CN106749223B - Tyrosine kinase inhibitor and preparation method and application thereof - Google Patents

Abstract

The invention relates to a tyrosine kinase inhibitor and a preparation method and application thereof, belonging to the technical field of pharmaceutical chemistry. The tyrosine kinase inhibitor with the structural characteristics of the general formula I or pharmaceutically acceptable salt or stereoisomer thereof can effectively inhibit the activity of tyrosine kinase. The compound has an inhibitory effect on kinases such as DDR1, DDR2, Abl, Src, Btk, Kit and the like, the half inhibitory concentration of the compound is compared with or superior to that of a positive control dasatinib, or at least equivalent to that of the positive control, particularly the compound 8j has a half inhibitory concentration which is less than that of the positive control on five tyrosine kinases such as DDR1, DDR2, Src, Btk and Kit, and K562 cells, has good enzyme inhibitory activity and cell activity, and has a wide application prospect.

Description

Tyrosine kinase inhibitor and preparation method and application thereof

Technical Field

The invention relates to the technical field of medicinal chemistry, in particular to a tyrosine kinase inhibitor and a preparation method and application thereof.

Background

Protein kinases are one of the largest families that make up human enzymes, and regulate many different signaling processes by adding phosphate groups to proteins. In particular, tyrosine kinases phosphorylate proteins on the phenol portion of tyrosine residues. The tyrosine kinase family includes members that control cell growth, migration, and differentiation. Aberrant kinase activity has been manifested in a number of human diseases, including cancer, autoimmune disorders, and inflammatory diseases, among others. Since protein kinases belong to key regulators of cell signaling, they are targets for the regulation of cellular function with small molecule kinase inhibitors and are therefore good drug design targets.

Chronic Myelogenous Leukemia (CML) is a malignant tumor of the hematopoietic system with a very high mortality rate. The natural pathogenesis of CML can be roughly divided into three stages: chronic Phase (CP), Accelerated Phase (AP) and acute Phase (BP). From the initial stage to the acute stage, the number of abnormal cells in the bone marrow rises and spreads to the outside of the bone marrow. The chronic phase can typically be latent for 4 to 5 years, during which time the patient is less symptomatic and can only be detected by blood tests. Most CML patients in the chronic phase can be treated with Chemotherapy (Chemotherapy), Interferon (Interferon) therapy or Bone Marrow Transplantation (Bone Marrow Transplantation). After the progression of CML from the chronic phase to the accelerated phase, the expansion of leukocytes in patients is often refractory to conventional therapy. The accelerated phase is usually maintained between 6 and 18 months, followed by a catastrophe phase, in which the patient can only survive for 1 to 6 months.

Clinical diagnosis shows that chronic myelogenous leukemia (CML, lentil for short) is mainly produced by chromosomal abnormalities. More than 95% of CML patients can detect the Ph chromosome, i.e., t (9; 22) (q 34; ql). The Abl gene located on chromosome 9 is translocated to the Bcr gene on chromosome 22 to form a Bcr/Abl fusion gene, and the chromosome in which the fusion gene is located is named as the Ph chromosome. The tyrosine kinase protein expressed by the Bcr/Abl fusion gene has uncontrollable and continuous physiological activity, so that a Signal transmission pathway (Signal path) for regulating cell replication is always in an active state, more abnormal white blood cells are continuously generated, and the generation of white blood cells normally controlled by bone marrow is destroyed, thereby forming the CML disease.

Imatinib (Imatinib, in vitro Activity, IC of K562, a drug for the treatment of Chronic myeloid leukemia₅₀280nM), the first generation Bcr/Abl kinase inhibitor, developed by noval pharmaceuticals and approved by the FDA in 2001 for the treatment of acute phase, accelerated phase and chronic phase of adult patients with philadelphia chromosome (Bcr/Abl) positive chronic myelogenous leukemia (CML for short) (Druker, b.j.; talpaz, m.; et.al.nat Med,1996,2,561-566.Deininger, m.w.; goldman, j.m.; al.blood,1997,90,3691-3698.Capdeville, R.; buchdinger, e.; et.al. nat. rev. drug discov.2002,1(7),493-502.Deininger, m.; buchdinger, e.; et. Al. blood 2005,105(7),2640-

Dasatinib (BMS 354825) is a second-generation Bcr/Abl kinase inhibitor, is a multi-target receptor tyrosine kinase inhibitor TKI developed by Behcet-MeishiGuibao company in the United states, is a substituted thiazole amide compound, is a dual inhibitor of powerful competitive Src and Bcr-Abl, and also has an inhibiting effect on C-kit, Btk, EPHA2 and PDGFR beta. Acting on Abl, Src and c-Kit, IC in cell-free assays₅₀0.6nM,0.8nM and 79nM, respectively (O' Hare, T.; Walters, D.K.; et. al. cancer Res 2005,65, 4500-. Dasatinib is much more effective than Imatinib in inhibiting proliferation of Ba/F3 of wild type Bcr-Abl or all Bcr-Abl mutants (except T315I). Dasatinib exhibits a 2-fold exponentially increasing potency (approximately 325-fold) compared to Imatinib. Dasatinib was approved by the FDA at 6 months 2006 for CML and Ph that were ineffective or intolerant to previous treatments (including imatinib)⁺Adult patients with Acute Lymphoblastic Leukemia (ALL). Studies have shown that (Walz, C.; Sattler, M.Crit Rev Oncol Hematol,2006,57(2),145-164.),the combination of Dasatinib and Abl has no strict configuration requirement, so that the composition has good curative effect on imatinib-resistant CML. Dasatinib can bind not only to Bcr-Abl proteins in the inactive state (DFG-out) but also to Bcr-Abl proteins in the active state (DFG-in). This binding pattern allows for greater binding adaptability and greater binding capacity of dasatinib, thus showing better inhibitory activity. Dasatinib is currently in clinical research for Chronic Lymphocytic Leukemia (CLL), breast and prostate cancer, and the like. In addition, Dasatinib also has inhibitory effects on Discoidin Domain Receptors (DDRs) because of the high similarity of DDRs to Bcr-Abl kinases in the sequence of the ATP-binding domain.

Discoidin Domain Receptors (DDRs) are a unique class of receptor tyrosine kinases, named for their unique extracellular N-terminal Discoidin receptor binding domains, and are the only transmembrane receptor tyrosine kinases known to date that use collagen as a signaling molecule. DDRs are involved in the regulation of a variety of intracellular physiological processes and have two members, DDR1 and DDR 2. There is increasing research evidence that DDR1 and DDR2 play important roles in normal physiological activities of cells, such as migration, proliferation, differentiation, etc. DDRs can be involved in a variety of normal physiological processes in cells, and thus when expression levels of such proteins become abnormal, they can also lead to dysplasia and cause a variety of related diseases. The DDRs can be used as potential targets for disease treatment, and more evidences indicate that the abnormal expression of the DDRs can be used as tumor markers for diagnosis and treatment of various tumors, such as lung cancer, breast cancer, liver cancer, pancreatic cancer, prostate cancer, cervical cancer and the like. In the non-small cell lung cancer sample, DDR1 overexpression level was significantly associated with tumor lymph node metastasis (p ═ 0.001). The molecular biological mechanism research shows that the increase of the expression level of DDR1 can promote the migration and invasion of lung cancer cells by promoting the EMT process. In breast cancer cells, the expression level of DDR2 was increased 6-fold compared to the normal level and showed close correlation with tumor development; while the expression level of DDR1 appears to be down-regulated. Multivariate analysis indicates that DDR2 can be used as an independent diagnostic marker for breast cancer treatment. The new evidence also indicates that DDR2 can promote tumor cells to transfer from mammary glands to lungs, and further shows that DDR2 has good application prospect in breast cancer diagnosis and treatment.

The discoid domain receptor is also a potential drug target, and the development of a discoid domain receptor small molecule inhibitor can play a role in the treatment field of diseases such as inflammation, hepatic fibrosis, renal fibrosis, pulmonary fibrosis, skin scar, atherosclerosis and the like. In fact, no discoidin domain receptor inhibitors are currently clinically used for the treatment of the above-mentioned diseases.

Immune cells can be generally divided into two categories, T cells and B cells, wherein the main role of B cells is to secrete various antibodies to help the body resist various foreign invasion. Bruton's tyrosine kinase (Btk) is a key kinase in the signal transduction pathway of B cell antigen receptor (Bcr), is mainly expressed in B cells, is distributed in lymphatic system, hemopoietic system and blood system, can regulate the maturation and differentiation of normal B cells, is also closely related to various B cell lymphoid tissue disorder diseases, and is a hot target for treating hematological malignancies and autoimmune disorder diseases at present. In recent years, researches on B cells, particularly on B cell non-Hodgkin lymphoma and rheumatoid arthritis show that Btk is often abnormally expressed. Ibrutinib (Ibrutinib) is currently the most attractive Btk targeted inhibitor, has significant therapeutic effects on various B cell tumors and autoimmune diseases in preclinical and clinical studies, shows outstanding therapeutic activity on mantle cell lymphoma and chronic lymphocytic leukemia, and is approved for marketing.

The receptor tyrosine kinase c-Kit (also named CD117) is a transmembrane receptor protein with tyrosine kinase activity coded by a retrovirus proto-oncogene c-Kit, forms a III type receptor tyrosine kinase superfamily together with a Platelet Derived Growth Factor Receptor (PDGFR), a macrophage colony stimulating factor-1 receptor (CSF-1R) and an Fms-like tyrosine kinase receptor 3(FLT3), plays an important role in the processes of occurrence, development, invasion, migration and relapse of tumors, is one of hot targets of the current tumor molecule targeted therapy, and an inhibitor of the receptor tyrosine kinase c-Kit also becomes an anti-tumor molecule targeted therapy targetThe hot spot of drug research and development. Dasatinib inhibits c-Kit kinase, IC in cell-free assays₅₀This was 79 nM.

Therefore, the micromolecule derived from the multi-target receptor tyrosine kinase inhibitor Dasatinib can be used as a tyrosine kinase inhibitor, can have inhibitory activity on kinases such as DDR1, DDR2, Abl, Src, Btk and Kit and has antiproliferative activity on K562 cells.

Currently, available Abl and Src dual tyrosine kinase inhibitor drugs include Dasatinib, Bosutinib and Ponatiniib. There are also molecules under investigation. DCC-2036 (Rebatib) is a Bcr-Abl inhibitor and acts on Abl1(WT) and Abl1(T315I), IC₅₀0.8nM and 4nM, respectively, also inhibit Src kinase, now in clinical phase I. Among the preclinical compounds studied were: NVP-BHG712, a specific EphB4 inhibitor, ED₅₀At 25nM, it also has inhibitory activity on c-Src and c-Abl, IC₅₀1.266 μ M and 1.667 μ M, respectively; PP1, also a potent Src selectivity inhibitor, acts on Lck/Fyn with IC₅₀5nM/6nM, IC for Bcr/Abl₅₀Is 1 mu M; PD173955, a potent Bcr-Abl inhibitor, IC₅₀1-2nM, and also inhibit Src activity, IC₅₀＝22nM。

However, there is still no good small molecule inhibitor for the above tyrosine kinase, and there is still a continuous effort from the developers.

Disclosure of Invention

In view of the above, it is necessary to provide a tyrosine kinase inhibitor having a good tyrosine kinase inhibitory effect.

A tyrosine kinase inhibitor having the structural features of formula I or a pharmaceutically acceptable salt or stereoisomer thereof:

wherein:

R₁，R₂independently selected from: C1-C6 alkyl, halogen substituted C1-C6 alkyl, C1-C6 alkylOxy, halogen-substituted C1-C6 alkoxy, C2-C6 alkenyl, halogen-substituted C2-C6 alkenyl, C2-C6 alkynyl, halogen-substituted C2-C6 alkynyl, C3-C6 cycloalkyl, halogen-substituted C3-C6 cycloalkyl, C1-C6-substituted carbonyl, amido and ester groups, a three-to six-membered heterocyclic ring containing N, O, S heteroatom, halogen, or H;

the five-membered cyclic group comprising A, B is selected from one of the following groups:

wherein: r₄，R₅，R₆Independently selected from: h, C1-C6 alkyl, C6-C10 substituted aralkyl, halogen substituted methyl, OR₇，NR₇R₈，CN，COOR₇，CONR₇R₈，SO₂R₇，SO₂NR₇R₈，NO₂，NCONR₇R₈，NCOOR₇，OCONR₇R₈，CSNR₇R₈，NCSNR₇R₈；

R₇、R₈Independently selected from: H. C1-C6 alkyl, N, S, O heteroatom substituted C1-C6 alkyl; v is selected from one of the following groups:

wherein: r₉Selected from: h, C1-C6 alkyl, halogen substituted C1-C6 alkyl, C1-C6 alkoxy, C6-C10 substituted aralkyl;

w is selected from one of the following groups:

m is selected from: 1 to 6;

x is selected from: o, S, N, or none;

R₃selected from: C1-C6 alkyl, halogen substituted C1-C6 alkyl, C1-C6 alkoxy, C1-C6 nitrogenous alkyl, C1-C6 alkyl substituted aryl,C4-C11 substituted acrylamide, aryl or one of the following substituents:

wherein: n is selected from: 1 to 6;

R₁₀，R₁₁，R₁₂independently selected from: h, C1-C6 alkyl, halogen substituted C1-C6 alkyl, C1-C6 alkoxy, C6-C10 substituted aralkyl.

In some of these embodiments, the compound is selected from compounds having the following structural features of formula II:

R₁，R₂，R₃v, W, X are as defined above.

In some of these embodiments, V is selected from one of the following groups:

w is selected from one of the following groups:

x is selected from: o, or none;

R₉as claimed in claim 1.

In some of these embodiments, R₉Selected from: h;

in some of these embodiments, R₃Selected from: C1-C6 alkyl, halogen substituted C1-C6 alkyl, C1-C6 nitrogen-containing alkyl, C1-C6 alkyl substituted aryl, aryl or the following substituents:

wherein: n is selected from: 1 to 6;

R₁₀selected from: H.

in some of these embodiments, X is selected from: o;

R₃selected from: C1-C6 alkyl, halogen substituted C1-C6 alkyl, C1-C6 nitrogenous alkyl, or N-ethylmorpholine.

In some of these embodiments, R₁，R₂Independently selected from: C1-C6 alkyl, or halogen; among them, the C1-C6 alkyl group is preferably methyl.

In some of these embodiments, one of the following compounds is selected:

methyl 4- (6- ((5- ((2-chloro-6-methylphenyl) carbamoyl) -2-thiazolyl) amino) -2-methyl-4-pyrimidinyl) -1-piperazine carboxylate;

ethyl 4- (6- ((5- ((2-chloro-6-methylphenyl) carbamoyl) -2-thiazolyl) amino) -2-methyl-4-pyrimidinyl) -1-piperazine carboxylate;

isopropyl 4- (6- ((5- ((2-chloro-6-methylphenyl) carbamoyl) -2-thiazolyl) amino) -2-methyl-4-pyrimidinyl) -1-piperazine carboxylate;

allyl 4- (6- ((5- ((2-chloro-6-methylphenyl) carbamoyl) -2-thiazolyl) amino) -2-methyl-4-pyrimidinyl) -1-piperazine carboxylate;

butyl 4- (6- ((5- ((2-chloro-6-methylphenyl) carbamoyl) -2-thiazolyl) amino) -2-methyl-4-pyrimidinyl) -1-piperazine carboxylate;

isobutyl 4- (6- ((5- ((2-chloro-6-methylphenyl) carbamoyl) -2-thiazolyl) amino) -2-methyl-4-pyrimidinyl) -1-piperazine carboxylate;

phenyl 4- (6- ((5- ((2-chloro-6-methylphenyl) carbamoyl) -2-thiazolyl) amino) -2-methyl-4-pyrimidinyl) -1-piperazine carboxylate;

benzyl 4- (6- ((5- ((2-chloro-6-methylphenyl) carbamoyl) -2-thiazolyl) amino) -2-methyl-4-pyrimidinyl) -1-piperazine carboxylate;

2-chloroethyl 4- (6- ((5- ((2-chloro-6-methylphenyl) carbamoyl) -2-thiazolyl) amino) -2-methyl-4-pyrimidinyl) -1-piperazine carboxylate;

2-morpholinoethyl 4- (6- ((5- ((2-chloro-6-methylphenyl) carbamoyl) -2-thiazolyl) amino) -2-methyl-4-pyrimidinyl) -1-piperazine carboxylate;

n, N-dimethylethyl 4- (6- ((5- ((2-chloro-6-methylphenyl) carbamoyl) -2-thiazolyl) amino) -2-methyl-4-pyrimidinyl) -1-piperazine carboxylate;

2- ((6- (4-propenoyl-1-piperazinyl) -2-methyl-4-pyrimidinyl) amino) -N- (2-chloro-6-methylphenyl) -5-thiazolecarboxamide;

(E) -N- (2-chloro-6-methylphenyl) -2- ((6- (4- (2-butenoyl) -1-piperazinyl) -2-methyl-4-pyrimidinyl) amino) -5-thiazolecarboxamide;

(E) -N- (2-chloro-6-methylphenyl) -2- ((6- (4- (4-dimethylamino) -2-butenoyl) -1-piperazinyl) -2-methyl-4-pyrimidinyl) amino) -5-thiazolecarboxamide;

(E) -N- (2-chloro-6-methylphenyl) -2- ((6- (4- (2-acrylamidoacetyl) -1-piperazinyl) -2-methyl-4-pyrimidinyl) amino) -5-thiazolecarboxamide;

(E) -N- (2-chloro-6-methylphenyl) -2- ((6- (4- (3-acrylamidopropionyl) -1-piperazinyl) -2-methyl-4-pyrimidinyl) amino) -5-thiazolecarboxamide;

(E) -N- (2-chloro-6-methylphenyl) -2- ((6- (4- (2- (2-acrylamidoacetylamino) acetyl) -1-piperazinyl) -2-methyl-4-pyrimidinyl) amino) -5-thiazolecarboxamide;

(E) -N- (2-chloro-6-methylphenyl) -2- ((6- (4- (2- (3-acrylamidopropionylamino) acetyl) -1-piperazinyl) -2-methyl-4-pyrimidinyl) amino) -5-thiazolecarboxamide;

(E) -N- (2-chloro-6-methylphenyl) -2- ((6- (4- (3- (2-acrylamidoacetylamino) propionyl) -1-piperazinyl) -2-methyl-4-pyrimidinyl) amino) -5-thiazolecarboxamide;

(E) -N- (2-chloro-6-methylphenyl) -2- ((6- (4- (3- (2- (2-butenamide) acetylamino) propionyl) -1-piperazinyl) -2-methyl-4-pyrimidinyl) amino) -5-thiazolecarboxamide;

(E) -N- (2-chloro-6-methylphenyl) -2- ((6- (4- (3- (3-acrylamidopropionylamino) propionyl) -1-piperazinyl) -2-methyl-4-pyrimidinyl) amino) -5-thiazolecarboxamide;

(E) -N- (2-chloro-6-methylphenyl) -2- ((6- (4- (3- (3- (2-butenamide) propionylamino) propionyl) -1-piperazinyl) -2-methyl-4-pyrimidinyl) amino) -5-thiazolecarboxamide;

(E) -N- (2-chloro-6-methylphenyl) -2- ((6- (4- (3- (3- (4-dimethylamino) -2-butenamide) propionylamino) propionyl) -1-piperazinyl) -2-methyl-4-pyrimidinyl) amino) -5-thiazolecarboxamide;

methyl 3- (6- ((5- ((2-chloro-6-methylphenyl) carbamoyl) -2-thiazolyl) amino) -2-methyl-4-pyrimidinyl) -1-pyrrolidinecarboxylic acid salt;

methyl 3- (6- ((5- ((2-chloro-6-methylphenyl) carbamoyl) -2-thiazolyl) amino) -2-methyl-4-pyrimidinyl) -1-pyrrolidinecarboxylic acid salt;

isopropyl 3- (6- ((5- ((2-chloro-6-methylphenyl) carbamoyl) -2-thiazolyl) amino) -2-methyl-4-pyrimidinyl) -1-pyrrolidine carboxylate;

allyl 3- (6- ((5- ((2-chloro-6-methylphenyl) carbamoyl) -2-thiazolyl) amino) -2-methyl-4-pyrimidinyl) -1-pyrrolidinecarboxylic acid salt;

butyl 3- (6- ((5- ((2-chloro-6-methylphenyl) carbamoyl) -2-thiazolyl) amino) -2-methyl-4-pyrimidinyl) -1-pyrrolidinecarboxylic acid salt;

isobutyl 3- (6- ((5- ((2-chloro-6-methylphenyl) carbamoyl) -2-thiazolyl) amino) -2-methyl-4-pyrimidinyl) -1-pyrrolidine carboxylate;

phenyl 3- (6- ((5- ((2-chloro-6-methylphenyl) carbamoyl) -2-thiazolyl) amino) -2-methyl-4-pyrimidinyl) -1-pyrrolidine carboxylate;

benzyl 3- (6- ((5- ((2-chloro-6-methylphenyl) carbamoyl) -2-thiazolyl) amino) -2-methyl-4-pyrimidinyl) -1-pyrrolidine carboxylate;

2- ((2-methyl-6- ((1- (methylsulfonyl) -3-pyrrolidinyl) amino) 4-pyrimidinyl) amino) -5-thiazolecarboxamide;

2- ((2-methyl-6- ((1- (ethylsulfonyl) -3-pyrrolidinyl) amino) 4-pyrimidinyl) amino) -2- (2-chloro-6-methylphenyl) -5-thiazolecarboxamide;

(E) -N- (2-chloro-6-methylphenyl) -2- ((6- ((1-acryloyl-3-pyrrolidinyl) amino) -2-methyl-4-pyrimidinyl) amino) -5-thiazolecarboxamide;

(E) -N- (2-chloro-6-methylphenyl) -2- ((6- ((1- (2-butenoyl) -3-pyrrolidinyl) amino) -2-methyl-4-pyrimidinyl) amino) -5-thiazolecarboxamide;

(E) -N- (2-chloro-6-methylphenyl) -2- ((6- ((1-propionyl-3-pyrrolidinyl) amino) -2-methyl-4-pyrimidinyl) amino) -5-thiazolecarboxamide.

The invention also discloses a preparation method of the tyrosine kinase inhibitor or the pharmaceutically acceptable salt or the stereoisomer thereof, which adopts the following route to synthesize:

or synthesized by adopting the following route:

the invention also discloses application of the tyrosine kinase inhibitor or the pharmaceutically acceptable salt or the stereoisomer thereof in preparing medicaments for preventing and treating tumors.

Compared with the prior art, the invention has the following beneficial effects:

according to the morphological structure of a protein composite crystal combined by Dasatinib and Abl, Src, Kit and other kinases and the high sequence homology of DDRs and Abl kinases, a class of small molecular compounds with structural characteristics of a formula I is designed and synthesized on the basis of Dasatinib, and experiments prove that the compounds can effectively inhibit the activity of tyrosine kinases. The compound has an inhibitory effect on kinases such as DDR1, DDR2, Abl, Src, Btk, Kit and the like, the half inhibitory concentration of the compound is compared with or superior to that of a positive control dasatinib, or at least equivalent to that of the positive control, particularly the compound 8j has a half inhibitory concentration which is less than that of the positive control on five tyrosine kinases such as DDR1, DDR2, Src, Btk and Kit, and K562 cells, and has good enzyme inhibitory activity and cell activity.

Detailed Description

As used herein, "alkyl" refers to a saturated hydrocarbon group or an unsaturated chain alkyl group, "chain alkyl" refers to a straight or branched chain alkyl group, such as C1-C6 chain alkyl refers to a saturated or unsaturated, straight or branched chain alkyl group having 1 to 6 carbon atoms, wherein examples of saturated straight chain alkyl groups include, but are not limited to, ethyl, n-propyl, and the like, examples of saturated branched chain alkyl groups include, but are not limited to, isopropyl, t-butyl, and the like, examples of unsaturated straight chain alkyl groups include, but are not limited to, vinyl, propenyl, and the like, and examples of unsaturated branched chain alkyl groups include, but are not limited to; "cycloalkyl" refers to an alkyl group having a cyclic structure, such as C3-C6 cycloalkyl refers to a saturated or unsaturated alkyl group having a cyclic structure having 3 to 6 carbon atoms, wherein examples of saturated cyclic alkyl groups include, but are not limited to, cyclopropyl, cyclopentyl, ethyl-substituted cyclohexyl, and the like, and examples of unsaturated cyclic alkyl groups include, but are not limited to, cyclopentene and the like.

The term "halogen" means F, Cl, Br, I and the like, preferably F, Cl, Br, particularly preferably F and Cl.

The term "alkoxy" refers to a group in which an alkyl group is directly attached to an oxygen, such as methoxy, ethoxy, and the like.

The term "substituted" refers to the replacement of a hydrogen radical in a particular structure with a radical of a specified substituent. In the general formula, the short line drawn from the outside of the ring into the ring indicates that the substituent is not fixed at the substitution position on the ring and can be substituted at a position allowed by any chemical bond.

The invention includes the free forms of the compounds of formulae I-II, as well as pharmaceutically acceptable salts and stereoisomers thereof. Pharmaceutically acceptable salts of the invention can be synthesized from compounds of the invention containing a basic or acidic moiety by conventional chemical methods. In general, salts of basic compounds are prepared by ion exchange chromatography or by reaction of the free base with a stoichiometric amount or excess of an inorganic or organic acid in the form of the desired salt in an appropriate solvent or combination of solvents. Similarly, salts of acidic compounds are formed by reaction with suitable inorganic or organic bases.

Thus, pharmaceutically acceptable salts of the compounds of the present invention include the conventional non-toxic salts of the compounds of the present invention formed by the reaction of a basic compound of the present invention and an inorganic or organic acid. For example, conventional non-toxic salts include those prepared from inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, sulfamic acid, phosphoric acid, nitric acid, and the like, as well as those prepared from organic acids such as acetic acid, propionic acid, succinic acid, glycolic acid, stearic acid, lactic acid, malic acid, tartaric acid, citric acid, ascorbic acid, pamoic acid, maleic acid, hydroxymaleic acid, phenylacetic acid, glutamic acid, benzoic acid, salicylic acid, sulfanilic acid, 2-acetoxy-monobenzoic acid, fumaric acid, toluenesulfonic acid, methanesulfonic acid, ethane disulfonic acid, oxalic acid, isethionic acid, trifluoroacetic acid, and the like.

If a compound of the invention is acidic, an appropriate "pharmaceutically acceptable salt" refers to a salt prepared by a pharmaceutically acceptable non-toxic base including inorganic and organic bases. Salts derived from inorganic bases include aluminum, ammonium, calcium, copper, ferric, ferrous, lithium, magnesium, manganic, manganous, potassium, sodium, zinc, and the like. Particularly preferred are ammonium, calcium, magnesium, potassium and sodium salts. Salts derived from pharmaceutically acceptable organic non-toxic bases including salts of primary, secondary and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines and basic ion exchange resins such as arginine, betaine, caffeine, choline, N' -dibenzylethylenediamine, diethylamine, 2-dimethylaminoethanol, aminoethanol, ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucosamine, histidine, hydroxycobalamin, isopropylamine, lysine, methylglucamine, morpholine, piperazine, piperidine, piperdine, polyamine resins, procaine, purines, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine and the like.

The compounds of the present invention can be prepared using methods in the following synthetic routes, in addition to standard methods known in the literature or exemplified in experimental procedures. The compounds and methods of synthesis described in the present invention can be better understood in conjunction with the synthetic schemes described below. The synthetic schemes describe the methods that can be used to prepare the compounds of the present invention, and the methods are described as illustrative schemes for illustrative purposes only and do not limit the scope of the present invention.

Example 1

Preparation of compound 8a, methyl 4- (6- ((5- ((2-chloro-6-methylphenyl) carbamoyl) -2-thiazolyl) amino) -2-methyl-4-pyrimidinyl) -1-piperazine carboxylate.

(1) Compound I-7 was synthesized according to the above scheme.

Wherein the compounds I-3, I-4, I-5 and I-6 are known compounds, and the Synthesis method is performed according to the literature (F. Fern < et al, Synthesis 2001, No.2, 239-242; Bang-Chi Chen et al ARKIVOC 2010(vi), 32-38).

Characterization data for compound I-3 are:¹H NMR(400MHz，CDCl₃)δppm7.60(d,J＝12.10Hz,1H)，5.32(d,J＝12.10Hz,1H),3.88(q,J＝7.10Hz,2H),1.20(t,J＝7.10Hz,3H).

characterization data for compound I-4 are:¹H NMR(400Hz,DMSO-d₆)δppm9.28(s,1H),7.45(d,J＝12.4Hz,1H),7.27-7.37(d,J＝7.5Hz,1H),7.10-7.27(m,J＝7.5Hz,2H),5.58(d,J＝12.4Hz,1H),3.94(q,J＝7Hz,2H),2.15(s,3H),1.26(t,J＝7Hz,3H).

characterization data for compound I-5 are:¹H NMR(400Hz,DMSO-d₆)δppm 9.63(s,1H)，7.85(s,1H),7.61(s,2H),7.29-7.43(d,J＝7.5,1H),7.09-7.29(m,J＝7.5,2H),2.19(s,3H).

characterization data for compound I-6 are:¹H NMR(400MHz,DMSO-d₆)δppm 12.24(br s,1H)，10.02(s,1H),8.32(s,1H),7.41(d,J＝7.7Hz,1H),7.30(dd,J＝6.1,6.7Hz,1H),7.26(d,J＝7.7Hz,1H),6.95(s,1H),2.59(s,3H),2.24(s,3H).

the synthesis method of the compound I-7 is as follows: in a 250mL two-necked flask equipped with a reflux condenser, Compound I-6(10.00g,25.4mmol) was added at room temperature, dissolved in freshly distilled DMF (N, N-dimethylformamide), 1-Boc-piperazine (6.6g,35.6mmol) was added, and Et3N (triethylamine, 7.14mL,50.8mmol) was slowly added under a nitrogen stream. Stirring for 1h, and then heating to 80 ℃ for reaction. TLC monitored the reaction. After the reaction is finished, the reaction product is cooled to room temperature, the solvent is dried in a spinning mode, water is added, solid is filtered by suction and washed by ice-cold water, and then the yellow product is obtained by recrystallization of the solid by using anhydrous methanol. The yellow product was transferred to 10mL of 30% TFA (trifluoroacetic acid) in dichloromethane and the reaction stirred at room temperature, monitored by TLC, ended, evaporated to dryness, the residue washed 3 times with saturated sodium bicarbonate solution, filtered off with suction, and the filter cake dried to give I-78.6g in 86% yield.

The characterization data for this compound I-7 are:¹H NMR(400MHz,DMSO-d₆)δppm 11.42(s,1H),9.87(s,1H),8.21(s,1H),7.41-7.25(m,3H),6.02(s,1H),3.45-3.36(m,5H),3.02-2.75(m,4H),2.40(s,3H),2.24(s,3H).ESI-MS m/z:[M+H]⁺＝444.1

(2) preparation of compound 8a, methyl 4- (6- ((5- ((2-chloro-6-methylphenyl) carbamoyl) -2-thiazolyl) amino) -2-methyl-4-pyrimidinyl) -1-piperazine carboxylate using compound I-7 with methyl chloroformate was as follows:

at 0 deg.C, in a 25mL single-necked flask, compound I-7(0.8mmol,1eq), NaHCO was added₃(1.6mmol,2eq), THF/water (5mL,4:1, v/v) as a mixed solvent was added, methyl chloroformate (1.2mmol,1.5eq) was slowly added, and after the addition, the reaction was carried out at 0 ℃ for 1 hour, and the reaction was carried out at room temperature. TLC monitoring, reaction was complete, 1mL of water was added to dilute the reaction, and the solvent was removed by rotary evaporation. The solid was washed with a little water and passed through the column to give the product 8a as a yellow solid in 77% yield.

The characterization data for this compound 8a are:¹H NMR(400MHz,DMSO-d₆)δ11.52(s,1H),9.89(s,1H),8.23(s,1H),7.39(d,J＝6.4Hz,1H),7.27(d,J＝9.2Hz,2H),6.07(s,1H),3.63(s,3H),3.56(s,4H),3.47(s,4H),2.42(s,3H),2.24(s,3H)；¹³C NMR(125MHz,DMSO-d₆)δ165.01,162.53,162.13,159.90,156.96,155.09,140.77,138.80,133.51,132.43,128.98,128.11,126.96,125.76,82.84,52.39,43.18,42.81,25.42,18.25.ESI-MS m/z:502.1(M+H)⁺,500.2(M-H)^-.。

the structural formula of the compound 8a is:

example 2

Preparation of compound 8b, ethyl 4- (6- ((5- ((2-chloro-6-methylphenyl) carbamoyl) -2-thiazolyl) amino) -2-methyl-4-pyrimidinyl) -1-piperazine carboxylate.

The procedure is as in example 1, using compound I-7 to react with ethyl chloroformate. Yellow solid 8b was obtained in 76% yield.

Characterization data for this compound 8b are:¹H NMR(400MHz,DMSO-d₆)δ10.03(s,1H),8.31(s,1H),7.39(d,J＝6.8Hz,1H),7.26(d,J＝9.3Hz,2H),6.21(s,1H),4.07(dd,J＝13.6,6.6Hz,2H),3.49(s,8H),2.48(s,3H),2.24(s,3H),1.20(t,J＝6.9Hz,3H)；¹³C NMR(125MHz,DMSO-d₆)δ163.64,162.51,161.29,160.25,159.57,156.99,156.49,154.70,138.80,133.40,132.44,129.07,128.27,127.04,83.69,61.02,44.02,42.15,24.14,18.32,14.59.ESI-MS m/z:516.2(M+H)⁺,514.1(M-H)^-。

the structural formula of the compound 8b is:

example 3

Preparation of compound 8c, isopropyl 4- (6- ((5- ((2-chloro-6-methylphenyl) carbamoyl) -2-thiazolyl) amino) -2-methyl-4-pyrimidinyl) -1-piperazine carboxylate.

The procedure is as in example 1, using compound I-7 and isopropyl chloroformate. Yellow solid 8c was obtained in 93% yield.

Characterization data for this compound 8c were:¹H NMR(400MHz,DMSO-d₆)δ9.99(s,1H),8.28(s,1H),7.40(d,J＝6.6Hz,1H),7.27(d,J＝9.6Hz,2H),6.18(s,1H),4.94-4.66(m,1H),3.59(s,4H),3.48(s,4H),2.47(s,3H),2.24(s,3H),1.21(d,J＝5.7Hz,6H)；¹³C NMR(125MHz,DMSO-d₆)δ163.61,160.32,159.32,158.19,156.49,154.26,138.74,137.83,134.37,133.37,132.38,128.99,128.18,126.97,83.45,68.23,43.95,42.52,24.15,21.95,18.27.ESI-MSm/z:530.2(M+H)⁺,528.2(M-H)^-.

the structural formula of compound 8c is:

example 4

Preparation of compound 8d, allyl 4- (6- ((5- ((2-chloro-6-methylphenyl) carbamoyl) -2-thiazolyl) amino) -2-methyl-4-pyrimidinyl) -1-piperazine carboxylate.

The preparation was carried out as in example 1, using compound I-7 and allyl chloroformate. Yellow solid 8d was obtained in 92% yield.

Characterization data for this compound 8d were:¹H NMR(400MHz,DMSO-d₆)δ12.04(s,1H),10.01(s,1H),8.29(s,1H),7.40(d,J＝7.2Hz,1H),7.30-7.25(m,2H),6.20(s,1H),5.95(ddd,J＝22.4,10.4,5.2Hz,1H),5.31(d,J＝17.2Hz,1H),5.21(d,J＝10.4Hz,1H),4.57(d,J＝5.2Hz,2H),3.62(s,4H),3.52(s,4H),2.48(s,3H),2.24(s,3H)；¹³C NMR(125MHz,DMSO-d₆)δ163.61,162.92,160.24,159.84,156.72,154.69,139.18,133.80,133.71,132.84,129.43,128.62,127.41,117.60,84.11,65.83,44.60,43.04,24.18,18.71.ESI-MS m/z:528.2(M+H)⁺,526.2(M-H)^-.

the structural formula of compound 8d is:

example 5

Preparation of compound 8e, butyl 4- (6- ((5- ((2-chloro-6-methylphenyl) carbamoyl) -2-thiazolyl) amino) -2-methyl-4-pyrimidinyl) -1-piperazine carboxylate.

The procedure is as in example 1, using compound I-7 and butyl chloroformate. Yellow solid 8e was obtained in 48% yield.

Characterization data for this compound 8e were:¹H NMR(400MHz,DMSO-d₆)δ11.63(s,1H),9.90(s,1H),8.24(s,1H),7.39(s,1H),7.28(s,2H),6.09(s,1H),4.03(s,1H),3.76(s,2H),3.55-3.47(m,4H),3.35(s,4H),3.18(s,2H),2.42(s,3H),2.24(s,3H),1.56-1.25(m,2H),1.13(m,3H)；¹³C NMR(125MHz,DMSO-d₆)δ165.33,165.17,162.25,157.18,156.98,154.71,140.84,138.79,133.52,132.43,128.98,128.13,126.96,125.90,82.82,64.63,43.25,42.29,30.58,25.52,18.63,18.27,13.59.ESI-MS m/z:543.2(M+H)⁺,542.2(M-H)^-.

the structural formula of compound 8e is:

example 6

Preparation of compound 8f, isobutyl 4- (6- ((5- ((2-chloro-6-methylphenyl) carbamoyl) -2-thiazolyl) amino) -2-methyl-4-pyrimidinyl) -1-piperazine carboxylate.

The procedure is as in example 1, using compound I-7 and isobutyl chloroformate. Yellow solid 8f was obtained in 78% yield.

Characterization data for this compound 8f were:¹H NMR(400MHz,DMSO-d₆)δ11.60(s,1H),9.94(s,1H),8.26(s,1H),7.40(d,J＝7.2Hz,1H),7.27(d,J＝10.8Hz,2H),6.12(s,1H),3.82(d,J＝6.4Hz,2H),3.57(s,8H),2.44(s,3H),2.24(s,3H),1.92-1.85(m,1H),0.90(d,J＝6.4Hz,6H)；¹³C NMR(125MHz,DMSO-d₆)δ164.52,162.50,161.45,159.75,156.76,154.69,140.49,138.77,133.48,132.42,128.95,128.09,126.94,125.89,83.09,70.81,43.47,34.08,27.50,24.94,18.84.ESI-MSm/z:543.2(M+H)⁺,542.2(M-H)^-.

the structural formula of compound 8f is:

example 7

Preparation of compound 8g, phenyl 4- (6- ((5- ((2-chloro-6-methylphenyl) carbamoyl) -2-thiazolyl) amino) -2-methyl-4-pyrimidinyl) -1-piperazine carboxylate.

The preparation was carried out as in example 1, using compound I-7 and phenyl chloroformate. 8g of a white solid are obtained in 94% yield.

Characterization data for this compound 8g is:¹H NMR(400MHz,DMSO-d₆)δ11.59(s,1H),9.92(s,1H),8.24(s,1H),7.40(d,J＝6.0Hz,3H),7.28-7.22(m,3H),7.16(d,J＝7.2Hz,2H),6.75(d,J＝7.6Hz,1H),6.13(s,1H),3.68(s,8H),2.45(s,3H),2.24(s,3H)；¹³C NMR(125MHz,DMSO-d₆)δ161.52,159.77,157.31,156.83,152.96,151.12,138.78,133.47,132.42,129.29,129.24,128.99,128.14,126.97,125.25,121.83,118.73,115.21,83.11,51.41,43.37,25.08,18.27.ESI-MS m/z:563.2(M+H)⁺,562.1(M-H)^-.

the structural formula of 8g of this compound is:

example 8

Preparation of compound 8h, benzyl 4- (6- ((5- ((2-chloro-6-methylphenyl) carbamoyl) -2-thiazolyl) amino) -2-methyl-4-pyrimidinyl) -1-piperazine carboxylate.

The procedure is as in example 1, using compound I-7 and benzyl chloroformate. A white solid was obtained in 8h, 77% yield.

Characterization data for this compound 8h are:¹H NMR(400MHz,DMSO-d₆)δ11.51(s,1H),9.88(s,1H),8.22(s,1H),7.40-7.20(m,8H),6.06(s,1H),6.13(s,1H),5.12(s,2H),3.56-3.52(m,8H),2.42(s,3H),2.34(s,3H)；¹³C NMR(125MHz,DMSO-d₆)δ161.21,162.54,162.27,159.92,157.01,154.50,140.82,138.81,136.81,133.53,132.45,128.985,128.78,128.41,128.13,127.85,127.59,126.98,125.77,82.84,66.35,43.16,25.52,18.27.ESI-MS m/z:578.2(M+H)⁺,576.2(M-H)^-.

the structural formula of the compound 8h is:

example 9

Preparation of the compound 8i, 2-chloroethyl 4- (6- ((5- ((2-chloro-6-methylphenyl) carbamoyl) -2-thiazolyl) amino) -2-methyl-4-pyrimidinyl) -1-piperazine carboxylate.

The procedure is as in example 1, using compound I-7 and 2-chloroethyl chloroformate. Yellow solid 8i was obtained in 88% yield.

Characterization data for this compound 8i are:¹H NMR(400MHz,DMSO-d₆)δ9.97(s,1H),8.26(s,1H),7.39(d,J＝6.8Hz,1H),7.30-7.23(m,2H),6.17(s,1H),4.29(t,J＝4.8Hz,2H),3.83(t,J＝4.8Hz,2H),3.61(s,8H),2.46(s,3H),2.24(s,3H)；¹³C NMR(125MHz,DMSO-d₆)δ163.98,162.59,162.46,162.37,160.78,159.62,154.21,138.80,133.40,132.42,129.06,128.25,127.03,83.40,65.03,43.73,43.00,42.76,24.53,18.30.ESI-MS m/z:550.1(M+H)⁺,548.2(M-H)^-.

the structural formula of the compound 8i is:

example 10

Preparation of the compound 8j, 2-morpholinoethyl 4- (6- ((5- ((2-chloro-6-methylphenyl) carbamoyl) -2-thiazolyl) amino) -2-methyl-4-pyrimidinyl) -1-piperazine carboxylate.

Compound 8i was reacted with morpholine. The specific operation is as follows: in a 50mL sealed tube, compound 8i (0.2mmol,1eq) was added, 5mL of DMF solvent was added, morpholine (0.4mmol,2eq) was added, the reaction was heated at 80 ℃ overnight, and TLC was used for detection. After the reaction, the solvent was dried by spinning, and the solid residue was passed through a column to give a yellow solid with a yield of 70%.

Characterization data for this compound 8j were:¹H NMR(400MHz,DMSO-d₆)δ9.87(s,1H),8.22(s,1H),7.39(d,J＝6.0Hz,1H),7.30-7.25(m,2H),6.06(s,1H),5.74(s,1H),4.14(t,J＝4.8Hz,2H),3.56(s,8H),3.47(s,8H),2.55(t,J＝4.8Hz,2H),2.42(s,3H),2.24(s,3H)；¹³C NMR(125MHz,DMSO-d₆)δ165.24,162.56,162.31,159.97,157.01,154.67,140.82,138.83,133.52,132.45,129.03,128.18,127.01,125.77,82.85,66.23,62.39,56.87,54.41,48.61,25.54,18.29.ESI-MS m/z:601.2(M+H)⁺,599.2(M-H)^-.

the structural formula of the compound 8j is:

example 11

Preparation of the compound 8k, N-dimethylethyl 4- (6- ((5- ((2-chloro-6-methylphenyl) carbamoyl) -2-thiazolyl) amino) -2-methyl-4-pyrimidinyl) -1-piperazine carboxylate.

Compound 8i was reacted with N, N-dimethylamine hydrochloride. The specific operation is as follows: in a 50mL sealed tube, compound 8i (0.2mmol,1eq) was added, DMF solvent 5mL was added, N-dimethylamine hydrochloride (0.4mmol,2eq) was added, triethylamine (0.4mmol,2eq) was added, the mixture was heated at 80 ℃ overnight for reaction, and TLC was used for detection. After the reaction, the solvent was dried by spinning, and the solid residue was passed through a column to give a yellow solid with a yield of 70%.

Characterization data for this compound 8k is:¹H NMR(400MHz,DMSO-d₆)δ11.52(s,1H),9.90(s,1H),8.24(s,1H),7.39(d,J＝7.2Hz,1H),7.26(q,J＝7.2Hz,2H),6.08(s,1H),4.13(t,J＝5.4Hz,2H),3.55(s,4H),3.48(s,4H),2.55(t,J＝5.4Hz,2H),2.42(s,3H),2.24(s,3H),2.22(s,9H)；¹³C NMR(125MHz,DMSO-d₆)δ163.61,160.32,159.32,158.19,156.49,154.26,138.74,137.83,134.37,133.37,132.38,128.99,128.18,126.97,83.45,68.23,43.95,42.52,24.15,21.95,18.27.ESI-MSm/z:558.2(M+H)⁺,557.2(M-H)^-.

the structural formula of compound 8k is:

example 12

Preparation of compound 8l, N- (2-chloro-6-methylphenyl) -2- ((6- (4-acryloyl-1-piperazinyl) -2-methyl-4-pyrimidinyl) amino) -5-thiazolecarboxamide, was carried out as follows:

under nitrogen protection, compound I-7(443mg,1mmol), dried DMF6mL, acryloyl chloride (100. mu.l, 1.2mmol), and triethylamine (0.6mL,2mmol) were added to a 25mL clean dry reaction flask, the reaction was stirred at 0 deg.C, monitored by TLC, upon completion of the reaction, the solvent was spun off, washed with saturated aqueous sodium bicarbonate, filtered to give a solid, which was passed over the column to give 195mg of product I-7 in 39% yield.

Characterization data for this compound 8i were:¹H NMR(400MHz,DMSO-d₆)δppm 11.50(s,1H),9.87(s,1H),8.22(s,1H),7.45-7.21(m,3H),6.83(dd,J＝16.7,10.5Hz,1H),6.47-5.94(m,1H),5.95-5.60(m,1H),4.32(d,J＝4.1Hz,1H),3.74-3.54(m,8H),2.43(s,3H),2.24(s,3H)；¹³C NMR(125MHz,DMSO-d₆)δ165.15,164.36,162.48,162.17,159.85,156.94,140.77,138.76,133.48,132.39,128.94,128.09,127.47,126.93,125.72,82.75,61.95,54.82,25.49,25.42,18.22.MS(ESI)m/z:499.0(M+H)⁺,497.0(M-H)^-.

the structural formula of this compound 8l is:

example 13

Preparation of compound 8m, (E) -N- (2-chloro-6-methylphenyl) -2- ((6- (4- (2-butenoyl) -1-piperazinyl) -2-methyl-4-pyrimidinyl) amino) -5-thiazolecarboxamide.

The objective product was obtained by a similar synthetic procedure to that of compound 8l in reference example 12, and reaction of compound I-7 with 2-butenoyl chloride gave 8m as a brown solid in 88% yield.

Characterization data for this compound 8m is:¹H NMR(400MHz,DMSO-d₆)δppm 11.49(s,1H),9.88(s,1H),8.22(s,1H),7.39(d,J＝7.2Hz,1H),7.26(dd,J＝7.2,6.8Hz,2H),6.72(td,J＝13.6,6.4Hz,1H),6.51(d,J＝14.8Hz,1H),6.07(s,1H),3.63(s,br,4H),3.57(s,4H),2.42(s,3H),2.24(s,3H),1.85(d,J＝6.4Hz,1H)；¹³C NMR(125MHz,DMSO-d₆)δ165.63,164.94,162.95,162.66,160.35,157.39,141.36,141.23,139.23,133.94,132.86,129.42,128.56,127.40,126.18,122.26,83.21,54.87,48.56,25.55,18.27,17.77.MS(ESI)m/z:513.0(M+H)⁺,511.0(M-H)^-.

the structural formula of the compound 8m is:

example 14

The compound 8N, (E) -N- (2-chloro-6-methylphenyl) -2- ((6- (4- (4-dimethylamino) -2-butenoyl) -1-piperazinyl) -2-methyl-4-pyrimidinyl) amino) -5-thiazolecarboxamide. The preparation method comprises the following steps:

compound I-7(100mg,0.23mmol), N, N-dimethyl-2-butanoate hydrochloride (45mg,0.27mmol), EDCI (1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride, 67.1mg,0.35mmol), HOBT (1-hydroxybenzotriazole, 47.5mg,0.35mmol), TEA (triethylamine, 0.06mL,0.46mmol), catalytic amounts of DMAP (4-dimethylaminopyridine) were added to a 50mL single-neck flask at room temperature, 3mL of dried DMF solvent was added, stirred at 0 deg.C, and after 30min, the mixture was allowed to react at room temperature. TLC monitoring, reaction completion, solvent spin-drying, washing with saturated aqueous sodium bicarbonate solution, generation of solid, filtration to obtain solid, and column chromatography to obtain white solid product 8n 58mg, yield 45.6%.

Characterization data for this compound 8n are:¹H NMR(400MHz,DMSO-d₆)δppm 9.89(s,1H),8.23(s,1H),7.38(s,1H),7.29-7.23(m,2H),7.17-7.13(m,1H),6.69-6.59(m,2H),6.07(s,1H),3.63(s,br,8H),3.05(s,2H),2.42(s,3H),2.28(t,J＝7.2Hz,6H),2.24(s,3H).¹³C NMR(125MHz,DMSO-d₆)δ165.63,164.94,162.95,162.66,160.35,157.39,141.36,141.23,139.23,133.94,132.86,129.42,128.56,127.40,126.18,122.26,83.21,54.87,48.56,25.55,18.27,17.77.MS(ESI)m/z:556.1(M+H)⁺,554.0(M-H)^-.

the structural formula of the compound 8n is:

example 15

Preparation of compound 8o, (E) -N- (2-chloro-6-methylphenyl) -2- ((6- (4- (2-acrylamidoacetyl) -1-piperazinyl) -2-methyl-4-pyrimidinyl) amino) -5-thiazolecarboxamide.

(1) Synthesis of Compound I-7-1.

Under the protection of nitrogen, I-7(443mg,1mmol), Boc-glycine (215mg,1.2mmol), HOBT (1-hydroxybenzotriazole, 203mg,1.5mmol), dried DMF 10mL, and catalytic amount of DMAP (4-dimethylaminopyridine) were added to a 25mL clean dry reaction flask, stirred in an ice-water bath for 5min, EDCI (1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride, 288mg,1.5mmol) was added, stirred for 30min, allowed to naturally rise to room temperature, stirred for further reaction, monitored by TLC, the reaction was complete, the solvent was spun dry, washed with saturated aqueous sodium bicarbonate, filtered to obtain a solid, and passed through a column to obtain the product. The product was transferred to 10mL of 30% TFA in dichloromethane and the reaction stirred at room temperature, monitored by TLC, ended, evaporated to dryness, the residue washed 3 times with saturated sodium bicarbonate solution, filtered and the filter cake dried to give I-7-1 as 341mg, 68% yield.

The characterization data of the compound I-7-1 are:¹H NMR(400MHz,DMSO-d₆)δppm 9.87(s,1H),8.22(s,1H),7.39(d,J＝7.2Hz,1H),7.24(t,J＝7.2Hz,2H),6.06(s,1H),3.57(s,br,6H),3.49(s,2H),3.37(s,5H),2.42(s,3H),2.24(s,3H).

the structural formula of the compound I-7-1 is as follows:

(2) preparation of compound 8o, (E) -N- (2-chloro-6-methylphenyl) -2- ((6- (4- (2-acrylamidoacetyl) -1-piperazinyl) -2-methyl-4-pyrimidinyl) amino) -5-thiazolecarboxamide.

The objective product was obtained by reacting compound I-7-1 with acryloyl chloride according to a similar synthetic procedure to compound 8l in example 12. White solid, 50mg, yield 87.4%.

Characterization data for this compound 8o is:¹H NMR(400MHz,DMSO-d₆)δppm 11.53(s,1H),9.88(s,1H),8.23(s,2H),7.39(s,1H),7.27(s,2H),6.39(s,1H),6.08(s,2H),5.61(s,1H),4.09(s,2H),3.58(s,8H),2.43(s,3H),2.24(s,3H)；¹³C NMR(125MHz,DMSO-d₆)167.64,165.78,165.74,162.99,160.61,157.26,141.25,139.21,133.73,132.79,131.54,129.47,128.71,127.40,126.46,126.07,83.26,43.67,41.33,40.91,25.84,18.59.MS(ESI)C₂₅H₂₇ClN₈O₃S,requires:555.05,Found:556.0(M+H)⁺,554.0(M-H)^-.

the structural formula of compound 8o is:

example 16

Preparation of compound 8p, (E) -N- (2-chloro-6-methylphenyl) -2- ((6- (4- (3-acrylamidopropionyl) -1-piperazinyl) -2-methyl-4-pyrimidinyl) amino) -5-thiazolecarboxamide.

(1) Synthesis of Compound I-7-2.

The method is the same as the synthesis of I-7-1, and the synthesis method comprises the steps of condensing I-7 and Boc-3-aminopropionic acid and removing Boc. The yield was 38.8%.

The characterization data of the compound I-7-2 are:¹H NMR(400MHz,DMSO-d₆)δppm 9.88(s,1H),8.22(s,1H),7.40(d,J＝6.4Hz,1H),7.27(d,J＝8.4Hz,2H),6.07(s,1H),3.84(s,8H),3.17(s,2H),2.77(s,1H),2.42(s,3H),2.24(s,3H).

the structural formula of the compound I-7-2 is as follows:

(2) preparation of compound 8p, (E) -N- (2-chloro-6-methylphenyl) -2- ((6- (4- (3-acrylamidopropionyl) -1-piperazinyl) -2-methyl-4-pyrimidinyl) amino) -5-thiazolecarboxamide.

The objective product was obtained by reacting compound I-7-2 with acryloyl chloride according to a similar synthetic procedure to compound 8l in example 12. White solid, 42mg, yield 81.5%.

Characterization data for this compound 8p were:¹H NMR(400MHz,DMSO-d₆)δppm 11.50(s,1H),9.87(s,1H),8.22-8.15(m,2H),7.39(d,J＝6.8Hz,1H),7.27(d,J＝10.0Hz,2H),6.23(dd,J＝17.2,10.0Hz,1H),6.07(d,J＝8.4Hz,2H),5.56(d,J＝9.6Hz,1H),3.56(s,8H),2.42(s,3H),2.24(s,3H)；¹³C NMR(125MHz,DMSO-d₆)δ169.51,165.36,164.94,162.67,162.37,160.11,157.07,140.94,138.93,133.59,132.54,131.79,129.14,128.31,127.11,125.86,125.24,82.92,44.25,43.35,35.21,32.51,25.64,18.37.MS(ESI)C₂₆H₂₉ClN₈O₃S requires:569.08,Found:570.1(M+H)⁺,568.0(M-H)^-.

the structural formula of the compound 8p is:

example 17

Preparation of compound 8q, (E) -N- (2-chloro-6-methylphenyl) -2- ((6- (4- (2- (2-acrylamidoacetylamino) acetyl) -1-piperazinyl) -2-methyl-4-pyrimidinyl) amino) -5-thiazolecarboxamide.

Compound I-7-3 was synthesized first, the route is as follows:

in a 50mL single neck flask, glycine ethyl ester hydrochloride (1.38g,10mmol), EDCI (1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride, 3.84g, 20mmol), a catalytic amount of DMAP (4-dimethylaminopyridine), about 20mL of DCM, TEA (triethylamine, 4.2mL,30mmol) and Boc-glycine (1.75g,10mmol) were added with stirring. The reaction was stirred and monitored by TLC. After the reaction is finished, adding water for washing, separating an organic phase, and then using saturated NH₄The solution is washed with Cl, the organic phase is dried by spinning, 20mL of ethanol is added, the mixture is stirred, aqueous NaOH solution (1g of NaOH,15mL of water) is added, and the hydrolysis is carried out under stirring at room temperature. TLC monitoring, after hydrolysis, add aqueous HCl (2N) at 0 ℃ to adjust the pH to about 4. And (3) a spin-drying system contains more salt and is directly used for the next reaction to synthesize a compound I-7-4: the method is the same as the synthesis of I-7-2, condensation is carried out on I-7 and I-7-3, and then the Boc is removed to obtain a product I-7-4. White solid, two-step yield 65.3%.

The structural formula of the compound I-7-4 is as follows:

finally 8q is synthesized. The objective product was obtained by reacting compound I-7-4 with acryloyl chloride according to a similar synthetic procedure to compound 8l in example 12. White product, 45mg, yield 91.2%.

Characterization data for this compound 8q is:¹H NMR(400MHz,DMSO-d₆)δppm 11.52(s,1H),9.88(s,1H),8.24(d,J＝15.2Hz,2H),7.69(d,J＝14.4Hz,1H),7.40(d,J＝6.4Hz,1H),7.27(d,J＝10.0Hz,2H),6.39(dd,J＝16.8,10.4Hz,1H),6.10(d,J＝22.0Hz,2H),5.61(d,J＝9.6Hz,1H),4.44-3.91(m,4H),3.58(s,8H),2.43(s,3H),2.24(s,3H)；¹³C NMR(125MHz,DMSO-d₆)δ167.30,167.15,165.40,165.07,162.38,160.17,157.08,138.94,133.59,132.55,131.81,131.64,131.53,129.15,128.76,128.33,127.12,125.74,82.96,65.18,43.43,41.00,40.63,25.63,18.37.MS(ESI)C₂₇H₃₀ClN₉O₄S requires:612.10,Found:613.1(M+H)⁺,611.1(M-H)^-.

the structural formula of the compound 8q is:

example 18

Preparation of compound 8r, (E) -N- (2-chloro-6-methylphenyl) -2- ((6- (4- (2- (3-acrylamidopropionamido) acetyl) -1-piperazinyl) -2-methyl-4-pyrimidinyl) amino) -5-thiazolecarboxamide.

Compound I-7-5 was synthesized first, the route is as follows:

the method is the same as the synthesis of I-7-3, condensation is carried out by Boc-3-aminopropionic acid and glycine ethyl ester hydrochloride, and hydrolysis is carried out by NaOH. The crude product I-7-5 is obtained and is directly used for the next step of synthesizing I-7-6. The method is the same as the synthesis of I-7-4, condensation is carried out on I-7 and I-7-5, and then the product I-7-6 is obtained by removing Boc. White solid, two-step yield 45.6%.

The characterization data for this compound I-7-6 are:¹H NMR(400MHz,DMSO-d₆)δppm 9.88(s,1H),8.22(s,1H),8.09(d,J＝34.2Hz,1H),7.40(d,J＝6.4Hz,1H),7.27(d,J＝9.6Hz,2H),6.07(s,1H),4.00(s,2H),3.56(s,12H),2.42(s,3H),2.24(s,3H).

the structural formula of the compound I-7-6 is as follows:

finally, 8r was synthesized by reacting compound I-7-6 with acryloyl chloride in a similar manner to the synthesis of compound 8l in example 12 to give the desired product. White product, 53mg, yield 98.1%.

Characterization data for this compound 8r are:¹H NMR(400MHz,DMSO-d₆)δppm 11.53(s,1H),9.88(s,1H),8.22(s,1H),8.13-8.09(m,2H),7.40(d,J＝6.8Hz,1H),7.33-7.21(m,2H),6.21(dd,J＝17.0,10.0Hz,1H),6.13-6.00(m,2H),5.56(d,J＝9.6Hz,1H),4.00(s,2H),3.56(s,8H),2.43(s,3H),2.41-2.28(m,4H),2.24(s,3H)；¹³C NMR(125MHz,DMSO-d₆)δ171.28,167.74,165.59,165.26,162.85,162.52,160.38,157.17,141.10,139.01,133.66,132.67,131.78,129.31,128.92,128.51,127.25,125.97,125.65,83.11,71.49,40.78,35.68,35.33,25.72,18.47.MS(ESI)C₂₈H₃₂ClN₉O₄Srequires:626.13,Found:627.1(M+H)⁺,625.0(M-H)^-.

the structural formula of compound 8r is:

example 19

Preparation of compound 8s, (E) -N- (2-chloro-6-methylphenyl) -2- ((6- (4- (3- (2-acrylamidoacetylamino) propionyl) -1-piperazinyl) -2-methyl-4-pyrimidinyl) amino) -5-thiazolecarboxamide.

Compound I-7-7 was synthesized first, the route is as follows:

the method is the same as the synthesis of I-7-3, condensation is carried out by Boc-glycine and 3-amino ethyl propionate hydrochloride, and hydrolysis is carried out by NaOH. The crude product I-7-7 is obtained and is directly used for the next step of synthesizing I-7-8. The method is the same as the synthesis of I-7-4, condensation is carried out on I-7 and I-7-7, and then the Boc is removed to obtain a product I-7-8. White solid, two-step yield 37.4%.

The characterization data for this compound I-7-8 are:¹H NMR(400MHz,DMSO-d₆)δppm 9.88(s,1H),8.22(s,1H),7.95(s,1H),7.40(d,J＝6.8Hz,1H),7.32-7.19(m,2H),6.06(s,1H),3.55(s,10H),3.05(s,2H),2.54-2.53(m,2H),2.42(s,3H),2.24(s,3H).

the structural formula of the compound I-7-8 is as follows:

finally 8s were synthesized by reacting compound I-7-8 with acryloyl chloride, using a procedure similar to that for the synthesis of compound 8l in example 12, to give the desired product. White product, 47mg, yield 86.3%.

Characterization data for this compound 8s are:¹H NMR(400MHz,DMSO-d₆)δppm 11.51(s,1H),9.88(s,1H),8.34(s,1H),8.22(s,1H),7.93(s,1H),7.39(s,1H),7.28(s,2H),6.37-6.24(m,1H),6.08(d,J＝17.2Hz,2H),5.60(d,J＝8.8Hz,1H),3.75(s,4H),3.55(s,10H),2.42(s,3H),2.23(s,3H)；¹³C NMR(125MHz,DMSO-d₆)δ169.63,168.97,165.41,165.24,162.70,162.40,160.17,157.07,140.97,138.96,133.59,132.56,131.52,129.17,128.35,127.13,125.78,82.94,44.31,43.57,43.29,42.23,40.46,35.22,32.55,25.64,18.38.MS(ESI)C₂₈H₃₂ClN₉O₄S requires:626.13,Found:627.1(M+H)⁺,625.1(M-H)^-.

the structural formula of the compound 8s is:

example 20

Preparation of compound 8t, (E) -N- (2-chloro-6-methylphenyl) -2- ((6- (4- (3- (2- (2-butenamide) acetylamino) propionyl) -1-piperazinyl) -2-methyl-4-pyrimidinyl) amino) -5-thiazolecarboxamide.

The objective product was obtained by reacting compound I-7-8 with 2-butenoyl chloride according to a similar synthetic procedure to compound 8l in example 12. White product, 50mg, 74.3% yield.

Characterization data for this compound 8t were:¹H NMR(400MHz,DMSO-d₆)δppm 11.58(s,1H),10.05(s,1H),8.32(s,1H),8.23(s,1H),7.95(s,1H),7.36(d,J＝7.2Hz,1H),7.31-7.17(m,2H),6.60(dt,J＝13.8,6.9Hz,1H),6.13(s,1H),5.99(d,J＝15.3Hz,1H),3.70(d,J＝5.5Hz,2H),3.55(s,8H),3.30-3.29(m,4H),2.42(s,3H),2.24(s,3H),1.78(d,J＝6.4Hz,3H)；¹³C NMR(125MHz,DMSO-d₆)δ169.52,169.11,165.38,165.25,162.53,162.29,160.08,157.06,141.07,138.89,138.28,133.64,132.55,129.04,128.19,127.02,125.82,125.67,82.96,44.25,43.54,43.19,35.13,32.49,25.53,18.37,17.38.MS(ESI)C₂₉H₃₄ClN₉O₄S requires:640.16,Found:641.2(M+H)⁺,639.2(M-H)^-.

the structural formula of the compound 8t is:

example 21

Preparation of compound 8u, (E) -N- (2-chloro-6-methylphenyl) -2- ((6- (4- (3- (3-acrylamidopropionamido) propionyl) -1-piperazinyl) -2-methyl-4-pyrimidinyl) amino) -5-thiazolecarboxamide.

Compound I-7-9 was synthesized first, the route is as follows:

the method is the same as the synthesis of I-7-3, Boc-3-aminopropionic acid and 3-aminopropionic acid ethyl ester hydrochloride are condensed, and then NaOH is used for hydrolysis to obtain a crude product I-7-9 which is directly used for synthesizing I-7-10 in the next step. The method is the same as the synthesis of I-7-4, condensation is carried out on I-7 and I-7-9, and then the Boc is removed to obtain a product I-7-10. White solid, two-step yield 65.9%.

The characterization data for this compound I-7-10 are:¹H NMR(400MHz,DMSO-d₆)δppm 9.88(s,1H),8.22(s,1H),7.95(s,1H),7.39(d,J＝6.8Hz,1H),7.34-7.19(m,2H),6.07(s,1H),3.55(s,12H),3.28(d,J＝4.4Hz,4H),2.42(s,3H),2.23(s,3H).

the structural formula of the compound I-7-10 is as follows:

finally 8u was synthesized by reacting compound I-7-10 with acryloyl chloride in a similar manner to the synthesis of compound 8l in example 12 to give the desired product. White product, 48mg, 88.2% yield.

Characterization data for this compound 8u are:¹H NMR(400MHz,DMSO-d₆)δppm 11.51(s,1H),9.88(s,1H),8.22(s,1H),8.11(s,1H),7.93(s,1H),7.40(d,J＝6.4Hz,1H),7.27(d,J＝8.8Hz,2H),6.21(dd,J＝16.8,10.0Hz,1H),6.05(d,J＝12.4Hz,2H),5.55(d,J＝9.6Hz,1H),3.55(s,10H),3.30-3.17(m,4H),2.42(s,3H),2.25(s,2H),2.24(s,3H)；¹³C NMR(125MHz,DMSO-d₆)δ170.43,169.46,165.26,164.70,162.57,162.28,159.99,156.99,140.84,138.84,133.52,132.46,131.76,129.04,128.19,127.01,125.77,124.99,82.83,44.18,43.49,43.18,35.32,35.03,32.61,25.55,18.20.MS(ESI)C₂₉H₃₄ClN₉O₄S requires:646.16,Found:647.2(M+H)⁺,645.1(M-H)^-.

the structural formula of the compound 8u is:

example 22

Preparation of compound 8v, (E) -N- (2-chloro-6-methylphenyl) -2- ((6- (4- (3- (3- (2-butenamide) propionylamino) propionyl) -1-piperazinyl) -2-methyl-4-pyrimidinyl) amino) -5-thiazolecarboxamide.

The objective product was obtained by reacting compound I-7-10 with 2-butenoyl chloride according to a similar synthetic procedure to compound 8l in example 12. Yellow solid, 45mg, yield 66.7%.

Characterization data for this compound 8v were:¹H NMR(400MHz,DMSO-d₆)δppm 11.52(s,1H),9.91(s,1H),8.24(s,1H),8.10-7.83(m,2H),7.40(d,J＝7.2Hz,1H),7.31-7.22(m,2H),6.58(dq,J＝13.8,6.8Hz,1H),6.08(s,1H),5.89(d,J＝14.2Hz,1H),3.55(s,8H),3.28(dd,J＝12.9,6.7Hz,5H),2.42(s,4H),2.23(d,J＝8.1Hz,6H),1.76(d,J＝6.6Hz,3H)；¹³C NMR(125MHz,DMSO-d₆)δ169.68,168.76,165.26,163.90,162.47,162.28,158.79,156.70,140.54,138.82,133.62,132.47,131.66,129.00,128.29,127.04,125.67,124.89,82.44,44.28,43.39,43.28,36.52,36.08,32.61,25.55,18.20,17.86.MS(ESI)C₃₀H₃₆ClN₉O₄S requires:654.18,Found:655.1(M+H)⁺,653.1(M-H)^-.

the structural formula of compound 8v is:

example 23

Preparation of compound 8w, (E) -N- (2-chloro-6-methylphenyl) -2- ((6- (4- (3- (3- (4-dimethylamino) -2-butenamide) propionylamino) propionyl) -1-piperazinyl) -2-methyl-4-pyrimidinyl) amino) -5-thiazolecarboxamide.

The objective product was obtained by reacting the compound 1-7-10 with N, N-dimethyl-2-butanoic acid hydrochloride according to a similar synthetic procedure to the compound 8N in reference example 14. Yellow solid, 53mg, yield 55.1%.

Characterization data for this compound 8w are:¹H NMR(400MHz,DMSO-d₆)δppm 11.50(s,1H),9.90(s,1H),8.24(s,1H),8.08-8.05(m,1H),7.95-7.92(m,1H),7.39(d,J＝7.6Hz,1H),7.30-7.23(m,2H),6.54(dq,J＝12.8,6.4Hz,1H),6.08-6.03(m,1H),5.31(s,1H),3.55(s,10H),2.42(s,3H),2.27(s,8H),2.24(s,3H)；¹³C NMR(125MHz,DMSO-d₆)δ169.70,168.96,165.46,163.96,162.67,162.30,158.86,156.54,140.34,138.76,133.62,132.50,131.76,129.10,128.30,127.00,125.58,124.77,82.44,64.20,46.58,46.19,45.28,37.68,36.52,36.08,32.44,25.59,18.17.MS(ESI)C₃₂H₄₁ClN₁₀O₄S requires:697.25,Found:698.2(M+H)⁺,696.2(M-H)^-.

the structural formula of compound 8w is:

example 24

Preparation of compound 10a, methyl 3- (6- ((5- ((2-chloro-6-methylphenyl) carbamoyl) -2-thiazolyl) amino) -2-methyl-4-pyrimidinyl) -1-pyrrolidinecarboxylic acid salt.

(1) Synthesis of Compound I-8.

In a 250mL two-necked flask equipped with a reflux condenser, Compound I-6(10.00g,25.4mmol) was added at room temperature, dissolved in DMSO, 1-Boc-3-aminopyrrolidine (6.6g,35.6mmol) was added, and DIPEA (N, N-diisopropylethylamine), 8.4mL,50.8mmol, was slowly added under a nitrogen stream. Stirring for 1h, and then heating to 80 ℃ for reaction. TLC monitored the reaction. After the reaction was complete, it was cooled to room temperature, water was added, the solid was filtered with suction and washed with ice cold water, and slurried with DCM to give the white pure intermediate. The intermediate was transferred to 10mL of 30% TFA in dichloromethane and the reaction stirred at room temperature, monitored by TLC, ended, evaporated to dryness, the residue washed 3 times with saturated sodium bicarbonate solution, filtered and the filter cake dried to give 7.7g of I-8 in 68% yield.

The characterization data for this compound I-8 are:¹H NMR(400MHz,DMSO-d₆)δ9.85(s,1H),8.20(s,1H),7.39(d,J＝7.0Hz,1H),7.26(d,J＝9.9Hz,2H),5.87(s,1H),2.99(d,J＝10.3Hz,1H),2.88(d,J＝6.9Hz,1H),2.78(s,1H),2.60(d,J＝6.9Hz,1H),2.36(s,3H),2.24(s,3H),2.03-1.91(m,1H),1.58(s,1H).

the structural formula of the compound I-8 is as follows:

(2) preparation of compound 10a, methyl 3- (6- ((5- ((2-chloro-6-methylphenyl) carbamoyl) -2-thiazolyl) amino) -2-methyl-4-pyrimidinyl) -1-pyrrolidinecarboxylic acid salt.

The procedure of example 1 was repeated except for using compound I-8 and methyl chloroformate. Yellow solid 10a was obtained in 45% yield.

Characterization of this Compound 10aThe data are as follows:¹H NMR(400MHz,DMSO-d₆)δ11.45(s,1H),9.91(s,1H),8.24(s,1H),7.40(s,1H),7.27(s,2H),5.94(s,1H),4.37(s,1H),3.58(s,3H),3.42(s,2H),3.19(s,1H),3.14(s,1H),2.38(s,3H),2.24(s,3H),2.10(s,1H),1.85(s,1H)；¹³C NMR(125MHz,DMSO-d₆)δ167.73,165.23,162.64,162.24,159.83,154.58,141.14,138.76,133.61,132.57,128.76,128.02,127.05,125.52,52.01,51.45,45.30,44.24,43.72,30.96,29.83,18.22.ESI-MS m/z:502.1(M+H)⁺,500.1(M-H)^-.

the structural formula of the compound 10a is:

example 25

Preparation of compound 10b, methyl 3- (6- ((5- ((2-chloro-6-methylphenyl) carbamoyl) -2-thiazolyl) amino) -2-methyl-4-pyrimidinyl) -1-pyrrolidinecarboxylic acid salt.

The procedure is as in example 1, using compound I-8 to react with ethyl chloroformate. A brown solid 10b was obtained in 36% yield.

Characterization data for this compound 10b were:¹H NMR(400MHz,DMSO-d₆)δ11.45(s,1H),9.91(s,1H),8.24(s,1H),7.40(s,1H),7.27(s,2H),5.90(d,J＝30.6Hz,1H),4.02(s,2H),3.57(s,2H),3.42(s,2H),3.18(s,1H),3.16(s,1H),2.38(s,3H),2.24(s,3H),2.10(s,1H),1.84(s,1H),1.19(s,3H)；¹³C NMR(125MHz,DMSO-d₆)δ165.37,162.68,162.50,159.94,155.89,154.19,140.90,138.79,133.50,132.44,128.93,128.04,126.92,125.46,60.14,51.36,45.33,44.13,43.72,30.13,25.59,18.26,14.83.ESI-MS m/z:516.2(M+H)⁺,514.1(M-H)^-.

the structural formula of compound 10b is:

example 26

Preparation of compound 10c, isopropyl 3- (6- ((5- ((2-chloro-6-methylphenyl) carbamoyl) -2-thiazolyl) amino) -2-methyl-4-pyrimidinyl) -1-pyrrolidine carboxylate.

The procedure is as in example 1, using compound I-8 to react with isopropyl chloroformate. This gave 10c as a brown solid in 65% yield.

Characterization data for this compound 10c were:¹H NMR(400MHz,DMSO-d₆)δ11.44(s,1H),9.90(s,1H),8.23(s,1H),7.38(s,1H),7.27(s,2H),5.93(s,1H),4.76(s,1H),4.35(s,1H),3.55(s,2H),3.13(s,3H),2.38(s,3H),2.24(s,3H),2.09(s,1H),1.84(s,1H),1.19(s,6H)；¹³C NMR(125MHz,DMSO-d₆)δ165.34,162.67,162.50,159.93,155.82,153.85,140.92,138.78,133.59,132.45,128.89,128.00,126.89,125.47,67.29,52.06,45.27,44.05,43.70,30.04,25.40,22.06,18.26.ESI-MS m/z:529.2(M+H)⁺,528.2(M-H)^-.

the structural formula of compound 10c is:

example 27

Preparation of compound 10d, allyl 3- (6- ((5- ((2-chloro-6-methylphenyl) carbamoyl) -2-thiazolyl) amino) -2-methyl-4-pyrimidinyl) -1-pyrrolidinecarboxylic acid salt.

The preparation was carried out as in example 1, using compound I-8 and allyl chloroformate. Yellow solid 10d was obtained in 44% yield.

Characterization data for this compound 10d were:¹H NMR(400MHz,DMSO-d₆)δ11.46(s,1H),9.93(s,1H),8.25(s,1H),7.39(d,J＝7.0Hz,1H),7.34-7.18(m,2H),5.95(s,1H),5.93-5.80(m,1H),5.29(dd,J＝23.9,8.8Hz,1H),5.17(t,J＝8.3Hz,1H),4.52(s,2H),3.59(s,1H),3.21(s,2H),2.38(s,3H),2.24(s,3H),2.10(s,1H),1.85(s,1H)；¹³C NMR(125MHz,DMSO-d₆)δ165.37,162.69,162.49,159.95,155.83,153.80,140.91,138.80,133.59,132.45,129.00,128.05,126.93,126.17,125.48,116.78,64.73,52.07,45.34,44.26,43.75,30.13,25.42,18.27.ESI-MS m/z:527.2(M+H)⁺,526.2(M-H)^-.

the structural formula of compound 10d is:

example 28

Preparation of compound 10e, butyl 3- (6- ((5- ((2-chloro-6-methylphenyl) carbamoyl) -2-thiazolyl) amino) -2-methyl-4-pyrimidinyl) -1-pyrrolidinecarboxylic acid salt.

The procedure is as in example 1, using compound I-8 to react with butyl chloroformate. 10e was obtained as a brown solid in 59% yield.

Characterization data for this compound 10e were:¹H NMR(400MHz,DMSO-d₆)δ11.44(s,1H),9.89(s,1H),8.22(s,1H),7.39(s,1H),7.28(s,2H),5.93(s,1H),4.36(s,1H),3.97(s,2H),3.56(s,2H),3.18(s,2H),2.38(s,3H),2.24(s,3H),2.10(s,1H),1.84(s,1H),1.53(s,2H),1.32(s,2H),0.88(d,J＝5.0Hz,3H)；¹³C NMR(125MHz,DMSO-d₆)δ165.33,162.67,162.49,159.92,156.36,154.24,140.89,138.79,133.60,132.45,129.40,128.90,127.98,126.89,125.49,64.02,52.08,45.32,44.09,43.67,30.68,30.09,25.40,18.56,18.25,13.52.ESI-MS m/z:544.2(M+H)⁺,542.2(M-H)^-.

the structural formula of compound 10e is:

example 29

Preparation of compound 10f, isobutyl 3- (6- ((5- ((2-chloro-6-methylphenyl) carbamoyl) -2-thiazolyl) amino) -2-methyl-4-pyrimidinyl) -1-pyrrolidine carboxylate.

The procedure is as in example 1, using compound I-8 to react with isobutyl chloroformate. Yellow solid 10f was obtained in 58% yield.

Characterization data for this compound 10f were:¹H NMR(400MHz,DMSO-d₆)δ11.44(s,1H),9.87(s,1H),8.21(s,1H),7.40(d,J＝7.1Hz,1H),7.26(dd,J＝18.0,6.8Hz,2H),5.93(s,1H),5.75(s,1H),4.38(s,1H),3.76(d,J＝6.3Hz,2H),3.65-3.52(m,1H),3.44(d,J＝12.9Hz,1H),3.19(s,1H),2.38(s,3H),2.24(s,3H),2.17-2.04(m,1H),1.85(dd,J＝12.1,5.9Hz,2H),0.88(dd,J＝10.5,6.8Hz,6H)；¹³C NMR(125MHz,DMSO-d₆)δ165.39,162.72,162.51,159.92,155.80,154.21,140.81,138.78,133.53,132.42,128.94,128.06,126.93,125.44,70.24,54.82,45.64,44.11,43.68,30.12,27.54,25.41,18.82,18.24.ESI-MS m/z:544.2(M+H)⁺,542.2(M-H)^-.

the compound 10f has the structural formula:

example 30

Preparation of Compound 10g, phenyl 3- (6- ((5- ((2-chloro-6-methylphenyl) carbamoyl) -2-thiazolyl) amino) -2-methyl-4-pyrimidinyl) -1-pyrrolidinecarboxylic acid salt.

The preparation was carried out as in example 1, using compound I-8 and phenyl chloroformate. 10g of a yellow solid are obtained in 76% yield.

The characterization data for 10g of this compound is:¹H NMR(400MHz,DMSO-d₆)δ11.47(s,1H),10.07(s,1H),9.91(s,1H),8.24(s,1H),7.40-7.35(m,3H),7.28-7.21(m,3H),7.15-7.12(m,2H),5.97(s,1H),4.46(s,1H),3.87-3.75(m,1H),3.72-3.58(m,2H),3.55-3.46(m,1H),2.40(s,3H),2.24(s,3H),2.21-2.12(m,1H),2.01-1.90(m,1H)；¹³C NMR(125MHz,DMSO-d₆)δ165.90,163.21,163.05,163.03,160.44,152.77,151.65,141.43,139.33,134.13,132.97,129.69,129.40,128.56,127.43,125.54,122.28,60.78,52.16,45.81,45.02,44.89,31.49,25.95,18.89.ESI-MS m/z:563.2(M+H)⁺,562.1(M-H)^-.

the compound 10g has the structural formula:

example 31

Preparation of compound 10h, benzyl 3- (6- ((5- ((2-chloro-6-methylphenyl) carbamoyl) -2-thiazolyl) amino) -2-methyl-4-pyrimidinyl) -1-pyrrolidine carboxylate.

The procedure is as in example 1, using compound I-9 to react with benzyl chloroformate. A yellow solid was obtained in 10h, 66% yield.

Characterization data for this compound 10h are:¹H NMR(400MHz,DMSO-d₆)δ11.45(s,1H),9.90(s,1H),8.23(s,1H),7.51-7.28(m,8H),5.83(s,1H),5.07(s,2H),4.40(s,1H),3.61(s,1H),3.45(s,2H),3.26-3.17(m,2H),2.38(s,3H),2.24(s,3H),2.12(s,1H),1.86(s,1H)；¹³C NMR(125MHz,DMSO-d₆)δ165.33,162.81,162.66,162.47,159.91,153.92,140.87,138.74,137.06,133.56,132.43,128.87,128.29,128.00,127.64,127.38,126.89,125.44,65.68,52.05,45.27,44.29,43.79,30.08,25.42,18.25.ESI-MS m/z:578.2(M+H)⁺,576.2(M-H)^-.

the compound 10h has the structural formula:

example 32

Preparation of compound 10i, N- (2-chloro-6-methylphenyl) -2- ((2-methyl-6- ((1- (methylsulfonyl) -3-pyrrolidinyl) amino) 4-pyrimidinyl) amino) -5-thiazolecarboxamide.

Compound I-8 was selected for reaction with methanesulfonyl chloride and synthesized according to the following route:

compound I-8(0.5mmol,1eq), TEA (triethylamine, 0.6mmol, l.2eq), and THF 5mL were added to a 25mL single vial at 0 ℃, methanesulfonyl chloride (0.6mmol, l.2eq) was slowly added, and after the addition, the reaction was carried out at 0 ℃ for 1h, and the reaction was carried out at room temperature. TLC monitoring, reaction is finished, and solvent is removed by rotary evaporation. The solid was washed with a little water and passed through the column to give the product 10i. yellow solid in 91% yield.

Characterization data for this compound 10i are:¹H NMR(400MHz,DMSO-d₆)δ11.47(s,1H),9.90(s,1H),8.23(s,1H),7.39(d,J＝5.1Hz,1H),7.28(s,2H),5.76(s,1H),4.40(s,1H),3.54(s,1H),2.90(s,3H),2.39(s,3H),2.31(s,3H),2.24(s,3H),1.89(s,1H)；¹³C NMR(125MHz,DMSO-d₆)δ165.39,162.64,162.44,159.92,155.81,140.89,138.78,133.56,132.43,128.92,128.04,126.91,125.49,54.84,52.82,45.99,45.40,33.48,30.85,25.42,18.25.ESI-MS m/z:522.1(M+H)⁺,520.1(M-H)^-.

the structural formula of the compound 10i is:

example 33

Preparation of compound 10j, N- (2-chloro-6-methylphenyl) -2- ((2-methyl-6- ((1- (ethanesulfonyl) -3-pyrrolidinyl) amino) 4-pyrimidinyl) amino) -5-thiazolecarboxamide.

Compound I-8 was selected for reaction with ethanesulfonyl chloride and prepared according to the method of example 23. This gave 10j as a brown solid in 88% yield.

Characterization data for this compound 10j were:¹H NMR(400MHz,DMSO-d₆)δ11.49(s,1H),9.92(s,1H),8.27(s,1H),7.36(d,J＝5.1Hz,1H),7.27(s,2H),5.79(s,1H),4.40(s,1H),3.67-3.60(m,2H),3.55(s,1H),2.40(s,3H),2.32(s,3H),2.24(s,3H),1.88(s,1H),0.98(t,J＝4.8Hz,3H)；¹³C NMR(125MHz,DMSO-d₆)δ164.39,162.64,162.47,159.62,155.81,140.59,138.54,133.78,132.63,128.92,127.04,126.96,125.76,55.87,52.92,45.99,45.38,32.57,31.95,24.92,18.29,9.75.ESI-MS m/z:535.1(M+H)⁺,534.1(M-H)^-.

the structural formula of the compound 10j is:

example 34

Compound 10k, (E) -N- (2-chloro-6-methylphenyl) -2- ((6- ((1-acryloyl-3-pyrrolidinyl) amino) -2-methyl-4-pyrimidinyl) amino) -5-thiazolecarboxamide.

The desired product was obtained by a similar synthetic procedure to that of compound 8l in example 12, using compound I-8 to react with acryloyl chloride to give 10k, 85mg, 73% yield as a white solid.

Characterization data for this compound 10k is:¹H NMR(400MHz,DMSO)δ11.44(s,1H),9.87(s,1H),8.22(s,1H),7.50(dd,J＝18.2,6.2Hz,1H),7.40(d,J＝7.1Hz,1H),7.32-7.21(m,2H),6.57(ddd,J＝22.0,16.7,10.3Hz,1H),6.13(dd,J＝16.0,6.2Hz,1H),5.94(s,1H),5.66(t,J＝11.4Hz,1H),3.89-3.59(m,2H),2.87(s,2H),2.39(s,3H),2.24(s,3H),2.09-2.08(m,1H),1.96-1.85(m,2H).¹³C NMR(125MHz,DMSO-d₆)δ165.43,163.43,162.71,162.54,159.96,140.91,138.81,133.58,132.46,129.67,129.30,128.96,128.08,126.95,126.67,125.58,52.04,45.43,44.31,43.78,25.44,18.29.MS(ESI)m/z:499.0(M+H)⁺,497.0(M-H)^-.

the structural formula of compound 10k is:

example 35

The compound 10l, (E) -N- (2-chloro-6-methylphenyl) -2- ((6- ((1- (2-butenoyl) -3-pyrrolidinyl) amino) -2-methyl-4-pyrimidinyl) amino) -5-thiazolecarboxamide.

Referring to a synthetic procedure similar to that for compound 8l in example 12, the desired product was obtained by reacting compound I-8 with 2-butenoyl chloride to give 10l of a white solid, 122mg, in 68.3% yield.

Characterization data for this compound 10l were:¹H NMR(400MHz,DMSO-d₆)δppm 11.44(s,1H),9.87(s,1H),8.21(s,1H),7.48(dd,J＝20.4,5.6Hz,1H),7.40(d,J＝7.1Hz,1H),7.32-7.21(m,2H),6.75-6.61(m,1H),6.35-6.16(m,1H),5.93(s,1H),3.87-3.73(m,1H),3.69-3.47(m,2H),3.02(dd,J＝12.7,6.0Hz,3H),2.39(s,3H),2.24(s,3H),1.87-1.79(m,3H)；¹³C NMR(125MHz,DMSO-d₆)δ165.40,163.68,162.72,162.55,162.48,159.96,140.88,139.74,139.68,138.81,133.57,132.45,128.96,128.08,126.95,125.48,123.66,123.32,62.60,61.99,52.04,18.29,17.61,7.16.MS(ESI)m/z:513.1(M+H)⁺,511.0(M-H)^-.

the structural formula of the compound 10l is:

example 36

Compound 10m, (E) -N- (2-chloro-6-methylphenyl) -2- ((6- ((1-propionyl-3-pyrrolidinyl) amino) -2-methyl-4-pyrimidinyl) amino) -5-thiazolecarboxamide.

Referring to a synthetic procedure similar to compound 8n to obtain the desired product, compound I-8 was selected for reaction with propionic acid to give a white solid 10m, 96mg, 49.2% yield.

Characterization data for this compound 10m is:¹H NMR(400MHz,DMSO-d₆)δ11.42(s,1H),9.88(s,1H),8.20(s,1H),7.39-7.23(m,3H),5.91(s,1H),4.47(s,1H),4.09-4.06(m,1H),3.41-2.98(m,2H),2.38(s,3H),2.27(s,3H),2.09-2.04(m,4H),1.90-1.82(m,2H),0.99-0.95(m,3H).¹³C NMR(125MHz,DMSO-d₆)δ171.64,171.54,165.54,162.69,160.10,155.95,155.35,140.90,138.88,133,59,132.51,129.03,128.18,127.00,125.52,78.96,58.95,54.54,31.30,29.68,26.62,18.26,9.85.MS(ESI)m/z:501.0(M+H)⁺,499.0(M-H)^-.

the structural formula of the compound 10m is:

example 37

First, DDR1 and DDR2 in vitro kinase data for all compounds described above were obtained from FRET (Fluorescence Resonance Energy Transfer) based LANCE Ultra protein kinase assay. The positive control substance is Dasatinib synthesized by the user (the Dasatinib is determined by mass spectrum, infrared and nuclear magnetic resonance hydrogen spectrum and carbon spectrum verification). The specific detection method comprises the following steps:

reagents and materials: the kinases DDR1 and DDR2 were purchased from Carna Bioscience, and the substrate Poly GAT and antibody Eu-PY20antibody were purchased from PE. The 384 microwell plates were purchased from corning, inc, model 3573. The multifunctional microplate reader was purchased from Envision MultilabelReader, PE. The reaction mixture was prepared according to the description of the LANCE Ultra protein kinase assay. The reaction buffer composition was 50mM Tris-HCl (pH 7.4), 0.9% NaCl, 0.1% BSA, 0.05% sodium azide.

And (3) experimental operation: firstly, adding 5 mul of mixed solution of enzyme and ATP into a microporous plate, then adding a series of compounds diluted in a gradient manner by using Echo520, shaking and uniformly mixing for 10min, finally adding 5 mul of substrate into each hole, shaking and uniformly mixing for 5min, and reacting for 1.5h at room temperature; then 10 mul of antibody detection solution with corresponding concentration is added, the mixture is shaken and mixed evenly for 5min, and the detection is carried out after the reaction is carried out for 1h at room temperature. (excitation light wavelength 3400nm, emission light wavelength 665nm, respectively).

Finally calculating the IC of the compound₅₀: EC is obtained by analyzing the concentration of the compound and the fluorescence value obtained by detection on GraphPad software₅₀Value i.e. IC of the compound₅₀The value is obtained.

Secondly, the kinase inhibition activity of Abl, Src, Btk, Kit and the like of the compound is measured by adopting Z' -LYTE^TMThe kinase test kit performs the test. The positive control was self-synthesized Dasatinib. The method specifically comprises the following steps:

reagents and materials: the kinases Abl, Src, Btk and Kit used were purchased from Life technologies; the substrates Tyr2, Tyr1 and Tyr6 kits (the substrate kit comprises a substrate, a detection solution, a stop solution and a reaction buffer solution) are purchased from Life technologies; 384 microplates were purchased from NUC corporation under model 267461; the micro liquid handling system Echo520 was purchased from labctye; the multifunctional microplate reader was purchased from Envision MultilabelReader, PE. The reaction solution comprises the following components: appropriate amounts of enzyme, 2 μ M substrate, corresponding concentrations of ATP and a series of serially graded dilutions of the compound. The reaction buffer composition was 50mM HEPES pH 7.5,10mM MgCl₂,1mM EGTA，0.01％BRIJ-35。

And (3) experimental operation: firstly, adding 5ul of a proper amount of enzyme and corresponding substrate (2 mu M) mixed solution into a microporous plate, then adding a series of compounds diluted in a gradient manner by using Echo520, shaking and uniformly mixing for 10min, finally adding ATP (adenosine triphosphate) by using Echo, shaking and uniformly mixing for 5min, and placing in a constant temperature box with 29 ℃ for reacting for 1.5 h; then adding 2.5 μ l of detection solution (development reagent) with corresponding concentration, shaking and mixing for 5min, and placing in a constant temperature box of 29 ℃ for reaction for 1 h; finally, 7.5. mu.l of stop solution (StopReagent) is added, and the mixture is shaken and mixed uniformly for detection. (excitation light wavelength 400nm, emission light wavelength 445nm and 520nm, respectively).

The assay was set up with test wells (plus compound, enzyme, substrate and ATP), 0% phosphorylated wells (plus DMSO, substrate and ATP) 100% phosphorylated wells (plus phosphorylated substrate only), 0% inhibitory wells (plus DMSO, enzyme, substrate and ATP), respectively.

Then calculating the phosphorylation rate of each well according to a formula, and finally analyzing the EC on GraphPad software by using the compound concentration and the calculated activity rate of the enzyme₅₀Value i.e. IC of the compound₅₀The value is obtained.

And thirdly, testing the proliferation inhibition activity of the K562 cells by adopting a CCK-8 detection method.

Experimental materials: k562 cells, medium RPMI1640 (10% FBS, 1% PS); corning cell culture plate (#3701)

An experimental instrument: multi-label microplate detector (model Envision multilabel reader).

And (3) experimental operation: tumor cells in the logarithmic growth phase of cells were seeded into 384-well plates (500 cells/50 μ L/well). A DMSO solution of compound at a concentration of 10mM was prepared and diluted from 10. mu.M to 0.5nM for at least 3 concentration gradients. After mixing the blank control group and the vehicle control group with the drug to be tested, incubating for 72H, and adding 3 μ L (5mg/ml) of CCK-8 reagent (containing WST-8, i.e. 2- (2-methoxy-4-nitrophenyl) -3- (4-nitrophenyl) -5- (2, 4-disulfonate-phenyl) -2H-tetrazole monosodium salt) to each well. After further incubation at 37 ℃ for 2 hours, the absorbance (OD) at the wavelengths of 450nM and 650nM was measured using a microplate reader₄₅₀And OD₆₅₀). Wherein, OD₄₅₀Is the absorbance value, OD, of the reaction product of the CCK-8 colorimetry₆₅₀Is the absorbance value of CCK-8 reagent. The actual absorbance value OD (OD ═ OD) of each well obtained by calculation₄₅₀-OD₆₅₀) The cell viability was calculated according to the following formula:

wherein, OD_experimentIs the absorbance value of the test well (containing cells and different concentrations of the drug to be tested), OD_controlAbsorbance value of 0% inhibition pore, OD_blankAbsorbance values for blank control wells.

Finally calculating the IC of the compound₅₀: EC was determined by analysis on GraphPad software using compound concentrations and calculated viability of cells₅₀Value i.e. IC of the compound₅₀The value is obtained.

The results are shown in the following table.

TABLE 1 half Inhibitory Concentration (IC) of each compound on kinase such as DDR1, DDR2, Abl, Src, Btk and Kit, K562 cells_50，nM)

From the results, the tyrosine kinase small molecule inhibitor provided by the invention has an inhibiting effect on kinases such as DDR1, DDR2, Abl, Src, Btk and Kit, and the like, and the half inhibitory concentration of the tyrosine kinase small molecule inhibitor is superior to or at least equal to that of a positive control dasatinib (dasatinib), especially the compound 8j, and the tyrosine kinase small molecule inhibitor has good enzyme inhibitory activity and cell activity on five tyrosine kinases such as DDR1, DDR2, Src, Btk and Kit, and the half inhibitory concentration of the tyrosine kinase small molecule inhibitor on K562 cells is smaller than that of the positive control.

The technical features of the embodiments described above may be arbitrarily combined, and for the sake of brevity, all possible combinations of the technical features in the embodiments described above are not described, but should be considered as being within the scope of the present specification as long as there is no contradiction between the combinations of the technical features.

The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.

Claims (10)

1. A tyrosine kinase inhibitor having the structural features of formula I:

wherein:

R₁，R₂independently selected from: C1-C6 alkyl, halogen substituted C1-C6 alkyl, C1-C6 alkoxy, halogen substituted C1-C6 alkoxy, C2-C6 alkenyl, halogen substituted C2-C6 alkenyl, C2-C6 alkynyl, halogen substituted C2-C6 alkynyl, C3-C6 cycloalkyl, halogen substituted C3-C6 cycloalkyl, a three-to six-membered heterocycle containing N, O, S heteroatoms, halogen, or H;

the five-membered cyclic group comprising A, B is:

wherein: r₆Independently selected from: h, C1-C6 alkyl, halogen-substituted methyl, OR₇，NR₇R₈，CN，COOR₇，CONR₇R₈，SO₂R₇，SO₂NR₇R₈，NO₂，CSNR₇R₈；

R₇、R₈Independently selected from: H. C1-C6 alkyl, N, S, O heteroatom substituted C1-C6 alkyl;

v is:

wherein: r₉Selected from: h, C1-C6 alkyl, halogen substituted C1-C6 alkyl, C1-C6 alkoxy;

w is selected from one of the following groups:

m is selected from: 1 to 6;

x is selected from: o, S, or none;

R₃selected from: C1-C6 alkyl, halogen substituted C1-C6 alkyl, C1-C6 alkoxy, C1-C6 nitrogen-containing alkyl,

phenyl or one of the following substituents:

wherein: n is selected from: 1 to 6;

R₁₀，R₁₁，R₁₂independently selected from: h, C1-C6 alkyl, halogen substituted C1-C6 alkyl, C1-C6 alkoxy.

2. A tyrosine kinase inhibitor or a pharmaceutically acceptable salt thereof according to claim 1, characterized by being selected from compounds having the following structural features of formula II:

R₁，R₂，R₃v, W, X are as defined in claim 1.

3. A tyrosine kinase inhibitor or a pharmaceutically acceptable salt thereof according to claim 2, characterized in that:

v is:

w is selected from one of the following groups:

x is selected from: o, or none;

R₉as claimed in claim 1.

4. The tyrosine kinase inhibitor or a pharmaceutically acceptable salt thereof according to claim 3, wherein R is₉Selected from: H.

5. the tyrosine kinase inhibitor or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 4, wherein R is₃Selected from: C1-C6 alkyl, halogen substituted C1-C6 alkyl, C1-C6 nitrogen-containing alkyl, phenyl or the following substituents:

wherein: n is selected from: 1 to 6;

R₁₀selected from: H.

6. a tyrosine kinase inhibitor or a pharmaceutically acceptable salt thereof according to claim 5,

x is selected from: o;

R₃selected from: C1-C6 alkyl, halogen substituted C1-C6 alkyl, C1-C6 nitrogenous alkyl, or N-ethylmorpholine.

7. The tyrosine kinase inhibitor or a pharmaceutically acceptable salt or stereoisomer thereof according to claim 1, wherein R is₁，R₂Independently selected from: C1-C6 alkyl, or halogen.

8. A tyrosine kinase inhibitor or a pharmaceutically acceptable salt or stereoisomer thereof according to claim 1, characterized by being selected from one of the following compounds:

3- (6- ((5- ((2-chloro-6-methylphenyl) carbamoyl) -2-thiazolyl) amino) -2-methyl-4-pyrimidinyl) -1-pyrrolidinecarboxylic acid methyl ester;

3- (6- ((5- ((2-chloro-6-methylphenyl) carbamoyl) -2-thiazolyl) amino) -2-methyl-4-pyrimidinyl) -1-pyrrolidinecarboxylic acid isopropyl ester;

3- (6- ((5- ((2-chloro-6-methylphenyl) carbamoyl) -2-thiazolyl) amino) -2-methyl-4-pyrimidinyl) -1-pyrrolidinecarboxylic acid allyl ester;

3- (6- ((5- ((2-chloro-6-methylphenyl) carbamoyl) -2-thiazolyl) amino) -2-methyl-4-pyrimidinyl) -1-pyrrolidinecarboxylic acid butyl ester;

3- (6- ((5- ((2-chloro-6-methylphenyl) carbamoyl) -2-thiazolyl) amino) -2-methyl-4-pyrimidinyl) -1-pyrrolidinecarboxylic acid isobutyl ester;

phenyl 3- (6- ((5- ((2-chloro-6-methylphenyl) carbamoyl) -2-thiazolyl) amino) -2-methyl-4-pyrimidinyl) -1-pyrrolidinecarboxylate;

benzyl 3- (6- ((5- ((2-chloro-6-methylphenyl) carbamoyl) -2-thiazolyl) amino) -2-methyl-4-pyrimidinyl) -1-pyrrolidinecarboxylate;

2- ((2-methyl-6- ((1- (methylsulfonyl) -3-pyrrolidinyl) amino) 4-pyrimidinyl) amino) -5-thiazolecarboxamide;

2- ((2-methyl-6- ((1- (ethylsulfonyl) -3-pyrrolidinyl) amino) 4-pyrimidinyl) amino) -2- (2-chloro-6-methylphenyl) -5-thiazolecarboxamide;

(E) -N- (2-chloro-6-methylphenyl) -2- ((6- ((1-acryloyl-3-pyrrolidinyl) amino) -2-methyl-4-pyrimidinyl) amino) -5-thiazolecarboxamide;

(E) -N- (2-chloro-6-methylphenyl) -2- ((6- ((1- (2-butenoyl) -3-pyrrolidinyl) amino) -2-methyl-4-pyrimidinyl) amino) -5-thiazolecarboxamide;

(E) -N- (2-chloro-6-methylphenyl) -2- ((6- ((1-propionyl-3-pyrrolidinyl) amino) -2-methyl-4-pyrimidinyl) amino) -5-thiazolecarboxamide.

9. A process for the preparation of a tyrosine kinase inhibitor or a pharmaceutically acceptable salt thereof as claimed in any one of claims 1 to 8, which is synthesised by the following route:

10. use of the tyrosine kinase inhibitor or the pharmaceutically acceptable salt thereof according to any one of claims 1 to 8 for the preparation of a medicament for the prevention and treatment of an antitumor drug.