isopropanol was added, and the mixture was heated at 50° C. with stirring for 5 days. After filtration, the solid was dried under vacuum at 50° C. to obtain sulfate salt form III. After detection and analysis, it had an XRPD pattern as shown in FIG12 .

[1186] VII. Preparation of Sulfate Crystal Form IV

[1187] 10 mg of sulfate salt form I was weighed, 0.2 mL of ethyl acetate was added, and the mixture was heated and stirred at 50° C. for 5 days. After filtration, the solid was dried under vacuum at 50° C. to obtain sulfate salt form III. After detection and analysis, it had an XRPD pattern as shown in FIG13 .

[1188] 4. Solid Stability Test

[1189] 4.1 Immobilization Stability Experiment of Isethionate Crystal Form I of Compound 13-1

[1190] 4.1.1 Experimental Purpose:

[1191] The physical and chemical stability of the compound crystal form under high temperature, high humidity, high temperature and humidity and light conditions is investigated to provide a basis for crystal screening and storage.

[1192] 4.1.2 Instruments and liquid analysis conditions

[1193]

[1194] 4.1.3 Experimental plan

[1195] 4.1.3.1 An appropriate amount of the isethionate salt Form I of the compound of Example 13-1 was weighed and treated for a certain period of time under conditions of illumination (≥1.2×106 lux·h, 10 days), high humidity (25°C, 75%, 10 days), high humidity (25°C, 90%, 10 days), high temperature (40°C, 30 days), high temperature (60°C, 30 days), and micronization, and then its XRPD was measured.

[1196] 4.1.4.1 Experimental results:

[1197]

[1198] 4.1.3.2 Experimental plan:

[1199] An appropriate amount of the isethionate salt Form I of the compound of Example 13-1 was weighed and placed under light (5000±500 lux), high temperature (60°C), high humidity (92.5% RH), and high temperature and high humidity (50°C & 75% RH) conditions for 10 days. Then, methanol was added as a diluent to prepare a solution containing the free base of Example 13-1 at a concentration of 0.25 mg / mL. HPLC analysis was performed, and the changes in related substances were calculated according to the peak area normalization method.

[1200] 4.1.4.2 Experimental results:

[1201]

[1202] The above experimental results show that the isethionate salt form I of the compound in Example 13-1 is relatively stable under light, high humidity, high temperature and micro-powder conditions.

[1203] 4.2 Solid Stability Test of Compound Sulfate Form II in Example 13-1

[1204] 4.2.1 Experimental Purpose:

[1205] The physical and chemical stability of the compound crystal form under high temperature, high humidity, high temperature and humidity and light conditions is investigated to provide a basis for crystal screening and storage.

[1206] 4.2.2 Instruments and liquid analysis conditions

[1207]

[1208] 4.2.3 Experimental plan:

[1209] An appropriate amount of the sulfate salt form II of the compound of Example 13-1 was weighed and placed under light (5000±500 lux), high temperature (60°C), high humidity (92.5% RH), and high temperature and high humidity (50°C&75% RH) conditions for 10 days, respectively. Then, methanol as a diluent was added to prepare a solution containing the free base of Example 13-1 at a concentration of 0.25 mg / mL. HPLC analysis was performed, and the changes in related substances were calculated according to the peak area normalization method.

[1210] 4.2.4 Experimental results:

[1211]

[1212]

[1213] Sulfate crystal form II is relatively stable under light, high humidity, high temperature and high humidity conditions.

[1214] 5. Solubility experiments in different media

[1215] 5.1 Solubility test of compound 13-1 in different media

[1216] 5.1.1 Experimental Purpose:

[1217] The solubility of isethionate salt form I and sulfate salt form II in media with different pH values, water, artificial simulated gastric fluid (FaSSGF), fasting artificial simulated intestinal fluid (FaSSIF) and non-fasting artificial simulated intestinal fluid (FeSSIF) was investigated to provide a basis for the evaluation of the drugability of the salt.

[1218] 5.1.2 Experimental plan:

[1219] About 1 mg of different salt forms of the compound were weighed and suspended in 1 mL of artificial simulated gastric fluid (FaSSGF), fasting artificial simulated intestinal fluid (FaSSIF), non-fasting artificial simulated intestinal fluid (FeSSIF) and pure water for 24 hours. The thermodynamic solubility of the compound at 37°C was determined by HPLC and external standard method.

[1220] 5.1.3 Experimental results: as shown in the following table:

[1221]

[1222] 6. Thermodynamic stability experiments

[1223] 6.1 Polymorph Screening of the Isethionate Salt of the Compound of Example 13-1

[1224] 6.1.1 Experimental Purpose:

[1225] Through polycrystalline screening, a thermodynamically stable crystalline form of isethionate is obtained.

[1226] 6.1.2 Experimental plan:

[1227] 10 mg of isethionate salt Form I was taken, and 200 μL of organic solvent was added respectively. The mixture was slurried at room temperature and 50° C. for 5 days, centrifuged, and the supernatant was discarded. The solid was dried and its XRPD was measured.

[1228] 6.1.3 Experimental results: as shown in the following table:

[1229]

[1230]

[1231] The above results show that the isethionate salt crystal form I is a stable crystal form of isethionate salt.

[1232] 6.2 Polymorph Screening Experiment of Sulfate Salt of Compound 13-1

[1233] 6.2.1 Experimental Purpose:

[1234] Through polycrystalline screening, a thermodynamically stable sulfate crystal form is obtained.

[1235] 6.2.2 Experimental plan:

[1236] Take 10 mg of sulfate salt form II, add 200 μL of organic solvent respectively, beat at 50° C. for 5 days, centrifuge, discard the supernatant, and measure the XRPD of the solid after drying.

[1237] 6.2.3 Experimental results: as shown in the following table:

[1238] No. Solvent Sulfate - Initial Crystal Form Crystal Form II1 Ethanol Crystal Form II2 2-Methyltetrahydrofuran Crystal Form II3 2-Butanone Crystal Form II4 Ethyl acetate Crystal Form II5 Toluene Crystal Form II6 Isopropyl acetate Crystal Form II7 Tert-Butanol Crystal Form II

[1239] The above results show that sulfate crystal form II is a stable crystal form of sulfate.

Claims

1. Acid salts of compounds represented by general formula (II), in: R a is selected from hydrogen or methyl; R 1 is selected from hydrogen, fluorine, chlorine, bromine or methyl; R 3 Selected from hydrogen, amino, hydroxy, fluorine, chlorine, methyl, -S(CH 3 ) or trifluoromethyl; R 4 Selected from hydrogen, amino, hydroxy, fluorine, chlorine, -N(CH 3 ) 2 、-NH(CH 3 ) or fluorine; R 5 is selected from hydrogen, fluorine, chlorine, bromine, methyl, ethyl, propyl or isopropyl; R 6 is selected from hydrogen, fluorine, chlorine, bromine, methyl, ethyl, propyl or isopropyl; R 7 is selected from hydrogen, fluorine, chlorine, bromine or methyl; The acid of the acid salt is selected from hydrochloric acid, sulfuric acid, nitric acid, hydrobromic acid, hydrofluoric acid, hydroiodic acid, phosphoric acid, 2,5-dihydroxybenzoic acid, 1-hydroxy-2-naphthoic acid, acetic acid, ethanesulfonic acid, dichloroacetic acid, trichloroacetic acid, acetohydroxamic acid, adipic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, benzoic acid, 4-acetamidobenzoic acid, 4-aminobenzoic acid, capric acid, caproic acid, caprylic acid, cinnamic acid, citric acid, cyclohexanesulfamic acid, camphorsulfonic acid, aspartic acid, camphoric acid, gluconic acid, glucuronic acid, glutamic acid, isoascorbic acid, lactic acid, malic acid, mandelic acid, pyroglutamic acid, tartaric acid, dodecylsulfuric acid, dibenzoyltartaric acid, ethane-1,2-dihydrogen iodide sulfonic acid, ethanesulfonic acid, formic acid, fumaric acid, galactosonic acid, gentisic acid, glutaric acid, 2-ketoglutaric acid, glycolic acid, hippuric acid, isethionic acid, lactobionic acid, ascorbic acid, aspartic acid, lauric acid, camphoric acid, maleic acid, malonic acid, methanesulfonic acid, 1,5-naphthalenedisulfonic acid, naphthalene-2-sulfonic acid, nicotinic acid, oleic acid, orotic acid, oxalic acid, palmitic acid, pamoic acid, propionic acid, salicylic acid, 4-aminosalicylic acid, sebacic acid, stearic acid, succinic acid, thiocyanic acid, undecylenic acid, trifluoroacetic acid, benzenesulfonic acid, p-toluenesulfonic acid or L-malic acid; preferably hydrochloric acid, phosphoric acid, ethanesulfonic acid, benzenesulfonic acid, methanesulfonic acid, fumaric acid, isethionic acid, oxalic acid or hydrobromic acid.

2. The acid salt of the compound according to claim 1, It is characterized in that The compound is further represented by the general formula (II-A) or (II-B):

3. An acid salt of the compound according to claim 1 or 2, It is characterized in that The compound is selected from:

4. An acid salt of the compound according to any one of claims 1 to 3, It is characterized in that The compound is P-4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-7-(6-amino-3-chloro-2-fluorophenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one; P-4-((S)-4-acryloyl-2-methylpiperazin-1-yl)-6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one; P-4-((S)-4-acryloyl-2-methylpiperazin-1-yl)-7-(2-amino-6-fluorophenyl)-6-fluoro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one; P-4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-6-chloro-7-(2-fluoro-6-hydroxyphenyl)-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one; P-4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one; The acid in the acid salt is selected from isethionic acid, sulfuric acid, 1,5-naphthalene disulfonic acid, methanesulfonic acid, hydrobromic acid, phosphoric acid, benzenesulfonic acid, oxalic acid, maleic acid, adipic acid, hydrochloric acid, citric acid, malonic acid, L-malic acid, pamoic acid, p-toluenesulfonic acid or fumaric acid, preferably isethionic acid or sulfuric acid.

5. An acid salt of the compound according to any one of claims 1 to 4, It is characterized in that The number of acids is 0.2-3; preferably 0.2, 0.5, 1, 1.5, 2, 2.5 or 3; more preferably 0.5, 1, 2 or 3, further preferably 1.

6. An acid salt of the compound according to any one of claims 1 to 5, It is characterized in that The acid salt is a hydrate or an anhydrate; when the acid salt is a hydrate, the number of water is 0.2-3; preferably 0.2, 0.5, 1, 1.5, 2, 2.5 or 3; more preferably 0.5, 1, 2 or 3.

7. An acid salt of the compound according to any one of claims 1 to 6, It is characterized in that The acid salt is in crystalline form; The preferred compound is the acid salt crystalline form of P-4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-7-(6-amino-3-chloro-2-fluorophenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one; Acid salt form of P-4-((S)-4-acryloyl-2-methylpiperazin-1-yl)-6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one; Acid salt form of P-4-((S)-4-acryloyl-2-methylpiperazin-1-yl)-7-(2-amino-6-fluorophenyl)-6-fluoro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one; Acid salt form of P-4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-6-chloro-7-(2-fluoro-6-hydroxyphenyl)-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one; Acid salt form of P-4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one; More preferred are isethionate salt form, sulfate salt form, 1,5-naphthalene disulfonate salt form, methanesulfonate salt form, hydrobromide salt form, phosphate salt form, benzenesulfonate salt form, oxalate salt form, maleate salt form, adipate salt form, hydrochloride salt form, citrate salt form, malonate salt form, L-malate salt form, pamoate salt form, p-toluenesulfonate salt form or fumarate salt form.

8. The acid salt of the compound according to claim 7, Features: The acid salt crystalline form of P-4-((2S,5R)-4-acryloyl-2,5-dimethylpiperazine-1-yl)-7-(6-amino-3-chloro-2-fluorophenyl)-6-chloro-1-(2-isopropyl-4-(methylthio)pyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one is: Isethionate salt form I, whose X-ray powder diffraction spectrum 2θ has a diffraction peak at 21.7±0.2°; or has a diffraction peak at 8.8±0.2°; or has a diffraction peak at 19.3±0.2°; or has a diffraction peak at 27.6±0.2°; Or having a diffraction peak at 10.9±0.2°; or having a diffraction peak at 15.4±0.2°; or having a diffraction peak at 16.7±0.2°; or having a diffraction peak at 15.8±0.2°; or having a diffraction peak at 17.5±0.2°; or having a diffraction peak at 23.8±0.2°; or having a diffraction peak at 10.2±0.2°; or having a diffraction peak at 11.8±0.2°; preferably including any 2-5, or 3-5, or 3-6, or 3-8, or 5-8, or 6-8 of the above diffraction peaks, more preferably including any 6, 7 or 8 thereof; It is isethionate salt form II, and its X-ray powder diffraction spectrum 2θ has a diffraction peak at 21.7±0.2°; or a diffraction peak at 8.8±0.2°; or a diffraction peak at 19.3±0.2°; or a diffraction peak at 27.6±0.2°; or a diffraction peak at 10.9±0.2°; or a diffraction peak at 23.8±0.2°; or a diffraction peak at 16.7±0.2°; or a diffraction peak at 15.4±0.2°; or a diffraction peak at 15.8±0.2°; or a diffraction peak at 10.0±0.2°; preferably includes any 2-5, or 3-5, or 3-6, or 3-8, or 5-8, or 6-8 of the above diffraction peaks, and more preferably includes any 6, 7 or 8 thereof; It is a hydroxyethyl sulfonate crystal form III, and its X-ray powder diffraction spectrum 2θ has a diffraction peak at 19.4±0.2°; or a diffraction peak at 16.9±0.2°; or a diffraction peak at 26.6±0.2°; or a diffraction peak at 14.6±0.2°; or a diffraction peak at 28.0±0.2°; or a diffraction peak at 25.6±0.2°; or a diffraction peak at 2 A diffraction peak at 0.7±0.2°; or a diffraction peak at 12.8±0.2°; or a diffraction peak at 19.1±0.2°; or a diffraction peak at 27.2±0.2°; preferably including any 2-5, or 3-5, or 3-6, or 3-8, or 5-8, or 6-8 of the above diffraction peaks, more preferably including any 6, 7 or 8 thereof; It is sulfate crystal form I, and its X-ray powder diffraction spectrum 2θ has a diffraction peak at 19.0±0.2°; or a diffraction peak at 19.4±0.2°; or a diffraction peak at 12.4±0.2°; or a diffraction peak at 26.2±0.2°; or a diffraction peak at 17.6±0.2°; or a diffraction peak at 18.1±0.2°; or a diffraction peak at 25.3±0.2°; or a diffraction peak at 8.8±0.2°; or a diffraction peak at 21.9±0.2°; or a diffraction peak at 11.5±0.2°; preferably includes any 2-5, or 3-5, or 3-6, or 3-8, or 5-8, or 6-8 of the above diffraction peaks, and more preferably includes any 6, 7 or 8 thereof; It is sulfate crystal form II, and its X-ray powder diffraction spectrum 2θ has a diffraction peak at 15.5±0.2°; or a diffraction peak at 11.1±0.2°; or a diffraction peak at 8.9±0.2°; or a diffraction peak at 19.3±0.2°; or a diffraction peak at 22.3±0.2°; or a diffraction peak at 23.6±0.2°; or a diffraction peak at 17.4±0.2°; or a diffraction peak at 27.3±0.2°; or a diffraction peak at 17.0±0.2°; or a diffraction peak at 27.9±0.2°; preferably includes any 2-5, or 3-5, or 3-6, or 3-8, or 5-8, or 6-8 of the above diffraction peaks, and more preferably includes any 6, 7 or 8 thereof; It is sulfate crystal form III, and its X-ray powder diffraction spectrum 2θ has a diffraction peak at 19.6±0.2°; or a diffraction peak at 18.0±0.2°; or a diffraction peak at 18.4±0.2°; or a diffraction peak at 16.8±0.2°; or a diffraction peak at 14.3±0.2°; or a diffraction peak at 11.8±0.2°; or a diffraction peak at 14.9±0.2°; or a diffraction peak at 25.7±0.2°; or a diffraction peak at 15.4±0.2°; or a diffraction peak at 23.5±0.2°; preferably includes any 2-5, or 3-5, or 3-6, or 3-8, or 5-8, or 6-8 of the above diffraction peaks, and more preferably includes any 6, 7 or 8 thereof; It is sulfate crystal form IV, and its X-ray powder diffraction spectrum has a diffraction peak at 19.4±0.2°; or a diffraction peak at 18.9±0.2°; or a diffraction peak at 15.5±0.2°; or a diffraction peak at 8.8±0.2°; or a diffraction peak at 18.1±0.2°; or a diffraction peak at 24.9±0.2°; or a diffraction peak at 17.4±0.2°; or a diffraction peak at 12.3±0.2°; or a diffraction peak at 26.1±0.2°; or a diffraction peak at 14.5±0.2°; preferably includes any 2-5, or 3-5, or 3-6, or 3-8, or 5-8, or 6-8 of the above diffraction peaks, and more preferably includes any 6, 7 or 8 of them.

9. The acid salt of the compound according to claim 8, Features: The X-ray powder diffraction pattern of the isethionate salt form I comprises at least one or more diffraction peaks located at 2θ of 21.7±0.2°, 8.8±0.2°, and 19.3±0.2°, preferably two of them, and more preferably three of them; optionally, it may further comprise at least one of 2θ of 27.6±0.2°, 10.9±0.2°, 15.4±0.2°, 16.7±0.2°, 15.8±0.2°, 10.2±0.2°, and 11.8±0.2°, preferably 2, 3, 4 or 5 of them; for example, 8.8±0.2°、27.6±0.2°； 21.7±0.2°、8.8±0.2°、10.9±0.2°； 21.7±0.2°、8.8±0.2°、27.6±0.2°、10.9±0.2°； 15.8±0.2°、8.8±0.2°、27.6±0.2°、10.9±0.2°；21.7±0.2°、8.8±0.2°、19.3±0.2°、15.8±0.2°、10.9±0.2°、15.4±0.2°； 10.9±0.2°、8.8±0.2°、10.2±0.2°、27.6±0.2°、10.9±0.2°、15.8±0.2°； The X-ray powder diffraction pattern of the isethionate salt form II contains at least one or more diffraction peaks located at 2θ of 21.7±0.2°, 10.0±0.2°, and 8.8±0.2°, preferably two of them, and more preferably three of them; optionally, it may further contain at least one of 2θ of 19.3±0.2°, 27.6±0.2°, 10.9±0.2°, 23.8±0.2°, and 16.7±0.2°, preferably 2, 3, 4 or 5 of them; for example, 21.7±0.2°、10.0±0.2°； 21.7±0.2°、10.0±0.2°、19.3±0.2°； 21.7±0.2°、10.0±0.2°、8.8±0.2°、19.3±0.2°； 21.7±0.2°、10.0±0.2°、8.8±0.2°、16.7±0.2°、27.6±0.2°、10.9±0.2°； The X-ray powder diffraction pattern of the isethionate salt form III comprises at least one or more diffraction peaks located at 2θ of 19.4±0.2°, 16.9±0.2°, and 26.6±0.2°, preferably two of them, and more preferably three of them; optionally, it may further comprise at least one of 2θ of 14.6±0.2°, 28.0±0.2°, 25.6±0.2°, 20.7±0.2°, and 12.8±0.2°, preferably 2, 3, 4 or 5 of them; for example, 19.4±0.2°、16.9±0.2°、26.6±0.2°、14.6±0.2°、28.0±0.2°、25.6±0.2°； The X-ray powder diffraction pattern of the sulfate salt form I comprises at least one or more diffraction peaks located at 2θ of 19.0±0.2°, 19.4±0.2°, and 12.4±0.2°, preferably two of them, and more preferably three of them; optionally, it may further comprise at least one of 2θ of 26.2±0.2°, 17.6±0.2°, 18.1±0.2°, 25.3±0.2°, and 8.8±0.2°, preferably 2, 3, 4 or 5 of them; for example, 19.0±0.2°、19.4±0.2°、12.4±0.2°、26.2±0.2°、17.6±0.2°、18.1±0.2°； The X-ray powder diffraction pattern of the sulfate crystal form II contains at least one or more diffraction peaks located at 2θ of 15.5±0.2°, 11.1±0.2°, and 8.9±0.2°, preferably two of them, and more preferably three of them; optionally, it may further contain at least one of 2θ of 19.3±0.2°, 22.3±0.2°, 23.6±0.2°, 17.4±0.2°, and 27.3±0.2°, preferably 2, 3, 4 or 5 of them; for example, 15.5±0.2°、11.1±0.2°； 15.5±0.2°、11.1±0.2°、8.9±0.2°； 15.5±0.2°、11.1±0.2°、8.9±0.2°、19.3±0.2°； 15.5±0.2°、11.1±0.2°、8.9±0.2°、19.3±0.2°、22.3±0.2°、27.3±0.2°； The X-ray powder diffraction pattern of the sulfate crystalline form III contains at least one or more diffraction peaks located at 2θ of 19.6±0.2°, 18.0±0.2°, and 18.4±0.2°, preferably two of them, and more preferably three of them; optionally, it may further contain at least one of 2θ of 16.8±0.2°, 14.3±0.2°, 11.8±0.2°, 14.9±0.2°, and 25.7±0.2°, preferably 2, 3, 4 or 5 of them; for example, 19.6±0.2°、18.0±0.2°、18.4±0.2°、16.8±0.2°、14.3±0.2°、11.8±0.2°； The X-ray powder diffraction pattern of sulfate crystal form IV contains at least one or more diffraction peaks located at 2θ of 19.4±0.2°, 18.9±0.2°, and 15.5±0.2°, preferably two of them, and more preferably three of them; optionally, it may further contain at least one of 2θ of 8.8±0.2°, 18.1±0.2°, 24.9±0.2°, 17.4±0.2°, and 12.3±0.2°, preferably 2, 3, 4 or 5 of them; for example, 19.4±0.2°、18.9±0.2°、15.5±0.2°、8.8±0.2°、18.1±0.2°、24.9±0.2°。 10. An acid salt of the compound according to claim 8 or 9, Features: The X-ray powder diffraction pattern of the isethionate salt form I optionally further comprises one or more diffraction peaks located at 2θ of 21.7±0.2°, 8.8±0.2°, 10.2±0.2°, 11.8±0.2°, 13.3±0.2°, 19.3±0.2°, 27.6±0.2°, 10.9±0.2°, 15.4±0.2°, 16.7±0.2°, 15.8±0.2°, 17.5±0.2°, 23.8±0.2°; preferably, at least any 2-3, or 4-5, or 6-8 thereof; further preferably, any 2, 3, 4, 5, 6, 7 or 8 thereof; for example, 8.8±0.2°、10.2±0.2°、11.8±0.2°、13.3±0.2°、27.6±0.2°、10.9±0.2°、15.8±0.2°、17.5±0.2°； 21.7±0.2°、8.8±0.2°、19.3±0.2°、27.6±0.2°、10.9±0.2°、17.5±0.2°、16.7±0.2°、15.8±0.2°； The X-ray powder diffraction pattern of the isethionate salt form II optionally further comprises one or more diffraction peaks located at 2θ of 10.0±0.2°, 21.7±0.2°, 8.8±0.2°, 19.3±0.2°, 27.6±0.2°, 10.9+±0.2°, 23.8±0.2°, 16.7±0.2°, 15.4±0.2°, 15.8±0.2°, 10.0±0.2°; preferably, at least any 2-3, or 4-5, or 6-8 thereof; further preferably, any 2, 3, 4, 5, 6, 7 or 8 thereof; for example, 21.7±0.2°、8.8±0.2°、19.3±0.2°、27.6±0.2°、10.9+±0.2°、23.8±0.2°、16.7±0.2°、15.4±0.2°； The X-ray powder diffraction pattern of the isethionate salt form III optionally further comprises one or more diffraction peaks located at 2θ of 19.4±0.2°, 16.9±0.2°, 26.6±0.2°, 14.6±0.2°, 28.0±0.2°, 25.6±0.2°, 20.7±0.2°, 12.8±0.2°, 19.1±0.2°, 27.2±0.2°; preferably, at least any 2-3, or 4-5, or 6-8 thereof; further preferably, any 2, 3, 4, 5, 6, 7 or 8 thereof; for example, 19.4±0.2°、16.9±0.2°、26.6±0.2°、14.6±0.2°、28.0±0.2°、25.6±0.2°、20.7±0.2°、27.2±0.2°； The X-ray powder diffraction pattern of the sulfate salt form I optionally further comprises one or more diffraction peaks located at 2θ of 19.0±0.2°, 19.4±0.2°, 12.4±0.2°, 26.2±0.2°, 17.6±0.2°, 18.1±0.2°, 25.3±0.2°, 8.8±0.2°, 21.9±0.2°, 11.5±0.2°; preferably, at least any 2-3, or 4-5, or 6-8 thereof; further preferably, any 2, 3, 4, 5, 6, 7 or 8 thereof; for example, 19.0±0.2°、19.4±0.2°、12.4±0.2°、26.2±0.2°、25.3±0.2°、8.8±0.2°、21.9±0.2°、11.5±0.2°； The X-ray powder diffraction pattern of the sulfate salt form II optionally further comprises one or more diffraction peaks located at 2θ of 15.5±0.2°, 11.1±0.2°, 8.9±0.2°, 19.3±0.2°, 22.3±0.2°, 23.6±0.2°, 17.4±0.2°, 27.3±0.2°, 17.0±0.2°, 27.9±0.2°; preferably, at least any 2-3, or 4-5, or 6-8 thereof; further preferably, any 2, 3, 4, 5, 6, 7 or 8 thereof; for example, 15.5±0.2°、8.9±0.2°、19.3±0.2°、22.3±0.2°、23.6±0.2°、17.4±0.2°、27.3±0.2°、17.0±0.2°； The X-ray powder diffraction pattern of sulfate salt form III optionally further comprises one or more diffraction peaks located at 2θ of 19.6±0.2°, 18.0±0.2°, 18.4±0.2°, 16.8±0.2°, 14.3±0.2°, 11.8±0.2°, 14.9±0.2°, 25.7±0.2°, 15.4±0.2°, 23.5±0.2°; preferably at least any 2-3, or 4-5, or 6-8 thereof; further preferably, any 2, 3, 4, 5, 6, 7 or 8 thereof; for example, 19.6±0.2°、18.0±0.2°、18.4±0.2°、16.8±0.2°、14.3±0.2°、11.8±0.2°、14.9±0.2°、23.5±0.2°； The X-ray powder diffraction pattern of sulfate salt form IV optionally further comprises one or more diffraction peaks located at 2θ of 19.4±0.2°, 18.9±0.2°, 15.5±0.2°, 8.8±0.2°, 18.1±0.2°, 24.9±0.2°, 17.4±0.2°, 12.3±0.2°, 26.1±0.2°, 14.5±0.2°; preferably at least any 2-3, or 4-5, or 6-8 thereof; further preferably, any 2, 3, 4, 5, 6, 7 or 8 thereof; for example, 19.4±0.2°、18.9±0.2°、15.5±0.2°、8.8±0.2°、18.1±0.2°、24.9±0.2°、17.4±0.2°、12.3±0.2°。 11. The acid salt of the compound according to claim 8, Features: The X-ray powder diffraction pattern of the isethionate salt form I comprises 2θ at 21.7±0.2°, 8.8±0.2°, 10.2±0.2°, 11.8±0.2°, 13.1±0.2°, 19.3±0.2°, 27.6±0.2°, 10.9±0.2°, 13.3±0.2°, 15.4±0.2°, 16.7±0.2° , 15.8±0.2°, 17.5±0.2°, 23.8±0.2°, 14.7±0.2°, 24.3±0.2°, 27.3±0.2°, 23.4±0.2°, 20.6±0.2°, 21.2±0.2°, one or more diffraction peaks, preferably, including 4, 5, 6, 8 or 10 diffraction peaks selected therefrom; for example, 8.8±0.2°、19.3±0.2°、27.6±0.2°、20.6±0.2°； 8.8±0.2°、19.3±0.2°、27.6±0.2°、10.9±0.2°、15.4±0.2°、20.6±0.2°； 21.7±0.2°、8.8±0.2°、19.3±0.2°、27.6±0.2°、10.9±0.2°、15.4±0.2°、16.7±0.2°、20.6±0.2°； 21.7±0.2°、8.8±0.2°、19.3±0.2°、27.6±0.2°、10.9±0.2°、15.4±0.2°、16.7±0.2°、15.8±0.2°、24.3±0.2°、23.8±0.2°； 8.8±0.2°、10.2±0.2°、11.8±0.2°、13.1±0.2°、27.6±0.2°、10.9±0.2°、13.3±0.2°、21.2±0.2°、15.8±0.2°、17.5±0.2°； The X-ray powder diffraction pattern of the isethionate salt form II comprises one or more diffraction peaks located at 2θ of 21.7±0.2°, 10.0±0.2°, 8.8±0.2°, 19.3±0.2°, 27.6±0.2°, 10.9±0.2°, 23.8±0.2°, 16.7±0.2°, 15.4±0.2°, 15.8±0.2°, 17.5±0.2°, 14.7±0.2°, 24.4±0.2°, 27.3±0.2°, and 29.2±0.2°, preferably, 4, 5, 6, 8, or 10 of them have diffraction peaks; for example, 10.0±0.2°、8.8±0.2°、19.3±0.2°、29.2±0.2°； 21.7±0.2°、10.0±0.2°、8.8±0.2°、19.3±0.2°、27.6±0.2°、29.2±0.2°； 21.7±0.2°、8.8±0.2°、19.3±0.2°、27.6±0.2°、10.9±0.2°、23.8±0.2°、27.3±0.2°、17.5±0.2°； 10.0±0.2°、8.8±0.2°、19.3±0.2°、27.6±0.2°、10.9±0.2°、23.8±0.2°、16.7±0.2°、15.4±0.2°、15.8±0.2°、17.5±0.2°； The X-ray powder diffraction pattern of the isethionate salt form III comprises one or more diffraction peaks located at 2θ of 19.4±0.2°, 16.9±0.2°, 26.6±0.2°, 14.6±0.2°, 28.0±0.2°, 25.6±0.2°, 20.7±0.2°, 12.8±0.2°, 19.1±0.2°, 27.2±0.2°, 24.4±0.2°, 15.3±0.2°, 26.2±0.2°, 30.2±0.2°, and 27.4±0.2°, preferably, comprising 4, 5, 6, 8 or 10 diffraction peaks selected therefrom; for example, 19.4±0.2°、16.9±0.2°、26.6±0.2°、14.6±0.2°； 19.4±0.2°、16.9±0.2°、26.6±0.2°、14.6±0.2°、28.0±0.2°、27.4±0.2°； 19.4±0.2°、16.9±0.2°、26.6±0.2°、14.6±0.2°、28.0±0.2°、25.6±0.2°、20.7±0.2°、27.4±0.2°； 19.4±0.2°、16.9±0.2°、26.6±0.2°、14.6±0.2°、28.0±0.2°、25.6±0.2°、20.7±0.2°、12.8±0.2°、19.1±0.2°、27.2±0.2°； The X-ray powder diffraction pattern of the sulfate crystalline form I comprises one or more diffraction peaks located at 2θ of 19.0±0.2°, 19.4±0.2°, 12.4±0.2°, 26.2±0.2°, 17.6±0.2°, 18.1±0.2°, 25.3±0.2°, 8.8±0.2°, 21.9±0.2°, and 11.5±0.2°, preferably, comprising 4, 5, 6, 8 or 10 diffraction peaks selected therefrom; for example, 19.0±0.2°、19.4±0.2°、12.4±0.2°、26.2±0.2°； 19.0±0.2°、19.4±0.2°、12.4±0.2°、26.2±0.2°、17.6±0.2°、11.5±0.2°； 19.0±0.2°、19.4±0.2°、12.4±0.2°、26.2±0.2°、17.6±0.2°、18.1±0.2°、25.3±0.2°、8.8±0.2°； 19.0±0.2°、19.4±0.2°、12.4±0.2°、26.2±0.2°、17.6±0.2°、18.1±0.2°、25.3±0.2°、8.8±0.2°、21.9±0.2°、11.5±0.2°； The X-ray powder diffraction pattern of sulfate crystalline form II comprises one or more diffraction peaks located at 2θ of 15.5±0.2°, 11.1±0.2°, 8.9±0.2°, 19.3±0.2°, 22.3±0.2°, 23.6±0.2°, 17.4±0.2°, 27.3±0.2°, 17.0±0.2°, 27.9±0.2°, 15.8±0.2°, 24.2±0.2°, 21.8±0.2°, 10.3±0.2°, and 20.6±0.2°, preferably, comprising 4, 5, 6, 8 or 10 diffraction peaks selected therefrom; for example, 11.1±0.2°、8.9±0.2°、19.3±0.2°、21.8±0.2°； 11.1±0.2°、8.9±0.2°、19.3±0.2°、22.3±0.2°、20.6±0.2°、27.9±0.2°； 15.5±0.2°、11.1±0.2°、8.9±0.2°、22.3±0.2°、23.6±0.2°、17.4±0.2°、20.6±0.2°、27.9±0.2°； 11.1±0.2°、8.9±0.2°、19.3±0.2°、22.3±0.2°、23.6±0.2°、17.4±0.2°、27.3±0.2°、17.0±0.2°、27.9±0.2°、20.6±0.2°； The X-ray powder diffraction pattern of sulfate crystalline form III comprises one or more diffraction peaks located at 2θ of 19.6±0.2°, 18.0±0.2°, 18.4±0.2°, 16.8±0.2°, 14.3±0.2°, 11.8±0.2°, 14.9±0.2°, 25.7±0.2°, 15.4±0.2°, 23.5±0.2°, 18.8±0.2°, 24.7±0.2°, 9.5±0.2°, 8.8±0.2°, and 11.1±0.2°, preferably, comprising 4, 5, 6, 8 or 10 diffraction peaks selected therefrom; for example, 19.6±0.2°、18.0±0.2°、18.4±0.2°、16.8±0.2°； 19.6±0.2°、18.0±0.2°、18.4±0.2°、16.8±0.2°、14.3±0.2°、11.1±0.2°； 19.6±0.2°、18.0±0.2°、18.4±0.2°、16.8±0.2°、14.3±0.2°、11.8±0.2°、14.9±0.2°、11.1±0.2°； 19.6±0.2°、18.0±0.2°、18.4±0.2°、16.8±0.2°、14.3±0.2°、11.8±0.2°、14.9±0.2°、25.7±0.2°、15.4±0.2°、23.5±0.2°； The X-ray powder diffraction pattern of sulfate salt form IV comprises one or more diffraction peaks located at 2θ of 19.4±0.2°, 18.9±0.2°, 15.5±0.2°, 8.8±0.2°, 18.1±0.2°, 24.9±0.2°, 17.4±0.2°, 12.3±0.2°, 26.1±0.2°, 14.5±0.2°, 22.2±0.2°, 24.3±0.2°, 21.7±0.2°, and 23.6±0.2°, preferably, comprising 4, 5, 6, 8 or 10 diffraction peaks selected therefrom; for example, 19.4±0.2°、18.9±0.2°、15.5±0.2°、8.8±0.2°； 19.4±0.2°、18.9±0.2°、15.5±0.2°、8.8±0.2°、18.1±0.2°、23.6±0.2°； 19.4±0.2°、18.9±0.2°、15.5±0.2°、8.8±0.2°、18.1±0.2°、24.9±0.2°、17.4±0.2°、23.6±0.2°； 19.4±0.2°、18.9±0.2°、15.5±0.2°、8.8±0.2°、18.1±0.2°、24.9±0.2°、17.4±0.2°、12.3±0.2°、26.1±0.2°、14.5±0.2°。 12. An acid salt of the compound according to any one of claims 8 to 11, Features: The X-ray powder diffraction pattern of the isethionate salt form I is shown in FIG1 ; The X-ray powder diffraction pattern of the isethionate salt form II is shown in FIG4 ; The X-ray powder diffraction pattern of the isethionate salt form III is shown in FIG7 ; The X-ray powder diffraction pattern of sulfate crystal form I is shown in Figure 10; The X-ray powder diffraction pattern of sulfate crystal form II is shown in Figure 11; The X-ray powder diffraction pattern of sulfate crystal form III is shown in Figure 12; The X-ray powder diffraction pattern of sulfate crystal form IV is shown in FIG13 .

13. An acid salt of the compound according to any one of claims 8 to 11, Features: The 2θ error between the top ten diffraction peak positions with the highest relative peak intensity in the X-ray powder diffraction pattern of the isethionate salt form I and the diffraction peak at the corresponding position in FIG. 1 is ±0.2° to ±0.5°, preferably ±0.2° to ±0.3°, and most preferably ±0.2°; The 2θ error between the top ten diffraction peak positions with the highest relative peak intensity in the X-ray powder diffraction pattern of the isethionate salt form II and the diffraction peak at the corresponding position in FIG. 4 is ±0.2° to ±0.5°, preferably ±0.2° to ±0.3°, and most preferably ±0.2°; The 2θ error between the top ten diffraction peak positions with the highest relative peak intensity in the X-ray powder diffraction pattern of the isethionate salt form III and the diffraction peak at the corresponding position in FIG. 7 is ±0.2° to ±0.5°, preferably ±0.2° to ±0.3°, and most preferably ±0.2°; The 2θ error between the top ten diffraction peak positions with the highest relative peak intensity in the X-ray powder diffraction pattern of sulfate crystal form I and the diffraction peak at the corresponding position in FIG. 10 is ±0.2° to ±0.5°, preferably ±0.2° to ±0.3°, and most preferably ±0.2°; The 2θ error between the top ten diffraction peak positions with the highest relative peak intensity in the X-ray powder diffraction pattern of sulfate crystal form II and the diffraction peak at the corresponding position in FIG. 11 is ±0.2° to ±0.5°, preferably ±0.2° to ±0.3°, and most preferably ±0.2°; The 2θ error between the top ten diffraction peak positions with the highest relative peak intensity in the X-ray powder diffraction pattern of sulfate crystal form III and the diffraction peak at the corresponding position in FIG. 12 is ±0.2° to ±0.5°, preferably ±0.2° to ±0.3°, and most preferably ±0.2°; The 2θ error between the top ten diffraction peak positions with relative peak intensity in the X-ray powder diffraction pattern of sulfate crystal form IV and the diffraction peak at the corresponding position in Figure 13 is ±0.2° to ±0.5°, preferably ±0.2° to ±0.3°, and most preferably ±0.2°.

14. An acid salt of the compound according to any one of claims 1 to 13, It is characterized in that The isethionate salt form I has a DSC spectrum as shown in FIG2 ; Isethionate salt Form II has a DSC spectrum as shown in Figure 5; The isethionate salt form III has a DSC spectrum as shown in FIG8 .

15. An acid salt of the compound according to any one of claims 1 to 14, It is characterized in that The acid salt crystal form is a hydrate or anhydrous. When the acid salt crystal form is a hydrate, the number of water is 0.2-3, preferably 0.2, 0.5, 1, 1.5, 2, 2.5 or 3, and more preferably 0.5, 1, 2 or 3; further, the water in the hydrate is pipeline water or crystal water or a combination of the two.

16. A method for preparing an acid salt of the compound according to any one of claims 1 to 15, comprising the steps of: 1) Weigh an appropriate amount of free base and add a reaction solvent to dissolve it; 2) adding an appropriate amount of acid and stirring; the amount of acid is preferably 1.2 equivalents; 3) After centrifugal drying, an acid salt of the compound or a crystalline form thereof is obtained; The reaction solvent is an organic solvent, preferably at least one of ethanol, 2-methyltetrahydrofuran, n-heptane, methyl tert-butyl ether, toluene, isopropyl acetate, tert-butanol, n-butanol, tetrahydrofuran, acetone, 2-butanone, ethyl acetate or 1,4-dioxane; The acid is selected from hydrochloric acid, sulfuric acid, nitric acid, hydrobromic acid, hydrofluoric acid, hydroiodic acid, phosphoric acid, 2,5-dihydroxybenzoic acid, 1-hydroxy-2-naphthoic acid, acetic acid, ethanesulfonic acid, dichloroacetic acid, trichloroacetic acid, acetohydroxamic acid, adipic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, benzoic acid, 4-acetamidobenzoic acid, 4-aminobenzoic acid, capric acid, caproic acid, caprylic acid, cinnamic acid, citric acid, cyclohexanesulfamic acid, camphorsulfonic acid, aspartic acid, camphoric acid, gluconic acid, glucuronic acid, glutamic acid, isoascorbic acid, lactic acid, malic acid, mandelic acid, pyroglutamic acid, tartaric acid, dodecylsulfuric acid, dibenzoyltartaric acid, ethane-1,2-disulfonic acid , ethanesulfonic acid, formic acid, fumaric acid, galactosonic acid, gentisic acid, glutaric acid, 2-ketoglutaric acid, glycolic acid, hippuric acid, isethionic acid, lactobionic acid, ascorbic acid, aspartic acid, lauric acid, camphoric acid, maleic acid, malonic acid, methanesulfonic acid, 1,5-naphthalenedisulfonic acid, naphthalene-2-sulfonic acid, nicotinic acid, oleic acid, orotic acid, oxalic acid, palmitic acid, pamoic acid, propionic acid, salicylic acid, 4-aminosalicylic acid, sebacic acid, stearic acid, succinic acid, thiocyanic acid, undecylenic acid, trifluoroacetic acid, benzenesulfonic acid, p-toluenesulfonic acid or L-malic acid; preferably hydrochloric acid, phosphoric acid, ethanesulfonic acid, benzenesulfonic acid, methanesulfonic acid, fumaric acid, isethionic acid, oxalic acid or hydrobromic acid.

17. A method for preparing an acid salt of the compound according to any one of claims 1 to 15, comprising the steps of: 1) Weigh an appropriate amount of free base and add a reaction solvent to dissolve it; 2) Add appropriate amount of acid and organic solvent and stir to dissolve; 3) optionally, adding seed crystals; 4) cooling, filtering out the precipitated solid, washing with a solvent, and drying; The reaction solvent used in step 1) is an organic solvent, preferably at least one of ethanol, propanol, isopropanol, 2-methyltetrahydrofuran, n-heptane, methyl tert-butyl ether, toluene, isopropyl acetate, tert-butanol, n-butanol, tetrahydrofuran, acetone, 2-butanone, ethyl acetate or 1,4-dioxane; The acid in step 2) is selected from hydrochloric acid, sulfuric acid, nitric acid, hydrobromic acid, hydrofluoric acid, hydroiodic acid, phosphoric acid, 2,5-dihydroxybenzoic acid, 1-hydroxy-2-naphthoic acid, acetic acid, ethanesulfonic acid, dichloroacetic acid, trichloroacetic acid, acetohydroxamic acid, adipic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, benzoic acid, 4-acetamidobenzoic acid, 4-aminobenzoic acid, capric acid, caproic acid, caprylic acid, cinnamic acid, citric acid, cyclohexanesulfamic acid, camphorsulfonic acid, aspartic acid, camphoric acid, gluconic acid, glucuronic acid, glutamic acid, isoascorbic acid, lactic acid, malic acid, mandelic acid, pyroglutamic acid, tartaric acid, dodecyl sulfuric acid, dibenzoyltartaric acid, ethane-1,2- disulfonic acid, ethanesulfonic acid, formic acid, fumaric acid, galactosonic acid, gentisic acid, glutaric acid, 2-ketoglutaric acid, glycolic acid, hippuric acid, isethionic acid, lactobionic acid, ascorbic acid, aspartic acid, lauric acid, camphoric acid, maleic acid, malonic acid, methanesulfonic acid, 1,5-naphthalenedisulfonic acid, naphthalene-2-sulfonic acid, nicotinic acid, oleic acid, orotic acid, oxalic acid, palmitic acid, pamoic acid, propionic acid, salicylic acid, 4-aminosalicylic acid, sebacic acid, stearic acid, succinic acid, thiocyanic acid, undecylenic acid, trifluoroacetic acid, benzenesulfonic acid, p-toluenesulfonic acid or L-malic acid; preferably hydrochloric acid, phosphoric acid, ethanesulfonic acid, benzenesulfonic acid, methanesulfonic acid, fumaric acid, isethionic acid, oxalic acid or hydrobromic acid; The organic solvent in step 2) is selected from one or more of alcohols, ethers, ketones or esters, preferably at least one of ethanol, propanol, isopropanol, 2-methyltetrahydrofuran, n-heptane, methyl tert-butyl ether, toluene, isopropyl acetate, tert-butanol, n-butanol, tetrahydrofuran, acetone, 2-butanone, ethyl acetate or 1,4-dioxane; The solvent in step 3) is selected from one or more of alcohols, ethers, ketones or ester solvents, preferably at least one of ethanol, propanol, isopropanol, 2-methyltetrahydrofuran, n-heptane, methyl tert-butyl ether, toluene, isopropyl acetate, tert-butanol, n-butanol, tetrahydrofuran, acetone, 2-butanone, ethyl acetate or 1,4-dioxane.

18. A pharmaceutical composition comprising a therapeutically effective amount of an acid salt of a compound according to any one of claims 1 to 15, and one or more pharmaceutically acceptable carriers, diluents or excipients.

19. Use of the acid salt of the compound according to any one of claims 1 to 15 or the pharmaceutical composition according to claim 18 in the preparation of a KRAS inhibitor drug; preferably in the preparation of a RAS G12C mutation inhibitor drug.

20. Use of an acid salt of a compound according to any one of claims 1 to 17 or a pharmaceutical composition according to claim 18 in the preparation of a medicament for treating Noonan syndrome, Leopard syndrome, leukemia, neuroblastoma, melanoma, esophageal cancer, head and neck tumors, breast cancer, lung cancer and colon cancer; preferably non-small cell lung cancer, colon cancer, esophageal cancer and head and neck tumors.