A pyrimidine derivative and its preparation method and application

Abstract

The present invention discloses a novel pyrimidine derivative, a preparation method thereof and a medical application thereof. More specifically, these compounds can be used as drugs for treating proliferative diseases and other diseases associated with abnormal expression of various kinases. In particular, these compounds can be used as drugs for treating proliferative diseases and other diseases associated with abnormal expression of various kinases such as RET, TRK, FGHR, PDGFR, VEGFR, etc. The structure of the novel pyrimidine of the present invention is shown in the general formula (I), wherein R 1 ,R 2 ,R 3 , L 1 ,L 2 The definitions of and X are given in the specification.

Description

Technical Field

[0001] The present invention relates to a novel pyrimidine derivative, a preparation method and application thereof. Specifically, it relates to a preparation method of a novel pyrimidine derivative having the ability to inhibit the growth of wild tumor cells with high kinase expression or tumor cell lines with corresponding kinase mutations. In particular, these compounds can be used as drugs for treating proliferative diseases and other diseases related to abnormal expression of various kinases such as RET, FGHR, PDGFR, VEGFR, etc. In addition, the present invention relates to drugs containing these compounds and the application of these compounds in the preparation of drugs. Background Art

[0002] Tumor is one of the major diseases that threaten human health. Chemotherapy is still one of the important means of tumor treatment. In particular, small molecule targeted therapy for tumors and combined therapy with other methods are playing an increasingly important role. Among the many targeted therapy drugs, inhibitors acting on RET, TRK, FGHR, PDGFR, and VEGFR have received widespread attention.

[0003] RET is a neuronal growth factor receptor tyrosine kinase and a transmembrane glycoprotein. RET is expressed by the proto-oncogene RET (REarranged durIng TransfectIon) located on chromosome 10. It plays an important role in the development of the kidney and enteric nervous system in the embryonic stage. It is also critical for the homeostasis of multiple tissues, such as neurons, neuroendocrine, hematopoietic tissues and male germ cells.

[0004] Unlike other RTKs, RET does not bind directly to ligand molecules: such as artemin, glial cell-derived neurotrophic factor (GDNF), neurturin and persephin, which are all GNDF family ligands (GFLs). These ligand GFLs usually bind to GDNF family receptor α (GFRα), and 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, activating cell proliferation and other signal transduction cascade reactions, and related signaling pathways include MAPK, PI3K, JAK-STAT, PKA, PKC, etc. ((1)Maria Grazia Borrello,Elena Ardini,Laura D Locati,Angela Greco,LisaLicitra&Marco A PIerotti(2013).RET Inhibition:ImplicatIons in cancertherapy.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 ClinEndocrinol Metab 98:3149–3164, 2013).

[0005] There are two main mechanisms of RET oncogenic activation: one is that chromosomal rearrangement produces a new fusion protein, usually a fusion of the RET kinase domain and a protein containing a self-dimerization domain; the other is that RET mutations directly or indirectly activate the kinase activity of RET. These changes at the somatic or germ cell level are involved in the pathogenesis of various cancers. 5%-10% of patients with papillary thyroid cancer have RET chromosomal rearrangements; 60% of patients with medullary thyroid carcinoma have RET point mutations; among all NSCLC patients, approximately 1-2% have RET fusion proteins, of which KIF5B-RET is more common.

[0006] These facts all indicate an ideal treatment for tumors associated with persistent RET activation. The research on RET inhibitors has received widespread attention and has made rapid progress. Among them, RET inhibitors represented by Pralsetinib (WO2017011776A1 ArrayLoxo 292) and SelpercatInIb (WO2017079140A1 Blu667) have been approved by the FDA for marketing and have been used to treat various related fusion tumors regardless of tumor type and have been successful. This has inspired various other development attempts. ((1)Lucille Lopez-Delisle,Cécile Pierre-Eugène,CarolineLouis-Brennetot,Didier Surdez,Virginie Raynal,Sylvain Baulande,Valentina Boeva,Sandrine Grossetête-Laiami,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 andGastrointestinal Disorders.(3)MInsoo Song.Progress In Discovery of KIF5B-RETKinase Inhibitors for the Treatment of Non-Small-Cell Lung Cancer.Miniperspective.J.Med.Chem.2015,58,3672-3681.)

[0007]

[0008] As mentioned above, the research and development of kinase inhibitors targeting RET have achieved great success in the treatment of fusion tumors, bringing good news to the majority of patients. However, the proportion of tumors with RET as the core and fusion with other genes is still small (Alexander Drilon, Zishuo I. Hu, Gillianne GY Lai & Daniel S. W. Tan Targeting RET-driven cancers: lessons from evolving preclinical and clinical landscapes. Nature Reviews Clinical Oncology volume 15, 2018, 151–167). Obviously, it would be an effective option if various gene fusions of RET could be effectively inhibited at the same time. On the other hand, for almost all kinase targets, drug resistance is always the main problem. Therefore, there is still a huge social demand for more RET, TRK, FGHR, PDGFR, and VEGFR kinase inhibitors in this field. Summary of the invention

[0009] One of the purposes of the present invention is to disclose a novel pyrimidine derivative. This type of compound acts on RET, TRK, FGHR, PDGFR, VEGFR, and can be used to treat tumors, endocrine disorders, immune system diseases, genetic diseases, and neurodegenerative diseases.

[0010] The second purpose of the present invention is to disclose a method for preparing the pyrimidine derivatives.

[0011] More specifically, the present invention relates to pyrimidine-containing organisms and methods for preparing the same; these compounds can be used as drugs for treating proliferative disorders and other diseases associated with abnormal expression of various kinases. The structures thereof are shown in general formula (I) and formula (I), wherein R 1 ,R 2 ,R 3 ,L 1 ,L 2 ,L 3 and X is defined as follows:

[0012]

[0013] In the general formula (I)

[0014] R 1is selected from the group consisting of: a hydrogen atom, an alkyl group, a heteroalkyl group, an alkenyl group, an alkynyl group, an arylalkyl group, a C3-C8 cycloalkyl group, an aryl group, a heteroaryl group, a heteroarylalkyl group, a heterocycloalkyl group, a bicyclic group, a bridged ring (tricyclic) group, a spirocyclic group, an alkyl ether group, a heteroalkyl ether group, an arylalkyl ether group, a cycloalkyl ether group, an aryl ether group, a heteroaryl ether group, a heteroarylalkyl ether group, a heterocycloalkyl ether group, an alkylamino group, a heteroalkylamino group, an arylalkylamino group, a cycloalkylamino group, an arylamino group, a heteroarylamino group, a heteroarylalkylamino group, a heterocycloalkylamino group, a carboxyl group, a carboxylalkylaminocarbonyl group, a cycloalkylaminocarbonyl group, a heteroarylaminocarbonyl group, a heteroarylaminocarbonyl group; any of the above groups The alkyl radicals are 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 radical is a C1-C8 straight or branched chain alkyl radical;

[0015] Y is selected from: alkylene, heteroalkylene, carbonyl, -O-, -NH-, -CO-NH-, -NH-CO-, -alkyl-CO-NH-, -alkyl-NH-CO-, -alkyl-NH-, -alkyl-O-, nitrogen oxide six-membered ring; the alkyl group is a C1-C8 straight chain or branched alkyl group; any of the above groups can be substituted by one or more substituents independently, and these substituents include hydrogen atoms and alkyl groups;

[0016] R 2 Selected from: hydrogen atom, alkyl, heteroalkyl, alkenyl, alkynyl, arylalkyl, cycloalkyl, aryl, heteroaryl, heteroarylalkyl, heterocycloalkyl, alkyl ether, heteroalkyl ether, arylalkyl ether, cycloalkyl ether, aryl ether, heteroaryl ether, heteroaryl alkyl ether, heterocycloalkyl ether, carboxyl, carboxylalkylaminocarbonyl, cycloalkylaminocarbonyl, heteroarylaminocarbonyl, heteroarylaminocarbonyl; any of the above groups may be independently unsubstituted or substituted with one or more substituents, including but not limited to halogen, isotope, amino, hydroxyl, hydroxyalkyl, alkoxy, alkoxyalkyl, heteroalkyl, cycloalkyl, aryl, heteroaryl, heterocycloalkyl, heterocycloalkenyl, alkoxyalkyl, alkylamino, aminoalkyl; the alkyl group is a C1-C8 straight chain or branched alkyl group;

[0017] L 1 Selected from: -CH2CH2-, -CH=CH-, -C≡C-;

[0018] L 2Selected from: alkylene, heteroalkylene, carbonyl, -C(O)-NH-, alkyl-C(O)-NH-, alkyl-NH-C(O)-, -N(R 7 )H-,-R 8 -O-, -C(O)-; R 7 and R 8 Each of the above groups is independently selected from C1-C5 straight chain or branched alkyl groups; any of the above groups can be substituted by one or more substituents, including hydrogen atoms and alkyl groups;

[0019] R 3 is selected from the group consisting of: a hydrogen atom, an alkyl group, a heteroalkyl group, an alkenyl group, an alkynyl group, an arylalkyl group, a cycloalkyl group, an aryl group, a heteroaryl group, a heteroarylalkyl group, a heterocycloalkyl group, a bicyclic group, a bridged ring (tricyclic) group, a spirocyclic group, an alkyl ether group, a heteroalkyl ether group, an arylalkyl ether group, a cycloalkyl ether group, an aryl ether group, a heteroaryl ether group, a heteroarylalkyl ether group, a heterocycloalkyl ether group, a carboxyl group, a carboxylalkylaminocarbonyl group, a cycloalkylaminocarbonyl group, a heteroarylaminocarbonyl group, and a heteroarylaminocarbonyl group; any of the above groups may be independently unsubstituted or may be substituted by one or more substituents, and these substituents may be substituted by one or more substituents. Substituted groups 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.

[0020] In certain embodiments, in the above general formula (I), R 1 Selected from alkyl, heteroalkyl, arylalkyl, cycloalkyl, heteroarylalkyl, heterocycloalkyl, bicyclic, bridged (tricyclic), spirocyclic, alkoxyalkyl, aminoalkyl, alkylsulfonyl, alkylsulfinyl, aminosulfonyl, acyl; 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.

[0021] In certain embodiments, in the above general formula (I), R 1 Selected from R 5 -OH; R 4 Selected from C1-C3 alkyl, C3-C8 cycloalkyl; R 5 It is a C1-C5 straight chain or branched chain alkyl.

[0022] In certain embodiments, in the above general formula (I), R 2 It is a C1-C5 straight chain or branched chain alkyl.

[0023] In certain embodiments, in the above general formula (I), R 3 Selected from

[0024] Benzene ring, substituted benzene ring, pyridine, substituted pyridine, pyrimidine, substituted pyrimidine, CH-(CH2)nR 9 The substituted benzene ring contains 1-3 substituents, and the substituents are selected from halogen, C1-C3 alkyl, methyl ether, trifluoromethyl ether, ethyl ether, trifluoroethyl ether, propyl ether, trifluoropropyl ether, trifluoromethyl, amino, The substituted pyridine contains 1-3 substituents, and the substituents are selected from halogen, C1-C3 alkyl, methyl ether, trifluoromethyl ether, ethyl ether, trifluoroethyl ether, propyl ether, trifluoropropyl ether, trifluoromethyl, The substituted pyrimidine contains 1 to 3 substituents selected from Amino;

[0025] R 9 for Benzene ring, substituted benzene ring, pyridine, substituted pyridine, methyl ether group, ethyl ether group, propyl ether group, Pyrimidine, substituted pyrimidine;

[0026] R 9 The substituted benzene ring contains 1-3 substituents selected from halogen, C1-C3 alkyl, methyl ether, trifluoromethyl ether, ethyl ether, trifluoroethyl ether, propyl ether, trifluoropropyl ether, trifluoromethyl,

[0027] R 9 The substituted pyridine contains 1-3 substituents selected from halogen, C1-C3 alkyl, methyl ether, trifluoromethyl ether, ethyl ether, trifluoroethyl ether, propyl ether, trifluoropropyl ether, trifluoromethyl, R 9 The substituted pyrimidine contains 1 to 3 substituents selected from Amino;

[0028] R 10 Selected from hydrogen,

[0029] In certain embodiments, in the above general formula (I), L1 is: -CH=CH-.

[0030] In certain embodiments, in the above general formula (I), L1 is: -C≡C-.

[0031] In certain embodiments, in the above general formula (I), L2 is: alkylene.

[0032] In certain embodiments, in the above general formula (I), L2 is: carbonyl.

[0033] In certain embodiments, in the above general formula (I), L2 is: -CO-NH-.

[0034] In certain embodiments, in the above general formula (I), L2 is: -alkyl-NH-.

[0035] In certain embodiments, in the above general formula (I), L2 is: -alkyl-O-.

[0036] In certain embodiments, in the above general formula (I), Y is: alkylene, carbonyl, -O-, -NH-, -CO-NH-, -NH-CO-, -alkyl-CO-NH-, -alkyl-NH-CO-, -alkyl-NH-, -alkyl-O-.

[0037] In certain embodiments, in the above general formula (I), R2 is: hydrogen atom, alkyl, heteroalkyl, alkenylalkyl, aryl, heteroaryl, heterocycloalkyl, alkyl ether, aryl ether, heteroaryl ether, heteroaryl alkyl ether, heterocycloalkyl ether, carboxyl; 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, 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.

[0038] In certain embodiments, in the above general formula (I), R 3 It is alkyl, heteroalkyl, cycloalkyl, heteroaryl, heteroarylalkyl, heterocycloalkyl, bicyclic, bridged ring (tricyclic) radical, spirocyclic radical. Any of the above groups may be independently substituted or may be substituted by one or more substituents, and these substituents include but are not limited to halogen, isotope, amino, =O, -CF3, haloalkyl, alkyl, hydroxyl, hydroxyalkyl, alkoxy, alkoxyalkyl, heteroalkyl, arylalkyl, cycloalkyl, aryl, heteroaryl, heteroarylalkyl, heterocycloalkyl.

[0039] In some embodiments, the compounds described herein have one of the following structures, or a stereoisomer, geometric isomer, tautomer, nitrogen oxide, solvate, metabolite, pharmaceutically acceptable salt, or prodrug thereof:

[0040]

[0041]

[0042]

[0043]

[0044] The present invention also discloses a method for preparing the above-mentioned pyrimidine derivatives. When Y=NH, L1=CH=CH, and L2=CO-NH, the preparation method comprises:

[0045] S1. converting the substituted 6-chloro-2-aminopyrimidine derivative (II) into the corresponding 6-chloro-2-iodopyrimidine derivative (III);

[0046] S2, 6-chloro-2-iodopyrimidine derivative (III) and amino derivative (I) are condensed to obtain compound (IV);

[0047] S3. Under amino protection, compound (IV) generates intermediate (V), which undergoes Heck reaction with a suitable acrylate to be converted into compound (VII);

[0048] S4, alkaline hydrolysis of compound (VII), condensation of the obtained acid (VIII) with the amino compound (IX) to obtain compound X;

[0049] S5. Compound X is hydrolyzed to remove the protecting group, thereby obtaining compound XI, i.e., the target compound represented by general formula (I);

[0050] The structures of the compounds involved include:

[0051]

[0052] In certain embodiments, the preparation method of the above-mentioned pyrimidine derivatives has the following synthetic route:

[0053] Synthetic route 1

[0054]

[0055] In certain embodiments, when R 1 When it is 5-methyl-1H-pyrazole, the corresponding R 1-NH2 can be synthesized by the method shown in Synthesis Scheme 2: the nitro compound (XII) can be reduced under protection to obtain the corresponding amino compound I-1 (see Synthesis Scheme 1 for details)

[0056] Synthetic route 2

[0057]

[0058] 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.

[0059] 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.

[0060] 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.

[0061] The present invention provides the use of an effective amount of a compound represented by the general formula (I) alone or in combination with other drugs in the preparation of a drug for treating a disease caused by, associated with or accompanied by the destruction of cell proliferation and / or angiogenesis.

[0062] In certain embodiments, the disorder is a proliferative disease.

[0063] In certain embodiments, the proliferative disease is cancer.

[0064] The present invention also includes the use of the compound represented by the above general formula (I) or its pharmaceutically acceptable salt or the above pharmaceutical composition for preparing a drug for inhibiting the activity of kinase.

[0065] In certain embodiments, the inhibition of kinase activity is inhibition of RET, TRK, FGHR, PDGFR, VEGFR activity.

[0066] The present invention also includes a drug for treating a patient's condition caused by, or associated with, or accompanied by the destruction of cell proliferation and / or angiogenesis, the drug 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.

[0067] The present invention also includes a drug for treating a disease that can be treated by inhibiting kinase in a patient, the drug comprising administering to the patient a therapeutically effective amount of the compound represented by the above general formula (I) or a pharmaceutically acceptable salt thereof.

[0068] 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 carcinoma, multiple endocrine tumors; gastrointestinal cancers , including: gastric cancer, esophageal cancer, small intestine cancer, kidney cancer, renal cell carcinoma, liver 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 cancers, including: oral cancer, lip cancer, salivary gland cancer, laryngeal cancer, hypopharyngeal cancer, orthopharyngeal cancer, nasal cancer, paranasal sinus cancer, nasal Pharyngeal cancer; blood cancer, 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, Fanwohn anemia, idiopathic macroglobulinemia; lung cancer, including: small cell lung cancer, non-small cell lung cancer; lymphoma, including: Hodgkin's disease, 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.

[0069] 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

[0070] Figure 1 The anti-tumor inhibitory activity of compound 1 in the TT model. DETAILED DESCRIPTION

[0071] The present invention discloses a preparation method and medical application of novel pyrimidine derivatives. These compounds can be used as, but not limited to, kinase inhibitors. The aminopyrazole derivatives disclosed in the present invention can be used alone or together with other drugs or pharmaceutically acceptable carriers, diluents or excipients, and are suitable for preventing or treating diseases caused by, associated with or accompanied by the destruction of cell proliferation and / or angiogenesis. One example of these diseases is cancer.

[0072] As used herein, the term "cancer" generally refers to a broad range of disorders characterized by the uncontrolled abnormal growth of cells.

[0073] The compounds of the present invention are expected to be useful in treating a variety of 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: adrenal cortical carcinoma, pancreatic cancer, pituitary cancer, thyroid cancer, parathyroid cancer, thymic carcinoma, multiple endocrine tumors; gastrointestinal cancers, including: : Gastric cancer, esophageal cancer, small intestine cancer, kidney cancer, renal cell carcinoma, liver 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 cancer, 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 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.

[0074] The compounds disclosed in the present invention can be used to treat cancers including: breast cancer, lung cancer, ovarian cancer, prostate cancer, head and neck cancer, kidney, stomach and brain cancer.

[0075] The compounds disclosed in the present invention can be used to treat the following cancers: cutaneous T-cell lymphoma (CTCL) and peripheral T-cell lymphoma.

[0076] Preferred cancers that can be treated by the compounds of the invention are solid tumors and hematological malignancies.

[0077] The compounds may also be used to treat disorders involving, relating to, or associated with dysregulation of histone deacetylases. There are many disorders known to involve or be regulated at least in part by HDAC activity, where HDAC activity is known to play a role in promoting the onset of the disease, or where the symptoms are known or have been shown to be alleviated by HDAC inhibitors. Disorders of this type that are expected to be treated by the compounds of the invention include, but are not limited to, the following: antiproliferative disorders (e.g., cancer); neurodegenerative diseases, including Huntington's disease, polyglutamine disease, Parkinson's disease, Alzheimer's disease, epileptic seizures, striatonigral degeneration, progressive supranuclear palsy, torsion dystonia, spastic torticollis and dyskinesia, familial tremor, Tourette syndrome, diffuse Lewy body disease, progressive supranuclear palsy, Pick's disease, intracranial hemorrhage, original 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, rubrum iridophylloglaucoma; inflammatory diseases and / or immune system disorders, including: rheumatoid arthritis (RA), osteoarthritis, juvenile chronic arthritis, graft-versus-host disease, psoriasis, asthma, spondyloarthropathies, psoriasis, Crohn's disease, inflammatory bowel disease, colon ulcers, alcoholic hepatitis, diabetes, Sjoegrens syndrome, multiple sclerosis, ankylosing spondylitis, membranous glomerulopathy, disc pain, systemic lupus erythematosus; diseases involving angiogenesis, including: cancer, psoriasis, rheumatoid arthritis; psychological disorders, including: bipolar disorder, 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, bacterial infections; viral infections, such as: herpes simplex; protozoan infections, such as: malaria, Leishmania infection, Trypanosoma brucei infection, toxoplasmosis and coccidiosis; hematopoietic disorders, including: thalassemia, anemia and sickle cell anemia.

[0078] The term "unsubstituted" as used herein means no substituent or substitution with only hydrogen.

[0079] Some of the terms used in this invention are defined as follows:

[0080] "Halogen" refers to fluorine, chlorine, bromine and iodine.

[0081] "=O" means oxo.

[0082] "-CF3" refers to trifluoromethyl.

[0083] "-CO-NH-" is "-amide-".

[0084] "-NH-CO-" is "-aminoacyl-".

[0085] "Carbonyl" means

[0086] "Alkyl" when used as a group or part of a group refers to a straight chain or branched aliphatic hydrocarbon group. Preferably, the alkyl group is C1-C14 alkyl; more preferably, C1-C10 alkyl; most preferably, C1-C6 alkyl, unless otherwise specified. 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.

[0087] "Cycloalkyl" refers to a saturated or partially saturated monocyclic, condensed or spirocyclic carbon ring. Rings consisting of 3-9 carbon atoms are preferred. Examples include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, etc.

[0088] "Heteroalkyl" refers to a group containing a straight chain or a branched chain alkyl group, and in the main chain, contains at least one or more heteroatoms selected from S, O and N. Preferably, the chain contains 2-14 atoms. Heteroalkyl includes, but is not limited to, ethers, thioethers, alkyl esters, second or third alkylamines, alkyl sulfinic acids, etc.

[0089] "Heterocycloalkyl" refers to a group formed by replacing one or more (preferably 1, 2 or 3) carbon atoms in the "cycloalkyl" defined above with oxygen, nitrogen, phosphorus, boron, selenium, silicon or sulfur atoms (preferably oxygen, sulfur or nitrogen). Wherein, heterocycloalkyl and alkyl parts are defined in this document. Preferably, it contains 1-3 heteroatoms. The preferred ring is a 3-14-membered ring (i.e., a 3-14-membered heterocycloalkyl), and the more preferred ring is a 4-7-membered ring (i.e., a 4-7-membered heterocycloalkyl). Heterocycloalkyl includes, but is 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. Heterocycloalkyl may be substituted by one or more substituents.

[0090] "Bicyclic group" refers to a "cycloalkyl" or "heterocycloalkyl" group as defined above in which at least two atoms in the ring system are common atoms of the bicyclic ring.

[0091] The bridged cyclo(tricyclic) group refers to a "cycloalkyl" or "heterocycloalkyl" ring system as defined above, in which at least one atom is a bridge atom.

[0092] Spirocyclyl refers to a ring having at least one atom which is a common atom in a bicyclic ring.

[0093] "Heterocycloalkylalkyl" refers to a group of (heterocycloalkyl-alkyl)-, wherein the heterocycloalkyl and alkyl parts are as defined herein. Heterocycloalkylalkyl groups include, but are not limited to, (2-tetrahydrofuranyl)methyl, (2-tetrahydrothiofuranyl)methyl, and the like.

[0094] "Alkylamino" includes both monoalkylamino and dialkylamino, unless otherwise specified. "Monoalkylamino" refers to a group of (alkyl-NH)-; "dialkylamino" refers to a group of ((alkyl)2N)-. The alkyl group is defined in this document. The alkyl group is preferably a C1-C6 alkyl group. Examples include, but are not limited to, N-methylamino, N-ethylamino, N-isopropylamino, N,N-(diethyl)amino, and the like.

[0095] "Heteroalkylamino" refers to both mono-heteroalkylamino and di-heteroalkylamino, unless otherwise specified. Mono-heteroalkylamino refers to a group of (heteroalkyl-)NH-; di-heteroalkylamino refers to a group of (heteroalkyl)2N-. The definition of the "heteroalkyl" part is given in the relevant part of this article.

[0096] "Aminoalkyl" refers to a group of (amino-alkyl)-. The "alkyl" part is defined in this document. The aminoalkyl is preferably amino C1-C6 alkyl. It should be noted that "amino-C1-C6 alkyl" refers to a C1-C6 alkyl substituted by "amino", and its examples include but are not limited to: aminoethyl, 1-aminopropyl, 2-aminopropyl, etc.

[0097] "Arylamino" includes both mono-arylamino and di-arylamino, unless otherwise specified. Mono-arylamino refers to a group of (aryl-)NH-; di-arylamino refers to a group of formula (aryl)2N-; the definition of aryl is given in the relevant part of this article.

[0098] "Acyl" includes (alkyl-CO)-groups and (aryl-CO)-groups, unless otherwise specified. Wherein alkyl or aryl are as defined herein. Examples of acyl include, but are not limited to, acetyl, propionyl, isobutyryl, benzoyl, and the like.

[0099] "Acylamide" includes (alkyl-CONH)-groups and (aryl-CONH)-groups, unless otherwise specified. Wherein, alkyl or aryl are as defined herein. Examples of acylamide include, but are not limited to, acetylamide, propionylamide, butyramide, isobutyramide, benzylamide, and the like.

[0100] "Alkenyl" as a group or part of a group refers to an aliphatic hydrocarbon group containing at least one carbon-carbon double bond, which can be straight chain or branched. Alkenyl groups with C2-C14 are preferred. C2-C12 is more preferred; C2-C6 is most preferred. The group can contain multiple double bonds in its main chain and their conformation can be E or Z. Examples of alkenyl groups include, but are not limited to: vinyl, propenyl, etc.

[0101] "Alkynyl" as a group or part of a group refers to an aliphatic hydrocarbon group containing at least one carbon-carbon triple bond, which may be straight chain or branched. It is preferably C2-C14 alkynyl, more preferably C2-C12 alkynyl, and most preferably C2-C6 alkynyl. Examples of the alkynyl 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, etc.

[0102] "Alkoxy" refers to a group of (alkyl-O)-. The "alkyl" part thereof is defined in this document. The alkoxy is preferably a C1-C8 alkoxy, and more preferably a C1-C6 alkoxy. Examples of the alkoxy 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. can be cited.

[0103] "Alkenyloxy" refers to a group of (alkenyl-O)-, wherein alkenyl is as defined herein. C1-C6 alkenyloxy is preferred.

[0104] "Alkynyloxy" refers to a group of (alkynyl-O)-, wherein alkynyl is as defined herein. C1-C6 alkynyloxy is preferred.

[0105] "Alkoxycarbonyl" refers to a group of (alkyl-OC(O))-, wherein alkyl is as defined herein. The preferred alkyl group is C1-C6 alkyl. Examples thereof include, but are not limited to, methoxycarbonyl, ethoxycarbonyl, and the like.

[0106] "Alkylsulfinyl" refers to a group of (alkyl-S(O))-. Wherein, alkyl is as defined herein. The preferred alkyl is a C1-C6 alkyl group. Alkylsulfinyl groups include, but are not limited to, methylsulfinyl, ethylsulfinyl, and the like.

[0107] "Alkylsulfonyl" refers to a group of (alkyl-S(O)2-O)-, wherein alkyl is as defined herein. The preferred alkyl is a C1-C6 alkyl group. Examples include, but are not limited to, methylsulfonyl, ethylsulfonyl, and the like.

[0108] "Alkylaminocarbonyl" refers to an alkylamino-carbonyl group, wherein alkylamino is as defined herein.

[0109] "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, cycloheptylmethyl, and the like.

[0110] "Heterocycloalkenyl" refers to a heterocycloalkyl group containing at least one double bond. Heterocycloalkyl is defined herein.

[0111] "Aryl" as a group or part of a group refers to: (1) an aromatic monocyclic or condensed ring; preferably an aromatic carbocyclic ring (a ring structure in which all the ring atoms are carbon) having 5-12 carbon atoms. Examples of aryl include, but are not limited to, phenyl and naphthyl; (2) partially saturated carbocyclic rings may be connected, for example, phenyl and C5-7 cycloalkyl or C5-7 cycloalkenyl groups are fused to form a ring structure. Examples include, but are not limited to, tetrahydronaphthyl, indenyl or hydroindenyl. Aryl groups may be substituted by one or more substituents.

[0112] "Arylalkenyl" refers to a group of (aryl-alkenyl)-, wherein aryl and alkenyl are as defined herein. Exemplary arylalkenyl groups include, but are not limited to, phenylpropenyl, and the like.

[0113] "Aralkyl" refers to a group of (aryl-alkyl)-, wherein the aryl and alkyl parts are as defined herein. Exemplary aralkyl groups include, but are not limited to, benzyl, phenethyl, 1-naphthylmethyl, and the like.

[0114] "Cycloalkenyl" refers to a non-aromatic monocyclic or polycyclic ring system. It contains at least one carbon-carbon double bond and each ring preferably has 5-10 carbon atoms. Exemplary monocyclic cycloalkenyl rings include, but are not limited to: cyclopentene, cyclohexene or cycloheptene. The cycloalkenyl group may be substituted with one or more substituents.

[0115] "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 the aromatic group is a 4-15 membered heteroaryl group, more preferably a 5-7 membered heteroaryl group. Examples of the heteroaryl group include, for example, furanyl, 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.

[0116] "Heteroarylalkyl" refers to a group of (heteroaryl-alkyl)-, wherein the heteroaryl and alkyl parts are as defined herein. Exemplary heteroarylalkyl groups include, but are not limited to, 2-furylmethyl, 3-furylmethyl, 2-pyridylmethyl, and the like.

[0117] "Alkyl ether group" refers to a group of: (alkyl)-O-, wherein the "alkyl" part is as defined herein.

[0118] "Cycloalkyl ether group" refers to a group of: (cycloalkyl)-O-, wherein the "cycloalkyl" part is as defined herein.

[0119] "Heteroalkylether" refers to a group of: (heteroalkyl)-O-, wherein the "heteroalkyl" part is as defined herein.

[0120] "Aryl ether group" refers to a group of: (aryl)-O-, wherein the "aryl" part is as defined herein.

[0121] "Arylalkylether" refers to a group of: (aryl-alkyl)-O-, wherein the "aryl" and "alkyl" parts are as defined herein.

[0122] "Heteroaryl ether group" refers to a group of: (heteroaryl)-O-, wherein the "heteroaryl" part is as defined herein.

[0123] "Heteroarylalkylether" refers to a group of: (heteroaryl-alkyl)-O-, wherein the "heteroaryl" and "alkyl" parts are as defined herein.

[0124] "Heterocycloalkyl ether" refers to a group of: (heterocycloalkyl)-O-, wherein the "heterocycloalkyl" part is as defined herein.

[0125] "Heterocycloalkylamino" refers to both mono-heterocycloalkylamino and di-heterocycloalkylamino, unless otherwise specified. Mono-heterocycloalkylamino refers to a group of (heterocycloalkyl-)NH-; di-heterocycloalkylamino refers to a group of (heterocycloalkyl)2-N-. The definition of the "heterocycloalkyl" part is given in the relevant part of this article.

[0126] "Arylalkylamino" refers to both mono-arylalkylamino and di-arylalkylamino, unless otherwise specified. Mono-arylalkylamino refers to a group of (aryl-alkyl)-NH-; di-arylalkylamino refers to a group of (aryl-alkyl)2-N-. The definitions of the "aryl" and "alkyl" parts are as described in the relevant part of this article.

[0127] "Cycloalkylamino" refers to both mono-cycloalkylamino and di-cycloalkylamino, unless otherwise specified. Mono-cycloalkylamino refers to a group of (cycloalkyl)-NH-; di-arylalkylamino refers to a group of (cycloalkyl)2-N-. The definition of the "cycloalkyl" part is given in the relevant part of this article.

[0128] "Arylamino" refers to both mono-arylamino and di-arylamino, unless otherwise specified. Mono-arylamino refers to a group of (aryl)-NH-; di-arylamino refers to a group of (aryl)2-N-. The definition of the "aryl" part is given in the relevant part of this article.

[0129] "Heteroarylamino" refers to both mono-heteroarylamino and di-heteroarylamino, unless otherwise specified. Mono-heteroarylamino refers to a group of (heteroaryl)-NH-; di-heteroarylamino refers to a group of (heteroaryl)2-N-. The definition of the "heteroaryl" part is given in the relevant part of this article.

[0130] "Heteroarylalkylamino" refers to both mono-heteroarylalkylamino and di-heteroarylalkylamino, unless otherwise specified. Mono-heteroarylalkylamino refers to a group of (heteroaryl-alkyl)-NH-; di-heteroarylalkylamino refers to a group of (heteroaryl-alkyl)2-N-. The definitions of the "heteroaryl" and "alkyl" parts are as described in the relevant section of this document.

[0131] 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 of the hydrogen atoms is replaced by a valence; "heterocyclylene" refers to a heterocyclyl group in which one of the hydrogen atoms is replaced by a valence; "arylene" refers to an aryl group in which one of the hydrogen atoms is replaced by a valence; "alkylene" refers to an alkyl group in which one of the hydrogen atoms is replaced by a valence; "alkenylene" refers to an alkenyl group in which one of the hydrogen atoms is replaced by a valence; "cycloalkylene" refers to a cycloalkyl group in which one of the hydrogen atoms is replaced by a valence; "heteroarylene" refers to a heteroaryl group in which one of the hydrogen atoms is replaced by a valence; "heterocycloalkylene" refers to a heterocycloalkyl group in which one of the hydrogen atoms is replaced by a valence; "heterocycloalkenylene" refers to a heterocycloalkenyl group in which one of the hydrogen atoms is replaced by a valence; "alkyleneoxy" refers to an alkoxy group in which one of the hydrogen atoms is replaced by a valence; "alkenyleneoxy" refers to an alkenyloxy group in which one of the hydrogen atoms is replaced by a valence; "alkynyleneoxy" refers to an alkynyloxy group in which one of the hydrogen atoms 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.

[0132] The present invention includes compounds represented by the general formula (I) and possible various isomeric forms thereof, including non-mirror isomers, mirror isomers, tautomers, and geometric isomers of "E" or "Z" configuration isomers. Any chemist with a certain foundation can separate the above optically pure or stereoisomerically pure compounds.

[0133] 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.

[0134] In addition, the compounds represented by general formula (I) also include solvated and unsolvated forms of the compounds. Therefore, each formula includes compounds having the specified structure, including hydrated and unhydrated forms thereof.

[0135] In addition to the compounds represented by the general formula (I), kinase inhibitors of different embodiments include: pharmaceutically acceptable salts, prodrugs and active metabolites of such compounds and pharmaceutically acceptable salts of such metabolites.

[0136] The term "pharmaceutically acceptable salt" refers to certain salts of the above compounds that can maintain the original biological activity and are suitable for medical use. The pharmaceutically acceptable salts of the compounds represented by general formula (I) have two forms: one is a salt formed with an acid; the other is a salt formed with a base 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 acid organic 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 compounds represented by the general formula (I) include: lithium, sodium, potassium, magnesium, calcium, aluminum, zinc, etc.; bases that form pharmaceutically acceptable salts with the compounds represented by the general formula (I) include: choline, diethanolamine, morpholine, etc.

[0137] "Prodrug" is a derivative represented by general formula (I), which is converted into a compound represented by general formula (I) in vivo by means of in vivo metabolism (e.g., by hydrolysis, reduction or oxidation). For example, a compound represented by general formula (I) containing a hydroxyl group can be reacted with an acid to prepare a corresponding ester. The corresponding ester is a prodrug, which can be hydrolyzed into the parent drug in vivo. 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, etc.

[0138] 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, FGHR, PDGFR, VEGFR, TRK, EGFR, and B-RAF.

[0139] The administration of the compound represented by general formula (I) can be either enteral or parenteral. Enteral administration: oral or rectal. Parenteral administration includes: subcutaneous, intramuscular, intravenous and intradermal routes. Generally, the active compound represented by general formula (I) can be administered with a pharmaceutically acceptable carrier or diluent.

[0140] "Therapeutically effective amount" or "therapeutic amount" refers to an amount sufficient to produce a therapeutic effect. The effective amount can be administered in one or more doses. Generally, the effective amount is sufficient to alleviate, improve, stabilize, slow down or delay the further development of the disease.

[0141] The compounds of the present invention can be used alone, or in combination with one or more other drugs; or in combination with surgery or radiotherapy; or in combination with pharmaceutically acceptable carriers, diluents or excipients to form a certain dosage form for administration. The specific dosage form depends on the route of administration.

[0142] The pharmaceutical formulation for parenteral injection 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.

[0143] 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.

[0144] Solid dosage forms for oral administration include capsules, tablets, troches, powders and granules. In these solid dosage forms, the active compound represented by the 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 starches, 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.

[0145] Solid dosage forms of tablets, dragees, capsules, troches and granules can be prepared with coatings or shells.

[0146] The active compounds can also be administered in microencapsulated form, if desired, with one or more of the above-mentioned excipients.

[0147] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsifiers, 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, tetrahydrofuran alcohol, polyethylene glycol and fatty acid esters of sorbitan, etc.

[0148] Besides inert diluents, the oral compositions can also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

[0149] The suspension may contain, in addition to the active compound, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan esters.

[0150] Compositions for rectal or vaginal administration are preferably in the form of suppositories. These may be prepared by mixing the compounds of the invention with suitable non-irritating excipients or carriers.

[0151] 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 required preservatives, buffers or propellants.

[0152] The preferred dosage range is about 0.01-400 mg per kilogram of body weight per day. The more preferred dosage range is 0.2-100 mg per kilogram of body weight per day. An appropriate dosage may also be selected for multiple divided doses per day.

[0153] The compounds of the present invention can be used as, but not limited to, kinase inhibitors. The pyrimidine derivatives disclosed in the present invention can be used alone or together with other drugs or pharmaceutically acceptable carriers, diluents or excipients, and are suitable for preventing or treating diseases caused by, or associated with or accompanied by, the destruction of cell proliferation and / or angiogenesis. One example of these diseases is cancer.

[0154] The compounds of the present invention can also be used to treat diseases that involve or are at least partially regulated by RET, TRK, FGHR, PDGFR, and VEGFR activity, wherein RET and TRK activity are 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 and TRK inhibitors. Disorders of this type that are expected to be treated by the compounds of the invention include, but are not limited to, the following: antiproliferative disorders (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; ocular degenerative 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, Sjoegrens syndrome, multiple sclerosis, ankylosing spondylitis, membranous glomerulopathy, disc pain, systemic lupus erythematosus; diseases involving angiogenesis, including cancer, psoriasis, rheumatoid arthritis; psychological disorders, including bipolar disorder, 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, bacterial infections; viral infections such as: herpes simplex; protozoan infections such as: malaria, Leishmania infection, Trypanosoma brucei infection, toxoplasmosis and coccidiosis; hematopoietic disorders including: thalassemia, anemia and sickle cell anemia.

[0155] The term "cancer" as used herein generally refers to a broad range of diseases characterized by uncontrolled abnormal growth of cells.

[0156] The compounds of the present invention are expected to be useful in treating a variety of 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: adrenal cortical carcinoma, pancreatic cancer, pituitary cancer, thyroid cancer, parathyroid cancer, thymic carcinoma, multiple endocrine tumors; gastrointestinal cancers, including: : Gastric cancer, esophageal cancer, small intestine cancer, kidney cancer, renal cell carcinoma, liver 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 cancer, 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 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.

[0157] 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.

[0158] Preferred cancers that can be treated by the compounds of the invention are solid tumors and hematological malignancies.

[0159] In addition, the compounds of the present invention can be used to treat proliferative diseases that are resistant to other chemotherapeutic treatments; and to treat hyperproliferative diseases, such as leukemia, psoriasis, and the like.

[0160] Example_Synthesis and Application of Pyrimidine Derivatives

[0161] The compound represented by the general formula (I) can be synthesized using the synthetic route and method discussed below. The raw materials used are readily available. However, the synthetic route and method used in the present invention can be widely applied to the synthesis of analogs, and only the starting raw materials need to be changed. For example, the synthesis of compounds not described in detail in the examples herein can be synthesized by replacing the starting raw materials with the starting raw materials of the corresponding target compound, and slightly changing the reaction conditions when necessary according to common chemical knowledge to synthesize the desired target compound.

[0162] 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 know that the chemical reactions described can be applied to the preparation of various other compounds in different embodiments. For example: the synthesis of non-exemplified compounds can be successfully carried out by obvious modifications to those skilled in the art, or by changing to other appropriate reagents known in the art. For a list of suitable protecting groups in organic synthesis, see TW Greene's Protective Groups in Organic Synthesis, John Wiley & Sons, 1981. Other reactions disclosed herein or known in the art can be identified as having applicability for preparing other compounds of each embodiment.

[0163] Reagents useful for synthesizing the compounds can be obtained or prepared according to techniques known in the art.

[0164] In the following examples, all temperatures are in degrees Celsius unless otherwise indicated.

[0165] All starting materials and reagents were obtained from commercial suppliers, including but not limited to Aldrich Chemical Company, Lancaster Synthesis Ltd, etc. Commercially available starting materials and reagents were used directly without further purification unless otherwise specified.

[0166] The glassware was oven dried and / or heat dried. The reaction was followed on glass silica gel-60 F254 plates (0.25 mm) (TLC). Analytical thin layer chromatography was developed with appropriate solvent ratios (v / v). The end point of the reaction was when the starting material was exhausted on TLC.

[0167] Usually, the subsequent treatment is to double the volume of the reaction solution with the solvent used in the reaction, and then extract three times with 25% of the total volume of the extraction solvent, unless otherwise specified. The product-containing extract is dehydrated with anhydrous sodium sulfate, filtered on a rotary evaporator, and the solvent is evaporated under reduced pressure and attention is paid to the removal of the solvent in vacuum. Finally, the target compound is separated by flash column chromatography (J. Org. Chem., 1978; 43: 2923).

[0168] 1 H NMR spectra were measured using a Bruker instrument (400 MHz), and chemical shifts are expressed in ppm. Chloroform was used as a reference standard (7.25 ppm) or tetramethylsilane as an internal standard (0.00 ppm). Other solvents commonly used in NMR may also be used as needed. 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.

[0169] Mass spectra were obtained using LC / MS, with ESI or APCI ionization. All melting points are uncorrected.

[0170] The following examples are only used to illustrate the synthesis methods of the specific compounds invented. However, there is no limitation on the synthesis methods. Compounds not listed below can also be prepared by the same synthesis routes and methods as below, by selecting appropriate starting materials and slightly adjusting the reaction conditions according to common sense where necessary.

[0171] synthesis

[0172] The compound shown in the general formula (I), when Y = -NH, L 1 =-CH=CH-, L 2 =-C(O)-NH-, the synthesis method is as follows: a suitable substituted 6-chloro-2-aminopyrimidine derivative (II) is converted into the corresponding 6-chloro-2-iodopyrimidine derivative (III). The latter is condensed with a suitable amino derivative (I) to obtain compound (IV). Under amino protection, a Heck reaction is carried out with a suitable acrylate, and compound (IV) is converted into compound (VII) through intermediate (V). Alkaline hydrolysis is performed, and the resulting acid (VIII) is condensed with a suitable amino compound (IX) to obtain compound X. After hydrolysis to remove the protecting group, compound XI is obtained, i.e., the target compound shown in general formula-(I) (see synthetic route 1 for details).

[0173] Synthetic route 1

[0174]

[0175] In the general formula (I), when R1 is 5-methyl-1H-pyrazole, the synthesis of the corresponding R1-NH2 can be synthesized by the method shown in Synthesis Scheme 2: The nitro compound (XII) can be reduced under protection to obtain the corresponding amino compound I-1 (see Synthesis Scheme 1 for details)

[0176] Synthetic route 2

[0177]

[0178] The content of the present invention is further explained below with reference to examples. The purpose is to allow technicians with basic knowledge in the field to more clearly understand and practice the specific content of the present invention. However, the protection scope of the present invention is not limited to these examples.

[0179] Embodiment_1

[0180] Synthesis of (E,S)-N-(1-(6-(4-fluoro-1H-pyrazol-1-yl)pyridin-3-yl)ethyl)-3-(4-methyl-6-((5-methyl-1H-pyrazol-3-yl)amino)pyrimidin-2-yl)acrylamide (1).

[0181] Step-1 Synthesis of 4-chloro-2-iodo-6-methylpyrimidine (III-1):

[0182]

[0183] To a mixed solution of 4-chloro-6-methylpyrimidine-2-amine (II-1) (10.0 g, 69.4 mmol) and diiodomethane (55.8 g, 208.3 mmol) in acetonitrile (200 mL) was added dropwise tert-butyl nitrite (24.5 g, 208 mmol). Under nitrogen protection, the mixture was stirred at 60 ° C for 30 minutes and then at 80 ° C for 2 hours. The system was cooled to room temperature and concentrated. Water (100 mL) was added to dilute the residue and extracted with ethyl acetate (100 mL×2). The mixture was washed with Na2S2O3 aqueous solution (sat., 30 mL) and brine (30 mL). The organic phases were separated and combined, dried over Na2SO4, and concentrated to obtain a crude product, which was purified by silica gel column chromatography (petroleum ether: ethyl acetate from 100:1 to 10:1) to obtain the target compound III-1 (8.82 g, yield 50%), purity: 95% (254 nm); 1 H NMR(400MHz, CDCl3): δ7.20(s,1H),2.49(s,3H).ESI-MS(m / z):254.9[M+H] + .

[0184] Step-2 Synthesis of 2-iodo-6-methyl-N-(5-methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-3-yl)pyrimidin-4-amine (IV-1):

[0185]

[0186] DIEA (13.2 g, 103 mmol) was added to a solution of 4-chloro-2-iodo-6-methylpyrimidine (III-1) (8.70 g, 34.2 mmol) and 5-methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-3-amine (3) (I-1, 7.42 g, 41.0 mmol) in anhydrous ethanol (200 mL), and the mixture was refluxed for 2 days. The mixture was cooled and concentrated, and water (50 mL) and ethyl acetate (50 mL) were added to dilute the residue. The mixture was extracted with ethyl acetate (50 mL×3), and the combined organic phase was washed with brine (20 mL), dried over Na2SO4, and concentrated to obtain a crude product, which was purified by silica gel column chromatography (petroleum ether: ethyl acetate from 10:1 to 3:1) to obtain the target compound IV-1 (6.82 g, yield 50%) as a yellow solid. The product purity was 73%, which was directly used in the next step. 1 H NMR (400MHz, DMSO-d6) δ10.18(s,1H),6.16(s,1H),5.31(dd,J=4,0.8Hz,1H),3.84-3.93(m,1H),3.59-3.66(m,1H),2.28( s,3H),2.23(s,3H),1.97-2.01(m,2H),1.83-1.88(m,1H),1.65-1.69(m,1H),1.52-1.55(m,2H).ESI-MS(m / z):400.0[M+H] + .

[0187] Step-3 Synthesis of tert-butyl (2-iodo-6-methylpyrimidin-4-yl) (5-methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-3-yl) carbamate (V-1)

[0188]

[0189] To a solution of 2-iodo-6-methyl-N-(5-methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-3-yl)pyrimidine-4-amine (6.80 g, 17.0 mmol) and Boc2O (4.46 g, 20.5 mmol) in acetonitrile (100 mL) were added triethylamine (3.44 g, 34.1 mmol) and DMAP (50 mg), the temperature was raised to 80°C and stirred for 1 hour, the mixture was cooled and concentrated, water (50 mL) and ethyl acetate (50 mL) were added to dilute the residue, and the mixture was extracted three times with ethyl acetate (50 mL×3), the organic phases were combined, washed with brine (20 mL), dried over Na2SO4, and concentrated to give a crude product, which was purified by silica gel column chromatography (petroleum ether:ethyl acetate from 10:1 to 8:1) to give the target compound V-1 (4.55 g, yield 54%) as a light yellow oil. 1 H NMR (400MHz, CDCl3): δ7.52(s,1H),5.99(s,1H),5.33(dd,J=8.0,2.8Hz,1H),3.95-3.92(m,1H),3.66-3.61(m,1H),2.43(s,3H),2.37(s,3 H),2.32-2.25(m,1H),2.12-2.07(m,1H),1.93-1.88(m,1H),1.70-1.65(m,2H),1.59-1.57(m,1H),1.46(s,9H).ESI-MS(m / z):499.9[M+H] + .

[0190] Step-4 Synthesis of (E)-ethyl 3-(4-((tert-butyloxycarbonyl)(5-methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-3-yl)amino)-6-methylpyrimidin-2-yl)acrylate (VII-1):

[0191]

[0192] To a mixture of tert-butyl (2-iodo-6-methylpyrimidin-4-yl)(5-methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-3-yl)carbamate (V-1) (1.0 g, 2.0 mmol), ethyl acrylate VI-1 (600 mg, 6.0 mmol), TEA (1.01 g, 10.0 mmol) in CH3CN (20 mL) was added Pd(OAc)2 (135 mg, 0.6 mmol), and the temperature was raised to 100°C under nitrogen protection and stirred for 5 hours. After cooling, the reaction mixture was concentrated to give a crude product, which was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 6:1 to 3:1) to give the desired compound VII-1 (330 mg, yield 49%) as a brown oil.1 H NMR (400MHz, CDCl3): δ7.58(s,1H),7.48(d,J=16.0Hz,1H),6.85(d,J=15.6Hz,1H),6 .00(s,1H),5.33-5.30(m,1H),4.23(q,J=6.8Hz,2H),3.98-3.95(m,1H),3.66-3.61(m ,1H),2.51(s,3H),2.39(s,3H),2.32-2.29(m,1H),2.09-2.05(m,1H),1.92-1.88(m, 1H),1.67-1.55(m,3H),1.46(s,9H),1.30(t,J=6.8Hz,3H).ESI-MS(m / z):472.4[M+H] +

[0193] Step-5 Synthesis of (E)-3-(4-((tert-butyloxycarbonyl)(5-methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-3-yl)amino)-6-methylpyrimidin-2-yl)acrylic acid (VIII-1):

[0194]

[0195] To a solution of (E)-ethyl 3-(4-((tert-butoxycarbonyl)(5-methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-3-yl)amino)-6-methylpyrimidin-2-yl)acrylate (VII-1) (350 mg, 0.74 mmol) in methanol (2 mL) was added LiOH aqueous solution (LiOH·H2O (47 mg, 1.11 mmol), water (1 mL)) at room temperature, and the mixture was stirred at 30°C for 1 hour. The reaction mixture was concentrated, and the residue was diluted with water (20 mL) and extracted with ethyl acetate (20 mL). The aqueous phase was separated, acidified to pH = 4 with 10% HCl aqueous solution, and then extracted with ethyl acetate (15 mL x 4). The combined organic phases were washed with brine (10 mL), dried over Na2SO4, and concentrated to give VIII-1 as a yellow solid (250 mg, yield 76%). 1H NMR (400MHz, DMSO-d6) δ12.80(s,1H),7.43(s,1H),7.21(d,J=16Hz,1H),6.75(d,J=16H z,1H),6.10(d,J=0.8Hz,1H),5.41(dd,J=4.0,2.8Hz,1H),3.83(d,J=12Hz,1H),3.61(dd d,J=4Hz,1H),2.49(s,3H),2.33(s,3H),2.06-2.15(m,1H),1.91-1.95(m,1H),1.76-1. 81(m,1H),1.61-1.67(m,1H),1.47-1.52(m,2H),1.42(s,9H).ESI-MS(m / z):444.4[M+H] + .

[0196] Step-6 Synthesis of (E, S)-tert-butyl (2-(3-((1-(6-(4-fluoro-1H-pyrazol-1-yl)pyridin-3-yl)ethyl)amino)-3-oxapropyl-1-en-1-yl)-6-methylpyrimidin-4-yl)(5-methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-3-yl)carbamate (X-1):

[0197]

[0198] Under ice bath condition, to a mixture of (E)-3-(4-((tert-butyloxycarbonyl)(5-methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-3-yl)amino)-6-methylpyrimidin-2-yl)acrylic acid (VIII-1) (600 mg, 1.35 mmol) and DIEA (349.7 mg, 2.71 mmol) in DCM (18 mL) was added HATU (617.3 mg, 1.62 mmol). After stirring for 30 minutes, (S)-1-(6-(4-fluoro-1H-pyrazol-1-yl)pyridin-3-yl)ethanamine (IX-1) (306.7 mg, 1.49 mmol) was added. The reaction was stirred at room temperature for 12 hours. The reaction was complete by TLC. The system was concentrated, the residue was diluted with water (15 mL), and extracted with ethyl acetate (40 mL x 2). The organic phases were separated and combined, and the mixture was washed with brine (40 mL x 2). 5) Wash, dry with Na2SO4 and concentrate to obtain a crude product, which was chromatographed on a large thin layer plate (PE:EA=1:1) to obtain compound X-1 (600 mg, yield 70%) as a white solid. ESI-MS (m / z): 632.4 [M+H] + .

[0199] Step-7 Synthesis of (E,S)-N-(1-(6-(4-fluoro-1H-pyrazol-1-yl)pyridin-3-yl)ethyl)-3-(4-methyl-6-((5-methyl-1H-pyrazol-3-yl)amino)pyrimidin-2-yl)acrylamide 1(XI-1).

[0200]

[0201] To a solution of (E,R)-tert-butyl(2-(3-(1-(6-(4-fluoro-1H-pyrazol-1-yl)pyridin-3-yl)amino)-3-oxopropin-1-ene-1-yl)-6-methylpyrimidin-4-yl)(5-methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-3-yl)carbamate (600 mg, 0.95 mmol) in DCM (15 mL) was added TFA (5 mL) at room temperature. After stirring at room temperature for 4 hours, the reaction was complete by TLC. The product was concentrated in vacuo to give a residue, which was diluted with ethyl acetate (80 mL) and neutralized by adding aqueous NaHCO3 solution (50 mL). The organic phase was separated and the combined organic phases were washed with brine (40 mL), dried over Na2SO4, filtered and the filtrate was recrystallized to give yellow solid Compound 1 (300 mg, yield 71%). 1 H NMR (400 MHz, DMSO-d6): 1 H NMR (400MHz, DMSO-d6) δ12.01(s,1H),9.81(s,1H),9.00(d,J=4Hz,1H),8.68(d,J=4Hz,1H),8.45(d,J=4Hz,1H),7.87-8.01(m,3H),7.1 5-7.27(m,2H),6.97(s,1H),6.19(s,1H),5.13(p,J=12Hz,1H),2.32(s,3H),2.24(s,3H),1.48(d,J=8Hz,3H).ESI-MS(m / z):448.3[M+H] + .

[0202] Example 2-100

[0203] According to the method of Example 1, a wide variety of derivatives can be synthesized by simply changing the appropriate starting materials. Examples 2-100 are some representative examples (see Table 1).

[0204] Table 1 Representative pyrimidine derivatives of general formula (I)

[0205]

[0206]

[0207]

[0208]

[0209]

[0210]

[0211]

[0212]

[0213]

[0214]

[0215]

[0216]

[0217]

[0218]

[0219]

[0220]

[0221]

[0222]

[0223]

[0224] In addition, referring to the method of Example 1, as long as the starting raw materials are appropriately selected, a wider variety of derivatives can be synthesized, such as the compounds listed in Table 2.

[0225] Table 2 Other derivatives included in the general formula (I)

[0226]

[0227]

[0228]

[0229]

[0230]

[0231]

[0232] Biological experiments and pharmacodynamic analysis

[0233] Example 167 Detection of the inhibitory activity of the compounds of the present invention on related kinases

[0234] There are many literature reports on the test of kinase activity, and there are also related kinase detection kits available. Products from Cisbio can be selected but are not limited to: HTRF kinase-STK KIT. Taking the detection of RET kinase inhibitory activity by HTRF (homogeneous time-resolved fluorescence) kinase detection kit as an example, the experimental operation steps are as follows.

[0235] 1. Experimental methods and steps:

[0236] 1.1 Prepare the test compound into a 10 mM (mmol / L) DMSO solution.

[0237] 1.2 The 10mM 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 diluted to 9 concentrations in a 3-fold gradient, and the concentrations were: 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 to 2.5μM (2.5× compound) working solution, and then 2.5μM was used as the highest concentration, and the working solution was diluted to 9 concentrations in a 3-fold gradient ... The buffer was diluted to a 2.5 μM working solution, and then 2.5 μM was used as the highest concentration and diluted in a 3-fold gradient to continuously dilute the 2.5× working solution of the test compound to 9 concentrations, namely 2.5, 0.833333, 0.277778, 0.092593, 0.030864, ​​0.010288, 0.003429, 0.001143, and 0.000381 μM.

[0238] 1.3 Take 4 μL of 2.5× compound working solution and add it to a 384-well plate (Greiner, Cat#781280), set up Blank wells (no compound and kinase) and Control wells (only kinase without compound), and add 4 μL kinase buffer to the Blank and Control wells.

[0239] 1.4 Prepare the kinase storage solution into the corresponding 5× working solution with kinase buffer, take 2μL of 5× kinase working solution and add it to each well containing compound working solution, add 2μL kinase buffer to the Blank well, and add 2μL 5× kinase working solution to the Control well.

[0240] 1.5 Take 2 μL of 5× substrate storage solution (TK Antibody-Cryptate) and add it to each well containing compound and kinase working solution, add 2 μL kinase buffer to the Blank well, and add 2 μL 5× substrate storage solution to the Control well.

[0241] 1.6 Take 2 μL of ATP working solution (5×) and add it to each detection well.

[0242] 1.7 Cover the 384-well plate with a sealing film and incubate at 37°C for 1 hour, then add 5 μL (4X) of reaction stop solution (Streptavidin-XL665) to each well;

[0243] 1.8 Continue to seal the 384-well plate, incubate at 37°C for 1 hour, and then detect the 665 and 620 signal values ​​on the 2104 EnVision plate reader.

[0244] Preparation of detection system working solution:

[0245]

[0246]

[0247] 2. Data Analysis:

[0248] The inhibition rate (IR) of the test compound was calculated using the following formula: IR (%) = (RLUCTR (665 / 620) – RLU compound (665 / 620)) / (RLU CTR (665 / 620) – RLU BLANK (665 / 620)) * 100%. The inhibition rates of different concentrations of the compound were calculated in Excel, and then the IC was calculated using GraphPad Prism 5 software. 50 .

[0249] 3. Activity inhibition test results of wild-type RET and mutant RET kinases

[0250] Some biological activity test results are shown in Table 3

[0251] Table 3 Kinase inhibitory activity of target compounds

[0252]

[0253] Similarly, other compounds were also tested to inhibit the activity of wild-type RET and mutant RET kinases, showing good inhibitory effects.

[0254] At the same time, the compounds of the present invention have obvious inhibitory effects on other kinases, such as TRK, RAF, FGHR, PDGFR, VEGFR and other kinases.

[0255] Example 168 Detection of the cell inhibitory activity of the compounds of the present invention

[0256] The cell activity test uses CTG (CELL TITER-GLO) luminescence method to test the activity of the target compound. The principle is: ATP adenine nucleoside triphosphate (ATP for short) 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 are used to measure the luminescence value. 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-proliferation activity data of the compound on tumor cells can be obtained according to the calculation formula.

[0257] 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 effect on human thyroid ductal carcinoma cells (TT) and human colon cancer cells (KM12); in order to determine its activity on RET Fusion, the growth inhibitory activity of the target compound on KIF5B-RET fusion cells was specifically tested.

[0258] 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).

[0259] 1. Determination of the activity of compounds on human thyroid ductal carcinoma cells and human colon cancer cells

[0260] The following method is used to determine the effect of compounds on tumor cell proliferation by using the CTG luminescence method.

[0261] Specific experimental operation methods and processes:

[0262] 1.1 Cell recovery

[0263] Immediately place the frozen cells from the liquid nitrogen storage tank in a 37°C constant temperature water bath and shake for 2 minutes. After the cell freezing solution is completely thawed, transfer the cell suspension into a 15mL centrifuge tube, slowly add 4mL of culture solution, centrifuge (1000r / min, 5min), discard the supernatant, aspirate the original solution, add 5mL of the above culture medium, gently blow to a single cell suspension, transfer it to a culture bottle, and place it in an incubator for culture.

[0264] 1.2 Cell culture

[0265] The cells were cultured with complete medium in an incubator at 37°C and 5% CO 2 . The cells were passaged regularly and cells in the logarithmic growth phase were used for plating.

[0266] 1.3 Cell plating

[0267] The cells were stained with trypan blue and the living cells were counted. The cell concentration was adjusted to the appropriate medium plate concentration (TT: 50000 cell / mL, KM12: 35000 cell / mL). 90 μL of cell suspension was added to each well of a 96-well culture plate (Corning, 3599), and blank control wells and solvent control wells were set. The culture medium containing cells was added to the blank control wells, and the culture medium without cells was added to the solvent control wells. The culture plate was then placed in an incubator at 37°C, 5% CO2, and 100% relative humidity for overnight culture.

[0268] 1.4 Compound preparation

[0269] Weigh the compound and prepare a 10mM stock solution with DMSO. Dilute the stock solution of the compound to be tested with serum-free medium to a final concentration of 100μM 10× compound working solution (including reference substance) on a dispensing plate (Beaver, Suzhou). Dilute with serum-free medium in a 3-fold concentration gradient to obtain 9 concentration gradients of 10× compound working solutions, with compound concentrations of 100, 33.33, 11.11, 3.70, 1.23, 0.411, 0.137, 0.046, and 0.015μM, respectively.

[0270] 1.5 Addition of compounds

[0271] Add 10× compound working solution of different concentration gradients to the 96-well cell culture plate, 10 μL / well, add 10 μL DMSO-cell culture solution mixture to the solvent control well and blank control well, the final DMSO concentration is 0.1%, and set 2 replicates for each concentration. Return the 96-well cell plate to the 37°C, 5% carbon dioxide incubator for 5 days.

[0272] 1.6CTG detection

[0273] The cell culture plate was taken out and placed for 30 minutes to equilibrate to room temperature, 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, and 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.

[0274] 1.7 Data Analysis

[0275] The inhibition rate (IR) of the test compound was calculated using the following formula: IR (%) = (1-(RLU compound-RLU blank control) / (RLU solvent control-RLU blank control))*100%. Finally, nonlinear regression analysis was performed using the logarithm of compound concentration-inhibition rate in Graphpadprism 5 software to obtain the IC value of the compound for inhibiting cell proliferation. 50 value.

[0276] Results The test results are shown in Table 4.

[0277] Table 4 Results of the inhibitory activity test on TT cells and KM12 cells obtained by the compounds

[0278]

[0279]

[0280]

[0281]

[0282] 2. Determination of compound activity against RET fusion engineered cell lines

[0283] 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 the target compounds against KIF5B-RET Fusion and CCDC6-RETFusion.

[0284] The following method is used to determine the effect of the compound on the proliferation of KIF5B-RET and CCDC6-RET fusion engineered cells, which is determined by using the CTG luminescence method. For the fusion of KIF5B-RET and CCDC6-RET, Ba / F3-KIF5B-RET and Ba / F3-CCDC6-RET fusion engineered cell lines (constructed by Precedo) were used and cultured according to the corresponding conditions.

[0285] 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).

[0286] The experimental operation method is as follows

[0287] 2.1 Cell recovery

[0288] Immediately place the frozen cells from the liquid nitrogen storage tank in a 37°C constant temperature water bath and shake for 2 minutes. After the cell freezing solution is completely thawed, transfer the cell suspension into a 15mL centrifuge tube, slowly add 4mL of culture solution, centrifuge (1000r / min, 5min), discard the supernatant, aspirate the original solution, add 5mL of the above culture medium, gently blow to a single cell suspension, transfer it to a culture bottle, and place it in an incubator for culture.

[0289] 2.2 Cell culture

[0290] Ba / F3-KIF5B-RET and Ba / F3-CCDC6-RET engineered cells were cultured in complete medium (RPMI-1640+10% FBS+1% P / S) in an incubator at 37°C and 5% CO2. Cells were passaged regularly and cells in the logarithmic growth phase were used for plating.

[0291] 2.3 Cell plating

[0292] The cells were stained with trypan blue and the living cells were counted. The cell concentration was adjusted to 30,000 cells / mL of cell solution. 90 μL of cell suspension was added to each well of a 96-well culture plate (Corning, 3599), and blank control wells and solvent control wells were set. The culture solution containing cells was added to the blank control wells, and the culture solution without cells was added to the solvent control wells. The culture plate was then placed in an incubator at 37°C, 5% CO2, and 100% relative humidity for overnight culture.

[0293] 2.4 Compound preparation

[0294] Weigh the compound and prepare a 10mM stock solution with DMSO. Dilute the stock solution of the compound to be tested with serum-free medium to a final concentration of 100μM 10× compound working solution (including reference substance) on a dispensing plate (Beaver, Suzhou). Dilute with serum-free medium in a 3-fold concentration gradient to obtain 9 concentration gradients of 10× compound working solutions, with compound concentrations of 100, 33.33, 11.11, 3.70, 1.23, 0.411, 0.137, 0.046, and 0.015μM, respectively.

[0295] 2.5 Addition of compounds

[0296] Add 10× compound working solution of different concentration gradients to the 96-well cell culture plate, 10 μL / well, add 10 μL DMSO-cell culture solution mixture to the solvent control well and blank control well, the final DMSO concentration is 0.1%, and set 2 replicates for each concentration. Return the 96-well cell plate to the 37°C, 5% carbon dioxide incubator for 5 days.

[0297] 2.6CTG detection

[0298] The cell culture plate was taken out and placed for 30 minutes to equilibrate to room temperature, 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, and 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.

[0299] 2.7 Data Analysis

[0300] The inhibition rate (IR) of the test compound was calculated using the following formula: IR (%) = (1-(RLU compound-RLU blank control) / (RLU solvent control-RLU blank control))*100%. Finally, nonlinear regression analysis was performed using the logarithm of compound concentration-inhibition rate in Graphpadprism 5 software to obtain the IC value of the compound for inhibiting cell proliferation. 50 value.

[0301] The test results are shown in Table 5.

[0302] Table 5. Results of the assay of the inhibitory activity of the obtained compounds on the growth of RET fusion engineered cells

[0303]

[0304] The results showed that the target compound had significant inhibitory activity against KIF5B-RET Fusion and CCDC6-RET Fusion. Data for the remaining compounds were not provided, but they all had significant inhibitory effects.

[0305] Example 169 In vivo antitumor activity of the compounds of the present invention

[0306] Some compounds with strong in vitro activity and low toxicity were selected to determine the maximum tolerated dose (MTD) in mice. The anti-tumor activity of the compounds of the present invention in vivo was determined on a human cancer nude mouse xenograft tumor model to explore the dosage, route of administration, frequency and cycle of administration of the test compounds to produce pharmacodynamic effects.

[0307] Female BALB / C nude mice aged 5-6 weeks, weighing approximately 18-20 g, were raised.

[0308] The efficacy of the subcutaneous cell transplanted tumors was studied and evaluated in the BALB / c nude mouse model.

[0309] Construction of human cancer nude mouse xenograft tumor model: TT (human thyroid cancer cell) culture, monolayer cultured tumor cells were digested and detached from the wall, collected and resuspended in serum-free culture medium, and adjusted to a concentration of 5×10 6 / 0.2mL, put it in an ice box and bring it to the animal room. Use a syringe with a No. 6 needle to directly take 0.2mL of cell suspension and transplant it subcutaneously in the scapula behind the left armpit of nude mice. 5×10 6 / 0.2mL / mouse, measure the tumor volume every 2-3 days, select nude mice with vigorous tumor growth and no ulceration, remove the tumor under sterile conditions, cut the tumor tissue into pieces with a diameter of about 2-3mm, and inoculate it subcutaneously at the scapula behind the left armpit of the nude mouse. After three generations, when the tumor volume grows to 100-150mm 3 Nude mice with tumors that were too large or too small were removed and randomly divided into groups for drug administration.

[0310] Randomly divided into 5 groups, including negative control group (solvent), positive control group (Blu-667, 10 mg / kg), high, medium and low dose treatment groups (5 mg / kg, 10 mg / kg, 40 mg / kg, respectively, of which the high dose is lower than MTD), 5 nude mice in each group, intraperitoneal injection, twice a day, for 3 consecutive weeks. During this period, the animal weight, tumor volume and animal death were detected every 3 days. The animals were killed 24 hours after the last administration, and the tumor volume, tumor weight and nude mouse weight were measured. The tumor volume growth curve, nude mouse weight growth curve and tumor inhibition rate, animal mortality rate were drawn, and the relative tumor proliferation rate T / C (%) was calculated according to the formula T / C (%) = TRTV / CRTV*100%. (TRTV: treatment group RTV; CRTV: negative control group RTV, relative tumor volume RTV = Vt / V0, where V0 is the tumor volume at the time of group administration, and Vt is the tumor volume after administration). The in vivo anti-tumor efficacy of the compounds of the present invention is that the relative tumor proliferation rate T / C (%) is ≤40%, and the TGI% is greater than 80% (see Figure 1The anti-tumor inhibitory activity of compound 1 in the TT model was statistically significant, with obvious pharmacodynamic effects and good in vivo tumor inhibition effects. At the same time, the body weight of animals in different dose groups of compound 1 of the present invention did not decrease significantly, showing good tolerance. Similarly, other compounds also have similar in vivo tumor inhibition effects.

[0311] In this experiment, compound 1 was administered by oral gavage to test its in vivo efficacy in the TT xenograft tumor model, while Blu-667 was used as a control drug. The test results showed that compound 1 had a T / C ratio of less than 40% and a TGI% of more than 80% at doses of 5, 10, and 40 mg / kg, showing a good in vivo tumor inhibition effect. At the same time, there was no significant decrease in the body weight of animals in different dose groups of compound 1, showing good tolerability.

[0312] In addition to using relevant tumor cell lines, relevant engineered strains, such as KIF5B-RET fusion tumor cell lines, should also be used for in vivo anti-tumor inhibitory activity.

[0313] The details of the specific examples described in the present invention are not to be construed as limitations thereof. Various synonyms and modifications may be made without departing from the essence and scope of the present invention, and it is known that these synonymous embodiments are part of the present invention.

Claims

1. A pyrimidine derivative, characterized in that: The pyrimidine derivatives are compounds represented by the general formula (I), and their optical isomers or pharmaceutically acceptable salts thereof: In the above general formula (I), R 1 Selected from: R 4 Selected from C1-C3 alkyl, C3-C8 cycloalkyl; Y is: -NH-; R 2 Selected from: hydrogen atom, alkyl; any of the above groups is independently unsubstituted or substituted by one or more substituents, and these substituents are halogen; the alkyl is a C1-C8 straight chain or branched alkyl; L 1 Selected from: -CH=CH-, -C≡C-; L 2 Selected from: carbonyl, -CO-NH-; R 3 Selected from Benzene ring, substituted benzene ring, substituted pyridine, pyrimidine, substituted pyrimidine; The substituted benzene ring contains 1-3 substituents, and the substituents are selected from halogen, C1-C3 alkyl, methyl ether, trifluoromethyl ether, ethyl ether, trifluoroethyl ether, propyl ether, trifluoropropyl ether, trifluoromethyl, amino, The substituted pyridine contains 1-3 substituents, and the substituents are selected from halogen, C1-C3 alkyl, methyl ether, trifluoromethyl ether, ethyl ether, trifluoroethyl ether, propyl ether, trifluoropropyl ether, trifluoromethyl, The substituted pyrimidine contains 1 to 3 substituents selected from Amino.

2. The pyrimidine derivative according to claim 1, characterized in that: R 2 It is a C1-C5 straight chain or branched chain alkyl.

3. A pyrimidine derivative, characterized in that: Its structure is selected from one of the following structures, or its optical isomers or pharmaceutically acceptable salts:

4. The method for preparing a pyrimidine derivative according to any one of claims 1 to 3, characterized in that: When Y=NH, L1=CH=CH, L2=CO-NH, the preparation method includes: S1, converting the substituted 6-chloro-2-aminopyrimidine derivative II into the corresponding 6-chloro-2-iodopyrimidine derivative III; S2, 6-chloro-2-iodopyrimidine derivative III is condensed with amino derivative I to obtain compound IV; S3, under amino protection, compound IV generates intermediate V, which undergoes Heck reaction with acrylate to be converted into compound VII; S4, alkaline hydrolysis of compound VII, condensation of the obtained acid VIII with the amino compound IX to obtain compound X; S5. Compound X is hydrolyzed to remove the protecting group to obtain compound XI, i.e., the target compound represented by general formula (I); The structures of the compounds involved include:

5. Any pharmaceutical dosage form comprising the pyrimidine derivative according to any one of claims 1 to 3 and a pharmaceutically acceptable diluent, excipient or carrier.

6. Use of the pyrimidine derivative according to any one of claims 1 to 3 in the preparation of a drug for treating cancer.

7. The use according to claim 6, characterized in that The cancer is selected from the group consisting of bone cancer, endocrine cancer, gastrointestinal cancer, genitourinary cancer, head and neck tumors, blood cancer, lung cancer, lymphoma, eye cancer, skin cancer, and soft tissue sarcoma.

8. The use according to claim 6, characterized in that The cancer is selected from the group consisting of: Ewing's sarcoma, osteosarcoma, acoustic neuroma, neuroblastoma, glioma, spinal cord tumor, breast cancer, colorectal cancer, adrenocortical carcinoma, pancreatic cancer, pituitary cancer, thyroid cancer, parathyroid cancer, thymic cancer, gastric cancer, esophageal cancer, small intestine cancer, kidney cancer, liver cancer, extrahepatic bile duct cancer, gallbladder cancer, testicular cancer, penile cancer, prostate cancer, cervical cancer, ovarian cancer, vaginal cancer, endometrial cancer, gestational trophoblastic tumor, fallopian tube cancer, uterine sarcoma, cancer, oral cancer, lip cancer, salivary gland cancer, laryngeal cancer, paranasal sinus cancer, nasopharyngeal cancer, myelodysplastic syndrome, myeloproliferative disorders, aplastic anemia, Fanvonis anemia, essential macroglobulinemia, small cell lung cancer, non-small cell lung cancer, Hodgkin's disease, non-Hodgkin's lymphoma, retinoblastoma, melanoma, non-melanoma skin cancer, Merkel cell carcinoma, childhood soft tissue sarcoma, adult soft tissue sarcoma, Kaposi's sarcoma, bladder cancer, and urethral cancer.

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