CN109206435B

CN109206435B - Thieno [3,2-d ] pyrimidine compound and preparation method and medical application thereof

Info

Publication number: CN109206435B
Application number: CN201710513037.8A
Authority: CN
Inventors: 殷惠军; 田卫学; 闫旭; 宗利斌; 陈牛艳; 米帧; 伍洲; 马静; 李英辉; 王洪志; 杨琰
Original assignee: National Institutes of Pharmaceutical R&D Co Ltd
Current assignee: National Institutes of Pharmaceutical R&D Co Ltd
Priority date: 2017-06-29
Filing date: 2017-06-29
Publication date: 2020-09-08
Anticipated expiration: 2037-06-29
Also published as: CN109206435A

Abstract

The invention relates to thieno [3,2-d ]]Pyrimidine compounds, and preparation methods and medical uses thereof. In particular, the invention relates to a compound shown in a general formula I, a preparation method thereof, a pharmaceutical composition containing the compound, and application of the compound as a BTK kinase inhibitor, wherein the compound and the pharmaceutical composition containing the compound can be used for treating diseases related to BTK kinase activity, such as inflammation, autoimmune diseases, cancer and the like. Wherein the definition of each substituent in the general formula I is the same as that in the specification.

Description

Thieno [3,2-d ] pyrimidine compound and preparation method and medical application thereof

Technical Field

The invention belongs to the technical field of medicines, and particularly relates to a thieno [3,2-d ] pyrimidine compound, a preparation method thereof, a pharmaceutical composition containing the same, and application thereof in regulating Bruton kinase (BTK) activity and treating and/or preventing diseases related to BTK activity.

Background

Intracellular signaling processes are an efficient way for cells to respond to external stimuli and ultimately elicit specific biological effects. Cytokines are capable of intracellular signaling through a variety of signal transduction pathways, thereby participating in the regulation of cell proliferation, hematopoietic functions and many important biological functions associated with immunity. Bruton kinase (BTK) among protein tyrosine kinases plays an important role in cytokine signal transduction.

The BTK family is a class of non-receptor tyrosine protein kinases. In 1952, Bruton found that X-linked agammaglobulinemia (XLA) patients lack mature B cells and cannot effectively generate autonomous immune response, and gene research found that X chromosome abnormality of the patients causes BTK gene mutation, which indicates that BTK has an important role in the developmental maturity of the B cells. BTK is not only regulated by the B cell antigen receptor (BCR) pathway to participate in immune responses, but also regulated by the Toll-like receptor (TLR) signaling pathway to participate in inflammatory responses.

BTK family members are mainly distributed in hematopoietic tissues in human bodies, and have important regulation effects on growth and differentiation of hematopoietic cells. Among them, BTK kinase is mainly distributed in bone marrow-derived cells and B cells. BTK comprises 5 domains: the PH domain, responsible for mediating translocation of proteins to the cell membrane; a TH region containing two modules of PRR and Btk; SH2 and SH3 play an important role in mediating the interaction between BTK and other protein molecules; and kinase catalytic domain SH 1.

BTK is activated after being combined with PI3 (phosphatidylinositol-3-kinase K), G-protein dual receptor and the like, and participates in regulation of angiogenesis, cell proliferation and apoptosis and cell movement through downstream signal molecules such as PLC gamma 2 (phospholipase C-gamma 2) and PKC beta I (serine threonine kinase beta I). During BTK activation, the PH region binds to phosphatidylinositol-3, 4, 5-trisphosphate (PIP3), completing membrane translocation. BTK after membrane localization is activated and involved in the phosphate group transfer of the two tyrosine residues Tyr551 and Tyr223, the Tyr551 residue on the activation loop is phosphorylated by Src family tyrosine kinases, resulting in the autophosphorylation of Tyr233 in the SH3 domain, providing the necessary conditions for the complete activation of the kinase. BTK activation is negatively fed back and regulated by PKC β, which phosphorylates Ser180 of BTK, reducing its recruitment and phosphate group transfer to the cell membrane, thereby reducing its activity. PKC inhibitory signals have a key role in maintaining a balance of BTK activity in the B cell receptor signaling pathway.

BTK plays an important role in the development of B cell malignancies through the BCR and TCR pathways. The BTK inhibitor shows good treatment effect by taking BTK function as a target spot in various B cell malignant tumors, and a plurality of BTK inhibitors show exact treatment effect in clinical experiments aiming at chronic lymphocytic leukemia, mantle cell lymphoma, diffuse large B cell lymphoma and multiple myeloma. Moreover, the role of BTK inhibitors in other B-cell malignancies has also attracted interest, such as the discovery of a MYD88 mutation and spleen marginal zone lymphoma in which the NF- κ B signaling pathway is involved, as well as follicular lymphoma in which BCR antigen (and autoantigen) recognition is implicated in cell survival. It is noteworthy that the incidence of diffuse large B-cell lymphoma and marginal zone lymphoma is higher in patients with systemic autoimmune diseases, whereas activation of BTK pathway is an important pathological event of autoimmune diseases, and BTK inhibitors show stronger effects in preclinical experiments. BTK has also been shown to protect breast cancer cells from apoptosis.

Rheumatoid Arthritis (RA) is an autoimmune disease characterized by inflammation and destruction of joint structures. When the disease is not treated effectively, substantial disability and pain, and even premature death, result from loss of joint functionality. The aim of RA treatment is therefore not only to delay the progression of the disease but also to obtain a reduction in symptoms, thereby terminating joint destruction. The global prevalence of RA is about 0.8%, with women having a three-fold prevalence rate over men. RA is difficult to treat, there is currently no cure, and treatment focuses on relieving pain and preventing diseased joint degeneration. Clinical treatment strategies include nonsteroidal anti-inflammatory drugs (NSAIDs), hormones, disease-modifying antirheumatic drugs (DMARDS), and biologic drugs, mainly to relieve the symptoms of joint damage and swelling. Clinical application of DMARDS (such as methotrexate, hydroxychloroquine, leflunomide, sulfasalazine) and DMARDS has better effect when being combined with biological drugs. Despite the abundance of anti-RA drugs, pain still exists in more than 30% of patients. Recent studies have shown that intervention of the BTK signaling pathway is a new approach to RA treatment.

T cells remain attractive targets in the treatment of RA due to their abundant presence in the synovial cavity and their ability to activate B cells in RA. Although blocking therapy for T cell co-stimulation has been licensed, depletion of T cells in RA therapy has shown limited therapeutic value. In addition, B cell function and autoantigen production have been demonstrated during disease progression, and thus one has targeted B cells directly or indirectly for therapy. Importantly, therapies aimed at reducing B cells effectively improved the clinical symptoms of RA, suggesting that B cells play an important role in RA disease. The effectiveness of B cell-targeted therapies (either to reduce B cell numbers or inhibit B cell survival) in RA demonstrates the important role of B cells in RA. BTK plays an important role in B cell maturation and activation by participating in the BCR signaling pathway. The BCR pathway is enhanced in BTK-highly expressed B cells and causes the overexpression of autoantibodies, thereby triggering an autoimmune response. Thus, BTK is an ideal target for therapy in the treatment of autoimmune diseases including, but not limited to, rheumatoid arthritis, multiple sclerosis, lupus, psoriasis, psoriatic arthritis, inflammatory bowel disease, crohn's disease, uveitis, sarcoidosis. Several BTK inhibitors have shown good therapeutic efficacy in the treatment of autoimmune diseases in preclinical animal models.

In recent years, BTK inhibitors have progressed faster. Ibrutinib is mainly used for treating B cell lymphoma, and CC-292 and ACP-196 in the middle and later clinical stages are used for treating B cell lymphoma and also used for developing autoimmune disease indications such as RA. However, these BTK inhibitors generally have the disadvantages of low activity and selectivity, and large side effects. Thus, there is a continuing need for new or improved agents that inhibit kinases such as Bruton's kinase for the development of new, more potent drugs to treat RA or other BTK-related diseases.

Disclosure of Invention

The inventor designs and synthesizes a series of compounds containing thieno [3,2-d ] pyrimidine skeleton through intensive research, screens the BTK kinase activity of the compounds, and shows that the compounds have outstanding activity of resisting the BTK kinase and can be developed into medicines for treating diseases related to the BTK kinase activity.

Therefore, the invention aims to provide a compound shown in the general formula I or a racemate, an enantiomer, a diastereoisomer, a mixture form, a prodrug or a pharmaceutically acceptable salt thereof,

wherein:

x is selected from O or NH;

R¹selected from the group consisting of hydrogen, halogen, amino, nitro, cyano, hydroxy, mercapto, carboxyl, ester, alkyl, haloalkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl;

y is selected from CH₂O, S or NR²；

R²Selected from hydrogen, alkyl, cycloalkyl; wherein said alkyl, cycloalkyl is optionally further substituted with one or more groups selected from halogen, amino, nitro, cyano, oxo, hydroxy, mercapto, carboxyl, ester, alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl;

z is

R³And R⁴Each independently selected from hydrogen, alkyl, wherein said alkyl is optionally further selected from heterocyclyl, -NR^aR^bSaid heterocyclyl is optionally further substituted by halogen or alkyl;

w is selected from NH or O;

cy is selected from aryl, heteroaryl, wherein said aryl, heteroaryl are optionally further substituted with one or more groups selected from hydrogen, halogen, alkyl, alkoxy, haloalkyl, haloalkoxy;

L¹and L²Each independently selected from the group consisting of a single bond, - (CH)₂)_m-、-O-、-NR⁵-、-S-、-SO-、-SO₂-and-CO-one or more;

m is an integer of 1 to 4;

R⁵selected from hydrogen, alkyl, cycloalkyl; wherein said alkyl, cycloalkyl is optionally further substituted with one or more groups selected from halogen, amino, nitro, cyano, oxo, hydroxy, mercapto, carboxyl, ester, alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl;

R⁶selected from alkyl, cycloalkyl, heterocyclyl; wherein said alkyl, cycloalkyl, heterocyclyl is optionally further substituted with one or more R⁷Substitution;

each R⁷Each independently selected from halogen, amino, nitro, cyano, hydroxy, mercapto, or a salt thereof,Oxo, alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl, -C (O) R^a、-O(O)CR^a、-C(O)OR^a、-C(O)NR^aR^b、-NHC(O)R^a、-S(O)R^a、-S(O)₂R^a、-S(O)NR^aR^b、-NR^aR^b、-S(O)₂NR^aR^b、-NHS(O)R^a、-NHS(O)₂R^a(ii) a Wherein said alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl are optionally further substituted with one or more groups selected from halogen, amino, nitro, cyano, oxo, hydroxy, mercapto, carboxyl, ester, alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl;

R^aand R^bEach independently selected from hydrogen, halogen, hydroxy, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, wherein said alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl are optionally further substituted with one or more groups selected from halogen, amino, nitro, cyano, hydroxy, mercapto, carboxyl, ester, oxo, alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl;

or R^aAnd R^bTogether with the nitrogen atom to which they are attached form a nitrogen-containing heterocyclic group that is optionally further substituted with one or more groups selected from halogen, amino, nitro, cyano, oxo, hydroxy, mercapto, carboxyl, ester, alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclic, aryl, heteroaryl.

In a preferred embodiment of the present invention, the compound of formula I according to the present invention, or its racemate, enantiomer, diastereomer, or mixture thereof, or its prodrug, or its pharmaceutically acceptable salt, is a compound of formula II, or its racemate, enantiomer, diastereomer, or mixture thereof, or its prodrug, or its pharmaceutically acceptable salt,

wherein R is¹、R⁶、Y、Z、Cy、L¹、L²As defined in formula I.

In another preferred embodiment of the present invention, the compound of formula I according to the present invention, or its racemate, enantiomer, diastereomer, or mixture thereof, its prodrug, or its pharmaceutically acceptable salt, is a compound of formula III, or its racemate, enantiomer, diastereomer, or mixture thereof, its prodrug, or its pharmaceutically acceptable salt,

wherein R is¹、R⁶、Y、Z、Cy、L¹、L²As defined in formula I.

In another preferred embodiment of the present invention, the compound of formula I according to the present invention, or its racemate, enantiomer, diastereomer, or mixture thereof, its prodrug, or its pharmaceutically acceptable salt, is a compound of formula IV, or its racemate, enantiomer, diastereomer, or mixture thereof, its prodrug, or its pharmaceutically acceptable salt,

wherein R is¹、R⁶、Y、Z、Cy、L¹、L²As defined in formula I.

In another preferred embodiment of the present invention, the compound of formula I according to the present invention, or its racemate, enantiomer, diastereomer, or mixture thereof, prodrug thereof, or pharmaceutically acceptable salt thereof, wherein Cy is aryl, preferably phenyl.

In another preferred embodiment of the present invention, the compound of formula I according to the present invention, or its racemate, enantiomer, diastereomer, or mixture thereof, prodrug thereof or pharmaceutically acceptable salt thereof, wherein Y is selected from CH₂Or O.

In another preferred embodiment of the present invention, the compound of formula I according to the present invention, or its racemate, enantiomer, diastereomer, or mixture thereof, prodrug thereof or pharmaceutically acceptable salt thereof, wherein Z is

R³And R⁴Each independently selected from hydrogen or alkyl.

R³Selected from hydrogen or alkyl.

In another preferred embodiment of the present invention, the compound of formula I according to the present invention or its racemate, enantiomer, diastereomer, or mixture thereof, prodrug thereof or pharmaceutically acceptable salt thereof, wherein L is¹And L²Each independently selected from the group consisting of a single bond, - (CH)₂)_mOne or more of-, -O-, and-CO-; m is an integer of 1 to 4.

In another preferred embodiment of the present invention, the compound of formula I according to the present invention, or its racemate, enantiomer, diastereomer, or mixture thereof, prodrug thereof, or pharmaceutically acceptable salt thereof, wherein R is⁶Is selected from C₁-C₆Alkyl or C₃-C₇A cycloalkyl group; wherein said alkyl and cycloalkyl groups are optionally further substituted by one or more R⁷Substitution; each R⁷Each independently selected from halogen, amino, nitro, cyano, hydroxy, mercapto, oxo, C₁-C₆Alkyl radical, C₁-C₆Alkoxy radical, C₃-C₇Cycloalkyl, 5-to 7-membered heterocyclyl, C₅-C₁₀Aryl, 5 to 10 membered heteroaryl.

In another preferred embodiment of the present invention, the compound of formula I according to the present invention, or its racemate, enantiomer, diastereomer, or mixture thereof, prodrug thereof, or pharmaceutically acceptable salt thereof, wherein R is⁶Selected from 5-to 7-membered heterocyclyl, preferably piperazinyl, morpholinyl, piperidinyl, thiomorpholinyl, wherein said heterocyclyl is optionally further substituted with one or more R⁷Substitution; each R⁷Each independently selected from halogen, amino, nitro, cyano, hydroxy, mercapto, oxo, C₁-C₆Alkyl radical, C₁-C₆Alkoxy radical, C₃-C₇Cycloalkyl, 5-to 7-membered heterocyclyl, C₅-C₁₀Aryl, 5 to 10 membered heteroaryl.

Typical compounds of the invention include, but are not limited to:

or a racemate, enantiomer, diastereomer, or mixture thereof, a prodrug thereof, or a pharmaceutically acceptable salt thereof

In another aspect, the present invention provides a method for preparing a compound of formula I according to the present invention, or its racemate, enantiomer, diastereomer, or mixture thereof, its prodrug, or its pharmaceutically acceptable salt, the method comprising the steps of:

reacting the compound Id with a compound H-W-Cy-L under high-temperature acidic conditions¹-L²-R⁶Reacting to obtain a compound shown in the general formula I, wherein the acid reagent is preferably trifluoroacetic acid, and the temperature is preferably 100 ℃;

wherein W, X, Y, Z, Cy, L¹、L²、R¹、R⁶As defined in formula I.

Another aspect of the present invention relates to a pharmaceutical composition, which comprises an effective amount of a compound represented by formula I, or its racemate, enantiomer, diastereomer, or mixture thereof, its prodrug, or its pharmaceutically acceptable salt, and a pharmaceutically acceptable carrier, diluent, or excipient. The invention also relates to a method for preparing the composition, which comprises the step of mixing the compound shown in the general formula I or the raceme, the enantiomer, the diastereoisomer, the mixture form, the prodrug or the pharmaceutically acceptable salt thereof with a pharmaceutically acceptable carrier, diluent or excipient.

The invention further relates to a compound shown in the general formula I, or a racemate, an enantiomer, a diastereoisomer, a mixture form, a prodrug or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing the compound and the prodrug, and application of the compound in preparing BTK kinase inhibitors.

The invention further relates to a compound shown in the general formula I or a racemate, an enantiomer, a diastereoisomer, a mixture form, a prodrug or a pharmaceutically acceptable salt thereof or a pharmaceutical composition containing the compound, and application of the compound in preparation of medicines for preventing and/or treating diseases related to BTK kinase activity. The disease associated with BTK activity may be selected from inflammation, autoimmune diseases, or cancer, wherein the inflammation is for example arthritis, in particular rheumatoid arthritis, psoriatic arthritis, inflammatory bowel disease, uveitis; such autoimmune diseases as multiple sclerosis, lupus, psoriasis, sarcoidosis; such as breast cancer, cervical cancer, colon cancer, lung cancer, stomach cancer, rectal cancer, pancreatic cancer, brain cancer, skin cancer, oral cancer, prostate cancer, bone cancer, kidney cancer, ovarian cancer, bladder cancer, liver cancer, fallopian tube tumor, ovarian tumor, peritoneal tumor, melanoma, solid tumor, glioma, glioblastoma, hepatocellular carcinoma, mastoid renal tumor, head and neck tumor, leukemia, lymphoma, myeloma, and non-small cell lung cancer.

The invention also relates to a compound shown in the general formula I or raceme, enantiomer, diastereoisomer or mixture form thereof, prodrug thereof or pharmaceutically acceptable salt thereof or a pharmaceutical composition containing the same, and application thereof as a BTK kinase inhibitor.

The invention further relates to a compound shown in the general formula I or a racemate, an enantiomer, a diastereoisomer, a mixture form, a prodrug or a pharmaceutically acceptable salt thereof or a pharmaceutical composition containing the compound, and application of the compound as a medicament for preventing and/or treating diseases related to BTK activity. The disease associated with BTK activity may be selected from inflammation, autoimmune diseases, or cancer, wherein the inflammation is for example arthritis, in particular rheumatoid arthritis, psoriatic arthritis, inflammatory bowel disease, uveitis; such autoimmune diseases as multiple sclerosis, lupus, psoriasis, sarcoidosis; such as breast cancer, cervical cancer, colon cancer, lung cancer, stomach cancer, rectal cancer, pancreatic cancer, brain cancer, skin cancer, oral cancer, prostate cancer, bone cancer, kidney cancer, ovarian cancer, bladder cancer, liver cancer, fallopian tube tumor, ovarian tumor, peritoneal tumor, melanoma, solid tumor, glioma, glioblastoma, hepatocellular carcinoma, mastoid renal tumor, head and neck tumor, leukemia, lymphoma, myeloma, and non-small cell lung cancer.

The present invention further relates to a method of inhibiting BTK kinase comprising administering to a patient in need thereof a therapeutically effective dose of a compound of formula I or its racemates, enantiomers, diastereomers, or mixture thereof, prodrug thereof or a pharmaceutically acceptable salt thereof or a pharmaceutical composition comprising the same.

The present invention further relates to a method for preventing and/or treating diseases associated with BTK activity, which comprises administering a therapeutically effective dose of a compound represented by the general formula I or its racemate, enantiomer, diastereomer, or mixture thereof, prodrug thereof or pharmaceutically acceptable salt thereof, or pharmaceutical composition comprising the same, to a patient in need thereof. The disease associated with BTK activity may be selected from inflammation, autoimmune diseases, or cancer, wherein the inflammation is for example arthritis, in particular rheumatoid arthritis, psoriatic arthritis, inflammatory bowel disease, uveitis; such autoimmune diseases as multiple sclerosis, lupus, psoriasis, sarcoidosis; such as breast cancer, cervical cancer, colon cancer, lung cancer, stomach cancer, rectal cancer, pancreatic cancer, brain cancer, skin cancer, oral cancer, prostate cancer, bone cancer, kidney cancer, ovarian cancer, bladder cancer, liver cancer, fallopian tube tumor, ovarian tumor, peritoneal tumor, melanoma, solid tumor, glioma, glioblastoma, hepatocellular carcinoma, mastoid renal tumor, head and neck tumor, leukemia, lymphoma, myeloma, and non-small cell lung cancer.

The compounds of formula I of the present invention may be used to form pharmaceutically acceptable acid addition salts with acids according to conventional methods in the art to which the present invention pertains. The acid includes inorganic acids and organic acids, and particularly preferably hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, benzenesulfonic acid, naphthalenedisulfonic acid, acetic acid, propionic acid, lactic acid, trifluoroacetic acid, maleic acid, citric acid, fumaric acid, oxalic acid, tartaric acid, benzoic acid, and the like.

In addition, the invention also comprises a prodrug of the compound shown in the general formula I. Prodrugs of the compounds of formula I are derivatives of the compounds of formula I which may themselves be less active or even inactive, but which, upon administration, are converted to the corresponding biologically active form under physiological conditions (e.g., by metabolism, solvolysis, or otherwise).

The pharmaceutical compositions containing the active ingredient may be in a form suitable for oral use, for example, as tablets, troches, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsions, hard or soft capsules, or syrups or elixirs. Oral compositions may be prepared according to any method known in the art for preparing pharmaceutical compositions, and such compositions may contain one or more ingredients selected from the group consisting of: sweetening agents, flavoring agents, coloring agents and preserving agents in order to provide a pleasant to the eye and palatable pharmaceutical preparation. Tablets contain the active ingredient in admixture with non-toxic pharmaceutically acceptable excipients which are suitable for the manufacture of tablets. These excipients may be inert excipients, such as calcium carbonate, sodium carbonate, lactose, calcium phosphate or sodium phosphate; granulating and disintegrating agents, such as microcrystalline cellulose, croscarmellose sodium, corn starch or alginic acid; binding agents, for example starch, gelatin, polyvinylpyrrolidone or acacia; and lubricating agents, such as magnesium stearate, stearic acid or talc. These tablets may be uncoated or they may be coated by known techniques which mask the taste of the drug or delay disintegration and absorption in the gastrointestinal tract and thereby provide a sustained action over a longer period. For example, water soluble taste masking substances such as hydroxypropylmethyl cellulose or hydroxypropyl cellulose, or time extending substances such as ethyl cellulose, cellulose acetate butyrate may be used.

Oral formulations may also be presented as hard gelatin capsules wherein the active ingredient is mixed with an inert solid diluent, for example, calcium carbonate, calcium phosphate or kaolin, or as soft gelatin capsules wherein the active ingredient is mixed with a water soluble carrier, for example polyethylene glycol, or an oil vehicle, for example peanut oil, liquid paraffin or olive oil.

Aqueous suspensions contain the active materials in admixture with excipients suitable for the manufacture of aqueous suspensions. Such excipients are suspending agents, for example sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinylpyrrolidone and acacia; dispersing or wetting agents may be a naturally occurring phosphatide, for example lecithin, or condensation products of an alkylene oxide with fatty acids, for example polyoxyethylene stearate, or condensation products of ethylene oxide with long chain aliphatic alcohols, for example heptadecaethyleneoxycetanol (heptadecaethyleneoxy cetanol), or condensation products of ethylene oxide with partial esters derived from fatty acids and a hexitol such as polyethylene oxide sorbitol monooleate, or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol anhydrides, for example polyethylene oxide sorbitan monooleate. The aqueous suspensions may also contain one or more preservatives, for example ethyl or n-propyl paraben, one or more colouring agents, one or more flavouring agents and one or more sweetening agents, such as sucrose, saccharin or aspartame.

Oil suspensions may be formulated by suspending the active ingredient in a vegetable oil, for example arachis oil, olive oil, sesame oil or coconut oil, or in a mineral oil such as liquid paraffin. The oil suspensions may contain a thickening agent, for example beeswax, hard paraffin or cetyl alcohol. Sweetening agents such as those set forth above, and flavoring agents may be added to provide a palatable preparation. These compositions can be preserved by the addition of antioxidants such as butylated hydroxyanisole or alpha-tocopherol.

Dispersible powders and granules suitable for preparation of an aqueous suspension by the addition of water may provide the active ingredient in admixture with a dispersing or wetting agent, suspending agent or one or more preservatives. Suitable dispersing or wetting agents and suspending agents are as described above. Other excipients, for example sweetening, flavoring and coloring agents, may also be present. These compositions are preserved by the addition of an antioxidant such as ascorbic acid.

The pharmaceutical compositions of the invention may also be in the form of oil-in-water emulsions. The oily phase may be a vegetable oil, for example olive oil or arachis oil, or a mineral oil, for example liquid paraffin or mixtures thereof. Suitable emulsifying agents may be naturally-occurring phosphatides, for example soy bean lecithin, and esters or partial esters derived from fatty acids and hexitol anhydrides, for example sorbitan monooleate, and condensation products of the said partial esters with ethylene oxide, for example polyethylene oxide sorbitol monooleate. The emulsions may also contain sweetening agents, flavouring agents, preservatives and antioxidants. Syrups and elixirs may be formulated with sweetening agents, for example glycerol, propylene glycol, sorbitol or sucrose. Such formulations may also contain a demulcent, a preservative, a colorant and an antioxidant.

The pharmaceutical compositions of the present invention may be in the form of a sterile injectable aqueous solution. Among the acceptable vehicles and solvents that may be employed are water, ringer's solution and isotonic sodium chloride solution. The sterile injectable preparation may be a sterile injectable oil-in-water microemulsion in which the active ingredient is dissolved in the oil phase. For example, the active ingredient is dissolved in a mixture of soybean oil and lecithin. The oil solution is then treated to form a microemulsion by adding to a mixture of water and glycerol. The injection solution or microemulsion may be injected into the bloodstream of a patient by local bulk injection. Alternatively, it may be desirable to administer the solutions and microemulsions in a manner that maintains a constant circulating concentration of the compounds of the present invention. To maintain such a constant concentration, a continuous intravenous delivery device may be used.

The pharmaceutical compositions of the present invention may be in the form of sterile injectable aqueous or oleaginous suspensions for intramuscular and subcutaneous administration. The suspension may be formulated according to the known art using those suitable dispersing or wetting agents and suspending agents which have been mentioned above. The sterile injectable preparation may also be a sterile injectable solution or suspension prepared in a non-toxic parenterally-acceptable diluent or solvent, for example as a solution in 1, 3-butanediol. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose, any blend fixed oil may be used, including synthetic mono-or diglycerides. In addition, fatty acids such as oleic acid find use in the preparation of injectables.

The compounds of the present invention may be administered in the form of suppositories for rectal administration. These pharmaceutical compositions can be prepared by mixing the drug with a suitable non-irritating excipient which is solid at ordinary temperatures but liquid in the rectum and therefore will melt in the rectum to release the drug. Such materials include cocoa butter, glycerogelatin, hydrogenated vegetable oils, polyethylene glycols of various molecular weights and mixtures of fatty acid esters of polyethylene glycols.

It is well known to those skilled in the art that the dosage of a drug administered depends on a variety of factors, including, but not limited to: the activity of the particular compound employed, the age of the patient, the weight of the patient, the health of the patient, the patient's integument, the patient's diet, the time of administration, the mode of administration, the rate of excretion, the combination of drugs, and the like. In addition, the optimal treatment regimen, such as mode of treatment, daily amount of the compound of formula (la) or type of pharmaceutically acceptable salt, can be verified according to conventional treatment protocols.

The compound containing thieno [3,2-d ] pyrimidine skeleton of the general formula I and pharmaceutically acceptable salts, hydrates or solvates thereof are used as active ingredients and mixed with a pharmaceutically acceptable carrier or excipient to prepare a composition and prepare a clinically acceptable dosage form. The derivatives of the present invention may be used in combination with other active ingredients as long as they do not produce other adverse effects such as allergic reactions and the like. The compounds of the present invention may be used as the sole active ingredient, or may be used in combination with other drugs for the treatment of diseases associated with BTK activity. Combination therapy is achieved by administering the individual therapeutic components simultaneously, separately or sequentially.

The compound has remarkable activity of regulating BTK kinase through BTK activity test, so that the compound can be used for treating and/or preventing diseases related to the activity of BTK, such as inflammation, autoimmune diseases, cancer or other diseases. In particular for the preparation of a medicament for the treatment and/or prophylaxis of rheumatoid arthritis, psoriasis and/or diseases involving cartilage and bone joint degeneration.

Detailed description of the invention

Unless stated to the contrary, terms used in the specification and claims have the following meanings.

The term "alkyl" refers to a saturated aliphatic hydrocarbon group which is a straight or branched chain group containing 1 to 20 carbon atoms, preferably an alkyl group containing 1 to 12 carbon atoms, more preferably an alkyl group containing 1 to 6 carbon atoms. Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1-dimethylpropyl, 1, 2-dimethylpropyl, 2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1, 2-trimethylpropyl, 1-dimethylbutyl, 1, 2-dimethylbutyl, 2-dimethylbutyl, 1, 3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2, 3-dimethylbutyl, n-heptyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 2, 3-dimethylpentyl, 2, 4-dimethylpentyl, 2-dimethylpentyl, 3-dimethylpentyl, 2-ethylpentyl, 3-ethylpentyl, n-octyl, 2, 3-dimethylhexyl, 2, 4-dimethylhexyl, 2, 5-dimethylhexyl, 2-dimethylhexyl, 3-dimethylhexyl, 4-dimethylhexyl, 2-ethylhexyl, 3-ethylhexyl, 4-ethylhexyl, 2-methyl-2-ethylpentyl, 2-methyl-3-ethylpentyl, n-nonyl, 2-methyl-2-ethylhexyl, 2-methyl-3-ethylhexyl, 2-dimethylpentyl, 2-dimethylhexyl, 3-dimethylpentyl, 2-ethylhexyl, 3-dimethylhexyl, 2, 2-diethylpentyl, n-decyl, 3-diethylhexyl, 2-diethylhexyl, and various branched isomers thereof. More preferred are lower alkyl groups having 1 to 6 carbon atoms, non-limiting examples of which include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1-dimethylpropyl, 1, 2-dimethylpropyl, 2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1, 2-trimethylpropyl, 1-dimethylbutyl, 1, 2-dimethylbutyl, 2-dimethylbutyl, 1, 3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2, 3-dimethylbutyl and the like. The alkyl group may be substituted or unsubstituted, and when substituted, the substituent may be substituted at any available point of attachment, preferably one or more groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halo, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, oxo, carboxy or carboxylate.

The term "alkenyl" refers to an alkyl group as defined above consisting of at least two carbon atoms and at least one carbon-carbon double bond, e.g., ethenyl, 1-propenyl, 2-propenyl, 1-, 2-or 3-butenyl, and the like. The alkenyl group may be substituted or unsubstituted, and when substituted, the substituents are preferably one or more groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio.

The term "alkynyl" refers to an alkyl group as defined above consisting of at least two carbon atoms and at least one carbon-carbon triple bond, e.g., ethynyl, propynyl, butynyl, and the like. Alkynyl groups may be substituted or unsubstituted, and when substituted, the substituents are preferably one or more groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio.

The term "cycloalkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent, the cycloalkyl ring containing from 3 to 20 carbon atoms, preferably from 3 to 12 carbon atoms, more preferably from 3 to 6 carbon atoms. Non-limiting examples of monocyclic cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatrienyl, cyclooctyl, and the like; polycyclic cycloalkyl groups include spiro, fused and bridged cycloalkyl groups.

The term "spirocycloalkyl" refers to a 5 to 20 membered polycyclic group sharing one carbon atom (referred to as a spiro atom) between monocyclic rings, which may contain one or more double bonds, but none of the rings have a completely conjugated pi-electron system. Preferably 6 to 14, more preferably 7 to 10. Spirocycloalkyl groups are classified into a single spirocycloalkyl group, a double spirocycloalkyl group or a multi spirocycloalkyl group, preferably a single spirocycloalkyl group and a double spirocycloalkyl group, according to the number of spiro atoms shared between rings. More preferably 4-membered/4-membered, 4-membered/5-membered, 4-membered/6-membered, 5-membered/5-membered or 5-membered/6-membered. Non-limiting examples of spirocycloalkyl groups include:

the term "fused cyclic alkyl" refers to a 5 to 20 membered all carbon polycyclic group in which each ring in the system shares an adjacent pair of carbon atoms with other rings in the system, wherein one or more of the rings may contain one or more double bonds, but none of the rings has a completely conjugated pi-electron system. Preferably 6 to 14, more preferably 7 to 10. They may be classified into bicyclic, tricyclic, tetracyclic or polycyclic fused ring alkyls according to the number of constituent rings, preferably bicyclic or tricyclic, more preferably 5-or 6-membered bicycloalkyl. Non-limiting examples of fused ring alkyl groups include:

the term "bridged cycloalkyl" refers to a 5 to 20 membered all carbon polycyclic group in which any two rings share two carbon atoms not directly attached, which may contain one or more double bonds, but none of the rings have a completely conjugated pi-electron system. Preferably 6 to 14, more preferably 7 to 10. They may be classified as bicyclic, tricyclic, tetracyclic or polycyclic bridged cycloalkyl groups, preferably bicyclic, tricyclic or tetracyclic, more preferably bicyclic or tricyclic, depending on the number of constituent rings. Non-limiting examples of bridged cycloalkyl groups include:

the cycloalkyl ring may be fused to an aryl, heteroaryl or heterocycloalkyl ring, where the ring to which the parent structure is attached is cycloalkyl, non-limiting examples of which include indanyl, tetrahydronaphthyl, benzocycloheptanyl, and the like. Cycloalkyl groups may be optionally substituted or unsubstituted, and when substituted, the substituents are preferably one or more groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, oxo, carboxy or carboxylate.

The term "heterocyclyl" refers to a saturated or partially unsaturated mono-or polycyclic cyclic hydrocarbon substituent containing from 3 to 20 ring atoms wherein one or more of the ring atoms is selected from nitrogen, oxygen, or S (O)_m(wherein m is an integer from 0 to 2) but excludes the ring moiety of-O-O-, -O-S-, or-S-S-, the remaining ring atoms being carbon. Preferably 3 to 12 ring atoms, of which 1 to 4 are heteroatoms; most preferably 3 to 8 ring atoms, of which 1 to 3 are heteroatoms; most preferably 5 to 7 ring atoms, of which 1 to 2 or 1 to 3 are heteroatoms. Non-limiting examples of monocyclic heterocyclyl groups include pyrrolidinyl, imidazolidinyl, tetrahydrofuranyl, tetrahydrothienyl, dihydroimidazolyl, dihydrofuranyl, dihydropyrazolyl, dihydropyrrolyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl, pyranyl, and the like, preferably 1,2, 5-oxadiazolyl, pyranyl, or morpholinyl. Polycyclic heterocyclic groups include spiro, fused and bridged heterocyclic groups.

The term "spiroheterocyclyl" refers to a 5-to 20-membered polycyclic heterocyclic group in which one atom (referred to as the spiro atom) is shared between monocyclic rings, and in which one or more ring atoms is selected from nitrogen, oxygen, or S (O)_m(wherein m is an integer of 0 to 2) and the remaining ring atoms are carbon. It may contain one or more double bonds, but no ring has a completely conjugated pi-electron system. Preferably 6 to 14, more preferably 7 to 10. The spiro heterocyclic group is classified into a mono-spiro heterocyclic group, a di-spiro heterocyclic group or a multi-spiro heterocyclic group, preferably a mono-spiro heterocyclic group and a di-spiro heterocyclic group, according to the number of spiro atoms shared between rings. More preferred are 4-membered/4-membered, 4-membered/5-membered, 4-membered/6-membered, 5-membered/5-membered or 5-membered/6-membered mono spiroheterocyclic groups.Non-limiting examples of spiro heterocyclic groups include:

the term "fused heterocyclyl" refers to a 5 to 20 membered polycyclic heterocyclic group in which each ring in the system shares an adjacent pair of atoms with other rings in the system, one or more rings may contain one or more double bonds, but none of the rings has a fully conjugated pi-electron system in which one or more ring atoms is selected from nitrogen, oxygen or S (O)_m(wherein m is an integer of 0 to 2) and the remaining ring atoms are carbon. Preferably 6 to 14, more preferably 7 to 10. They may be classified into bicyclic, tricyclic, tetracyclic or polycyclic fused heterocyclic groups according to the number of constituent rings, preferably bicyclic or tricyclic, more preferably 5-or 6-membered bicyclic fused heterocyclic groups. Non-limiting examples of fused heterocyclic groups include:

the term "bridged heterocyclyl" refers to a 5 to 14 membered polycyclic heterocyclic group in which any two rings share two atoms not directly attached which may contain one or more double bonds, but none of the rings have a fully conjugated pi-electron system in which one or more of the ring atoms is selected from nitrogen, oxygen or S (O)_m(wherein m is an integer of 0 to 2) and the remaining ring atoms are carbon. Preferably 6 to 14, more preferably 7 to 10. They may be classified into bicyclic, tricyclic, tetracyclic or polycyclic bridged heterocyclic groups according to the number of constituent rings, preferably bicyclic, tricyclic or tetracyclic, more preferably bicyclic or tricyclic. Non-limiting examples of bridged heterocyclic groups include:

the heterocyclyl ring may be fused to an aryl, heteroaryl or cycloalkyl ring, wherein the ring to which the parent structure is attached is heterocyclyl, non-limiting examples of which include:

and the like.

The heterocyclyl group may be optionally substituted or unsubstituted, and when substituted, the substituents are preferably one or more groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, oxo, carboxy or carboxylate.

The term "aryl" refers to a 6 to 14 membered all carbon monocyclic or fused polycyclic (i.e., rings which share adjacent pairs of carbon atoms) group having a conjugated pi-electron system, preferably 6 to 10 membered, such as phenyl and naphthyl. More preferably phenyl. The aryl ring may be fused to a heteroaryl, heterocyclyl or cycloalkyl ring, wherein the ring attached to the parent structure is an aryl ring, non-limiting examples of which include:

the aryl group may be substituted or unsubstituted, and when substituted, the substituent is preferably one or more groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, carboxy or carboxylate.

The term "heteroaryl" refers to a heteroaromatic system comprising 1 to 4 heteroatoms, 5 to 14 ring atoms, wherein the heteroatoms are selected from oxygen, sulfur and nitrogen. Heteroaryl is preferably 5 to 10 membered, containing 1 to 3 heteroatoms; more preferably 5 or 6 membered, containing 1 to 2 heteroatoms; preferably, for example, imidazolyl, furyl, thienyl, thiazolyl, pyrazolyl, oxazolyl, pyrrolyl, tetrazolyl, pyridyl, pyrimidinyl, thiadiazole, pyrazinyl and the like, preferably imidazolyl, thiazolyl, pyrazolyl or pyrimidinyl, thiazolyl; more preferably pyrazolyl or thiazolyl. The heteroaryl ring may be fused to an aryl, heterocyclyl or cycloalkyl ring, wherein the ring joined together with the parent structure is a heteroaryl ring, non-limiting examples of which include:

heteroaryl groups may be optionally substituted or unsubstituted, and when substituted, the substituents are preferably one or more groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, carboxyl or carboxylate groups.

The term "alkoxy" refers to-O- (alkyl) and-O- (unsubstituted cycloalkyl), wherein alkyl is as defined above. Non-limiting examples of alkoxy groups include: methoxy, ethoxy, propoxy, butoxy, cyclopropoxy, cyclobutoxy, cyclopentyloxy, cyclohexyloxy. Alkoxy groups may be optionally substituted or unsubstituted, and when substituted, the substituents are preferably one or more groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, carboxy or carboxylate groups.

The term "haloalkyl" refers to an alkyl group substituted with one or more halogens wherein alkyl is as defined above.

The term "haloalkoxy" refers to an alkoxy group substituted with one or more halogens, wherein the alkoxy group is as defined above.

The term "hydroxyalkyl" refers to an alkyl group substituted with a hydroxy group, wherein alkyl is as defined above.

The term "hydroxy" refers to an-OH group.

The term "halogen" refers to fluorine, chlorine, bromine or iodine.

The term "amino" refers to-NH₂。

The term "cyano" refers to — CN.

The term "nitro" means-NO₂。

The term "oxo" refers to ═ O.

The term "carboxy" refers to-C (O) OH.

The term "mercapto" refers to-SH.

The term "ester group" refers to-C (O) O (alkyl) or-C (O) O (cycloalkyl), wherein alkyl and cycloalkyl are as defined above.

The term "acyl" refers to compounds containing the group-C (O) R, where R is alkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl.

The term "sulfonic acid group" means-S (O)₂OH。

The term "sulfonate group" means-S (O)₂O (alkyl) or-S (O)₂O (cycloalkyl), wherein alkyl and cycloalkyl are as defined above.

"optional" or "optionally" means that the subsequently described event or circumstance may, but need not, occur, and that the description includes instances where the event or circumstance occurs or does not. For example, "a heterocyclic group optionally substituted with an alkyl" means that an alkyl may, but need not, be present, and the description includes the case where the heterocyclic group is substituted with an alkyl and the heterocyclic group is not substituted with an alkyl.

"substituted" means that one or more, preferably up to 5, more preferably 1 to 3, hydrogen atoms in the group are independently substituted with a corresponding number of substituents. It goes without saying that the substituents are only in their possible chemical positions, and that the person skilled in the art is able to determine (experimentally or theoretically) possible or impossible substitutions without undue effort. For example, amino or hydroxyl groups having free hydrogen may be unstable in combination with carbon atoms having unsaturated (e.g., olefinic) bonds.

"pharmaceutical composition" means a mixture containing one or more compounds described herein or a physiologically/pharmaceutically acceptable salt or prodrug thereof in admixture with other chemical components, as well as other components such as physiologically/pharmaceutically acceptable carriers and excipients. The purpose of the pharmaceutical composition is to facilitate administration to an organism, facilitate absorption of the active ingredient and exert biological activity.

"pharmaceutically acceptable salts" refers to salts of the compounds of the present invention which are safe and effective for use in the body of a mammal and which possess the requisite biological activity.

Synthesis of the Compounds of the invention

In order to achieve the purpose of the invention, the invention adopts the following technical scheme.

The preparation method of the compound shown in the general formula I or the salt thereof is as follows.

The compounds of general formula I can be obtained by etherification, acylation and substitution reactions using compound Ia as starting material according to the process of scheme 1.

Synthesis of scheme 1:

reacting compound Ia with Ib at room temperature under an alkaline condition to obtain compound Ic, wherein the alkali reagent is preferably cesium carbonate or potassium carbonate; subsequently, reacting the compound Ic with Z-Cl under low-temperature alkaline conditions to obtain a compound Id, wherein the alkaline reagent is preferably sodium carbonate, and the temperature is preferably-10 ℃; finally, the compound Id is reacted with H-W-Cy-L under high-temperature acidic conditions¹-L²-R⁶The reaction gives the compound of formula I, the acid reagent is preferably trifluoroacetic acid, and the temperature is preferably 100 ℃.

Wherein W, X, Y, Z, Cy, L¹、L²、R¹、R⁶As defined in formula I.

Detailed Description

The present invention is further described below with reference to examples, which are not intended to limit the scope of the present invention.

The structure of the compounds is determined by Nuclear Magnetic Resonance (NMR) or/and Mass Spectrometry (MS). NMR shift at 10^-6The units in (ppm) are given. NMR was measured using a Brukerdps model 300 nuclear magnetic spectrometer using deuterated dimethyl sulfoxide (DMSO-d)₆) DeuteriumChloroform (CDCl)₃) Deuterated methanol (CD)₃OD), internal standard Tetramethylsilane (TMS).

MS was measured using a 1100Series LC/MSD Trap (ESI) mass spectrometer (manufacturer: Agilent).

Liquid phase preparation lc3000 HPLC and lc6000 HPLC (manufacturer: Innovation Consumer) were used.

HPLC was carried out by using Shimadzu LC-20AD high pressure liquid chromatograph (Agilent TC-C18250X 4.6mm 5 μm column) and Shimadzu LC-2010AHT high pressure liquid chromatograph (Phenomenex C18250X 4.6mm 5 μm column).

Average inhibition rate of kinase and IC₅₀The values were measured using a multifunctional staining 3 microplate reader (Biotech, USA).

The thin layer chromatography silica gel plate is Qingdao ocean chemical GF254 silica gel plate, the specification of the silica gel plate used by Thin Layer Chromatography (TLC) is 0.15 mm-0.2 mm, and the specification of the thin layer chromatography separation and purification product is 0.4 mm-0.5 mm.

Column chromatography generally uses Qingdao marine silica gel 100-200 meshes and 200-300 meshes as a carrier.

Known starting materials of the present invention can be synthesized by or according to methods known in the art, or can be purchased from the companies such as cyber-mart, beijing coup, Sigma, carbofuran, yishiming, shanghai kaya, enokay, anigil chemical, and the like.

In the examples, the reaction can be carried out in an argon atmosphere or a nitrogen atmosphere, unless otherwise specified.

An argon atmosphere or nitrogen atmosphere means that the reaction flask is connected to a balloon of argon or nitrogen with a volume of about 1L.

The microwave reaction was carried out using a CEM Discover SP type microwave reactor.

In the examples, the solution means an aqueous solution unless otherwise specified.

In the examples, the reaction temperature is, unless otherwise specified, from 20 ℃ to 30 ℃ at room temperature.

The progress of the reaction in the examples was monitored by Thin Layer Chromatography (TLC) using a developing solvent system of: a: dichloromethane and methanol system, B: n-hexane and ethyl acetate system, C: petroleum ether and ethyl acetate system, D: the volume ratio of acetone and solvent is adjusted according to the polarity of the compound.

The eluent system for column chromatography and the developing agent system for thin-layer chromatography used for purifying compounds comprise: a: dichloromethane and methanol system, B: petroleum ether, ethyl acetate and dichloromethane system, C: the volume ratio of the solvent in the petroleum ether and ethyl acetate system is adjusted according to the different polarities of the compounds, and a small amount of basic or acidic reagents such as triethylamine, acetic acid and the like can be added for adjustment.

Example 1: preparation of 1- (5- ((2- ((4- (4-methylpiperazin-1-yl) phenyl) amino) thieno [3,2-d ] pyrimidin-4-yl) oxy) -3, 4-dihydroquinolin-1 (2H) -yl) prop-2-en-1-one

Step 1: synthesis of 1,2,3, 4-tetrahydroquinolin-5-ol (intermediate 1A)

Quinolin-5-ol (380mg,2.6mmol) was dissolved in ethanol (90mL), and 2N hydrochloric acid (1.27g) was added and stirred at room temperature for 10 minutes. Then, platinum oxide hydrate (90mg,0.4mmol) was added thereto, and hydrogen gas was bubbled through the mixture, and the reaction was stirred at room temperature for 1 hour. After the reaction was complete, suction filtered and the filtrate was concentrated to dryness to give 420mg of the title product as a brown-black solid in yield: 100 percent.

Step 2: synthesis of 2-chloro-4- ((1,2,3, 4-tetrahydroquinolin-5-yl) oxy) thieno [3,2-d ] pyrimidine (intermediate 1B)

2, 4-Dichlorothieno [3,2-d ] pyrimidine (0.58g,2.84mmol), cesium carbonate (2.30g,7.1mmol), and DMSO (4mL) were sequentially added to the reaction flask and stirred at room temperature. Then 1,2,3, 4-tetrahydroquinolin-5-ol (0.528g,2.84mmol) was dissolved in DMSO (6mL) and added dropwise to the reaction flask. After the addition was complete, the reaction was stirred at room temperature overnight. After the reaction was completed, dichloromethane (70mL) and water (70mL) were added for extraction, the organic phase was washed with water (70mL × 2), the organic phase was separated, dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography (eluent: petroleum ether: ethyl acetate: dichloromethane ═ 6:1:10) to obtain 468mg of the title product as a pale yellow solid, yield: 51.9 percent.

And step 3: synthesis of 1- (5- ((2-chlorothieno [3,2-d ] pyrimidin-4-yl) oxy) -3, 4-dihydroquinolin-1 (2H) -yl) prop-2-en-1-one (intermediate 1C)

2-chloro-4- ((1,2,3, 4-tetrahydroquinolin-5-yl) oxy) thieno [3,2-d ] pyrimidine (900mg,2.83mmol), tetrahydrofuran (30mL), distilled water (5mL), and sodium bicarbonate (951mg,11.32mmol) were sequentially added to a reaction flask, and stirred under ice bath. A mixture of acryloyl chloride (308mg,3.40mmol) and tetrahydrofuran (5mL) was slowly added dropwise to the flask, the reaction was carried out in an ice bath for 6 hours, the ice bath was removed, and the reaction was carried out at room temperature overnight. After the reaction was completed, dichloromethane (150mL) was added, the organic phase was washed with 5% saturated brine (100mL × 2), the organic phase was separated, dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography (eluent: petroleum ether: ethyl acetate: dichloromethane ═ 4:1:2-2:1:2) to give 548mg of the title product as a pale yellowish white solid in yield: 52.1 percent.

And 4, step 4: synthesis of 1- (5- ((2- ((4- (4-methylpiperazin-1-yl) phenyl) amino) thieno [3,2-d ] pyrimidin-4-yl) oxy) -3, 4-dihydroquinolin-1 (2H) -yl) prop-2-en-1-one (Compound 1)

1- (5- ((2-chlorothieno [3,2-d ] pyrimidin-4-yl) oxy) -3, 4-dihydroquinolin-1 (2H) -yl) prop-2-en-1-one (260mg,0.7mmol), 2-butanol (6mL), trifluoroacetic acid (80mg,0.7mmol), and 4- (4-methylpiperazin-1-yl) aniline (134mg,0.7mmol) were sequentially added to a reaction flask, and the reaction was stirred at 100 ℃ for 15 hours under argon protection. After the reaction was completed, the temperature was decreased to room temperature, dichloromethane (150mL) was added for dilution, and then washed with 0.5% sodium bicarbonate solution (100mL) and 5% brine (3X 100mL) in this order, the organic layer was separated, dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by preparative liquid phase (eluent: 0% -100% acetonitrile: aqueous solution) to give 88mg of the title product as a pale yellow solid in yield: 23.9 percent.

MS:m/z＝527.1[M+H]⁺.

¹H NMR(300MHz,CDCl₃):ppm 1.90(m,2H),2.42(s,3H),2.67(m,6H),3.15(m,4H),3.86(m,2H),5.74(m,1H),6.45(m,1H),6.51(m,1H),6.62(m,2H),6.76(m,1H),6.94(m,1H),7.09(m,1H),7.28(m,3H),7.83(m,1H).

Example 2: preparation of 1- (5- ((2- ((4-morpholinylphenyl) amino) thieno [3,2-d ] pyrimidin-4-yl) oxy) -3, 4-dihydroquinolin-1 (2H) -yl) prop-2-en-1-one

The title compound was obtained in the same manner as the preparation of example 1 except that 4-morpholinylaniline was used instead of 4- (4-methylpiperazin-1-yl) aniline.

MS:m/z＝514.3[M+H]⁺.

¹H NMR(300MHz,CDCl₃):ppm 2.03(m,2H),2.76(m,2H),3.15(m,4H),3.63(m,4H),3.78(m,2H),5.80(m,1H),6.48(m,1H),6.57(m,1H),6.74(m,2H),6.80(m,1H),6.92(m,1H),7.10(m,1H),7.30(m,3H),7.92(m,1H).

Example 3: preparation of 1- (5- ((2- ((4-thiomorpholinylphenyl) amino) thieno [3,2-d ] pyrimidin-4-yl) oxy) -3, 4-dihydroquinolin-1 (2H) -yl) prop-2-en-1-one

The title compound was obtained in the same manner as the preparation of example 1 except that 4- (thiomorpholin-4-yl) aniline was used instead of 4- (4-methylpiperazin-1-yl) aniline.

MS:m/z＝530.4[M+H]⁺.

¹H NMR(300MHz,CDCl₃):ppm 1.91(m,2H),2.66(m,2H),3.00(m,4H),3.72(m,4H),3.86(m,2H),5.68(m,1H),6.45(m,1H),6.50(m,1H),6.63(m,2H),6.76(m,1H),7.00(m,1H),7.23(m,1H),7.39(m,3H),8.00(m,1H).

Example 4: preparation of 1- (5- ((2- (4- (1, 1-dioxothiomorpholino) phenyl) amino) thieno [3,2-d ] pyrimidin-4-yl) oxy) -3, 4-dihydroquinolin-1 (2H) -yl) prop-2-en-1-one

The title compound was obtained in the same manner as the preparation of example 1 except that 4- (1, 1-thiomorpholine) aniline was used in place of 4- (4-methylpiperazin-1-yl) aniline.

MS:m/z＝562.2[M+H]⁺.

¹H NMR(300MHz,CDCl₃):ppm 1.91(m,2H),2.62(m,2H),3.56(m,4H),3.86(m,2H),4.03(m,4H),5.60(m,1H),6.43(m,1H),6.57(m,1H),6.64(m,2H),6.81(m,1H),7.00(m,1H),7.22(m,1H),7.40(m,3H),7.96(m,1H).

Example 5: preparation of 1- (5- ((2- ((4- (4, 4-difluoropiperidin-1-yl) phenyl) amino) thieno [3,2-d ] pyrimidin-4-yl) oxy) -3, 4-dihydroquinolin-1 (2H) -yl) prop-2-en-1-one

The title compound was obtained in the same manner as the preparation of example 1 except that 4- (4, 4-difluoro-1-piperidinyl) aniline was used instead of 4- (4-methylpiperazin-1-yl) aniline.

MS:m/z＝548.3[M+H]⁺.

¹H NMR(300MHz,CDCl₃):ppm 1.86(m,2H),2.16(m,4H),2.73(m,2H),3.56(m,2H),3.83(m,4H),5.71(m,1H),6.46(m,1H),6.53(m,1H),6.64(m,2H),6.76(m,1H),6.96(m,1H),7.08(m,1H),7.34(m,3H),7.86(m,1H).

Example 6: preparation of 1- (5- ((2- ((4- (piperidin-1-yl) phenyl) amino) thieno [3,2-d ] pyrimidin-4-yl) oxy) -3, 4-dihydroquinolin-1 (2H) -yl) prop-2-en-1-one

The title compound was obtained in the same manner as the preparation of example 1 except that 4- (1-piperidinyl) aniline was used instead of 4- (4-methylpiperazin-1-yl) aniline.

MS:m/z＝512.2[M+H]⁺.

¹H NMR(300MHz,CDCl₃):ppm 1.60(m,2H),1.78(m,4H),1.88(m,2H),2.75(m,2H),3.58(m,2H),3.70(m,4H),5.70(m,1H),6.43(m,1H),6.55(m,1H),6.62(m,2H),6.78(m,1H),6.96(m,1H),7.10(m,1H),7.30(m,3H),7.87(m,1H).

Example 7: preparation of 1- (5- ((2- ((4- (2-methoxyethoxy) phenyl) amino) thieno [3,2-d ] pyrimidin-4-yl) oxy) -3, 4-dihydroquinolin-1 (2H) -yl) prop-2-en-1-one

The title compound was obtained in the same manner as the preparation of example 1 except that 4- (2-methoxyethoxy) aniline was used in place of 4- (4-methylpiperazin-1-yl) aniline.

MS:m/z＝503.2[M+H]⁺.

¹H NMR(300MHz,CDCl₃):ppm 1.80(m,2H),2.70(m,2H),3.18(m,2H),3.40(m,3H),3.74(m,2H),4.33(m,2H),5.74(m,1H),6.29(m,1H),6.48(m,1H),6.64(m,2H),6.82(m,1H),6.94(m,1H),7.15(m,1H),7.42(m,3H),7.79(m,1H).

Example 8: preparation of 1- (5- ((2- (4- (4-methylpiperazin-1-yl) phenyl) amino) thieno [3,2-d ] pyrimidin-4-yl) oxy) -3, 4-dihydroquinolin-1 (2H) -yl) but-2-yn-1-one

Step 1: synthesis of 1- (5- ((2-chlorothieno [3,2-d ] pyrimidin-4-yl) oxy) -3, 4-dihydroquinolin-1 (2H) -yl) but-2-yn-1-one (intermediate 8A)

Preparation of but-2-alkynoyl chloride:

2.0g (23.8mmol) of 2-butynoic acid was added to THF (20mL), and 1.2mL of oxalyl chloride was slowly added dropwise with stirring at room temperature, and after completion of the addition, 3 drops of DMF were added to the reaction mixture, and stirring was continued at room temperature for 1 hour. Concentrating to dryness.

2-chloro-4- ((1,2,3, 4-tetrahydroquinolin-5-yl) oxy) thieno [3,2-d ] pyrimidine (900mg,2.83mmol), tetrahydrofuran (30mL), distilled water (5mL), and sodium bicarbonate (951mg,11.32mmol) were sequentially added to a reaction flask, and stirred under ice bath. A mixture of but-2-ynoyl chloride (349mg,3.40mmol) and tetrahydrofuran (5mL) was slowly added dropwise to the flask, allowed to react for 6 hours in an ice bath, removed from the ice bath, and allowed to react overnight at room temperature. After the reaction was completed, dichloromethane (150mL) was added, the organic phase was washed with 5% saturated brine (100mL × 2), the organic phase was separated, dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography (eluent: petroleum ether: ethyl acetate: dichloromethane ═ 4:1:2-2:1:2) to give 436mg of the title product as a pale yellowish white solid in a yield of 40.1%.

Step 2: synthesis of 1- (5- ((2- (4- (4-methylpiperazin-1-yl) phenyl) amino) thieno [3,2-d ] pyrimidin-4-yl) oxy) -3, 4-dihydroquinolin-1 (2H) -yl) but-2-yn-1-one (Compound 8)

1- (5- ((2-chlorothieno [3,2-d ] pyrimidin-4-yl) oxy) -3, 4-dihydroquinolin-1 (2H) -yl) but-2-yn-1-one (268mg,0.7mmol), 2-butanol (6mL), trifluoroacetic acid (80mg,0.7mmol), and 4- (4-methylpiperazin-1-yl) aniline (134mg,0.7mmol) were successively added to a reaction flask, and the reaction was stirred at 100 ℃ for 15 hours under argon protection. After the reaction was completed, the temperature was reduced to room temperature, dichloromethane (150mL) was added for dilution, and then washed with 0.5% sodium bicarbonate solution (100mL) and 5% brine (3X 100mL) in this order, the organic layer was separated, dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated to dryness under reduced pressure, and the residue was purified by preparative liquid phase (eluent: 0% -100% acetonitrile: aqueous solution) to give 60mg of the title product as a pale yellow solid in yield: 15.9 percent.

MS:m/z＝539.1[M+H]⁺.

¹H NMR(300MHz,CDCl₃):ppm 1.90(m,2H),2.42(s,3H),2.67(m,6H),3.15(m,4H),3.86(m,2H),6.61(m,2H),6.78(m,1H),6.95(m,1H),7.07(m,1H),7.28(m,3H),7.83(m,1H).

Example 9: preparation of 1- (8- ((2- ((4- (4-methylpiperazin-1-yl) phenyl) amino) thieno [3,2-d ] pyrimidin-4-yl) oxy) -2, 3-dihydro-4H-benzo [ b ] [1,4] oxazin-4-yl) but-2-yn-1-one

The title compound was obtained in the same manner as the preparation of example 8 except for using 3, 4-dihydro-2H-benzo [ b ] [1,4] oxazin-8-ol instead of 1,2,3, 4-tetrahydroquinolin-5-ol (intermediate 1A).

MS:m/z＝541.2[M+H]⁺.

¹H NMR(300MHz,CDCl₃):ppm 2.20(s,3H),2.35(s,3H),2.62(m,4H),3.08(m,4H),3.88(m,2H),4.20(m,2H),6.68(m,1H),6.84(m,1H),7.06(m,1H),7.12(m,1H),7.26(m,3H),7.30(m,1H),8.03(m,1H).

Example 10: preparation of 1- (8- ((2- ((4- (4-methylpiperazin-1-yl) phenyl) amino) thieno [3,2-d ] pyrimidin-4-yl) oxy) -2, 3-dihydro-4H-benzo [ b ] [1,4] oxazin-4-yl) prop-2-en-1-one

The title compound was obtained in the same manner as the preparation of example 1 except for using 3, 4-dihydro-2H-benzo [ b ] [1,4] oxazin-8-ol instead of 1,2,3, 4-tetrahydroquinolin-5-ol (intermediate 1A).

MS:m/z＝529.1[M+H]⁺.

¹H NMR(300MHz,CDCl₃):ppm 2.33(s,3H),2.58(m,4H),3.06(m,4H),3.87(m,2H),4.18(m,2H),5.86(m,1H),6.40(m,1H),6.74(m,2H),6.85(m,1H),7.08(m,1H),7.11(m,1H),7.24(m,3H),7.29(m,1H),8.02(m,1H).

Example 11: preparation of 1- (8- ((2- ((4-morpholinylphenyl) amino) thieno [3,2-d ] pyrimidin-4-yl) oxy) -2, 3-dihydro-4H-benzo [ b ] [1,4] oxazin-4-yl) prop-2-en-1-one

The title compound was obtained in the same manner as the preparation of example 2 except for using 3, 4-dihydro-2H-benzo [ b ] [1,4] oxazin-8-ol instead of 1,2,3, 4-tetrahydroquinolin-5-ol (intermediate 1A).

MS:m/z＝516.2[M+H]⁺.

¹H NMR(300MHz,CDCl₃):ppm 3.15(m,4H),3.66(m,4H),3.88(m,2H),4.19(m,2H),5.87(m,1H),6.42(m,1H),6.73(m,2H),6.87(m,1H),7.11(m,1H),7.14(m,1H),7.26(m,3H),7.31(m,1H),8.03(m,1H).

Example 12: preparation of 1- (8- ((2- ((4-thiomorpholinphenyl) amino) thieno [3,2-d ] pyrimidin-4-yl) oxy) -2, 3-dihydro-4H-benzo [ b ] [1,4] oxazin-4-yl) prop-2-en-1-one

The title compound was obtained in the same manner as the preparation of example 3 except for using 3, 4-dihydro-2H-benzo [ b ] [1,4] oxazin-8-ol instead of 1,2,3, 4-tetrahydroquinolin-5-ol (intermediate 1A).

MS:m/z＝532.1[M+H]⁺.

¹H NMR(300MHz,CDCl₃):ppm 2.68(m,4H),3.76(m,4H),3.89(m,2H),4.19(m,2H),5.86(m,1H),6.43(m,1H),6.70(m,2H),6.88(m,1H),7.10(m,1H),7.13(m,1H),7.27(m,3H),7.33(m,1H),8.03(m,1H).

Example 13: preparation of 1- (8- ((2- ((4- (1, 1-thiomorpholinyl) phenyl) amino) thieno [3,2-d ] pyrimidin-4-yl) oxy) -2, 3-dihydro-4H-benzo [ b ] [1,4] oxazin-4-yl) prop-2-en-1-one

The title compound was obtained in the same manner as the preparation of example 4 except for using 3, 4-dihydro-2H-benzo [ b ] [1,4] oxazin-8-ol instead of 1,2,3, 4-tetrahydroquinolin-5-ol (intermediate 1A).

MS:m/z＝564.1[M+H]⁺.

¹H NMR(300MHz,CDCl₃):ppm 3.38(m,4H),3.66(m,4H),3.88(m,2H),4.20(m,2H),5.83(m,1H),6.42(m,1H),6.71(m,2H),6.86(m,1H),7.08(m,1H),7.12(m,1H),7.25(m,3H),7.32(m,1H),8.05(m,1H).

Example 14: preparation of 1- (8- ((2- ((4- (4, 4-difluoropiperidin-1-yl) phenyl) amino) thieno [3,2-d ] pyrimidin-4-yl) oxy) -2, 3-dihydro-4H-benzo [ b ] [1,4] oxazin-4-yl) prop-2-en-1-one

The title compound was obtained in the same manner as the preparation of example 5 except for using 3, 4-dihydro-2H-benzo [ b ] [1,4] oxazin-8-ol instead of 1,2,3, 4-tetrahydroquinolin-5-ol (intermediate 1A).

MS:m/z＝550.2[M+H]⁺.

¹H NMR(300MHz,CDCl₃):ppm 2.02(m,4H),3.10(m,4H),3.78(m,2H),4.07(m,2H),5.90(m,1H),6.42(m,1H),6.78(m,2H),6.87(m,1H),7.08(m,1H),7.12(m,1H),7.25(m,3H),7.43(m,1H),8.00(m,1H).

Example 15: preparation of 1- (8- ((2- ((4- (piperidin-1-yl) phenyl) amino) thieno [3,2-d ] pyrimidin-4-yl) oxy) -2, 3-dihydro-4H-benzo [ b ] [1,4] oxazin-4-yl) prop-2-en-1-one

The title compound was obtained in the same manner as the preparation of example 6 except for using 3, 4-dihydro-2H-benzo [ b ] [1,4] oxazin-8-ol instead of 1,2,3, 4-tetrahydroquinolin-5-ol (intermediate 1A).

MS:m/z＝514.2[M+H]⁺.

¹H NMR(300MHz,CDCl₃):ppm 1.83(m,2H),2.02(m,4H),3.16(m,4H),3.68(m,2H),4.05(m,2H),5.73(m,1H),6.34(m,1H),6.70(m,2H),6.83(m,1H),7.05(m,1H),7.13(m,1H),7.22(m,3H),7.29(m,1H),8.03(m,1H).

Example 16: preparation of 1- (5- ((2- (4- (4-methylpiperazin-1-yl) phenoxy) thieno [3,2-d ] pyrimidin-4-yl) oxy) -3, 4-dihydroquinolin-1 (2H) -yl) prop-2-en-1-one

Step 1: synthesis of 1- (5- ((2- (4- (4-methylpiperazin-1-yl) phenoxy) thieno [3,2-d ] pyrimidin-4-yl) oxy) -3, 4-dihydroquinolin-1 (2H) -yl) prop-2-en-1-one (Compound 16)

1- (5- ((2-chlorothieno [3,2-d ] pyrimidin-4-yl) oxy) -3, 4-dihydroquinolin-1 (2H) -yl) prop-2-en-1-one (520mg,1.4mmol), DMF (6mL), potassium carbonate (387mg,2.8mmol) and (4-methylpiperazin-1-yl) phenol (323mg,1.7mmol) were successively added to a reaction flask, and the reaction was stirred at 100 ℃ overnight. After the reaction was completed, the temperature was reduced to room temperature, ethyl acetate (150mL) was added for dilution, and then washed with water (3X 100mL) in order, the organic layer was separated, dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by preparative liquid phase (eluent: 0% -100% acetonitrile: aqueous solution) to give 236mg of the title product as a pale yellow solid in yield: 31.9 percent.

MS:m/z＝528.1[M+H]⁺.

¹H NMR(300MHz,CDCl₃):ppm 1.82(m,2H),2.36(s,3H),2.64(m,6H),3.13(m,4H),3.76(m,2H),5.68(m,1H),6.35(m,1H),6.48(m,1H),6.68(m,2H),6.74(m,1H),6.84(m,1H),7.19(m,1H),7.28(m,3H),8.01(m,1H).

Example 17: preparation of 1- (5- ((2- (4-morpholinylphenoxy) thieno [3,2-d ] pyrimidin-4-yl) oxy) -3, 4-dihydroquinolin-1 (2H) -yl) prop-2-en-1-one

The title compound was obtained in the same manner as the preparation of example 16 except that 4-morpholinophenol was used instead of (4-methylpiperazin-1-yl) phenol.

MS:m/z＝515.3[M+H]⁺.

¹H NMR(300MHz,CDCl₃):ppm 2.13(m,2H),2.86(m,2H),3.15(m,4H),3.58(m,4H),3.82(m,2H),5.70(m,1H),6.38(m,1H),6.57(m,1H),6.68(m,2H),6.80(m,1H),6.92(m,1H),7.12(m,1H),7.33(m,3H),7.85(m,1H).

Example 18: preparation of 1- (5- ((2- (4- (piperidin-1-yl) phenoxy) thieno [3,2-d ] pyrimidin-4-yl) oxy) -3, 4-dihydroquinolin-1 (2H) -yl) prop-2-en-1-one

The title compound was obtained in the same manner as the preparation of example 16 except that 4- (piperidin-1-yl) phenol was used instead of (4-methylpiperazin-1-yl) phenol.

MS:m/z＝513.2[M+H]⁺.

¹H NMR(300MHz,CDCl₃)：ppm 1.62(m,2H),1.74(m,4H),1.92(m,2H),2.75(m,2H),3.58(m,2H),3.76(m,4H),5.66(m,1H),6.42(m,1H),6.57(m,1H),6.64(m,2H),6.78(m,1H),7.00(m,1H),7.11(m,1H),7.30(m,3H),7.98(m,1H).

Example 19: preparation of 1- (5- ((2- (4- ((4-methylpiperazin-1-yl) methyl) phenoxy) thieno [3,2-d ] pyrimidin-4-yl) oxy) -3, 4-dihydroquinolin-1 (2H) -yl) prop-2-en-1-one

The title compound was obtained in the same manner as the preparation of example 16 except for using 4- (4-methylpiperazin-1-ylmethyl) phenol instead of (4-methylpiperazin-1-yl) phenol.

MS:m/z＝542.2[M+H]⁺.

¹H NMR(300MHz,CDCl₃)：ppm 1.66(m,2H),1.82(m,2H),2.25(m,3H),2.34(m,4H),2.55(m,4H),3.18(m,2H),3.76(m,2H),5.67(m,1H),6.44(m,1H),6.58(m,1H),6.71(m,2H),6.79(m,1H),6.96(m,1H),7.14(m,1H),7.36(m,3H),8.06(m,1H).

Example 20: preparation of 1- (5- ((2- (4- (4-methylpiperazine-1-carbonyl) phenoxy) thieno [3,2-d ] pyrimidin-4-yl) oxy) -3, 4-dihydroquinolin-1 (2H) -yl) prop-2-en-1-one

The title compound was obtained in the same manner as the preparation of example 16 except for using 1- (4-hydroxybenzoyl) -4-methylpiperazine instead of (4-methylpiperazin-1-yl) phenol.

MS:m/z＝556.2[M+H]⁺.

¹H NMR(300MHz,CDCl₃)：ppm 1.62(m,2H),1.92(m,2H),2.19(m,3H),2.35(m,4H),3.48(m,2H),3.56(m,4H),5.58(m,1H),6.44(m,1H),6.58(m,1H),6.68(m,2H),6.78(m,1H),7.06(m,1H),7.41(m,1H),7.70(m,3H),7.98(m,1H).

Example 21: preparation of 1- (8- ((2- (4- (4-methylpiperazin-1-yl) phenoxy) thieno [3,2-d ] pyrimidin-4-yl) oxy) -2, 3-dihydro-4H-benzo [ b ] [1,4] oxazin-4-yl) prop-2-en-1-one

The title compound was obtained in the same manner as the preparation of example 16 except for using 3, 4-dihydro-2H-benzo [ b ] [1,4] oxazin-8-ol instead of 1,2,3, 4-tetrahydroquinolin-5-ol (intermediate 1A).

MS:m/z＝530.1[M+H]⁺.

¹H NMR(300MHz,CDCl₃):ppm 2.43(s,3H),2.62(m,4H),3.14(m,4H),3.79(m,2H),4.20(m,2H),5.76(m,1H),6.52(m,1H),6.76(m,2H),6.89(m,1H),7.03(m,1H),7.09(m,1H),7.34(m,3H),7.39(m,1H),8.08(m,1H).

Example 22: preparation of 1- (8- ((2- (4-morpholinylphenoxy) thieno [3,2-d ] pyrimidin-4-yl) oxy) -2, 3-dihydro-4H-benzo [ b ] [1,4] oxazin-4-yl) prop-2-en-1-one

The title compound was obtained in the same manner as the preparation of example 17 except for using 3, 4-dihydro-2H-benzo [ b ] [1,4] oxazin-8-ol instead of 1,2,3, 4-tetrahydroquinolin-5-ol (intermediate 1A).

MS:m/z＝517.2[M+H]⁺.

¹H NMR(300MHz,CDCl₃):ppm 3.08(m,4H),3.59(m,4H),3.86(m,2H),4.24(m,2H),5.86(m,1H),6.52(m,1H),6.75(m,2H),6.90(m,1H),7.11(m,1H),7.15(m,1H),7.27(m,3H),7.30(m,1H),8.13(m,1H).

Example 23: preparation of 1- (8- ((2- (4- (piperidin-1-yl) phenoxy) thieno [3,2-d ] pyrimidin-4-yl) oxy) -2, 3-dihydro-4H-benzo [ b ] [1,4] oxazin-4-yl) prop-2-en-1-one

The title compound was obtained in the same manner as the preparation of example 18 except for using 3, 4-dihydro-2H-benzo [ b ] [1,4] oxazin-8-ol instead of 1,2,3, 4-tetrahydroquinolin-5-ol (intermediate 1A).

MS:m/z＝515.2[M+H]⁺.

¹H NMR(300MHz,CDCl₃):ppm 1.88(m,2H),2.34(m,4H),3.26(m,4H),3.67(m,2H),4.15(m,2H),5.78(m,1H),6.40(m,1H),6.68(m,2H),6.86(m,1H),7.05(m,1H),7.14(m,1H),7.23(m,3H),7.36(m,1H),8.06(m,1H).

Example 24: preparation of 1- (5- ((2- ((4- (4-methylpiperazin-1-yl) phenyl) amino) thieno [3,2-d ] pyrimidin-4-yl) amino) -3, 4-dihydroquinolin-1 (2H) -yl) prop-2-en-1-one

Step 1: synthesis of 2-chloro-N- (1,2,3, 4-tetrahydroquinolin-5-yl) thieno [3,2-d ] pyrimidin-4-amine (intermediate 24A)

2, 4-dichlorothieno [3,2-d ] pyrimidine (2g,9.81mmol), 2-butanol (20mL), trifluoroacetic acid (1.12g,9.81mmol), 1,2,3, 4-tetrahydroquinolin-5-amine (1.45g,9.81mmol) were added to a reaction flask in this order and stirred at 100 ℃ for 15 hours under argon. After the reaction was completed, the temperature was decreased to room temperature, dichloromethane (150mL) was added for dilution, and then washed with 0.5% sodium bicarbonate solution (100mL) and 5% brine (3X 100mL) in this order, the organic layer was separated, dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by preparative liquid phase (eluent: 0% -100% acetonitrile: aqueous solution) to give 0.5g of the title product as a pale yellow solid in yield: 16.1 percent.

Step 2: synthesis of 1- (5- ((2-chlorothieno [3,2-d ] pyrimidin-4-yl) amino) -3, 4-dihydroquinolin-1 (2H) -yl) prop-2-en-1-one (intermediate 24B)

2-chloro-N- (1,2,3, 4-tetrahydroquinolin-5-yl) thieno [3,2-d ] pyrimidin-4-amine (500mg,1.58mmol), tetrahydrofuran (10mL), distilled water (2mL), and sodium bicarbonate (265mg,3.16mmol) were sequentially added to a reaction flask, and stirred under ice bath. A mixture of acryloyl chloride (172mg,1.90mmol) and tetrahydrofuran (5mL) was slowly added dropwise to the flask, the reaction was carried out in an ice bath for 6 hours, the ice bath was removed, and the reaction was carried out at room temperature overnight. After the reaction was completed, dichloromethane (150mL) was added, the organic phase was washed with 5% saturated brine (100mL × 2), the organic phase was separated, dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography (eluent: petroleum ether: ethyl acetate ═ 3:1) to give 260mg of the title product as a pale yellow solid, yield: 44.4 percent.

And step 3: synthesis of 1- (5- ((2- ((4- (4-methylpiperazin-1-yl) phenyl) amino) thieno [3,2-d ] pyrimidin-4-yl) amino) -3, 4-dihydroquinolin-1 (2H) -yl) prop-2-en-1-one (Compound 24)

1- (5- ((2-chlorothieno [3,2-d ] pyrimidin-4-yl) amino) -3, 4-dihydroquinolin-1 (2H) -yl) prop-2-en-1-one (260mg,0.7mmol), 2-butanol (6mL), trifluoroacetic acid (80mg,0.7mmol), and 4- (4-methylpiperazin-1-yl) aniline (134mg,0.7mmol) were sequentially added to a reaction flask, and the reaction was stirred at 100 ℃ for 15 hours under argon protection. After the reaction was completed, the temperature was reduced to room temperature, dichloromethane (150mL) was added for dilution, and then washed successively with 0.5% sodium bicarbonate solution (100mL) and 5% brine (3X 100mL), to give an organic layer, which was dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by the preparative liquid phase (eluent: 0% -100% acetonitrile: aqueous solution) to give 46mg of the title product as a pale yellow solid in yield: 12.5 percent.

MS:m/z＝526.1[M+H]⁺.

¹H NMR(300MHz,CDCl₃)：ppm 2.10(m,2H),2.38(s,3H),2.75(m,6H),3.26(m,4H),3.88(m,2H),5.76(m,1H),6.43(m,1H),6.54(m,1H),6.67(m,2H),6.78(m,1H),6.94(m,1H),7.11(m,1H),7.31(m,3H),7.86(m,1H).

Biological evaluation of Compounds of the invention

Test example 1: evaluation of BTK inhibitory Activity

To a 384 reaction plate (6008280, Perkin Elmer) was added 2. mu.L/well of BTK solution (available from Promega). Test compounds (10mM stock) were diluted 100-fold to 100 μ M with 100% DMSO, in 384 dilution plates (3657, corning) at 1: 3, and the gradient concentration of the test compound is 100, 33.3, 11.1, 3.7, 1.24, 0.41, 0.14, 0.046, 0.015, 0.0051 and 0 μ M. mu.L of test compound was transferred to 384 dilution plates containing 38. mu.L of 1 Xkinase reaction buffer, mixed well and centrifuged at 1000 rpm. mu.L of the test compound was transferred to 384 reaction plates, centrifuged at 1000rpm and incubated at 25 ℃ for 15 minutes. mu.L of the substrate mixture (ATP: 10mM, 4. mu.L; Poly E4Y 1: 1mg/mL, 20. mu.L; kinase reaction buffer: 776. mu.L) was transferred to a 384 reaction plate, centrifuged at 1000rpm and incubated for 60 minutes at 25 ℃. The final concentration gradient of the test compound in the reaction system is 1000, 333.3, 111.1, 37.03, 12.35, 4.14, 1.37, 0.46, 0.15, 0.051 and 0 nM. The final concentration of DMSO is 1%. mu.L of ADP-Glo solution (from Promega) was transferred to 384 reaction plates, centrifuged at 1000rpm and incubated at 25 ℃ for 40 minutes. Transfer 10. mu.L of Detection solution to 384 reaction plates, centrifuge at 1000rpm and incubate at 25 ℃ for 40 min. Rlu (relative luminescence determination) signals were read using an Envision multifunctional plate reader (Perkin Elmer), and signal intensity was used to characterize the extent of BTK kinase activity. IC by Using Microsoft Excel₅₀Calculation and analysis of (3). The test results are shown in table 1.

In Table 1, A means IC₅₀<100 nM; b is IC₅₀100nM to 500 nM; c means IC₅₀500nM to 1,000 nM; d means IC₅₀>1,000nM。

TABLE 1 BTK inhibitory Activity of the Compounds of the invention

And (4) conclusion: as shown in the above table, the compounds of the present invention all showed excellent BTK inhibitory activity.

Test example 2: effect of Compounds of the invention on type II collagen-induced DBA/1J mouse arthritis model (CIA)

The CIA model is an animal model widely used for studying the activity of drugs for treating rheumatoid arthritis in humans.

Animals: DBA/1 mice, sex: male, number: 30, body weight: 14-16g, week age range: 6-7 weeks old, purchased from Witonglihua laboratory animal technology, Inc., Beijing, SPF grade, animal production license number: SCXK (Jing) 2012-0001, issuing unit: the scientific and technical committee of Beijing.

Grouping: model control group, administration (30mg/Kg) group (Compound of example 10)

An appropriate amount of the Bovine Type II Collage (from Chondrex. Inc.) was taken, dissolved in 0.05M acetic acid (4mg collagen/mL) and refrigerated overnight at 4 ℃. The mixture was thoroughly emulsified with an equal amount of complete Freund's adjuvant in an ice bath environment, and each DBA/1J mouse was injected subcutaneously 1.5cm from the root of the tail with 0.1mL (200. mu.g of collagen) of the emulsion. Clinical symptom observation and arthritis scoring were performed starting at 4 weeks after molding, scores of all animals showing symptoms were ranked and randomly grouped layer by layer, and divided into 2 groups, namely a model control group (oral gavage given with an equivalent amount of vehicle 0.5% CMC) and an administration group (oral gavage given with 30mg/kg of the compound of example 10), 1 administration/day, and continued administration for 21 days, and arthritis scoring and weight change were measured every 3 days after administration.

Arthritis index score: the arthritis index was scored according to Wood's arthritis scoring criteria, as follows.

0 minute: is normal

1 minute: red swelling involving 1 finger joint

And 2, dividing: the mild red swelling of more than 2 finger joints or the entire paw

And 3, dividing: severe red and swollen foot and paw

And 4, dividing: severe redness and swelling of the paw, stiffness of the joints, and lack of elasticity.

Lesions were divided between 0 and 4 for each of the 4 paws and the total integral of the limb was calculated. The integrals (arthritis index) at different times were compared.

TABLE 2 Effect of the Compounds of example 10 of the invention on the arthritis score in CIA model animals

Note that: p <0.05, P <0.01, compared to model control group

And (4) conclusion: the compound provided by the invention can be used for remarkably reducing the arthritis score of a CIA model animal, and the compound is proved to have a remarkable improvement effect on the arthritis symptom of a CIA mouse.

Claims

1. A compound shown in a general formula I or a pharmaceutically acceptable salt thereof,

wherein:

x is selected from O or NH;

R¹selected from hydrogen;

y is selected from CH₂Or O;

z is

R³And R⁴Each independently selected from hydrogen;

w is selected from NH;

cy is selected from C₆-C₁₀An aryl group;

L¹and L²Each independently selected from one or more of single bond or-O-;

R⁶is selected from C₁-C₆Alkyl or 5 to 7 heterocyclyl; wherein said C₁-C₆Alkyl is optionally further substituted by C₁-C₆Alkoxy substitution; said 5 to 7 heterocyclyl is optionally further substituted by one or more groups selected from halogen, C₁-C₆Alkyl groups.

2. The compound of formula I or a pharmaceutically acceptable salt thereof according to claim 1, which is a compound of formula II or a pharmaceutically acceptable salt thereof,

wherein R is¹、R⁶、Y、Z、Cy、L¹、L²As defined in claim 1.

3. The compound of formula I or a pharmaceutically acceptable salt thereof according to claim 1, which is a compound of formula IV or a pharmaceutically acceptable salt thereof,

wherein R is¹、R⁶、Y、Z、Cy、L¹、L²As defined in claim 1.

4. A compound of general formula I or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 3, wherein Cy is phenyl.

5. A compound of general formula I or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 3, wherein R⁶Selected from piperazinyl, morpholinyl, piperidinyl, thiomorpholinyl, optionally further substituted by one or more groups selected from halo, C₁-C₆Alkyl groups.

6. A compound, or a pharmaceutically acceptable salt thereof, wherein the compound is selected from:

1- (5- ((2- ((4- (4-methylpiperazin-1-yl) phenyl) amino) thieno [3,2-d ] pyrimidin-4-yl) oxy) -3, 4-dihydroquinolin-1 (2H) -yl) prop-2-en-1-one;

1- (5- ((2- ((4-morpholinylphenyl) amino) thieno [3,2-d ] pyrimidin-4-yl) oxy) -3, 4-dihydroquinolin-1 (2H) -yl) prop-2-en-1-one;

1- (5- ((2- ((4-thiomorpholinylphenyl) amino) thieno [3,2-d ] pyrimidin-4-yl) oxy) -3, 4-dihydroquinolin-1 (2H) -yl) prop-2-en-1-one;

1- (5- ((2- ((4- (4, 4-difluoropiperidin-1-yl) phenyl) amino) thieno [3,2-d ] pyrimidin-4-yl) oxy) -3, 4-dihydroquinolin-1 (2H) -yl) prop-2-en-1-one;

1- (5- ((2- ((4- (2-methoxyethoxy) phenyl) amino) thieno [3,2-d ] pyrimidin-4-yl) oxy) -3, 4-dihydroquinolin-1 (2H) -yl) prop-2-en-1-one;

1- (8- ((2- ((4- (4-methylpiperazin-1-yl) phenyl) amino) thieno [3,2-d ] pyrimidin-4-yl) oxy) -2, 3-dihydro-4H-benzo [ b ] [1,4] oxazin-4-yl) prop-2-en-1-one;

1- (8- ((2- ((4-morpholinylphenyl) amino) thieno [3,2-d ] pyrimidin-4-yl) oxy) -2, 3-dihydro-4H-benzo [ b ] [1,4] oxazin-4-yl) prop-2-en-1-one;

1- (8- ((2- ((4- (piperidin-1-yl) phenyl) amino) thieno [3,2-d ] pyrimidin-4-yl) oxy) -2, 3-dihydro-4H-benzo [ b ] [1,4] oxazin-4-yl) prop-2-en-1-one;

1- (5- ((2- ((4- (4-methylpiperazin-1-yl) phenyl) amino) thieno [3,2-d ] pyrimidin-4-yl) amino) -3, 4-dihydroquinolin-1 (2H) -yl) prop-2-en-1-one.

7. A process for the preparation of a compound of formula I according to claim 1 or a pharmaceutically acceptable salt thereof, comprising the steps of:

reacting the compound Id with a compound H-W-Cy-L under high-temperature acidic conditions¹-L²-R⁶Reacting to obtain a compound shown in a general formula I;

wherein W, X, Y, Z, Cy, L¹、L²、R¹、R⁶As defined in claim 1.

8. The method of claim 7, wherein the agent that provides acidity is trifluoroacetic acid.

9. The process according to claim 7 or 8, wherein the reaction temperature is 100 ℃.

10. A pharmaceutical composition comprising a compound of general formula I according to any one of claims 1 to 5 or a pharmaceutically acceptable salt thereof or a compound of claim 6 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

11. Use of a compound of general formula I according to any one of claims 1 to 5 or a pharmaceutically acceptable salt thereof or a compound according to claim 6 or a pharmaceutically acceptable salt thereof or a pharmaceutical composition according to claim 10 for the preparation of a BTK kinase inhibitor.

12. Use of a compound of general formula I according to any one of claims 1 to 5 or a pharmaceutically acceptable salt thereof or a compound according to claim 6 or a pharmaceutically acceptable salt thereof or a pharmaceutical composition according to claim 10 for the preparation of a medicament for the prevention and/or treatment of a disease associated with BTK kinase activity.

13. The use of claim 12, wherein the disease is selected from inflammation, autoimmune disease, or cancer.

14. The use of claim 13, wherein the inflammation is arthritis, inflammatory enteritis, or uveitis.

15. The use of claim 14, wherein the arthritis is rheumatoid arthritis or psoriatic arthritis.

16. The use of claim 13, wherein the autoimmune disease is multiple sclerosis, lupus, psoriasis, or sarcoidosis.

17. The use of claim 13, wherein the cancer is a solid tumor.

18. The use of claim 13, wherein the cancer is breast cancer, cervical cancer, colon cancer, lung cancer, gastric cancer, rectal cancer, pancreatic cancer, brain cancer, skin cancer, oral cancer, prostate cancer, bone cancer, kidney cancer, ovarian cancer, bladder cancer, liver cancer, fallopian tube tumor, ovarian tumor, peritoneal tumor, melanoma, glioma, glioblastoma, hepatocellular carcinoma, papillary renal tumor, head and neck tumor, leukemia, lymphoma, myeloma, or non-small cell lung cancer.