CN113727981B

CN113727981B - Kinase inhibitor, preparation, pharmaceutical composition and application thereof

Info

Publication number: CN113727981B
Application number: CN202080030257.5A
Authority: CN
Inventors: 皮士卿; 徐燕; 杨代鸿; 周志刚
Original assignee: Hunan New Medical Technology Development Co ltd; Hunan Warrant Pharmaceutical Technology Development Co ltd; Hunan Warrant Pharmaceutical Co ltd
Current assignee: Hunan New Medical Technology Development Co ltd; Hunan Warrant Pharmaceutical Technology Development Co ltd; Hunan Warrant Pharmaceutical Co ltd
Priority date: 2019-08-20
Filing date: 2020-08-19
Publication date: 2023-01-20
Anticipated expiration: 2040-08-19
Also published as: CN112409349A; WO2021032129A1; CN113727981A

Abstract

The present application relates to compounds of formula I that are useful as kinase inhibitors (e.g., tyrosine kinase inhibitors), methods for their preparation, and pharmaceutical compositions comprising the compounds of formula I. The application also relates to the therapeutic application of the compound of the formula I or the pharmaceutical composition thereof in immune diseases, tumors, neurological diseases and other diseases.

Description

Kinase inhibitor, preparation, pharmaceutical composition and application thereof

Cross Reference to Related Applications

This application claims priority and benefit from the chinese patent application No. 201910771036.2 filed on 20/8/2019 with the chinese intellectual property office, the disclosure of which is incorporated herein by reference in its entirety.

Technical Field

The present application is in the field of pharmaceutical chemistry and in particular relates to compounds which are kinase inhibitors (e.g. tyrosine kinase inhibitors), processes for their preparation and pharmaceutical compositions comprising said compounds. The application also relates to the therapeutic application of the compound or the pharmaceutical composition thereof in immune diseases, tumors, neurological diseases and other diseases.

Background

Protein kinases are a class of enzymes that catalyze the phosphorylation of proteins, thereby altering their substrate activity or their ability to bind to other proteins. The kinase signaling pathway is the most common form of reversible post-translational modification, controlling many aspects of cellular function. Abnormal activation of protein kinases is a major hallmark of malignant tumors, including alterations in cell proliferation, survival, motility, and metabolism, as well as diseases such as vascular proliferation and evasion of anti-tumor immune responses. The new blood vessels that are created "supply" the oxygen and nutrients for growing tumors.

1.BCR-ABL

A large number of researches prove the function of Bcr-Abl tyrosine kinase in the pathogenesis of Chronic Myelogenous Leukemia (CML), and the Bcr-Abl inhibitor paves the way for developing a novel targeted therapeutic small molecule drug. Imatinib (Imatinib), bosutinib (Bosutinib), dasatinib (Dasatinib), nilotinib (Nilotinib), ladostinib (raditinib), and panatinib (Ponatinib) have been approved for the treatment of chronic myelocytic leukemia eosinophilic syndrome, myeloproliferative/myeloproliferative tumors, chronic myelocytic leukemia, acute lymphoblastic leukemia/lymphoblastic lymphoma.

Although these drugs increase the survival rate of tumor patients, they also cause severe adverse reactions, such as severe diarrhea. This problem is particularly pronounced in the stomach and upper digestive tract, as the drug concentration is higher in this area after oral administration. At this drug concentration, poorly selective tyrosinase inhibitors (TKIs) can inhibit protein kinases abundantly expressed in the gastrointestinal tract, resulting in severe common dose-related toxic reactions.

2.BTK

Bruton's Tyrosine Kinase (BTK) is a non-receptor kinase that has been demonstrated to be a genetic factor in X-linked agammaglobulinemia. BTK plays a crucial role in oncogenic signals that play an important role in the proliferation and survival of leukemic cells of many B-cell malignancies. Therefore, BTK is a key target for the development of small molecule inhibitors of B-cell malignancies.

Although Bcr-Abl inhibitors are currently approved for tumor therapy and can fundamentally improve the average survival rate of patients. However, new gene mutations and drug-resistant forms also appear in clinical practice, and thus further development of new inhibitors is required. Large doses of drug can cause mutations in the target protein, resulting in loss of activity. There is still a need to find a low dose Bcr-Abl inhibitor and a reversible BTK inhibitor that reduces side effects and the possibility of mutations to meet the medical need.

Disclosure of Invention

In one aspect, the present application provides a compound of formula I, or a pharmaceutically acceptable salt, ester, solvate, prodrug, active metabolite, crystal, stereoisomer, tautomer, or geometric isomer thereof,

wherein:

R ₀ selected from halogen and D;

R ₂ 、R ₃ 、R ₅ 、R ₇ 、R ₁₀ 、R ₁₁ 、R ₁₂ 、R ₁₃ 、R ₁₄ 、R ₁₅ 、R ₁₆ 、R ₁₇ each independently selected from H and D;

R ₁ 、R ₄ and R ₆ Each independently selected from optionally substituted alkyl, optionally substituted hydroxyalkyl, halogen, optionally substituted alkoxy, optionally substituted alkynyl, optionally substituted alkenyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted carboxyalkyl, -NO ₂ -CN, -OH, -COOH, thiol, optionally substituted amino, optionally substituted alkylthio, optionally substituted ester, optionally substituted acyl, and optionally substituted sulfonyl;

R ₈ selected from the group consisting of H, D, optionally substituted hydroxyalkyl, optionally substituted carboxyalkyl, optionally substituted alkyl, halogen, optionally substituted alkoxy, optionally substituted alkynyl, optionally substituted alkenyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, -NO ₂ -CN, -OH, -COOH, thiol, optionally substituted amino, optionally substituted alkylthio, optionally substituted ester, optionally substituted acyl, and optionally substituted sulfonyl.

In some embodiments, R ₁ 、R ₄ And R ₆ Each independently selected from optionally substituted C _1-6 Alkyl, optionally substituted C _1-6 Hydroxyalkyl, halogen, optionally substituted C _1-6 Alkoxy, optionally substituted C _2-6 Alkynyl, optionally substituted C _2-6 Alkenyl, optionally substituted 3-8 membered cycloalkyl, optionally substituted 3-10 membered heterocyclyl, optionally substituted 6-10 membered aryl, optionally substituted 5-10 membered heteroaryl, optionally substituted C _1-6 Carboxyalkyl, -NO ₂ -CN, -OH, -COOH, mercapto, optionally substituted amino, optionally substituted C _1-6 Alkylthio, optionally substituted ester, optionally substituted acyl and optionally substituted sulfonyl.

In some embodiments, R ₈ Selected from H, D, optionally substituted C _1-6 Hydroxyalkyl, optionally substituted C _1-6 Carboxyalkyl, optionally substituted C _1-6 Alkyl, halogen, optionally substituted C _1-6 Alkoxy, optionally substituted C _2-6 Alkynyl, optionally substituted C _2-6 Alkenyl, optionally substituted 3-to 8-membered cycloalkyl, optionally substituted 3-to 10-membered heterocyclyl, optionally substituted 6-to 10-membered aryl, optionally substituted 5-to 10-membered heteroaryl, -NO ₂ -CN, -OH, -COOH, mercapto, optionally substituted amino, optionally substituted C _1-6 Alkylthio, optionally substituted ester, optionally substituted acyl and optionally substituted sulfonyl.

In some embodiments, one or more heteroatoms in the heterocyclyl and heteroaryl groups are each independently selected from O, S, and N; in some embodiments, the number of heteroatoms is each independently selected from 1, 2, and 3.

In some embodiments, R ₀ Selected from fluorine, chlorine, bromine and iodine. In some embodiments, R ₀ Selected from fluorine.

In some embodiments, R ₂ 、R ₃ 、R ₅ 、R ₇ 、R ₁₀ 、R ₁₁ 、R ₁₂ 、R ₁₃ 、R ₁₄ 、R ₁₅ 、R ₁₆ 、R ₁₇ Each independently selected from H.

In some embodiments, R ₁ 、R ₄ And R ₆ Each independently selected from optionally substituted C _1-6 Alkyl, optionally substituted C _1-6 Hydroxyalkyl, and halogen.

In some embodiments, R ₁ And R ₄ Each independently selected from C _1-6 Alkyl and halogen.

In some embodiments, R ₁ Is selected from C _1-4 Alkyl radical, C _1-4 Haloalkyl, C _1-4 Hydroxyalkyl and halogen.

In some embodiments, R ₁ Is selected from-CH ₃ 、-CH ₂ OH and chlorine.

In some casesIn the embodiment, R ₁ Is selected from-CH ₃ And chlorine.

In some embodiments, R ₄ Is selected from C _1-4 Alkyl radical, C _1-4 Haloalkyl, C _1-4 Deuterated alkyl and halogen.

In some embodiments, R ₄ Is selected from-CH ₃ 、-CD ₃ Fluorine and chlorine.

In some embodiments, R ₄ Is selected from-CH ₃ Fluorine and chlorine.

In some embodiments, R ₄ Is selected from-CH ₃ And chlorine.

In some embodiments, R ₆ Is selected from C _1-4 Alkyl radical, C _1-4 Haloalkyl, C _1-4 Deuterated alkyl, C _1-4 Hydroxyalkyl, and halogen.

In some embodiments, R ₆ Is selected from C _1-4 An alkyl group.

In some embodiments, R ₆ Is selected from-CH ₃ 、-CH ₂ OH and-CD ₃ 。

In some embodiments, R ₆ Is selected from-CH ₃ 。

In some embodiments, R ₈ Selected from H, D, optionally substituted C _1-6 Hydroxyalkyl and optionally substituted C _1-6 A carboxyalkyl group.

In some embodiments, R ₈ Selected from H, D, C _1-4 Hydroxyalkyl and C _1-4 A carboxyalkyl group.

In some embodiments, R ₈ Is selected from C _1-4 A hydroxyalkyl group.

In some embodiments, R ₈ Selected from H, D, -CH ₂ CH ₂ OH and-CH ₂ CO ₂ H。

In some embodiments, R ₈ Is selected from-CH ₂ CH ₂ OH。

In some embodiments, the term "optionally substituted" refers to a group that is unsubstituted or substituted with one or more substituents independently selected from: the amount of deuterium is such that the deuterium is,halogen, hydroxy, C ₁ -C ₆ Alkyl radical, C ₂ -C ₆ Alkenyl radical, C ₂ -C ₆ Alkynyl, C _1-6 Hydroxyalkyl, C _1-6 Alkoxy, 3-to 8-membered cycloalkyl, 3-to 10-membered heterocyclyl, 6-to 10-membered aryl, 5-to 10-membered heteroaryl, -NO ₂ CN, -OH, -COOH, mercapto and amino.

In another aspect, the present application provides a compound, or a pharmaceutically acceptable salt, ester, solvate, prodrug, active metabolite, crystal, stereoisomer, tautomer, or geometric isomer thereof,

in yet another aspect, the present application provides a pharmaceutical composition comprising at least any one of the compounds provided in the present disclosure, or a pharmaceutically acceptable salt, ester, solvate, prodrug, active metabolite, crystal, stereoisomer, tautomer, or geometric isomer thereof. In some embodiments, the compositions provided herein further comprise a pharmaceutically acceptable carrier, excipient, or adjuvant. In some embodiments, the pharmaceutical composition of the present disclosure further comprises another therapeutic agent for combination therapy. In some embodiments, the additional therapeutic agent is selected from one or more of the following: cyclophosphamide, ifosfamide, vincristine, daunorubicin, doxorubicin, cytarabine, mitoxantrone, dacarbazine, idarubicin, tretinoin, prednisone, dexamethasone, mercaptopurine, methotrexate, paclitaxel, melphalan, long-acting interferon, venetock, crizotinib, erlotinib, ocitinib, ruxotinib, afatinib, erlotinib, imatinib, lapatinib, bevacizumab, trastuzumab, rituximab, cetuximab, bornauzumab, fludarabine, gemcitabine, decitabine, capecitabine, bendamustine, everolimus, temsirolimus, etoposide, granulocyte colony stimulating factor, temozolomide, zoledram, olixaplatin, cisplatin, carboplatin, and fulvestrant.

In yet another aspect, the present application provides a use of a compound described in the present disclosure, or a pharmaceutically acceptable salt, ester, solvate, prodrug, active metabolite, crystal, stereoisomer, tautomer, or geometric isomer thereof, or a pharmaceutical composition thereof for the preparation of a medicament for the prevention or treatment of a tyrosine kinase-associated disease.

In a further aspect, the present application provides the use of a compound described in the present disclosure, or a pharmaceutically acceptable salt, ester, solvate, prodrug, active metabolite, crystal, stereoisomer, tautomer, or geometric isomer thereof, or a pharmaceutical composition thereof, for the manufacture of a medicament for inhibiting tyrosine kinase activity.

In another aspect, the present application provides a method of preventing or treating a tyrosine kinase-associated disease or disorder, comprising administering to a subject in need thereof a therapeutically effective amount of at least any one of the compounds provided by the present disclosure, or a pharmaceutically acceptable salt, ester, solvate, prodrug, active metabolite, crystal, stereoisomer, tautomer, or geometric isomer thereof, or a pharmaceutical composition thereof.

In yet another aspect, the present application provides a method of inhibiting tyrosine kinase activity, comprising administering to a subject or a tissue or cell thereof at least any one of the compounds provided by the present disclosure or a pharmaceutically acceptable salt, ester, solvate, prodrug, active metabolite, crystal, stereoisomer, tautomer, or geometric isomer thereof, or a pharmaceutical composition thereof. In some embodiments, the method of inhibiting tyrosine kinase activity is performed in vitro or in vivo. In some embodiments, the method of inhibiting tyrosine kinase activity comprises an in vitro or in vivo assay.

In yet another aspect, the present application provides a compound described in the present disclosure, or a pharmaceutically acceptable salt, ester, solvate, prodrug, active metabolite, crystal, stereoisomer, tautomer, or geometric isomer thereof, or a pharmaceutical composition thereof, for use in inhibiting tyrosine kinase activity.

In another aspect, the present application provides a compound described in the present disclosure, or a pharmaceutically acceptable salt, ester, solvate, prodrug, active metabolite, crystal, stereoisomer, tautomer, or geometric isomer thereof, or a pharmaceutical composition thereof, for use in the prevention or treatment of a tyrosine kinase-associated disease.

In some embodiments, the tyrosine kinase-associated diseases are diseases, disorders, and conditions that benefit from inhibition or reduction of tyrosine kinase activity.

In some embodiments, the tyrosine kinase comprises a Bcr-Abl tyrosine kinase and a BTK tyrosine kinase.

In some embodiments, the disease is selected from cancer.

In some embodiments, the cancer is selected from: chronic Myelogenous Leukemia (CML), gastrointestinal stromal tumor (GIST), small Cell Lung Cancer (SCLC), non-small cell lung cancer (NSCLC), multiple myeloma, solid tumor, B-cell lymphoma, chronic Lymphocytic Leukemia (CLL), acute Lymphocytic Leukemia (ALL), non-hodgkin's lymphoma (NHL), small Lymphocytic Lymphoma (SLL), mantle Cell Lymphoma (MCL), melanoma, mastocytosis, germ cell tumor, acute Myeloid Leukemia (AML), marginal zone/diffuse large B cell lymphoma, sarcoma, pancreatic cancer, glioblastoma, head and neck tumors, macroglobulinemia, follicular central lymphoma, prostate cancer, myelodysplastic syndrome, atherosclerotic myelohyperplasia, myelofibrosis, eosinophilia, polycythemia vera, liver cancer, advanced sarcoma, glioblastoma multiforme, gliosarcoma, malignant mesothelioma, melanoma, head and neck cell carcinoma, skin carcinoma, neuroendocrine tumors, B cell lymphoma, gastric cell lymphoma, acute lymphoblastic leukemia, metastatic lymphoma of small cell lymphoma, metastatic carcinoma of the rectum, metastatic carcinoma of the head and neck, metastatic carcinoma of the bladder, metastatic carcinoma of small cell lymphoma, metastatic carcinoma of the prostate and neck.

In some embodiments, the cancer is selected from the group consisting of cancers with chemotherapeutic agents resistant to target BCR-ABL and c-KIT, and cancers with resistance to imatinib.

In some embodiments, the diseases, disorders, and conditions are selected from: bone metastasis, hypercalcemia and/or osteoporosis; pulmonary fiber disease; cardiovascular disease or condition; mast cell mediated inflammatory diseases; HTLV-1 associated myelopathy/tropical spastic paralysis; complex Regional Pain Syndrome (CRPS); weight loss or fat loss; arterial occlusive disease; ubiquitination; diseases or conditions associated with reduced function of degrading sugars; fridriich ataxia; parkinson's disease progression graft rejection; rheumatoid arthritis; graft versus host disease; (ii) an autoimmune disease; recurrent immune thrombocytopenic purpura, pemphigus vulgaris, systemic lupus erythematosus, scleroderma interstitial pulmonary fibrosis and idiopathic urticaria.

In some embodiments, the cardiovascular disease or condition is a cardiovascular disease caused by RASopathy, or a congenital heart disease associated with nosan or a nosan syndrome.

In some embodiments, the mast cell mediated inflammatory disease is selected from osteoarthritis, asthma, chronic obstructive pulmonary disease, uveitis, aspirin-aggravated respiratory disease (AERD), and parkinson's disease.

Detailed Description

Definition of

The following definitions and methods are provided to better define the present application and to guide those of ordinary skill in the art in the practice of the present application. Unless otherwise noted, terms are to be understood according to conventional usage by those of ordinary skill in the relevant art. All patent documents, academic papers, and other publications cited herein are incorporated by reference in their entirety.

The terms "optionally" or "optionally" mean that the subsequently described event or circumstance may or may not occur, and that the description includes instances where said event or circumstance occurs and instances where it does not.

Numerical ranges indicating the number of carbon atoms are used herein to refer to the ranges givenEach integer of (1), e.g. "C ₁ -C ₁₀ 'OR' C _1-10 By "is meant that the group may have 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 carbon atoms.

The term "element" is intended to mean the number of backbone atoms constituting a ring. For example, "3-to 10-membered" means that the number of backbone atoms constituting the ring is 3, 4, 5, 6, 7, 8, 9 or 10.

The term "alkyl" refers to a straight or branched chain saturated hydrocarbon group, which may be monovalent (e.g., methyl), divalent (e.g., methylene), or multivalent (e.g., methine). Examples of the alkyl group include methyl (Me), ethyl (Et), propyl (e.g., n-propyl and isopropyl), butyl (e.g., n-butyl, isobutyl, s-butyl, t-butyl), pentyl (e.g., n-pentyl, isopentyl, neopentyl) and the like; for example, the term "C _1-6 Alkyl "refers to an alkyl group containing 1 to 6 (e.g., 1, 2, 3, 4, 5, 6) carbon atoms (e.g., methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, 1-methylbutyl, 2-methylbutyl, 3-methylbutyl, neopentyl, hexyl, 2-methylpentyl, and the like).

The term "hydroxyalkyl" refers to-alkyl-OH, wherein "alkyl" is as defined above and "optionally substituted hydroxyalkyl" refers to a group in which the alkyl moiety is substituted or unsubstituted.

The term "halo" or "halogen" by itself or as part of another substituent means a fluorine (F), chlorine (Cl), bromine (Br), or iodine (I) atom.

The term "alkoxy" refers to-O-alkyl, wherein "alkyl" is as defined above, and examples of "alkoxy" include, but are not limited to, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, sec-butoxy, tert-butoxy, n-pentoxy, isopentoxy. "optionally substituted alkoxy" means that the alkyl portion of the group is substituted or unsubstituted.

The term "C _2-6 Alkenyl "means having 2 to 6 carbon atoms and preferably 2 to 4 carbon atoms and having at least 1, preferably 1 to 2 sites of unsaturation (>C＝C<) A straight or branched hydrocarbon group of (1). Such groups are exemplified by vinyl, allyl and but-3-en-1-yl.The term includes cis and trans isomers or mixtures of such isomers.

The term "C _2-6 Alkynyl "refers to a straight or branched chain monovalent hydrocarbon radical having from 2 to 6 carbon atoms and preferably from 2 to 3 carbon atoms and having at least 1, preferably from 1 to 2 sites of acetylenic (-C ≡ C-) unsaturation. Examples of such alkynyl groups include ethynyl (-C.ident.CH) and propargyl (-CH) ₂ C≡CH)。

The term "cycloalkyl" refers to a saturated, non-aromatic cyclic hydrocarbon group consisting of carbon and hydrogen atoms, preferably containing 1 or 2 rings. The cycloalkyl group may be a monocyclic, fused polycyclic, bridged, or spiro ring structure. Non-limiting examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, bicyclo [2.2.1] heptyl, and spiro [3.3] heptyl and the like.

The term "heterocyclyl" refers to a saturated or unsaturated, nonaromatic cyclic group containing one or more heteroatoms (e.g., O, N, S) as part of the ring structure and having from 2 to 9 ring carbon atoms; "optionally substituted heterocyclyl" refers to an unsubstituted heterocyclyl or a heterocyclyl substituted with one or more substituents as defined below.

The term "aryl" means an aromatic ring in which each of the ring-forming atoms is carbon. The aryl ring may be composed of five, six, seven, eight, nine or more than nine atoms. Examples of aryl groups include, but are not limited to, phenyl, naphthyl, phenanthryl, anthracyl, fluorenyl, and indenyl. Depending on the structure, the aryl group may be a monovalent group or a divalent group, i.e., an arylene group.

The term "heteroaryl" means a stable 5, 6, 7 membered monocyclic or bicyclic or 7, 8, 9 or 10 membered bicyclic heterocyclic aromatic ring containing carbon atoms and 1, 2, 3 or 4 heteroatoms independently selected from N, O and S. The nitrogen atom may be optionally substituted (i.e. N or NR, wherein R is H or other substituents defined below). The nitrogen and sulfur atoms may optionally be oxidized (i.e., NO and S (O) _p And p is 1 or 2). Non-limiting examples of monocyclic heteroaryl groups include pyridyl; non-limiting examples of fused ring heteroaryl groups include benzimidazolyl (benzimidazolyl), quinolyl (quinolinyl), acridinyl (acridininyl)l), non-limiting examples of non-fused bis-heteroaryl groups include bipyridyl (bipyridinyl). Other examples of heteroaryl groups include, but are not limited to: furyl (furyl), thienyl (thienyl), oxazolyl (oxazolyl), acridinyl (acridinyl), phenazinyl (phenazinyl), isoxazolyl (isoxazoyl), isothiazolyl (isothiazolyl), pyrazolyl (pyrazolyl), purinyl (purinyl), quinolyl (quinolinyl), triazolyl (triazolyl), thiazolyl (thiazolyl), triazinyl (triazonyl), and thiadiazolyl (thiadiazolyl), and the like, and oxides thereof, such as pyridyl-N-oxide (pyridyl-N-oxide).

The term "carboxyalkyl" refers to a group having the structure-alkyl-COOH, wherein alkyl is as defined above. Representative examples of carboxyalkyl groups include, but are not limited to, carboxymethyl (-CH) ₂ CO ₂ H) 2-carboxyethyl and the like. "optionally substituted carboxyalkyl" means that the alkyl portion of the group is substituted or unsubstituted.

The term "optionally substituted amino" refers to-NH ₂ Mono-or di-substituted amino, and 5-to 7-membered cyclic amino.

The term "mercapto" or "thiol" refers to the group-SH.

The term "alkylthio" refers to the group-S-alkyl, wherein alkyl is as defined herein. "optionally substituted alkylthio" means that the alkyl moiety in the group is substituted or unsubstituted.

The term "ester group" as used herein refers to a group having the structure-C (O) OR ', wherein R' is optionally substituted alkyl, optionally substituted aryl OR optionally substituted heteroaryl as defined above. In some embodiments, R' is alkyl, haloalkyl, aryl, heteroaryl, alkaryl, alkylheteroaryl. In some embodiments, R' is alkyl or haloalkyl. In some embodiments, R' is alkyl. By "optionally substituted ester group" is meant that the R' moiety in the group is substituted or unsubstituted.

The term "acyl" as used herein refers to a group having the structure-C (O) R ', wherein R' is as defined above. "optionally substituted acyl" means that the R' moiety in the group is substituted or unsubstituted.

The term "sulfonyl" as used herein means having an-SO ₂ A group of the structure R ', wherein R' is as defined above. "optionally substituted sulfonyl" means that the R' moiety in the group is substituted or unsubstituted.

The term "optionally substituted" means that the group is unsubstituted or substituted with one or more substituents independently selected from: deuterium, halogen, hydroxy, C ₁ -C ₆ Alkyl radical, C ₂ -C ₆ Alkenyl radical, C ₂ -C ₆ Alkynyl, C _1-6 Hydroxyalkyl, C _1-6 Alkoxy, 3-to 8-membered cycloalkyl, 3-to 10-membered heterocyclyl, 6-to 10-membered aryl, 5-to 10-membered heteroaryl, -NO ₂ -CN, -OH, -COOH, mercapto and amino. When a group is substituted with more than one substituent, the substituents may be the same or different, any substituted functional group herein may be substituted at 1 to 3 different positions, and those 1 to 3 substituent groups can each be independently substituted at 1 to 3 positions.

The term "pharmaceutically acceptable salt" refers to salts that retain the biological potency of the free acid and free base of the specified compound, and that are biologically or otherwise not adversely affected. The compounds described herein can have acidic or basic groups and can therefore react with any of a variety of inorganic or organic bases, as well as inorganic and organic acids, to form pharmaceutically acceptable salts. These salts can be prepared by the following method: prepared in situ during the final isolation and purification of the compounds of the present application, or by separately reacting the free base form of the compounds of the present application with a suitable organic or inorganic acid and isolating the salt thus formed. Examples of pharmaceutically acceptable salts include salts prepared by reaction between a compound described herein and an inorganic or organic acid or base. These salts include acetate, acrylate, adipate, alginate, aspartate, benzoate, benzenesulfonate, bisulfate, bisulfite, bromide, butyrate, butyne-1, 4-dicarboxylate, camphorate, camphorsulfonate, caprylate, chlorobenzoate, chloride, citrate, cyclopentanepropionate, caprate, gluconateDihydrogen phosphate, dinitrobenzoate, dodecyl sulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, glycolate, hemisulfate, heptanoate, hexyne-1,6-dioate, hydroxybenzoate, y-hydroxybutyrate, hydrochloride, hydrobromide, hydroiodide, 2-isethionate, iodide, isobutyrate, lactate, maleate, malonate, methanesulfonate, mandelate, metaphosphate, methoxybenzoate, methylbenzoate, monohydrogen phosphate, 1-naphthalenesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, pamoate, jellate (pectate), persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, pyrosulfate, pyrophosphate, propiolic acid, phthalate, phenylacetate, phenylbutyrate, propanesulfonate, salicylate, succinate, sulfate, sulfite, suberate, sebacate, sulfonate, tartrate, thiocyanate, p-toluenesulfonate, ditolute, and ditosylate. Other acids (such as oxalic), while not pharmaceutically acceptable per se, may be used as intermediates in the preparation of the salts to obtain the compounds of the present application and their pharmaceutically acceptable acid addition salts (see, e.g., examples in Berge et al, j. Pharm. Sci.1977,66, 1-19). Further, the compounds described herein, which may include free acid groups, may be reacted with a suitable base (e.g., a hydroxide, carbonate or bicarbonate of a pharmaceutically acceptable metal cation), with ammonia, or with a pharmaceutically acceptable organic primary, secondary or tertiary amine. Representative alkali metal salts or alkaline earth metal salts include lithium, sodium, potassium, calcium, magnesium, and aluminum salts and the like. Illustrative examples of bases include sodium hydroxide, potassium hydroxide, hydroxyethyltrimethylamine hydroxide, sodium carbonate and IV' (C) _1-4 Alkyl radical) ₄ And the like. Representative organic amines useful for the formation of base addition salts include ethylamine, diethylamine, ethylenediamine, ethanolamine, diethanolamine, piperazine and the like. It is to be understood that the compounds described herein also include quats of any basic nitrogen-containing group that they may contain. Water-soluble or oil-soluble compounds obtained by quaternizationOr a dispersible product. See, for example, berge et al, supra.

The term "solvate" refers to a solvate formed by combining a compound of the present application with a solvent molecule. In some embodiments, the solvate is a monohydrate, e.g., the solvent is water, and the compound of the present application forms a monohydrate with water.

Esters of the compounds of the present invention are those pharmaceutically acceptable esters which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of patients without excessive toxicity, irritation, allergic response, and the like. It will be appreciated that esters of compounds of formula I containing a carboxy or hydroxy group include in vivo hydrolysable esters, such as those which are hydrolysed in the human or animal body to form the parent acid or alcohol, which are pharmaceutically acceptable. Suitable pharmaceutically acceptable esters of the carboxyl group formed on the carboxyl group (if present) of the compounds of the invention include, for example, alkyl esters (e.g., C) _1-6 Or C _1-4 Alkyl esters), cycloalkyl esters (e.g. C) _3-12 、C _3-8 Or C _3-6 Cycloalkyl esters), arylalkyl esters (e.g. C) _6-12 Or C _6-8 Arylalkyl esters) and heteroarylalkyl esters (e.g., C) _6-12 Or C _6-8 Heteroarylalkyl esters), and the like.

In vivo hydrolysable esters of compounds of the invention which contain hydroxyl groups include inorganic esters such as phosphate esters, [ alpha ] -acyloxyalkyl ethers and compounds which are related to the result of hydrolytic breakdown of the ester in vivo to give the parent hydroxyl group. Further suitable in vivo hydrolysable esters include those formed from amino acids. Such as esters formed by the reaction of the hydroxyl group of a compound with the carboxylic acid of an amino acid. Further in vivo hydrolysable esters include phosphoramidates, and also include compounds of the present invention wherein any free hydroxyl group independently forms a phosphoryl or phosphite ester.

In addition, the present invention includes all possible crystalline forms or polymorphs of the compounds of the present invention, including individual polymorphs thereof or mixtures of polymorphs in any ratio. The term "polymorph" means that the compounds of the present application exist in different lattice forms.

The term "prodrug" as used herein includes acid derivatives or alcohol derivatives and the like which are well known to practitioners in the art. Among them, acid derivatives such as esters prepared by reaction of the parent acid with a suitable alcohol, or amides prepared by reaction of the parent acid with a substituted or unsubstituted amine, or acid anhydrides, or mixed anhydrides, and the like; the alcohol derivative is selected from, for example, alcohol esters, alcohol alkoxylates, alcohol ethers, carboxylic acids, carboxylic acid esters, phosphonates, mixtures thereof, and the like. Simple aliphatic or aromatic esters, amides and anhydrides resulting from pendant acid groups on the compounds of the invention, as well as phosphates on hydroxyl groups are specific prodrugs.

The term "active metabolite" refers to a biologically active derivative of a compound that is formed when the compound is metabolized.

Certain compounds of the present application may have asymmetric carbon atoms (stereocenters) or double bonds. Thus, racemates, diastereomers, enantiomers, geometric isomers and individual isomers are included within the scope of the present application.

When the compounds of the present application contain olefinic double bonds or other centers of geometric asymmetry, they include both E and Z geometric isomers unless otherwise specified. Likewise, all tautomeric forms are included within the scope of the present application.

The compounds of the present application may exist in specific geometric isomeric or stereoisomeric forms. The present application contemplates all such compounds, including tautomers, cis and trans isomers, (-) -and (+) -enantiomers, (R) -and (S) -enantiomers, diastereomers, (D) -isomers, (L) -isomers, as well as racemic and other mixtures thereof, such as enantiomerically or diastereomerically enriched mixtures, all of which are within the scope of the present application. Additional asymmetric carbon atoms may be present in a substituent such as alkyl. All such isomers and mixtures thereof are included within the scope of the present application.

Optically active (R) -and (S) -isomers as well as D and L isomers can be prepared by chiral synthesis or chiral reagents or other conventional techniques. If one enantiomer of a compound of the present application is desired, it may be prepared by asymmetric synthesis or derivatization with a chiral auxiliary, wherein the resulting diastereomeric mixture is separated and the auxiliary group is cleaved to afford the pure desired enantiomer. Alternatively, where the molecule contains a basic functional group (e.g., amino) or an acidic functional group (e.g., carboxyl), diastereomeric salts are formed with an appropriate optically active acid or base, followed by diastereomeric resolution by conventional methods known in the art, and the pure enantiomers are recovered. Furthermore, separation of enantiomers and diastereomers is typically accomplished by using chromatography employing a chiral stationary phase, optionally in combination with chemical derivatization (e.g., carbamate formation from amines).

The present application also includes isotopically-labeled compounds of the present application, which are identical to those recited herein, but for the fact that one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes that can be incorporated into compounds of the present application include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, iodine and chlorine, such as respectively ² H、 ³ H、 ¹¹ C、 ¹³ C、 ¹⁴ C、 ¹³ N、 ¹⁵ N、 ¹⁵ O、 ¹⁷ O、 ¹⁸ O、 ³¹ P、 ³² P、 ³⁵ S、 ¹⁸ F、 ¹²³ I、 ¹²⁵ I and ³⁶ cl, and the like.

Certain isotopically-labelled compounds of the present application (e.g. with ³ H and ¹⁴ c-labeled ones) can be used in compound and/or substrate tissue distribution assays. Tritiated (i.e. by tritiation) ³ H) And carbon-14 (i.e. ¹⁴ C) Isotopes are particularly preferred for their ease of preparation and detectability. Positron emitting isotopes such as ¹⁵ O、 ¹³ N、 ¹¹ C and ¹⁸ f can be used in Positron Emission Tomography (PET) studies to determine substrate occupancy. Isotopically labeled compounds of the present application can generally be prepared by following procedures analogous to those disclosed in the schemes and/or in the examples below, by substituting an isotopically labeled reagent for a non-isotopically labeled reagent.

In addition, heavier isotopes are used (such as deuterium (i.e., deuterium) ² H) GetSubstitution may provide certain therapeutic advantages resulting from greater metabolic stability (e.g., increased in vivo half-life or reduced dosage requirements), and thus may be preferred in certain circumstances where deuterium substitution may be partial or complete, partial deuterium substitution meaning that at least one hydrogen is substituted with at least one deuterium, all such forms of the compounds being encompassed within the scope of the present application.

The term "subject," "patient," or "individual" refers to an individual suffering from a disease, disorder, or condition, and the like, including mammals and non-mammals. Examples of mammals include, but are not limited to, any member of the class mammalia: humans, non-human primates (e.g., chimpanzees and other apes and monkeys); livestock, such as cattle, horses, sheep, goats, pigs; domestic animals such as rabbits, dogs, and cats; laboratory animals, including rodents, such as rats, mice, and guinea pigs, and the like. Examples of non-human mammals include, but are not limited to, birds, fish, and the like. In one embodiment related to the methods and compositions provided herein, the mammal is a human.

The term "treating" means administering a compound or formulation described herein to ameliorate or eliminate a disease or one or more symptoms associated with the disease, and includes:

(i) Inhibiting the disease or disease state, i.e., arresting its development;

(ii) Alleviating the disease or condition, i.e., causing regression of the disease or condition.

The term "preventing" means administering a compound or formulation described herein to prevent a disease or one or more symptoms associated with the disease, and includes: preventing the occurrence of a disease or condition in an individual, particularly when the individual is predisposed to the disease condition, but has not yet been diagnosed as having the disease condition.

The terms "effective amount," "therapeutically effective amount," or "pharmaceutically effective amount" refer to an amount of at least one agent or compound sufficient to alleviate one or more symptoms of the disease or disorder being treated to some extent upon administration. The result may be a reduction and/or alleviation of signs, symptoms, or causes, or any other desired change in a biological system. For example, an "effective amount" for treatment is the amount of a composition comprising a compound disclosed herein that is clinically necessary to provide a significant remission effect of the condition. An effective amount suitable in any individual case can be determined using techniques such as a dose escalation assay.

The term "acceptable" means that there is no long-term detrimental effect on the general health of the subject being treated.

The term "pharmaceutically acceptable" refers to a substance (e.g., carrier or diluent) that does not affect the biological activity or properties of the compounds of the present application, and is relatively non-toxic, i.e., the substance can be administered to an individual without causing an adverse biological response or interacting in an adverse manner with any of the components contained in the composition.

The term "composition" or "pharmaceutical composition" refers to a biologically active compound optionally mixed with at least one pharmaceutically acceptable chemical ingredient including, but not limited to, carriers, stabilizers, diluents, dispersants, suspending agents, thickeners, and/or excipients.

The term "carrier" refers to a relatively non-toxic chemical compound or agent that facilitates the introduction of the compound into a cell or tissue.

The term "pharmaceutically acceptable adjuvants" refers to those adjuvants which do not have a significant irritating effect on the organism and do not impair the biological activity and properties of the active compound. Suitable adjuvants are well known to those skilled in the art, such as carbohydrates, waxes, water-soluble and/or water-swellable polymers, hydrophilic or hydrophobic materials, gelatin, oils, solvents, water, and the like.

The word "comprise" and its derivatives, such as "comprises" or "comprising," are to be construed in an open, non-exclusive sense, i.e., to mean "including, but not limited to.

The pharmaceutical compositions of the present application can be prepared by combining the compounds of the present application with suitable pharmaceutically acceptable excipients, for example, can be formulated into solid, semi-solid, liquid or gaseous formulations such as tablets, pills, capsules, powders, granules, lozenges, ointments, syrups, emulsions, suspensions, solutions, suppositories, injections, inhalants, gels, microspheres, aerosols, and the like.

Typical routes of administration or administration of the compounds of the present application, pharmaceutically acceptable salts, esters, solvates, prodrugs, active metabolites, crystals, stereoisomers, tautomers or geometric isomers thereof, or pharmaceutical compositions thereof, include, but are not limited to, oral, rectal, transmucosal, topical, transdermal, inhalation, parenteral, sublingual, intravaginal, intranasal, intraocular, intraperitoneal, intramuscular, subcutaneous, and intravenous administration. Preferred routes of administration are oral administration and injection.

The compounds or compositions of the present invention may be formulated and used as the following dosage forms: tablets, capsules or elixirs for oral administration; suppositories for rectal administration; sterile solutions for administration by injection, suspensions; patches for transdermal administration, subcutaneous deposits, and the like. Injections can be prepared in the following conventional forms: solutions or suspensions, solid dosage forms suitable for constitution with a solution or suspension prior to injection, or emulsions. Suitable excipients are, for example, water, saline, dextrose, mannitol, lactose, lecithin, albumin, sodium glutamate, cysteine hydrochloride and the like. In addition, if desired, the injectable pharmaceutical compositions may contain minor amounts of non-toxic adjuvants such as wetting agents, pH buffering agents and the like. Absorption enhancers (e.g., liposomes) can also be used if desired.

Formulations for parenteral administration include aqueous solutions of the active compounds in water-soluble form. Alternatively, suspensions of the active compounds may be prepared as appropriate oily injection suspensions. Suitable lipophilic solvents or vehicles include fatty oils such as sesame oil, or other organic oils such as soybean oil, grapefruit oil or almond oil, or synthetic fatty acid esters such as ethyl oleate or triglycerides, or liposomes. Aqueous injection suspensions may contain substances which increase the viscosity of the suspension, for example sodium carboxymethyl cellulose, sorbitol, or dextran. Optionally, the suspension may contain suitable stabilizers or agents that increase the solubility of the compound, allowing for the preparation of highly concentrated solutions.

The pharmaceutical compositions of the present application can be prepared by methods known in the art, such as conventional mixing, dissolving, granulating, dragee-making, levigating, emulsifying, and lyophilizing processes.

In some embodiments, the pharmaceutical compositions of the present application are in oral form. For oral administration, the pharmaceutical compositions may be formulated by mixing the active compound with pharmaceutically acceptable adjuvants or excipients well known in the art. These adjuvants or excipients enable the compounds of the present application to be formulated as tablets, pills, dragees, capsules, powders, granules, liquids, syrups, emulsions, gels, slurries, suspensions and the like, for oral administration to a patient.

Solid pharmaceutical compositions suitable for oral administration may be prepared by conventional mixing, filling or tableting methods. For example, oral compositions in solid form can be obtained by: the active compound is mixed with solid adjuvants or excipients, the mixture obtained is optionally milled, if desired with further suitable adjuvants or excipients, and the mixture is then processed to granules, to give tablets or dragee cores. Suitable adjuvants or excipients include, but are not limited to: fillers, binders, diluents, disintegrants, lubricants, glidants, sweeteners or flavoring agents and the like. Pharmaceutical preparations for oral use, for example, can be obtained by the following process: combining the active compound with a solid excipient, optionally grinding the resulting mixture, and processing the mixture of granules, if desired after addition of suitable auxiliaries, to give tablets or dragee cores. Suitable excipients are, in particular, fillers such as sugars, including lactose, sucrose, mannitol or sorbitol; cellulose preparations such as corn starch, wheat starch, rice starch, potato starch, gelatin, tragacanth, methyl cellulose, hydroxypropylmethyl-cellulose, sodium carboxymethylcellulose, and/or polyvinylpyrrolidone (PVP). If desired, disintegrating agents may be added, for example cross-linked polyvinylpyrrolidone, agar or alginic acid or alginates such as sodium alginate. The dragee cores are suitably coated. For this purpose, concentrated sugar solutions can be used, which can optionally contain gum arabic, talc, polyvinylpyrrolidone, carbopol gel, polyethylene glycol, and/or titanium dioxide, lacquer solutions, and suitable organic solvents or solvent mixtures. Dyes or pigments may be added to the tablets or dragee coatings in order to identify or characterize different combinations of active compound doses. These formulations can be made using methods well known in the art.

In all methods of administration of the compounds or compositions described herein, the daily dose administered may be, for example, from 0.001 to 300mg/kg body weight, such as from 0.01 to 300mg/kg body weight, or from 10 to 200mg/kg body weight, administered as a single dose or in divided doses.

The pharmaceutical compositions of the present application may also be adapted for parenteral administration, such as sterile solutions, suspensions, emulsions or lyophilized products in suitable unit dosage forms. Suitable excipients, such as fillers, buffers or surfactants can be used.

The application takes Bcr-Abl and BTK as targets, develops a kinase inhibitor and is used for treating diseases such as tumor nervous system diseases, rheumatoid arthritis and the like.

wherein:

R ₀ selected from the group consisting of halogen (e.g., fluorine, chlorine, bromine, iodine) and D;

R ₁ 、R ₄ and R ₆ Each independently selected fromOptionally substituted alkyl, optionally substituted hydroxyalkyl, halogen, optionally substituted alkoxy, optionally substituted alkynyl, optionally substituted alkenyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted carboxyalkyl, -NO ₂ -CN, -OH, -COOH, thiol, optionally substituted amino, optionally substituted alkylthio, optionally substituted ester, optionally substituted acyl, and optionally substituted sulfonyl;

In some embodiments, R ₁ 、R ₄ And R ₆ Each independently selected from optionally substituted C _1-6 Alkyl, optionally substituted C _1-6 Hydroxyalkyl, halogen, optionally substituted C _1-6 Alkoxy, optionally substituted C _2-6 Alkynyl, optionally substituted C _2-6 Alkenyl, optionally substituted 3-8 membered (e.g., 3, 4, 5, 6, 7, 8 membered) cycloalkyl, optionally substituted 3-10 membered (e.g., 3, 4, 5, 6, 7, 8, 9, 10 membered) heterocyclyl, optionally substituted 6-10 (e.g., 6, 7, 8, 9, 10 membered) aryl, optionally substituted 5-10 membered (e.g., 5, 6, 7, 8, 9, 10 membered) heteroaryl, optionally substituted C _1-6 Carboxyalkyl, -NO ₂ -CN, -OH, -COOH, mercapto, optionally substituted amino, optionally substituted C _1-6 Alkylthio, optionally substituted ester, optionally substituted acyl and optionally substituted sulfonyl.

In some embodiments, R ₈ Selected from H, D, optionally substituted C _1-6 Hydroxyalkyl, optionally substituted C _1-6 Carboxyalkyl, optionally substituted C _1-6 Alkyl, halogen, optionally substituted C _1-6 Alkoxy radicalOptionally substituted C _2-6 Alkynyl, optionally substituted C _2-6 Alkenyl, optionally substituted 3-8 membered (e.g., 3, 4, 5, 6, 7, 8 membered) cycloalkyl, optionally substituted 3-10 membered (e.g., 3, 4, 5, 6, 7, 8, 9, 10 membered) heterocyclyl, optionally substituted 6-10 membered (e.g., 6, 7, 8, 9, 10 membered) aryl, optionally substituted 5-10 membered (e.g., 5, 6, 7, 8, 9, 10 membered) heteroaryl, -NO ₂ -CN, -OH, -COOH, mercapto, optionally substituted amino, optionally substituted C _1-6 Alkylthio, optionally substituted ester, optionally substituted acyl and optionally substituted sulfonyl.

In some embodiments, R ₁ 、R ₄ And R ₆ Each independently selected from optionally substituted C _1-6 Alkyl (e.g. C) ₁ 、C ₂ 、C ₃ 、C ₄ 、C ₅ 、C ₆ ) Optionally substituted C _1-6 Hydroxyalkyl, and halogen.

In some embodiments, R ₁ Is selected from C _1-4 Alkyl radical, C _1-4 Haloalkyl, C _1-4 Hydroxyalkyl and halogen (e.g., fluorine, chlorine, bromine, iodine).

In some embodiments, R ₁ Is selected from-CH ₃ 、-CH ₂ OH and chlorine.

In some casesIn embodiments, R ₁ Is selected from-CH ₃ And chlorine.

In some embodiments, R ₄ Is selected from C _1-4 Alkyl radical, C _1-4 Haloalkyl, C _1-4 Deuterated alkyl and halogen (e.g., fluorine, chlorine, bromine, iodine).

In some embodiments, R ₄ Is selected from-CH ₃ Fluorine and chlorine.

In some embodiments, R ₄ Is selected from-CH ₃ And chlorine.

In some embodiments, R ₆ Is selected from C _1-4 An alkyl group.

In some embodiments, R ₆ Is selected from-CH ₃ 、-CH ₂ OH and-CD ₃ 。

In some embodiments, R ₆ Is selected from-CH ₃ 。

In some embodiments, R ₈ Is selected from C _1-4 A hydroxyalkyl group.

In some embodiments, R ₈ Is selected from-CH ₂ CH ₂ OH。

in yet another aspect, the present application provides a pharmaceutical composition comprising at least any one of the compounds provided in the present disclosure, or a pharmaceutically acceptable salt, ester, solvate, prodrug, active metabolite, crystal, stereoisomer, tautomer, or geometric isomer thereof. In some embodiments, the compositions provided herein further comprise a pharmaceutically acceptable carrier, excipient, or adjuvant. In some embodiments, the pharmaceutical composition of the present disclosure further comprises another therapeutic agent for combination therapy. In some embodiments, the additional therapeutic agent is selected from one or more of the following: cyclophosphamide, ifosfamide, vincristine, daunorubicin, doxorubicin, cytarabine, mitoxantrone, dacarbazine, idarubicin, retinoic acid, prednisone, dexamethasone, mercaptopurine, methotrexate, paclitaxel, melphalan, long-acting interferon, venetocel, crizotinib, erlotinib, ocitinib, ruxotinib, afatinib, erlotinib, imatinib, lapatinib, bevacizumab, trastuzumab, rituximab, cetuximab, bornatuzumab, fludarabine, gemcitabine, decitabine, capecitabine, bendamustine, everolimus, temsirolimus, etoposide, granulocyte colony stimulating factor, temozolomide, zoledronic acid, olixaplatin, cisplatin, carboplatin, and fulvestrant.

In another aspect, the present application provides a compound of the present disclosure, or a pharmaceutically acceptable salt, ester, solvate, prodrug, active metabolite, crystal, stereoisomer, tautomer, or geometric isomer thereof, or a pharmaceutical composition thereof, for use in preventing or treating a tyrosine kinase-associated disease.

In yet another aspect, the present application provides the use of a compound described in the present disclosure, or a pharmaceutically acceptable salt, ester, solvate, prodrug, active metabolite, crystal, stereoisomer, tautomer, or geometric isomer thereof, or a pharmaceutical composition thereof, for the manufacture of a medicament for inhibiting tyrosine kinase activity.

In yet another aspect, the present application provides a method of inhibiting tyrosine kinase activity, comprising administering to a subject or a tissue or cell thereof at least any one of the compounds provided by the present disclosure or a pharmaceutically acceptable salt, ester, solvate, prodrug, active metabolite, crystal, stereoisomer, tautomer, or geometric isomer thereof, or a pharmaceutical composition thereof. In some embodiments, the method of inhibiting tyrosine kinase activity comprises an in vitro or in vivo assay.

In another aspect, the present disclosure provides a compound described herein, or a pharmaceutically acceptable salt, ester, solvate, prodrug, active metabolite, crystal, stereoisomer, tautomer, or geometric isomer thereof, or a pharmaceutical composition thereof, for use in inhibiting tyrosine kinase activity.

In another aspect, the present disclosure provides a compound described in the present disclosure, or a pharmaceutically acceptable salt, ester, solvate, prodrug, active metabolite, crystal, stereoisomer, tautomer, or geometric isomer thereof, or a pharmaceutical composition thereof, for use in the prevention or treatment of a tyrosine kinase associated disease or disorder.

In some embodiments, the disease is selected from cancer.

In some embodiments, the cancer is selected from: chronic Myelogenous Leukemia (CML), gastrointestinal stromal tumor (GIST), small Cell Lung Cancer (SCLC), non-small cell lung cancer (NSCLC), multiple myeloma, solid tumors, B-cell lymphoma, chronic Lymphocytic Leukemia (CLL), acute Lymphocytic Leukemia (ALL), non-Hodgkin's lymphoma (NHL), small Lymphocytic Lymphoma (SLL), mantle Cell Lymphoma (MCL), melanoma, mastocytosis, germ cell tumors, acute Myeloid Leukemia (AML), marginal zone/diffuse large B-cell lymphoma, sarcoma, pancreatic cancer, glioblastoma, head and neck tumors, macroglobulinemia, follicular central lymphoma, prostate cancer, myelodysplastic syndrome, atherosclerotic myelohyperplasia, myelofibrosis, eosinophilia, polycythemia vera, liver cancer, advanced sarcoma, glioblastoma multiforme, gliosarcoma, malignant mesothelioma, melanoma, squamous cell carcinoma skin cancer, neuroendocrine tumor, gastric tumor, B-cell acute lymphocytic leukemia, hairy cell leukemia, lymphoplasmacytic lymphoma, follicular central lymphoma, renal cell carcinoma, transitional cell carcinoma, carcinoid tumor, T-cell lymphoma, metastatic non-small cell lung cancer, systemic mastocytosis, metastatic renal cell carcinoma, breast tumor, central nervous system tumor, colorectal tumor, metastatic bladder cancer, metastatic pancreatic cancer, metastatic head and neck cancer, ovarian tumor and combinations thereof.

In some embodiments, the cancer is selected from the group consisting of cancer that is resistant to the chemotherapeutic agent that targets BCR-ABL and c-KIT, and cancer that is resistant to imatinib.

In some embodiments, the diseases, disorders, and conditions are selected from: bone metastasis, hypercalcemia and/or osteoporosis; pulmonary fiber diseases; cardiovascular disease or condition; mast cell mediated inflammatory diseases; HTLV-1 associated myelopathy/tropical spastic paralysis; complex Regional Pain Syndrome (CRPS); weight loss or fat loss; arterial occlusive disease; ubiquitination; a disease or condition associated with reduced carbohydrate degrading function; fridriich ataxia; graft rejection in parkinson's disease progression; rheumatoid arthritis; graft versus host disease; (ii) an autoimmune disease; recurrent immune thrombocytopenic purpura, pemphigus vulgaris, systemic lupus erythematosus, scleroderma pulmonary interstitial fibrosis and idiopathic urticaria.

In some embodiments, the cardiovascular disease or condition is a cardiovascular disease caused by RASopathy, or a congenital heart disease associated with nosan or nosan syndrome.

The inventions of the present application provide one or more of the following advantages:

1. the compounds of the present disclosure have kinase inhibitory activity comparable to or better than existing drugs such as dasatinib.

2. In vitro liver microsome assay showed: compared with the existing medicines such as dasatinib, the compound of the formula I disclosed by the invention has longer half-life (> 60 h) and lower clearance rate. While the current dose of dasatinib is 100mg daily, 1-2 times daily, the compounds of formula I of the present disclosure may be administered at lower doses and at longer dosing intervals than the currently used drugs, such as dasatinib.

At nanomolar concentrations (nM), the inhibitor compounds of the present disclosure inhibit the SRC kinase family ABL1, BTK and BTK (C481S) kinases, and by inhibiting the actions of these kinases, the proliferation of leukemic cells in the bone marrow of CML and Ph + ALL can be inhibited, but normal erythrocytes, leukocytes and platelets can continue to proliferate.

The invention improves the curative effect of the medicine, reduces the dosage, thereby reducing the toxic and side effect and better providing clinical treatment for diseases such as tumor, rheumatoid arthritis and the like. The novel low-dose inhibitor disclosed by the invention can provide a wide application prospect for treating diseases.

Detailed Description

The following examples are given to further illustrate embodiments of the present invention. The following examples are for illustrative purposes only and are not intended to limit the scope of the present application.

Preparation example 1

Synthesis of Compound 1:

the synthetic route for compound 1 is as follows:

synthesis of [5- (2-chloro-4-fluoro-6-methylphenylcarbamoyl) thiazol-2-yl ] -carbamic acid tert-butyl ester

N ₂ 24.4g (0.1 mol) of 2-tert-butoxycarbonylaminothiazole-5-carboxylic acid and 0.5ml of DMF (N, N-dimethylformamide) were added to 250ml of dichloromethane under a gas blanket, 13ml of oxalyl chloride solution (0.15 mol) were slowly added dropwise, the reaction was carried out for 2 hours, the solvent was removed by rotary evaporation to give a white solid, the solid was dissolved in 100ml of anhydrous dichloromethane, and the solution was slowly added dropwise to a solution of 17.5g (0.11 mol) of 2-chloro-4-fluoro-6-methylaniline and 38.8g (0.3 mol) of N, N-diisopropylethylamine in dichloromethane under an ice bath condition ₂ Reacting at room temperature for 10H under the protection of gas, distilling under reduced pressure to remove the solvent, adding a mixed solvent of 25ml ethyl acetate and 25ml n-hexane, stirring for 2H, filtering, leaching with ethyl acetate to obtain off-white powdery solid 35.2g, yield 85%, purity 95% (ESI-MS (M/z): [ M + H ])] ⁺ ，386。 ¹ H NMR(DMSO-d ₆ )，δ：1.49(s，9H，-CH ₃ )，2.18(s，3H，-CH ₃ ) 6.72 (s, 1H, aromatic hydrogen), 6.78 (s, 1H, aromatic hydrogen), 8.14 (s, 1H, thiazolehydrogen), 9.51 (s, 1H, -NH), 11.81 (s, 1H, -NH).

Synthesis of 2-amino-N- (2-chloro-4-fluoro-6-methylphenyl) -5-thiazolecarboxamide

Reacting [5- (2-chloro-4-fluoro-6-methylphenylcarbamoyl) thiazol-2-yl]Adding 33.6g (0.087 mol) of tert-butyl carbamate into 200ml of dichloromethane, adding 100ml of trifluoroacetic acid, stirring for 6h at room temperature, detecting by TLC for complete reaction, concentrating under reduced pressure, adding the obtained oily substance into ice water, adjusting the pH value to 10 by using NaOH solution, fully stirring until no oily substance exists, adjusting the pH value to 7 again, separating out a large amount of solid, performing suction filtration, leaching and drying to obtain 23.0g of light yellow solid, wherein the yield is 92.5%. ESI-MS (m/z): [ M + H ]] ⁺ ，286。 ¹ HNMR(DMSO-d ₆ )，δ：2.21(s，3H，-CH ₃ ) 6.71 (s, 1H, aromatic Hydrogen), 6.76 (s, 1H, aromatic Hydrogen), 7.63 (s, 2H, -NH ₂ ) 7.88 (s, 1H, thiazolehydrogen), 9.66 (s, 1H, -NH).

Synthesis of N- (2-chloro-4-fluoro-6-methylphenyl) -2- [ (2-methyl-6-chloro-4-pyrimidinyl) amino ] -5-thiazolecarboxamide

Dissolving 21.3g (0.075 mol) of 2-amino-N- (2-chloro-4-fluoro-6-methylphenyl) -5-thiazolecarboxamide in 100ml of DMF, adding 17.93g (0.11 mol) of 2-methyl-4, 6-dichloropyrimidine and 48.9g (0.15 mol) of cesium carbonate, carrying out heat preservation reaction at 40 ℃ for 12 hours, carrying out suction filtration, adding the filtrate into 300ml of ice water, adjusting the pH value to 6 by using diluted hydrochloric acid, carrying out stirring crystallization, carrying out suction filtration, carrying out rinsing on the filter cake by using ethyl acetate, and drying to obtain 29.5g of light yellow solid, wherein the yield is 95.3%. ESI-MS (m/z): [ M + H ]] ⁺ ，412。 ¹ H NMR(DMSO-d ₆ )，δ：2.25(s，3H，-CH ₃ )，2.59(s，3H，-CH ₃ ) 6.98 (s, 1H, pyrimidinehydrogen), 6.73 (s, 1H, aromatic hydrogen), 6.77 (s, 1H, aromatic hydrogen), 8.32 (s, 1H, thiazolehydrogen), 10.01 (s, 1H, -NH), 12.21 (s, 1H, -NH).

Synthesis of N- (2-chloro-4-fluoro-6-methylphenyl) -2- [ [6- [4- (2-hydroxyethyl) -1-piperazinyl ] -2-methyl-4-pyrimidinyl ] amino ] -5-thiazolecarboxamide

Weighing N- (2-chloro-4-fluoro-6-methylphenyl) -2- [ (2-methyl-6-chloro-4-pyrimidinyl) amino]28.2g (0.068 mol) of-5-thiazolecarboxamide, 44.3g (0.34 mol) of N-hydroxyethylpiperazine and 26.4g (0.204 mol) of N, N-diisopropylethylamine were dissolved in 250ml of isopropanol, the mixture was heated to 83 ℃ and refluxed for 8 hours, a part of the solvent was concentrated under reduced pressure, cooled to room temperature, filtered, and the filter cake was recrystallized from a mixed solution of ethanol and water 1 to obtain 31.36g of a white powdery solid, the yield was 91.1%, and the purity was 98%. ESI-MS (m/z): [ M + H ]] ⁺ ，506。 ¹ HNMR(DMSO-d ₆ )，δ：2.21(s，3H，-CH ₃ )，2.41(s，3H，-CH ₃ )，2.43(t，2H，-CH ₂ )，2.48(t，4H，-CH ₂ )，3.52-3.54(m，4H，-CH ₂ )，3.55-3.56(m，2H，-CH ₂ ) 4.48 (s, 1H, -OH), 6.06 (s, 1H, pyrimidinehydrogen), 6.73 (s, 1H, aromatic hydrogen), 6.77 (s, 1H, aromatic hydrogen), 8.22 (s, 1H, thiazolinehydrogen), 9.87 (s, 1H, -NH), 11.45 (s, 1H, -NH).

Synthesis of N- (2-chloro-4-fluoro-6-methylphenyl) -2- [ [6- [ 4-oxo-4- (2-hydroxyethyl) -1-piperazinyl ] -2-methyl-4-pyrimidinyl ] amino ] -5-thiazolecarboxamide

Weighing N- (2-chloro-4-fluoro-6-methylphenyl) -2- [ [6- [4- (2-hydroxyethyl) -1-piperazinyl]-2-methyl-4-pyrimidinyl]Amino group]30.4g (0.06 mol) of-5-thiazolecarboxamide is dissolved in 200mL of dichloromethane, 8.2g (0.072 mol) of 30% hydrogen peroxide is dropwise added at normal temperature, the reaction is continued for 24 hours after the dropwise addition is finished, the solution is subjected to layering, water washing, organic layer decompression and concentration, and the residue is recrystallized by 85% ethanol to obtain 22.3g of white powdery solid, the yield is 71.0%, and the purity is 98%. ESI-MS (m/z): [ M + H ]] ⁺ ，522。 ¹ H NMR(DMSO-d ₆ ) δ:2.16 (m, 3H, phenyl ring-CH) ₃ ) 2.41 (m, 3H, pyrimidine ring-CH) ₃ )，3.30(m，8H，CH ₂ )，3.61(m，2H，CH ₂ )，3.90(s，2H，CH ₂ ) 4.08 (m, 2H, active hydrogen), 6.10 (s, 1H, pyrimidinehydrogen), 7.21 (d, J =6.8,1h, aromatic hydrogen), 7.38 (d, J =5.6,1h, aromatic hydrogen), 8.20 (s, 1H, thiazolehydrogen), 9.83 (s, 1H, active hydrogen).

Preparation example 2

Synthesis of Compound 2:

the synthetic route for compound 2 is as follows:

synthesis of 2- (4- (6-chloro-2-methylpyrimidin-4-yl) piperazin-1-yl) ethanol

2- (piperazin-1-yl) ethanol (13.0 g, 100mmol) and triethylamine (30.3 g, 300mmol) were added to a solution of 4, 6-dichloro-2-dimethylpyrimidine (16.3 g, 100mmol) in 400mL of dichloromethane and stirred at room temperature for 2 hours (reaction monitored by LCMS). After completion of the reaction, the reaction mixture was washed with saturated brine (50 ml. Times.1) and water (50 ml. Times.1), respectively, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated in vacuo to give 30.8g of a yellow powdery solid, yield 82%, purity 68%. ESI-MS (m/z): [ M + H ]] ⁺ ，257。

Synthesis of methyl 2- ((6- (4- (2-hydroxyethyl) piperazin-1-yl) -2-methylpyrimidin-4-yl) amino) thiazole-5-carboxylate

The compound 2- (4- (6-chloro-2-methylpyrimidin-4-yl) piperazin-1-yl) ethanol (30.8 g crude, 100 mol), SM-2 (20.5g, 130mmol), pd ₂ (dba) ₃ (0.916g，1mmol)，Xantphos(1.418g，2mmol)，And Cs ₂ CO ₃ (97.8g, 300mmol) was added to toluene (400 ml) and the mixture was heated to 120 ℃ under argon and stirred for 4 hours. After the reaction is completed, the reaction solution is concentrated in vacuum to obtain a solid mixture, water and petroleum ether are sequentially used for pulping, and the yellow powdery solid of 40g is obtained by drying, wherein the yield is 100%, and the purity is 77%. ESI-MS (m/z): [ M + H ]] ⁺ ，379。

Synthesis of 2- ((6- (4- (2-hydroxyethyl) piperazin-1-yl) -2-methylpyrimidin-4-yl) amino) thiazole-5-carboxylic acid

LiOH (9.6 g, 400mol) was added to a methanol/water mixture (200/200 mL) of the compound methyl 2- ((6- (4- (2-hydroxyethyl) piperazin-1-yl) -2-methylpyrimidin-4-yl) amino) thiazole-5-carboxylate (40 g crude, 100 mol), and stirred at 60 ℃ for 4 hours. After cooling, 1M hydrochloric acid was added to adjust pH =3. The methanol was then removed by vacuum concentration and rotary evaporation. Water (10 ml) was then added and the solid was filtered and dried to give 33g of a yellow powder solid in 100% yield and 90% purity. ESI-MS (m/z): [ M + H ]] ⁺ ，365。

Synthesis of 2- ((6- (4- (2-acetoxyethyl) piperazin-1-yl) -2-methylpyrimidin-4-yl) amino) thiazole-5-carboxylic acid

Triethylamine (2.70g, 27.0 mmol) and acetic anhydride (1.84g, 18.0 mmol) were added to a solution of the compound methyl 2- ((6- (4- (2-hydroxyethyl) piperazin-1-yl) -2-methylpyrimidin-4-yl) amino) thiazole-5-carboxylate (3.3 g crude, 9.00 mmol) in dichloromethane (36 mL) and stirred at room temperature for 2 hours. Water (10 mL) was then added and the mixture was extracted with dichloromethane (10 mL). Then dried over anhydrous sodium sulfate, filtered and concentrated to give a solid mixture, which was spin-dried to give 3.65g of a yellow powdery solid in 90% yield and 80% purity. ESI-MS (m/z): [ M + H ]] ⁺ ，407。

Preparation of the compound 2- (4- (6- ((5- ((4-fluoro-2, 6-dimethylphenyl) carbamoyl) thiazol-2-yl) amino) -2-methylpyrimidin-4-yl) piperazin-1-yl) ethyl acetate and the compound N- (4-fluoro-2, 6-dimethylphenyl) -2- ((6- (4- (2-hydroxyethyl) piperazin-1-yl) -2-methylpyrimidin-4-yl) amino) thiazole-5-carboxamide

Triethylamine (303mg, 3.00mmol) and HATU (570mg, 1.50mmol) were added to a solution of compound 2- ((6- (4- (2-acetoxyethyl) piperazin-1-yl) -2-methylpyrimidin-4-yl) amino) thiazole-5-carboxylic acid (406 mg, 1.00mmol) in DMF (4 mL), and after stirring at room temperature for 30 minutes, 4-fluoro-2, 6-dimethylaniline (209mg, 1.50mmol) was added and further stirring was carried out for 30 minutes. After completion of the amide condensation reaction (reaction monitored by LCMS), a saturated aqueous solution of LiOH (2 ml) was added, the mixture was stirred at room temperature overnight, and after completion of the reaction (reaction monitored by LCMS), a saturated aqueous solution of ammonium chloride (10 ml) was added to precipitate a solid, which was then filtered, washed with water, and dried to obtain 300mg of a yellow powdery solid, with a yield of 62% in two steps and a purity of 97%. ESI-MS (m/z): [ M + H ]] ⁺ ，486。

Will H ₂ O ₂ (30% strength, 2 ml) was added to a solution of the compound N- (4-fluoro-2, 6-dimethylphenyl) -2- ((6- (4- (2-hydroxyethyl) piperazin-1-yl) -2-methylpyrimidin-4-yl) amino) thiazole-5-carboxamide (300mg, 0.618mmol) in NMP (2 ml), stirred at room temperature for 24 hours, purified by high performance liquid chromatography (elution system 0.1% ammonia/acetonitrile), concentrated and lyophilized to give compound 2 in the free form of a white solid, 80mg, yield 26%, purity 97%. ESI-MS (m/z): [ M + H ]] ⁺ ，502。 ¹ H NMR(400MHz，DMSO-d ₆ ) δ:2.19 (s, 6H, phenyl ring-CH) ₃ ) 2.44 (s, 3H, pyrimidine ring-CH) ₃ )，3.28-3.39(m，8H，CH ₂ )，3.65(t，J＝10.7，2H，CH ₂ )，3.94(s，2H，CH ₂ ) 4.12 (br, 2H, active hydrogen), 6.16 (s, 1H, pyrimidinehydrogen), 6.99 (d, J =9.6,2h, aromaticHydrogen), 8.21 (s, 1H, thiazolehydrogen), 9.64 (s, 1H, active hydrogen).

Preparation example 3

Synthesis of Compound 3:

the synthetic route for compound 3 is as follows:

2- ((6- (4- (2-acetoxyethyl) piperazin-1-yl) -2-methylpyrimidin-4-yl) amino) thiazole-5-carboxylic acid was prepared according to Synthesis procedures 1-4 of Compound 2.

Synthesis of compound 4-fluoro-2-isopropenylaniline

Under the protection of argon, 2-bromo-4-fluoroaniline (10.0g, 52.6mmol), isopropenylboronic acid pinacol ester (9.7g, 57.8mmol) and K are added ₂ CO ₃ (21.7g, 157.8mmol) and Pd ₂ (dppf) ₂ Cl (3.8g, 5.2mmol) was added to 250ml of a 1, 4-dioxane/water (10. After completion of the reaction, water (100 ml) was added and extracted with ethyl acetate (100 ml. Times.3). The organic phases were combined, dried and concentrated to give the crude product. Chromatography column with ethyl acetate/petroleum ether (50): 56.6 percent. ESI-MS (m/z): [ M + H ]] ⁺ ，152。

Synthesis of compound 4-fluoro-2-isopropyl aniline

The compound 4-fluoro-2-isopropenylaniline (4.5g, 29.7 mmol) was added to 100ml of methanol, the air in the reaction flask was replaced with hydrogen, and 500mg of 10% palladium on carbon was added thereto, and the reaction was carried out overnight at room temperature and examined by LCMS. After the reaction is finished, palladium-carbon is filtered out, and the filtrate is concentrated to obtain 4.0g of blue liquid productThe yield is as follows: 87.7 percent. ESI-MS (m/z): [ M + H ]] ⁺ ，154。

Synthesis of compound 2-chloro-4-fluoro-6-isopropylaniline

Compound 4-fluoro-2-isopropylaniline (4 g, 26.1mmol) was dissolved in 100ml of acetonitrile, NCS (4.1g, 31.3mmol) was added, and the reaction was heated to 85 ℃ for 1 hour and examined by LCMS. After the reaction is finished, 100ml of water is added and the mixture is stirred for 10min, then extracted by ethyl acetate (100 ml × 3), the organic phases are combined, dried and concentrated to obtain a crude product, and the crude product is filtered by a chromatographic column by adopting ethyl acetate/petroleum ether (50): 53.2 percent. ESI-MS (m/z): [ M + H ]] ⁺ ，188。

Synthesis of the compound ethyl 2- (4- (6- ((5- ((2-chloro-4-fluoro-6-isopropylphenyl) carbamoyl) thiazol-2-yl) amino) -2-methylpyrimidin-4-yl) piperazin-1-yl) acetate

The compound 2- ((6- (4- (2-acetoxyethyl) piperazin-1-yl) -2-methylpyrimidin-4-yl) amino) thiazole-5-carboxylic acid (300mg, 0.73mmol) and pyridine (50 ml) were charged to a 100ml single-necked flask, the atmosphere in the flask was replaced with argon gas, the reaction flask was placed in an ice-water bath to cool (10 min), and 0.9ml of POCl was added to the reaction flask ₃ After stirring for 10min, the compound 4-fluoro-2-isopropylaniline (205mg, 1.09mmol) was added to the reaction flask. After 1h of reaction, detection by LCMS showed complete reaction, addition of 50ml water and stirring, extraction with ethyl acetate (50 ml × 3), combination of the organic phases, washing with saturated brine (50 ml × 2), drying of the organic phase and concentration to give crude product, which was dissolved with methanol (15 ml), and chromatography to give off-white solid product 150mg, yield: 35.2 percent and the purity is 92 percent. ESI-MS (m/z): [ M + H ]] ⁺ ，576。

Synthesis of compound N- (2-chloro-4-fluoro-6-isopropylphenyl) -2- ((6- (4- (2-hydroxyethyl) piperazin-1-yl) -2-methylpyrimidin-4-yl) amino) thiazole-5-carboxamide

Ethyl 2- (4- (6- ((5- ((2-chloro-4-fluoro-6-isopropylphenyl) carbamoyl) thiazol-2-yl) amino) -2-methylpyrimidin-4-yl) piperazin-1-yl) acetate (150mg, 0.26mmol) was dissolved in 10ml of methanol, the reaction flask was placed in an ice-water bath, a saturated LiOH solution (LiOH: 100 mg) was added, and after 0.5h of reaction, the reaction was completed by LCMS detection, concentrated and dried to give a crude product of 200mg with a purity of 85%. ESI-MS (m/z): [ M + H ]] ⁺ ，534。

Synthesis of the compound 4- (6- ((5- ((2-chloro-4-fluoro-6-isopropylphenyl) carbamoyl) thiazol-2-yl) amino) -2-methylpyrimidin-4-yl) -1- (2-hydroxyethyl) piperazine 1-oxide

H is to be ₂ O ₂ (30% strength, 2 ml) was added to a solution of the compound N- (2-chloro-4-fluoro-6-isopropylphenyl) -2- ((6- (4- (2-hydroxyethyl) piperazin-1-yl) -2-methylpyrimidin-4-yl) amino) thiazole-5-carboxamide (200 mg, crude) in NMP (10 ml), stirred at room temperature for 24 hours, purified by high performance liquid chromatography (eluent water/acetonitrile), concentrated and lyophilized to give 60mg of a white solid, yield 41.9%, purity 98.7%. ESI-MS (m/z): [ M + H ]] ⁺ ，550。 ¹ H NMR(400MHz，DMSO-d ₆ )，δ：1.15(d，J＝6.8Hz，6H，CH ₃ )，2.42(s，3H，-CH ₃ )，3.25(m，1H，-CH)，3.27(m，8H，-CH ₂ )，3.65(t，J＝22.4Hz，2H，-CH ₂ )，3.92(s，2H，-CH ₂ ) 4.11 (d, J =11.2hz,2h, active hydrogen), 6.09 (d, 1H, pyrimidinehydrogen), 7.25-7.22 (m, 1H, aromatic hydrogen), 7.41-7.38 (m, 1H, aromatic hydrogen), 8.20 (s, 1H, thiazolehydrogen), 9.77 (s, 1H, -OH).

Preparation example 4

Synthesis of Compound 4:

the synthetic route for compound 4 is as follows:

Synthesis of the compound 2- (4- (6- ((5- ((2-chloro-4, 6-difluorophenyl) carbamoyl) thiazol-2-yl) amino) -2-methylpyrimidin-4-yl) piperazin-1-yl) ethyl acetate

The compound 2- ((6- (4- (2-acetoxyethyl) piperazin-1-yl) -2-methylpyrimidin-4-yl) amino) thiazole-5-carboxylic acid (550mg, 1.354mmol) and the compound 2-chloro-4, 6-difluoroaniline (243mg, 1.489mmol) were added to a solution of pyridine (5 mL) and POCl was added dropwise at 0 ℃ in an ice bath ₃ (0.5 mL), stir at room temperature for 60 minutes, after the reaction was complete (reaction monitored by LCMS), add ethyl acetate (15 mL) and water (10 mL), saturated NaHCO ₃ Adjusting pH of the aqueous solution to 6-7, extracting the aqueous phase with ethyl acetate (15 mL × 3), washing the organic phase with water (25 mL × 3) and saturated brine (25 mL × 1), drying with anhydrous sodium sulfate, filtering, concentrating to obtain brown oily crude product, purifying with high performance liquid chromatography (mobile phase acetonitrile/water), concentrating, and lyophilizing to obtain white solid 150mg, with yield: 20.1 percent and the purity is 97.5 percent. ESI-MS (m/z): [ M + H ]] ⁺ ，552。

Synthesis of the compound N- (2-chloro-4, 6-difluorophenyl) -2- ((6- (4- (2-hydroxyethyl) piperazin-1-yl) -2-methylpyrimidin-4-yl) amino) thiazole-5-carboxamide

LiOH (150 mg) was added to the compound 2- (4- (6- ((5- ((2-chloro-4, 6-difluorophenyl) carbamoyl) thiazole-2-yl) amino) -2-methylpyrimidin-4-yl) piperazin-1-yl) ethyl acetate (150mg, 0.272mmol) in a mixture of methanol/water (5/1 mL) was stirred at room temperature for 30 minutes (reaction monitored by LCMS) and concentrated in vacuo to give the compound N- (2-chloro-4, 6-difluorophenyl) -2- ((6- (4- (2-hydroxyethyl) piperazin-1-yl) -2-methylpyrimidin-4-yl) amino) thiazole-5-carboxamide, which was used directly in the next reaction. The purity is 99.4%. ESI-MS (m/z): [ M + H ]] ⁺ ，510。

Synthesis of the compound 4- (6- ((5- ((2-chloro-4, 6-difluorophenyl) carbamoyl) thiazol-2-yl) amino) -2-methylpyrimidin-4-yl) -1- (2-hydroxyethyl) piperazine 1-oxide

H is to be ₂ O ₂ (30% strength, 2 ml) was added to a solution of the compound N- (2-chloro-4, 6-difluorophenyl) -2- ((6- (4- (2-hydroxyethyl) piperazin-1-yl) -2-methylpyrimidin-4-yl) amino) thiazole-5-carboxamide (138mg, 0.272mmol) in NMP (2 ml), stirred at room temperature for 24 hours, purified by high performance liquid chromatography (eluent system water/acetonitrile), concentrated and lyophilized to give 80mg as a white solid in free form in 56% yield over two steps, with a purity of 99%. ESI-MS (m/z): [ M + H ]] ⁺ ，526。 ¹ H NMR(400MHz，DMSO-d ₆ ) δ:2.43 (s, 3H, pyrimidine ring-CH) ₃ )，3.25-3.33(m，8H，CH ₂ )，3.67-3.61(t，J＝12Hz，2H)，3.91(s，2H，CH ₂ ) 4.12 (2H, active hydrogen), 6.16 (s, 1H, pyrimidinehydrogen), 7.52-7.47 (m, 2H, aromatic hydrogen), 8.22 (s, 1H, thiazolehydrogen).

Preparation example 5

Synthesis of Compound 5:

the synthetic route for compound 5 is as follows:

2- ((6- (4- (2-acetoxyethyl) piperazin-1-yl) -2-methylpyrimidin-4-yl) amino) thiazole-5-carboxylic acid was prepared according to reaction steps 1-4 of compound 2.

Synthesis of the compound 2- (4- (6- ((5- ((2, 6-dichloro-4-fluorophenyl) carbamoyl) thiazol-2-yl) amino) -2-methylpyrimidin-4-yl) piperazin-1-yl) ethyl acetate

The compound 2- ((6- (4- (2-acetoxyethyl) piperazin-1-yl) -2-methylpyrimidin-4-yl) amino) thiazole-5-carboxylic acid (1.0g, 2.462mmol) and the compound 2, 6-dichloro-4-fluoroaniline (485mg, 2.708mmol) were added to a pyridine (5 mL) solution, and POCl was added dropwise at 0 ℃ in an ice bath ₃ (0.5 mL), stir at room temperature for 60 minutes, after the reaction was complete (reaction monitored by LCMS), add ethyl acetate (15 mL) and water (10 mL), saturated NaHCO ₃ The aqueous solution was adjusted to PH =6 to 7, the aqueous phase was extracted with ethyl acetate (15 mL × 3), the organic phase was washed with water (25 mL × 3) and saturated brine (25 mL × 1), respectively, dried over anhydrous sodium sulfate, filtered, and concentrated to give a crude product as a brown oil, which was purified by high performance liquid chromatography (mobile phase acetonitrile/water), concentrated and lyophilized to give 200mg of a yellow solid, yield 14.1%, purity 96.3%. ESI-MS (m/z): [ M + H ]] ⁺ ，568。

Synthesis of the compound N- (2, 6-dichloro-4-fluorophenyl) -2- ((6- (4- (2-hydroxyethyl) piperazin-1-yl) -2-methylpyrimidin-4-yl) amino) thiazole-5-carboxamide

LiOH (200 mg) was added to a methanol/water mixture (5/1 mL) of compound 2- (4- (6- ((5- ((2, 6-dichloro-4-fluorophenyl) carbamoyl) thiazol-2-yl) amino) -2-methylpyrimidin-4-yl) piperazin-1-yl) ethyl acetate (200mg, 0.353mmol), stirred at room temperature for 30 minutes (LCMS monitoring reaction), and concentrated in vacuo to give compound N- (2, 6-dichloro-4-fluorophenyl) -2- ((6- (4- (2-hydroxyethyl) piperazin-1-yl) -2-methylpyrimidin-4-yl) amino) thiazole-5-carboxamide, which was used directly in the next reaction. The purity is 92.1%. ESI-MS (m/z): [ M + H ]] ⁺ ，526。

Synthesis of the compound 4- (6- ((5- ((2, 6-dichloro-4-fluorophenyl) carbamoyl) thiazol-2-yl) amino) -2-methylpyrimidin-4-yl) -1- (2-hydroxyethyl) piperazine 1-oxide

Will H ₂ O ₂ (30% strength, 2 ml) was added to a solution of the compound N- (2, 6-dichloro-4-fluorophenyl) -2- ((6- (4- (2-hydroxyethyl) piperazin-1-yl) -2-methylpyrimidin-4-yl) amino) thiazole-5-carboxamide (165mg, 0.353mmol) in the previous step in NMP (2 ml), stirred at room temperature for 24 hours, purified by high performance liquid chromatography (eluent system water/acetonitrile), concentrated and lyophilized to give 80mg of a yellow solid, 42% yield in two steps, and 98% purity. ESI-MS (m/z): [ M + H ]] ⁺ ，542。 ¹ H NMR(400MHz，DMSO-d ₆ ) δ:2.43 (s, 3H, pyrimidine ring-CH ₃ )，3.25-3.36(m，8H，CH ₂ )，3.61-3.67(t，J＝12Hz，2H)，3.91(s，2H，CH ₂ ) 4.09-4.11 (2H, active hydrogen), 6.13 (s, 1H, pyrimidine hydrogen), 7.64-7.66 (m, 2H, aromatic hydrogen), 8.21 (s, 1H, thiazole hydrogen).

Example 1

The compound of the formula I is adopted to carry out in-vitro kinase inhibition activity research and human liver microsome metabolism research. 1. Study of kinase inhibitory Activity of Compound 1

1.1 Experimental methods

1.1.1 test drug formulation

Experimental groups: compound 1 and dasatinib were dissolved in 50 μ l DMSO, respectively, at a final drug concentration of 10mM.

Positive control group: staurosporine (Med Chem as a supplier) was used and DMSO as a matrix solution.

1.1.2 doses administered

Experimental group initial concentration of drug: 1 μ M

Initial concentration of positive control group: 20 μ M

Incubation time of compound: 15min

ATP concentration: 10 μ M

Reaction time: 2h

The reaction doses (M) are shown in the table below:

1.1.3 Experimental conditions and procedures

Buffer conditions: 20mM Hepes (pH 7.5), 10mM MgCl ₂ ，1mM EGTA，0.02％Brij35，0.02mg/ml BSA，0.1mM Na ₃ VO ₄ ，2mM DTT，1％DMSO

Remarking: the required coenzyme factors are added separately to each kinase reaction.

The measurement conditions were as follows:

the reaction process is as follows:

a) The indicator substrate was prepared in a freshly prepared reaction buffer.

b) Any necessary coenzyme factors are added to the above substrate solution.

c) The indicator kinase was added to the substrate solution and gently mixed.

d) The test drug was added to the kinase reaction mixture using Echo 550.

e) Will be provided with ³³ P-ATP (radioactivity 0.01. Mu. Ci/. Mu.l) was added to the reaction mixture and the reaction was initiated.

f) The kinase reaction was incubated at room temperature for 120 minutes.

g) The reaction showed spots on P81 ion exchange paper.

h) The filter was cleaned using 0.75% phosphoric acid.

i) The residual radiophosphorylated substrate on the filter paper was measured.

1.2 data analysis

Data sheet for kinase ActivityIs shown as the percentage of kinase activity remaining in the test sample that reacted with the carrier (dimethyl sulfoxide). IC was obtained using Prism4 software (GraphPad) ₅₀ Values and curve fitting.

1.3 results of the experiment

1.4 conclusion of the experiment

The compounds of the present disclosure, e.g., compound 1, inhibited ABL1, BTK (C481S) activity comparable to or higher than dasatinib, significantly higher than the control group.

2. Study of kinase inhibitory Activity of Compound 2 and Compound 5

2.1 Experimental methods

2.1.1 test drug formulation

Experimental groups: test compounds (compound 2 and compound 5) were dissolved in DMSO, respectively, at a final drug concentration of 10mM.

Compound (I)	DMSO(uL)
		Compound 2	302.3
Compound 5	47.3

Positive control group: staurosporine (LC Laboratories, supplier) was used and the matrix solution was DMSO.

2.1.2 doses administered

Experimental group initial concentration of drug: 1 μ M

Initial concentration of positive control group: 1 μ M

Compound incubation time: for 10min

ATP concentration: 20 μ M (BTK), 12 μ M (BTK-C481S), 14 μ M (ABL 1)

Reaction time: 90min (BTK/BTK-C481S), 60min (ABL 1)

The reaction doses (M) are shown in the table below:

2.1.3 Experimental conditions and procedures

Buffer conditions (ABL 1): 50mM Hepes (pH 7.5), 10mM MgCl ₂ ，1mM EDTA，0.01％Brij35。

Buffer conditions (BTK/BTK-C481S): 50mM Hepes (pH 7.5), 10mM MgCl ₂ ，1mM EDTA，0.01％Brij35，1mM DTT。

The measurement conditions were as follows:

kinase classes	Kinase amounts in RXN	Suppliers of goods
			ABL1	0.2nM	Invitrogen
BTK	1nM	Carna
			BTK-C481S	0.5nM	Carna

Substrate	Amount of substrate in RXN	Suppliers of goods
			ABL1 substrates	2μM	Invitrogen
BTK substrates	1μM	Cisbio
			BTK-C481S substrate	1μM	Cisbio

The reaction process comprises the following steps:

a) Test compounds and positive compounds were injected into the assay plates using Echo 550.

b) The enzyme was mixed with the corresponding substrate and added to the assay plate. Incubate at 23 ℃ for 10 minutes.

c) The reaction was initiated by the addition of ATP. The reaction time was 90 minutes (BTK/BTK-C481S), 60 minutes (ABL 1).

d) The reaction was further developed by adding the relevant reagents. Time 60min (BTK/BTK-C481S), 40 min (ABL 1).

e) Detection was performed on Envision.

2.2 data analysis

For BTK/BTK-C481S: data calculation is carried out by using Model205 in XLFit5 software to obtain IC ₅₀ The value is obtained. For ABL1: firstly converting the original signal into phosphorylation rate, then converting the phosphorylation rate into inhibition rate, and then calculating IC by using XLFit5 software ₅₀ 。

2.3 results of the experiment

3. Liver microsome metabolism test

3.1 Experimental methods

3.1.1 incubation conditions

Substrate concentration: 1.0. Mu.M

Buffer solution: 0.05M phosphate buffer (pH 7.4)

Human liver microsomes:

UltraPool ^TM HLM 150，20mg/mL

microparticle protein concentration: 0.25mg/mL

NADPH concentration: 1.0mM

Total reaction volume: 850 μ L

Incubation temperature: 37 deg.C

Pre-incubation time: 5min (without NADPH)

Sampling time: 0,5, 10, 15, 20, 30min (control group is 0, 15, 30 min)

Sampling volume: samples 85 μ L per time point

Quenching reagent: 85 u L acetonitrile containing 0.25 u M warfarin

Test compounds: verapamil, dasatinib and compound 1

3.1.2 Experimental design

3.1.3 Experimental procedures

a) Acetonitrile is adopted: water (50.

b) Microsomal protein was diluted to 0.5mg/mL with 50mM phosphate buffered saline and placed in an ice bath (human liver microsome concentration 20 mg/mL).

c) A2.5 mM NADPH solution was prepared.

d) Liver microsomal protein, buffer, test compound were sequentially transferred to 1.5ml incubation tubes according to the volumes in the experimental design table, incubated at 37 ℃ for 5min with constant shaking.

e) Add 85. Mu.L of quenching reagent in a 250. Mu.L quench vial and place on an ice bath.

f) The reaction was started by addition of NADPH, 85 μ L was sampled immediately and controlled incubation and quenching in quench vials mixed well as a 0min sample point. The samples were capped, vortexed and centrifuged at 4 ℃.

g) The quenched solution was left at 4 ℃ for 10min for precipitation of the protein.

h) The operation of the other sampling points is similar to the 0min sampling operation.

i) The sample quenching reaction mixture solution was thoroughly mixed well and centrifuged at 14000rpm for 10min at room temperature.

j) The supernatant (. About.100. Mu.l) was removed for LC-MS analysis.

3.2 LC-MS analysis method

The instrument comprises the following steps:

ACQUITY HPLC-Xevo G2-XS QTof

a chromatographic column: ACQUITY

BEH C ₁₈ Column 2.1X 50mm,1.7 μm

Mobile phase: mobile phase a was water containing 0.1% formic acid and mobile phase B was acetonitrile containing 0.1% formic acid. The gradient is a gradient of 5 to 95% by weight B every 5.1 min.

Flow rate: 100 μ l/min.

The amount of parent compound per time was determined from the peak area ratio (compound area/warfarin area).

3.3 calculation formula

Intrinsic Clearance (CL) _int ) Calculated by the following formula.

Clearance rate constant (k) = -slope (1/min)

Half life (t) _1/2 )＝0.693/k

V (μ L/mg) = incubation volume (μ L)/amount of incubation protein (mg)

Intrinsic Clearance (CL) _int )(μL/min/mg)＝V×0.693/t _1/2

3.4 results of the experiment

3.5 conclusion of the experiment

The compound of formula I disclosed by the invention is stable in vitro metabolism, has a longer half-life period and a lower clearance compared with dasatinib, and can prolong the administration interval and reduce the administration frequency.

The invention has been described in detail with respect to a general description and specific embodiments thereof, but it will be apparent to those skilled in the art that modifications and improvements can be made based on the invention. Accordingly, it is intended that all such modifications and alterations be included within the scope of this invention as defined in the appended claims.

Claims

1. A compound of formula I, or a pharmaceutically acceptable salt or stereoisomer thereof,

wherein:

R ₀ selected from halogens;

R ₂ 、R ₃ 、R ₅ 、R ₇ 、R ₁₀ 、R ₁₁ 、R ₁₂ 、R ₁₃ 、R ₁₄ 、R ₁₅ 、R ₁₆ 、R ₁₇ each independently selected from H;

R ₁ is selected from C _1-6 An alkyl group;

R ₄ selected from halogen;

R ₆ each independently selected from C _1-6 An alkyl group; r ₈ Is selected from C _1-6 A hydroxyalkyl group.

2. A compound of formula I according to claim 1, or a pharmaceutically acceptable salt or stereoisomer thereof, wherein:

R ₀ selected from fluorine, chlorine, bromine and iodine.

3. A compound of formula I according to claim 1 or 2, or a pharmaceutically acceptable salt or stereoisomer thereof, wherein:

R ₁ is selected from C _1-4 An alkyl group;

R ₄ selected from halogens; and is

R ₆ Is selected from C _1-4 An alkyl group.

4. A compound of formula I according to claim 1 or 2, or a pharmaceutically acceptable salt or stereoisomer thereof, wherein:

R ₀ selected from fluorine.

5. A compound of formula I according to claim 1 or 2, or a pharmaceutically acceptable salt or stereoisomer thereof, wherein:

R ₁ selected from methyl and isopropyl.

6. A compound of formula I according to claim 1 or 2, or a pharmaceutically acceptable salt or stereoisomer thereof, wherein:

R ₄ is chlorine.

7. A compound of formula I according to claim 1 or 2, or a pharmaceutically acceptable salt or stereoisomer thereof, wherein:

R ₆ selected from methyl.

8. A compound of formula I according to claim 1 or 2, or a pharmaceutically acceptable salt or stereoisomer thereof, wherein:

R ₈ is selected from C _1-4 A hydroxyalkyl group.

9. A compound of formula I as claimed in claim 1 or 2, or a pharmaceutically acceptable salt or stereoisomer thereof, wherein:

R ₈ selected from hydroxyethyl groups.

10. The following compounds, or a pharmaceutically acceptable salt or stereoisomer thereof,

11. a pharmaceutical composition comprising a compound of any one of claims 1-10, or a pharmaceutically acceptable salt or stereoisomer thereof.

12. The pharmaceutical composition of claim 11, further comprising a pharmaceutically acceptable carrier, excipient, or adjuvant.

13. The pharmaceutical composition of claim 11 or 12, further comprising another therapeutic agent for combination therapy.

14. The pharmaceutical composition of claim 13, wherein the additional therapeutic agent is selected from one or more of the following: cyclophosphamide, ifosfamide, vincristine, daunorubicin, doxorubicin, cytarabine, mitoxantrone, dacarbazine, idarubicin, retinoic acid, prednisone, dexamethasone, mercaptopurine, methotrexate, paclitaxel, melphalan, long-acting interferon, venetocel, crizotinib, erlotinib, ocitinib, ruxotinib, afatinib, erlotinib, imatinib, lapatinib, bevacizumab, trastuzumab, rituximab, cetuximab, bornatuzumab, fludarabine, gemcitabine, decitabine, capecitabine, bendamustine, everolimus, temsirolimus, etoposide, granulocyte colony stimulating factor, temozolomide, zoledronic acid, olixaplatin, cisplatin, carboplatin, and fulvestrant.

15. Use of a compound according to any one of claims 1 to 10, or a pharmaceutically acceptable salt or stereoisomer thereof, or a pharmaceutical composition according to any one of claims 11 to 14, in the manufacture of a medicament for the prevention or treatment of a tyrosine kinase-associated disease.

16. Use of a compound of any one of claims 1-10, or a pharmaceutically acceptable salt or stereoisomer thereof, or a pharmaceutical composition of any one of claims 11-14, in the manufacture of a medicament for inhibiting tyrosine kinase activity.

17. The use of claim 15, wherein:

the tyrosine kinase-associated diseases are diseases, disorders, and conditions that benefit from inhibition or reduction of tyrosine kinase activity.

18. The use of any one of claims 15-17, wherein:

the tyrosine kinases include Bcr-Abl tyrosine kinase and BTK tyrosine kinase.

19. The use of claim 17, wherein:

the disease, disorder or condition is selected from cancer;

the cancer is selected from: chronic Myelogenous Leukemia (CML), gastrointestinal stromal tumor (GIST), small Cell Lung Cancer (SCLC), non-small cell lung cancer (NSCLC), multiple myeloma, solid tumor, B-cell lymphoma, chronic Lymphocytic Leukemia (CLL), acute Lymphocytic Leukemia (ALL), non-hodgkin's lymphoma (NHL), small Lymphocytic Lymphoma (SLL), mantle Cell Lymphoma (MCL), melanoma, mastocytosis, germ cell tumor, acute Myeloid Leukemia (AML), marginal zone/diffuse large B cell lymphoma, sarcoma, pancreatic cancer, glioblastoma, head and neck tumors, macroglobulinemia, follicular central lymphoma, prostate cancer, myelodysplastic syndrome, atherosclerotic myelohyperplasia, myelofibrosis, eosinophilia, polycythemia vera, liver cancer, advanced sarcoma, glioblastoma multiforme, gliosarcoma, malignant mesothelioma, melanoma, head and neck cell carcinoma, skin carcinoma, neuroendocrine tumors, B cell lymphoma, gastric cell lymphoma, acute lymphoblastic leukemia, metastatic lymphoma of small cell lymphoma, metastatic carcinoma of the rectum, metastatic carcinoma of the head and neck, metastatic carcinoma of the bladder, metastatic carcinoma of small cell lymphoma, metastatic carcinoma of the prostate and neck.

20. The use of claim 17, wherein:

the disease, disorder or condition is selected from the group consisting of cancer with chemotherapeutic agent resistance to the target BCR-ABL and c-KIT, and cancer with resistance to imatinib.

21. The use of claim 17, wherein:

the diseases, disorders and conditions are selected from: bone metastasis, hypercalcemia and/or osteoporosis; pulmonary fiber disease; cardiovascular disease or condition; mast cell mediated inflammatory diseases; HTLV-1 associated myelopathy/tropical spastic paralysis; complex Regional Pain Syndrome (CRPS); weight loss or fat loss; arterial occlusive disease; ubiquitination; diseases or conditions associated with reduced function of degrading sugars; fridureich ataxia; graft rejection in parkinson's disease progression; rheumatoid arthritis; graft versus host disease; (ii) an autoimmune disease; recurrent immune thrombocytopenic purpura, pemphigus vulgaris, systemic lupus erythematosus, scleroderma pulmonary interstitial fibrosis and idiopathic urticaria.

22. The use of claim 21, wherein:

the cardiovascular disease or condition is a cardiovascular disease caused by RASopathy, or a congenital heart disease associated with nosan or nosan syndrome.

23. The use of claim 21, wherein:

the mast cell mediated inflammatory disease is selected from osteoarthritis, asthma, chronic obstructive pulmonary disease uveitis, aspirin-aggravated respiratory disease (AERD), and parkinson's disease.