Pyridone compounds and preparation methods and applications thereof

Abstract

The present invention discloses a pyridone compound, a preparation method and an application thereof. Specifically, the present invention relates to a compound of formula I, which can be used to treat diseases related to irritable bowel syndrome (IBS) and other gastrointestinal disorders, as well as to treat RET and / or TRK functional disorders or cancers that regulate RET and / or TRK activity, inflammatory bowel disease, neurodegenerative diseases, chronic pain, acute pain, inflammatory diseases, Trypanosoma cruzi infection, and diseases related to imbalanced bone remodeling regulation. #imgabs0#

Description

Technical Field

[0001] The present invention belongs to the field of medicine, and in particular, relates to pyridone compounds and preparation methods and applications thereof. Background Art

[0002] Rearranged during transfection (RET) is a nerve growth factor receptor tyrosine kinase that plays an important role in the development of the kidney and enteric nervous system, and in the homeostasis maintenance of the nervous, endocrine, hematopoietic, and male reproductive systems. The structure of RET is divided into an extracellular region, a transmembrane region, and an intracellular kinase region. Its signal transduction is mediated by the binding of soluble proteins of the glial cell line-derived neurotrophin (GDNF) and the family ligand (GFL). Its ligand neurotrophin (GDNF) family does not bind directly to RET, but first forms a complex GFL-GFRα with the GDNF family receptor α, and then catalyzes RET homodimerization, causing RET to autophosphorylate in the intracellular region, and then recruits adapter proteins and pathway proteins to activate multiple signaling pathways including MAPK, PI3K, JAK-STAT, PKA, and PKC, thereby participating in cell proliferation, nerve conduction, cell migration, and cell differentiation (Alexander Drilon, Nature Reviews Clinical Oncology, 2018, 15: 151–167). These transduced messages play an important role in regulating cell survival, differentiation, proliferation, migration and chemotaxis.

[0003] RET plays an important role in the growth and survival of afferent pain receptors in the skin and intestine. RET kinase knockout mice lack enteric neurons and have other nervous system abnormalities, suggesting that a functional RET kinase protein product may be required for normal development of the nervous system (Taraviras, S. et al., 1999, 126: 2785-2797). In a population study of patients with Hirschsprung disease characterized by colonic obstruction, a high proportion of familial and sporadic RET deletion mutations appeared (Butler Tjaden N., et al., Transl. Res., 2013, 162: 1-15).

[0004] Chromosomal rearrangement of the RET gene may cause the RET gene to break. After the break, the 3' end of the RET gene can fuse with different genes such as KIF5B, TRIM33, CCDC6 or NCOA4 to form a fusion gene. The expressed fusion protein is continuously activated and drives the occurrence of tumors. It is reported that RET gene fusion exists in about 10-20% of PTC (papillary thyroid carcinoma) patients, mainly CCDC6-RET and NCOA4-RET fusion. About 1% to 2% of lung adenocarcinoma patients have RET fusion genes in their bodies, mainly KIF5B-RET, CCDC6-RET, TRIM33-RET, and NCOA4-RET, among which KIF5B-RET is the most common (Rosell R, and Karachaliou N, Lancet Oncol., 2016, 17: 1623-1625).

[0005] The gene encoding RET protein is located on the long arm of human chromosome 10. Its abnormalities (gene fusion, mutation, etc.) can cause a variety of diseases, including papillary thyroid carcinoma (PTC), medullary thyroid carcinoma (MTC), congenital medullary colon, lung adenocarcinoma, irritable bowel syndrome, etc.

[0006] Currently, there are two RET targeted drugs on the market, namely Loxo Oncology's LOXO-292 (selpercatinib / LY3527723) and Blurprint's BLU-667 (pralsetinib / Gavreto). These two targeted drugs have shown ideal efficacy and safety for patients with RET fusion or mutation-positive, especially RET fusion-positive non-small cell lung cancer (NSCLC) and RET mutation-positive medullary thyroid cancer (MTC).

[0007] Trk is a high-affinity receptor tyrosine kinase activated by a group of soluble growth factors called neurotrophic factors (NT). The Trk receptor family has three members, namely Trk A, Trk B and Trk C. Trk A is a high-affinity receptor for nerve growth factor (hereinafter referred to as NGF), Trk B is a high-affinity receptor for brain-derived neurotrophic factor (BDNF) and neurotrophic factor (hereinafter referred to as NT)-4 / 5, and Trk C is a high-affinity receptor for NT-3. All Trk receptors are highly expressed in neural tissue and participate in the differentiation and maintenance of nerve cell function.

[0008] Trk has been reported to be involved in nociceptive pain of osteoarthritis, chronic low back pain, rheumatoid arthritis, fractures, interstitial cystitis and chronic pancreatitis, neuropathic pain, and cancer pain associated with both of the above pain types. In addition, Trk receptors are expressed in cancer cells (e.g., glioma, hepatobiliary carcinoma, papillary thyroid carcinoma, colon cancer, non-small cell lung cancer, head and neck squamous cell carcinoma, pancreatic cancer, sarcoma and melanoneuroblastoma, prostate cancer and pancreatic cancer), inflammatory cells (e.g., mast cells and eosinophils), immunocompetent cells (e.g., T cells and B cells) and keratinocytes, and are reported to be possibly involved in the proliferation, migration and metastasis of cancer cells, inflammatory diseases (e.g., ulcerative colitis and Crohn's disease), allergic diseases (e.g., asthma, rhinitis and atopic dermatitis) and other diseases (e.g., psoriasis).

[0009] Therefore, compounds having Trk inhibitory activity are useful in the treatment of nociceptive pain, neuropathic pain and pain combining these two types of pain, cancer, inflammatory diseases, allergic diseases and psoriasis.

[0010] Irritable bowel syndrome (IBS) is a common functional disease that affects 10-20% of individuals and is characterized by abnormal bowel movements, bloating, abdominal pain, and visceral hypersensitivity. Currently, most studies believe that the cause of IBS is due to disorders between the brain and the gastrointestinal tract, interference with intestinal microorganisms, or increased inflammation, which affects intestinal transport function, leading to diarrhea and constipation. Some patients take medications at will, which further leads to intestinal flora imbalance and aggravates the disease. These studies suggest that inhibiting RET and / or TRK may be an effective strategy for the treatment of diseases related to irritable bowel syndrome (IBS), inflammatory bowel disease, other gastrointestinal disorders, neurodegenerative diseases, chronic pain, acute pain, inflammatory diseases, and the treatment of RET and / or TRK functional disorders or cancers that regulate RET and / or TRK activity. Summary of the invention

[0011] The present invention aims to provide a RET and TRK kinase inhibitor, which can be used to treat diseases related to irritable bowel syndrome (IBS), inflammatory bowel disease, other gastrointestinal disorders, neurodegenerative diseases, chronic pain, acute pain, inflammatory diseases, and to treat RET and / or TRK functional disorders or cancers that regulate RET and / or TRK activity.

[0012] In a first aspect of the present invention, there is provided a compound of formula I or a pharmaceutically acceptable salt, hydrate, solvate, isotope compound or prodrug thereof;

[0013]

[0014] In the formula,

[0015] Each R1 is independently H, halogen or selected from substituted or unsubstituted hydroxy, amino, C1-C6 alkyl, C3-C6 cycloalkyl, C6-C10 aryl, 5-10 membered heteroaryl,

[0016] R', R"" are each independently selected from substituted or unsubstituted C1-C6 alkyl, C3-C6 cycloalkyl, 3-6 membered heterocyclyl, C6-C10 aryl, 5-10 membered heteroaryl, wherein the substitution refers to substitution by one or more groups selected from the group consisting of halogen, hydroxyl, C1-C6 alkyl;

[0017] or when two R1 are connected to two adjacent atoms on the ring, they may be fused to form a substituted or unsubstituted C4-C8 cycloalkenyl, 4-8 membered heterocyclyl, C6-C10 aryl, 5-10 membered heteroaryl;

[0018] Each R2 is independently H, halogen, cyano, nitro or selected from substituted or unsubstituted hydroxyl, amino, C1-C6 alkyl, C3-C6 cycloalkyl;

[0019] or when two R2 are attached to two adjacent atoms on the ring, they may be fused to form a substituted or unsubstituted C4-C8 cycloalkenyl, 4-8 membered heterocyclyl, C6-C10 aryl, 5-10 membered heteroaryl;

[0020] Each R b are independently halogen, cyano, nitro or selected from substituted or unsubstituted amino, C1-C6 alkyl, C3-C6 cycloalkyl, 3-8 membered cycloheteroalkyl, C6-C10 aryl, 5-10 membered heteroaryl, Wherein, the substitution refers to the substitution of one or more R a replace:

[0021] Each R a independently selected from halogen, cyano, hydroxyl, C1-C6 alkyl, halogenated C1-C6 alkyl, C1-C6 alkoxy, halogenated C1-C6 alkoxy,

[0022] R6, R7 and R'7 are each independently selected from H, C1-C6 alkyl, halogenated C1-C6 alkyl, C3-C6 cycloalkyl, halogenated C3-C6 cycloalkyl, 3-8 membered cycloheteroalkyl;

[0023] R5 is hydrogen, cyano, halogen, nitro or selected from substituted or unsubstituted aldehyde, amino, hydroxyl, C1-C6 alkyl, 3-8 membered heteroalkyl, C1-C6 alkoxy, C3-C6 cycloalkyl, 3-8 membered heterocyclyl, C2-C6 alkenyl, C2-C6 alkynyl, -(L1)p-OH, -(L1)p-(C1-C6 alkoxy),

[0024] or R b When R and R5 are located at two adjacent C atoms, the C atoms to which they are connected are fused to form a substituted or unsubstituted C4-C8 cycloalkenyl, 4-8 membered cycloheteroyl, C6-C10 aryl, or 5-10 membered heteroaryl;

[0025] R8, R 10 , R' 10 , R 11 , R' 11 and R” 11 Each is independently H or selected from substituted or unsubstituted C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl, 3-6 membered cycloheteroalkyl, C6-C10 aryl, 5-10 membered heteroaryl, wherein the substitution refers to substitution by one or more groups selected from the following groups: halogen, hydroxyl, C1-C6 alkyl, N(R' 13 )(R” 13 );R' 13 and R” 13 Each is independently selected from H, halogen, C1-C6 alkyl;

[0026] or R 10 and R' 10 Together with the N atom to which it is attached, it forms a substituted or unsubstituted 3-8 membered heterocyclic group;

[0027] or R' 11 and R” 11 Together with the N atom to which it is attached, it forms a substituted or unsubstituted 3-8 membered heterocyclic group;

[0028] R9 and R'9 are each independently H, cyano or selected from substituted or unsubstituted aldehyde, C1-C6 alkyl, Hydroxyl, 3-8 membered heteroalkyl, 3-8 membered heterocyclyl, C6-C10 aryl, 5-10 membered heteroaryl, wherein R 12 and R' 12 Each is independently selected from C1-C6 alkyl, halogenated C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl, C3-C6 cycloheteroalkyl;

[0029] or R9 and R'9 together with the N atom to which they are attached form a substituted or unsubstituted 3-8 membered heterocyclic group;

[0030] Unless otherwise specified, the substitution refers to substitution by one or more R;

[0031] Each R is independently selected from halogen, cyano, aldehyde, hydroxyl, amino, C1-C6 alkyl, C1-C6 alkoxy;

[0032] Each L1 and L2 is independently selected from: CO, SO2; among them, R f and R g Each is independently selected from H, C1-C6 alkyl, halogen, cyano, hydroxyl, amino;

[0033] r is 1, 2, or 3;

[0034] m and p are each independently 1, 2, 3, 4, 5 or 6;

[0035] n is 0, 1 or 2;

[0036] n' is 0, 1, 2 or 3;

[0037] n” is 0, 1, 2 or 3.

[0038] In another preferred embodiment, Selected from in, is a single bond or a double bond; q is 0, 1 or 2; X1 and X2 are each independently selected from NH, O, S or CH2; X3 and X4 are each independently selected from N or CH; R1 is as defined above.

[0039] In another preferred embodiment, the compound of formula I or its pharmaceutically acceptable salt, hydrate, solvate, isotope compound or prodrug has a structure shown in formula I':

[0040]

[0041] In the formula,

[0042] R1 is selected from H, halogen, hydroxy, amino, C1-C6 alkyl, halo-C1-C6 alkyl, C1-C6 alkoxy, halo-C1-C6 alkoxy, C3-C6 cycloalkyl, halo-C3-C6 cycloalkyl, C3-C6 cycloalkyloxy, halo-C3-C6 cycloalkyloxy, (C1-C6 alkyl)NH-, (C1-C6 alkyl)(C1-C6 alkyl)N-;

[0043] Each R2 is independently selected from H, halogen, hydroxyl, amino, cyano, nitro, C1-C6 alkyl, halogenated C1-C6 alkyl, C3-C6 cycloalkyl, halogenated C3-C6 cycloalkyl;

[0044] R3 and R4 are each independently selected from H, halogen, hydroxyl, amino, cyano, C1-C6 alkyl, halogenated C1-C6 alkyl, C3-C6 cycloalkyl, 3-8 membered cycloheteroalkyl, C6-C10 aryl, 5-10 membered heteroaryl, wherein the C1-C6 alkyl group is optionally substituted by one or more groups selected from the group consisting of cyano, hydroxyl, C1-C6 alkoxy, halogenated C1-C6 alkoxy, The C6-C10 aryl or 5-10 membered heteroaryl is optionally substituted by one or more groups selected from the group consisting of halogen, C1-C6 alkyl, halogenated C1-C6 alkyl;

[0045] R6, R7 and R'7 are each independently selected from H, C1-C6 alkyl, halogenated C1-C6 alkyl, C3-C6 cycloalkyl, halogenated C3-C6 cycloalkyl, 3-8 membered cycloheteroalkyl;

[0046] R5 is selected from: hydrogen, cyano, aldehyde, amino, hydroxyl, halogen, C1-C6 alkyl, 3-8 membered heteroalkyl, halogenated C1-C6 alkyl, C1-C6 alkylene-OH, -(L1)p-(C1-C6 alkoxy), C1-C6 alkoxy, C3-C6 cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl,

[0047] R8, R 10 , R' 10 , R 11 , R' 11 and R” 11 Each is independently selected from H, C1-C6 alkyl, halogenated C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl, 3-6 membered cycloheteroalkyl, C6-C10 aryl, 5-10 membered heteroaryl;

[0048] or R 10 and R' 10 The nitrogen atom to which it is connected forms a substituted or unsubstituted 3-8 membered heterocyclic group, wherein the substitution refers to substitution by one or more groups selected from the group consisting of halogen, cyano, -CHO, hydroxyl, amino, C1-C6 alkyl, C1-C6 alkoxy, N(R' 13 )(R” 13 );R' 13 and R” 13 Each is independently selected from H, halogen or C1-C6 alkyl;

[0049] R9 and R'9 are each independently selected from cyano, aldehyde, C1-C6 alkyl, Hydroxyl, 3-8 membered heteroalkyl, 3-8 membered heterocyclyl, 5-10 membered heteroaryl, wherein R12 and R' 12 Each is independently selected from C1-C6 alkyl, halogenated C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl, C3-C6 cycloheteroalkyl;

[0050] or R9 and R'9 together with the N atom to which they are connected form a substituted or unsubstituted 3-8 membered heterocyclic group, wherein the substitution refers to substitution by one or more groups selected from the group consisting of halogen, cyano, aldehyde, hydroxyl, amino, C1-C6 alkyl, C1-C6 alkoxy;

[0051] L1 and L2 are independently Among them, the R f and R g Each is independently selected from H, C1-C6 alkyl, halogen, cyano, hydroxyl, amino;

[0052] n is 0, 1 or 2;

[0053] m and p are each independently 1, 2, 3, 4, 5 or 6.

[0054] In another preferred embodiment, R1 is selected from: C1-C6 alkoxy, halogenated C1-C6 alkoxy, C3-C6 cycloalkoxy, halogenated C3-C6 cycloalkoxy;

[0055] R5 is selected from cyano, aldehyde, C1-C6 alkyl, halogenated C1-C6 alkyl, 3-8 membered heteroalkyl, -C1-C6 alkylene-OH, C2-C6 alkenyl, C2-C6 alkynyl, -(L1)p-(C1-C6 alkoxy),

[0056]

[0057] R8, R 10 , R' 10 , R 11 , R' 11 and R” 11 Each is independently selected from H, C1-C6 alkyl, halogenated C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl, C3-C6 cycloheteroalkyl, C6-C10 aryl, 5-10 membered heteroaryl;

[0058] or R 10 and R' 10 The nitrogen atom to which it is connected forms a substituted or unsubstituted 3-8 membered heterocyclic group, wherein the substitution refers to substitution by one or more groups selected from the group consisting of halogen, cyano, aldehyde, hydroxyl, amino, C1-C6 alkyl, C1-C6 alkoxy, N(R' 13 )(R” 13 );R'13 and R” 13 Each is independently selected from H, halogen, C1-C6 alkyl;

[0059] R9 and R'9 are each independently selected from cyano, aldehyde, C1-C6 alkyl, Hydroxyl, 3-8 membered heteroalkyl, 3-8 membered heterocyclyl, 5-10 membered heteroaryl, wherein R 12 and R' 12 Each is independently selected from: C1-C6 alkyl, halogenated C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl, C3-C6 cycloheteroalkyl;

[0060] or R9 and R'9 together with the N atom to which they are connected form a substituted or unsubstituted 3-8 membered heterocyclic group, wherein the substitution refers to substitution by one or more groups selected from the group consisting of halogen, cyano, aldehyde, hydroxyl, amino, C1-C6 alkyl, C1-C6 alkoxy;

[0061] L1 and L2 are independently Among them, the R f and R g Each is independently selected from H, C1-C6 alkyl, halogen, cyano, hydroxyl, amino;

[0062] m and p are each independently 1, 2, 3, 4, 5 or 6;

[0063] Each R2 is independently selected from halogen, cyano, nitro, C1-C6 alkyl, halogenated C1-C6 alkyl;

[0064] R3 and R4 are each independently selected from H, halogen, hydroxy, amino, cyano, C1-C6 alkyl, halogenated C1-C6 alkyl.

[0065] In another preferred embodiment, R1 is selected from: C1-C6 alkoxy, halogenated C1-C6 alkoxy, C3-C6 cycloalkoxy, halogenated C3-C6 cycloalkoxy.

[0066] In another preferred embodiment, R5 is selected from cyano, aldehyde, C1-C6 alkyl, halogenated C1-C6 alkyl, 3-8 membered heteroalkyl, -C1-C6 alkylene-OH, C2-C6 alkenyl, C2-C6 alkynyl, -(L1)p-(C1-C6 alkoxy),

[0067] R8, R 10 , R' 10 , R 11 , R' 11 and R” 11Each is independently selected from H, C1-C6 alkyl, halogenated C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl, C3-C6 cycloheteroalkyl, C6-C10 aryl, 5-10 membered heteroaryl;

[0068] or R 10 and R' 10 The nitrogen atom to which it is connected forms a substituted or unsubstituted 3-8 membered heterocyclic group, wherein the substitution refers to substitution by one or more groups selected from the group consisting of halogen, cyano, aldehyde, hydroxyl, amino, C1-C6 alkyl, C1-C6 alkoxy, N(R' 13 )(R” 13 );R' 13 and R” 13 Each is independently selected from H, halogen, C1-C6 alkyl;

[0069] R9 and R'9 are each independently selected from cyano, aldehyde, C1-C6 alkyl, Hydroxyl, 3-8 membered heteroalkyl, 3-8 membered heterocyclyl, 5-10 membered heteroaryl, wherein R 12 and R' 12 Each is independently selected from: C1-C6 alkyl, halogenated C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl, C3-C6 cycloheteroalkyl;

[0070] or R9 and R'9 together with the N atom to which they are connected form a substituted or unsubstituted 3-8 membered heterocyclic group, wherein the substitution refers to substitution by one or more groups selected from the group consisting of halogen, cyano, aldehyde, hydroxyl, amino, C1-C6 alkyl, C1-C6 alkoxy;

[0071] L1 and L2 are independently Among them, the R f and R g Each is independently selected from H, C1-C6 alkyl, halogen, cyano, hydroxyl, amino;

[0072] m and p are each independently 1, 2, 3, 4, 5 or 6.

[0073] In another preferred embodiment, each R2 is independently selected from halogen, cyano, nitro, C1-C6 alkyl, and halogenated C1-C6 alkyl.

[0074] In another preferred embodiment, R3 and R4 are each independently selected from H, halogen, hydroxyl, amino, cyano, C1-C6 alkyl, halogenated C1-C6 alkyl.

[0075] In another preferred embodiment, the compound of formula I has a structure shown in formula I":

[0076]

[0077] In the formula,

[0078] Rc is a substituted or unsubstituted C1-C6 alkyl group, wherein the substitution refers to substitution by one or more groups selected from halogen, cyano, and hydroxyl groups;

[0079] X is selected from halogen, cyano, nitro, halogenated C1-C6 alkyl;

[0080] R5 is as defined above.

[0081] In another preferred embodiment, R5 is selected from

[0082] In another preferred embodiment, R5 is Wherein, R8 is selected from methyl, ethyl, isopropyl and tert-butyl.

[0083] In another preferred embodiment, R5 is wherein p is 1, 2, or 3; and R9 and R'9 are as defined above.

[0084] In another preferred embodiment, R5 is Among them, R 10 and R' 10 Each is independently selected from H, methyl, ethyl, isopropyl, tert-butyl, C3-C6 cycloalkyl (such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl), 3-6 membered heterocycloalkane (such as tetrahydrofuranyl, tetrahydropyrrolyl, morpholinyl), or R 10 and R' 10 Together with the nitrogen atom to which it is attached, it forms a 5-6 membered heterocyclic group.

[0085] In another preferred embodiment, R5 is Among them, R' 11 and R” 11 Each is independently selected from H, methyl, ethyl, isopropyl, tert-butyl, C3-C6 cycloalkyl (such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl), 3-6 membered heterocycloalkane (such as tetrahydrofuranyl, tetrahydropyrrolyl, morpholinyl).

[0086] In another preferred embodiment, R1, R2, R3, R4, and R5 are specific groups corresponding to the specific compounds in the examples.

[0087] In another preferred embodiment, the compound is selected from

[0088]

[0089]

[0090] In another preferred embodiment, the compound is selected from the compounds shown in the Examples.

[0091] The second aspect of the present invention provides a pharmaceutical composition comprising the compound described in the first aspect or a pharmaceutically acceptable salt, hydrate, solvate, isotope compound or prodrug thereof; and a pharmaceutically acceptable carrier or diluent.

[0092] In another preferred embodiment, the pharmaceutical composition further comprises one or more other therapeutic agents.The other therapeutic agent is selected from aminosalicylic acid preparations (such as sulfasalazine), glucocorticoids (such as hydrocortisone, dexamethasone, etc.), PD-1 inhibitors (such as nivolumab, pembrolizumab, JS-001, SHR-120, BGB-A317, IBI-308, GLS-010, GB-226, STW204, HX008, HLX10, BAT1306, AK105, LZM 009 or biosimilars of the above drugs, etc.), PD-L1 inhibitors (such as durvalumab, atezolizumab, CS1001, KN035, HLX20, SHR-1316, BGB-A333, JS003, CS1003, KL-A167, F 520, GR1405, MSB2311 or biosimilars of the above drugs, etc.), CD20 antibodies (such as rituximab, obinutuzumab, ofatumumab, tositumomab, ibritumomab tiuxetan, etc.), CD47 antibodies (such as Hu5F9-G4, CC-90002, TTI-621, TTI-622, OSE-172, SRF-231, ALX-148, NI-1701, SHR-1603, IBI188, IMM01), ALK inhibitors (such as ceritinib, alectinib, nivolumab, brigatinib, lorlatinib, oclatinib), PI3K inhibitors (such as idelalisib, dactolisib, taselisib, buparlisib, etc.), BTK inhibitors (such as ibrutinib, tirabrutinib, acalabrutinib, etc.), EGFR inhibitors (such as afatinib, gefitinib, erlotinib, lapatinib, dacomitinib, icotinib, canertinib, etc.), VEGFR inhibitors (such as sorafenib, pazopanib, rivatinib, cabozantinib, sunitinib, etc.), tinib, donafenib, etc.), HDAC inhibitors (such as givinostat, droxinostat, entinostat, dacilast, tekdin, etc.), CDK inhibitors (such as palbociclib, ribociclib, abemaciclib, lerociclib, etc.), MEK inhibitors (such as selumetinib (AZD6244), trametinib (GSK1120212), PD0325901, U0126, AS-703026, PD184352 (CI-1040), etc.) , Akt inhibitors (such as MK-2206, Ipatasertib, Capivasertib, Afuresertib, Uprosertib, etc.), mTOR inhibitors (such as Vistusertib, etc.), SHP2 inhibitors (such as RMC-4630, JAB-3068, TNO155, etc.), IGF-1R inhibitors (such as Ceritinib, Ocasartinib, linsitinib, BMS-754807, GSK1838705A, etc.). ,

[0093] The third aspect of the present invention provides a use of the compound described in the first aspect or its pharmaceutically acceptable salt, hydrate, solvate, isotope compound or prodrug in the preparation of a medicament for treating irritable bowel syndrome, other intestinal related diseases and / or cancer.

[0094] In another preferred embodiment, the pharmaceutically acceptable salt includes but is not limited to the following salts: acetate, adipate, alginate, ascorbate, aspartate, benzoate, benzenesulfonate, bisulfate, borate, butyrate, citrate, camphor salt, camphorsulfonate, cyclopentanepropionate, diglycolate, dodecyl sulfate, ethanesulfonate, fumarate, glucoheptonate, glycerophosphate, hemisulfate, heptanoate, hexanoate, hydrochloride, hydrobromide, hydroiodide, hydroxyethanesulfonate, lactate, maleate, methanesulfonate, naphthylsulfonate, nicotinate, nitrate, oxalate, pectinate, persulfate, phenylpropionate, phosphate, picrate, pivalate, propionate, salicylate, succinate, sulfate, sulfonate, tartrate, thiocyanate, toluenesulfonate, dodecanoate.

[0095] In another preferred embodiment, the irritable bowel syndrome is diarrhea-predominant irritable bowel syndrome, constipation-predominant irritable bowel syndrome, mixed irritable bowel syndrome, or unspecified irritable bowel syndrome.

[0096] In another preferred embodiment, the other intestinal related diseases are functional bloating, functional constipation, nonspecific functional intestinal disorder, functional abdominal pain syndrome, chronic idiopathic constipation, and functional gastroduodenopathy.

[0097] In another preferred embodiment, the irritable bowel syndrome includes: diarrhea-dominant type, constipation-dominant type or alternating defecation pattern, functional bloating, functional constipation, nonspecific functional intestinal disorder, functional abdominal pain syndrome, chronic idiopathic constipation, and functional gastroduodenopathy.

[0098] In another preferred embodiment, the cancer includes: bladder cancer, ovarian cancer, adenocarcinoma, gastric cancer, pancreatic cancer, prostate cancer, colon cancer, lung cancer, bone cancer, brain cancer, neuroblastoma, rectal cancer, familial adenomatous polyposis cancer, hereditary non-polyposis colorectal cancer, esophageal cancer, lip cancer, laryngeal cancer, hypopharyngeal cancer, tongue cancer, salivary gland cancer, medullary thyroid cancer, papillary thyroid cancer, kidney cancer, renal parenchymal cancer, cervical cancer, uterine body cancer, endometrial cancer, choriocarcinoma, testicular cancer, urinary cancer, melanoma, acute lymphocytic leukemia, chronic lymphocytic leukemia, acute myeloid leukemia, chronic myeloid leukemia, hepatocellular carcinoma, gallbladder cancer, bronchial cancer, small cell lung cancer, non-small cell lung cancer, and multiple myeloma.

[0099] In another preferred embodiment, the cancer is non-small cell lung cancer, glioma, multiple myeloma, hepatobiliary carcinoma, medullary thyroid carcinoma, papillary thyroid tumor, neuroblastoma, colon cancer, head and neck squamous cell carcinoma, pancreatic cancer, sarcoma, melanoma, fibrosarcoma, pancreatic tumor, soft tissue sarcoma, high-solid tumor, breast tumor or bile duct cancer.

[0100] In a fourth aspect, the present invention provides use of the compound described in the first aspect or a pharmaceutically acceptable salt, hydrate, solvate, isotope compound or prodrug thereof in the preparation of a medicament for treating neurodegenerative diseases, chronic pain, acute pain, inflammatory diseases, inflammatory bowel disease, Trypanosoma cruzi infection or diseases related to imbalance in bone remodeling regulation.

[0101] In a fifth aspect, the present invention provides a use of GSK-3179106 and / or BOS-589, or a pharmaceutically acceptable salt, hydrate, solvate, isotope compound or prodrug thereof in the preparation of a medicament for treating neurodegenerative diseases, chronic pain, acute pain, inflammatory diseases, inflammatory bowel disease, Trypanosoma cruzi infection or diseases associated with imbalanced regulation of bone remodeling;

[0102]

[0103] In the sixth aspect of the present invention, there is provided a use of the compound described in the first aspect or its pharmaceutically acceptable salt, hydrate, solvate, isotope compound or prodrug, or the pharmaceutical composition described in the second aspect in the preparation of a medicament for inhibiting RET kinase and / or TRK kinase activity in cells or subjects.

[0104] In a seventh aspect, the present invention provides a use of GSK-3179106 and / or BOS-589, or a pharmaceutically acceptable salt, hydrate, solvate, isotope compound or prodrug thereof in the preparation of a medicament for inhibiting RET kinase and / or TRK kinase activity in a cell or a subject.

[0105] In an eighth aspect, the present invention provides a method for treating RET and / or TRK related diseases, the method comprising administering to a subject identified or diagnosed as having RET and / or TRK related diseases a therapeutically effective amount of the compound as described in the first aspect or a pharmaceutically acceptable salt, hydrate, solvate, isotope compound or prodrug thereof, the pharmaceutical composition as described in the second aspect, GSK-3179106 or a pharmaceutically acceptable salt, hydrate, solvate, isotope compound or prodrug thereof, or BOS-589 or a pharmaceutically acceptable salt, hydrate, solvate, isotope compound or prodrug thereof.

[0106] In the ninth aspect of the present invention, a method for inhibiting RET and / or TRK kinase activity in a cell or a subject is provided, the method comprising the step of contacting the cell with or administering to the subject the compound or a pharmaceutically acceptable salt, hydrate, solvate, isotope compound or prodrug thereof described in the first aspect, the pharmaceutical composition described in the second aspect, GSK-3179106 or a pharmaceutically acceptable salt, hydrate, solvate, isotope compound or prodrug thereof, or BOS-589 or a pharmaceutically acceptable salt, hydrate, solvate, isotope compound or prodrug thereof.

[0107] In another preferred embodiment, the cell is a mammalian cell.

[0108] In another preferred embodiment, the subject is a mammal, preferably a human.

[0109] In a tenth aspect, the present invention provides a method for preparing the compound according to the first aspect or a pharmaceutically acceptable salt, hydrate, solvate, isotope compound or prodrug thereof, comprising the steps of:

[0110]

[0111] 1) reacting a compound of formula M with a compound of formula N to obtain a compound of formula A;

[0112]

[0113] 2) reacting the compound of formula A with an acid to obtain a compound of formula I;

[0114] wherein Y is selected from OH, halogen;

[0115] The other groups are as defined in the first aspect.

[0116] In an eleventh aspect of the present invention, there is provided a method for preparing a compound of formula 1-2, comprising the steps of:

[0117]

[0118] Compound 1-1 is slowly dropwise added with concentrated nitric acid or fuming nitric acid in the presence of concentrated sulfuric acid to carry out a nitration reaction to obtain compound 1-2, wherein Rb, n" are as defined in the first aspect.

[0119] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features specifically described below (such as embodiments) can be combined with each other to form a new or preferred technical solution. Due to space limitations, they will not be described one by one here. DETAILED DESCRIPTION

[0120] After extensive and in-depth research, the inventors unexpectedly discovered a class of SF5-substituted pyridone compounds that have good RET and TRK kinase inhibitory activity and good selectivity for VEGFR2 kinase. In addition, the compounds have good pharmacodynamics / pharmacokinetic properties. On this basis, the present invention was completed.

[0121] the term

[0122] In the present invention, unless otherwise specified, the terms used have the general meanings well known to those skilled in the art.

[0123] When substituents are described by conventional chemical formulas written from left to right, the substituents also include chemically equivalent substituents that would result if the formula were written from right to left. For example, -CH2O- is equivalent to -OCH2-.

[0124] The term "alkyl" by itself or as part of another substituent refers to a straight or branched hydrocarbon radical having the specified number of carbon atoms (i.e., C1-C6 refers to one to six carbon atoms). Examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, sec-butyl, n-pentyl, n-hexyl, and the like. One or more positions in the alkyl group are substituted, particularly 1 to 4 substituents, which may be substituted at any position.

[0125] The term "C1-C6 alkoxy" by itself or as part of another substituent refers to a straight chain, branched chain or cyclic alkoxy group (such as C3-C6 cycloalkoxy) having 1 to 6 carbon atoms, representative examples of which include (but are not limited to): methoxy, ethoxy, propoxy, isopropoxy and butoxy, etc. Preferably, it is C1-C3 alkoxy.

[0126] The term "heteroalkyl" by itself or as part of another substituent refers to a group in which the carbon atoms in the alkyl group are substituted by 1, 2 or 3 heteroatoms selected from N, O, S, Si or P, and wherein the nitrogen and sulfur atoms are optionally oxidized. In the present invention, "3-8 heteroalkyl" refers to a group containing 1-6 (i.e., 1, 2, 3, 4, 5 or 6) carbon atoms, and 1 or 2 heteroatoms selected from N, O, S or P. Representative examples include (but are not limited to): CH3OCH2-, CH3SCH2-, CH3CH2OCH2-, etc.

[0127] The term "cycloalkyl" itself or as part of another substituent refers to a cyclic alkyl group including a saturated monocyclic, bicyclic or polycyclic ring, such as a C3-C8 or C3-C12 cycloalkyl group. A C3-C8 cycloalkyl group refers to a C3, C4, C5, C6, C7 or C8 cycloalkyl group. Cycloalkyl groups may also include cycloalkyl groups having structures such as spirocyclic, bridged, and cyclic rings. Representative cycloalkyl groups of the present invention include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and norbornyl groups. It should be understood that substituted or unsubstituted cycloalkyl groups, such as branched cycloalkyl groups (such as 1-methylcyclopropyl and 2-methylcyclopropyl), are included in the definition of "cycloalkyl".

[0128] The term "cycloheteroalkyl" by itself or as part of another substituent refers to a cycloalkyl ring having the specified number of ring vertices (or members) and having one to five heteroatoms selected from N, O and S replacing carbon atoms in the ring backbone, wherein the nitrogen and sulfur atoms are optionally oxidized and the nitrogen atom is optionally quaternized. Cycloheteroalkyl is typically a 4-12 membered ring. Cycloheteroalkyl can be a monocyclic, bicyclic or polycyclic system. Examples of cycloheteroalkyl groups include, but are not limited to, pyrrolidinyl, imidazolidinyl, pyrazolidinyl, butyrolactamyl, valerolactamyl, imidazolidinone, hydantoinyl, dioxolanyl, phthalimidyl, piperidinyl, 1,4-dioxane, morpholinyl, thiomorpholinyl, thiomorpholine-S-oxide, thiomorpholine-S,S-oxide, piperazinyl, pyranyl, pyridone, 3-pyrrolinyl, thiopyranyl, pyranonyl, tetrahydrofuranyl, tetrahydrothiophenyl, quinuclidine, and the like.

[0129] The term "alkenyl" by itself or as part of another substituent refers to a straight or branched hydrocarbon group containing one or more double bonds and generally having a length of 2 to 20 carbon atoms (or C2-C8). For example, in the present invention, "C2-C6 alkenyl" contains two to six carbon atoms. Alkenyl includes, but is not limited to, for example, vinyl, propenyl, butenyl, 1-methyl-2-butene-1-yl, etc.

[0130] The term "aryl", alone or as part of a larger moiety such as "aralkyl", "aralkyloxy" or "aryloxyalkyl", refers to a monocyclic, bicyclic or tricyclic ring system (preferably 6-10 membered aromatic rings) having a total of 5 to 15 ring members, wherein at least one ring in the system is aromatic and wherein each ring in the system contains 3 to 7 ring members. "Aryl" can be substituted or unsubstituted. In certain embodiments of the present invention, "aryl" refers to an aromatic ring system, which includes but is not limited to phenyl, biphenyl, indanyl, 1-naphthyl, 2-naphthyl and tetrahydronaphthyl. Fused aryl groups may be attached to another group at a suitable position on the cycloalkyl ring or the aromatic ring. Connecting lines drawn from the ring system indicate that bonds may be attached to any suitable ring atom.

[0131] The term "heteroaryl" by itself or as part of another substituent refers to a heteroaromatic system containing 1 to 4 heteroatoms, 5 to 14 ring atoms, wherein the heteroatoms are selected from oxygen, nitrogen and sulfur. The heteroaryl group is preferably a 5-10-membered ring, more preferably a 5-membered or 6-membered ring, such as pyrrolyl, pyrazolyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, thiadiazolyl, isothiazolyl, furanyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, triazoxinyl, triazolyl and tetrazolyl. "Heteroaryl" may be substituted or unsubstituted, and when substituted, the substituent is preferably one or more of the following groups independently selected from alkyl, deuterated alkyl, haloalkyl, alkoxy, haloalkoxy, alkenyl, alkynyl, alkylthio, alkylamino, halogen, amino, nitro, hydroxy, mercapto, cyano, cycloalkyl, heterocyclyl, aryl, heteroaryl, cycloalkylthio, oxo, carboxyl and carboxylate.

[0132] Preferably, each "5-10 membered heteroaryl" is independently a 5-10 membered heteroaryl containing 1, 2 or 3 heteroatoms selected from N, O or S.

[0133] The term "heterocycle", "heterocyclyl" or "heterocyclic group" by itself or as part of another substituent refers to a stable 3-, 4-, 5-, or 7-membered monocyclic or bicyclic or 7-, 8-, 9-, 10-, 11-, 12-, 13- or 14-membered polycyclic heterocycle, including fused, spirocyclic and / or bridged ring structures, which are saturated, partially unsaturated or fully unsaturated, and which contain carbon atoms and 1, 2, 3 or 4 heteroatoms independently selected from N, O and S. The term also includes polycyclic groups formed by the fusion of heterocycles with aromatic rings (such as benzene rings). "Heterocycle" can be substituted or unsubstituted. The nitrogen and sulfur heteroatoms as ring atoms can be optionally oxidized. The nitrogen atom is substituted or unsubstituted (i.e., N or NR, wherein R is H or, if defined, another substituent). The heterocycle can be connected to its side group at any heteroatom or carbon atom that results in a stable structure. The heterocyclic groups described herein may be substituted on carbon or nitrogen atoms if the resulting compound is stable. The nitrogen in the heterocyclic ring may be optionally quaternized. Preferably, when the total number of S and O atoms in the heterocyclic ring exceeds 1, then these heteroatoms are not adjacent to each other. Preferably, the total number of S and O atoms in the heterocyclic ring is not greater than 1. When the term "heterocyclic ring" is used, it is intended to include heteroaryl groups.Examples of heterocycles include, but are not limited to, acridinyl, azetidinyl, azinyl, benzimidazolyl, benzofuranyl, benzothiofuranyl, benzothiophenyl, benzoxazolyl, benzoxazolinyl, benzothiazolyl, benzotriazolyl, benzotetrazolyl, benzisoxazolyl, benzisothiazolyl, benzimidazolinyl, carbazolyl, 4aH-carbazolyl, carbolinyl, chromanyl, chromenyl, cinnolinyl, decahydroquinolinyl, 2H,6H-1,5,2-dithiazinyl, dihydrofuro[2,3-b]tetrahydrofuranyl, furanyl, furazanyl, imidazolidinyl, imidazolinyl, imidazolyl, 1H- H-indazolyl, imidazopyridinyl, indolenyl, indolinyl, indolizinyl, indolyl, 3H-indolyl, isatinoyl, isobenzofuranyl, isochromanyl, isoindazolyl, isoindolinyl, isoindolyl, isoquinolinyl, isothiazolyl, isothiazolopyridinyl, isoxazolyl, isoxazolopyridinyl, methylenedioxyphenyl, morpholinyl, naphthyridinyl, octahydroisoquinolinyl, oxadiazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl oxazolyl, oxazolidinyl, oxazolyl, oxazolopyridinyl, oxazolidinyl, perylene, oxindolyl, pyrimidinyl, phenanthridinyl, phenanthrolinyl, phenazinyl, phenothiazinyl, phenoxathiyl, phenoxazinyl, phthalazinyl, piperazinyl, piperidinyl, piperidonyl, 4-piperidonyl, piperonyl, pteridinyl, purinyl, pyranyl, pyrazinyl, pyrazolidinyl, pyrazolinyl, pyrazolopyridinyl, pyrazolyl, pyridazinyl, pyridooxazolyl, pyridoimidazolyl, pyridothiazolyl, pyridinyl, pyrimidinyl, pyrrolidinyl, pyrrolinyl, 2-pyrrolidinonyl, 2H-pyrrolyl, pyrrolyl, quinoline The present invention also includes fused ring and spiro compounds containing heterocyclic rings such as the above.

[0134] Preferably, each "3-6 membered heterocyclyl" is a 3-6 membered heterocyclyl containing 1, 2 or 3 heteroatoms selected from N, O or S. The terms "4-8 membered heterocyclyl", "3-6 membered cycloheteroalkyl", "3-8 membered heterocyclyl", "4-8 membered cycloheteroalkyl", "C3-C6 cycloheteroalkyl" have similar meanings.

[0135] The term "C4-C8 cycloalkenyl" refers to a cyclic 4-8 membered ring containing 1, 2 or 3 olefinic bonds. In the present invention, each of the above-mentioned alkyl, haloalkyl, alkoxy, cycloalkyl, aryl, heteroaryl, cycloheteroalkyl, alkenyl, alkyne, heterocycle, heterocyclyl, etc., may be substituted or unsubstituted.

[0136] In the present invention, the term "substituted" refers to one or more hydrogen atoms on a specific group being replaced by a specific substituent. The specific substituent is the substituent described above, or the substituent appearing in each embodiment. Unless otherwise specified, a substituted group may have a substituent selected from a specific group at any substitutable site of the group, and the substituent may be the same or different at each position. It should be understood by those skilled in the art that the combination of substituents contemplated by the present invention is those that are stable or chemically feasible. Typical substitutions include, but are not limited to, one or more of the following groups: such as hydrogen, deuterium, halogen (e.g., monohalogen substituent or polyhalogen substituent, the latter such as trifluoromethyl or alkyl containing Cl3), nitrile, nitro, oxo (such as =O), trifluoromethyl, trifluoromethoxy, cycloalkyl, alkenyl, alkynyl, heterocycle, aromatic ring, -OR a 、-SR a 、-S(=O)R e 、-S(=O)2R e 、-P(=O)2R e 、-S(=O)2OR e ,

[0137] Among them, R a may independently represent hydrogen, deuterium, alkyl, cycloalkyl, alkenyl, alkynyl, heterocyclic or aromatic rings, and R b , R c and R d may independently represent hydrogen, deuterium, an alkyl group, a cycloalkyl group, a heterocyclic ring or an aromatic ring, or R b and R c Together with the N atom, it can form a heterocyclic ring; R e It can independently represent hydrogen, alkyl, cycloalkyl, alkenyl, alkynyl, heterocycle or aromatic ring. The above typical substituents, such as alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, heterocycle or aromatic ring can be optionally substituted. The substituents include (but are not limited to): halogen, hydroxyl, cyano, carboxyl (-COOH), C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C8 cycloalkyl, 3-12 membered heterocyclic group, aryl, heteroaryl, C1-C8 aldehyde group, C2-C10 acyl group, C2-C10 ester group, amine group, C1-C6 alkoxy group, C1-C10 sulfonyl group, and C1-C6 urea group.

[0138] Unless otherwise stated, any heteroatom with insufficient valence is assumed to have sufficient hydrogen atoms to complete its valence.

[0139] When a substituent is a non-terminal substituent, it is a substituent of the corresponding group, for example, alkyl for alkylene, cycloalkyl for cycloalkylene, heterocyclyl for heterocyclylene, alkoxy for alkyleneoxy, and the like.

[0140] The term "halo" or "halogen" includes fluorine, chlorine, bromine and iodine.

[0141] The term "PMB" refers to p-methoxybenzyl.

[0142] The term "aldehyde group" has the structure -CHO.

[0143] Active ingredients

[0144] As used herein, the term "compound of the present invention" or "active ingredient of the present invention" is used interchangeably to refer to a compound of formula I, or a pharmaceutically acceptable salt, hydrate, solvate, isotope compound (such as a deuterated compound) or prodrug thereof. The term also includes racemates and optical isomers.

[0145] In the present invention, the compound of formula I has the following structure:

[0146]

[0147] In the formula,

[0148] R1, R2, R5, R b , n, r, n' and n" are as defined above.

[0149] Preferably, the compound of formula I has a structure shown in formula I':

[0150]

[0151] In the formula, R1, R2, R3, R4, R5 and n are as defined above.

[0152] Preferably, in Formula I and Formula I', R1 is selected from C1-C6 alkoxy, halogenated C1-C6 alkoxy, C3-C6 cycloalkoxy, halogenated C3-C6 cycloalkoxy.

[0153] Preferably, in Formula I and Formula I', R5 is selected from cyano, aldehyde, C1-C6 alkyl, halogenated C1-C6 alkyl, 3-8 membered heteroalkyl, C1-C6 alkyl-OH, C2-C6 alkenyl, C2-C6 alkynyl,

[0154] R8, R 10 , R' 10 , R11 , R' 11 and R” 11 Each is independently selected from H, C1-C6 alkyl, halogenated C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl, C3-C6 cycloheteroalkyl, C6-C10 aryl, 5-10 membered heteroaryl;

[0155] or R 10 and R' 10 Together with the N atom to which it is connected, it forms a substituted or unsubstituted 3-8 membered heterocyclic group, wherein the substitution refers to substitution by one or more groups selected from the group consisting of halogen, cyano, -CHO, hydroxyl, amino, C1-C6 alkyl, C1-C6 alkoxy;

[0156] R9 and R'9 are each independently selected from: cyano, aldehyde, Hydroxyl, 3-8 membered heteroalkyl, 3-8 membered heterocyclyl, 5-10 membered heteroaryl, wherein R 12 and R' 12 Each is independently selected from: C1-C6 alkyl, halogenated C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl, C3-C6 cycloheteroalkyl;

[0157] or R9 and R'9 together with the N atom to which they are connected form a substituted or unsubstituted 3-8 membered heterocyclic group, wherein the substitution refers to substitution by one or more groups selected from the group consisting of halogen, cyano, aldehyde, hydroxyl, amino, C1-C6 alkyl, C1-C6 alkoxy;

[0158] L1 and L2 are independently Among them, the R f and R g Each is independently selected from H, C1-C6 alkyl, halogen, cyano, hydroxyl, amino;

[0159] m and p are each independently 1, 2, 3, 4, 5 or 6.

[0160] Preferably, in Formula I and Formula I', each R2 is independently selected from: halogen, cyano, nitro, C1-C6 alkyl, halogenated C1-C6 alkyl.

[0161] Preferably, the compound of formula I has a structure shown in formula I":

[0162]

[0163] In the formula,

[0164] Rc is selected from substituted or unsubstituted C1-C6 alkyl, wherein the substitution refers to substitution by one or more halogen, cyano, or hydroxyl groups;

[0165] X is selected from halogen, cyano, nitro, halogenated C1-C6 alkyl;

[0166] R5 is as defined above.

[0167] Preferably, in formula I, I', I", R5 is selected from

[0168] Preferably, in formula I, I', I", R5 is Wherein, R8 is selected from methyl, ethyl, isopropyl and tert-butyl.

[0169] Preferably, in formula I, I', I", R5 is wherein p is 1, 2, or 3; and R9 and R'9 are as defined above.

[0170] Preferably, in formula I, I', I", R5 is Among them, R 10 and R' 10 Each is independently selected from H, methyl, ethyl, isopropyl, tert-butyl, C3-C6 cycloalkyl (such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl), 3-6 membered heterocycloalkane (such as tetrahydrofuranyl, tetrahydropyrrolyl, morpholinyl), or R 10 and R' 10 Together with the nitrogen atom to which it is attached, it forms a 5-6 membered heterocyclic group.

[0171] Preferably, in formula I, I', I", R5 is Among them, R' 11 and R” 11 Each is independently selected from H, methyl, ethyl, isopropyl, tert-butyl, C3-C6 cycloalkyl (such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl), 3-6 membered heterocycloalkane (such as tetrahydrofuranyl, tetrahydropyrrolyl, morpholinyl).

[0172] Typically, the present invention provides a compound of formula I or a pharmaceutically acceptable salt, hydrate, solvate, isotope compound or prodrug thereof;

[0173]

[0174] In the formula,

[0175] Each R1 is independently selected from the following group: substituted or unsubstituted: H, halogen, hydroxyl, amino, C1-C6 alkyl, R'-O-, C3-C6 cycloalkyl, C6-C10 aryl, 5-10 membered heteroaryl, R'NH- or R'R"N-;

[0176] R', R" are each independently selected from the following substituted or unsubstituted groups: C1-C6 alkyl, C3-C6 cycloalkyl, 3-6 membered heterocyclyl, C6-C10 aryl, 5-10 membered heteroaryl, wherein the substitution refers to substitution by one or more groups selected from the following group: halogen, hydroxyl, C1-C6 alkyl;

[0177] Or when two R1 are connected to two adjacent atoms on the ring, they can be fused to form a substituted or unsubstituted group of the following groups: C4-C8 cycloalkenyl, 4-8 membered heterocyclyl, C6-C10 aryl, 5-10 membered heteroaryl;

[0178] Each R2 is independently selected from the following group: substituted or unsubstituted: H, halogen, hydroxyl, amino, cyano, nitro, R8C(O)O-, R8C(O)-, R8S(O)2, C1-C6 alkyl, C3-C6 cycloalkyl;

[0179] Or when two R2 are connected to two adjacent atoms on the ring, they can be fused to form a substituted or unsubstituted group of the following groups: C4-C8 cycloalkenyl, 4-8 membered heterocyclyl, C6-C10 aryl, 5-10 membered heteroaryl;

[0180] Each R b independently selected from the following groups substituted or unsubstituted: halogen, amino, cyano, nitro, R8C(O)O-, R8C(O)-, R8S(O)2, C1-C6 alkyl, C3-C6 cycloalkyl, 3-8 membered cycloheteroalkyl, C6-C10 aryl, 5-10 membered heteroaryl, -OR6, -CONR7R'7, wherein the substitution refers to the substitution of one or more R a replace:

[0181] Each R a Independently selected from: halogen, cyano, hydroxyl, C1-C6 alkyl, halogenated C1-C6 alkyl, C1-C6 alkoxy, halogenated C1-C6 alkoxy or -NR7R'7;

[0182] R6, R7 and R'7 are each independently selected from: H, C1-C6 alkyl, halogenated C1-C6 alkyl, C3-C6 cycloalkyl, halogenated C3-C6 cycloalkyl, 3-8 membered cycloheteroalkyl;

[0183] R5 is selected from the following groups selected from the group consisting of substituted or unsubstituted: -H, -CN, -CHO, nitro, R8C(O)-, R8S(O)2, -NH2, -OH, halogen, C1-C6 alkyl, 3-8 membered heteroalkyl, C1-C6 alkoxy, C3-C6 cycloalkyl, 3-8 membered heterocyclyl, C2-C6 alkenyl, C2-C6 alkynyl, -C(O)OR8, -(L1)pN(R9)(R'9), -C(O)-N(R 10 )(R' 10)、-C(O)-N(R 11 )-(L2)mN(R' 11 )(R” 11 );

[0184] or R b When R and R5 are located at two adjacent C atoms, the C atoms to which they are connected are fused to form a substituted or unsubstituted group of the following groups: C4-C8 cycloalkenyl, 4-8 membered cycloheteroyl, C6-C10 aryl, 5-10 membered heteroaryl;

[0185] R8, R 10 , R' 10 , R 11 , R' 11 and R” 11 Each independently selected from the following group of substituted or unsubstituted groups: H, C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl, 3-6 membered cycloheteroalkyl, C6-C10 aryl, 5-10 membered heteroaryl, wherein the substitution refers to substitution by one or more groups selected from the following group: halogen, hydroxyl, C1-C6 alkyl;

[0186] or R 10 and R' 10 Together with the N atom to which it is attached, it forms a substituted or unsubstituted 3-8 membered heterocyclic group;

[0187] or R' 11 and R” 11 Together with the N atom to which it is attached, it forms a substituted or unsubstituted 3-8 membered heterocyclic group;

[0188] R9 and R'9 are each independently selected from the following substituted or unsubstituted groups: H, -CN, -CHO, -NR 12 R' 12 , -OH, 3-8 membered heteroalkyl, 3-8 membered heterocyclyl, C6-C10 aryl, 5-10 membered heteroaryl, wherein R 12 and R' 12 Each is independently selected from: C1-C6 alkyl, halogenated C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl, C3-C6 cycloheteroalkyl;

[0189] or R9 and R'9 together with the N atom to which they are attached form a substituted or unsubstituted 3-8 membered heterocyclic group;

[0190] Unless otherwise specified, the substitution refers to substitution by one or more R;

[0191] Each R is independently selected from: halogen, cyano, -CHO, hydroxyl, amino, C1-C6 alkyl, C1-C6 alkoxy;

[0192] Each L1 and L2 is independently selected from: -CR f R g -, CO, SO2; among them, R f and R g Each is independently selected from the following group: H, C1-C6 alkyl, halogen, cyano, hydroxyl, amino;

[0193] r is 1, 2, or 3;

[0194] m and p are each independently 1, 2, 3, 4, 5 or 6;

[0195] n is 0, 1 or 2;

[0196] n' is 0, 1, 2 or 3;

[0197] n” is 0, 1, 2 or 3.

[0198] In another preferred embodiment, Some selected from: in, is a single bond or a double bond; q is 0, 1, 2; X1 and X2 are each independently selected from: NH, O, S or CH2; X3 and X4 are each independently selected from: N or CH; R1 is as defined above.

[0199] In another preferred embodiment, the compound of formula I or its pharmaceutically acceptable salt, hydrate, solvate, isotope compound or prodrug has a structure shown in formula I':

[0200]

[0201] In the formula,

[0202] R1 is selected from: H, halogen, hydroxy, amino, C1-C6 alkyl, halo-C1-C6 alkyl, C1-C6 alkoxy, halo-C1-C6 alkoxy, C3-C6 cycloalkyl, halo-C3-C6 cycloalkyl, C3-C6 cycloalkyloxy, halo-C3-C6 cycloalkyloxy, (C1-C6 alkyl)NH- or (C1-C6 alkyl)(C1-C6 alkyl)N-;

[0203] Each R2 is independently selected from: H, halogen, hydroxyl, amino, cyano, nitro, C1-C6 alkyl, halogenated C1-C6 alkyl, C3-C6 cycloalkyl, halogenated C3-C6 cycloalkyl;

[0204] R3 and R4 are each independently selected from: H, halogen, hydroxyl, amino, cyano, C1-C6 alkyl, halogenated C1-C6 alkyl, C3-C6 cycloalkyl, 3-8 membered cycloheteroalkyl, C6-C10 aryl, 5-10 membered heteroaryl, -OR6, -CONR7R'7, wherein the C1-C6 alkyl is optionally substituted by one or more groups selected from the group consisting of: cyano, hydroxyl, C1-C6 alkoxy, halogenated C1-C6 alkoxy and -NR7R'7, and the C6-C10 aryl or 5-10 membered heteroaryl is optionally substituted by one or more groups selected from the group consisting of: halogen, C1-C6 alkyl, halogenated C1-C6 alkyl;

[0205] R6, R7 and R'7 are each independently selected from: H, C1-C6 alkyl, halogenated C1-C6 alkyl, C3-C6 cycloalkyl, halogenated C3-C6 cycloalkyl, 3-8 membered cycloheteroalkyl;

[0206] R5 is selected from: -H, -CN, -CHO, -NH2, -OH, halogen, C1-C6 alkyl, 3-8 membered heteroalkyl, halogenated C1-C6 alkyl, C1-C6 alkyl-OH, C1-C6 alkoxy, C3-C6 cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, -C(O)OR8, -(L1)pN(R9)(R'9), -C(O)-N(R 10 )(R' 10 )、-C(O)-N(R 11 )-(L2)mN(R' 11 )(R” 11 );

[0207] R8, R 10 , R' 10 , R 11 , R' 11 and R” 11 Each is independently selected from: H, C1-C6 alkyl, halogenated C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl, 3-6 membered cycloheteroalkyl, C6-C10 aryl, 5-10 membered heteroaryl;

[0208] or R 10 and R' 10 Together with the N atom to which it is connected, it forms a substituted or unsubstituted 3-8 membered heterocyclic group, wherein the substitution refers to substitution by one or more groups selected from the group consisting of halogen, cyano, -CHO, hydroxyl, amino, C1-C6 alkyl, C1-C6 alkoxy;

[0209] R9 and R'9 are each independently selected from: -CN, -CHO, -NR 12 R' 12, -OH, 3-8 membered heteroalkyl, 3-8 membered heterocyclyl, 5-10 membered heteroaryl, wherein R 12 and R' 12 Each is independently selected from: C1-C6 alkyl, halogenated C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl, C3-C6 cycloheteroalkyl;

[0210] or R9 and R'9 together with the N atom to which they are attached form a substituted or unsubstituted 3-8 membered heterocyclic group, wherein the substitution refers to substitution by one or more groups selected from the group consisting of halogen, cyano, -CHO, hydroxyl, amino, C1-C6 alkyl, C1-C6 alkoxy;

[0211] L1 and L2 are each independently -CR f R g -, wherein the R f and R g Each is independently selected from: H, C1-C6 alkyl, halogen, cyano, hydroxyl, amino;

[0212] n is 0, 1 or 2;

[0213] m and p are each independently 1, 2, 3, 4, 5 or 6.

[0214] In another preferred embodiment, R1 is selected from: C1-C6 alkoxy, halogenated C1-C6 alkoxy, C3-C6 cycloalkoxy, halogenated C3-C6 cycloalkoxy.

[0215] In another preferred embodiment, R5 is selected from: -CN, -CHO, C1-C6 alkyl, halogenated C1-C6 alkyl, 3-8 membered heteroalkyl, C1-C6 alkyl-OH, C2-C6 alkenyl, C2-C6 alkynyl, -C(O)OR8, -(L1)pN(R9)(R'9), -C(O)-N(R 10 )(R' 10 )、-C(O)-N(R 11 )-(L2)mN(R' 11 )(R” 11 );

[0216] R8, R 10 , R' 10 , R 11 , R' 11 and R” 11 Each is independently selected from: H, C1-C6 alkyl, halogenated C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl, C3-C6 cycloheteroalkyl, C6-C10 aryl, 5-10 membered heteroaryl;

[0217] or R 10 and R'10 Together with the N atom to which it is connected, it forms a substituted or unsubstituted 3-8 membered heterocyclic group, wherein the substitution refers to substitution by one or more groups selected from the group consisting of halogen, cyano, -CHO, hydroxyl, amino, C1-C6 alkyl, C1-C6 alkoxy;

[0218] R9 and R'9 are each independently selected from: -CN, -CHO, -NR 12 R' 12 , -OH, 3-8 membered heteroalkyl, 3-8 membered heterocyclyl, 5-10 membered heteroaryl, wherein R 12 and R' 12 Each is independently selected from: C1-C6 alkyl, halogenated C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl, C3-C6 cycloheteroalkyl;

[0219] or R9 and R'9 together with the N atom to which they are attached form a substituted or unsubstituted 3-8 membered heterocyclic group, wherein the substitution refers to substitution by one or more groups selected from the group consisting of halogen, cyano, -CHO, hydroxyl, amino, C1-C6 alkyl, C1-C6 alkoxy;

[0220] L1 and L2 are each independently -CR f R g -, wherein the R f and R g Each is independently selected from: H, C1-C6 alkyl, halogen, cyano, hydroxyl, amino;

[0221] m and p are each independently 1, 2, 3, 4, 5 or 6.

[0222] In another preferred embodiment, "-(L1) p -" and "-(L2) m -" is -CH2CH2-.

[0223] In another preferred embodiment, m is 1, 2, 3 or 4.

[0224] In another preferred embodiment, p is 1, 2, 3 or 4.

[0225] In another preferred embodiment, each R2 is independently selected from: halogen, cyano, nitro, C1-C6 alkyl, halogenated C1-C6 alkyl.

[0226] In another preferred embodiment, R3 and R4 are each independently selected from: H, halogen, hydroxyl, amino, cyano, C1-C6 alkyl, halogenated C1-C6 alkyl.

[0227] In another preferred embodiment, the compound of formula I has a structure shown in formula I":

[0228]

[0229] In the formula,

[0230] Rc is selected from substituted or unsubstituted C1-C6 alkyl, wherein the substitution refers to substitution by one or more halogen, cyano, or hydroxyl groups;

[0231] X is selected from: halogen, cyano, nitro, halogenated C1-C6 alkyl;

[0232] R5 is as defined above.

[0233] In another preferred embodiment, R5 is selected from:

[0234] In another preferred embodiment, R5 is -C(O)OR8, wherein R8 is selected from: methyl, ethyl, isopropyl, and tert-butyl.

[0235] In another preferred embodiment, R5 is -(L1)pN(R9)(R'9), wherein p is 1, 2, or 3; and R9 and R'9 are as defined above.

[0236] In another preferred embodiment, R5 is -C(O)-N(R 10 )(R' 10 ), where R 10 and R' 10 Each is independently selected from: H, methyl, ethyl, isopropyl, tert-butyl, C3-C6 cycloalkyl (such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl), 3-6 membered heterocycloalkane (such as tetrahydrofuranyl, tetrahydropyrrolyl, morpholinyl), or R 10 and R' 10 Together with the nitrogen atom to which it is attached, it forms a 5-6 membered heterocyclic group.

[0237] In another preferred embodiment, R5 is -C(O)-NH-CH2CH2-N(R' 11 )(R” 11 ), where R' 11 and R” 11 Each is independently selected from: H, methyl, ethyl, isopropyl, tert-butyl, C3-C6 cycloalkyl (such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl), 3-6 membered heterocycloalkane (such as tetrahydrofuranyl, tetrahydropyrrolyl, morpholinyl).

[0238] In another preferred embodiment, R1, R2, R3, R4, and R5 are specific groups corresponding to the specific compounds in the examples.

[0239] The salts that may be formed by the compounds of the present invention also belong to the scope of the present invention. Unless otherwise specified, the compounds of the present invention are understood to include their salts. The term "salt" used herein refers to an acidic or basic salt formed with an inorganic or organic acid and a base. In addition, when the compound of the present invention contains a basic fragment, it includes but is not limited to pyridine or imidazole, and contains an acidic fragment, including but not limited to carboxylic acid, the zwitterions ("inner salts") that may be formed are included in the scope of the term "salt". Pharmaceutically acceptable (i.e., non-toxic, physiologically acceptable) salts are preferred, although other salts are also useful, for example, in separation or purification steps in the preparation process. The compounds of the present invention may form salts, for example, Compound I reacts with a certain amount of acid or base, such as an equivalent amount, and is salted out in a medium, or freeze-dried in an aqueous solution.

[0240] The compounds of the present invention contain basic fragments, including but not limited to amines or pyridine or imidazole rings, which may form salts with organic or inorganic acids. Typical acids that can form salts include acetates (such as acetic acid or trihaloacetic acid, such as trifluoroacetic acid), adipates, alginate, ascorbate, aspartate, benzoate, benzenesulfonate, bisulfate, borate, butyrate, citrate, camphor salt, camphorsulfonate, cyclopentanepropionate, diglycolate, dodecyl sulfate, ethanesulfonate, fumarate, glucoheptonate, glycerophosphate, hemisulfate, heptanoate, hexanoate, hydrochloride, hydrobromide, hydroiodide

[0013] The invention also includes but is not limited to the following: esters, isothioates (e.g., 2-hydroxyethanesulfonate), lactates, maleates, methanesulfonates, naphthalenesulfonates (e.g., 2-naphthalenesulfonate), nicotinates, nitrates, oxalates, pectinates, persulfates, phenylpropionates (e.g., 3-phenylpropionate), phosphates, picrates, pivalates, propionates, salicylates, succinates, sulfates (e.g., formed with sulfuric acid), sulfonates, tartrates, thiocyanates, toluenesulfonates such as p-toluenesulfonates, dodecanoates, and the like.

[0241] Certain compounds of the present invention may contain acidic fragments, including but not limited to carboxylic acids, which may form salts with various organic or inorganic bases. Typical base-formed salts include ammonium salts, alkali metal salts such as sodium, lithium, potassium salts, alkaline earth metal salts such as calcium, magnesium salts and salts formed with organic bases (such as organic amines), such as benzathine, dicyclohexylamine, hepamine (salt formed with N,N-di(dehydroabietyl)ethylenediamine), N-methyl-D-glucamine, N-methyl-D-glucamide, tert-butylamine, and salts formed with amino acids such as arginine, lysine, etc. Basic nitrogen-containing groups can be combined with halide quaternary ammonium salts, such as small molecule alkyl halides (such as chlorides, bromides and iodides of methyl, ethyl, propyl and butyl), dialkyl sulfates (such as dimethyl sulfate, diethyl sulfate, dibutyl sulfate and dipentyl sulfate), long chain halides (such as chlorides, bromides and iodides of decyl, dodecyl, tetradecyl and tetradecyl), aralkyl halides (such as benzyl and phenyl bromides), etc.

[0242] As used herein, the term "pharmaceutically acceptable salt" refers to a salt formed by a compound of the present invention and an acid or base that is suitable for use as a drug. Pharmaceutically acceptable salts include inorganic salts and organic salts. A preferred class of salts is a salt formed by a compound of the present invention and an acid. Suitable acids for forming salts include, but are not limited to, inorganic acids such as hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, nitric acid, and phosphoric acid; organic acids such as formic acid, acetic acid, trifluoroacetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, picric acid, benzoic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, benzenesulfonic acid, and naphthalenesulfonic acid; and amino acids such as proline, phenylalanine, aspartic acid, and glutamic acid.

[0243] Another preferred salt is a salt of the compound of the present invention and a base, such as an alkali metal salt (e.g., sodium salt or potassium salt), an alkaline earth metal salt (e.g., magnesium salt or calcium salt), an ammonium salt (e.g., lower alkanolammonium salt and other pharmaceutically acceptable amine salts), for example, methylamine salt, ethylamine salt, propylamine salt, dimethylamine salt, trimethylamine salt, diethylamine salt, triethylamine salt, tert-butylamine salt, ethylenediamine salt, hydroxyethylamine salt, dihydroxyethylamine salt, trihydroxyethylamine salt, and amine salts formed from morpholine, piperazine, and lysine, respectively.

[0244] The term "solvate" refers to a complex formed by the coordination of the compound of the present invention with solvent molecules in a specific ratio. "Hydrate" refers to a complex formed by the coordination of the compound of the present invention with water.

[0245] Prodrugs and solvates of the compounds of the present invention are also within the scope of the invention. The term "prodrug" herein refers to a compound that undergoes chemical transformation by metabolic or chemical processes to produce a compound, salt, or solvate of the present invention when treating a related disease. The compounds of the present invention include solvates, such as hydrates.

[0246] The compounds, salts or solvates of the present invention may exist in tautomeric forms (such as amides and imino ethers). All these tautomers are part of the present invention.

[0247] All stereoisomers of the compounds (e.g., those due to asymmetric carbon atoms that may exist for various substitutions), including enantiomeric and diastereomeric forms, are contemplated by the present invention. Individual stereoisomers of the compounds of the present invention may not exist simultaneously with other isomers (e.g., as a pure or substantially pure optical isomer having a particular activity), or may be mixtures, such as racemates, or mixtures with all other stereoisomers or portions thereof. The chiral centers of the present invention have two configurations, S or R, as defined by the 1974 recommendations of the International Union of Pure and Applied Chemistry (IUPAC). Racemic forms can be resolved by physical methods, such as fractional crystallization, or by crystallization of diastereoisomers derived therefrom, or by separation by chiral column chromatography. Individual optical isomers can be obtained from racemates by suitable methods, including but not limited to conventional methods, such as recrystallization after salt formation with an optically active acid.

[0248] The compounds of the present invention are prepared, separated and purified to obtain compounds with a weight content of equal to or greater than 90%, for example, equal to or greater than 95%, equal to or greater than 99% ("very pure" compounds), which are listed in the text description. Such "very pure" compounds of the present invention are also considered part of the present invention.

[0249] All configurational isomers of the compounds of the present invention are included in the scope, whether in mixture, pure or very pure form. The definition of the compounds of the present invention includes both cis (Z) and trans (E) olefin isomers, as well as cis and trans isomers of carbocyclic and heterocyclic rings.

[0250] Throughout the specification, groups and substituents may be selected to provide stable fragments and compounds.

[0251] Specific functional groups and chemical term definitions are detailed below. For purposes of this invention, chemical elements are referred to in the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 75 th Ed. Definitions of specific functional groups are also described therein. In addition, the basic principles of organic chemistry as well as specific functional groups and reactivity are also described in "Organic Chemistry", Thomas Sorrell, University Science Books, Sausalito: 1999, which is incorporated by reference in its entirety.

[0252] Certain compounds of the present invention may exist in specific geometric or stereoisomeric forms. The present invention encompasses all compounds, including their cis and trans isomers, R and S enantiomers, diastereomers, (D) isomers, (L) isomers, racemic mixtures and other mixtures. In addition, asymmetric carbon atoms may represent substituents, such as alkyl. All isomers and their mixtures are included in the present invention.

[0253] According to the present invention, the ratio of isomers contained in the mixture of isomers can be various. For example, in the mixture of only two isomers, there can be the following combinations: 50:50, 60:40, 70:30, 80:20, 90:10, 95:5, 96:4, 97:3, 98:2, 99:1, or 100:0, and all ratios of isomers are within the scope of the present invention. Similar ratios that are easily understood by ordinary technicians in this profession, and ratios for more complex mixtures of isomers are also within the scope of the present invention.

[0254] The present invention also includes isotopically labeled compounds that are equivalent to the original compounds disclosed herein. However, in practice, it is common for one or more atoms to be replaced by atoms having a different atomic mass or mass number than the original atoms. Examples of isotopes of the compounds of the present invention include hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine and chlorine isotopes, such as 2 H. 3 H. 13 C. 11 C. 14 C. 15 N. 18 O. 17 O. 31 P. 32 P. 35 S. 18 F and 36 The compounds of the present invention, or enantiomers, diastereomers, isomers, or pharmaceutically acceptable salts or solvates, wherein the isotopes or other isotopic atoms of the above compounds are all within the scope of the present invention. Certain isotope-labeled compounds of the present invention, such as 3 H and 14 Radioisotopes of C are also included and are useful in tissue distribution experiments of drugs and substrates. 3 H and carbon-14, i.e. 14 C, their preparation and detection are relatively easy. It is the first choice among isotopes. In addition, heavier isotope substitutions such as deuterium, i.e. 2H, due to its good metabolic stability in certain therapies has advantages, such as increasing half-life or reducing dosage in vivo, therefore, in some cases, it can be preferred. Isotope-labeled compounds can be prepared by general methods by replacing readily available isotope-labeled reagents with non-isotopic reagents using the schemes disclosed in the examples.

[0255] If a synthesis of a specific enantiomer of the compound of the present invention is to be designed, it can be prepared by asymmetric synthesis, or derivatized with a chiral auxiliary, the resulting diastereomeric mixture can be separated, and the chiral auxiliary can be removed to obtain the pure enantiomer. In addition, if the molecule contains a basic functional group, such as an amino acid, or an acidic functional group, such as a carboxyl group, a diastereomeric salt can be formed with a suitable optically active acid or base, and then separated by conventional means such as fractional crystallization or chromatography, and then the pure enantiomer can be obtained.

[0256] As described herein, the compounds of the present invention can be replaced with any number of substituents or functional groups to expand their scope. Generally, the term "substituted" appears before or after the term "optional", and the general formula including substituents in the formulation of the present invention refers to replacing hydrogen free radicals with designated structural substituents. When multiple positions in a specific structure are substituted by multiple specific substituents, the substituents can be the same or different at each position. The term "substituted" used herein includes all allowed organic compound substitutions. In a broad sense, allowed substituents include non-cyclic, cyclic, branched non-branched, carbocyclic and heterocyclic, aromatic and non-aromatic organic compounds. In the present invention, heteroatom nitrogen can have hydrogen substituents or any allowed organic compounds described above to supplement its valence. In addition, the present invention is not intended to limit the allowed substitution of organic compounds in any way. The present invention believes that the combination of substituents and variable groups is very good in the treatment of diseases in the form of stable compounds. As used herein, the term "stable" refers to compounds that are stable and that maintain the structural integrity of the compound over a period of time sufficient to be detected, and preferably over a period of time sufficient to be effective, as used herein for the purposes described above.

[0257] The metabolites of the compounds and pharmaceutically acceptable salts thereof involved in the present application, as well as prodrugs that can be converted into the structures of the compounds and pharmaceutically acceptable salts involved in the present application and in vivo, are also included in the claims of the present application.

[0258] Pharmaceutical compositions and methods of administration

[0259] The pharmaceutical composition of the present invention is used for preventing and / or treating the following diseases: irritable bowel syndrome and / or cancer.

[0260] The compounds of general formula (I) can be used in combination with other drugs known to treat or improve similar conditions. When administered in combination, the administration method and dosage of the original drug can remain unchanged, and the compound of formula I can be taken simultaneously or subsequently. When the compound of formula I is taken simultaneously with one or more other drugs, a pharmaceutical composition containing one or more known drugs and the compound of formula I can be preferably used. Drug combination also includes taking the compound of formula I and one or more other known drugs in overlapping time periods. When the compound of formula I is used in combination with one or more other drugs, the dosage of the compound of formula I or the known drug may be lower than the dosage of them taken alone.

[0261] The drugs or active ingredients that can be used in combination with the compound of formula (I) include but are not limited to: aminosalicylic acid preparations (such as sulfasalazine), glucocorticoids (such as hydrocortisone, dexamethasone, etc.), PD-1 inhibitors (such as nivolumab, pembrolizumab, JS-001, SHR-120, BGB-A317, IBI-308, GLS-010, GB-226, STW204, HX008, HLX10, BAT1306, AK105, LZM009 or biosimilars of the above drugs, etc.), PD-L1 inhibitors (such as durvalumab, atezolizumab, CS1001, KN035, HLX20, SHR-1316, BGB-A333, JS003, CS1003, KL-A167, F520, GR1405, MSB2311 or biosimilars of the above drugs, etc.), CD20 antibodies (such as rituximab, obinutuzumab, ofatumumab, tositumomab, ibritumomab tiuxetan, etc.), CD47 antibodies (such as Hu5F9-G4, CC-90002, TTI-621, TTI-622 , OSE-172, SRF-231, ALX-148, NI-1701, SHR-1603, IBI188, IMM01), ALK inhibitors (such as Ceritinib, Alectinib, Brigatinib, Lorlatinib, Oclatinib), PI3K inhibitors (such as Idelalisib, Dactolisib, Taselisib, Buparlisib, etc.), BTK inhibitors (such as Ibrutinib, Tirabrutinib, Acalabrutinib, etc.), EGFR inhibitors (such as Afatinib, Gefitinib, Erlotinib, Lapatinib, Dacomitinib, etc.), , icotinib, canertinib, etc.), VEGFR inhibitors (such as sorafenib, pazopanib, rivatinib, cabozantinib, sunitinib, donafenib, etc.), HDAC inhibitors (such as givinostat, droxinostat, entinostat, dacilast, tekdinib, etc.), CDK inhibitors (such as palbociclib, ribociclib, abemaciclib, lerociclib, etc.), MEK inhibitors (such as selumetinib (AZD6244), trametinib (GSK1120212), PD0325901, U0126, AS-703026, P D184352 (CI-1040), etc.), Akt inhibitors (such as MK-2206, Ipatasertib, Capivasertib, Afuresertib, Uprosertib, etc.), mTOR inhibitors (such as Vistusertib, etc.), SHP2 inhibitors (such as RMC-4630, JAB-3068, TNO155, etc.), IGF-1R inhibitors (such as Ceritinib, Ocasatinib, linsitinib, BMS-754807, GSK1838705A, etc.) or any combination thereof.

[0262] The dosage forms of the pharmaceutical composition of the present invention include (but are not limited to): injection, tablet, capsule, aerosol, suppository, film, pill, external ointment, controlled release, sustained release or nano preparation.

[0263] The pharmaceutical composition of the present invention comprises a safe and effective amount of the compound of the present invention or a pharmacologically acceptable salt thereof and a pharmacologically acceptable excipient or carrier. Wherein "safe and effective amount" means: the amount of the compound is sufficient to significantly improve the condition without causing serious side effects. Usually, the pharmaceutical composition contains 1-2000 mg of the compound of the present invention per dose, and more preferably, contains 10-1000 mg of the compound of the present invention per dose. Preferably, the "one dose" is a capsule or tablet.

[0264] "Pharmaceutically acceptable carrier" refers to: one or more compatible solid or liquid fillers or gel substances, which are suitable for human use and must have sufficient purity and sufficiently low toxicity. "Compatibility" here means that the components in the composition can be mixed with the compounds of the present invention and with each other without significantly reducing the efficacy of the compounds. Some examples of pharmaceutically acceptable carriers include cellulose and its derivatives (such as sodium carboxymethyl cellulose, sodium ethyl cellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerol, mannitol, sorbitol, etc.), emulsifiers (such as ), wetting agents (such as sodium lauryl sulfate), colorants, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

[0265] There is no particular limitation on the administration of the compound or pharmaceutical composition of the present invention. Representative administrations include, but are not limited to, oral, intratumoral, rectal, parenteral (intravenous, intramuscular or subcutaneous), and topical administration.

[0266] Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with the following ingredients: (a) fillers or extenders, for example, starches, lactose, sucrose, glucose, mannitol, and silicic acid; (b) binders, for example, hydroxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose, and acacia; (c) humectants, for example, glycerol; (d) disintegrants, for example, agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) solubilizers, for example, paraffin; (f) absorption accelerators, for example, quaternary ammonium compounds; (g) wetting agents, for example, cetyl alcohol and glyceryl monostearate; (h) adsorbents, for example, kaolin; and (i) lubricants, for example, talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, or mixtures thereof. In the case of capsules, tablets and pills, the dosage forms may also comprise buffering agents.

[0267] Solid dosage forms such as tablets, pills, capsules, pills and granules can be prepared using coatings and shell materials, such as enteric coatings and other materials known in the art. They may contain opacifiers, and the release of the active compound or compounds in such compositions can be delayed in a certain part of the digestive tract. Examples of embedding components that can be used are polymeric substances and waxes. If necessary, the active compound can also be formed into microencapsulated form with one or more of the above-mentioned excipients.

[0268] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the active compound, the liquid dosage form may contain an inert diluent conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butylene glycol, dimethylformamide and oils, in particular cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil or mixtures of these substances.

[0269] Besides such inert diluents, the composition may also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

[0270] Suspensions, in addition to the active compounds, may contain suspending agents such as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methanol and agar, or mixtures of these substances, and the like.

[0271] Compositions for parenteral injection may include physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions or emulsions and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and non-aqueous carriers, diluents, solvents or excipients include water, ethanol, polyols and suitable mixtures thereof.

[0272] Dosage forms for topical administration of the compounds of the invention include ointments, powders, patches, sprays and inhalants. The active ingredient is mixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or propellants that may be required.

[0273] The treatment method of the present invention can be used alone or in combination with other treatment methods or therapeutic drugs.

[0274] When using the pharmaceutical composition, a safe and effective amount of the compound of the present invention is applied to a mammal (such as a human) in need of treatment, wherein the dosage during administration is a pharmaceutically effective dosage, and for a person weighing 60 kg, the daily dosage is usually 1 to 2000 mg, preferably 50 to 1000 mg. Of course, the specific dosage should also take into account factors such as the route of administration and the health status of the patient, which are all within the skill of a skilled physician.

[0275] The present invention also provides a method for preparing a pharmaceutical composition, comprising the steps of: mixing a pharmaceutically acceptable carrier with the compound of general formula (I) of the present invention or its crystal form, pharmaceutically acceptable salt, hydrate or solvate to form a pharmaceutical composition.

[0276] The present invention also provides a treatment method, which comprises the steps of administering to a subject in need of treatment a compound of formula (I) of the present invention, or a crystalline form, a pharmaceutically acceptable salt, a hydrate or a solvate thereof, or administering a pharmaceutical composition of the present invention, for selectively inhibiting RET.

[0277] Preparation method

[0278] Methods for preparing compounds of Formula I are described in the following schemes and examples. Starting materials and intermediates were purchased from commercial sources, prepared by known procedures, or otherwise described. In some cases, the order in which the steps of the reaction schemes are carried out may be altered to facilitate the reaction or to avoid unwanted side reaction products.

[0279] The preparation method of the compound of formula I of the present invention is described in more detail below, but these specific methods do not constitute any limitation to the present invention. The compounds of the present invention can also be conveniently prepared by optionally combining various synthetic methods described in this specification or known in the art, and such a combination can be easily carried out by a technician in the field to which the present invention belongs.

[0280] Usually, in the preparation process, each reaction is usually carried out under the protection of inert gas in a suitable solvent at 0 to 90°C, and the reaction time is usually 2 to 24 hours.

[0281] Preferably, the preparation method of the compound of formula I of the present invention comprises the following steps:

[0282] (s1) Compound 1-1 is slowly added with concentrated nitric acid or fuming nitric acid in the presence of concentrated sulfuric acid to carry out a nitration reaction to obtain compound 1-2;

[0283] (s2) Compound 1-2 is subjected to functional group conversion and reduction steps to obtain compound I1;

[0284] (s3) Compound 2-1 reacts in an inert solvent under alkaline conditions to generate compound 2-2;

[0285] (s4) Compound 2-2 is subjected to coupling reaction to generate compound I2

[0286] (s5) reacting the compound of formula I1 and the compound of formula I2 in an inert solvent in the presence of a catalyst to obtain a compound of formula 3-1;

[0287] (s6) reacting the compound of formula 3-1 in an inert solvent under acidic conditions to obtain a compound of formula I;

[0288]

[0289] In the formula, Y is selected from OH, halogen;

[0290] R1, R2, R5, R b , r, n, n' and n" are as defined above.

[0291] Preferably, in steps (s3), (s5) and (s6), the inert solvent is pyridine, DMF, DMSO, triethylamine, DCM, 1,2-dichloroethane, tetrahydrofuran or 1,6-dioxane.

[0292] Preferably, in step (s5), the catalyst is propyl phosphoric anhydride (T3P).

[0293] Preferably, in step (s6), the acid is TFA.

[0294] Preferably, the compound of the present invention can be obtained by the following steps:

[0295]

[0296] Wherein, R1 and R5 are as defined above;

[0297] Preferably, R1 is

[0298] Preferably, R5 is selected from

[0299] (i) slowly adding concentrated nitric acid or fuming nitric acid dropwise in the presence of concentrated sulfuric acid to carry out a nitration reaction to obtain compound 2;

[0300] (ii) the acid is converted into an acyl chloride in the presence of thionyl chloride or oxalyl chloride and a catalyst (e.g., DMF). Under alkaline conditions (e.g., Et3N, DIEA), the acyl chloride and amine react rapidly to form compound 3;

[0301] (iii) under acidic conditions (e.g., HCl), iron reduces the nitro group to obtain compound 4;

[0302] (iv) under alkaline conditions (such as potassium carbonate), compound 5 reacts with an iodoalkyl to produce compound 6;

[0303] (v) under alkaline conditions (such as potassium hydroxide), compound 6 reacts with p-methoxyphenylmethanol to generate compound 7;

[0304] (vi) under alkaline conditions (such as cesium carbonate), compound 7 is coupled with methyl 2-(2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)acetate to generate compound 8;

[0305] (vii) under alkaline conditions (such as lithium hydroxide), compound 8 is demethylated to generate compound 9;

[0306] (viii) Compound 4 and compound 9 are condensed into amide under alkaline (e.g., pyridine) and acid anhydride conditions to generate compound 10;

[0307] (ix) Compound 10 is subjected to the removal of the benzyloxy group under acidic conditions (e.g., TFA) to obtain compound 11;

[0308] In the above reaction steps, the reaction solvent, reaction temperature, reaction time, catalyst, etc. can be selected according to the specific reactants.

[0309] In the above reaction steps, the reaction raw materials and reagents can be purchased commercially or synthesized via routes reported in literature.

[0310] The present invention has the following main advantages:

[0311] (1) The compounds of the present invention have excellent inhibitory ability and selectivity for RET kinase, and have low inhibitory activity against other kinases such as VEGFR2;

[0312] (2) The compounds of the present invention have excellent inhibitory ability against TRK kinases, and have stronger inhibitory ability against mutant TRK kinases;

[0313] (3) Some of the compounds in the present invention have better cell inhibitory activity and show better pharmacodynamic effects in the IBS model (acetic acid-induced intestinal hypersensitivity model).

[0314] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention. The experimental methods in the following examples without specifying specific conditions are usually based on conventional conditions or the conditions recommended by the manufacturer. Unless otherwise stated, percentages and parts are calculated by weight.

[0315] Unless otherwise defined, all professional and scientific terms used herein have the same meanings as those familiar to those skilled in the art. In addition, any methods and materials similar or equivalent to those described herein can be applied to the methods of the present invention. The preferred implementation methods and materials described herein are for demonstration purposes only.

[0316] The structure of the compound of the present invention is confirmed by nuclear magnetic resonance (NMR) and liquid chromatography-mass spectrometry (LC-MS).

[0317] NMR was detected using Bruker AVANCE-400 and Bruker AVANCE-500 nuclear magnetic spectrometers. The measurement solvents included deuterated dimethyl sulfoxide (DMSO-d6), deuterated acetone (CD3COCD3), deuterated chloroform (CDCl3) and deuterated methanol (CD3OD), etc. Tetramethylsilane (TMS) was used as the internal standard, and the chemical shift was measured in parts per million (ppm).

[0318] Liquid chromatography-mass spectrometry (LC-MS) was detected using an Agilent 1260 mass spectrometer. HPLC was determined using an Agilent 1100 high pressure chromatograph (Microsorb 5micron C18 100 x 3.0 mm m, and the preparative thin layer chromatography used was 0.4 mm-0.5 mm. Column chromatography generally used Qingdao silica gel 200-300 mesh silica gel as a carrier.

[0319] The starting materials in the examples of the present invention are all known and commercially available, or can be synthesized using or according to literature data reported in the art.

[0320] Unless otherwise specified, all reactions of the present invention are carried out under the protection of dry inert gas (such as nitrogen or argon) with continuous magnetic stirring, and the reaction temperatures are all degrees Celsius.

[0321] The following abbreviations are used throughout this invention:

[0322] THF: Tetrahydrofuran

[0323] DCM: dichloromethane

[0324] Fe:Iron powder

[0325] H2SO4: sulfuric acid

[0326] HNO3:nitric acid

[0327] SOCl2: Sulfonyl chloride

[0328] HCl: hydrochloric acid

[0329] H2O: water

[0330] TEA: triethylamine

[0331] DIEA: N,N-diisopropylethylamine

[0332] DMF: N,N-dimethylformamide

[0333] Cs2CO3: Cesium carbonate

[0334] LiOH: lithium hydroxide

[0335] EA: Ethyl acetate

[0336] Pd(dppf)2Cl2:[1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride

[0337] T3P: Propylphosphonic anhydride

[0338] Intermediate 1 Synthesis of 3-nitro-5-(pentafluorothio)benzoic acid

[0339]

[0340] 3-(Pentafluorothio)benzoic acid (1.0 g, 4.03 mmol) was added to a single-mouth bottle, and then concentrated sulfuric acid (12 mL) was slowly added. The mixture was stirred and cooled to 0°C, and concentrated nitric acid (2 mL) was slowly added. The mixture was stirred for 10 minutes, and then the temperature was raised to 80°C for reaction for 12 hours. After the reaction was complete, the mixture was cooled and slowly poured into ice water. The mixture was extracted with EA, and the organic phase was neutralized with saturated sodium bicarbonate, dried, filtered, and concentrated to obtain 0.9 g of 3-nitro-5-(pentafluorothio)benzoic acid.

[0341] Intermediate 2 Synthesis of 3-amino-N-(2-(dimethylamino)ethyl)-5-(pentafluorothio)benzamide

[0342]

[0343] Step 1 Synthesis of N-(2-(dimethylamino)ethyl)-3-nitro-5-(pentafluorothio)benzamide

[0344] 3-Nitro-5-(pentafluorothio)benzoic acid (0.5 g, 1.71 mmol) was dissolved in SOCl2 (10 mL), and a few drops of DMF were added. The mixture was refluxed for 2 hours. After the reaction was complete, the reaction solution was dried by spin drying. The acid chloride was dissolved in DCM (15 mL). The mixture was cooled to 0°C, and triethylamine (0.52 g, 5.12 mmol) was added. Then, N,N-dimethylethylenediamine (0.23 g, 2.56 mmol) was added. The reaction was continued at 0°C for half an hour. After the reaction was complete, water was added to quench the reaction, the liquid was separated, the organic phase was concentrated, and column chromatography was performed to obtain 0.55 g of N-(2-(dimethylamino)ethyl)-3-nitro-5-(pentafluorothio)benzamide. MS m / z (ESI): 363.3 [M+H] + .

[0345] Step 2 Synthesis of 3-amino-N-(2-(dimethylamino)ethyl)-5-(pentafluorothio)benzamide

[0346] N-(2-(dimethylamino)ethyl)-3-nitro-5-(pentafluorothio)benzamide (0.5 g, 1.38 mmol) was added to methanol (10 mL), and then iron powder (1.5 g) was added, and then hydrochloric acid (1 mL, 12 mol / L) was slowly added. The mixture was stirred at 72°C for 2 hours. After the reaction was completed, it was cooled and filtered. The reaction solution was spin-dried and column chromatography was performed to obtain 0.38 g of 3-amino-N-(2-(dimethylamino)ethyl)-5-(pentafluorothio)benzamide. MS m / z (ESI): 333.9 [M+H] + .

[0347] Intermediate 3 Synthesis of 2-(4(4-ethoxy-6-((4-methoxybenzyl)oxy)pyridin-3-yl)-2-fluorophenyl)acetic acid

[0348]

[0349] Step 1 Synthesis of 5-bromo-4-ethoxy-2((4-methoxybenzyl)oxy)pyridine

[0350] At room temperature, (4-methoxyphenyl)methanol (1.75 g, 12.7 mmol), KOH (1.2 g, 21.2 mmol) and 18-crown-6 (0.28 g, 1.06 mmol) were added to a mixture of 5-bromo-2-chloro-4-ethoxypyridine (2.5 g, 10.6 mmol) in toluene (30 mL). The mixture was reacted at 120°C for 3 hours. After the reaction was complete, the reaction solution was dried by spin drying, water (15 mL) was added, and then extracted with EA (30 mL*2). The organic phase was dried, dried by spin drying, and column chromatography was performed to obtain 3.0 g of 5-bromo-4-ethoxy-2((4-methoxybenzyl)oxy)pyridine. MS m / z (ESI): 339.1 [M+H] + .

[0351] Step 2 Synthesis of methyl 2-(4(4-ethoxy-6-((4-methoxybenzyl)oxy)pyridin-3-yl)-2-fluorophenyl)acetate

[0352] Under nitrogen protection, 5-bromo-4-ethoxy-2((4-methoxybenzyl)oxy)pyridine (1.9 g, 5.62 mmol), methyl 2-(2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)acetate (1.82 g, 6.18 mmol), PdCl2(dppf) (0.45 g, 0.56 mmol) and cesium carbonate (3.65 g, 11.2 mmol) were added to a single-necked bottle, and 1,4-dioxane (30 mL) and water (10 mL) were added, and the mixture was reacted at 95 ° C for 3 hours. After the reaction was complete, the mixture was extracted with EA (40 mL*2), the organic phase was dried, spin-dried, and column chromatography was performed to obtain 1.8 g of methyl 2-(4(4-ethoxy-6-((4-methoxybenzyl)oxy)pyridin-3-yl)-2-fluorophenyl)acetate. MS m / z (ESI): 426.3 [M+H] + .

[0353] Step 3 Synthesis of 2-(4(4-ethoxy-6-((4-methoxybenzyl)oxy)pyridin-3-yl)-2-fluorophenyl)acetic acid

[0354] 2-(4(4-ethoxy-6-((4-methoxybenzyl)oxy)pyridin-3-yl)-2-fluorophenyl)acetic acid methyl ester (2.4g, 5.63mmol) was dissolved in THF (15mL), and then an aqueous solution of lithium hydroxide (0.3g, 12.4mmol) was added thereto, and the mixture was stirred at 60°C for 4 hours. After the reaction was complete, most of the organic phase was dried, and then the pH was adjusted to 7 with dilute hydrochloric acid to precipitate a white solid, which was filtered, and the filter cake was washed with water and dried to obtain 2.1g of 2-(4(4-ethoxy-6-((4-methoxybenzyl)oxy)pyridin-3-yl)-2-fluorophenyl)acetic acid, MS m / z (ESI): 412.2 [M+H] + .

[0355] Example 1 Synthesis of N-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl-2-fluorophenyl)acetylamino-5-(pentafluorothio)benzamide

[0356]

[0357] Step 1 Synthesis of N-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-((4-methoxybenzyl)oxy)pyridin-3-yl)-2-fluorophenyl)acetamido)-5-(pentafluorothio)benzamide

[0358] 2-(4(4-ethoxy-6-((4-methoxybenzyl)oxy)pyridin-3-yl)-2-fluorophenyl)acetic acid (0.6 g, 1.45 mmol) was added to pyridine (5 mL), and then 3-amino-N-(2-(dimethylamino)ethyl)-5-(pentafluorothio)benzamide (0.48 g, 1.75 mmol) and T3P (0.55 g, 1.75 mmol) were added, and the mixture was reacted at room temperature for 12 hours. After the reaction was complete, the reaction solution was directly spin-dried, and then column chromatography was performed to obtain 0.82 g of N-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-((4-methoxybenzyl)oxy)pyridin-3-yl)-2-fluorophenyl)acetamido)-5-(pentafluorothio)benzamide, MS m / z (ESI): 727.5 [M+H] + .

[0359] Step 2 Synthesis of N-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl-2-fluorophenyl)acetylamino-5-(pentafluorothio)benzamide

[0360] N-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-((4-methoxybenzyl)oxy)pyridin-3-yl)-2-fluorophenyl)acetamido)-5-(pentafluorothio)benzamide (0.82 g, 1.13 mmol) was dissolved in DCM (10 mL), and then TFA (2 mL) was added, and the mixture was stirred at room temperature for 2 hours. After the reaction was complete, the reaction solution was concentrated, and then adjusted to neutral with sodium carbonate aqueous solution, extracted with DCM (20 mL*2), and the organic phase was dried by spin drying. Column chromatography gave 0.48 g of N-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl-2-fluorophenyl)acetamido-5-(pentafluorothio)benzamide (Compound 1). MS m / z (ESI): 607.3 [M+H] + . 1 H NMR(400MHz,DMSO)δ11.38(s,1H),10.85(s,1H),8.78(s,1H),8.45(s,1H),8.24(s,1H),8.01(s,1H),7.40–7.30(m,2H),7.28– 7.17(m,2H),5.80(s,1H),4.02(dt,J=6.8,2.1Hz,4H),3.78(s,2H),2.43(t,J=6.5Hz,2H),2.19(s,6H),1.27(t,J=6.9Hz,3H).

[0361] Example 2 Synthesis of 3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-N,N-dimethyl-5-(pentafluorothio)benzamide

[0362]

[0363] Step 1 Synthesis of N,N-dimethyl-3-nitro-5-(pentafluorothio)benzamide

[0364] Dissolve 3-nitro-5-(pentafluorothio)benzoic acid (0.3 g, 1.02 mmol) in SOCl2 (10 mL), add a few drops of DMF, and then reflux for 2 hours. After the reaction is complete, spin dry the reaction solution, dissolve the acid chloride in DCM (10 mL), cool the mixture to 0°C, then add triethylamine (0.32 g, 3.07 mmol), followed by dimethylamine (0.07 g, 1.56 mmol), and continue to react at 0°C for half an hour. After the reaction is complete, add water to quench the reaction, separate the liquids, concentrate the organic phase, and column chromatography to obtain 0.26 g of N, N-dimethyl-3-nitro-5-(pentafluorothio)benzamide. MS m / z (ESI): 321.5 [M+H] + .

[0365] Step 2 Synthesis of 3-amino-N,N-dimethyl-5-(pentafluorothio)benzamide

[0366] Add N, N-dimethyl-3-nitro-5-(pentafluorothio)benzamide (0.26 g, 0.8 mmol) to methanol (10 mL), then add iron powder (0.8 g), and then slowly add hydrochloric acid (1 mL, 12 mol / L), and the mixture is stirred at 72°C for 2 hours. After monitoring the reaction is complete, cool, filter, spin dry the reaction solution, and column chromatography to obtain 0.17 g of 3-amino-N, N-dimethyl-5-(pentafluorothio)benzamide. MS m / z (ESI): 291.1 [M+H] + .

[0367] Step 3 Synthesis of 3-(2-(4-(4-ethoxy-6-((4-methoxybenzyl)oxy)pyridin-3-yl)-2-fluorophenyl)acetamido)-N,N-dimethyl-5-(pentafluorothio)benzamide

[0368] 2-(4(4-ethoxy-6-((4-methoxybenzyl)oxy)pyridin-3-yl)-2-fluorophenyl)acetic acid (0.2 g, 0.49 mmol) was added to pyridine (5 mL), and then 3-amino-N, N-dimethyl-5-(pentafluorothio)benzamide (0.17 g, 0.58 mmol) and T3P (0.19 g, 0.58 mmol) were added, and the mixture was reacted at room temperature for 12 hours. After the reaction was complete, the reaction solution was directly spin-dried, and then column chromatography was performed to obtain 3-(2-(4-(4-ethoxy-6-((4-methoxybenzyl)oxy)pyridin-3-yl)-2-fluorophenyl)acetamido)-N, N-dimethyl-5-(pentafluorothio)benzamide 0.22 g. MS m / z (ESI): 684.3 [M+H] + .

[0369] Step 4 Synthesis of 3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-N,N-dimethyl-5-(pentafluorothio)benzamide

[0370] 3-(2-(4-(4-ethoxy-6-((4-methoxybenzyl)oxy)pyridin-3-yl)-2-fluorophenyl)acetamido)-N,N-dimethyl-5-(pentafluorothio)benzamide (0.22 g, 0.32 mmol) was dissolved in DCM (6 mL), and then TFA (1 mL) was added, and the mixture was stirred at room temperature for 2 hours. After the reaction was complete, the reaction solution was concentrated, and then adjusted to neutral with sodium carbonate aqueous solution, extracted with DCM (10 mL*2), and the organic phase was dried by spin drying. Column chromatography gave 3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-N,N-dimethyl-5-(pentafluorothio)benzamide (Compound 2) 0.11 g. MS m / z(ESI): 564.5[M+H] + . 1 H NMR (500MHz, DMSO) δ11.39(s,1H),10.76(s,1H),8.29(t,J=2.0Hz,1H),7.85(s,1H),7.59(dd,J=1.9,1.3Hz,1H),7.40–7.32 (m,2H),7.28–7.20(m,2H),5.81(s,1H),4.04(q,J=7.0Hz,2H),3.78(s,2H),2.99(s,3H),2.90(s,3H),1.28(t,J=7.0Hz,3H).

[0371] Example 3 Synthesis of N-(2-(diethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(pentafluorothio)benzamide

[0372]

[0373] Step 1 Synthesis of N-(2-(diethylamino)ethyl)-3-nitro-5-(pentafluorothio)benzamide

[0374] Dissolve 3-nitro-5-(pentafluorothio)benzoic acid (0.3 g, 1.02 mmol) in SOCl2 (10 mL), add a few drops of DMF, and then reflux for 2 hours. After the reaction is complete, spin dry the reaction solution, dissolve the acid chloride in DCM (10 mL), cool the mixture to 0°C, then add triethylamine (0.32 g, 3.07 mmol), then add N,N-diethylethylenediamine (0.18 g, 1.56 mmol), and continue to react at 0°C for half an hour. After the reaction is complete, add water to quench the reaction, separate the liquids, concentrate the organic phase, and column chromatograph to obtain 0.32 g of N-(2-(diethylamino)ethyl)-3-nitro-5-(pentafluorothio)benzamide. MS m / z (ESI): 392.1 [M+H] + .

[0375] Step 2 Synthesis of 3-amino-N-(2-(diethylamino)ethyl)-5-(pentafluorothio)benzamide

[0376] N-(2-(diethylamino)ethyl)-3-nitro-5-(pentafluorothio)benzamide (0.32 g, 0.82 mmol) was added to methanol (10 mL), and then iron powder (0.8 g) was added, and then hydrochloric acid (1 mL, 12 mol / L) was slowly added, and the mixture was stirred at 72°C for 2 hours. After monitoring the reaction completion, it was cooled, filtered, and the reaction solution was spin-dried. Column chromatography was used to obtain 3-amino-N-(2-(diethylamino)ethyl)-5-((pentafluorothio)benzamide 0.25 g, MS m / z (ESI): 362.2 [M+H] + .

[0377] Step 3 Synthesis of N-(2-(diethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-((4-methoxybenzyl)oxy)pyridin-3-yl)-2-fluorophenyl)acetamido)-5-(pentafluorothio)benzamide

[0378] 2-(4(4-ethoxy-6-((4-methoxybenzyl)oxy)pyridin-3-yl)-2-fluorophenyl)acetic acid (0.25 g, 0.61 mmol) was added to pyridine (5 mL), and then 3-amino-N-(2-(diethylamino)ethyl)-5-(pentafluorothio)benzamide (0.26 g, 0.73 mmol) and T3P (0.28 g, 0.72 mmol) were added, and the mixture was reacted at room temperature for 12 hours. After the reaction was complete, the reaction solution was directly spin-dried and then column chromatographed to obtain 0.19 g of N-(2-(diethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-((4-methoxybenzyl)oxy)pyridin-3-yl)-2-fluorophenyl)acetamido)-5-(pentafluorothio)benzamide, MS m / z (ESI): 755.6 [M+H] + .

[0379] Step 4 Synthesis of N-(2-(diethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(pentafluorothio)benzamide

[0380] N-(2-(diethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-((4-methoxybenzyl)oxy)pyridin-3-yl)-2-fluorophenyl)acetamido)-5-(pentafluorothio)benzamide (0.19 g, 0.25 mmol) was dissolved in DCM (5 mL), and then TFA (1 mL) was added. The mixture was stirred at room temperature for 2 hours. After the reaction was complete, the reaction solution was concentrated and adjusted to neutral with aqueous sodium carbonate solution. DCM (10 mL*2) was used for extraction. The organic phase was dried by spin drying and column chromatography was performed to obtain 0.081 g of N-(2-(diethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(pentafluorothio)benzamide (Compound 3). MS m / z(ESI):635.1[M+H] + . 1 H NMR (400MHz, DMSO) δ11.58(s,1H),10.75(s,1H),8.67(s,1H),8.45(s,1H),8.24(s,1H),8.05(s,1H),7.40–7.32(m,2H),7.2 8–7.11(m,2H),5.83(s,1H),4.02(dt,J=5.8,2.1Hz,2H),3.78(s,4H),2.43(t,J=6.5Hz,4H),2.19(s,2H),1.17-1.35(t,9H).

[0381] Example 4 Synthesis of N-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetylamino)-N-methyl-5-(pentafluorothio)benzamide

[0382]

[0383] Step 1 Synthesis of N-(2-(dimethylamino)ethyl)-N-methyl-3-nitro-5-(pentafluorothio)benzamide

[0384] 3-Nitro-5-(pentafluorothio)benzoic acid (500 mg, 1.71 mmol) was dissolved in SOCl2 (5 mL), and a few drops of DMF were added. The mixture was refluxed for 2 hours. After the reaction was complete, the reaction solution was dried by spin drying. The acid chloride was dissolved in DCM (5 mL), and the mixture was added dropwise to triethylamine (517 mg, 5.12 mmol) and N 1 ,N 1 ,N 2 -trimethylethane-1,2-diamine (226 mg, 2.22 mmol) in DCM (5 mL) solution, continue to react at 0 ° C for half an hour. After the reaction is complete, water is added to quench the reaction, the liquid is separated, the organic phase is concentrated, and column chromatography is performed to obtain 460 mg of N-(2-(dimethylamino)ethyl)-N-methyl-3-nitro-5-(pentafluorothio)benzamide. MS m / z (ESI): 378 [M+H] + .

[0385] Step 2 Synthesis of 3-amino-N-(2-(dimethylamino)ethyl)-N-methyl-5-(pentafluorothio)benzamide

[0386] N-(2-(dimethylamino)ethyl)-N-methyl-3-nitro-5-(pentafluorothio)benzamide (460mg, 1.22mmol), FeCl3 (20mg, 0.12mmol), activated carbon (100mg) were added to ethanol (10mL), and then hydrazine hydrate (244mg, 4.88mmol) was slowly added to the reaction system, and the mixture was stirred at 80°C for overnight reaction. After monitoring the reaction was complete, it was cooled, filtered, and the reaction solution was spin-dried. Column chromatography was used to obtain 400mg of 3-amino-N-(2-(dimethylamino)ethyl)-N-methyl-5-(pentafluorothio)benzamide. MS m / z(ESI): 348[M+H] + .

[0387] Step 3 Synthesis of N-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-((4-methoxybenzyl)oxy)pyridin-3-yl)-2-fluorophenyl)acetamido)-N-methyl-5-(pentafluorothio)benzamide

[0388] 2-(4(4-ethoxy-6-((4-methoxybenzyl)oxy)pyridin-3-yl)-2-fluorophenyl)acetic acid (474 ​​mg, 1.15 mmol) was dissolved in DMF, and then HATU (482 mg, 1.27 mmol), DMAP (14Mg, 0.115 mmol), DIEA (446 mg, 3.46 mmol) and 3-amino-N-(2-(dimethylamino)ethyl)-N-methyl-5-(pentafluorothio)benzamide (400 mg, 1.15 mmol) were added, and the mixture was reacted at room temperature overnight. After the reaction was complete, water was added to quench the reaction, and then EA (20 mL*2) was used for extraction. The organic phase was dried, spin-dried, and column chromatography was performed to obtain 380 mg of N-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-((4-methoxybenzyl)oxy)pyridin-3-yl)-2-fluorophenyl)acetamido)-N-methyl-5-(pentafluorothio)benzamide. MS m / z (ESI): 741 [M+H] + .

[0389] Step 4 Synthesis of N-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetylamino)-N-methyl-5-(pentafluorothio)benzamide

[0390] N-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-((4-methoxybenzyl)oxy)pyridin-3-yl)-2-fluorophenyl)acetamido)-N-methyl-5-(pentafluorothio)benzamide (380 mg, 0.514 mmol) was dissolved in DCM (10 mL), and then TFA (2 mL) was added, and the mixture was stirred at room temperature for 2 hours. After the reaction was complete, the reaction solution was concentrated, and then adjusted to neutral with sodium carbonate aqueous solution, extracted with DCM (20 mL*2), and the organic phase was dried by spin drying. Column chromatography was performed to obtain 180 mg of N-(2-(dimethylamino)ethyl)-3-(2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-N-methyl-5-(pentafluorothio)benzamide (Compound 4). MS m / z(ESI):621[M+H] + . 1H NMR(400MHz,DMSO-d6)δ11.39(s,1H),10.79(s,1H),8.29(s,1H),7.85(s,1 H),7.56(s,1H),7.41–7.32(m,2H),7.29–7.21(m,2H),5.81(s,1H),4.05(q, J=6.9Hz,2H),3.79(s,2H),3.58(s,1H),3.24(s,1H),2.94(d,J=22.5Hz,3H) ,2.59(s,1H),2.37(s,1H),2.30(s,3H),1.95(s,3H),1.29(t,J=6.9Hz,3H).

[0391] Example 5 Synthesis of 2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)-N-(3-(pentafluorothio)-5-(pyrrolidine-1-carbonyl)phenyl)acetamide

[0392]

[0393] Step 1 Synthesis of (3-nitro-5-(pentafluorothio)phenyl)(pyrrolidin-1-yl)methanone

[0394] Dissolve 3-nitro-5-(pentafluorothio)benzoic acid (500mg, 1.71mmol) in dichlorothionyl (15mL), drip two drops of DMF into the system, heat to 80℃ and reflux for 2h. After the reaction is complete, spin dry the system and dissolve the acid chloride product with 1mL DCM. Pyrrolidine (243mg, 3.41mmol) and triethylamine (517mg, 5.12mmol) are dissolved in dichloromethane, and the acid chloride product is slowly dripped into the above system under ice bath. After completion, remove the ice bath and the reaction solution is reacted at room temperature for 30min. Wash with water, extract with dichloromethane, combine the organic phases, dry the organic phases, spin dry, and the crude product is chromatographed on a silica gel column with EA / PE=1:3 to obtain 480mg of the product (3-nitro-5-(pentafluorothio)phenyl)(pyrrolidin-1-yl)methanone. MS m / z (ESI): 347.3 [M+H] + .

[0395] Step 2 Synthesis of (3-amino-5-(pentafluorothio)phenyl)(pyrrolidin-1-yl)methanone

[0396] Under nitrogen protection, add (3-nitro-5-(pentafluorothio)phenyl)(pyrrolidin-1-yl)methanone (480mg, 1.39mmol), anhydrous ferric chloride (22.5mg, 0.14mmol), activated carbon (96mg, 6.0mmol) to a double-necked flask, add dry ethanol (20mL), reflux at 80°C, then slowly add hydrazine hydrate (278mg, 5.54mmol) dropwise, and continue to react for 2h. After monitoring the reaction is complete, filter the system and spin dry. The crude product is subjected to silica gel column chromatography with EA / PE=1:1 to obtain 376mg of pure product (3-amino-5-(pentafluorothio)phenyl)(pyrrolidin-1-yl)methanone. MS m / z (ESI): 333.3 [M+H] + .

[0397] Step 3 Synthesis of 2-(4-(4-ethoxy-6-(((4-methoxybenzyl)oxy)pyridin-3-yl)-2-fluorophenyl)-N-(3-(pentafluorothio)-5-(pyrrolidine-1-carbonyl)phenyl)acetamide

[0398] 2-(4-(4-ethoxy-6-((4-methoxybenzyl)oxy)pyridin-3-yl)-2-fluorophenyl)acetic acid (370 mg, 0.90 mmol) was dissolved in dry DMF (10 mL), HATU (376 mg, 0.99 mmol), (3-amino-5-(pentafluorothio)phenyl)(pyrrolidin-1-yl)methanone (300 mg, 0.90 mmol), DMAP (11 mg, 0.09 mmol) were added, and finally DIPEA (465 mg, 3.60 mmol) was added and stirred at room temperature for 2 h. After monitoring the reaction completion, the system was poured into ice water, extracted with EA, and the organic phases were combined and spin-dried. The crude product was purified by silica gel column chromatography using EA / PE=1:1 to obtain 370 mg of pure product 2-(4-(4-ethoxy-6-(((4-methoxybenzyl)oxy)pyridin-3-yl)-2-fluorophenyl)-N-(3-(pentafluorothio)-5-(pyrrolidine-1-carbonyl)phenyl)acetamide. MS m / z (ESI): 710.7 [M+H] + .

[0399] Step 4 Synthesis of 2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)-N-(3-(pentafluorothio)-5-(pyrrolidine-1-carbonyl)phenyl)acetamide

[0400] 2-(4-(4-ethoxy-6-(((4-methoxybenzyl)oxy)pyridin-3-yl)-2-fluorophenyl)-N-(3-(pentafluorothio)-5-(pyrrolidine-1-carbonyl)phenyl)acetamide (370 mg, 0.52 mmol) was dissolved in DCM, TFA (2 ml) was added, and the reaction was carried out at room temperature for 30 min. The reaction progress was monitored by TLC. After the reaction was completed, the system was spin-dried, water was added, and the organic phases were combined by dichloromethane extraction, dried and spin-dried. The crude product was subjected to silica gel column chromatography with EA / PE=2:1 to obtain the pure product 2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)-N-(3-(pentafluorothio)-5-(pyrrolidine-1-yl)carbonyl)phenyl)acetamide (Compound 5) 153.6 mg. MS m / z (ESI): 590.6 [M+H] + . 1 H NMR (500MHz, DMSO) δ11.36 (s, 1H), 10.75 (s, 1H), 8.28 (t, J = 2.0Hz, 1H), 7.98 (s, 1H), 7.66(s,1H),7.37(d,J=8.2Hz,1H),7.33(d,J=4.3Hz,1H),7.25(d,J=11.4Hz,1H),7. 22(dd,J=7.9,1.7Hz,1H),5.80(s,1H),4.03(q,J=7.0Hz,2H),3.78(s,1H),3.46(t,J =6.6Hz,2H),3.36(t,J=6.3Hz,2H),1.84(dt,J=18.4,6.5Hz,3H),1.35-1.14(m,4H).

[0401] Example 6 Synthesis of 2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)-N-(3-(pentafluorothio)-5-(piperidin-1-yl)carbonyl)phenyl)acetamide

[0402]

[0403] Step 1: Synthesis of (3-nitro-5-(pentafluorothio)phenyl)(piperidin-1-yl)methanone

[0404] Dissolve 3-nitro-5-(pentafluorothio)benzoic acid (500mg, 1.71mmol) in dichlorothionyl (15mL), then drop two drops of DMF into the system, heat to 80℃, and reflux for 2h. After the reaction is complete, spin dry the liquid in the system, and dissolve the acyl chloride product with 1mL DCM. Dissolve piperidine (290mg, 3.41mmol) and triethylamine (517mg, 5.12mmol) in dichloromethane, slowly drop the acyl chloride product into the above system under ice bath, remove the ice bath after completion, and react at room temperature for 30min. Wash with water, extract with dichloromethane, combine the organic phases, dry, and concentrate under reduced pressure. The crude product is chromatographed on a silica gel column with EA / PE=1:3 to obtain 350mg of the product (3-nitro-5-(pentafluorothio)phenyl)(piperidin-1-yl)methanone. MS m / z (ESI): 361.3 [M+H] + .

[0405] Step 2: Synthesis of (3-amino-5-(pentafluorothio)phenyl)(piperidin-1-yl)methanone

[0406] Under nitrogen protection, add (3-nitro-5-(pentafluorothio)phenyl)(piperidin-1-yl)methanone (485mg, 1.35mmol), ferrous chloride (21.83mg, 0.13mmol), activated carbon (97mg, 6.06mmol) to a double-necked flask, add dry ethanol (20mL), reflux at 80°C, then slowly add hydrazine hydrate (270.3mg, 5.40mmol), and continue to react for 2h. After monitoring the reaction is complete, filter the system and spin dry. The crude product is subjected to silica gel column chromatography with EA / PE=1:1 to obtain 404mg of pure product (3-amino-5-(pentafluorothio)phenyl)(piperidin-1-yl)methanone. MS m / z (ESI): 347.3 [M+H] + .

[0407] Step 3: Synthesis of 2-(4-(4-ethoxy-6-(((4-methoxybenzyl)oxy)pyridin-3-yl)-2-fluorophenyl)-N-(3-(pentafluorothio)-5-(piperidine-1-carbonyl)phenyl)acetamide

[0408] 2-(4-(4-ethoxy-6-((4-methoxybenzyl)oxy)pyridin-3-yl)-2-fluorophenyl)acetic acid (300 mg, 0.73 mmol) was dissolved in dry DMF (10 mL), HATU (305 mg, 0.8 mmol), (3-amino-5-(pentafluorothio)phenyl)(piperidin-1-yl)methanone (241 mg, 0.73 mmol), DMAP (9 mg, 0.07 mmol) were added, and finally DIPEA (377 mg, 2.92 mmol) was added and stirred at room temperature for 2 h. After monitoring the reaction completion, the system was poured into ice water, extracted with EA, and the organic phases were combined and spin-dried. The crude product was purified by silica gel column chromatography using EA / PE=1:1 to obtain 340 mg of pure product 2-(4-(4-ethoxy-6-(((4-methoxybenzyl)oxy)pyridin-3-yl)-2-fluorophenyl)-N-(3-(pentafluorothio)-5-(piperidine-1-carbonyl)phenyl)acetamide. MS m / z (ESI): 724.7 [M+H] + .

[0409] Step 4: Synthesis of 2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)-N-(3-(pentafluorothio)-5-(piperidin-1-yl)carbonyl)phenyl)acetamide

[0410] 2-(4-(4-ethoxy-6-(((4-methoxybenzyl)oxy)pyridin-3-yl)-2-fluorophenyl)-N-(3-(pentafluorothio)-5-(piperidin-1-carbonyl)phenyl)acetamide (113 mg, 0.16 mmol) was dissolved in DCM, TFA (2 ml) was added, and the reaction was carried out at room temperature for 30 min. The reaction progress was monitored by TLC. After the reaction was completed, the system was spin-dried, water was added, and the organic phases were combined by dichloromethane extraction. The organic phases were dried and spin-dried. The crude product was subjected to silica gel column chromatography using EA / PE=2:1 to obtain the pure product 2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)-N-(3-(pentafluorothio)-5-(piperidin-1-yl)carbonyl)phenyl)acetamide (Compound 6) 53.3 mg. MS m / z (ESI): 604.6 [M+H] + . 1H NMR (500MHz, DMSO) δ11.36 (s, 1H), 10.76 (s, 1H), 8.27 (t, J = 1.9Hz, 1H), 7. 83(s,1H),7.54(s,1H),7.37(d,J=8.1Hz,1H),7.33(d,J=3.7Hz,1H),7.25( d,J=11.4Hz,1H),7.30-7.20(m,1H),5.80(s,1H),4.03(q,J=6.9Hz,2H),3. 78(s,2H),3.57(s,2H),3.24(s,2H),1.60-1.46(m,4H),1.29-1.22(m,6H).

[0411] Example 7 Synthesis of N-(3-(3-(dimethylamino)azetidine-1-carbonyl)-5-(pentafluorothio)phenyl)-2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamide

[0412]

[0413] Step 1 Synthesis of (3-(dimethylamino)azetidin-1-yl)(3-nitro-5-(pentafluorothio)phenyl)methanone

[0414] 3-Nitro-5-(pentafluorothio)benzoic acid (500 mg, 1.71 mmol) was dissolved in SOCl2 (5 mL), and a few drops of DMF were added. The mixture was refluxed for 2 hours. After the reaction was complete, the reaction solution was dried and the acid chloride was dissolved in DCM (5 mL). The mixture was added dropwise to a DCM (5 mL) solution of triethylamine (517 mg, 5.12 mmol) and N,N-dimethylazetidine-3-amine (222 mg, 2.22 mmol), and the reaction was continued at 0°C for half an hour. After the reaction was complete, water was added to quench the reaction, the liquid was separated, the organic phase was concentrated, and column chromatography was performed to obtain 460 mg of (3-(dimethylamino)azetidine-1-yl)(3-nitro-5-(pentafluorothio)phenyl)methanone. MS m / z (ESI): 376 [M+H] + .

[0415] Step 2 Synthesis of (3-amino-5-(pentafluorothio)phenyl)(3-(dimethylamino)azetidin-1-yl)methanone

[0416] (3-(dimethylamino)azetidine-1-yl)(3-nitro-5-(pentafluorothio)phenyl)methanone (460 mg, 1.23 mmol), FeCl3 (20 mg, 0.12 mmol), and activated carbon (100 mg) were added to ethanol (10 mL), and then hydrazine hydrate (244 mg, 4.88 mmol) was slowly added, and the mixture was stirred at 80 ° C for overnight reaction. After monitoring the reaction was complete, it was cooled, filtered, and the reaction solution was spin-dried. Column chromatography gave 330 mg of (3-amino-5-(pentafluorothio)phenyl)(3-(dimethylamino)azetidine-1-yl)methanone. MS m / z (ESI): 346 [M+H] + .

[0417] Step 3 Synthesis of N-(3-(3-(dimethylamino)azetidine-1-carbonyl)-5-(pentafluorothio)phenyl)-2-(4-(4-ethoxy-6-((4-methoxybenzyl)oxy)pyridin-3-yl)-2-fluorophenyl)acetamide

[0418] 2-(4(4-ethoxy-6-((4-methoxybenzyl)oxy)pyridin-3-yl)-2-fluorophenyl)acetic acid (393 mg, 0.957 mmol) was dissolved in DMF, and HATU (400 mg, 1.05 mmol), DMAP (12 mg, 0.095 mmol), DIEA (370 mg, 2.87 mmol), (3-amino-5-(pentafluorothio)phenyl)(3-(dimethylamino)azetidine-1-yl)methanone (330 mg, 0.957 mmol) were added, and the mixture was reacted at room temperature overnight. After the reaction was complete, water was added to quench the reaction, and then EA (20 mL*2) was used for extraction. The organic phase was dried, spin-dried, and column chromatography was performed to obtain 300 mg of N-(3-(3-(dimethylamino)azetidine-1-carbonyl)-5-(pentafluorothio)phenyl)-2-(4-(4-ethoxy-6-((4-methoxybenzyl)oxy)pyridin-3-yl)-2-fluorophenyl)acetamide. MS m / z (ESI): 739 [M+H] + .

[0419] Step 4 Synthesis of N-(3-(3-(dimethylamino)azetidine-1-carbonyl)-5-(pentafluorothio)phenyl)-2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamide

[0420] N-(3-(3-(dimethylamino)azetidine-1-carbonyl)-5-(pentafluorothio)phenyl)-2-(4-(4-ethoxy-6-((4-methoxybenzyl)oxy)pyridin-3-yl)-2-fluorophenyl)acetamide (300 mg, 0.407 mmol) was dissolved in DCM (10 mL), and then TFA (2 mL) was added, and the mixture was stirred at room temperature for 2 hours. After the reaction was complete, the reaction solution was concentrated, and then adjusted to neutral with sodium carbonate aqueous solution, extracted with DCM (20 mL*2), and the organic phase was dried by spin drying. Column chromatography gave 85 mg of N-(3-(3-(dimethylamino)azetidine-1-carbonyl)-5-(pentafluorothio)phenyl)-2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamide (Compound 7). MS m / z(ESI):619[M+H] + . 1 H NMR (500MHz, DMSO-d6) δ11.40(s,1H),10.80(s,1H),8.40(t,J=2.0Hz,1H),8.05( t,J=1.5Hz,1H),7.71(t,J=1.6Hz,1H),7.40–7.32(m,2H),7.28–7.20(m,2H),5.81 (s,1H),4.30(t,J=8.0Hz,1H),4.05(dq,J=14.0,7.5,7.0Hz,4H),3.85(dd,J=10. 5,5.1Hz,1H),3.79(s,2H),3.14–3.07(m,1H),2.08(s,6H),1.28(t,J=6.9Hz,3H).

[0421] Example 8 Synthesis of 2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)-N-(3-(morpholine-4-carbonyl)-5-(pentafluorothio)phenyl)acetamide

[0422]

[0423] Step 1 Synthesis of morpholino (3-nitro-5-(pentafluorothio)phenyl)methanone

[0424] Dissolve 3-nitro-5-(pentafluorothio)benzoic acid (500mg, 1.71mmol) in dichlorothionyl (15mL), drip two drops of DMF into the system, heat to 80℃ and reflux for 2h. After the reaction is complete, spin dry the system and dissolve the acyl chloride product with 1mL DCM. Morpholine (298mg, 3.41mmol) and triethylamine (517mg, 5.12mmol) are dissolved in dichloromethane, and the acyl chloride product is slowly dripped into the above system under ice bath. After completion, remove the ice bath and react at room temperature for 30min. Wash with water, extract with dichloromethane, combine the organic phases, dry and spin dry, and the crude product is chromatographed on a silica gel column with EA / PE=1:3 to obtain 354mg of the product morpholino (3-nitro-5-(pentafluorothio)phenyl)methanone. MS m / z (ESI): 363.3 [M+H] + .

[0425] Step 2 Synthesis of morpholino (3-amino-5- (pentafluorothio) phenyl) ketone

[0426] Under nitrogen protection, add morpholino (3-amino-5-(pentafluorothio)phenyl)methanone (354mg, 0.98mmol), anhydrous ferric chloride (15.9mg, 0.10mmol), activated carbon (70.8mg, 4.42mmol), and dry ethanol (20mL) to a double-necked flask, reflux at 80°C, then slowly add hydrazine hydrate (196mg, 3.91mmol) dropwise, and continue to react for 2h. After monitoring the reaction is complete, filter the system and spin dry. The crude product is subjected to silica gel column chromatography with EA / PE=1:1 to obtain 307mg of pure product morpholino (3-amino-5-(pentafluorothio)phenyl)methanone. MS m / z (ESI): 349.3 [M+H] + .

[0427] Step 3 Synthesis of 2-(4-(4-ethoxy-6-(((4-methoxybenzyl)oxy)pyridin-3-yl)-2-fluorophenyl)-N-(3-(morpholine-4-carbonyl)-5-(pentafluorothio)phenyl)acetamide

[0428] 2-(4-(4-ethoxy-6-((4-methoxybenzyl)oxy)pyridin-3-yl)-2-fluorophenyl)acetic acid (362 mg, 0.88 mmol) was dissolved in dry DMF (10 mL), HATU (368 mg, 0.97 mmol), morpholino(3-amino-5-(pentafluorothio)phenyl)methanone (307 mg, 0.88 mmol), DMAP (10.8 mg, 0.09 mmol) were added, and finally DIPEA (455 mg, 3.52 mmol) was added and stirred at room temperature for 2 h. After monitoring the reaction completion, the system was poured into ice water, extracted with EA, and the organic phases were combined and spin-dried. The crude product was subjected to silica gel column chromatography using EA / PE=1:1 to obtain 600 mg of the pure product 2-(4-(4-ethoxy-6-(((4-methoxybenzyl)oxy)pyridin-3-yl)-2-fluorophenyl)-N-(3-(morpholine-4-carbonyl)-5-(pentafluorothio)phenyl)acetamide. MS m / z (ESI): 726.7 [M+H] + .

[0429] Step 4 Synthesis of 2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)-N-(3-(morpholine-4-carbonyl)-5-(pentafluorothio)phenyl)acetamide

[0430] 2-(4-(4-ethoxy-6-(((4-methoxybenzyl)oxy)pyridin-3-yl)-2-fluorophenyl)-N-(3-(morpholine-4-carbonyl)-5-(pentafluorothio)phenyl)acetamide (600 mg, 0.83 mmol) was dissolved in DCM, TFA (2 mL) was added, and the reaction was carried out at room temperature for 30 min. The reaction progress was monitored by TLC. After the reaction was completed, the system was spin-dried, water was added, and the organic phases were combined by dichloromethane extraction and dried. The crude product was subjected to silica gel column chromatography using EA / PE=2:1 to obtain the pure product 2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)-N-(3-(pentafluorothio)-5-(pyrrolidin-1-yl)carbonyl)phenyl)acetamide (Compound 8) 225 mg. MS m / z (ESI): 606.6 [M+H]+. 1H NMR (500MHz, DMSO) δ11.37(s,1H),10.77(s,1H),8.29(d,J=1.9Hz,1H),7.84(s,1H),7.61(s,1H),7.37(d,J=8.1Hz,1H),7.33(d,J=3. 6Hz,1H),7.25(d,J=11.5Hz,1H),7.20(m,1H),5.80(s,1H),4.03(q,J=6.9Hz,2H),3.78(s,1H),3.62-3.56(m,4H),1.37-1.18(m,3H).

[0431] Example 9 Synthesis of N-(3-(3-(dimethylamino)pyrrolidine-1-carbonyl)-5-(pentafluorothio)phenyl)-2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamide

[0432]

[0433] Step 1 Synthesis of (3-(dimethylamino)pyrrolidin-1-yl)(3-nitro-5-(pentafluorothio)phenyl)methanone

[0434] 3-Nitro-5-(pentafluorothio)benzoic acid (500 mg, 1.71 mmol) was dissolved in SOCl2 (5 mL), and a few drops of DMF were added. The mixture was refluxed for 2 hours. After the reaction was complete, the reaction solution was dried by spin drying. The acid chloride was dissolved in DCM (5 mL), and the mixture was added dropwise to a DCM (5 mL) solution of triethylamine (517 mg, 5.12 mmol) and N,N-dimethylpyrrolidinobutyl-3-amine (253 mg, 2.22 mmol), and the reaction was carried out at 0°C for half an hour. After the reaction was complete, water was added to quench the reaction, the liquid was separated, the organic phase was concentrated, and column chromatography was performed to obtain 480 mg of ((3-(dimethylamino)pyrrolidin-1-yl)(3-nitro-5-(pentafluorothio)phenyl)methanone. MS m / z (ESI): 390 [M+H] + .

[0435] Step 2 Synthesis of (3-amino-5-(pentafluorothio)phenyl)(3-(dimethylamino)pyrrolidin-1-yl)methanone

[0436] (3-(dimethylamino)pyrrolidin-1-yl)(3-nitro-5-(pentafluorothio)phenyl)methanone (480 mg, 1.23 mmol), FeCl3 (20 mg, 0.12 mmol), and activated carbon (100 mg) were added to ethanol (10 mL), and hydrazine hydrate (247 mg, 4.93 mmol) was slowly added to the system. The mixture was stirred and reacted at 80°C overnight. After monitoring the reaction was complete, it was cooled, filtered, and the reaction solution was spin-dried. Column chromatography was used to obtain 375 mg of (3-amino-5-(pentafluorothio)phenyl)(3-(dimethylamino)pyrrolidin-1-yl)methanone. MS m / z (ESI): 360 [M+H] + .

[0437] Step 3 Synthesis of N-(3-(3-(dimethylamino)pyrrolidine-1-carbonyl)-5-(pentafluorothio)phenyl)-2-(4-(4-ethoxy-6-((4-methoxybenzyl)oxy)pyridin-3-yl)-2-fluorophenyl)acetamide

[0438] 2-(4(4-ethoxy-6-((4-methoxybenzyl)oxy)pyridin-3-yl)-2-fluorophenyl)acetic acid (429 mg, 1.045 mmol) was dissolved in DMF, and then HATU (437 mg, 1.15 mmol), DMAP (13 mg, 0.104 mmol), DIEA (404 mg, 3.13 mmol), (3-amino-5-(pentafluorothio)phenyl)(3-(dimethylamino)pyrrolidin-1-yl)methanone (375 mg, 1.045 mmol) were added, and the mixture was reacted at room temperature overnight. After the reaction was complete. Water was added to quench the reaction, and then EA (20 mL*2) was used for extraction. The organic phase was dried, spin-dried, and column chromatography was performed to obtain 320 mg of N-(3-(3-(dimethylamino)pyrrolidine-1-carbonyl)-5-(pentafluorothio)phenyl)-2-(4-(4-ethoxy-6-((4-methoxybenzyl)oxy)pyridin-3-yl)-2-fluorophenyl)acetamide. MS m / z (ESI): 753 [M+H] + .

[0439] Step 4 Synthesis of N-(3-(3-(dimethylamino)pyrrolidine-1-carbonyl)-5-(pentafluorothio)phenyl)-2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamide

[0440] N-(3-(3-(dimethylamino)pyrrolidine-1-carbonyl)-5-(pentafluorothio)phenyl)-2-(4-(4-ethoxy-6-((4-methoxybenzyl)oxy)pyridin-3-yl)-2-fluorophenyl)acetamide (320 mg, 0.426 mmol) was dissolved in DCM (10 mL), and then TFA (2 mL) was added, and the mixture was stirred at room temperature for 2 hours. After the reaction was complete, the reaction solution was concentrated, and then adjusted to neutral with sodium carbonate aqueous solution, extracted with DCM (20 mL*2), and the organic phase was dried by spin drying. Column chromatography was performed to obtain 200 mg of N-(3-(3-(dimethylamino)pyrrolidine-1-carbonyl)-5-(pentafluorothio)phenyl)-2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamide (Compound 9). MS m / z(ESI):633[M+H] + . 1 H NMR (400 MHz, DMSO-d 6 )δ11.40(s,1H),10.89(s,1H),8.31(q,J=3.0,1.9Hz,1H),8.03(d,J=14.1Hz,1H ),7.69(s,1H),7.36(d,J=10.5Hz,2H),7.29–7.21(m,2H),5.82(s,1H),4.05(q, J=7.0Hz,2H),3.90(dd,J=12.6,7.1Hz,1H),3.80(s,2H),3.53(q,J=17.8,14.1H z, 4H), 2.68 (d, J = 60.3Hz, 6H), 2.27 (s, 1H), 2.08 (s, 1H), 1.29 (t, J = 6.9Hz, 3H).

[0441] Example 10 Synthesis of N-(2-(dimethylamino)ethyl)-3-(2-(2-fluoro-4-(4-isobutoxy-6-oxo-1,6-dihydropyridin-3-yl)phenyl)acetamido)-5-(pentafluorothio)benzamide

[0442]

[0443] Step 1 Synthesis of 2-chloro-4-isobutoxypyridine

[0444] Dissolve isobutyl alcohol (4.68 g, 63.29 mmol) in THF (50 mL), add NaH (60%) (3.16 g, 79.11 mmol) under ice bath and stir for 30 minutes. Then add 2-chloro-4-nitropyridine (5 g, 31.65 mmol) and stir overnight. After the reaction is complete, spin dry the reaction solution, extract with ethyl acetate, dry, spin dry, and column chromatography to obtain 3.6 g of 2-chloro-4-isobutoxypyridine. MSm / z (ESI): 186 [M+H] + .

[0445] Step 2 Synthesis of 5-bromo-2-chloro-4-isobutoxypyridine

[0446] Dissolve 2-chloro-4-isobutoxypyridine (3.6 g, 19.46 mmol) in concentrated sulfuric acid (20 mL), then slowly add NBS (5.2 g, 29.19 mmol) to the system in batches. After the addition, heat the reaction solution to 80 °C and react for 4 hours. After the reaction is complete, cool the reaction solution and pour it into ice water. Extract with EA (50 mL*3), dry the organic phase, and perform column chromatography to obtain 1.1 g of 5-bromo-2-chloro-4-isobutoxypyridine. MS m / z (ESI): 264 [M+H] + .

[0447] Step 3 Synthesis of 5-bromo-4-isobutoxy-2-((4-methoxybenzyl)oxy)pyridine

[0448] At room temperature, (4-methoxyphenyl)methanol (693mg, 5.02mmol), KOH (469mg, 8.37mmol) and 18-crown-6 (110mg, 0.42mmol) were added to a mixture of 5-bromo-2-chloro-4-ethoxypyridine (1.1g, 4.18mmol) in toluene (20mL), and the mixture was reacted at 120°C for 3 hours. After the reaction was complete, the reaction solution was concentrated under reduced pressure, water (15mL) was added, and then extracted with EA (20mL*2), the organic phase was dried, spin-dried, and column chromatography was performed to obtain 950mg of 5-bromo-4-isobutoxy-2-((4-methoxybenzyl)oxy)pyridine. MS m / z (ESI): 366 [M+H] + .

[0449] Step 4 Synthesis of methyl 2-(2-fluoro-4-(4-isobutoxy-6-((4-methoxybenzyl)oxy)pyridin-3-yl)phenyl)acetate

[0450] Under nitrogen protection, 5-bromo-4-isobutoxy-2-((4-methoxybenzyl)oxy)pyridine (950 mg, 2.6 mmol), methyl 2-(2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)acetate (842 mg, 2.86 mmol), Pd(dppf)Cl2 (212 mg, 0.26 mmol) and cesium carbonate (1.7 g, 5.2 mmol) were added to a single-necked bottle, and 1,4-dioxane (15 mL) and water (5 mL) were added, and the mixture was reacted at 95 ° C for 3 hours. After the reaction was complete, EA (20 mL*2) was used for extraction, the organic phase was dried, concentrated under reduced pressure and dried by spin drying, and column chromatography was performed to obtain 980 mg of methyl 2-(2-fluoro-4-(4-isobutoxy-6-((4-methoxybenzyl)oxy)pyridin-3-yl)phenyl)acetate. MS m / z (ESI): 454 [M+H] + .

[0451] Step 5 Synthesis of 2-(2-fluoro-4-(4-isobutoxy-6-((4-methoxybenzyl)oxy)pyridin-3-yl)phenyl)acetic acid

[0452] 2-(2-fluoro-4-(4-isobutoxy-6-((4-methoxybenzyl)oxy)pyridin-3-yl)phenyl)acetic acid methyl ester (980mg, 2.16mmol) was dissolved in THF (15mL), and then lithium hydroxide (182mg, 4.33mmol) aqueous solution was added to the reaction system, and the mixture was stirred at 60°C for 4 hours. After the reaction was complete, most of the organic phase was removed by concentration under reduced pressure, and the pH of the system was adjusted to 7 by adding dilute hydrochloric acid. A white solid was precipitated, filtered, the filter cake was washed with water, and dried to obtain 670mg of 2-(2-fluoro-4-(4-isobutoxy-6-((4-methoxybenzyl)oxy)pyridin-3-yl)phenyl)acetic acid. MS m / z(ESI): 440[M+H] + .

[0453] Step 6 Synthesis of N-(2-(dimethylamino)ethyl)-3-(2-(2-fluoro-4-(4-isobutoxy-6-((4-methoxybenzyl)oxy)pyridin-3-yl)phenyl)acetylamino)-5-(pentafluorothio)benzamide

[0454] 2-(2-Fluoro-4-(4-isobutoxy-6-((4-methoxybenzyl)oxy)pyridin-3-yl)phenyl)acetic acid (670 mg, 1.53 mmol) was dissolved in DMF, and then HATU (638 mg, 1.68 mmol), DMAP (19 mg, 0.153 mmol), DIEA (636 mg, 4.58 mmol), 3-amino-N-(2-(dimethylamino)ethyl)-5-(pentafluorosulfuryl)- The mixture was reacted overnight at room temperature. After the reaction was complete, water was added to quench the reaction, and then extracted with EA (20 mL*2). The organic phase was dried, spin-dried, and column chromatography was performed to obtain 380 mg of N-(2-(dimethylamino)ethyl)-3-(2-(2-fluoro-4-(4-isobutoxy-6-((4-methoxybenzyl)oxy)pyridin-3-yl)phenyl)acetylamino)-5-(pentafluorothio)benzamide. MS m / z(ESI): 755[M+H] + .

[0455] Step 7 Synthesis of N-(2-(dimethylamino)ethyl)-3-(2-(2-fluoro-4-(4-isobutoxy-6-oxo-1,6-dihydropyridin-3-yl)phenyl)acetamido)-5-(pentafluorothio)benzamide

[0456] N-(2-(dimethylamino)ethyl)-3-(2-(2-fluoro-4-(4-isobutoxy-6-((4-methoxybenzyl)oxy)pyridin-3-yl)phenyl)acetamido)-5-(pentafluorothio)benzamide (380 mg, 0.504 mmol) was dissolved in DCM (10 mL), and then TFA (2 mL) was added, and the mixture was stirred at room temperature for 2 hours. After the reaction was complete, the reaction solution was concentrated, and then adjusted to neutral with sodium carbonate aqueous solution, extracted with DCM (20 mL*2), and the organic phase was removed by concentration under reduced pressure. Column chromatography gave 190 mg of N-(2-(dimethylamino)ethyl)-3-(2-(2-fluoro-4-(4-isobutoxy-6-oxo-1,6-dihydropyridin-3-yl)phenyl)acetamido)-5-(pentafluorothio)benzamide (Compound 10). MS m / z(ESI):635[M+H] + . 1 H NMR (400 MHz, DMSO-d 6)δ11.39(s,1H),10.86(s,1H),8.79(t,J=5.7Hz,1H),8.45(t,J=2.0Hz,1H ),8.26(d,J=1.6Hz,1H),8.03(t,J=1.7Hz,1H),7.42–7.34(m,2H),7.28–7 .21(m,2H),5.81(s,1H),3.80(s,2H),3.77(d,J=6.3Hz,2H),3.40(q,J=6. 4Hz, 4H), 2.24 (s, 6H), 1.97 (dt, J = 13.2, 6.6Hz, 1H), 0.92 (d, J = 6.7Hz, 6H).

[0457] Example 11 Synthesis of N-(2-(dimethylamino)ethyl)-3-(2-(4-(5-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(pentafluorothio)benzamide

[0458]

[0459] Step 1 Synthesis of compound 5-bromo-2-chloro-3-ethoxypyridine

[0460] 5-Bromo-2-chloropyridin-3-ol (1g, 4.83mmol), iodoethane (980mg, 6.28mmol), potassium iodide (80mg, 0.483mmol) and potassium carbonate (1.33g, 9.67mmol) were dissolved in DMF (20mL), and the mixture was reacted at 50°C overnight. After the reaction was complete, the reaction solution was poured into ice water, extracted with ethyl acetate, and the organic phase was dried, spin-dried, and column chromatography was performed to obtain 1.1g of 5-bromo-2-chloro-3-ethoxypyridine. MS m / z (ESI): 236 [M+H] + .

[0461] Step 2 Synthesis of 5-bromo-3-ethoxy-2-((4-methoxybenzyl)oxy)pyridine

[0462] 5-Bromo-2-chloro-3-ethoxypyridine (1.21 g, 5.12 mmol), (4-methoxyphenyl)methanol (848 mg, 6.14 mmol), 18-crown ether-6 (135 mg, 0.51 mmol), potassium hydroxide (574 mg, 5.23 mmol) were dissolved in toluene (15 mL) and heated to 120 ° C for 5 h. After the reaction was complete, water was added to the system to quench, EA was extracted, the organic phases were combined, the organic phases were dried, and the crude product was subjected to silica gel column chromatography with EA / PE = 1:3 to obtain 1.26 g of the product 5-bromo-3-ethoxy-2-((4-methoxybenzyl)oxy)pyridine. MS m / z (ESI): 339.2 [M+H] + .

[0463] Step 3 Synthesis of methyl 2-(4-(5-ethoxy-6-(((4-methoxybenzyl)oxy)pyridin-3-yl)-2-fluorophenyl)acetate

[0464] 5-Bromo-3-ethoxy-2-((4-methoxybenzyl)oxy)pyridine (1.26 g, 3.73 mmol), methyl 2-(2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)acetate (1.21 g, 4.10 mmol), Pd(dppf)Cl2 (270 mg, 0.37 mmol), and Cs2CO3 (2.43 g, 7.45 mmol) were dissolved in a mixed solution of 1,4-dioxane / H2O (40 mL, 3:1), and heated to 85°C and refluxed for reaction overnight. After the reaction was complete, water was added to quench the reaction, and the mixture was extracted with EA. The organic phases were combined, dried, and spun off. The crude product was chromatographed on a silica gel column using EA / PE=1:2 to obtain 1.34 g of methyl 2-(4-(5-ethoxy-6-(((4-methoxybenzyl)oxy)pyridin-3-yl)-2-fluorophenyl)acetate. MS m / z (ESI): 426.5 [M+H] + .

[0465] Step 4 Synthesis of 2-(4-(5-ethoxy-6-(((4-methoxybenzyl)oxy)pyridin-3-yl)-2-fluorophenyl)acetic acid

[0466] 2-(4-(5-ethoxy-6-(((4-methoxybenzyl)oxy)pyridin-3-yl)-2-fluorophenyl)acetic acid methyl ester (1.34 g, 3.15 mmol) and lithium hydroxide (260 mg, 6.30 mmol) were dissolved in MeOH / H2O (20 mL, 3:1) and reacted at room temperature for 2 h. After monitoring the completion of the reaction, the system was spin-dried, washed with water, adjusted to a weakly acidic pH with dilute hydrochloric acid, and extracted with EA. The organic phases were combined and spin-dried. The crude product was subjected to silica gel column chromatography using EA / PE=1:1 to obtain 1.25 g of pure product 2-(4-(5-ethoxy-6-(((4-methoxybenzyl)oxy)pyridin-3-yl)-2-fluorophenyl)acetic acid. MS m / z(ESI): 412.4[M+H] + .

[0467] Step 5: Synthesis of N-(2-(dimethylamino)ethyl)-3-(2-(4-(5-ethoxy-6-((4-methoxybenzyl)oxy)pyridin-3-yl)-2-fluorophenyl)acetamido)-5-(pentafluorothio)benzamide

[0468] 2-(4-(5-ethoxy-6-(((4-methoxybenzyl)oxy)pyridin-3-yl)-2-fluorophenyl)acetic acid (300 mg, 0.73 mmol) was dissolved in dry DMF (10 mL), HATU (305 mg, 0.8 mmol), 3-amino-N-(2-(dimethylamino)ethyl)-5-(pentafluorothio)benzamide (243 mg, 0.73 mmol), DMAP (9.0 mg, 0.07 mmol) were added, and finally D IPEA (377 mg, 2.92 mmol), stirred at room temperature for 2 h. After monitoring the reaction completion, the system was poured into ice water, extracted with EA, the organic phases were combined and dried. The crude product was subjected to silica gel column chromatography with DCM / MeOH=10:1 to obtain 237 mg of pure product N-(2-(dimethylamino)ethyl)-3-(2-(4-(5-ethoxy-6-((4-methoxybenzyl)oxy)pyridin-3-yl)-2-fluorophenyl)acetylamino)-5-(pentafluorothio)benzamide. MS m / z(ESI): 727.7[M+H] + .

[0469] Step 6: Synthesis of N-(2-(dimethylamino)ethyl)-3-(2-(4-(5-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(pentafluorothio)benzamide

[0470] N-(2-(dimethylamino)ethyl)-3-(2-(4-(5-ethoxy-6-((4-methoxybenzyl)oxy)pyridin-3-yl)-2-fluorophenyl)acetamido)-5-(pentafluorothio)benzamide (191 mg, 0.26 mmol) was dissolved in DCM, TFA (2 mL) was added, and the reaction was carried out at room temperature for 30 min. The reaction progress was monitored by TLC. After the reaction was complete, the system was spin-dried, water was added, and the organic phases were combined by dichloromethane extraction, dried and spin-dried. The crude product was subjected to silica gel column chromatography with DCM / MeOH=10:1 to obtain 22 mg of the pure product N-(2-(dimethylamino)ethyl)-3-(2-(4-(5-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamido)-5-(pentafluorothio)benzamide (Compound 11). MS m / z(ESI):607.6[M+H] + . 1 H NMR (500MHz, DMSO) δ11.84(s,1H),10.83(s,1H),8.81(s,1H),8.43(s,1H),8.25(s,1H),8.01(s,1H),7.47(d,J=12.0Hz,1H),7.40(d,J=5 .4Hz,1H),7.36(d,J=9.0Hz,1H),7.14(d,J=2.1Hz,1H),4.03(q,J=7.0Hz,2H),3.78(s,2H),2.63(m,2H),2.31(s,4H),1.48-1.15(m,4H).

[0471] Example 12 Synthesis of (S)-N-(3-(3-(dimethylamino)pyrrolidine-1-carbonyl)-5-(pentafluorothio)phenyl)-2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamide

[0472]

[0473] Step 1 Synthesis of (S)-(3-(dimethylamino)pyrrolidin-1-yl)(3-nitro-5-(pentafluorothio)phenyl)methanone

[0474] 3-Nitro-5-(pentafluorothio)benzoic acid (500 mg, 1.71 mmol) was dissolved in SOCl2 (5 mL), and a few drops of DMF were added. The mixture was refluxed for 2 hours. After the reaction was complete, the reaction solution was dried, the acid chloride was dissolved in DCM (5 mL), and the mixture was added dropwise to a DCM (5 mL) solution of triethylamine (517 mg, 5.12 mmol) and (S)-N,N-dimethylpyrrolidin-3-amine (253 mg, 2.22 mmol), and the reaction was continued at 0°C for half an hour. After the reaction was complete, water was added to quench the reaction, the liquid was separated, the organic phase was concentrated, and column chromatography was performed to obtain 470 mg of (S)-(3-(dimethylamino)pyrrolidin-1-yl)(3-nitro-5-(pentafluorothio)phenyl)methanone. MS m / z (ESI): 390 [M+H] + .

[0475] Step 2 Synthesis of (S)-(3-amino-5-(pentafluorothio)phenyl)(3-(dimethylamino)pyrrolidin-1-yl)methanone

[0476] (S)-(3-(dimethylamino)pyrrolidin-1-yl)(3-nitro-5-(pentafluorothio)phenyl)methanone (470 mg, 1.23 mmol), FeCl3 (20 mg, 0.12 mmol), and activated carbon (100 mg) were added to ethanol (10 mL), and hydrazine hydrate (242 mg, 4.83 mmol) was slowly added to the system. The mixture was stirred and reacted at 80°C overnight. After monitoring the reaction was complete, it was cooled, filtered, and the reaction solution was spin-dried. Column chromatography was performed to obtain 360 mg of (S)-(3-amino-5-(pentafluorothio)phenyl)(3-(dimethylamino)pyrrolidin-1-yl)methanone. MS m / z(ESI): 360[M+H] + .

[0477] Step 3 Synthesis of (S)-N-(3-(3-(dimethylamino)pyrrolidine-1-carbonyl)-5-(pentafluorothio)phenyl)-2-(4-(4-ethoxy-6-((4-methoxybenzyl)oxy)pyridin-3-yl)-2-fluorophenyl)acetamide

[0478] 2-(4(4-ethoxy-6-((4-methoxybenzyl)oxy)pyridin-3-yl)-2-fluorophenyl)acetic acid (412 mg, 1.003 mmol) was dissolved in DMF, and then HATU (419 mg, 1.103 mmol), DMAP (12 mg, 0.100 mmol), DIEA (388 mg, 3.008 mmol), (S)-(3-amino-5-(pentafluorothio)phenyl)(3-(dimethylamino)pyrrolidin-1-yl)methanone (360 mg, 1.003 mmol) were added, and the mixture was reacted at room temperature overnight. After the reaction was complete, water was added to quench the reaction, and then EA (20 mL*2) was used for extraction. The organic phase was dried, spin-dried, and column chromatography was performed to obtain 310 mg of (S)-N-(3-(3-(dimethylamino)pyrrolidine-1-carbonyl)-5-(pentafluorothio)phenyl)-2-(4-(4-ethoxy-6-((4-methoxybenzyl)oxy)pyridin-3-yl)-2-fluorophenyl)acetamide. MS m / z (ESI): 753 [M+H] + .

[0479] Step 4 Synthesis of (S)-N-(3-(3-(dimethylamino)pyrrolidine-1-carbonyl)-5-(pentafluorothio)phenyl)-2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamide

[0480] (S)-N-(3-(3-(dimethylamino)pyrrolidine-1-carbonyl)-5-(pentafluorothio)phenyl)-2-(4-(4-ethoxy-6-((4-methoxybenzyl)oxy)pyridin-3-yl)-2-fluorophenyl)acetamide (310 mg, 0.412 mmol) was dissolved in DCM (10 mL), and then TFA (2 mL) was added, and the mixture was stirred at room temperature for 2 hours. After the reaction was complete, the reaction solution was concentrated, and then adjusted to neutral with sodium carbonate aqueous solution, extracted with DCM (20 mL*2), and the organic phase was dried by spin drying. Column chromatography was performed to obtain 170 mg of (S)-N-(3-(3-(dimethylamino)pyrrolidine-1-carbonyl)-5-(pentafluorothio)phenyl)-2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamide (Compound 12). MS m / z(ESI):633[M+H] + . 1 H NMR (400 MHz, DMSO-d 6)δ11.40(s,1H),10.91(s,1H),8.36–8.30(m,1H),8.02(d,J=13.8Hz,1H),7.69 (d,J=2.6Hz,1H),7.37(d,J=10.5Hz,2H),7.29–7.21(m,2H),5.82(s,1H),4.05 (q,J=6.9Hz,2H),3.86(d,J=5.2Hz,1H),3.81(s,2H),3.56–3.46(m,4H),2.63( s,3H),2.46(s,3H),2.20(d,J=13.1Hz,1H),2.00(s,1H),1.29(t,J=7.0Hz,3H).

[0481] Example 13 Synthesis of (R)-N-(3-(3-(dimethylamino)pyrrolidine-1-carbonyl)-5-(pentafluorothio)phenyl)-2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamide

[0482]

[0483] Step 1 Synthesis of (R)-(3-(dimethylamino)pyrrolidin-1-yl)(3-nitro-5-(pentafluorothio)phenyl)methanone

[0484] 3-Nitro-5-(pentafluorothio)benzoic acid (500 mg, 1.71 mmol) was dissolved in SOCl2 (5 mL), and a few drops of DMF were added. The mixture was refluxed for 2 hours. After the reaction was complete, the reaction solution was dried, the acid chloride was dissolved in DCM (5 mL), and the mixture was added dropwise to a DCM (5 ml) solution of triethylamine (517 mg, 5.12 mmol) and (R)-N,N-dimethylpyrrolidin-3-amine (253 mg, 2.22 mmol), and the reaction was continued at 0°C for half an hour. After the reaction was complete, water was added to quench the reaction, the liquid was separated, the organic phase was concentrated, and column chromatography was performed to obtain 560 mg of (R)-(3-(dimethylamino)pyrrolidin-1-yl)(3-nitro-5-(pentafluorothio)phenyl)methanone. MS m / z (ESI): 390 [M+H] + .

[0485] Step 2 Synthesis of (R)-(3-amino-5-(pentafluorothio)phenyl)(3-(dimethylamino)pyrrolidin-1-yl)methanone

[0486] (R)-(3-(dimethylamino)pyrrolidin-1-yl)(3-nitro-5-(pentafluorothio)phenyl)methanone (560 mg, 1.23 mmol), FeCl3 (23 mg, 0.14 mmol), and activated carbon (100 mg) were added to ethanol (10 mL), and then hydrazine hydrate (288 mg, 5.76 mmol) was slowly added. The mixture was stirred at 80°C for overnight reaction. After monitoring the reaction completion, the mixture was cooled, filtered, and the reaction solution was spin-dried. Column chromatography was performed to obtain 420 mg of (R)-(3-amino-5-(pentafluorothio)phenyl)(3-(dimethylamino)pyrrolidin-1-yl)methanone. MS m / z(ESI): 360[M+H] + .

[0487] Step 3 Synthesis of (R)-N-(3-(3-(dimethylamino)pyrrolidine-1-carbonyl)-5-(pentafluorothio)phenyl)-2-(4-(4-ethoxy-6-((4-methoxybenzyl)oxy)pyridin-3-yl)-2-fluorophenyl)acetamide

[0488] 2-(4(4-ethoxy-6-((4-methoxybenzyl)oxy)pyridin-3-yl)-2-fluorophenyl)acetic acid (481 mg, 1.17 mmol) was dissolved in DMF, and then HATU (489 mg, 1.29 mmol), DMAP (14 mg, 0.117 mmol), DIEA (453 mg, 3.51 mmol), (R)-(3-amino-5-(pentafluorothio)phenyl)(3-(dimethylamino)pyrrolidin-1-yl)methanone (420 mg, 1.17 mmol) were added, and the mixture was reacted at room temperature overnight. After the reaction was complete, water was added to quench the reaction, and then EA (20 mL*2) was used for extraction. The organic phase was dried, spin-dried, and column chromatography was performed to obtain 350 mg of (R)-N-(3-(3-(dimethylamino)pyrrolidine-1-carbonyl)-5-(pentafluorothio)phenyl)-2-(4-(4-ethoxy-6-((4-methoxybenzyl)oxy)pyridin-3-yl)-2-fluorophenyl)acetamide. MS m / z (ESI): 753 [M+H] + .

[0489] Step 4 Synthesis of (R)-N-(3-(3-(dimethylamino)pyrrolidine-1-carbonyl)-5-(pentafluorothio)phenyl)-2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamide

[0490] (R)-N-(3-(3-(dimethylamino)pyrrolidine-1-carbonyl)-5-(pentafluorothio)phenyl)-2-(4-(4-ethoxy-6-((4-methoxybenzyl)oxy)pyridin-3-yl)-2-fluorophenyl)acetamide (350 mg, 0.465 mmol) was dissolved in DCM (10 mL), and then TFA (2 mL) was added, and the mixture was stirred at room temperature for 2 hours. After the reaction was complete, the reaction solution was concentrated, and then adjusted to neutral with sodium carbonate aqueous solution, extracted with DCM (20 mL*2), and the organic phase was dried by spin drying. Column chromatography was performed to obtain 200 mg of (R)-N-(3-(3-(dimethylamino)pyrrolidine-1-carbonyl)-5-(pentafluorothio)phenyl)-2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamide (Compound 13). MS m / z(ESI):633[M+H] + . 1 H NMR (400 MHz, DMSO-d 6 )δ11.40(s,1H),10.87(s,1H),8.29(t,J=2.1Hz,1H),8.04(d,J=13.0Hz,1 H),7.70(s,1H),7.41–7.33(m,2H),7.30–7.21(m,2H),5.82(s,1H),4.05(q ,J=6.9Hz,2H),3.95–3.86(m,1H),3.80(s,2H),3.73–3.47(m,4H),2.75(d ,J=50.1Hz,6H),2.29(s,1H),2.10(t,J=8.8Hz,1H),1.29(t,J=6.9Hz,3H).

[0491] Biological activity test example 1:

[0492] (a) In vitro screening experiment - HTRF method to detect the inhibitory activity of compounds on RET

[0493] Here’s how:

[0494] 1. Preparation of 1x kinase buffer: Mix 5x enzyme buffer and distilled water at a ratio of 1:4, and the final concentration is 5mM magnesium chloride and 1mM dithiothreitol.

[0495] 2. Dilute the test compound (5 mM stock solution) 5-fold to 1 mM with 100% dimethyl sulfoxide and perform 10 equal dilutions at 1:3 in a 384-well dilution plate.

[0496] 3. Use the Echo 550 pipetting system to add 0.2 μL of the gradient diluted compound to a 384-well plate, with 2 replicates for each concentration, and the final concentration of dimethyl sulfoxide is 0.5% (v / v). The gradient concentrations of the test compound are 5000, 1666.7, 555.5, 185.18, 61.72, 20.57, 6.858, 2.286, 0.764, and 0.254 nM.

[0497] 4. Prepare 2x RET (0.1 ng / μL) in 1x kinase buffer.

[0498] 5. Add 5 μL of 2x RET to the 384-well plate, centrifuge at 1000 g for 30 seconds, and incubate at room temperature for 10 minutes.

[0499] 6. Prepare a mixture of 2x tyrosine kinase-biotin-labeled substrate (2 μM) and adenosine triphosphate (20 μM) using 1x kinase buffer.

[0500] 7. Add 5 μL of tyrosine kinase-biotin-labeled substrate and adenosine triphosphate mixture to start the reaction. Centrifuge at 1000 g for 30 seconds, seal the plate, and incubate at room temperature for 30 minutes.

[0501] 8. Prepare 2x Sa-XL 665 (Note: a reagent) (125 μM) and a mixture of tyrosine kinase-antibody-cryptate using homogeneous time-resolved fluorescence detection buffer.

[0502] 9. Add 10 μL of Sa-XL 665 and tyrosine kinase-antibody-cryptate mixture to each well, centrifuge at 1000 g for 30 seconds, and incubate at room temperature for 1 hour.

[0503] 10. Read the plate at 615nm and 665nm using Envision 2104 microplate reader and calculate the ratio (665 / 615nm).

[0504] 11. The inhibition rate is calculated as follows:

[0505]

[0506] In the formula,

[0507] R0 is the average ratio of the ELISA plate in the solvent blank group

[0508] R1 is the test compound plate ratio

[0509] R2 is the average ratio of the microplate reader for 100% inhibition of RET enzyme activity

[0510] IC was calculated using GraphPad 6.0 software by fitting inhibition percentage values ​​and logarithms of compound concentrations to nonlinear regression (dose response-variable slope).50 .

[0511] (b) In vitro screening experiment - HTRF method to detect the inhibitory activity of compounds on VEGFR2

[0512] Experimental steps:

[0513] 1. Preparation of 1x kinase buffer: Mix 5x enzyme buffer and distilled water at a ratio of 1:4, and the final concentration is 5mM magnesium chloride; 1mM dithiothreitol; 1mM manganese chloride.

[0514] 2. Dilute the test compound (5 mM stock solution) 5-fold to 1 mM with 100% dimethyl sulfoxide and perform 10 equal dilutions at 1:3 in a 384-well dilution plate.

[0515] 3. Use Echo 550 pipetting system to add 0.2 μL of the serially diluted compound to a 384-well cell culture plate (Corning, 3570), with 2 replicates for each concentration, and the final concentration of dimethyl sulfoxide is 0.5% (v / v). The gradient concentration of the test compound is 5000, 1666.7, 555.5, 185.18, 61.72, 20.57, 6.858, 2.286, 0.764, 0.254 nM.

[0516] 4. Prepare 2x VEGFR2 (0.02 ng / μL) in 1x kinase buffer.

[0517] 5. Add 5 μL of 2x VEGFR2 to the 384-well plate, centrifuge at 1000 g for 30 seconds, and incubate at room temperature for 10 minutes.

[0518] 6. Prepare a 2x tyrosine kinase-biotin-labeled substrate (2 μM) and ATP (8 μM) mixture using 1x kinase buffer.

[0519] 7. Add 5 μL of tyrosine kinase-biotin-labeled substrate and adenosine triphosphate mixture to start the reaction. Centrifuge at 1000 g for 30 seconds, seal the plate, and incubate at room temperature for 40 minutes.

[0520] 8. Prepare 2x Sa-XL 665 (125 μM) and tyrosine kinase-antibody-cryptate mixture using homogeneous time-resolved fluorescence detection buffer.

[0521] 9. Add 10 μL of Sa-XL 665 and tyrosine kinase-antibody-cryptate mixture to each well, centrifuge at 1000 g for 30 seconds, and incubate at room temperature for 1 hour.

[0522] 10. Read the plate at 615nm and 665nm using Envision 2104 microplate reader and calculate the ratio (665 / 615nm).

[0523] 11. The inhibition rate is calculated as follows:

[0524]

[0525] In the formula,

[0526] R0 is the average ratio of the ELISA plate in the solvent blank group

[0527] R1 is the test compound plate ratio

[0528] R2 is the average ratio of the microplate reader for 100% inhibition of RET enzyme activity

[0529] 12. Calculate IC by fitting the inhibition percentage values ​​and logarithms of compound concentrations to nonlinear regression (dose response - variable slope) using GraphPad 6.0 50 .

[0530] (c) In vitro screening experiment - CellTiter-Glo luminescence assay to detect the inhibition of Ba / F3-KIF5B-RET cell viability by compounds

[0531] Experimental steps:

[0532] 1. Utilize The mammalian cell expression vector containing human KIF5B-RET cDNA was introduced into Ba / F3 cells by transfection system method, and the surviving clones were screened by puromycin and subjected to cell growth inhibition function experiment and western blotting method to verify the cell line with stable and high expression of RET.

[0533] 2. The cells were cultured in a medium containing RPMI 1640 culture medium, 10% fetal bovine serum, 1% penicillin-streptomycin, and 2 μg / mL puromycin in a cell culture incubator at 37° C. and 5% carbon dioxide.

[0534] 3. Dilute the test compound (5 mM stock solution) 2.5-fold to 2 mM with 100% dimethyl sulfoxide and perform 10 equal dilutions at 1:3 in a 384-well dilution plate.

[0535] 4. Use Echo 550 pipetting system to add 0.2 μL of the serially diluted compound to a 384-well cell culture plate (Corning, 3570), with 2 replicates for each concentration, and the final concentration of dimethyl sulfoxide is 0.5% (v / v). The serial concentrations of the test compound are 5000, 1666.7, 555.5, 185.18, 61.72, 20.57, 6.858, 2.286, 0.764, 0.254 nM.

[0536] 5. Add 40 μL of 800 Ba / F3-KIF5B-RET cell suspension to each well and culture in a 5% carbon dioxide cell culture incubator for 72 h.

[0537] 6. Add 20 μL of Cell Titer-Glo reagent to each well of the cell culture plate, shake and mix for 2 minutes to lyse the cells, then incubate at room temperature for 30 minutes, and read the fluorescence signal value using Envision 2104 microplate reader.

[0538] 7. Data from XLFit 5.0 according to the 4-parameter formula:

[0539] Y=Bottom+(Top-Bottom) / (1+10^((LogIC50-X)*HillSlope)) Fitting calculation IC 50 value.

[0540] (d) In vitro screening experiment - CellTiter-Glo luminescence assay to detect the inhibition of TT (human medullary thyroid carcinoma) cell viability by compounds

[0541] Experimental steps:

[0542] 1. The cells were cultured in a 384-well cell culture plate (Corning, 3570) in a medium containing F12K medium, 20% fetal bovine serum, and 1% penicillin-streptomycin in a 37°C, 5% CO2 cell culture incubator overnight.

[0543] 2. Dilute the test compound (5 mM stock) 2.5-fold to 2 mM with 100% DMSO and perform 10 equal dilutions at 1:3 in a 384-well dilution plate.

[0544] 3. Use Echo 550 to add 0.2 μL of the gradient diluted compound to 384-well cells, with 2 replicates for each concentration, and the final DMSO concentration is 0.5% (v / v). The gradient concentration of the test compound is 1000, 333.33, 111.11, 37.04, 12.35, 4.12, 1.37, 0.46, 0.15, 0.05 nM. Culture in a 5% carbon dioxide cell culture incubator for 72 hours.

[0545] 4. Add 30 μL Cell Titer-Glo Reagent to each well of the cell culture plate, shake and mix for 2 minutes to lyse the cells, then incubate at room temperature in the dark for 30 minutes, and read the luminescence signal value using Envision 2104.

[0546] 5. Data were obtained from XLFit 5.0 using the 4-parameter formula: Y = Bottom + (Top-Bottom) / (1 + 10^((LogIC50 -X)*HillSlope))Fitting calculation IC 50 value.

[0547] The test results are shown in Table 1

[0548] Table 1

[0549]

[0550]

[0551] '-' means not provided.

[0552] (e) In vitro screening experiment - HTRF method to detect the inhibitory activity of compounds on TRK

[0553] Here’s how:

[0554] 1. Preparation of 1x kinase buffer: Mix 5x enzyme buffer and distilled water at a ratio of 1:4, and the final concentration is 5mM magnesium chloride and 1mM dithiothreitol.

[0555] 2. Dilute the test compound (5 mM stock solution) 5-fold to 1 mM with 100% dimethyl sulfoxide and perform 10 equal dilutions at 1:3 in a 384-well dilution plate.

[0556] 3. Use the Echo 550 pipetting system to add 0.2 μL of the gradient diluted compound to a 384-well plate, with 2 replicates for each concentration, and the final concentration of dimethyl sulfoxide is 0.5% (v / v). The gradient concentrations of the test compound are 5000, 1666.7, 555.5, 185.18, 61.72, 20.57, 6.858, 2.286, 0.764, and 0.254 nM.

[0557] 4. Prepare 2x TRK A / TRK B / TRK C / TRK A G595R / TRK A G667C with 1x kinase buffer.

[0558] 5. Add 5 μL of 2x TRK A / TRK B / TRK C / TRK AG595R / TRK A G667C to the 384-well plate, centrifuge at 1000 g for 30 seconds, and incubate at room temperature for 10 minutes.

[0559] 6. Prepare 2x threonine protein phosphatase-biotin labeled substrate and adenosine triphosphate mixture in 1x kinase buffer.

[0560] 7. Add 5 μL of tyrosine kinase-biotin-labeled substrate and adenosine triphosphate mixture to start the reaction. Centrifuge at 1000 g for 30 seconds, seal the plate, and incubate at room temperature for 40 minutes.

[0561] 8. Prepare 4x Sa-XL 665 (Note: a reagent) and tyrosine kinase-antibody-cryptate mixture using homogeneous time-resolved fluorescence detection buffer.

[0562] 9. Add 5 μL Sa-XL 665 and 5 μL threonine protein phosphatase-antibody-cryptate mixture to each well, centrifuge at 1000 g for 30 seconds, and incubate at room temperature for 1 hour.

[0563] 10. Read the plate at 615nm and 665nm using Envision 2104 microplate reader and calculate the ratio (665 / 615nm).

[0564] 11. The inhibition rate is calculated as follows:

[0565]

[0566] In the formula,

[0567] R0 is the average ratio of the ELISA plate in the solvent blank group

[0568] R1 is the test compound plate ratio

[0569] R2 is the average ratio of the microplate reader for 100% inhibition of RET enzyme activity

[0570] IC50 was calculated using GraphPad 6.0 software by fitting the inhibition rate values ​​and logarithms of compound concentrations to nonlinear regression (dose response-variable slope). IC50 values ​​were calculated by XLFit 5.0 using the 4-parameter formula: Y = Bottom + (Top-Bottom) / (1 + 10^((LogIC50-X)*HillSlope)).

[0571] The test results are shown in Table 2:

[0572] Table 2

[0573]

[0574]

[0575] '-' means not provided.

[0576] It can be seen from the above data that the compounds listed in the present invention have good inhibitory activity against Trk A, Trk B, Trk C, and some mutants thereof.

[0577] (f) In vivo efficacy evaluation - the effect of the compound in a telemetric irritable bowel syndrome (IBS) model

[0578] Experimental animals: Normal SD rats, 280-350 g;

[0579] Test compounds: Compound 1 (10 mg / kg), BOS-589 (10 mg / kg);

[0580] Experimental steps:

[0581] The animals were anesthetized with 1.50 mg / kg sodium pentobarbital, and a DSI telemetry implant was placed in the external oblique muscle to record EMG signals;

[0582] 2. The day of drug administration was defined as the first day of the experiment (D1). From D1 to D3, drugs were administered orally twice a day. On D3.5, rectal dilation experiments were performed 1 hour after administration of drugs in the morning and rectal acetic acid stimulation. The balloon pressure was set to 60 mmHg, and EMG signals were recorded. The detection lasted for 10 minutes.

[0583] Data analysis: The raw data were collected by DSI system Ponemah software and analyzed by NeuroScore software and Spike2. The recorded EMG signals were calculated, and each pressure value was counted for 10 minutes. The experimental data were represented by the mean value (Mean), and the t-test was used for statistical analysis. P<0.05 indicated statistical significance, P<0.01 indicated significant difference, and P<0.001 indicated extremely significant difference.

[0584] Experimental results:

[0585]

[0586] The results of this experiment showed that in the rectal dilation experiment after administration, the burst number of EMG signals of the external oblique muscle of the compound 1 (10 mg / kg) group was significantly reduced, indicating that compound 1 has a certain therapeutic effect on the acetic acid-induced visceral sensitivity model of rats, and the therapeutic effect of compound 1 at the same dose is more significant than that of BOS-589.

[0587] The structure of BOS 589 is as follows:

[0588]

[0589] The structure of LOXO-195 is as follows:

[0590] The structure of GSK-3179106 is as follows:

[0591] In summary, the compounds of the present invention have good RET kinase inhibitory activity and TRK kinase inhibitory activity; the inventors also unexpectedly discovered through research that BOS 589 and GSK-3179106 also have certain TRK kinase inhibitory activity.

[0592] Compared with the compound BOS 589, the Ba / F3-KIF5B-RET cell activity of the compound of the present invention is increased by 6 times, and the TT cell activity is increased by 12 times; compared with LOXO-195, the compound of the present invention has stronger inhibitory activity against mutant TRK kinases (such as TRK AG595R / TRK A G667C); the therapeutic effect of the compound of the present invention in the IBS model (acetic acid-induced intestinal hypersensitivity model) is significantly better than that of BOS-589 at the same dose (the experimental results of some compounds are not shown). The above-mentioned unexpected technical effects of the compounds claimed for protection in the present invention are speculated by the inventors to be due to the fact that the sulfur pentafluoride functional group in the compound gives it unique physical and chemical properties and three-dimensional spatial structure, which can better bind to the corresponding pockets of RET / TRK protein, so that the compound of the present invention has better RET / TRK kinase inhibitory activity.

[0593] All documents mentioned in the present invention are cited as references in this application, just as each document is cited as reference individually. In addition, it should be understood that after reading the above teachings of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the claims attached to this application.

Claims

1. A compound of formula I or a pharmaceutically acceptable salt or isotope thereof; In the formula, R1 is R' is selected from C1-C6 alkyl; R2 is halogen; R5 is R 10 , R' 10 , R 11 , R'1 11 and R” 11 Each independently is H or C1-C6 alkyl; or R 10 and R' 10 Together with the nitrogen atom to which it is attached, it forms a substituted or unsubstituted 3-8 membered heterocyclic group, wherein: The substitution refers to the substitution of an N(R' 13 )(R” 13 ) replaces; R' 13 and R” 13 Each is independently selected from H or C1-C6 alkyl; L2 is selected from: Among them, R f and R g Each is independently selected from H, C1-C6 alkyl, halogen; r is 1; m is 1, 2, 3, 4, 5 or 6; n is 1; n' is 1; n” is 0.

2. The compound of formula I or its pharmaceutically acceptable salt or isotope compound according to claim 1, characterized in that: It has the structure shown in Formula I': In the formula, R1 is selected from C1-C6 alkoxy; R2 is selected from F; R3 and R4 are each independently selected from H; R5 is selected from R 10 , R' 10 , R 11 , R' 11 and R” 11 Each independently selected from H or C1-C6 alkyl; or R 10 and R' 10 Together with the N atom to which it is connected, it forms a substituted or unsubstituted 3-8 membered heterocyclic group, wherein the substitution refers to the substitution by one N(R' 13 )(R” 13 ) replaces; R' 13 and R” 13 Each is independently selected from H or C1-C6 alkyl; L2 is Among them, the R f and R g are each independently selected from H; n is 1; m is 1, 2 or 3.

3. The compound of formula I or its pharmaceutically acceptable salt or isotope compound according to claim 1, characterized in that: It has the structure shown in Formula I": In the formula, Rc is selected from C1-C6 alkyl; X is selected from halogen; R5 is selected from R 10 , R' 10 , R 11 , R' 11 and R” 11 Each independently is H or C1-C6 alkyl; or R 10 and R' 10 Together with the N atom to which it is connected, it forms a substituted or unsubstituted 3-8 membered heterocyclic group, wherein the substitution refers to the substitution by one N(R' 13 )(R” 13 ) replaces; R' 13 and R” 13 Each is independently selected from H or C1-C6 alkyl; L2 is Among them, the R f and R g Each is independently selected from H, C1-C6 alkyl, halogen; m is 1, 2 or 3.

4. The compound according to claim 3 or its pharmaceutically acceptable salt or isotope compound, characterized in that: X is F; R f and R g is H; m is 2.

5. The compound according to any one of claims 1 to 4 or a pharmaceutically acceptable salt or isotope compound thereof, characterized in that: R5 is selected from R 10 , R' 10 , R 11 , R' 11 and R” 11 Each independently selected from H or C1-C6 alkyl; or R 10 and R' 10 Together with the N atom to which it is attached, it forms a substituted 3-8 membered heterocyclic group, wherein the substitution refers to the substitution by one N(R' 13 )(R” 13 ) replaces; R' 13 and R” 13 Each is independently selected from H, C1-C6 alkyl.

6. The compound according to any one of claims 1 to 4 or a pharmaceutically acceptable salt or isotope compound thereof, characterized in that: R5 is selected from R 11 , R' 11 and R” 11 Each independently selected from H or C1-C6 alkyl; R 10 and R' 10 Together with the N atom to which it is connected, it forms a substituted or unsubstituted 3-8 membered heterocyclic group, wherein the substitution refers to the substitution by one N(R' 13 )(R” 13 ) replaces; R' 13 and R” 13 Each is independently selected from H, C1-C6 alkyl.

7. The compound according to claim 1 or its pharmaceutically acceptable salt or isotope compound, characterized in that: The compound is selected from 8. A pharmaceutical composition, characterized in that It comprises the compound according to any one of claims 1 to 7 or a pharmaceutically acceptable salt or isotope compound thereof; and a pharmaceutically acceptable carrier or diluent.

9. The pharmaceutical composition according to claim 8, characterized in that It also includes one or more therapeutic agents selected from the following group: aminosalicylic acid preparations, glucocorticoids, PD-1 inhibitors, PD-L1 inhibitors, CD20 antibodies, CD47 antibodies, ALK inhibitors, PI3K inhibitors, BTK inhibitors, EGFR inhibitors, VEGFR inhibitors, HDAC inhibitors, CDK inhibitors, MEK inhibitors, Akt inhibitors, mTOR inhibitors, SHP2 inhibitors, and IGF-1R inhibitors.

10. The pharmaceutical composition according to claim 9, characterized in that The aminosalicylic acid preparation is sulfasalazine; The glucocorticoid is selected from hydrocortisone or dexamethasone; The PD-1 inhibitor is selected from nivolumab, pembrolizumab, JS-001, SHR-120, BGB-A317, IBI-308, GLS-010, GB-226, STW204, HX008, HLX10, BAT1306, AK105, LZM 009; The PD-L1 inhibitor is selected from durvalumab, atezolizumab, CS1001, KN035, HLX20, SHR-1316, BGB-A333, JS003, CS1003, KL-A167, F 520, GR1405, and MSB2311; The CD20 antibody is selected from rituximab, obinutuzumab, ofatumumab, tositumomab or ibritumomab tiuxetan; The CD47 antibody is selected from Hu5F9-G4, CC-90002, TTI-621, TTI-622, OSE-172, SRF-231, ALX-148, NI-1701, SHR-1603, IBI188 or IMM01; The ALK inhibitor is selected from ceritinib, alectinib, brigatinib, lorlatinib or oclatinib; The PI3K inhibitor is selected from Idelalisib, Dactolisib, Taselisib or Buparlisib; The BTK inhibitor is selected from Ibrutinib, Tirabrutinib or Acalabrutinib; The EGFR inhibitor is selected from afatinib, gefitinib, erlotinib, lapatinib, dacomitinib, icotinib or canertinib; The VEGFR inhibitor is selected from sorafenib, pazopanib, rivatinib, cabozantinib, sunitinib or donafenib; The HDAC inhibitor is selected from Givinostat, Droxinostat, Entinostat, Dacilast or Tecodinaeline; The CDK inhibitor is selected from Palbociclib, Ribociclib, Abemaciclib or Lerociclib; The MEK inhibitor is selected from selumetinib, trametinib, PD0325901, U0126, AS-703026 or PD184352; The Akt inhibitor is selected from MK-2206, Ipatasertib, Capivasertib, Afuresertib or Uprosertib; The mTOR inhibitor is Vistusertib; The SHP2 inhibitor is selected from RMC-4630, JAB-3068 or TNO155; The IGF-1R inhibitor is selected from Ceritinib, oclatinib, linsitinib, BMS-754807 or GSK1838705A.

11. Use of the compound according to any one of claims 1 to 7 or its pharmaceutically acceptable salt, isotope compound, or the pharmaceutical composition according to any one of claims 8 to 10 in the preparation of a medicament for treating irritable bowel syndrome, other intestinal-related diseases and / or cancer; The irritable bowel syndrome, other intestinal related diseases and / or cancer are RET and / or TRK related diseases.

12. The use according to claim 11, characterized in that The irritable bowel syndrome is diarrhea-predominant irritable bowel syndrome, constipation-predominant irritable bowel syndrome, mixed irritable bowel syndrome or unclassified irritable bowel syndrome; the other intestinal-related diseases are functional bloating, functional constipation, nonspecific functional intestinal disorder, functional abdominal pain syndrome, chronic idiopathic constipation or functional gastroduodenopathy.

13. The use according to claim 11, characterized in that The cancer is bladder cancer, ovarian cancer, adenocarcinoma, gastric cancer, colon cancer, lung cancer, bone cancer, brain cancer, neuroblastoma, rectal cancer, familial adenomatous polyposis cancer, esophageal cancer, lip cancer, laryngeal cancer, hypopharyngeal cancer, tongue cancer, kidney cancer, cervical cancer, uterine body cancer, choriocarcinoma, testicular cancer, urinary cancer, melanoma, acute lymphocytic leukemia, chronic lymphocytic leukemia, acute myeloid leukemia, chronic myeloid leukemia, hepatocellular carcinoma, gallbladder cancer, bronchial cancer or multiple myeloma.

14. The use according to claim 11, characterized in that The cancer is pancreatic cancer, prostate cancer, salivary gland cancer, medullary thyroid cancer, papillary thyroid cancer, small cell lung cancer, non-small cell lung cancer, hereditary non-polyposis colorectal cancer, renal parenchymal cancer or endometrial cancer.

15. The use according to claim 11, characterized in that The cancer is non-small cell lung cancer, glioma, multiple myeloma, hepatobiliary carcinoma, medullary thyroid carcinoma, papillary thyroid tumor, neuroblastoma, colon cancer, head and neck squamous cell carcinoma, sarcoma, melanoma, pancreatic tumor, high-grade solid tumor, breast tumor or bile duct cancer.

16. The use according to claim 11, characterized in that The cancer is a fibrosarcoma or a soft tissue sarcoma.

17. Use of the compound according to any one of claims 1 to 7 or its pharmaceutically acceptable salt, isotope compound or the pharmaceutical composition according to any one of claims 8 to 10 in the preparation of a medicament for treating neurodegenerative diseases, chronic pain, acute pain, inflammatory diseases, Trypanosoma cruzi infection or diseases related to imbalanced regulation of bone remodeling; The neurodegenerative disease, chronic pain, acute pain, inflammatory disease, Trypanosoma cruzi infection or disease associated with imbalanced regulation of bone remodeling is a RET and / or TRK related disease.

18. The use according to claim 17, characterized in that The inflammatory diseases include inflammatory bowel disease.

19. Use of a compound as claimed in any one of claims 1 to 7 or a pharmaceutically acceptable salt or isotope compound thereof, or a pharmaceutical composition as claimed in any one of claims 8 to 10 in the preparation of a medicament for inhibiting RET kinase and / or TRK kinase activity in a cell or a subject.

20. Use of BOS-589 or a pharmaceutically acceptable salt or isotope compound thereof in the preparation of a drug for inhibiting TRK kinase activity in a cell or a subject, wherein the structure of BOS-589 is as follows:

21. A method for preparing the compound according to claim 1 or its pharmaceutically acceptable salt or isotope compound, characterized in that: Includes steps: 1) reacting a compound of formula M with a compound of formula N to obtain a compound of formula A; 2) reacting the compound of formula A with an acid to obtain a compound of formula I; wherein Y is selected from OH, halogen; The other groups are as defined in claim 1.

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