3-(AMINO)methylene indoline derivatives, preparation method therefor and use thereof

Abstract

Disclosed in the present invention are new 3-(amino)methylene indoline derivatives, a preparation method therefor and the pharmaceutical use thereof. More specifically, the compounds can be used as a drug for treating proliferative disorders and other diseases associated with the abnormal expression of various kinases. In particular, the compounds can be used as a drug for treating proliferative disorders and other diseases related to the abnormal expression of various kinases such as RET, PDGFR and VEGFR. The 3-(amino)methylene indoline derivatives of the present invention have a structure as shown in general formula (I), wherein R1, R2, R3, R4, R5, R6, R7, R8, L1, L2, X and Y are as defined in the description.

Description

3-(Amino)methyleneindoline derivatives and their preparation method and application Technical Field

[0001] The present invention relates to novel 3-(amino)methyleneindoline derivatives, their preparation methods, and uses. Specifically, the present invention relates to novel 3-aminoindoline derivatives that inhibit the growth of wild-type tumor strains with high kinase expression or tumor cell lines with corresponding kinase mutations, and their preparation methods. In particular, these compounds are useful as drugs for treating proliferative disorders and other diseases associated with abnormal expression of various kinases, such as RET, PDGFR, and VEGFR. Furthermore, the present invention relates to medicaments containing these compounds and their use in the preparation of medicaments. Background Art

[0002] Cancer is a major threat to human health. Chemotherapy remains an important treatment modality. Small molecule targeted therapies, particularly those used in combination with other approaches, are playing an increasingly important role. Among the numerous targeted therapeutics, kinase inhibitors, particularly those targeting RET, PDGFR (PDGFRα and PDGFRβ), and VEGFR, have garnered significant attention.

[0003] RET is a neuronal growth factor receptor tyrosine kinase and a transmembrane glycoprotein. Expressed by the proto-oncogene RET (Rearranged durIng TransfectIon) located on chromosome 10, RET plays an important role in the embryonic development of the kidney and enteric nervous system. It is also crucial for the homeostasis of various tissues, including neurons, neuroendocrine tissue, hematopoietic tissue, and male germ cells. Unlike other RTKs, RET does not directly bind to ligand molecules such as artemin, glial cell line-derived neurotrophic factor (GDNF), neurturin, and persephin, which belong to the GNDF family of ligands (GFLs). These ligands GFLs usually bind to the GDNF family receptor α (GFRα). The formed GFLs-GFRα complex mediates the self-dimerization of RET protein, causing trans-autophosphorylation of tyrosine on the intracellular domain, recruiting related adaptor proteins, and activating cascade reactions of signal transduction such as cell proliferation. Related signaling pathways include MAPK, PI3K, JAK-STAT, PKA, PKC, etc. ((1)Maria Grazia Borrello,Elena Ardini,Laura D Locati,Angela Greco,Lisa Licitra & Marco A PIerotti(2013).RET Inhibition:ImplicatIons in cancer therapy.Expert Opinion on Therapeutic Targets,17:4,403-419)(2)Samuel A.Wells,Jr,Furio Pacini,Bruce G. Robinson, and Massimo Santoro. Multiple Endocrine Neoplasia Type 2 and Familiai Meduliary Thyroid Carcinoma: An Update. J Clin Endocrinol Metab 98:3149–3164, 2013).

[0004] There are two primary mechanisms for oncogenic activation of RET: 1. Chromosomal rearrangements generate new fusion proteins, typically fusions of the RET kinase domain with a protein containing the self-dimerization domain; 2. RET mutations directly or indirectly activate RET kinase activity. These alterations at the somatic or germline level have been implicated in the pathogenesis of various cancers. RET chromosomal rearrangements are found in 5%-10% of patients with papillary thyroid carcinoma; RET point mutations are found in 60% of patients with medullary thyroid carcinoma. Approximately 1-2% of all NSCLC patients harbor RET fusion proteins, with KIF5B-RET being the most common.

[0005] These facts indicate that it is an ideal treatment for tumors related to persistent RET activation. The research on RET inhibitors has received widespread attention and has made rapid progress. Among them, RET inhibitors represented by Pralsetinib (WO2017011776A1Array Loxo 292) and Selpercatinib (WO2017079140A1Blu 667) have been approved by the FDA for marketing and have been used to treat various related fusion tumors regardless of tumor type and have achieved success. This has inspired various other development attempts. ((1) Lucille Lopez-Delisle, CécIle Pierre-Eugène, Caroline Louis-Brennetot, Didier Surdez, Virginie Raynal, Sylvain Baulande, Valentina Boeva, Sandrine Valérie Combaret,Michel Peuchmaur6,Olivier Delattre,Isabelle Janoueix-Lerosey.Activated ALK signals through the ERK–ETV5–RET pathway to drive neuroblastoma oncogenesis.Oncogene(2018)37:1417–1429.(2)WO 2014 / 141187A1.RET Kinase Inhibitors May Treat Cancer and Gastrointestinal Disorders.(3)MInsoo Song.Progress In Discovery of KIF5B-RET Kinase Inhibitors for the Treatment ofNon-Small-Cell Lung Cancer.Miniperspective.J.Med.Chem.2015,58,3672-3681.)

[0006] Platelet-derived growth factor (PDGF) is a family of effective mitogens for almost all mesenchyme-derived cells. PDGF exerts its cellular effects through PDGF receptor α (PDGFRα) and PDGF receptor β (PDGFRβ). PDGFRα is structurally similar to C1-C8 and can form heterodimers and homodimers. Currently reported examples of inhibitors for both PDGFRα and PDGFRβ include Nintedanib and the like. In addition to their inhibitory effects on PDGFRα and PDGFRβ, they also have inhibitory effects on cKIT, BCR-ABL, etc. (An overview of kinase downregulators and recent advances in discovery approaches.Beilei Wang,Hong Wu,Chen Hu,Haizhen Wang,Jing Liu,Wenchao Wang,and Qingsong Liu.Signal Transduction and Targeted Therapy(2021)6:423)

[0007] Vascular endothelial growth factors (VEGFs), originally discovered as vascular permeability factors (VPFs), are cytokines secreted by tumor cells that increase vascular permeability. Current research has shown that VEGFs are hormonal regulators of endothelial cell differentiation. VEGFs bind to VEGFR (vascular endothelial growth factor receptor) tyrosine kinases, which are specifically and highly expressed on the surface of newly formed endothelial cells, activating the tyrosine kinases and thus exerting their biological functions. Studies have shown that VEGFR inhibitors can block VEGFR in vitro, thereby inhibiting the binding of VEGF to VEGFR. Related kinase inhibitors, such as lenvatinib mesylate, have been approved by the FDA for marketing and are clinically used to treat renal cancer, liver cancer, and thyroid cancer (Overview of lenvatinib as a targeted therapy for advanced hepatocellular carcinoma. Obaid Rehman, Urooj Jaferi, Inderbir Padda, Nimrat Khehra, Harshan Atwal, Dina Mossabeh, Ranvir Bhangu. Clin Exp HEPATOL 2021; 7, 3: 249–257).

[0008] As mentioned above, the research and development of kinase inhibitors targeting RET has achieved great success in the treatment of fusion tumors (Alexander Drilon, Zishuo I. Hu, Gillianne GYLai & Daniel SW Tan Targeting RET-driven cancers: lessons from evolving preclinical and clinical landscapes. Nature Reviews Clinical Oncology volume 15, 2018, 151–167). However, if it is possible to simultaneously inhibit multiple kinases such as PDGFR, VEGFR, FGFR, FLT3, Aurora-A, Aurora-B, TRK, B-RAF, RET or Abl while inhibiting RET, it will potentially help solve the problem of drug resistance caused by related drugs. Therefore, this application intends to study multi-target kinase inhibitors for kinases such as RET, PDGFR, and VEGFR to meet the huge social needs. Summary of the Invention

[0009] One objective of the present invention is to disclose novel 3-(amino)methyleneindoline derivatives. These compounds interact with multiple kinases, including RET, PDEGR, and VEGFR, and can be used to treat tumors, endocrine disorders, immune system diseases, genetic diseases, and neurodegenerative diseases.

[0010] The second object of the present invention is to disclose a method for preparing the 3-(amino)methyleneindoline derivatives.

[0011] More specifically, these compounds can be used as drugs for treating proliferative disorders and other diseases associated with abnormal expression of various kinases. In particular, these compounds can be used as drugs for treating proliferative disorders and other diseases associated with abnormal expression of various kinases such as RET, PDGFR, and VEGFR. The structure of the novel pyrimidines of the present invention is shown in the general formula (I), wherein R 1 、R 2 、R 3 、R 4 、R 5 、R 6 、R 7 、R 8 , L 1 , L 2 , X, and Y are defined as follows:

[0012] In the above general formula (I),

[0013] R 1 Selected from: hydrogen atom, alkyl, heteroalkyl, arylalkyl, C3-C14 cycloalkyl, aryl, heteroaryl, heteroarylalkyl, heterocycloalkyl, carboxylalkyl, carboxylcycloalkyl; any of the above groups independently may be unsubstituted or substituted by one or more substituents, these substituents include but are not limited to halogen, isotope, amino, carboxyl, phenyl, benzyl, phenyloxy, =O, -CF3, haloalkyl, alkyl, alkenyl, alkynyl, hydroxyl, hydroxyalkyl, alkoxy, alkoxyalkyl, heteroalkyl, arylalkyl, cycloalkyl, aryl, heteroaryl, heteroarylalkyl, heterocycloalkyl, heterocycloalkenyl, alkoxyalkyl, alkenyloxy, alkynyloxy, amino, alkylamino, aminoalkyl, alkylaminocarbonyl, sulfonyl, alkylsulfonyl, alkylsulfinyl, or aminosulfonyl; the alkyl group is a C1-C8 straight chain or branched alkyl group;

[0014] R 2Selected from: hydroxy, alkyl, aryl, alkoxy, heteroalkoxy, heteroalkoxy, arylalkoxy, C3-C8 cycloalkoxy, aryloxy, heteroaryloxy, heteroarylalkoxy, heterocycloalkoxy, amino, alkylamino, heteroalkylamino, arylalkylamino, cycloalkylamino, arylamino, heteroarylamino, heteroarylalkylamino, heterocycloalkylamino; any of the above groups may be independently unsubstituted or may be substituted with one or more substituents, including but not limited to halogen, isotope, amino, carboxyl, phenyl, benzyl, phenyloxy, =O, -CF3, haloalkyl, alkyl, alkenyl, alkynyl, hydroxy, hydroxyalkyl, alkoxy, alkoxyalkyl, heteroalkyl, arylalkyl, cycloalkyl, aryl, heteroaryl, heteroarylalkyl, heterocycloalkyl, heterocycloalkenyl, alkoxyalkyl, alkenyloxy, alkynyloxy, amino, alkylamino, aminoalkyl, alkylaminocarbonyl, sulfonyl, alkylsulfonyl, alkylsulfinyl, or aminosulfonyl; the alkyl group is a C1-C8 straight chain or branched alkyl group;

[0015] R 3 、R 4 Each of the following groups is independently selected from the group consisting of: hydrogen, halogen, isotope, hydroxyl, amino, carboxyl, alkyl, heteroalkyl, alkenyl, alkynyl, arylalkyl, C3-C14 cycloalkyl, aryl, heteroaryl, heteroarylalkyl, heterocycloalkyl, alkyloxy, heteroalkyloxy, arylalkoxy, cycloalkyloxy, aryloxy, heteroaryloxy, heteroarylalkoxy, heterocycloalkyloxy, amino, alkylamino, heteroalkylamino, arylalkylamino, cycloalkylamino, arylamino, heteroarylamino, heteroarylalkylamino, heterocycloalkylamino, carboxyl, carboxylalkylaminocarbonyl, cycloalkylaminocarbonyl, heteroarylaminocarbonyl, heteroarylaminocarbonyl; any of the above groups Each group may be unsubstituted or substituted with one or more substituents, including but not limited to halogen, isotope, amino, carboxyl, phenyl, benzyl, phenyloxy, =O, -CF3, haloalkyl, alkyl, alkenyl, alkynyl, hydroxyl, hydroxyalkyl, alkoxy, alkoxyalkyl, heteroalkyl, arylalkyl, cycloalkyl, aryl, heteroaryl, heteroarylalkyl, heterocycloalkyl, heterocycloalkenyl, alkoxyalkyl, alkenyloxy, alkynyloxy, alkylamino, aminoalkyl, alkylaminocarbonyl, sulfonyl, alkylsulfonyl, alkylsulfinyl, or aminosulfonyl; the alkyl group is a C1-C8 straight chain or branched alkyl group;

[0016] R 5Selected from: hydrogen atom, isotope, alkyl, heteroalkyl, arylalkyl, C3-C14 cycloalkyl, aryl, heteroaryl, heteroarylalkyl, heterocycloalkyl; any of the above groups independently may be unsubstituted or substituted by one or more substituents, these substituents include but are not limited to halogen, isotope, amino, carboxyl, phenyl, benzyl, phenyloxy, =O, -CF3, haloalkyl, alkyl, alkenyl, alkynyl, hydroxyl, hydroxyalkyl, alkoxy, alkoxyalkyl, heteroalkyl, arylalkyl, cycloalkyl, aryl, heteroaryl, heteroarylalkyl, heterocycloalkyl, heterocycloalkenyl, alkoxyalkyl, alkenyloxy, alkynyloxy, alkylamino, aminoalkyl, alkylaminocarbonyl, sulfonyl, alkylsulfonyl, alkylsulfinyl, or aminosulfonyl; the alkyl group is a C1-C8 straight chain or branched alkyl group;

[0017] R 6 Selected from: hydrogen atom, alkyl, heteroalkyl, arylalkyl, C3-C14 cycloalkyl, heteroarylalkyl, heterocycloalkyl; any of the above groups independently may be unsubstituted or substituted with one or more substituents, including alkyl and alkoxy; the alkyl group is a C1-C8 straight chain or branched alkyl group;

[0018] R 7 Selected from: absence, hydrogen atom, isotope, halogen, alkyl, heteroalkyl, arylalkyl, C3-C14 cycloalkyl, aryl, heteroaryl, heteroarylalkyl, heterocycloalkyl; alkoxy, heteroalkoxy, arylalkoxy, cycloalkyloxy, aryloxy, heteroaryloxy, heteroarylalkoxy, heterocycloalkyloxy, amino, alkylamino, heteroalkylamino, arylalkylamino, cycloalkylamino, arylamino, heteroarylamino, heteroarylalkylamino, heterocycloalkylamino, carboxyl, alkylaminocarbonyl, any of the above groups independently may be unsubstituted or substituted with one or more substituents, including but not limited to halogen, alkyl, alkoxy; the alkyl group is a C1-C8 straight chain or branched alkyl group;

[0019] R 7 Can be linked to the C on the benzimidazole ring 4 -、C 5 -、C 6 - or C 7 -position;

[0020] L 1Selected from: covalent bond, alkylene, heteroalkylene, -C(O)-, -C(O)-C(O)-, -C(O)-NH-, alkylene-NH-, alkylene-C(O)-NH-, -C(O)-alkylene, -C(O)-NH-heteroalkylene, heteroalkylene-NH-C(O)-, heteroalkylene-NH-; the alkylene is a C1-C8 straight chain or branched alkyl; any of the above groups may be independently substituted by one or more substituents, including hydrogen atoms and alkyl groups;

[0021] L 2 Selected from: hydrogen, alkyl, C3-C14 cycloalkyl, aryl, heteroaryl, heteroarylalkyl, heterocycloalkyl, alkoxy, heteroalkoxy, arylalkoxy, cycloalkoxy, aryloxy, heteroaryloxy, heteroarylalkoxy, heterocycloalkoxy; any of the above groups independently may be unsubstituted or substituted by one or more substituents, including but not limited to halogen, isotope, amino, carboxyl, phenyl, benzyl, phenyloxy, =O, -CF3, haloalkyl, alkyl, alkenyl, alkynyl, hydroxyl, hydroxyalkyl, alkoxy, alkoxyalkyl, heteroalkyl, arylalkyl, cycloalkyl, aryl; the alkyl group is a C1-C8 straight chain or branched alkyl group;

[0022] R 8 Selected from: absence, hydrogen atom, deuterium atom, hydroxyl, alkyl, hydroxyalkyl, heteroalkyl, alkenyl, alkynyl, arylalkyl, C3-C14 cycloalkyl, aryl, heteroaryl, arylalkyl, heteroarylalkyl, heterocycloalkyl, alkoxy, heteroalkoxy, arylalkoxy, cycloalkoxy, aryloxy, heterocycloalkoxy; any of the above groups independently may be unsubstituted or substituted with one or more substituents, including but not limited to halogen, isotope, amino, carboxyl, phenyl, benzyl, phenyloxy, =O, -CF3, haloalkyl, alkyl, alkenyl, alkynyl, hydroxyl, hydroxyalkyl, alkoxy, alkoxyalkyl, heteroalkyl, arylalkyl, cycloalkyl; the alkyl group is a C1-C8 straight chain or branched alkyl group;

[0023] X is selected from: -CHR 9 -、-N(R 10 )-、-C(O)-;

[0024] Y is selected from: a carbon atom, a nitrogen atom;

[0025] R 9 and R 10Each independently: a hydrogen atom, a deuterium atom, an alkyl group, a heteroalkyl group, a C3-C14 cycloalkyl group; any of the above groups may be unsubstituted or substituted with one or more substituents, including but not limited to halogens, isotopes, halogenated alkyl groups, and alkyl groups; the alkyl group is a C1-C8 straight chain or branched chain alkyl group;

[0026] (R 6 )N can be linked to the C of the indoline ring 5 - or C 6 - position, in which case the general formula (I) is selected from:

[0027] In certain embodiments, according to the above general formula (I), R 1 Selected from hydrogen atom, alkyl, heteroalkyl, arylalkyl, cycloalkyl, heteroarylalkyl, heterocycloalkyl, alkoxyalkyl, aminoalkyl, alkylsulfonyl, alkylsulfinyl; in the above groups, each may be unsubstituted or substituted by one or more substituents, and these substituents include: halogen, isotope, =O, -CF3, alkyl, alkenyl, alkynyl, hydroxyl, hydroxyalkyl, alkoxy, alkoxyalkyl.

[0028] In certain embodiments, according to the above general formula (I), R 1 Selected from C1-C5 alkyl, C3-C8 cycloalkyl.

[0029] R 2 Selected from: hydroxy, alkoxy, heteroalkoxy, heteroalkoxy, arylalkoxy, C3-C8 cycloalkoxy, aryloxy, heteroaryloxy, heteroarylalkoxy, heterocycloalkoxy, amino, alkylamino, heteroalkylamino, arylalkylamino, cycloalkylamino, arylamino, heteroarylamino, heteroarylalkylamino, heterocycloalkylamino; any of the above groups may be independently unsubstituted or substituted with one or more substituents, including but not limited to halogen, isotope, amino alkyl, alkylamino, alkylaminocarbonyl, sulfonyl, alkylsulfonyl, alkylsulfinyl, or aminosulfonyl; the alkyl group is a C1-C8 straight chain or branched chain alkyl group.

[0030] In certain embodiments, according to the above general formula (I), R 2 Selected from: methoxy, ethoxy, propoxy, isopropoxy, hydroxy, butoxy, difluoroethoxy, D3CD2CO-, D3CO-.

[0031] In certain embodiments, according to the above general formula (I), R 2 Independently selected from: hydroxy, C1-C5 straight or branched alkylalkoxy, heteroalkoxy, heteroalkoxy, arylalkoxy, C3-C8 cycloalkoxy, heteroalkylamino, arylalkylamino, cycloalkylamino, arylamino, heteroarylamino; any of the above groups can be substituted by one or more substituents, including but not limited to halogen, carboxyl, phenyl, =O, -CF3, alkyl, alkenyl, alkynyl, hydroxy, hydroxyalkyl, alkoxy, alkoxyalkyl, heteroalkyl, arylalkyl, cycloalkyl.

[0032] R 3 、R 4 Each of the following groups is independently selected from the group consisting of: hydrogen, halogen, isotope, hydroxyl, amino, carboxyl, alkyl, heteroalkyl, alkenyl, alkynyl, arylalkyl, C3-C14 cycloalkyl, aryl, heteroaryl, heteroarylalkyl, heterocycloalkyl, alkyloxy, heteroalkyloxy, arylalkoxy, cycloalkyloxy, aryloxy, heteroaryloxy, heteroarylalkoxy, heterocycloalkyloxy, amino, alkylamino, heteroalkylamino, arylalkylamino, cycloalkylamino, arylamino, heteroarylamino, heteroarylalkylamino, heterocycloalkylamino, carboxyl, carboxylalkylaminocarbonyl, cycloalkylaminocarbonyl, heteroarylaminocarbonyl, heteroarylaminocarbonyl; any of the above groups Each group may be unsubstituted or substituted with one or more substituents, including but not limited to halogen, isotope, amino, carboxyl, phenyl, benzyl, phenyloxy, =O, -CF3, haloalkyl, alkyl, alkenyl, alkynyl, hydroxy, hydroxyalkyl, alkoxy, alkoxyalkyl, heteroalkyl, arylalkyl, cycloalkyl, aryl, heteroaryl, heteroarylalkyl, heterocycloalkyl, heterocycloalkenyl, alkoxyalkyl, alkenyloxy, alkynyloxy, alkylamino, aminoalkyl, alkylaminocarbonyl, sulfonyl, alkylsulfonyl, alkylsulfinyl, or aminosulfonyl; the alkyl group is a C1-C8 straight chain or branched alkyl group.

[0033] In certain embodiments, according to the above general formula (I), R 4 Selected from: cyclopropane, cyclobutane, cyclopentane, methyl, ethyl, propyl, isopropyl, isobutyl, -CH(CH3)CH2CH3.

[0034] In certain embodiments, according to the above general formula (I), when R 3 When it is a hydrogen atom, R 4is selected from the group consisting of: hydrogen atom, halogen, isotope, alkyl, heteroalkyl, alkenyl, alkynyl, arylalkyl, C3-C14 cycloalkyl, aryl, heteroaryl, heteroarylalkyl, heterocycloalkyl, alkoxy, heteroalkoxy, arylalkoxy, cycloalkyloxy, aryloxy, heteroaryloxy, heteroarylalkoxy, heterocycloalkoxy, amino, alkylamino, heteroalkylamino, arylalkylamino, cycloalkylamino, arylamino, heteroarylamino, heteroarylalkylamino, heterocycloalkylamino, carboxyl, carboxylalkylaminocarbonyl, cycloalkylaminocarbonyl, heteroarylaminocarbonyl, heteroarylaminocarbonyl; the above radicals are Any group in the group may be independently unsubstituted or substituted with one or more substituents, and these substituents include but are not limited to halogen, isotope, amino, carboxyl, phenyl, benzyl, phenyloxy, =O, -CF3, haloalkyl, alkyl, alkenyl, alkynyl, hydroxyl, hydroxyalkyl, alkoxy, alkoxyalkyl, heteroalkyl, arylalkyl, cycloalkyl, aryl, heteroaryl, heteroarylalkyl, heterocycloalkyl, heterocycloalkenyl, alkoxyalkyl, alkenyloxy, alkynyloxy, alkylamino, aminoalkyl, alkylaminocarbonyl; the alkyl group is a C1-C8 straight chain or branched alkyl group.

[0035] In certain embodiments, according to the above general formula (I), when R 4 When it is a hydrogen atom, R 3 is selected from the group consisting of: hydrogen atom, halogen, isotope, alkyl, heteroalkyl, alkenyl, alkynyl, arylalkyl, C3-C14 cycloalkyl, aryl, heteroaryl, heteroarylalkyl, heterocycloalkyl, alkoxy, heteroalkoxy, arylalkoxy, cycloalkyloxy, aryloxy, heteroaryloxy, heteroarylalkoxy, heterocycloalkoxy, amino, alkylamino, heteroalkylamino, arylalkylamino, cycloalkylamino, arylamino, heteroarylamino, heteroarylalkylamino, heterocycloalkylamino, carboxyl, carboxylalkylaminocarbonyl, cycloalkylaminocarbonyl, heteroarylaminocarbonyl, heteroarylaminocarbonyl; the above radicals are Any group in the group may be independently unsubstituted or substituted with one or more substituents, and these substituents include but are not limited to halogen, isotope, amino, carboxyl, phenyl, benzyl, phenyloxy, =O, -CF3, haloalkyl, alkyl, alkenyl, alkynyl, hydroxyl, hydroxyalkyl, alkoxy, alkoxyalkyl, heteroalkyl, arylalkyl, cycloalkyl, aryl, heteroaryl, heteroarylalkyl, heterocycloalkyl, heterocycloalkenyl, alkoxyalkyl, alkenyloxy, alkynyloxy, alkylamino, aminoalkyl, alkylaminocarbonyl; the alkyl group is a C1-C8 straight chain or branched alkyl group.

[0036] In certain embodiments, according to the above general formula (I), R 5Selected from: hydrogen atom, isotope, alkyl, heteroalkyl, C3-C14 cycloalkyl, aryl, heteroaryl, heteroarylalkyl, heterocycloalkyl; any of the above groups independently may be unsubstituted or may be substituted by one or more substituents, and these substituents include but are not limited to halogen, isotope, =O, -CF3, haloalkyl, alkyl, alkenyl, alkynyl, alkoxy, alkoxyalkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, heterocycloalkenyl, alkoxyalkyl; the alkyl group is a C1-C8 straight chain or branched alkyl group.

[0037] In certain embodiments, according to the above general formula (I), R 5 Selected from: hydrogen atom, benzene ring, methyl group, ethyl group, propyl group.

[0038] In certain embodiments, according to the above general formula (I), R 6 Selected from: hydrogen atom, alkyl, heteroalkyl, arylalkyl, C3-C14 cycloalkyl, heteroarylalkyl, heterocycloalkyl; any of the above groups may be unsubstituted or substituted by one or more alkyl or alkoxy groups; the alkyl group is a C1-C8 straight chain or branched chain alkyl group.

[0039] In certain embodiments, according to the above general formula (I), R 7 Selected from: absence, hydrogen atom, isotope, halogen, alkyl, heteroalkyl, aryl, heteroaryl, heteroarylalkyl, heterocycloalkyl; alkoxy, heteroalkoxy, arylalkoxy, cycloalkoxy, aryloxy, heteroaryloxy, heteroarylalkoxy, heterocycloalkoxy; any of the above groups may be unsubstituted or substituted by one or more substituents, and these substituents include but are not limited to halogen, alkyl, alkoxy; the alkyl is a C1-C8 straight chain or branched alkyl.

[0040] In certain embodiments, according to the above general formula (I), R 7 For fluorine.

[0041] In certain embodiments, according to the above general formula (I), R 7 Can be linked to the C on the benzimidazole ring 4 -、C 5 -、C 6 - or C 7 -position.

[0042] In certain embodiments, according to the above general formula (I), L 1Selected from: covalent bond, alkylene, -C(O)-, -C(O)-C(O)-, -C(O)-NH-, alkylene-NH-, alkylene-C(O)-NH-, -C(O)-alkylene, -C(O)-NH-heteroalkylene, heteroalkylene-NH-C(O)-, heteroalkylene-NH-; the alkylene is a C1-C8 straight chain or branched alkyl; any of the above groups may be independently substituted by one or more substituents, and these substituents include hydrogen atoms and alkyl groups.

[0043] In certain embodiments, according to the above general formula (I), L 1 Selected from: -C(O)CH2-, -C(O)-, -C(O)-C(O)-, ethylene, methylene, propylene, -SO2-NH-, -CH2CH2NH-, -CONH-, -NH-.

[0044] In certain embodiments, according to the above general formula (I), L 2 Selected from: hydrogen, alkyl, C3-C14 cycloalkyl, aryl, heteroaryl, heteroarylalkyl, heterocycloalkyl, alkoxy, heteroalkoxy, arylalkoxy, cycloalkoxy, aryloxy, heteroaryloxy, heteroarylalkoxy, heterocycloalkoxy; any of the above groups independently may be unsubstituted or may be substituted by one or more substituents, and these substituents include but are not limited to halogen, isotope, amino, carboxyl, phenyl, benzyl, phenyloxy, =O, -CF3, haloalkyl, alkyl, alkenyl, alkynyl, hydroxyl, hydroxyalkyl, alkoxy, alkoxyalkyl, heteroalkyl, arylalkyl, cycloalkyl, aryl; the alkyl is a C1-C8 straight chain or branched alkyl.

[0045] In certain other embodiments, according to the above general formula (I), L 2 More preferably, they are independently selected from: methyl, Benzene ring, m-fluorophenyl, -CF3, -CH2CF3, ethylene, CF3, NH2, -OCH3, -CH2CH(CH3)2OH, -CH(NH)NH2, Fluorophenyl, -OCH(CH3)2, -NH(CH3)2, -CH2OHCH3,

[0046] In certain embodiments, according to the above general formula (I), R 8 Selected from: absence, hydrogen atom, deuterium atom, hydroxyl, alkyl, hydroxyalkyl, heteroalkyl, alkenyl, alkynyl, arylalkyl, C3-C14 cycloalkyl, aryl, heteroaryl, arylalkyl, heteroarylalkyl, cycloalkyl, heterocycloalkyl, alkoxy, heteroalkoxy, arylalkoxy, cycloalkoxy, aryloxy, heterocycloalkoxy; any of the above groups independently may be unsubstituted or may be substituted by one or more substituents, these substituents include but are not limited to halogen, isotope, amino, carboxyl, phenyl, benzyl, phenyloxy, =O, -CF3, haloalkyl, alkyl, alkenyl, alkynyl, hydroxyl, hydroxyalkyl, alkoxy, alkoxyalkyl, heteroalkyl, arylalkyl, cycloalkyl; the alkyl is a C1-C8 straight chain or branched alkyl.

[0047] In certain embodiments, according to the above general formula (I), R 8 It is a methyl group.

[0048] In certain embodiments, according to the above general formula (I), R 8 C 3 -disubstituted methyl group.

[0049] In certain embodiments, according to the above general formula (I), X is selected from: -CH(R 9 )-、-N(R 9 )-、-C(O)-。

[0050] In certain embodiments, according to the above general formula (I), R 9 and R 10 Each independently: a hydrogen atom, a deuterium atom, an alkyl group, a heteroalkyl group, a C3-C14 cycloalkyl group; any of the above groups may be unsubstituted or substituted by one or more substituents, and these substituents include but are not limited to halogens, isotopes, halogenated alkyl groups, and alkyl groups; the alkyl group is a C1-C8 straight chain or branched alkyl group.

[0051] In certain embodiments, according to the above general formula (I), (R 6 )N can be linked to the C of the indoline ring 5 - or C 6 - position, in which case the general formula (I) is selected from:

[0052] In certain embodiments, according to the above general formula (I), the structure of the 3-(amino)methyleneindoline derivative can be selected from one of the following structures, or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, solvates, metabolites, pharmaceutically acceptable salts or prodrugs:

[0053] The present invention also claims a method for preparing the above-mentioned compound, which is as follows:

[0054] When (R 6 )N can be linked to the C of the indoline ring 5 - position, and R 1 =R 3 =R 4 =R 6 =H, R 2 =OCH3, the preparation method of the 3-(amino)methyleneindoline derivatives comprises the following steps:

[0055] S1. reacting a substituted 5-nitro-indoline derivative (VI) with a chloro compound (VII) to obtain a 5-nitro-substituted indoline derivative (VIII);

[0056] S2, 5-nitro-substituted indoline derivative (VIII) reacts with active hydrogen-containing compound (IX) in toluene under heating to obtain compound (X);

[0057] S3. Compound (X) is reduced to obtain compound (XI), which is then reacted with 3-methoxymethylene-oxoindoline-6-carboxylate (XII) to obtain the target compound represented by formula (I), i.e., (XIII);

[0058] The synthetic route is as follows:

[0059] When (R 6 )N can be linked to the C 5 - position, and R 1 =R 3 =R 5 =R 6 =H, R 2 =OCH3 and X=-C(O)-, the preparation method of the 3-(amino)methyleneindoline derivatives comprises the following steps:

[0060] S1. Substituted 4-bromo-2-oxoindoline-6-carboxylic acid methyl ester (XIV) reacts with a boron derivative (XV) in the presence of a palladium reagent to obtain compound (XVI);

[0061] S2. Using a suitable acid as solvent, compound (XVI) reacts with triethyl orthoformate to obtain compound (XVII);

[0062] S3, 5-nitroindole derivative (XVIII) reacts with a suitable compound (XIX) under base catalysis to obtain compound (XX);

[0063] S4, compound (XX) is reduced to obtain the important intermediate (XXI);

[0064] S5. Compound (XVII) undergoes substitution reaction with intermediate (XXI) to obtain target compound (XXII) represented by general formula (I);

[0065] The synthetic route is as follows:

[0066] In some embodiments, the compounds described herein have one of the following structures, or a stereoisomer, geometric isomer, tautomer, N-oxide, solvate, metabolite, pharmaceutically acceptable salt, or prodrug thereof.

[0067] The present invention also includes a pharmaceutical composition containing the compound represented by the above-mentioned general formula (I) or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, solvates, metabolites, pharmaceutically acceptable salts or prodrugs.

[0068] The present invention also includes a pharmaceutical composition comprising the compound represented by the above-mentioned general formula (I) or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, solvates, metabolites, pharmaceutically acceptable salts, prodrugs; and used in combination with one or more other drugs.

[0069] The present invention includes any pharmaceutical dosage form formed by the compound represented by general formula (I) and a pharmaceutically acceptable diluent, excipient or carrier.

[0070] The present invention provides a method for treating diseases caused by, associated with or accompanied by the disruption of cell proliferation and / or angiogenesis by using an effective amount of a compound represented by general formula (I) alone or in combination with other drugs.

[0071] In certain embodiments, the disorder is a proliferative disease.

[0072] In certain embodiments, the proliferative disease is cancer.

[0073] The present invention also includes the use of the compound represented by the above general formula (I) or a pharmaceutically acceptable salt thereof or the above pharmaceutical composition for inhibiting the activity of kinases.

[0074] In certain embodiments, the inhibition of kinase activity is inhibition of RET activity.

[0075] The present invention also includes a method for treating a patient's condition caused by, or associated with, or accompanied by the disruption of cell proliferation and / or angiogenesis, comprising administering to the patient a therapeutically effective amount of a compound represented by the above-mentioned general formula (I) or a pharmaceutically acceptable salt thereof.

[0076] The present invention also includes a method for treating a disease that can be treated by inhibiting kinases in a patient, comprising administering to the patient a therapeutically effective amount of a compound represented by the above general formula (I) or a pharmaceutically acceptable salt thereof.

[0077] Conditions caused, associated or accompanied by cell proliferation and / or angiogenesis include but are not limited to: bone cancers, including Ewing's sarcoma, osteosarcoma, chondrosarcoma, etc.; brain and CNS tumors, including acoustic neuroma, neuroblastoma, glioma and other brain tumors, spinal cord tumors, breast cancer, colorectal cancer, advanced colorectal adenocarcinoma; endocrine cancers, including adrenal cortical carcinoma, pancreatic cancer, pituitary cancer, thyroid cancer, parathyroid cancer, thymic cancer, multiple endocrine neoplasia; gastrointestinal cancer Cancers include: gastric cancer, esophageal cancer, small intestine cancer, liver cancer, extrahepatic bile duct cancer, gastrointestinal carcinoid tumors, gallbladder cancer; genitourinary cancers include: testicular cancer, penile cancer, prostate cancer; gynecological cancers include: cervical cancer, ovarian cancer, vaginal cancer, uterine / endometrial cancer, pudendal cancer, gestational trophoblastic tumor, fallopian tube cancer, uterine sarcoma; head and neck cancers include: oral cancer, lip cancer, salivary gland cancer, laryngeal cancer, hypopharyngeal cancer, pharyngeal cancer, nasal cancer, paranasal sinus cancer, nasopharyngeal cancer; blood cancer Cancers, including: childhood leukemia, acute lymphocytic leukemia, acute myeloid leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, hairy cell leukemia, acute promyelocytic leukemia, plasma cell leukemia; bone marrow cancer and blood disorders, including: myelodysplastic syndrome, myeloproliferative disorders, aplastic anemia, Fanconi anemia, essential macroglobulinemia; lung cancers, including: small cell lung cancer, non-small cell lung cancer; lymphomas, including: Hodgkin's disease, non-Hodgkin's lymphoma, cutaneous T-cell lymphoma, peripheral T-cell lymphoma, AIDS-related lymphoma; eye cancers, including retinoblastoma and uveal melanoma; skin cancers, including melanoma, non-melanoma skin cancer, and Merkel cell carcinoma; soft tissue sarcomas, such as childhood soft tissue sarcoma, adult soft tissue sarcoma, and Kaposi's sarcoma; urinary system cancers, including renal cancer, Wilms' tumor, bladder cancer, urethral cancer, and metastatic cell carcinoma.

[0078] In certain embodiments, in the above methods, the patient is undergoing surgery or radiotherapy, and the compound is administered to the patient concomitantly with, before, or after the surgery or radiotherapy. BRIEF DESCRIPTION OF THE DRAWINGS

[0079] FIG1 shows the changes in tumor growth in mice bearing TT cell xenograft tumor models after administration of Example 1;

[0080] Figure 2 shows the weight change rate of TT cell xenograft tumor-bearing mice after administration of Example 1. DETAILED DESCRIPTION

[0081] The present invention discloses methods for preparing novel 3-(amino)methyleneindoline derivatives and their medical applications. These compounds can be used as, but not limited to, kinase inhibitors. The disclosed 3-(amino)methyleneindoline derivatives can be used alone or in combination with other drugs or pharmaceutically acceptable carriers, diluents, or excipients, and are suitable for preventing or treating conditions caused by, associated with, or associated with the disruption of cell proliferation and / or angiogenesis. One example of such conditions is cancer.

[0082] As used herein, the term "cancer" generally refers to a broad range of disorders characterized by the uncontrolled, abnormal growth of cells.

[0083] The compounds of the present invention are expected to be useful in treating various cancers, including but not limited to: bone cancers, including Ewing's sarcoma, osteosarcoma, chondrosarcoma, etc.; brain and CNS tumors, including acoustic neuroma, neuroblastoma, glioma and other brain tumors, spinal cord tumors, breast cancer, colorectal cancer, advanced colorectal adenocarcinoma; endocrine cancers, including adrenocortical carcinoma, pancreatic cancer, pituitary cancer, thyroid cancer, parathyroid cancer, thymic carcinoma, multiple endocrine neoplasia; gastrointestinal cancers, These include: gastric cancer, esophageal cancer, small intestinal cancer, liver cancer, extrahepatic bile duct cancer, gastrointestinal carcinoid tumors, and gallbladder cancer; genitourinary cancers, including: testicular cancer, penile cancer, and prostate cancer; gynecological cancers, including: cervical cancer, ovarian cancer, vaginal cancer, uterine / endometrial cancer, pudendal cancer, gestational trophoblastic tumor, fallopian tube cancer, and uterine sarcoma; head and neck cancers, including: oral cancer, lip cancer, salivary gland cancer, laryngeal cancer, hypopharyngeal cancer, pharyngeal cancer, nasal cancer, paranasal sinus cancer, and nasopharyngeal cancer; and blood cancers. , including: childhood leukemia, acute lymphocytic leukemia, acute myeloid leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, hairy cell leukemia, acute promyelocytic leukemia, plasma cell leukemia; bone marrow cancer blood diseases, including: myelodysplastic syndrome, myeloproliferative disorders, aplastic anemia, Fanconi anemia, essential macroglobulinemia; lung cancer, including: small cell lung cancer, non-small cell lung cancer; lymphoma, including: Hodgkin's disease , non-Hodgkin's lymphoma, cutaneous T-cell lymphoma, peripheral T-cell lymphoma, AIDS-related lymphoma; eye cancers, including retinoblastoma, uveal melanoma; skin cancers, including melanoma, non-melanoma skin cancer, Merkel cell carcinoma; soft tissue sarcomas, such as childhood soft tissue sarcoma, adult soft tissue sarcoma, Kaposi's sarcoma; urinary system cancers, including renal cancer, Wilms' tumor, bladder cancer, urethral cancer and metastatic cell carcinoma.

[0084] The compounds disclosed in the present invention can be used to treat cancers including breast cancer, lung cancer, liver cancer, ovarian cancer, prostate cancer, head and neck cancer, kidney cancer, stomach cancer and brain cancer.

[0085] Preferred cancers that can be treated by the compounds of the invention are solid tumors and hematological malignancies.

[0086] The term "unsubstituted" as used herein means having no substituents or being substituted only with hydrogen.

[0087] Some of the terms used in this invention are defined as follows:

[0088] "Halogen" refers to fluorine, chlorine, bromine and iodine.

[0089] "Isotope" refers to a compound that is identical to a compound listed herein, but in which one or more atoms are replaced by another atom having an atomic mass or mass number different from that normally found in nature. Isotopes that can be introduced into a compound of formula (I) include hydrogen, carbon, nitrogen, oxygen, fluorine, sulfur, i.e. 2 H, 3 H. 13 C. 14 C. 15 N. 17 O. 18 O. 18 F. 35 The compounds of formula (I) and their stereoisomers containing the aforementioned isotopes and / or other atomic isotopes, as well as pharmaceutically acceptable salts of the compounds and stereoisomers, are all within the scope of the present invention.

[0090] "=O" refers to an oxo group.

[0091] "-CF3" refers to trifluoromethyl.

[0092] "-C(O)-NH-" is "-amide-".

[0093] "-NH-C(O)-" is "-aminoacyl-".

[0094] "Carbonyl" means

[0095] "Alkyl," when used as a group or part of a group, refers to a straight-chain or branched aliphatic hydrocarbon group. Preferred alkyl groups are C1-C14 alkyl groups; more preferred are C1-C10 alkyl groups; and most preferred are C1-C6 alkyl groups, unless otherwise indicated. Examples of straight-chain or branched C1-C6 alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, 2-propyl, n-butyl, isobutyl, tert-butyl, hexyl, and the like.

[0096] "Bicyclic" and "fused ring" refer to cyclic groups formed by two or more ring structures sharing two adjacent carbon atoms. These include 4-14 membered saturated fused ring cycloalkyls and 6-14 membered partially saturated fused ring cycloalkyls, specific examples of which include but are not limited to the following structures:

[0097] "Spirocycle" refers to a structure containing 7-14 carbon atoms and / or heteroatoms, wherein at least two rings share an atom. The heteroatoms include nitrogen, oxygen, and sulfur. 7-14 membered spirocycles include 7-14 membered saturated spirocycles and 7-14 membered partially saturated spirocycles. 7-14 membered saturated spirocycles refer to cyclic groups in which all rings are saturated. Specific examples include, but are not limited to:

[0098] "Cycloalkyl" refers to a saturated or partially saturated monocyclic, bicyclic, fused, or spirocyclic carbocyclic ring. Rings consisting of 3-12 carbon atoms are preferred. Examples include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl. The definitions of "bicyclic," "fused," or "spirocyclic" are provided in the relevant sections herein.

[0099] "Heteroalkyl" refers to a straight-chain or branched alkyl group containing at least one heteroatom selected from S, O, and N in the main chain. Preferably, the heteroalkyl group contains 2-14 atoms. Heteroalkyl groups include, but are not limited to, ethers, thioethers, alkyl esters, secondary or tertiary alkylamines, and alkylsulfinic acids.

[0100] "Heterocycloalkyl" refers to a group in which one or more (preferably 1, 2, or 3) carbon atoms in the "cycloalkyl" group defined above are replaced by oxygen, nitrogen, phosphorus, boron, selenium, silicon, or sulfur atoms (preferably oxygen, sulfur, or nitrogen). The heterocycloalkyl and alkyl moieties are as defined herein. Preferably, the group contains 1-3 heteroatoms. Preferred rings are 3-14 membered rings (i.e., 3-14 membered heterocycloalkyls), and more preferably, 4-7 membered rings (i.e., 4-7 membered heterocycloalkyls). Heterocycloalkyls include, but are not limited to, pyrrolidinyl, dihydropyrrolyl, tetrahydropyrrolyl, dihydropyrazolyl, piperidinyl, morpholinyl, tetrahydrofuranyl, tetrahydrothiofuranyl, tetrahydropyranyl, oxirane, azirane, or 2-pyrazolinyl, as well as lactams, lactones, cyclic imides, and cyclic anhydrides. Heterocycloalkyls may be substituted with one or more substituents.

[0101] "Heterocycloalkylalkyl" refers to a (heterocycloalkyl-alkyl)- radical, where the heterocycloalkyl and alkyl moieties are as defined herein. Heterocycloalkylalkyl radicals include, but are not limited to, (2-tetrahydrofuranyl)methyl, (2-tetrahydrothiofuranyl)methyl, and the like.

[0102] "Alkylamino" includes both monoalkylamino and dialkylamino groups, unless otherwise specified. "Monoalkylamino" refers to a group consisting of (alkyl-NH)-; "dialkylamino" refers to a group consisting of ((alkyl)2N)-. The term "alkyl" is defined herein. The alkyl group is preferably a C1-C6 alkyl group. Examples include, but are not limited to, N-methylamino, N-ethylamino, N-isopropylamino, and N,N-(diethyl)amino.

[0103] "Heteroalkylamino" refers to both mono-heteroalkylamino and di-heteroalkylamino groups, unless otherwise indicated. Mono-heteroalkylamino refers to a group consisting of (heteroalkyl-)NH-; di-heteroalkylamino refers to a group consisting of (heteroalkyl)2N-. The "heteroalkyl" group is defined as such in the relevant text.

[0104] "Aminoalkyl" refers to a group consisting of (amino-alkyl)-. The "alkyl" portion is as defined herein. The aminoalkyl group is preferably an amino C1-C6 alkyl group. It should be noted that "amino-C1-C6 alkyl" refers to a C1-C6 alkyl group substituted with an amino group, examples of which include, but are not limited to, aminoethyl, 1-aminopropyl, and 2-aminopropyl.

[0105] "Arylamino" includes both mono-arylamino and di-arylamino, unless otherwise specified. Mono-arylamino refers to a group of formula (aryl-)NH-; di-arylamino refers to a group of formula (aryl)2N-; the definition of aryl is given in the relevant part of this document.

[0106] "Acyl" includes (alkyl-CO)- and (aryl-CO)- groups, unless otherwise indicated. Alkyl and aryl are as defined herein. Examples of acyl include, but are not limited to, acetyl, propionyl, isobutyryl, and benzoyl.

[0107] "Acylamide" includes (alkyl-C(O)NH)- and (aryl-C(O)NH)- groups, unless otherwise specified. Alkyl and aryl are as defined herein. Examples of amide include, but are not limited to, acetamido, propionamido, butyramido, isobutyramido, and benzamido.

[0108] "Alkenyl," as a group or part of a group, refers to an aliphatic hydrocarbon group containing at least one carbon-carbon double bond, which may be straight or branched. Alkenyl groups having C2-C14 are preferred. C2-C12 is more preferred; C2-C6 is most preferred. The group may contain multiple double bonds in its backbone, each in an E or Z configuration. Examples of alkenyl groups include, but are not limited to, ethenyl and propenyl.

[0109] "Alkynyl" as a group or part of a group refers to an aliphatic hydrocarbon group containing at least one carbon-carbon triple bond, which can be straight or branched. Preferably, it is C2-C14 alkynyl, more preferably C2-C12 alkynyl, and most preferably C2-C6 alkynyl. Examples of such alkynyl groups include, but are not limited to, ethynyl, prop-1-yn-1-yl, prop-2-yn-1-yl, but-1-yn-1-yl, but-3-yn-1-yl, 1-methylprop-2-yn-1-yl, pent-1-yn-1-yl, pent-4-yn-1-yl, hex-1-yn-1-yl, hex-5-yn-1-yl, and the like.

[0110] "Alkoxy" refers to a group of (alkyl-O)-. The "alkyl" part thereof is defined in the relevant text. The alkoxy group is preferably a C1-C8 alkoxy group, more preferably a C1-C6 alkoxy group. Examples of the alkoxy group include, but are not limited to: methoxy, ethoxy, n-propoxy, 1-methylethoxy, n-butoxy, isobutoxy, sec-butoxy, tert-butoxy, n-pentoxy, isopentoxy, neopentoxy, 1-methylbutoxy, 1-ethylpropoxy, n-hexyloxy, isohexyloxy, 3-methylpentoxy, 2-methylpentoxy, 1-methylpentoxy, 3,3-dimethylbutoxy, 2,2-dimethylbutoxy, 1,1-dimethylbutoxy, 1,2-dimethylbutoxy, 1,3-dimethylbutoxy, 2,3-dimethylbutoxy, 1-ethylbutoxy, 2-ethylbutoxy, etc. In addition, "alkoxycarbonyl" means a group in which the "alkoxy" defined above is bonded to a carbonyl group, for example, methoxycarbonyl, ethoxycarbonyl, etc.

[0111] "Alkenyloxy" refers to a group of (alkenyl-O)-, wherein alkenyl is as defined herein. C1-C6 alkenyloxy is preferred.

[0112] "Alkynyloxy" refers to a group of (alkynyl-O)-, wherein alkynyl is as defined herein. C1-C6 alkynyloxy is preferred.

[0113] "Alkoxycarbonyl" refers to a group consisting of (alkyl-OC(O))-. Alkyl is defined herein. Preferred alkyl groups are C1-C6 alkyl groups. Examples include, but are not limited to, methoxycarbonyl and ethoxycarbonyl.

[0114] "Alkylsulfinyl" refers to a group of (alkyl-S(O))-. "Alkyl" is defined herein. Preferred alkyl groups are C1-C6 alkyl groups. Alkylsulfinyl groups include, but are not limited to, methylsulfinyl, ethylsulfinyl, and the like.

[0115] "Alkylsulfonyl" refers to a group of (alkyl-S(O)2-O)-. "Alkyl" is defined herein. Preferred alkyl groups are C1-C6 alkyl groups. Examples include, but are not limited to, methylsulfonyl and ethylsulfonyl.

[0116] "Alkylaminocarbonyl" refers to an alkylamino-carbonyl group, wherein alkylamino is as defined herein.

[0117] "Cycloalkylalkyl" refers to a cycloalkyl-alkyl group. The cycloalkyl and alkyl moieties are as defined herein. Monocyclic alkyl groups include, but are not limited to, cyclopropylmethyl, cyclopentylmethyl, cyclohexylmethyl, and cycloheptylmethyl.

[0118] "Heterocycloalkenyl" refers to a heterocycloalkyl group containing at least one double bond. Heterocycloalkyl is defined herein.

[0119] "Aryl" as a group or part of a group refers to: (1) an aromatic monocyclic or fused ring; preferably an aromatic carbocyclic ring (a cyclic structure in which all the ring atoms are carbon) having 5-12 carbon atoms. Examples of aryl groups include, but are not limited to, phenyl and naphthyl; (2) partially saturated carbocyclic rings, such as phenyl and a C5-C7 cycloalkyl or C5-C7 cycloalkenyl group fused together to form a cyclic structure. Examples include, but are not limited to, tetrahydronaphthyl, indenyl, or hydroindenyl. Aryl groups may be substituted with one or more substituents.

[0120] "Arylalkenyl" refers to a group consisting of (aryl-alkenyl)-, wherein aryl and alkenyl are as defined herein. Exemplary arylalkenyl groups include, but are not limited to, phenylpropenyl and the like.

[0121] "Aralkyl" refers to a group consisting of (aryl-alkyl)-, wherein the aryl and alkyl portions are as defined herein. Exemplary aralkyl groups include, but are not limited to, benzyl, phenethyl, 1-naphthylmethyl, and the like.

[0122] "Cycloalkenyl" refers to a non-aromatic monocyclic or polycyclic ring system. It contains at least one carbon-carbon double bond and preferably has 5-10 carbon atoms per ring. Exemplary monocyclic cycloalkenyl rings include, but are not limited to, cyclopentene, cyclohexene, or cycloheptene. A cycloalkenyl group may be substituted with one or more substituents.

[0123] "Heteroaryl" refers to a monocyclic or condensed polycyclic aromatic heterocyclic group, preferably an aromatic group having one or more (preferably 3 to 14, more preferably 5 to 10, particularly preferably 5 or 6) carbon atoms, and one or more (preferably 1, 2, 3 or 4) oxygen, nitrogen, phosphorus or sulfur ring atoms (preferably O, S or N) as ring atoms, preferably a 4-15 membered heteroaryl group, more preferably a 5-7 membered heteroaryl group. Examples of the heteroaryl group include furyl, thienyl, pyrrolyl, pyrazolyl, triazolyl, thiazolyl, pyridinyl, pyrimidinyl, pyrazinyl, indolyl, benzimidazolyl, pyridinyl, imidazolyl, 3-phenylpyrrolyl, thiazolyl-oxazolyl, tetrazolyl, isoxazolyl, indazolyl, pyridazinyl, quinolyl, purinyl, carbazolyl, acridinyl, pyrimidinyl, 2,3'-bifuranyl and isoquinolyl.

[0124] "Heteroarylalkyl" refers to a group consisting of (heteroaryl-alkyl)-, wherein the heteroaryl and alkyl portions are as defined herein. Exemplary heteroarylalkyl groups include, but are not limited to, 2-furylmethyl, 3-furylmethyl, 2-pyridylmethyl, and the like.

[0125] "Alkoxy" refers to a group of (alkyl)-O-, wherein the "alkyl" portion is as defined herein.

[0126] "Cycloalkoxy" refers to a group of: (cycloalkyl)-O-, wherein the "cycloalkyl" portion is as defined herein.

[0127] "Heteroalkoxy" refers to a group of: (heteroalkyl)-O-, wherein the "heteroalkyl" portion is as defined herein.

[0128] "Aryloxy" refers to a group of (aryl)-O-, wherein the "aryl" part is as defined herein.

[0129] "Arylalkoxy" refers to a group of: (aryl-alkyl)-O-, wherein the "aryl" and "alkyl" parts are as defined herein.

[0130] "Heteroaryloxy" refers to a group of (heteroaryl)-O-, wherein the "heteroaryl" part is as defined herein.

[0131] "Heteroarylalkoxy" refers to a group of: (heteroaryl-alkyl)-O-, wherein the "heteroaryl" and "alkyl" parts are as defined herein.

[0132] "Heterocycloalkoxy" refers to a group of (heterocycloalkyl)-O-, wherein the "heterocycloalkyl" part is as defined herein.

[0133] "Heterocycloalkylamino" refers to both mono-heterocycloalkylamino and di-heterocycloalkylamino groups, unless otherwise indicated. Mono-heterocycloalkylamino refers to a group consisting of (heterocycloalkyl)-NH-; di-heterocycloalkylamino refers to a group consisting of (heterocycloalkyl)2-N-. The "heterocycloalkyl" group is defined herein.

[0134] "Arylalkylamino" refers to both mono-arylalkylamino and di-arylalkylamino, unless otherwise specified. Mono-arylalkylamino refers to a group consisting of (aryl-alkyl)-NH-; di-arylalkylamino refers to a group consisting of (aryl-alkyl)2-N-. The "aryl" and "alkyl" moieties are as defined herein.

[0135] "Cycloalkylamino" refers to both mono-cycloalkylamino and di-cycloalkylamino groups, unless otherwise specified. Mono-cycloalkylamino refers to a (cycloalkyl)-NH- group; di-arylalkylamino refers to a (cycloalkyl)2-N- group. The "cycloalkyl" portion is defined as such in the relevant text.

[0136] "Arylamino" refers to both mono-arylamino and di-arylamino groups, unless otherwise specified. Mono-arylamino refers to a group consisting of (aryl)-NH-; di-arylamino refers to a group consisting of (aryl)2-N-. The definition of "aryl" is given in the relevant text.

[0137] "Heteroarylamino" refers to both mono-heteroarylamino and di-heteroarylamino, unless otherwise indicated. Mono-heteroarylamino refers to a (heteroaryl)-NH- group; di-heteroarylamino refers to a (heteroaryl)2-N- group. The "heteroaryl" group is defined in the relevant sections herein.

[0138] "Heteroarylalkylamino" refers to both mono-heteroarylalkylamino and di-heteroarylalkylamino, unless otherwise indicated. Mono-heteroarylalkylamino refers to a (heteroaryl-alkyl)-NH- group; di-heteroarylalkylamino refers to a (heteroaryl-alkyl)2-N- group. The "heteroaryl" and "alkyl" moieties are as defined herein.

[0139] Unless otherwise specified, the subunit of the present invention refers to a divalent group, that is, a group in which one hydrogen atom in a monovalent group is replaced by a valency. For example, "heteroalkylene" refers to a heteroalkyl group in which one hydrogen atom is replaced by a valence; "heterocyclylene" refers to a heterocyclyl group in which one hydrogen atom is replaced by a valence; "arylene" refers to an aryl group in which one hydrogen atom is replaced by a valence; "alkylene" refers to an alkyl group in which one hydrogen atom is replaced by a valence; "alkenylene" refers to an alkenyl group in which one hydrogen atom is replaced by a valence; "cycloalkylene" refers to a cycloalkyl group in which one hydrogen atom is replaced by a valence; "heteroarylene" refers to a heteroaryl group in which one hydrogen atom is replaced by a valence; "heterocycloalkylene" refers to a heterocycloalkyl group in which one hydrogen atom is replaced by a valence; "heterocycloalkenylene" refers to a heterocycloalkenyl group in which one hydrogen atom is replaced by a valence; "alkyleneoxy" refers to an alkoxy group in which one hydrogen atom is replaced by a valence; "alkenyleneoxy" refers to an alkenyloxy group in which one hydrogen atom is replaced by a valence; "alkynyleneoxy" refers to an alkynyloxy group in which one hydrogen atom is replaced by a valence, and the like. Wherein, the definitions of the above-mentioned heterocyclic group, aryl, alkyl, alkenyl, cycloalkyl, heteroaryl, heterocycloalkyl, heterocycloalkenyl, alkoxy, alkenyloxy, alkynyloxy, etc. can be found in the relevant definitions herein.

[0140] The present invention includes compounds represented by general formula (I) and their various possible isomeric forms, including diastereoisomers, mirror image isomers, tautomers, and geometric isomers of "E" or "Z" configuration isomers. Any chemist with a certain level of basic knowledge can isolate the above optically pure or stereoisomerically pure compounds.

[0141] The present invention includes the general formula (I) and the compounds represented by the general formula and possible racemates and / or mirror isomers and / or mixtures of diastereomers.

[0142] Furthermore, the compounds represented by general formula (I) also encompass both solvated and unsolvated forms of the compounds. Thus, each formula encompasses compounds having the indicated structures, including both hydrated and unhydrated forms thereof.

[0143] In addition to the compounds represented by general formula (I), kinase inhibitors according to various embodiments include pharmaceutically acceptable salts, prodrugs, and active metabolites of such compounds, and pharmaceutically acceptable salts of such metabolites.

[0144] The term "pharmaceutically acceptable salt" refers to certain salts of the above-mentioned compounds that can maintain their original biological activity and are suitable for medical use. Pharmaceutically acceptable salts of the compounds represented by general formula (I) are formed in two forms: one is a salt formed with an acid; the other is a salt formed with an alkali or an alkali metal. Acids that form pharmaceutically acceptable salts with the compounds represented by general formula (I) include inorganic acids and organic acids. Suitable inorganic acids include hydrochloric acid, sulfuric acid and phosphoric acid. Suitable organic acids can be selected from aliphatic, cycloaliphatic, aromatic, heterocyclic carboxylic acids and sulfonic acids; examples include but are not limited to formic acid, acetic acid, propionic acid, succinic acid, glycolic acid, gluconic acid, lactic acid, malic acid, tartaric acid, glycine, arginine, citric acid, fumaric acid, alkylsulfonic acid, aromatic sulfonic acid, etc. Alkali metals that form pharmaceutically acceptable salts with the compound represented by general formula (I) include lithium, sodium, potassium, magnesium, calcium, aluminum, zinc, etc.; bases that form pharmaceutically acceptable salts with the compound represented by general formula (I) include choline, diethanolamine, morpholine, etc.

[0145] A "prodrug" is a derivative represented by general formula (I) that is converted in vivo (e.g., by hydrolysis, reduction, or oxidation) into a compound represented by general formula (I) through in vivo metabolism. For example, a compound represented by general formula (I) containing a hydroxyl group can be reacted with an acid to form a corresponding ester. The corresponding ester is a prodrug and can then be hydrolyzed in vivo to form the parent drug. Acids suitable for preparing "prodrugs" include, but are not limited to, acetic acid, citric acid, lactic acid, tartaric acid, malonic acid, oxalic acid, salicylic acid, succinic acid, fumaric acid, maleic acid, methylene-bis-β-hydroxynaphthoic acid, gentisic acid, isethionic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, and the like.

[0146] The kinase inhibitors referred to in the present invention include IC 50 The kinases used in the present invention include, but are not limited to, RET, PDGFR, VEGFR, and B-RAF.

[0147] The compound represented by general formula (I) can be administered either enterally or parenterally. Enterally: Orally or rectally. Parenteral: Subcutaneously, intramuscularly, intravenously, and intradermally. Generally, the active compound represented by general formula (I) can be administered with a pharmaceutically acceptable carrier or diluent.

[0148] "Therapeutically effective amount" or "therapeutic amount" refers to an amount sufficient to produce a therapeutic effect. An effective amount can be administered in one or more divided doses. Generally, an effective amount is sufficient to alleviate, improve, stabilize, slow, or delay further progression of a disease.

[0149] The compounds of the present invention may be used alone or in combination with one or more other drugs; or in combination with surgery or radiotherapy; or formulated with pharmaceutically acceptable carriers, diluents, or excipients to form a specific dosage form for administration. The specific dosage form depends on the route of administration.

[0150] The parenteral injection drug formulation of the present invention comprises a pharmaceutically acceptable sterile aqueous solution or non-aqueous solution, a dispersant, a suspending agent or an emulsifier, and a powder injection which is prepared into an injectable sterile aqueous solution before use.

[0151] If desired, and for more effective distribution, the compounds of the invention can be incorporated into slow-release or targeted-delivery systems such as polymer matrices, liposomes, and microspheres.

[0152] Solid dosage forms for oral administration include capsules, tablets, troches, powders and granules. In these solid dosage forms, the active compound represented by general formula (I) is mixed with at least one inert and pharmaceutically acceptable excipient or carrier. These excipients or carriers include sodium citrate or dicalcium phosphate and / or a) fillers or extenders, such as starch, lactose, sucrose, glucose, mannitol and salicylic acid; b) binders, such as carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose and acacia; c) disintegrants, such as agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates and sodium carbonate; d) dissolution delaying agents, such as paraffin; e) absorption accelerators, such as quaternary ammonium compounds; f) wetting agents, such as cetyl alcohol and glyceryl monostearate; g) adsorbents, such as kaolin and bentonite; and h) lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols.

[0153] Solid dosage forms of tablets, dragees, capsules, troches, and granules can be prepared with coatings or shells.

[0154] The active compound can also be administered in microencapsulated form. If desired, one or more of the above-mentioned excipients can be present.

[0155] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, etc. In addition to the active compound, the liquid dosage form may contain inert diluents commonly used in the art, such as water or other solvents, stabilizers and emulsifiers, such as ethyl alcohol, ethyl carbonate, ethyl acetate, benzoic acid alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils (especially cottonseed, peanut, corn, germ, olive, castor and sesame oils), glycerol, tetrahydrofuranol, polyethylene glycol and fatty acid esters of sorbitan, etc.

[0156] Besides inert diluents, the oral compositions can also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

[0157] The suspension may contain, in addition to the active compound, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan esters.

[0158] Compositions for rectal or vaginal administration are preferably in the form of suppositories. These can be prepared by mixing the compounds of the invention with suitable non-irritating excipients or carriers.

[0159] Dosage forms for topical administration of the compounds of this invention include powders, patches, sprays, ointments and inhalants. The active compound is mixed under sterile conditions with a pharmaceutically acceptable carrier and any desired preservatives, buffers or propellants.

[0160] The preferred dosage range is about 0.01-400 mg per kilogram of body weight per day. A more preferred dosage range is 0.2-100 mg per kilogram of body weight per day. Alternatively, the appropriate dosage may be administered in divided doses over a period of several days.

[0161] The compounds of the present invention can be used as, but not limited to, kinase inhibitors. The pyrimidine derivatives disclosed herein can be used alone or in combination with other drugs or pharmaceutically acceptable carriers, diluents, or excipients, and are suitable for preventing or treating conditions caused by, associated with, or accompanied by disruption of cell proliferation and / or angiogenesis. One example of such conditions is cancer.

[0162] 1. The compounds of the present invention can also be used to treat diseases that involve or are at least partially regulated by the activity of PDGFR, VEGFR, FGFR, FLT3, Aurora-A, Aurora-B, TRK, RAF, RET or Abl, wherein RET, PDGFR, VEGFR activity is known to play a role in promoting the onset of the disease, or such symptoms are known or have been shown to be alleviated by RET, PDGFR, VEGFR inhibitors. Conditions of this type that are expected to be treatable by the compounds of the present invention include, but are not limited to, the following: antiproliferative conditions (e.g., cancer); neurodegenerative diseases including Huntington's disease, polyglutamine diseases, Parkinson's disease, Alzheimer's disease, epileptic seizures, striatonigral degeneration, progressive supranuclear palsy, torsion dystonia, spasmodic torticollis and movement disorders, familial tremor, Tourette syndrome, diffuse Lewy body disease, progressive supranuclear palsy, Pick's disease, intracranial hemorrhage, primary lateral sclerosis, spinal muscular atrophy, amyotrophic lateral sclerosis, hypertrophic interstitial polyneuropathy, retinitis pigmentosa, hereditary optic atrophy, hereditary spastic paraplegia, progressive ataxia, and Shy-Drager syndrome; metabolic diseases including type 2 diabetes; degenerative eye diseases including glaucoma, age-related macular degeneration, rubroiridis glaucoma; inflammatory diseases and / or immune system disorders including rheumatoid arthritis (RA), osteoarthritis, Arthritis, juvenile chronic arthritis, graft-versus-host disease, psoriasis, asthma, spondyloarthropathies, psoriasis, Crohn's disease, inflammatory bowel disease, colon ulcers, alcoholic hepatitis, diabetes, Sjoegren's syndrome, multiple sclerosis, ankylosing spondylitis, membranous glomerulopathy, disc pain, systemic lupus erythematosus; diseases involving angiogenesis, including cancer, psoriasis, and rheumatoid arthritis; psychological disorders, including bipolar disorder and schizophrenia , mania, depression and dementia; cardiovascular diseases including heart failure, restenosis and arteriosclerosis; fibrotic diseases including liver fibrosis, cystic fibrosis and angiofibroma; infectious diseases including fungal infections such as Candida albicans and bacterial infections; viral infections such as herpes simplex; protozoal infections such as malaria, Leishmania infection, Trypanosoma brucei infection, toxoplasmosis and coccidiosis; hematopoietic disorders including thalassemia, anemia and sickle cell anemia.

[0163] The term "cancer" as used herein generally refers to a broad range of diseases characterized by uncontrolled abnormal growth of cells.

[0164] The compounds of the present invention are expected to be useful in treating various cancers, including but not limited to: bone cancers, including Ewing's sarcoma, osteosarcoma, chondrosarcoma, etc.; brain and CNS tumors, including acoustic neuroma, neuroblastoma, glioma and other brain tumors, spinal cord tumors, breast cancer, colorectal cancer, advanced colorectal adenocarcinoma; endocrine cancers, including adrenocortical carcinoma, pancreatic cancer, pituitary cancer, thyroid cancer, parathyroid cancer, thymic carcinoma, multiple endocrine neoplasia; gastrointestinal cancers, These include: gastric cancer, esophageal cancer, small intestinal cancer, liver cancer, extrahepatic bile duct cancer, gastrointestinal carcinoid tumors, and gallbladder cancer; genitourinary cancers, including: testicular cancer, penile cancer, and prostate cancer; gynecological cancers, including: cervical cancer, ovarian cancer, vaginal cancer, uterine / endometrial cancer, pudendal cancer, gestational trophoblastic tumor, fallopian tube cancer, and uterine sarcoma; head and neck cancers, including: oral cancer, lip cancer, salivary gland cancer, laryngeal cancer, hypopharyngeal cancer, pharyngeal cancer, nasal cancer, paranasal sinus cancer, and nasopharyngeal cancer; and blood cancers. , including: childhood leukemia, acute lymphocytic leukemia, acute myeloid leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, hairy cell leukemia, acute promyelocytic leukemia, plasma cell leukemia; bone marrow cancer blood diseases, including: myelodysplastic syndrome, myeloproliferative disorders, aplastic anemia, Fanconi anemia, essential macroglobulinemia; lung cancer, including: small cell lung cancer, non-small cell lung cancer; lymphoma, including: Hodgkin's disease , non-Hodgkin's lymphoma, cutaneous T-cell lymphoma, peripheral T-cell lymphoma, AIDS-related lymphoma; eye cancers, including retinoblastoma, uveal melanoma; skin cancers, including melanoma, non-melanoma skin cancer, Merkel cell carcinoma; soft tissue sarcomas, such as childhood soft tissue sarcoma, adult soft tissue sarcoma, Kaposi's sarcoma; urinary system cancers, including renal cancer, Wilms' tumor, bladder cancer, urethral cancer and metastatic cell carcinoma.

[0165] Cancers that the compounds of the present invention can be used to treat include, but are not limited to, breast cancer, lung cancer, ovarian cancer, thyroid cancer, colorectal cancer, prostate cancer, head and neck cancer, kidney cancer, stomach cancer, and brain cancer.

[0166] Preferred cancers that can be treated by the compounds of the present invention are solid tumors and hematological malignancies.

[0167] In addition, the compounds of the present invention can be used to treat proliferative diseases that are resistant to other chemotherapy treatments; and for the treatment of hyperproliferative diseases, such as leukemia, psoriasis, etc.

[0168] Synthesis of 3-aminoindoline derivatives

[0169] The compounds represented by general formula (I) can be synthesized using the synthetic routes and methods discussed below. The raw materials used are readily available. However, the synthetic routes and methods used in the present invention can be widely applied to the synthesis of analogous compounds, requiring only the substitution of the starting materials. For example, the synthesis of compounds not described in detail in the examples herein can be achieved by replacing the starting materials with those of the corresponding target compound and, if necessary, slightly modifying the reaction conditions based on common chemical knowledge to produce the desired target compound.

[0170] The reagents of each embodiment can be prepared using the reaction pathways or synthetic flow charts described below. The preparation of specific compounds of the embodiments is described in detail in the following examples. However, those skilled in the art will appreciate that the chemical reactions described can be applied to the preparation of a variety of other compounds in different embodiments. For example, the synthesis of non-exemplified compounds can be successfully carried out by modifications obvious to those skilled in the art, or by changing to other appropriate reagents known in the art. A list of suitable protecting groups in organic synthesis can be found in TW Greene's Protective Groups in Organic Synthesis, John Wiley & Sons, 1981. Other reactions disclosed herein or known in the art can be considered to have applicability for preparing other compounds of each embodiment.

[0171] Reagents useful in synthesizing the compounds can be obtained or prepared according to techniques known in the art.

[0172] In the following examples, all temperatures are in degrees Celsius unless otherwise indicated.

[0173] All starting materials and reagents were commercially available, including but not limited to Aldrich Chemical Company, Lancaster Synthesis Ltd, etc. Commercially available starting materials and reagents were used without further purification unless otherwise indicated.

[0174] Glassware was oven-dried and / or heat-dried. Reactions were followed on glass silica gel-60F254 plates (0.25 mm) (TLC). Analytical thin-layer chromatography was performed using an appropriate solvent ratio (v / v). The reaction endpoint was determined by TLC when the starting material was consumed.

[0175] Typically, subsequent workup involves doubling the volume of the reaction solution with the solvent used in the reaction, followed by three extractions with 25% of the total volume of the extraction solvent, unless otherwise specified. The product-containing extract is dried over anhydrous sodium sulfate, filtered on a rotary evaporator, and the solvent is evaporated under reduced pressure, ensuring that the solvent is removed in vacuo. Finally, the target compound is isolated by flash column chromatography (J. Org. Chem., 1978; 43: 2923).

[0176] 1 H NMR spectra were obtained using a Bruker instrument (400 MHz), with chemical shifts expressed in ppm. Chloroform (7.25 ppm) or tetramethylsilane (0.00 ppm) was used as the reference standard. Other commonly used NMR solvents may also be used, if desired. 1 H NMR notation: s = singlet, d = doublet, t = triplet, m = multiplet, br = broadened, dd = doublet of a doublet, dt = doublet of a triplet. Coupling constants, when given, are given in Hz.

[0177] Mass spectra were obtained using LC / MS, using either ESI or APCI ionization. All melting points are uncorrected.

[0178] The following examples are intended solely to illustrate the synthesis of specific compounds of the invention. However, the synthetic methods are not limiting in any way. Compounds not listed below can also be prepared using the same synthetic routes and methods as those described below, selecting appropriate starting materials and, where necessary, adjusting the reaction conditions using common sense.

[0179] synthesis

[0180] The compound shown in the general formula (I), when (R 6 )N can be linked to the C of the indoline ring 5 - position, and R 1 =R 3 =R 4 =R 6 =H, R 2 =OCH3, and Y = carbon atom, the synthesis method is as follows: A suitably substituted 5-nitro-indoline derivative (VI) reacts with a suitably chlorinated compound (VII) to yield the corresponding 5-nitro-substituted indoline derivative (VIII). The latter then reacts with a compound (IX) containing an active hydrogen under heating in toluene to yield compound (X). Compound (X) is reduced to yield compound (XI), which is then reacted with 3-methoxymethylene-oxoindoline-6-carboxylate (XII) to yield the target compound represented by general formula (I), i.e., (XIII). See Synthesis Route 1 for details.

[0181] Synthesis Route 1

[0182] The compound shown in the general formula (I), when (R 6 )N can be linked to the C 5 - position, and R 1 =R 3 =R 5 =R 6 =H, R 2 =OCH3, X = -C(O)-, and Y = carbon atom, the synthesis method is as follows: A suitably substituted 4-bromo-2-oxoindoline-6-carboxylic acid methyl ester (XIV) reacts with a boron derivative (XV) in the presence of a palladium reagent to produce compound (XVI). Compound (XVI) reacts with triethyl orthoformate in a suitable acid as a solvent to produce compound (XVII). Separately, a suitable 5-nitroindoline derivative (XVIII) reacts with a suitable compound (XIX) in the presence of a base to produce compound (XX). The latter is reduced to produce the important intermediate (XXI). Compound (XVII) undergoes a substitution reaction with the intermediate to produce the target compound (XXII) represented by general formula (I). See Synthesis Route 2 for details.

[0183] Synthesis Route 2

[0184] The following examples further illustrate the content of the present invention. The purpose is to enable those skilled in the art to more clearly understand and practice the specific content of the present invention. However, the scope of protection of the present invention is not limited to these examples.

[0185] Synthesis of methyl (Z)-3-(((1-(2-((1R,4R)-2-oxa-5-azabicyclo[2.2.1]heptane-5-yl)ethyl)indolin-5-yl)amino)methylene)-4-cyclopropyl-2-oxoindolin-6-carboxylate (1)

[0186] Step-1 Synthesis of methyl 4-cyclopropyl-2-oxoindoline-6-carboxylate (XVI-1)

[0187] To a solution of compound (IV-1) (20.0 g, 0.0741 mol, 1.0 eq) in toluene (400.0 mL, 20.0 Vol) and water (66.6 mL, 3.3 Vol) were added cyclopropylboronic acid (XV-1, 12.7 g, 0.1481 mol, 2.0 eq), palladium acetate (1.0 g, 4.4460 mmol, 0.06 eq), tricyclohexylphosphine (2.1 g, 74.1000 mmol, 0.10 eq), and potassium phosphate (31.4 g, 0.1482 mol, 2.0 eq). Under nitrogen, the system was heated to 100°C and stirred for 11.5 hours. The reaction mixture was cooled to room temperature, and ethyl acetate (1.0 L) and water (400.0 mL) were added. The mixture was separated, and the organic phase was washed with water (300.0 mL x 3) and dried over anhydrous sodium sulfate. The mixture was filtered, and the filtrate was concentrated under reduced pressure to remove the solvent. The residue was passed through a silica gel column to obtain the title solid compound (XVI-1) (8.0 g, 46.7%). 1 H NMR (400MHz, DMSO-d6) δ10.53(s,1H),7.15(d,J=1.4Hz,1H),7.08(d,J=1.4Hz,1H),3.82(s,3H), 3.61(s,2H),1.98-1.82(m,1H),1.04-0.94(m,2H),0.79-0.69(m,2H).ESI-MS(m / z):232.2[M+H] + .

[0188] Step-2 Synthesis of methyl 4-cyclopropyl-3-(ethoxymethylene)-2-oxoindoline-6-carboxylate (XVII-1)

[0189] To a solution of compound (XVI-1) (19.5 g, 84.4541 mmol, 1.0 eq) in acetic acid (254.0 mL, 13.0 Vol) was added triethyl orthoformate (125.2 g, 844.5405 mmol, 10.0 eq). The system was heated to 100°C and allowed to react for 17 hours. The system was cooled to room temperature and concentrated under reduced pressure to remove the solvent to obtain a crude product. Ethyl acetate (220.0 mL), ethanol (45.0 mL), and petroleum ether (293.0 mL) were added to the crude product, stirred at 70°C for 2 hours, cooled to approximately 50°C, and filtered while hot. The filter cake was rinsed with 60.0 mL of a mixed solvent (ethyl acetate:petroleum ether = 1:1) and then 40.0 mL of petroleum ether. The mixture was then air-dried at 60°C for 17 hours to obtain the title compound (XVII-1) (15.8 g, 65.4%). 1H NMR (400MHz, DMSO-d6) δ10.50(s,1H),7.65(s,1H),7.16(d,J=1.5Hz,1H),7.01(d,J=1.5Hz,1H),4.40(q,J=7.1Hz,2H),3.81 (s,3H),2.79(tt,J=8.5,5.3Hz,1H),1.32(t,J=7.1Hz,3H),1.02-0.90(m,2H),0.76-0.65(m,2H).ESI-MS(m / z):288.2[M+H] + .

[0190] Step-3 Synthesis of 2-(5-nitroindol-1-yl)ethan-1-ol (XX-1)

[0191] 5-Nitroindoline (XVIII-1) (50.0 g, 304.5809 mmol, 1.0 eq) was added to N,N-dimethylformamide (500.0 mL, 10.0 Vol) and mechanically stirred until dissolved. Cesium carbonate (198.5 g, 609.1615 mmol, 2.0 eq), potassium iodide (5.1 g, 30.4581 mmol, 0.1 eq), and bromoethanol (XIX-1, 57.1 g, 456.8713 mmol, 1.5 eq) were added and heated to 100°C under argon for 16 hours. After cooling to room temperature, water (1.0 L) was added and the mixture was extracted with ethyl acetate (1.0 L x 1). The organic phase was separated, washed with saturated brine (1.0 L x 4), and dried over anhydrous sodium sulfate. The filtrate was filtered and concentrated under reduced pressure to afford the title compound XX-1 (70.0 g, 110.4%).

[0192] Step-4 Synthesis of (1-(2-((1R,4R)-2-oxa-5-azabicyclo[2.2.1]heptane-5-yl)ethyl)indole-5-amine (XX-1)

[0193] Compound (IX-1) (48.5 mg, 0.1856 mmol, 1.20 eq) was added to a methanol solution (1.5 mL, 30.0 Vol) of compound (X-1) (50.0 mg, 0.1546 mmol, 1.0 eq). The mixture was stirred and reacted at 70-75°C under nitrogen for 10-15 hours. The mixture was cooled to room temperature, filtered, and washed with methanol to obtain the title compound (XI-1) (XRI-0001-061-19, 65.0 mg, 78.0%). 1H NMR (400MHz, CDCl3) δ12.08(s,1H),8.27(br,1H),7.96-7.36(m,8H),6.69(s,1H),6.62(dd,J=8.8,2.3Hz,1H),5.96(d,J=8.28,1H),4 .12(t,J=8.4Hz,2H),3.85(s,3H),3.75(t,J=4.7Hz,4H),3.21(s,2H),3.04(t,J=8.4Hz,2H),2.58(br,4H).ESI-MS(m / z):539.2[M+H] + .

[0194] (A) Compound (XX-1) (70.0 g, 0.3362 mol, 1.0 eq) was added to dichloromethane (700.0 mL, 10.0 Vol) and stirred to dissolve. Triethylamine (68.0 g, 0.6724 mol, 2.0 eq) and p-toluenesulfonyl chloride (70.5 g, 0.3698 mol, 1.1 eq) were added under an ice bath and reacted at room temperature under nitrogen for 12 hours. Water (500.0 mL) was added and stirred for 3 hours. The organic phase was separated and saturated sodium bicarbonate solution (1.0 L) was added to the organic phase and stirred for 5 hours. The organic phase was separated and saturated sodium bicarbonate solution (1.0 L) was added to the organic phase and stirred overnight. The organic phase was separated and washed with saturated brine (1.0 L x 2). The organic phase was separated and dried over anhydrous sodium sulfate. Filtered and concentrated to obtain a crude brown solid. The solid was dispersed in ethyl acetate (200.0 mL) and stirred for 3 hours. The residue was filtered and dried to obtain the title compound ITM-1 (75.0 g, 61.6%).

[0195] (B) Compound (ITM-1) (200.0 mg, 0.5519 mmol, 1.0 eq) was dissolved in acetonitrile (5.0 mL, 25.0 Vol), followed by the addition of compound X-1 (89.8 mg, 0.6623 mmol, 1.2 eq), potassium carbonate (228.8 mg, 1.6557 mmol, 3.0 eq), and potassium iodide (18.3 mg, 0.1104 mmol, 0.2 eq). The mixture was reacted at 80°C overnight under nitrogen. After cooling to room temperature, water (20.0 mL) and ethyl acetate (20.0 mL) were added, and the organic phase was separated. The organic phase was washed with saturated brine (20.0 mL x 1), separated, and dried over anhydrous sodium sulfate. The residue was filtered, concentrated, and passed through a silica gel column to afford the title compound ITM-2 (110.0 mg, 68.9%).

[0196] (C) Compound (ITM-2) (110.0 mg, 0.3802 mmol, 1.0 eq) was dissolved in methanol (5.0 mL, 45.5 Vol). Pd / C (50.0 mg, 10% wt%) was then added and reacted at room temperature with hydrogen for 1 hour. After the reaction, the Pd / C was removed by filtration. The filtrate was concentrated to afford the title compound XX-1 (90.0 mg, 91.3%).

[0197] Step-5 Synthesis of (Z)-3-(((1-(2-((1R,4R)-2-oxa-5-azabicyclo[2.2.1]heptane-5-yl)ethyl)indolin-5-yl)amino)methylene)-4-cyclopropyl-2-oxoindolin-6-carboxylic acid methyl ester (XXII-1)

[0198] Compound (XVII-1) (89.7 mg, 0.3124 mmol, 0.9 eq) was added to a methanol solution (5.0 mL, 55.6 Vol) of compound (XXI-1) (90.0 mg, 0.3471 mmol, 1.0 eq). The mixture was reacted at 65°C overnight under nitrogen. The mixture was cooled to room temperature, filtered, and washed with methanol to obtain the title compound XXII-1 (120.0 mg, 76.5%). 1 H NMR(400MHz,DMSO-d6)δ11.44(d,J=12.9Hz 1H),10.73(s,1H),8.56(d,J=12.9Hz,1H),7.37(d,J=2.2Hz,1H),7.32(d,J=1.5Hz,1H),7.16(d,J=2.3Hz,1H),7.02(dd,J=8.4, 2.4Hz,1H),6.53(d,J=8.4Hz,1H),4.33(t,J=2.0Hz,1H),3.84(d,J=7.2Hz,1H),3.81(s,3H),3.54-3.48(m,2H),3.41(td,J=8.5 ,2.1Hz,2H),3.11(t,J=7.1Hz,2H),2.94(t,J=8.4Hz,2H),2.85(dd,J=9.9,1.8Hz,1H),2.79-2.66(m,2H),2.47-2.43(m,1H),2. 29-2.22(m,1H),1.72(dd,J=9.7,2.1Hz,1H),1.60-1.54(m,1H),1.09-1.01(m,2H),0.72-0.63(m,2H).ESI-MS(m / z):501.4[M+H] + .

[0199] Example 2-12

[0200] According to the method of Example 1, a wide variety of derivatives can be synthesized by simply changing the appropriate starting materials. Examples 2-12 are some representative examples (see Table 1).

[0201] Table 1 Representative 2-oxoindoline derivatives of general formula (I)

[0202] Biological experiments and pharmacodynamic analysis

[0203] 1. Detection of kinase activity

[0204] There are numerous reports on kinase activity testing in the literature, and relevant kinase assay kits are also available. Cisbio products include, but are not limited to, the HTRF kinase-STK KIT. Using the HTRF (homogeneous time-resolved fluorescence) kinase assay kit to detect RET kinase inhibitory activity as an example, the experimental steps are as follows.

[0205] 1. Experimental methods and steps:

[0206] 1.1 Prepare the test compound into a 10 mM (mmol / L) DMSO solution.

[0207] 1.2 The 10mM concentration of the compound was diluted with kinase buffer to 2.5μM (2.5× compound) working solution, and then 2.5μM was used as the highest concentration, and the 2.5× working solution of the test compound was continuously diluted to 9 concentrations in a 3-fold gradient, namely: 2.5, 0.833333, 0.277778, 0.092593, 0.030864, ​​0.010288, 0.003429, 0.001143, 0.000381μM; the 10mM control compound was diluted with kinase buffer. The buffer was diluted to a 2.5 μM working solution, and then 2.5 μM was used as the highest concentration. The 2.5× working solution of the test compound was continuously diluted to 9 concentrations in a 3-fold gradient dilution. The concentrations were: 2.5, 0.833333, 0.277778, 0.092593, 0.030864, ​​0.010288, 0.003429, 0.001143, and 0.000381 μM.

[0208] 1.3 Add 4 μL of 2.5× compound working solution to a 384-well plate (Greiner, Cat#781280). Set up blank wells (no compound or kinase) and control wells (only kinase without compound). Add 4 μL of kinase buffer to the blank and control wells.

[0209] 1.4 Prepare the kinase stock solution with kinase buffer to the corresponding 5× working solution. Add 2 μL of 5× kinase working solution to each well containing compound working solution, add 2 μL of kinase buffer to the Blank well, and add 2 μL of 5× kinase working solution to the Control well.

[0210] 1.5 Take 2 μL of 5× substrate stock solution (TK Antibody-Cryptate) and add it to each well containing compound and kinase working solution. Add 2 μL of kinase buffer to the blank well and 2 μL of 5× substrate stock solution to the control well.

[0211] 1.6 Take 2 μL of ATP working solution (5×) and add it to each detection well.

[0212] 1.7 Cover the 384-well plate with sealing film and incubate at 37°C for 1 hour. Then, add 5 μL (4X) reaction stop solution (Streptavidin-XL665) to each well.

[0213] 1.8 Continue to seal the 384-well plate with film, incubate at 37°C for 1 hour, and then measure the 665 and 620 signal values ​​on a 2104 EnVision plate reader.

[0214] Preparation of detection system working solution:

[0215] 2. Data Analysis:

[0216] The inhibition rate (IR) of the test compound was calculated using the following formula: IR (%) = (RLU CTR (665 / 620) – RLU compound (665 / 620)) / (RLU CTR (665 / 620) – RLU BLANK (665 / 620)) * 100%. The inhibition rate of the compound at different concentrations was calculated in Excel, and the IC was then calculated using GraphPad Prism 5 software. 50 .

[0217] 3. Activity inhibition test results of wild-type RET and mutant RET kinases

[0218] Some biological activity test results are shown in Table 2

[0219] Table 2 Inhibitory activity of target compounds on RET kinase

[0220] The target compounds claimed in the present invention have good inhibitory activity on RET kinase.

[0221] At the same time, the present invention also uses the compounds of the present invention to test other kinases, and it is also found that the compounds of the present invention also have a significant inhibitory effect on kinases (PDGFR (PDGFRα and PDGFRβ), VEGFR, FGFR, FLT3, Aurora-A, Aurora-B, TRK, B-RAF, RET or Abl activity), see Tables 3 and 4.

[0222] Table 3 Inhibitory activity of target compounds on VEGFR2 kinase

[0223] Table 4 Inhibitory activity of target compounds on PDGFRa kinase

[0224] Example 103

[0225] The cell activity test uses CTG (CELL TITER-GLO) luminescence method to test the activity of the target compound. Its principle is: ATP adenosine triphosphate (ATP) participates in a variety of enzymatic reactions in the body and is an indicator of living cell metabolism. Its content directly reflects the number and state of cells. During the experiment, an equal volume of CellTiter-Glo was added to the cell culture medium. TM Reagents measure luminescence values. In the light signal and system, the luminescence value is proportional to the amount of ATP, which is positively correlated with the number of living cells and inversely proportional to the activity of anti-tumor drugs. By detecting the fluorescence signal of ATP, the anti-proliferative activity data of the compound on tumor cells can be obtained according to the calculation formula.

[0226] The test of tumor cell inhibitory activity was carried out in two parts: using kinase as the target, the target compound was examined for its effects on human thyroid ductal carcinoma cells (TT) and human colon cancer cells (KM12); to determine its activity on RET Fusion, the target compound was specifically tested for its growth inhibitory activity on KIF5B-RET fusion cells.

[0227] Reagents used in the experiment: F-12K basal medium (ATCC, 30-2004), fetal bovine serum (Corning, 35-076-CV), double antibody (GIBCO, 15240-062), trypsin (GIBCO, 25200072), DMSO (SIGMA, D2650), DMEM basal medium (Corning, 10-013-CV), fetal bovine serum (Gibco, 10091-148).

[0228] 1. Activity assay of compounds against human thyroid ductal carcinoma cells and human colon cancer cells

[0229] The following method is used to determine the effect of compounds on tumor cell proliferation by using the CTG luminescence method.

[0230] Specific experimental operation methods and procedures:

[0231] 1.1 Cell recovery

[0232] Immediately remove the frozen cells from the liquid nitrogen storage tank and place them in a 37°C constant temperature water bath for 2 minutes. After the cell freezing solution is completely thawed, transfer the cell suspension into a 15 mL centrifuge tube, slowly add 4 mL of culture solution, centrifuge (1000 rpm, 5 minutes), discard the supernatant, aspirate the original solution, add 5 mL of the above culture medium, gently pipette to a single cell suspension, transfer it to a culture flask, and culture it in an incubator.

[0233] 1.2 Cell culture

[0234] The cells were cultured in complete medium in an incubator at 37°C with 5% CO2. The cells were passaged regularly and cells in the logarithmic growth phase were used for plating.

[0235] 1.3 Cell plating

[0236] Cells were stained with trypan blue and viable cells were counted. The cell concentration was adjusted to the appropriate medium plate concentration (TT: 50,000 cells / mL, KM12: 35,000 cells / mL). 90 μL of the cell suspension was added to each well of a 96-well culture plate (Corning, 3599). Blank and vehicle control wells were set up. Culture medium containing cells was added to the blank control wells, while culture medium without cells was added to the vehicle control wells. The culture plates were then placed in an incubator at 37°C, 5% CO2, and 100% relative humidity and incubated overnight.

[0237] 1.4 Compound preparation

[0238] Weigh the compound and prepare a 10 mM stock solution in DMSO. Dilute the stock solution of the test compound to a final concentration of 100 μM in serum-free medium on a dispensing plate (Beaver, Suzhou) to obtain a 10× working solution of the compound (including the control). Dilutions were performed in serum-free medium in a 3-fold concentration gradient to obtain nine 10× working solutions with compound concentrations of 100, 33.33, 11.11, 3.70, 1.23, 0.411, 0.137, 0.046, and 0.015 μM, respectively.

[0239] 1.5 Addition of compounds

[0240] Add 10x compound working solution at varying concentrations to a 96-well cell culture plate at 10 μL / well. Add 10 μL of DMSO-cell culture medium mixture to vehicle control and blank control wells for a final DMSO concentration of 0.1%. Set up two replicates for each concentration. Return the 96-well plate to a 37°C, 5% CO2 incubator and culture for 5 days.

[0241] 1.6CTG detection

[0242] The cell culture plate was removed and allowed to equilibrate to room temperature for 30 minutes. 50 μL (equal to half the volume of the cell culture medium in each well) of CellTiter-Glo working solution was added to each well. The cell plate was wrapped with aluminum foil to avoid light. The culture plate was shaken on an orbital shaker for 2 minutes to induce cell lysis. The culture plate was placed at room temperature for 10 minutes to stabilize the luminescent signal, and the luminescent signal was detected on a 2104 EnVision plate reader.

[0243] 1.7 Data Analysis

[0244] The inhibition rate (IR) of the test compound was calculated using the following formula: IR (%) = (1 – (RLU compound – RLU blank control) / (RLU vehicle control – RLU blank control)) * 100%. Finally, nonlinear regression analysis was performed using the logarithm of compound concentration-inhibition rate in Graphpad Prism 5 software to obtain the IC value of the compound for inhibiting cell proliferation. 50 value.

[0245] Results The test results are shown in Table 5.

[0246] Table 5 Inhibitory activity of compounds on TT cells, KM12 cells and LC-2 / ad cells

[0247] The compound obtained in the present invention also has significant inhibitory activity on the growth of NU-16 cells, NCI-H1703 cells, and GIST-T1 cells.

[0248] Table 6 Growth inhibitory activity of NU-16 cells, NCI-H1703 cells, and GIST-T1 cells

[0249] The target compound claimed in the present invention has good inhibitory activity on human thyroid duct cancer cells and human colon cancer cells.

[0250] 2. Determination of compound activity against RET fusion engineered cell lines

[0251] In addition to using relevant tumor cell lines, KIF5B-RET and CCDC6-RET fusion engineered cell lines were also used for in vitro anti-tumor inhibitory activity to specifically detect the activity of target compounds against KIF5B-RET Fusion and CCDC6-RET Fusion.

[0252] The following method was used to determine the effects of compounds on the proliferation of KIF5B-RET and CCDC6-RET fusion engineered cells using the CTG luminescence assay. For KIF5B-RET and CCDC6-RET fusions, Ba / F3-KIF5B-RET and Ba / F3-CCDC6-RET fusion engineered cell lines (created by Precedo) were cultured under the appropriate conditions.

[0253] Reagents used in the experiment: RPMI-1640 basal medium (GIBCO, 22400-089), fetal bovine serum (SH30084.03, SH30084.03), double antibody (GIBCO, 15240-062), trypsin (GIBCO, 25200072), DMSO (SIGMA, D2650).

[0254] The experimental operation method is as follows

[0255] 2.1 Cell recovery

[0256] Immediately remove the frozen cells from the liquid nitrogen storage tank and place them in a 37°C constant temperature water bath for 2 minutes. After the cell freezing solution is completely thawed, transfer the cell suspension into a 15 mL centrifuge tube, slowly add 4 mL of culture solution, centrifuge (1000 rpm, 5 minutes), discard the supernatant, aspirate the original solution, add 5 mL of the above culture medium, gently pipette to a single cell suspension, transfer it to a culture flask, and culture it in an incubator.

[0257] 2.2 Cell culture

[0258] Ba / F3-KIF5B-RET and Ba / F3-CCDC6-RET engineered cells were cultured in complete medium (RPMI-1640 + 10% FBS + 1% P / S) in a 37°C, 5% CO2 incubator. Cells were passaged regularly, and cells in the logarithmic growth phase were used for plating.

[0259] 2.3 Cell plating

[0260] Cells were stained with trypan blue and viable cells were counted. The cell concentration was adjusted to 30,000 cells / mL. 90 μL of the cell suspension was added to each well of a 96-well culture plate (Corning, 3599). Blank and vehicle control wells were set up. Culture medium containing cells was added to the blank control wells, while culture medium without cells was added to the vehicle control wells. The culture plates were then placed in an incubator at 37°C, 5% CO2, and 100% relative humidity and incubated overnight.

[0261] 2.4 Compound preparation

[0262] Weigh the compound and prepare a 10 mM stock solution in DMSO. Dilute the stock solution of the test compound to a final concentration of 100 μM in serum-free medium on a dispensing plate (Beaver, Suzhou) to obtain a 10× working solution of the compound (including the control). Dilutions were performed in serum-free medium in a 3-fold concentration gradient to obtain nine 10× working solutions with compound concentrations of 100, 33.33, 11.11, 3.70, 1.23, 0.411, 0.137, 0.046, and 0.015 μM, respectively.

[0263] 2.5 Addition of compounds

[0264] Add 10x compound working solution at varying concentrations to a 96-well cell culture plate at 10 μL / well. Add 10 μL of DMSO-cell culture medium mixture to vehicle control and blank control wells for a final DMSO concentration of 0.1%. Set up two replicates for each concentration. Return the 96-well plate to a 37°C, 5% CO2 incubator and culture for 5 days.

[0265] 2.6 CTG detection

[0266] The cell culture plate was removed and allowed to equilibrate to room temperature for 30 minutes. 50 μL (equal to half the volume of the cell culture medium in each well) of CellTiter-Glo working solution was added to each well. The cell plate was wrapped with aluminum foil to avoid light. The culture plate was shaken on an orbital shaker for 2 minutes to induce cell lysis. The culture plate was placed at room temperature for 10 minutes to stabilize the luminescent signal, and the luminescent signal was detected on a 2104 EnVision plate reader.

[0267] 2.7 Data Analysis

[0268] The inhibition rate (IR) of the test compound was calculated using the following formula: IR (%) = (1 – (RLU compound – RLU blank control) / (RLU vehicle control – RLU blank control)) * 100%. Finally, nonlinear regression analysis was performed using the logarithm of compound concentration-inhibition rate in Graphpad Prism 5 software to obtain the IC value of the compound for inhibiting cell proliferation. 50 value.

[0269] The target compound claimed in the present invention has a good inhibitory effect on the growth of RET fusion engineered cells.

[0270] 3. In vivo antitumor activity of the compounds of the present invention:

[0271] Compounds with strong in vitro activity and low toxicity were selected to determine the maximum tolerated dose (MTD) in mice. The in vivo anti-tumor activity of the compounds of the present invention was determined in a nude mouse xenograft tumor model bearing human cancer to explore the dosage, route of administration, frequency, and cycle of administration required for the test compounds to produce pharmacodynamic effects.

[0272] 5-6 week old female BALB / C nude mice weighing approximately 18-20 g were raised. Human cancer xenograft tumor models were established in nude mice using the human colon cancer cell line colo205, the human breast cancer cell line MDA-MB435, and the human lung cancer cell line A549 obtained from ATCC. After digestion and detachment of the monolayer tumor cells, the cells were harvested and resuspended in serum-free culture medium at a concentration of 5 × 10 6 / 0.2mL, placed in an ice box and brought to the animal room, 0.2mL of cell suspension was directly taken with a syringe with a No. 6 needle and transplanted subcutaneously in the scapular area behind the left armpit of nude mice, 5×10 6 / 0.2mL / mouse, and measure the tumor volume every 2-3 days. After two weeks, select nude mice with tumors that are growing vigorously and without ulceration. Under sterile conditions, remove the tumors, cut the tumor tissue into pieces with a diameter of about 2-3mm, and inoculate them subcutaneously at the shoulder blade behind the left armpit of the nude mice. After three generations, when the tumor volume grows to 100mm, 3 Nude mice with tumors that were too large or too small were randomly divided into groups for drug administration.

[0273] The mice were randomly divided into five groups: a negative control (vehicle), a positive control (nintedanib, 4 mg / kg), and three treatment groups (20 mg / kg, 12 mg / kg, and 4 mg / kg, respectively, with the high dose below the MTD). Each group consisted of 8 nude mice, including 16 in the negative control group. The mice were intraperitoneally injected once weekly for 4 weeks. Body weights and tumor volumes were measured every 3 days, and the number of deaths was recorded. Twenty-four hours after the last dose, the animals were sacrificed, and tumor volume, weight, and body weight were measured. Tumor volume and body weight growth curves, tumor inhibition rate, and mortality rates were plotted. The relative tumor growth rate (T / C) (%) was calculated using the formula: TRTV / CRTV * 100%. (TRTV: treatment group RTV; CRTV: negative control RTV. Relative tumor volume (RTV) = Vt / V0, where V0 is the tumor volume at the time of group dosing and Vt is the tumor volume after dosing.) The in vivo anti-tumor efficacy of the compound of the present invention is relative tumor proliferation rate T / C (%) ≤ 40%, and the difference is statistically significant, indicating that the compound has obvious pharmacodynamic effect, as shown in Table 7, Figures 1 and 2.

[0274] Table 7. Evaluation of the antitumor efficacy of compound application example 1 on TT cell xenograft tumor model

[0275] Furthermore, the compound of the present invention has a smaller effect on the body weight of model mice and has a smaller toxicity.

[0276] The details of the specific embodiments described in the present invention are not to be construed as limiting thereof. Various synonyms and modifications may be made without departing from the spirit and scope of the present invention, and it is known that these synonymous embodiments are part of the present invention.

Claims

1. A 3-(amino)methyleneindoline derivative represented by the general formula (I), and its optical isomers or pharmaceutically acceptable salts thereof: In the above general formula (I), R 1 Selected from: hydrogen atom, alkyl, heteroalkyl, arylalkyl, C3-C14 cycloalkyl, aryl, heteroaryl, heteroarylalkyl, heterocycloalkyl, carboxylalkyl, carboxylcycloalkyl; any of the above groups independently may be unsubstituted or may be substituted by one or more substituents, and these substituents include but are not limited to halogen, isotope, amino, carboxyl, phenyl, benzyl, phenyloxy, =O, -CF3, haloalkyl, alkyl, alkenyl, alkynyl, hydroxyl, hydroxyalkyl, alkoxy, alkoxyalkyl, heteroalkyl, arylalkyl, cycloalkyl, aryl, heteroaryl, heteroarylalkyl, heterocycloalkyl, heterocycloalkenyl, alkoxyalkyl, alkenyloxy, alkynyloxy, amino, alkylamino, aminoalkyl, alkylaminocarbonyl, sulfonyl, alkylsulfonyl, alkylsulfinyl, or aminosulfonyl; the alkyl is a C1-C8 straight chain or branched alkyl; R 2 Selected from: hydroxy, alkyl, aryl, alkoxy, heteroalkoxy, heteroalkoxy, arylalkoxy, C3-C8 cycloalkoxy, aryloxy, heteroaryloxy, heteroarylalkoxy, heterocycloalkoxy, amino, alkylamino, heteroalkylamino, arylalkylamino, cycloalkylamino, arylamino, heteroarylamino, heteroarylalkylamino, heterocycloalkylamino; any of the above groups may be independently unsubstituted or may be substituted by one or more substituents, including but not limited to halogen, isotope, Amino, carboxyl, phenyl, benzyl, phenyloxy, =O, -CF3, haloalkyl, alkyl, alkenyl, alkynyl, hydroxyl, hydroxyalkyl, alkoxy, alkoxyalkyl, heteroalkyl, arylalkyl, cycloalkyl, aryl, heteroaryl, heteroarylalkyl, heterocycloalkyl, heterocycloalkenyl, alkoxyalkyl, alkenyloxy, alkynyloxy, amino, alkylamino, aminoalkyl, alkylaminocarbonyl, sulfonyl, alkylsulfonyl, alkylsulfinyl, or aminosulfonyl; the alkyl is a C1-C8 straight chain or branched alkyl; R 3 , R 4 Each of the following groups is independently selected from the group consisting of: hydrogen, halogen, isotope, hydroxyl, amino, carboxyl, alkyl, heteroalkyl, alkenyl, alkynyl, arylalkyl, C3-C14 cycloalkyl, aryl, heteroaryl, heteroarylalkyl, heterocycloalkyl, alkyloxy, heteroalkyloxy, arylalkoxy, cycloalkyloxy, aryloxy, heteroaryloxy, heteroarylalkoxy, heterocycloalkyloxy, amino, alkylamino, heteroalkylamino, arylalkylamino, cycloalkylamino, arylamino, heteroarylamino, heteroarylalkylamino, heterocycloalkylamino, carboxyl, carboxylalkylaminocarbonyl, cycloalkylaminocarbonyl, heteroarylaminocarbonyl, heteroarylaminocarbonyl; any of the above groups The groups may be independently unsubstituted or substituted with one or more substituents, including but not limited to halogen, isotope, amino, carboxyl, phenyl, benzyl, phenyloxy, =O, -CF3, haloalkyl, alkyl, alkenyl, alkynyl, hydroxyl, hydroxyalkyl, alkoxy, alkoxyalkyl, heteroalkyl, arylalkyl, cycloalkyl, aryl, heteroaryl, heteroarylalkyl, heterocycloalkyl, heterocycloalkenyl, alkoxyalkyl, alkenyloxy, alkynyloxy, alkylamino, aminoalkyl, alkylaminocarbonyl, sulfonyl, alkylsulfonyl, alkylsulfinyl, or aminosulfonyl; the alkyl group is a C1-C8 straight or branched chain alkyl group; R 5 Selected from: hydrogen atom, isotope, alkyl, heteroalkyl, arylalkyl, C3-C14 cycloalkyl, aryl, heteroaryl, heteroarylalkyl, heterocycloalkyl; any of the above groups independently may be unsubstituted or may be substituted by one or more substituents, and these substituents include but are not limited to halogen, isotope, amino, carboxyl, phenyl, benzyl, phenyloxy, =O, -CF3, haloalkyl, alkyl, alkenyl, alkynyl, hydroxyl, hydroxyalkyl, alkoxy, alkoxyalkyl, heteroalkyl, arylalkyl, cycloalkyl, aryl, heteroaryl, heteroarylalkyl, heterocycloalkyl, heterocycloalkenyl, alkoxyalkyl, alkenyloxy, alkynyloxy, alkylamino, aminoalkyl, alkylaminocarbonyl, sulfonyl, alkylsulfonyl, alkylsulfinyl, or aminosulfonyl; the alkyl is a C1-C8 straight chain or branched alkyl; R 6 Selected from: hydrogen atom, alkyl, heteroalkyl, arylalkyl, C3-C14 cycloalkyl, heteroarylalkyl, heterocycloalkyl; any of the above groups may be unsubstituted or substituted by one or more substituents, and these substituents include alkyl and alkoxy; the alkyl is a C1-C8 straight chain or branched alkyl; R 7 Selected from: absence, hydrogen atom, isotope, halogen, alkyl, heteroalkyl, arylalkyl, C3-C14 cycloalkyl, aryl, heteroaryl, heteroarylalkyl, heterocycloalkyl; alkoxy, heteroalkoxy, arylalkoxy, cycloalkyloxy, aryloxy, heteroaryloxy, heteroarylalkoxy, heterocycloalkyloxy, amino, alkylamino, heteroalkylamino, arylalkylamino, cycloalkylamino, arylamino, heteroarylamino, heteroarylalkylamino, heterocycloalkylamino, carboxyl, alkylaminocarbonyl, any of the above groups independently may be unsubstituted or may be substituted by one or more substituents, and these substituents include but are not limited to halogen, alkyl, alkoxy; the alkyl is a C1-C8 straight chain or branched alkyl; R 7 Can be linked to the C on the benzimidazole ring 4 -、C 5 -、C 6 - or C 7 - position; L 1 Selected from: covalent bond, alkylene, heteroalkylene, -C(O)-, -C(O)-C(O)--, -C(O)-NH-, alkylene-NH-, alkylene-C(O)-NH-, -C(O)-alkylene, -C(O)-NH-heteroalkylene, heteroalkylene-NH-C(O)-, heteroalkylene-NH-; the alkylene is a C1-C8 straight chain or branched alkyl; any of the above groups can be independently substituted by one or more substituents, and these substituents include hydrogen atoms and alkyl groups; L 2 Selected from: hydrogen, alkyl, cycloalkyl, bicyclic, bridged (tricyclic), spirocyclic, aryl, heteroaryl, heteroarylalkyl, heterocycloalkyl, alkoxy, heteroalkoxy, arylalkoxy, cycloalkyloxy, aryloxy, heteroaryloxy, heteroarylalkoxy, heterocycloalkoxy; any of the above groups may be independently unsubstituted or substituted by one or more substituents, including but not limited to halogen, isotope, amino, carboxyl, phenyl, benzyl, phenyloxy, =O, -CF3, haloalkyl, alkyl, alkenyl, alkynyl, hydroxyl, hydroxyalkyl, alkoxy, alkoxyalkyl, heteroalkyl, arylalkyl, cycloalkyl, aryl; the alkyl is a C1-C8 straight or branched chain alkyl; R 8 Selected from: absence, hydrogen atom, deuterium atom, hydroxyl, alkyl, hydroxyalkyl, heteroalkyl, alkenyl, alkynyl, arylalkyl, C3-C14 cycloalkyl, aryl, heteroaryl, arylalkyl, heteroarylalkyl, heterocycloalkyl, alkoxy, heteroalkoxy, arylalkoxy, cycloalkoxy, aryloxy, heterocycloalkoxy; any of the above groups may be independently unsubstituted or substituted with one or more substituents, including but not limited to halogen, isotope, amino, carboxyl, phenyl, benzyl, phenyloxy, =O, -CF3, haloalkyl, alkyl, alkenyl, alkynyl, hydroxyl, hydroxyalkyl, alkoxy, alkoxyalkyl, heteroalkyl, arylalkyl, cycloalkyl; the alkyl group is a C1-C8 straight chain or branched alkyl group; X is selected from: -CH(R 9 )-、-N(R 10 )-, -C(O)-; Y is selected from: a carbon atom, a nitrogen atom; R 9 and R 10 Each is independently selected from: a hydrogen atom, a deuterium atom, an alkyl group, a heteroalkyl group, and a C3-C14 cycloalkyl group; any of the above groups may be independently unsubstituted or substituted with one or more substituents, including but not limited to halogen, isotope, halogenated alkyl group, and alkyl group; the alkyl group is a C1-C8 straight chain or branched chain alkyl group; (R 6 )N can be linked to the C on the indoline ring 4 -、C 5 -、C 6 - and C 7 - position, in which case the general formula (I) is selected from:

2. The 3-(amino)methyleneindoline derivative according to claim 1, characterized in that: R 1 Selected from: hydrogen atom, alkyl, heteroalkyl, arylalkyl, C3-C14 cycloalkyl, aryl, heteroaryl, heteroarylalkyl, heterocycloalkyl, carboxylalkyl, carboxylcycloalkyl; any of the above groups independently may not be substituted or may be substituted by one or more substituents, and these substituents include but are not limited to halogen, isotope, amino, carboxyl, phenyl, benzyl, phenyloxy, =O, -CF3, haloalkyl, alkyl, alkenyl, alkynyl, hydroxyl, hydroxyalkyl, alkoxy, alkoxyalkyl, heteroalkyl, arylalkyl, cycloalkyl, heterocycloalkyl; the alkyl is a C1-C8 straight chain or branched alkyl.

3. The 3-(amino)methyleneindoline derivative according to claim 1, characterized in that: R 2 Selected from: hydroxy, alkyl, aryl, alkoxy, heteroalkoxy, heteroalkoxy, arylalkoxy, C3-C8 cycloalkoxy, heterocycloalkoxy, amino, alkylamino, heteroalkylamino, arylalkylamino, cycloalkylamino, arylamino, heteroarylamino, heteroarylalkylamino, heterocycloalkylamino; any of the above groups may be unsubstituted or substituted by one or more substituents, and these substituents include but are not limited to halogen, isotope, amino, carboxyl, alkyl, alkoxy, alkoxyalkyl, heteroalkyl; the alkyl is a C1-C8 straight chain or branched alkyl.

4. The 3-(amino)methyleneindoline derivative according to claim 1, characterized in that: R 2 Selected from: methoxy, ethoxy, propoxy, isopropoxy, hydroxy, butoxy, difluoroethoxy, D3CD2CO-, D3CO-.

5. The 3-(amino)methyleneindoline derivative according to claim 1, characterized in that: R 3 , R 4 are independently selected from the group consisting of: hydrogen, halogen, isotope, hydroxyl, amino, carboxyl, alkyl, heteroalkyl, alkenyl, alkynyl, arylalkyl, C3-C14 cycloalkyl, aryl, heteroaryl, heteroarylalkyl, heterocycloalkyl, alkoxy, heteroalkoxy, aryloxy, cycloalkyloxy, aryloxy, heteroaryloxy, heteroarylalkoxy, heterocycloalkyloxy, amino, alkylamino, heteroalkylamino, arylalkylamino, cycloalkylamino, arylamino, heteroarylamino, heteroarylalkylamino, heterocycloalkylamino, carboxyl, carboxylalkylaminocarbonyl, cycloalkylaminocarbonyl, heteroarylaminocarbonyl, heteroarylaminocarbonyl; any of the above groups Each of the groups independently may be unsubstituted or may be substituted with one or more substituents, which substituents include but are not limited to halogen, isotope, amino, carboxyl, phenyl, benzyl, phenyloxy, =O, -CF3, haloalkyl, alkyl, alkenyl, alkynyl, hydroxyl, hydroxyalkyl, alkoxy, alkoxyalkyl, heteroalkyl, arylalkyl, cycloalkyl, aryl, heteroaryl, heteroarylalkyl, heterocycloalkyl, heterocycloalkenyl, alkoxyalkyl, alkenyloxy, alkynyloxy, alkylamino, aminoalkyl, alkylaminocarbonyl, sulfonyl, alkylsulfonyl, alkylsulfinyl, or aminosulfonyl; the alkyl group is a C1-C8 straight chain or branched alkyl group.

6. The 3-(amino)methyleneindoline derivative according to claim 1, characterized in that: R 4 Selected from: cyclopropane, cyclobutane, cyclopentane, methyl, ethyl, propyl, isopropyl, isobutyl, -CH(CH3)CH2CH3.

7. The 3-(amino)methyleneindoline derivative according to claim 1, characterized in that: R 5 Selected from: hydrogen atom, isotope, alkyl, heteroalkyl, arylalkyl, C3-C14 cycloalkyl, aryl, heteroaryl, heteroarylalkyl, heterocycloalkyl, heteroalkyl; any of the above groups independently may be unsubstituted or may be substituted by one or more substituents, and these substituents include but are not limited to halogen, isotope, amino, carboxyl, phenyl, benzyl, phenyloxy, =O, -CF3, haloalkyl, alkyl, alkenyl, alkynyl, hydroxyl, hydroxyalkyl, alkoxy, alkoxyalkyl, heteroalkyl, arylalkyl, cycloalkyl, aryl, heteroaryl, heteroarylalkyl, heterocycloalkyl, heterocycloalkenyl, alkoxyalkyl, alkenyloxy, alkynyloxy, alkylamino, aminoalkyl, alkylaminocarbonyl, sulfonyl, alkylsulfonyl, alkylsulfinyl, or aminosulfonyl; the alkyl is a C1-C8 straight chain or branched alkyl.

8. The 3-(amino)methyleneindoline derivative according to claim 1, characterized in that: R 5 Selected from: hydrogen atom, benzene ring, methyl group, ethyl group, propyl group.

9. The 3-(amino)methyleneindoline derivative according to claim 1, characterized in that: R 6 Selected from: a hydrogen atom, an alkyl group, a C3-C14 cycloalkyl group.

10. The 3-(amino)methyleneindoline derivative according to claim 1, characterized in that: R 7 Selected from: absence, hydrogen atom, isotope, halogen, alkyl, heteroalkyl, C3-C14 cycloalkyl, aryl, heteroaryl, alkoxy, heteroalkoxy, aryloxy, heteroaryloxy, heterocycloalkoxy, amino, alkylamino, heteroalkylamino, carboxyl, alkylaminocarbonyl; any of the above groups can be substituted by one or more substituents, including but not limited to halogen, alkyl, alkoxy; the alkyl is a C1-C8 straight chain or branched alkyl.

11. The 3-(amino)methyleneindoline derivative according to claim 1, characterized in that: R 7 Selected from: Fluorine.

12. The 3-(amino)methyleneindoline derivative according to claim 1, characterized in that: L 1 Selected from: covalent bond, alkylene, heteroalkylene, -C(O)-, -C(O)-C(O)-, -C(O)-NH-, alkylene-NH-, alkylene-C(O)-NH-, -C(O)-alkylene, -C(O)-NH-heteroalkylene, heteroalkylene-NH-C(O)-, heteroalkylene-NH-; the alkylene is a C1-C8 straight chain or branched alkyl; any of the above groups can be independently substituted by one or more substituents, and these substituents include hydrogen atoms and alkyl groups.

13. The 3-(amino)methyleneindoline derivative according to claim 1, characterized in that: L 2 Selected from: hydrogen atom, alkyl, heteroalkyl, arylalkyl, C3-C14 cycloalkyl, aryl, heteroaryl, heterocycloalkyl, alkoxy, heteroalkoxy, cycloalkyloxy, heterocycloalkyloxy, amino, alkylamino, heteroalkylamino, arylalkylamino, cycloalkylamino, arylamino; any of the above groups independently may be unsubstituted or may be substituted by one or more substituents, and these substituents include but are not limited to halogen, isotope, amino, carboxyl, phenyl, benzyl, phenyloxy, =O, -CF3, haloalkyl, alkyl, alkenyl, alkynyl, hydroxyl, hydroxyalkyl, alkoxy, alkoxyalkyl, heteroalkyl, arylalkyl, cycloalkyl, aryl, heteroaryl, heteroarylalkyl, heterocycloalkyl, heterocycloalkenyl, alkoxyalkyl, alkenyloxy, alkynyloxy, alkylamino; the alkyl is a C1-C8 straight chain or branched alkyl.

14. The 3-(amino)methyleneindoline derivative according to claim 1, characterized in that: X is selected from: a carbon atom, a carbonyl group.

15. The 3-(amino)methyleneindoline derivative according to claim 1, characterized in that: R 8 Selected from: absent, hydrogen atom, hydroxyl, alkyl, heteroalkyl, alkenyl, alkynyl, arylalkyl, C3-C14 cycloalkyl, heterocycloalkyl, alkoxy, heteroalkoxy, arylalkoxy, cycloalkyloxy, aryloxy, heteroaryloxy, heteroarylalkoxy, heterocycloalkyloxy; any of the above groups independently may be unsubstituted or substituted by one or more substituents, and these substituents include but are not limited to halogen, isotope, amino, carboxyl, =O, -CF3, haloalkyl, alkyl, alkenyl, alkynyl, hydroxyl, hydroxyalkyl, alkoxy, alkoxyalkyl, heteroalkyl, arylalkyl, cycloalkyl, aryl, heteroaryl, heteroarylalkyl, heterocycloalkyl, heterocycloalkenyl, alkoxyalkyl, alkenyloxy, alkynyloxy, alkylamino, aminoalkyl; the alkyl is a C1-C8 straight chain or branched alkyl.

16. The 3-(amino)methyleneindoline derivative according to claim 1, characterized in that: R 8 It is methyl.

17. The 3-(amino)methyleneindoline derivative according to claim 1, characterized in that: R 8 It is a disubstituted methyl group at the 3-substitution position.

18. The 3-(amino)methyleneindoline derivative according to any one of claims 1 to 17, wherein the structure thereof can be selected from one of the following structures, or a stereoisomer, geometric isomer, tautomer, nitrogen oxide, solvate, metabolite, pharmaceutically acceptable salt or prodrug thereof:

19. The method for preparing 3-(amino)methyleneindoline derivatives according to any one of claims 1 to 18, characterized in that: When (R 6 )N can be linked to the C on the indoline ring 5 - position, and R 1 =R 3 =R 4 =R 6 =H, R 2 =-OCH3, the preparation method of the 3-(amino)methyleneindoline derivatives comprises the following steps: S1, reacting a substituted 5-nitro-indoline derivative (VI) with a chlorinated compound (VII) to obtain a 5-nitro-substituted indoline derivative (VIII); S2, 5-nitro-substituted indoline derivative (VIII) is reacted with active hydrogen-containing compound (IX) under heating in toluene to obtain compound (X); S3, compound (X) is reduced to obtain (XI), which is then reacted with 3-methoxymethylene-oxoindoline-6-carboxylate (XII) to obtain the target compound represented by the general formula (I), i.e., (XIII); The synthetic route is as follows:

20. The method for preparing 3-(amino)methyleneindoline derivatives according to any one of claims 1 to 11, characterized in that: When (R 6 )N can be linked to the C on the benzimidazole ring 5 - position, and R 1 =R 3 =R 5 =R 6 =H, R 2 =OCH3 and X=-C(O)-, the preparation method of the 3-(amino)methyleneindoline derivatives comprises the following steps: S1. Substituted 4-bromo-2-oxoindoline-6-carboxylic acid methyl ester (XIV) reacts with a boron derivative (XV) in the presence of a palladium reagent to obtain a compound (XVI); S2. Using a suitable acid as solvent, compound (XVI) reacts with triethyl orthoformate to obtain compound (XVII); S3, reacting a 5-nitroindole derivative (XVIII) with a suitable compound (XIX) under base catalysis to obtain a compound (XX); S4, compound (XX) is reduced to obtain an important intermediate (XXI); S5. Compound (XVII) undergoes substitution reaction with intermediate (XXI) to obtain target compound (XXII) represented by general formula (I); The synthetic route is as follows:

21. Any pharmaceutical dosage form comprising the 3-(amino)methyleneindoline derivative according to any one of claims 1 to 19 and a pharmaceutically acceptable diluent, excipient or carrier.

22. The use of the compound according to any one of claims 1 to 19, characterized in that The invention is used for preparing a drug for treating a disease caused by, associated with or accompanied by the disruption of cell proliferation and / or angiogenesis, or for inhibiting kinase activity.

23. The use of the compound according to claim 23, characterized in that: The disorder is a proliferative disease.

24. The use of the compound according to claim 24, characterized in that The proliferative disease is cancer.

25. The use of the compound according to claim 23, characterized in that Inhibiting kinase activity includes inhibiting the activity of RET, PDGFR, VEGFR, FGFR, FLT3, Aurora-A, Aurora-B, TRK, RAF, RET or Abl.

26. The use of the compound according to claim 23, characterized in that The disease is selected from: bone cancer, including: Ewing's sarcoma, osteosarcoma, chondrosarcoma, etc.; brain and CNS tumors, including: acoustic neuroma, neuroblastoma, glioma and other brain tumors, spinal cord tumors, breast cancer, colorectal cancer, advanced colorectal adenocarcinoma; endocrine cancer, including: adrenal cortical carcinoma, pancreatic cancer, pituitary cancer, thyroid cancer, parathyroid cancer, thymic cancer, multiple endocrine tumors; gastrointestinal cancer, including: gastric cancer, esophageal cancer, small intestine cancer, liver cancer, Cancer, extrahepatic bile duct cancer, gastrointestinal carcinoid tumors, gallbladder cancer; genitourinary cancers, including: testicular cancer, penis cancer, prostate cancer; gynecological cancers, including: cervical cancer, ovarian cancer, vaginal cancer, uterine / endometrial cancer, pudendal cancer, gestational trophoblastic tumor, fallopian tube cancer, uterine sarcoma; head and neck tumors, including: oral cancer, lip cancer, salivary gland cancer, laryngeal cancer, hypopharyngeal cancer, orthopharyngeal cancer, nasal cancer, paranasal sinus cancer, nasopharyngeal cancer; blood cancers, including: childhood leukemia, Acute lymphocytic leukemia, acute myeloid leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, hairy cell leukemia, acute promyelocytic leukemia, plasma cell leukemia; bone marrow cancer blood diseases, including: myelodysplastic syndrome, myeloproliferative disorders, aplastic anemia, Fanconi anemia, idiopathic macroglobulinemia; lung cancer, including: small cell lung cancer, non-small cell lung cancer; lymphoma, including: Hodgkin's disease, non-Hodgkin's lymphoma, cutaneous T-cell lymphoma, peripheral T-cell lymphoma, AIDS-related lymphoma; eye cancers, including retinoblastoma, uveal melanoma; skin cancers, including melanoma, non-melanoma skin cancer, Merkel cell carcinoma; soft tissue sarcomas, such as childhood soft tissue sarcoma, adult soft tissue sarcoma, Kaposi sarcoma; urinary system cancers, including renal cancer, Wilms' tumor, bladder cancer, urethral cancer and metastatic cell carcinoma.

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