CN109776544B - Pyrazolo [3,4-d ] pyrimidine compound and preparation method and application thereof - Google Patents

Abstract

The compound is an inhibitor of Bruton's tyrosine kinase. In addition, the invention describes pharmaceutical compositions and formulations comprising the compounds, and the use of such kinase inhibitors, alone or in combination with other compounds, for the treatment of kinase-mediated or kinase-dependent conditions.

Description

Pyrazolo [3,4-d ] pyrimidine compound and preparation method and application thereof

Technical Field

The present invention relates to compounds, methods for preparing compounds, pharmaceutical compositions and medicaments of compounds, and the use of compounds for the treatment, prevention, diagnosis of Bruton's tyrosine kinase (Btk) related diseases, disorders or conditions.

Background

Protein tyrosine kinase regulates a series of physiological and biochemical processes of cell growth, differentiation, apoptosis and the like by controlling a signal transduction pathway of the cell. Abnormal kinase activity has been implicated in a number of human diseases including inflammatory, autoimmune and cancer diseases. Abnormal expression of protein tyrosine kinase has been found in common human cancers (such as gastric cancer, lung cancer, lymphoma, etc.), and protein tyrosine kinase has become one of the important targets for research and development of antitumor drugs.

Btk is a member of the Tec family of non-receptor tyrosine kinases and consists of a PH domain, a TH domain, an SH3 domain, an SH2 domain, and a catalytic domain 5 moiety. Btk participates in various signal pathways, plays an important role in regulating and controlling proliferation, differentiation and apoptosis of cells, is activated depending on Syk and Lyn along with activation of BCR in the signal pathway connected with a cell surface B Cell Receptor (BCR), can cause activation of downstream signals including MAPK, NF kB and the like, and abnormal BCR-mediated signal transduction can cause mis-regulated B cell activation or formation of pathogenic autoantibodies, which can cause various autoimmune or inflammatory diseases. Continued activation of Btk is a prerequisite for the development of Chronic Lymphocytic Leukemia (CLL), and its aberrant expression also promotes survival of activated B-cell subsets in diffuse large B-cell lymphoma (DLBCL).

The Btk small molecule inhibitor can inhibit the proliferation of B lymphoma cells and promote the apoptosis of tumor cells by inhibiting the activity of Btk, can also inhibit the generation of B cell autoantibodies and cytokines, and has good prospects for treating hematological malignancies and autoimmune dysregulated diseases. A number of compounds have been introduced into clinical studies for the treatment of B-cell lymphomas, leukemias, and the like. Ibrutinib (Ibrutinib) developed by Pharmacyclics biopharmaceutical company is used as the first irreversible Btk small-molecule inhibitor drug on the market, and has obvious curative effects for treating mantle cell lymphoma, chronic lymphocytic leukemia, macroglobulinemia and the like at present. Other small molecule Btk inhibitors in clinical development are ONO-4059 from ONO, ACP-196 from Acerta, and CC-292 from Celgene, among others.

At present, the excellent clinical results of ibrutinib show that the high-selectivity small-molecule inhibitor of Btk kinase will become another hot spot in the field of global new drug development, and the pyrazolo [3,4-d ] pyrimidine structural compound is an orally effective Btk small-molecule inhibitor drug. Therefore, in view of the urgent need of treating tumor diseases, the research on the pyrazolo [3,4-d ] pyrimidine compounds with diversified structures has important significance for developing new Btk small-molecule inhibitor drugs with better effects.

Disclosure of Invention

The present invention relates to compounds, pharmaceutical compositions, medicaments and methods that are useful (a) for the diagnosis, prevention, treatment, or prevention of diseases, disorders, or symptoms associated with Btk; (b) alleviating a side effect or symptom associated with Btk; (c) controlling a disease, disorder or symptom associated with Btk. In one aspect, the methods, compounds, pharmaceutical compositions, and medicaments set forth herein consist of inhibitors that inhibit Btk.

In a first aspect of the invention, there is provided a compound of formula I, or a pharmaceutically acceptable salt, solvate, active metabolite, polymorph, ester, isomer or prodrug thereof:

wherein:

L₁is selected from unsubstituted or substituted C1-C6 alkyl, unsubstituted or substituted C2-C6 alkenyl and unsubstituted or substituted C2-C6 alkynyl;

L₂is selected from- (CH)₂)_n-、-(CH₂)_n-O-、-(CH₂)_n-S-、-(CH₂)_n-NH-, wherein n is 0, 1,2,3 or 4;

ar is selected from unsubstituted or substituted aryl or heteroaryl;

y is selected from unsubstituted or substituted alkyl, or a 4-, 5-, 6-membered cycloalkyl ring;

R₁selected from H or lower alkyl;

or, Y and N and R₁Are connected to form a four-element, five-element or six-element heterocyclic ring;

g is selected from the group consisting of H,

wherein R is₂、R₃And R₄Each independently selected from H, halogen, -COOH, unsubstituted or substituted lower alkyl, unsubstituted or substituted lower heteroalkyl.

In another preferred embodiment, wherein: n is 1 or 2.

In another preferred embodiment, wherein: ar is selected from unsubstituted or substituted aryl; more preferably, the substitution is meta or para; more preferably, the substituent is alkoxy or halogen.

In another preferred embodiment, wherein: l is₁Is selected from unsubstituted C2-C6 alkenyl or unsubstituted C2-C6 alkynyl; more preferably, wherein: l is₁Selected from ethenyl or ethynyl.

In another preferred embodiment, wherein: y and N are linked to R1 to form a six membered heterocyclic ring.

In another preferred embodiment, wherein: g is

In another preferred embodiment, the present invention provides a compound selected from the group consisting of:

in another preferred embodiment, the compounds provided by the present invention have inhibitory activity against Bruton's tyrosine kinase (Btk).

In a second aspect of the invention, there is provided a pharmaceutical composition comprising a compound provided by the invention as described above and a pharmaceutically acceptable excipient.

In another preferred embodiment, the above pharmaceutical composition is in the form of an aqueous dispersion, liquid, gel, syrup, elixir, syrup, suspension, aerosol, controlled-release agent, quick-dissolving agent, effervescent agent, lyophilized agent, tablet, powder, pill, coated tablet, capsule, delayed-release agent, extended-release agent, pulsatile controlled-release agent, multiparticulate agent, or immediate-release agent.

In a third aspect of the present invention, there is provided a process for the preparation of a compound provided by the present invention as described above, said process comprising the steps of: under the protection of nitrogen, mixing a compound shown as a structure in a formula I-d with DMF (dimethyl formamide), a palladium catalyst and organic base, further reacting with a compound shown as a structure in a formula I-c at 30-150 ℃ for 3-15 hours to obtain a compound shown as a structure in a formula I-e, deprotecting the compound under an acidic condition to prepare an intermediate shown as a structure in a formula I-f, and then carrying out amide condensation reaction with G-Z in the presence of alkali or a condensing agent and an alkane solvent to obtain the compound provided by the invention;

wherein: l is₁、L₂、Ar、Y、R₁G is defined as in claim 1; z is selected from halogen or hydroxyl.

In a fourth aspect of the invention, there is provided the use of a compound provided by the invention as described above in the preparation of a medicament for the prevention or treatment of inflammation, autoimmune diseases associated with abnormal B cell proliferation (such as rheumatoid arthritis) and/or neoplastic diseases.

Therefore, the invention provides a novel Btk small molecule inhibitor drug with better effect.

Drawings

Fig. 1 shows that a homogeneous phase time-resolved fluorescence (HTRF) method establishes the principle of kinase activity detection of small molecule inhibitors of Btk.

Fig. 2 shows that a homogeneous phase time-resolved fluorescence (HTRF) method establishes a kinase activity detection procedure for small molecule inhibitors of Btk.

Detailed Description

Term(s) for

Unless otherwise defined, terms used in this application, including the specification and claims, are defined as follows. It must be noted that, in the specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. Conventional methods of mass spectrometry, nuclear magnetism, HPLC, protein chemistry, biochemistry, recombinant DNA technology and pharmacology are used, if not otherwise stated. In this application, "or" and "means" and/or "are used unless otherwise stated.

"Compound of formula (I)" means a compound of formula (I).

"alkyl" refers to an aliphatic hydrocarbon group. The alkyl moiety may be saturated (meaning not containing any unsaturated units such as carbon-carbon double bonds or carbon-carbon triple bonds) or the alkyl moiety may be unsaturated (meaning containing at least one unsaturated unit). The alkyl moiety, whether saturated or unsaturated, may be branched or straight chain.

An "alkyl" moiety (moity) may have from 1 to 8 carbon atoms (as long as appearing herein, a numerical range such as "1 to 8" refers to each integer in the given range, e.g., "1 to 8 carbon atoms" refers to alkyl groups that may contain 1 carbon atom, 2 carbon atoms, 3 carbon atoms, etc. up to 8 carbon atoms, although the present definition also encompasses the term in the absence of a given numerical rangeThe occurrence of "alkyl"). The alkyl group of the compounds described herein may be designated as "C₁-C₆Alkyl "or the like. By way of example "C₁-C₆Alkyl "means one, two, three, four, five or six carbon atoms in the alkyl chain. Typical alkyl groups include, but are not limited to, methyl, ethyl, propylisopropyl, butylisobutyl, tert-butyl, pentyl, hexyl and the like. The term "lower alkyl" is similarly used for groups having 1 to 4 carbon atoms.

An "alkoxy" group refers to an "alkyl" O-group, alkyl being as defined herein.

"amido" is a chemical moiety of the formula-C (═ O) NHR or-NHC (═ O) R, selected from the group consisting of alkyl, cycloalkyl, aryl, heteroaryl (attached through a ring carbon) and heteroalicyclic hydrocarbon (attached through a ring carbon). The amide may be an amino acid or a peptide molecule linked to a compound of formula (I) to form a prodrug. Any amino or carbon side chain in the compounds described herein is optionally amidated as desired. See Greene and Wuts, Protective Groups in Organic Synthesis, 3^rd Ed.，John Wiley&Sons，New York，NY，1999。

The term "aryl" as used herein refers to an aromatic ring wherein each atom forming the ring is a carbon atom. The ring of the aryl group is composed of five, six, seven, eight, nine or more atoms. The aryl group is optionally substituted. In one aspect, aryl is phenyl or naphthyl. Depending on the structure, the aryl group may be a mono-radical or a di-radical (e.g., arylene). In one aspect, aryl is C₆-C₁₀And (4) an aryl group.

The term "cycloalkyl" refers to a monocyclic or polycyclic aliphatic hydrocarbon, a non-aromatic radical, in which each atom (e.g., a backbone atom) forming the ring is a carbon atom. Cycloalkyl groups may be saturated or partially unsaturated. The cycloalkyl group may be attached to the aromatic ring at a point on a carbon atom other than that of the aromatic ring. Cycloalkyl groups comprise from 3 to 10 ring-forming atoms. In certain particular aspects, the cycloalkyl group is selected from cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. Cycloalkyl groups may be substituted or unsubstituted.

The term "ester" refers to a chemical moiety bearing a-COOR group, wherein R is selected from the group consisting of alkyl, cycloalkyl, aryl, heteroaryl (attached through a ring carbon) and heteroalicyclic hydrocarbon (attached through a ring carbon). Any of the hydroxy or carboxy side chains in the compounds described herein are esterified, if desired. Examples of procedures and specific Groups for preparing such esters are described in Greene and Wuts, Protective Groups in Organic Synthesis, 3^rd Ed.，John Wiley&Sons，New York，NY，1999。

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

The term "heteroalkyl" refers to an alkyl group in which one or more of the backbone atoms is selected from an atom other than carbon, such as oxygen, nitrogen, sulfur, phosphorus, or both. In one aspect, heteroalkyl is C₁-C₆A heteroalkyl group.

The term "heterocycle" or "heterocyclic" refers to heteroaromatic rings (also known as heteroaryl groups) and heterocycloalkyl groups (also known as heteroalicyclic groups) containing one to four heteroatoms each selected from O, S and N, each heterocyclyl group containing 4 to 10 atoms in the ring system, but no ring can contain two adjacent O or S atoms. Non-aromatic heterocyclic groups (also referred to as heterocycloalkyl) include groups having only 3 atoms in the ring, but arylheterocyclic groups must have at least 5 atoms in the ring. Heterocyclyl includes benzo ring systems. Examples of ternary heterocyclic groups are aziridinyl; examples of quaternary heterocyclic groups are azetidinyl; examples of five-membered heterocyclic groups are thiazolyl; examples of six-membered heterocyclic groups are pyridyl; an example of a ten-membered heterocyclic group is quinolinyl. Examples of non-aromatic heterocyclic groups are pyrrolidinyl, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothienyl, oxazolidinoneyl, tetrahydropyranyl, dihydropyranyl, tetrahydrothiopyranyl, piperidinyl, morpholinyl, thiomorpholinyl, thioalkyl, piperazinyl, aziridinyl, azetidinyl, oxetanyl, thietanyl, piperidyl, oxepanyl, thietanyl, diazepinyl, thiazepinyl, 1,2,3, 6-tetrahydropyridinyl, pyrrolin-2-yl, pyrrolin-3-yl, indolinyl, 2H-pyranyl, 4H-pyranyl, dioxanyl, 1, 3-dioxolanyl, pyrazolinyl, dithianyl, dithiopentyl, dihydropyranyl, dihydrothienyl, dihydrofuranyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, 3-azabicyclo [3.1.0] hexanyl, 3-azabicyclo [4.1.0] heptanyl, 3H-indolyl and quinolizinyl. Examples of aryl heterocyclic groups are pyridyl, imidazolyl, pyrimidinyl, pyrazolyl, triazolyl, pyrazinyl, tetrazolyl, furyl, thienyl, isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, pyrrolyl, quinolyl, isoquinolyl, indolyl, benzimidazolyl, benzofuryl, cinnoline, indazolyl, indolizinyl, naphthyridinyl, pyridazinyl, triazinyl, isoindolyl, pteridinyl, purinyl, oxadiazolyl, thiadiazoyl, furazanyl, benzofurazanyl, benzothienyl, benzothiazolyl, benzoxazolyl, quinazolinyl, quinoxalinyl, naphthyridinyl, furylpyridinyl (furylpyridinyl). The foregoing groups may be carbon-attached or nitrogen-attached. For example, a group derived from pyrrole may be pyrrol-1-yl (link N) or pyrrol-3-yl (link C). Further, the groups derived from imidazole may be imidazol-1-yl or imidazol-3-yl (both attached to N) or imidazol-2-yl, imidazol-4-yl or imidazol-5-yl (both attached to C). Heterocyclyl includes benzo-fused ring systems. Non-aromatic heterocycles may be substituted by one or two oxygen (═ O) moieties, such as pyrrolin-2-one.

The term "heteroaryl" or "heteroaromatic" refers to a compound that includes one or more ring heteroatoms selected from nitrogen, oxygen, and sulfur. More preferably, heteroaryl groups include indole, azaindole, pyrrole, pyrazole, pyrimidine, pyrazine, pyridine, quinoline, thiophene and furan. In one aspect, a heteroaryl group contains 0 to 3 nitrogen atoms. In another aspect, a heteroaryl group contains 0 to 3 nitrogen atoms, 0 to 1 oxygen atoms, and 0 to 1 sulfur atoms. In another aspect, heteroaryl is monocyclic or bicyclic heteroaryl.

"Heterocycloalkyl", "heteroalicyclic" or "non-aromatic heterocycle" means a cycloalkyl group containing at least one heteroatom selected from nitrogen, oxygen and sulfur. The free radical may be fused to an aryl or heteroaryl group. In certain embodiments, the heterocycloalkyl group is selected from the group consisting of oxazolidinyl, pyrrolidinyl, tetrahydrofuranyl, tetrahydrothienyl, tetrahydropyranyl, tetrahydropalmitanyl, and mixtures thereofThiopyranyl, piperidinyl, morpholinyl, thiomorpholinyl, piperazinyl and indolinyl. The term heteroalicyclic also includes all sugar rings including, but not limited to, monosaccharides, disaccharides, and oligosaccharides. In one aspect, heterocycloalkyl is C₂-C₁₀A heterocycloalkyl group. In another aspect, heterocycloalkyl is C₄-C₁₀A heterocycloalkyl group. In one aspect, the heterocycloalkyl group contains from 0 to 2 nitrogen atoms. In another aspect, the heterocycloalkyl group contains 0 to 2 nitrogen atoms, 0 to 2 oxygen atoms, or 0 to 1 sulfur atom.

The term "bond" refers to a chemical bond between two atoms or between two moieties when the atoms connected by the bond are considered part of a larger structure. In one aspect, when a group described herein is a bond, the absence of a reference group allows for the formation of a bond between the remaining defined groups.

The term "membered ring" includes any cyclic structure. The term "element" is intended to mean the number of backbone atoms constituting a ring. Thus, for example, cyclohexyl, pyridyl, pyranyl, thiopyranyl are six-membered rings, cyclopentyl, pyrrolyl, furanyl and thienyl are five-membered rings.

The term "optionally substituted" or "substituted" means that the reference group may be substituted with one or more additional groups individually and independently selected from the group consisting of alkyl, cycloalkyl, aryl, heteroaryl, heteroalicyclic hydrocarbon, hydroxy, alkoxy, alkylthio, arylthio, alkylsulfinyl, arylsulfonyl, alkylsulfonyl, arylsulfonyl, cyano, halo, carbonyl, thiocarbonyl, nitro, haloalkyl, fluoroalkyl and amino, including mono-and di-substituted amino groups and protected derivatives thereof. By way of illustration, the optional substitution may be halide, -CN, -NO₂Or L_sR_sWherein each L_sIndependently selected from a bond, -O-, -C (O) O-, -S-, -S (O)₂-，-NH-，-NHC(＝O)-，-C(＝O)NH-，S(＝O)₂NH-，-NHS(＝O)₂-OC (- ═ O) NH-, -NHC (- ═ O) O-, or- (C)₁-C₆Alkyl groups); each R_sSelected from hydrogen, alkyl, fluoroalkyl, heteroalkyl, cycloalkyl, aryl, heteroaryl, or heterocycloalkyl. Can be formed as aboveProtecting groups for protected derivatives of substituents reference may be made to Greene and Wuts. In one aspect, the optional substituents are selected from halogen, CF₃，OH，CN，NO₂，SO₃H，SO₂NH₂，SO₂Me，NH₂，COOH，CONH₂Alkoxy, -N (CH)₃)₂And an alkyl group.

In certain embodiments, the compounds have one or more stereocenters, and each center independently exists in R or S form. Reference herein to compounds includes all diastereomeric, enantiomeric, epimeric and appropriate mixtures thereof. Stereoisomers can be obtained by methods such as separation of stereoisomers by chiral chromatography columns.

The methods and formulae described herein include the use of N-oxides (if appropriate), crystalline forms (also known as polymorphs) or pharmaceutically acceptable salts of compounds having the structure of formula (I), and active metabolites of these compounds having the same activity. In some cases, the compounds may exist as tautomers. All tautomers are included within the scope of the compounds mentioned herein. In a particular embodiment, the compounds are present in solvated forms, pharmaceutically acceptable solvents such as water, ethanol and the like. In other embodiments, the compounds are present in unsolvated forms.

Specific pharmaceutical and medical terms

The term "acceptable", as used herein, means that a prescribed component or active ingredient does not unduly adversely affect the health of the general therapeutic target.

The term "Btk dependent," as used herein, refers to a condition that may not cause, or not cause, the same degree of disorder or symptom in the absence of Btk.

The term "Btk-mediated," as used herein, refers to a disease or condition that results in the presence of Btk, and the like may occur in the absence of Btk.

The term "cancer", as used herein, refers to an uncontrolled abnormal growth of cells and under certain conditions is capable of metastasizing (spreading). This type of cancer includes, but is not limited to, solid tumors (e.g., bladder, intestine, brain, chest, uterus, heart, kidney, lung, lymphoid tissue (lymphoma), ovary, pancreas or other endocrine organs (e.g., thyroid), prostate, skin (melanoma), or hematologic tumors (e.g., non-leukemias).

The term "co-administration" or similar terms, as used herein, refers to the administration of several selected therapeutic agents to a patient, either in the same or different modes of administration, at the same or different times.

The term "enhance" or "capable of enhancing", as used herein, means that the desired result can be increased or prolonged, both in potency and duration. Thus, in enhancing the therapeutic effect of a drug, the term "capable of enhancing" refers to the ability of the drug to increase or prolong the potency or duration of the drug in the system. As used herein, "potentiating value" refers to the ability to maximize the enhancement of another therapeutic agent in an ideal system.

The term "inflammatory disease" refers to a condition characterized by one or more of the following conditions. Such as pain, fever, redness, swelling, temporary or permanent loss of function. Inflammation has many manifestations, including, but not limited to, acute, viscous, atrophic, catarrhal, chronic, sclerosing, diffuse, disseminated, exudative, fibrogenic, fibroblastic, topical, granulomatous, proliferative, hypertrophic, interstitial, metastatic, necrotic, occlusive, substantive, plastic, productive, proliferative, pseudomembranous, purulent, sclerosing, serofibrinous, plasmatic, simple, specific, subacute, purulent, toxic, traumatic, and/or ulcerative. Further, inflammatory diseases include, but are not limited to, the following vascular manifestations (polyarteritis, temporal arteritis); joint manifestations (arthritis-crystallinity, skeletal, psoriasis, reactive, rheumatic, reliant); gastrointestinal manifestations (colitis); skin (dermatitis); or the appearance of various tissues and organs (systemic lupus erythematosus).

The term "immunological disorder" refers to a disease or condition that produces an adverse or deleterious response to an endogenous or exogenous antigen. The result is often a dysfunction of the cells, or destruction thereof and dysfunction, or destruction of organs or tissues that may produce immune symptoms.

As used herein, a "metabolite" of a compound refers to a derivative produced during the metabolism of the compound. The term "active metabolite" refers to a biologically active derivative produced during the metabolism of a compound. The term "metabolism," as used herein, refers to the entire process by which a particular substance is converted by an organism (including, but not limited to, hydrolysis reactions and reactions catalyzed by enzymes). Thus, enzymes can specifically alter the structure of a compound. For example, cytochrome P540 catalyzes a series of oxidation and reduction reactions, while uridine diphosphate glucuronyl transferase catalyzes the attachment of an activated glucuronic acid molecule to aryl alcohols, aliphatic alcohols, carboxylic acids, amines and free sulfhydryl groups. Metabolite as used herein refers to the administration of a compound to a host, followed by analysis of a tissue sample from the host; or the compound and liver cells are incubated in vitro, and then the reacted compound is analyzed.

The term "subject" or "patient" includes mammals and non-mammals. Mammals include, but are not limited to, mammals: human, non-human primates such as orangutans, apes, and monkeys; agricultural animals such as cattle, horses, goats, sheep, pigs; domestic animals such as rabbits, dogs; the experimental animals include rodents, such as rats, mice, guinea pigs and the like. Non-mammalian animals include, but are not limited to, birds, fish, and the like. In a preferred embodiment, the mammal of choice is a human.

The terms "treat," "treatment process," or "therapy" as used herein include alleviating, inhibiting, or ameliorating a symptom or condition of a disease; inhibiting the generation of complications; ameliorating or preventing underlying metabolic syndrome; inhibiting the development of a disease or condition, such as controlling the development of a disease or condition; alleviating the disease or symptoms; regression of the disease or symptoms; alleviating a complication caused by the disease or symptom, or preventing or treating a symptom caused by the disease or symptom.

As used herein, a compound or pharmaceutical composition, when administered, can ameliorate a disease, symptom, or condition, particularly severity, delay onset, slow progression, or reduce duration of a condition. Whether fixed or temporary, sustained or intermittent, may be due to or associated with administration.

Compound (I)

In one aspect of the invention, there is provided a compound of formula I, or a pharmaceutically acceptable salt, solvate, active metabolite, polymorph, ester, isomer or prodrug thereof:

wherein L is₁Is selected from unsubstituted or substituted C1-C6 alkyl, unsubstituted or substituted C2-C6 alkenyl and unsubstituted or substituted C2-C6 alkynyl;

L₂is selected from- (CH)₂)_n-、-(CH₂)_n-O-、-(CH₂)_n-S-、-(CH₂)_n-NH-, wherein n is 0, 1,2,3 or 4;

ar is selected from unsubstituted or substituted aryl, unsubstituted or substituted heteroaryl;

the unsubstituted or substituted alkyl group of C1-C6 represents an unsubstituted or substituted straight or branched chain saturated hydrocarbon group including straight or branched chain groups of 1 to 6 carbon atoms, including but not limited to methyl, ethyl, n-propyl, isopropyl, cyclopropyl, n-butyl, cyclobutyl, isobutyl, tert-butyl, n-pentyl, isopentyl, or cyclopentyl; the unsubstituted or substituted alkenyl group having C2-C6 represents an unsubstituted or substituted straight or branched chain alkenyl group including straight or branched chain groups having 2 to 6 carbon atoms, including but not limited to cis-or trans-ethylene, propylene, 1-butene, 2-butene, 1, 3-butadiene, 1-pentene, 2-pentene, 1, 3-pentadiene, 1, 4-pentadiene, 1-hexene, 2-hexene, 3-hexene, 1, 3-hexadiene, 1, 4-hexadiene, 1, 5-hexadiene, 2, 4-hexadiene, 1,3, 5-hexatriene; the unsubstituted or substituted alkynyl group of C2-C6 represents a straight chain or branched alkynyl group having 1 or more triple bonds, including, but not limited to, acetylene, propyne, 1-butyne, 2-butyne, 1, 3-butyne, 1-pentyne, 2-pentyne, 1, 3-pentadiyne, 1, 4-pentadiyne, 1-hexyne, 2-hexyne, 3-hexyne, 1, 4-hexyne, 1, 5-hexyne, 2, 4-hexyne, 1,3, 5-hexyne.

Wherein, the above-mentioned alkyl, alkenyl, alkynyl, aryl, heteroaryl and- (CH)₂)_n-optionally substituted with one or more of the following various substituent groups: hydroxy, halogen, -C_1-4Alkyl and-OC_1-4An alkyl group; the halogen is selected from fluorine, chlorine, bromine or iodine.

In certain embodiments, L₁Is selected from unsubstituted or substituted cis-or trans-C2-C6 alkenyl and unsubstituted or substituted C2-C6 alkynyl.

In certain embodiments, L₁Selected from unsubstituted or substituted cis-or trans-C2 alkenyl and unsubstituted or substituted C2 alkynyl, L₂Is selected from- (CH)₂)_n-、-(CH₂)_n-O-、-(CH₂)_n-S-、-(CH₂)_n-NH-, wherein n is 0, 1,2,3 or 4; further preferably n is 1 or 2.

In certain embodiments, L₁Selected from unsubstituted or substituted cis-or trans-C2 alkenyl and unsubstituted or substituted C2 alkynyl, L₂Is selected from- (CH)₂)_n-、-(CH₂)_n-O-、-(CH₂)_n-S-、-(CH₂)_n-NH-, wherein n is 1 or 2. Ar is selected from unsubstituted or substituted aryl and unsubstituted or substituted heteroaryl.

In certain embodiments, L₁Selected from unsubstituted or substituted cis-or trans-C2 alkenyl and unsubstituted or substituted C2 alkynyl, L₂Is selected from- (CH)₂)_n-、-(CH₂)_n-O-、-(CH₂)_n-S-、-(CH₂)_n-NH-, wherein n is 1 or 2. Ar is selected from unsubstituted or substituted aryl.

Pyrazolo [3,4-d ] as shown in formula I]Pyrimidines wherein Y is selected from unsubstituted or substituted alkyl, or a 4-, 5-, 6-membered cycloalkyl ring; and R is₁Selected from H or lower alkyl;

or, Y and R₁May be joined together to form a 4-, 5-or 6-membered heterocyclic ring;

g is selected from the group consisting of H,

wherein R is₂、R₃And R₄Each independently selected from H, halogen, -COOH, unsubstituted or substituted lower alkyl, unsubstituted or substituted lower heteroalkyl;

in some embodiments of the present invention, the first and second electrodes are,

is selected from

In still further embodiments of the present invention, in some embodiments,

is selected from

In still yet more particular embodiments, the first and second portions of the substrate,

is selected from

In certain embodiments, G is

In some casesIn a specific embodiment, G is

Wherein R is₂Selected from the group consisting of H, -COOH and lower alkyl optionally substituted with: halogen, -OH, -O-lower alkyl, amino, monoalkylamino, dialkylamino, heterocycloalkylamino, alkanoyloxy, alkylsulfonylamino;

in certain embodiments, G is

Wherein R is₂Selected from H and lower alkyl.

In other embodiments, G is selected from

Any combination of groups for the different variables described above is contemplated herein.

Compounds of formula (I) include, but are not limited to, the descriptions in table 1.

Table 1.

Synthesis of Compounds

The compounds of formula (I) described above may be synthesized using standard synthetic techniques or known techniques in combination with the methods described herein. In addition, the solvents, temperatures and other reaction conditions mentioned herein may vary.

The starting materials for the synthesis of the compounds of formula (I) may be synthesized or obtained from commercial sources, such as, but not limited to, Aldrich Chemical co. (Milwaukee, Wis.) or Sigma Chemical co. (st. The compounds described herein and other related compounds having various substituents can be synthesized using well-known techniques and starting materials, including those found in March, ADVANCED ORGANIC CHEMISTRY 4^thEd., (Wiley 1992); carey and Sundberg, ADVANCED ORGANIC CHEMISTRY 4^thEd, Vols.A and B (Plenum 2000, 2001), Green and Wuts, PROTECTIVE GROUPS IN ORGANIC synthieSIS 3^rdThe method in ed., (Wiley 1999). The general method of compound preparation may be varied by the use of appropriate reagents and conditions for introducing different groups into the formulae provided herein.

The compounds of formula (I) described herein are synthesized in a synthetic route as shown in the following scheme, and in some embodiments, the compounds described herein can be prepared by the methods described below. The following methods and examples are intended to illustrate these methods. These schemes and examples should not be construed as limiting the invention in any way. The compounds described herein can also be synthesized using standard synthetic techniques known to those skilled in the art, or using methods known in the art in combination with those described herein.

The synthesis method of the compound I comprises the following steps:

step A: i-c Synthesis: carrying out substitution reaction on a compound I-a and a compound I-b in a solvent under the action of inorganic base to obtain a compound I-c;

wherein, in the structure of the compound I-a, L₁Selected from unsubstituted or substitutedThe C1-C6 alkyl group, the unsubstituted or substituted C2-C6 alkenyl group and the unsubstituted or substituted C2-C6 alkynyl group; - (CH)₂)_nN in-is 0, 1,2,3 or 4; x is selected from halogen; the structure of compound I-b wherein Ar is selected from unsubstituted or substituted aryl and unsubstituted or substituted heteroaryl; r₂Selected from hydroxyl, amino or mercapto; compounds I-c in the structure L₂Is selected from- (CH)₂)_n-、-(CH₂)_n-O-、-(CH₂)_n-S-、-(CH₂)_n-NH-, wherein n is 0, 1,2,3 or 4; preferably, L₁Selected from unsubstituted or substituted cis or trans C2 alkenyl and unsubstituted or substituted C2 alkynyl; - (CH)₂)_nN in-is 0, 1 or 2; x is selected from bromine; ar is selected from unsubstituted or substituted aryl; r₂Selected from hydroxyl; l is₂Is selected from- (CH)₂)_n-O-, wherein n is 0, 1 or 2.

The process for preparing compounds I-c may be conventional in the art for such substitution reactions, and the following reaction methods and conditions are particularly preferred in the present invention: mixing the compound I-b with an organic solvent and an inorganic base, and further carrying out a substitution reaction with the compound I-a to obtain a compound I-c;

in the method for preparing the compound I-c, the solvent is preferably one or more of an ether solvent, an alcohol solvent, a nitrile solvent and acetone, DMF (dimethyl formamide) and water; acetone and DMF are further preferred. The ether solvent is preferably tetrahydrofuran. The alcohol solvent is preferably methanol and/or ethanol. The nitrile solvent is preferably acetonitrile.

In the process for preparing the compound I-c, the volume-to-mass ratio of the solvent to the compound I-b is preferably 5mL/g to 50mL/g, and more preferably 10mL/g to 30 mL/g.

In the process for preparing compound I-c, the inorganic base is preferably one or more of potassium hydroxide, potassium carbonate, sodium hydroxide, sodium carbonate and cesium carbonate, and further preferably potassium carbonate.

In the process for preparing the compound I-c, the molar ratio of the inorganic base to the compound I-b is preferably 1:1 to 5:1, more preferably 1:1 to 2: 1.

In the process for preparing the compound I-c, the molar ratio of the compound I-a to the compound I-b is preferably 1:1 to 5:1, more preferably 1:1 to 2: 1.

In the process for preparing the compounds I-c, the temperature of the substitution reaction is preferably from 0 ℃ to 150 ℃, further preferably from 30 ℃ to 70 ℃.

In the process for preparing the compounds I-c, the progress of the substitution reaction can be monitored by a conventional test method in the art (e.g., TLC or HPLC), and generally the end point of the reaction is determined as the disappearance of the compounds I-b, and the reaction time is preferably 0.5h to 16h, more preferably 3h to 10 h.

Preference is given to including any of the following work-up steps in the process for preparing compounds I-c: when the solvent is selected from DMF, the method (1) is preferably employed. When the solvent is selected from acetone, the method (2) is preferably employed. The method (1) comprises the following steps: after the reaction is finished, adding an organic solvent and water, washing an organic phase by using water and saturated sodium chloride, drying, concentrating and carrying out column chromatography. The method (2) comprises the following steps: after the reaction is finished, filtering, concentrating and carrying out column chromatography. Among them, the organic solvent is more preferably ethyl acetate. The method and conditions of column chromatography are selected according to the methods and conditions of column chromatography which are conventional in the art.

And B: i-e Synthesis: and (3) carrying out a coupling reaction of the compound I-c and the compound I-d in the presence of a transition metal catalyst and a base in a solvent which does not have negative influence on the reaction under the protection of nitrogen to prepare the compound I-e.

Wherein, the structures of the compound I-c, the compound I-d and the compound I-e are L₁Is selected from unsubstituted or substituted C1-C6 alkyl, unsubstituted or substituted C2-C6 alkenyl and unsubstituted or substituted C2-C6 alkynyl; l is₂Is selected from- (CH)₂)_n-、-(CH₂)_n-O-、-(CH₂)_n-S-、-(CH₂)_n-NH-, wherein n is 0, 1,2,3 or 4(ii) a Ar is selected from unsubstituted or substituted aryl and unsubstituted or substituted heteroaryl; y is selected from unsubstituted or substituted alkyl, or a 4-, 5-, 6-membered cycloalkyl ring; and R is₁Selected from H or lower alkyl; or, Y and R₁May be joined together to form a 4-, 5-or 6-membered heterocyclic ring; preferably, L₁Selected from unsubstituted or substituted cis or trans C2 alkenyl and unsubstituted or substituted C2 alkynyl; l is₂Is selected from- (CH)₂)_n-O-, wherein n is 0, 1 or 2; ar is selected from unsubstituted or substituted aryl;

is selected from

More preferably still, the first and second liquid crystal compositions are,

is selected from

The process for preparing compounds I-e may be conventional in the art for such coupling reactions, and the following reaction processes and conditions are particularly preferred in the present invention: under the protection of nitrogen, mixing the compound I-d with an organic solvent, a transition metal catalyst and alkali, and further reacting with the compound I-c at 30-150 ℃ (preferably 80 ℃) for 3-15 hours (preferably 10 hours) to perform coupling reaction to obtain a compound I-e;

in the process for preparing the compounds I-e, any solvent which does not adversely affect the reaction may be used as the solvent. Preferably includes one or more of hydrocarbon solvents (e.g., benzene, toluene and xylene), nitrile solvents, ether solvents (e.g., dimethoxyethane, tetrahydrofuran and 1, 4-dioxane), alcohol solvents, aprotic polar solvents (e.g., DMF, DMSO and hexamethylphosphoramide) and water; DMF is more preferable. The ether solvent is preferably 1, 4-dioxane. The alcohol solvent is preferably methanol and/or ethanol. The nitrile solvent is preferably acetonitrile.

In the process for producing the compound I-e, the volume-to-mass ratio of the solvent to the compound I-d is preferably 5mL/g to 50mL/g, and more preferably 10mL/g to 30 mL/g.

In the process for preparing compounds I-e, examples of the base include: organic bases such as triethylamine, diisopropylethylamine, pyridine, lutidine, collidine, 4-dimethylaminopyridine, potassium tert-butyrate, sodium methoxide, sodium ethoxide, lithium hexamethyldisilazide, sodium hexamethyldisilazide, potassium hexamethyldisilazide and butyllithium; and inorganic bases such as sodium hydrogen carbonate, sodium carbonate, potassium carbonate, cesium carbonate, sodium hydroxide and sodium hydride. Further preferred are triethylamine and diisopropylethylamine.

In the process for preparing the compounds I-e, the molar ratio of the base to the compound I-d is preferably 1:1 to 5:1, more preferably 1:1 to 2: 1.

In the process for preparing the compound I-e, the molar ratio of the compound I-c to the compound I-d is preferably 1:1 to 5:1, and more preferably 3:1 to 5: 1.

In the method for preparing the compounds I-e, examples of transition metal catalysts that can be used in this step include palladium catalysts (e.g., palladium acetate, tris (dibenzylideneacetone) dicesium, and 1, 1' -bis (diphenylphosphino) ferrocene-palladium (II) dichloride-dichloromethane complex, etc.). If desired, ligands (e.g., triphenylphosphine and tri-tert-butylphosphine) may be added, and copper reagents (e.g., cuprous iodide and copper acetate) may be used as co-catalysts. Further preferably, the transition metal catalyst is tetrakis (triphenylphosphine) palladium and the cocatalyst is cuprous iodide.

In the process for producing the compounds I-e, the molar ratio of the compounds I-d to the transition metal catalyst is preferably 0.0001 to 1, and more preferably 0.01 to 0.5. The molar ratio of the compound I-d to the ligand is preferably 0.0001 to 4, more preferably 0.01 to 0.2. The molar ratio of the compound I-d to the cocatalyst is preferably from 0.0001 to 4, more preferably from 0.01 to 0.2.

In the process for preparing the compounds I to e, the temperature of the coupling reaction is preferably 0 ℃ to the temperature of solvent reflux, and more preferably room temperature to 150 ℃.

In the process for preparing the compounds I-e, the progress of the coupling reaction can be monitored by a conventional test method in the art (e.g., TLC or HPLC), and generally the end point of the reaction is determined as the disappearance of the compounds I-d, and the reaction time is preferably 0.5h to 20h, more preferably 3h to 15 h.

In the process for preparing the compounds I-e, the subsequent steps may be carried out after or without isolation and purification by known means of sharing and purification such as concentration, vacuum concentration, crystallization, solvent extraction, precipitation and column chromatography.

The process for preparing compounds I-e preferably comprises the following work-up steps: and after the reaction is finished, adding water and an organic solvent, washing an organic phase by using water and saturated sodium chloride, drying the organic phase by using anhydrous sodium sulfate, carrying out suction filtration, concentrating, and purifying by using column chromatography. Among them, the organic solvent is more preferably ethyl acetate or dichloromethane. The method and conditions of column chromatography are selected according to the methods and conditions of column chromatography which are conventional in the art.

And C: i-f Synthesis: in a solvent, under the action of acid, carrying out deprotection reaction on the compound I-e to obtain a compound I-f;

wherein, the structures of the compounds I-e and I-f are L₁Is selected from unsubstituted or substituted C1-C6 alkyl, unsubstituted or substituted C2-C6 alkenyl and unsubstituted or substituted C2-C6 alkynyl; l is₂Is selected from- (CH)₂)_n-、-(CH₂)_n-O-、-(CH₂)_n-S-、-(CH₂)_n-NH-, wherein n is 0, 1,2,3 or 4; ar is selected from unsubstituted or substituted aryl and unsubstituted or substituted heteroaryl;y is selected from unsubstituted or substituted alkyl, or a 4-, 5-, 6-membered cycloalkyl ring; and R is₁Selected from H or lower alkyl; or, Y and R₁May be joined together to form a 4-, 5-or 6-membered heterocyclic ring; preferably, L₁Selected from unsubstituted or substituted cis or trans C2 alkenyl and unsubstituted or substituted C2 alkynyl; l is₂Is selected from- (CH)₂)_n-O-, wherein n is 0, 1 or 2; ar is selected from unsubstituted or substituted aryl;

is selected from

More preferably still, the first and second liquid crystal compositions are,

is selected from

The method for preparing compounds I-f may be a conventional method for such deprotection in the art, and the following reaction methods and conditions are particularly preferred in the present invention: mixing the compound I-e with an organic solvent and an acid, and performing deprotection to obtain a compound I-f;

in the method for preparing the compounds I-f, the solvent is preferably one or more of an ether solvent, an alcohol solvent, a nitrile solvent and acetone, DMF, ethyl acetate, dichloromethane and water; further preferred are ethyl acetate and dichloromethane. The ether solvent is preferably tetrahydrofuran. The alcohol solvent is preferably methanol and/or ethanol. The nitrile solvent is preferably acetonitrile.

In the process for producing the compounds I-f, the volume-to-mass ratio of the solvent to the compounds I-e is preferably 5mL/g to 50mL/g, and more preferably 10mL/g to 30 mL/g.

In the process for preparing compounds I-f, the acid is preferably an organic or inorganic acid selected from formic acid, acetic acid, trifluoroacetic acid, malic acid, maleic acid, fumaric acid, succinic acid, benzoic acid, methanesulfonic acid, benzenesulfonic acid, acetic acid, propionic acid, butyric acid, caprylic acid, adipic acid, oxalic acid, malonic acid, succinic acid, tartaric acid, phenylacetic acid, phthalic acid, terephthalic acid, valeric acid, caproic acid, capric acid, stearic acid, palmitic acid, acrylic acid, and the like; the inorganic acid is hydrochloric acid, hydrobromic acid, phosphoric acid, hydrogen phosphate, sulfuric acid or hydrogen sulfate and the like. Further, hydrochloric acid or trifluoroacetic acid is preferable.

In the process for preparing the compounds I-f, the molar ratio of the acid to the compound I-e is preferably 1:1 to 20:1, more preferably 5:1 to 10: 1.

In the process for producing the compounds I to f, the temperature of the deprotection reaction is preferably 0 ℃ to 150 ℃, and further preferably room temperature.

In the process for preparing the compounds I-f, the progress of the deprotection reaction can be monitored by a conventional test method in the art (such as TLC or HPLC), and generally the end point of the reaction is set as the disappearance of the compounds I-e, and the reaction time is preferably 0.5h to 20h, more preferably 3h to 12 h.

The process for preparing the compounds I-f preferably comprises the following work-up steps: and after the reaction is finished, adding an organic solvent, pulping, filtering by suction, and drying. Among them, the organic solvent is preferably an alcohol solvent, an ether solvent, a hydrocarbon solvent, or the like. Ethanol and petroleum ether are more preferred.

Step D: i synthesis: in a solvent, under the action of alkali and/or a condensing agent, carrying out amide forming reaction on a compound I-f and G-Z to obtain a compound I;

wherein, the structures of the compounds I-f and the compound I are L₁Is selected from unsubstituted or substituted C1-C6 alkyl, unsubstituted or substituted C2-C6 alkenyl and unsubstituted or substituted C2-C6 alkynyl; l is₂Is selected from- (CH)₂)_n-、-(CH₂)_n-O-、-(CH₂)_n-S-、-(CH₂)_n-NH-, wherein n is 0, 1,2,3 or 4; ar is selected from unsubstituted or substituted aryl and unsubstituted or substituted heteroaryl; y is selected from unsubstituted or substituted alkyl, or a 4-, 5-, 6-membered cycloalkyl ring; and R is₁Selected from H or lower alkyl; or, Y and R₁May be joined together to form a 4-, 5-or 6-membered heterocyclic ring; g is selected from the group consisting of H,

wherein R is₂、R₃And R₄Each independently selected from H, halogen, -COOH, unsubstituted or substituted lower alkyl, unsubstituted or substituted lower heteroalkyl; z is selected from halogen or hydroxyl; preferably, L₁Selected from unsubstituted or substituted cis or trans C2 alkenyl and unsubstituted or substituted C2 alkynyl; l is₂Is selected from- (CH)₂)_n-O-, wherein n is 0, 1 or 2; ar is selected from unsubstituted or substituted aryl;

is selected from

More preferably still, the first and second liquid crystal compositions are,

is selected from

G is selected from

Z is selected from chlorine.

The process for preparing compound I may be a conventional process in the art for such amide-forming reactions, and the following reaction processes and conditions are particularly preferred in the present invention: mixing the compound I-f with an organic solvent and a base and/or a condensing agent, and further reacting with a compound G-Z (unsubstituted or substituted lower alkane, carboxylic acid or acyl halide) to obtain a compound I;

in the method for preparing the compound I, the solvent is preferably one or more of an ether solvent, an alcohol solvent, a nitrile solvent, an alkane solvent and an aprotic polar solvent; further preferred is an alkane solvent. The ether solvent is preferably tetrahydrofuran. The alcohol solvent is preferably methanol and/or ethanol. The nitrile solvent is preferably acetonitrile. The alkane solvent is preferably dichloromethane. The aprotic polar solvent is preferably N, N-dimethylformamide.

In the process for producing the compound I, the volume-to-mass ratio of the solvent to the compound I-f is preferably 5mL/g to 50mL/g, and more preferably 10mL/g to 30 mL/g.

In the process for preparing compound I, the base is preferably an organic base, such as triethylamine, diisopropylethylamine, pyridine, lutidine, collidine, 4-dimethylaminopyridine, potassium tert-butyrate, sodium methoxide, sodium ethoxide, lithium hexamethyldisilazide, sodium hexamethyldisilazide, potassium hexamethyldisilazide and butyl lithium; and inorganic bases such as sodium hydrogen carbonate, sodium carbonate, potassium carbonate, cesium carbonate, sodium hydroxide and sodium hydride. Further preferred are triethylamine and diisopropylethylamine.

In the process for preparing compound I, the molar ratio of the base to the compound I-f is preferably 1:1 to 5:1, more preferably 2:1 to 3: 1.

In the process for producing the compound I, the condensing agent is preferably CDI, DCC, EDCI, DIC, HATU, HBTU, PyBOP or the like, and if necessary, an acylation catalyst such as DMAP, HOBt or the like may be added. EDCI and HATU are further preferred.

In the process for producing compound I, the molar ratio of the condensing agent to the compound G-Z is preferably 1:1 to 5:1, more preferably 1:1 to 3: 1.

In the process for producing compound I, the molar ratio of the acylation catalyst to the compound G-Z is preferably 0.001:1 to 5:1, and more preferably 0.1:1 to 2: 1.

In the process for preparing compound I, the molar ratio of said compound G-Z to said compound I-f is preferably 1:1 to 5:1, more preferably 1:1 to 3: 1.

In the process for preparing compound I, the temperature of the amide-forming reaction is preferably-30 ℃ to 150 ℃, and more preferably-10 ℃ to 30 ℃.

In the process for preparing the compound I, the progress of the amide-forming reaction can be monitored by a conventional test method in the art (e.g., TLC or HPLC), and generally, the time when the compound I-f disappears is used as the end point of the reaction, and the reaction time is preferably 0.1h to 30h, more preferably 0.1h to 16 h.

Preferably, the process for the preparation of compound I comprises any of the following work-up steps: when Z is selected from chlorine, the method (1) is preferably employed. When Z is selected from hydroxyl groups, the method (2) is preferably employed. The method (1) comprises the following steps: after the reaction is finished, adding water, washing an organic phase by using water, a citric acid aqueous solution, saturated sodium bicarbonate and saturated sodium chloride, drying, concentrating and carrying out column chromatography. The method (2) comprises the following steps: after the reaction is finished, adding an organic solvent and water, washing an organic phase by using water and saturated sodium chloride, drying, concentrating and carrying out column chromatography. Among them, the organic solvent is more preferably ethyl acetate or dichloromethane. The method and conditions of column chromatography are selected according to the methods and conditions of column chromatography which are conventional in the art.

Step E: synthesis of Compound 2: carrying out substitution reaction on the compound 1 and NIS in a solvent to obtain a compound 2;

the method for preparing compound 2 may be a conventional method for such substitution reaction in the art, and the following reaction method and conditions are particularly preferred in the present invention: mixing the compound 1 with an organic solvent, and performing substitution reaction with NIS to obtain a compound 2;

in the method for preparing the compound 2, the solvent is preferably one or more of an ether solvent, an alcohol solvent, a nitrile solvent, acetone, DMF and water; DMF is more preferable. The ether solvent is preferably tetrahydrofuran. The alcohol solvent is preferably methanol and/or ethanol. The nitrile solvent is preferably acetonitrile.

In the method for preparing the compound 2, the volume-to-mass ratio of the solvent to the compound 1 is preferably 5mL/g to 50mL/g, and more preferably 10mL/g to 30 mL/g.

In the process for preparing compound 2, the molar ratio of said compound 1 to said NIS is preferably 1:1 to 1:5, more preferably 1:1 to 1: 2.

In the process for producing compound 2, the temperature of the substitution reaction is preferably 0 ℃ to 150 ℃, and more preferably 50 ℃ to 100 ℃.

In the process for preparing compound 2, the progress of the substitution reaction can be monitored by a conventional test method in the art (e.g., TLC or HPLC), and generally the end point of the reaction is determined as the disappearance of compound 1, and the reaction time is preferably 0.5h to 16h, more preferably 5h to 12 h.

The following work-up step is preferred when the solvent is selected from DMF in the process for the preparation of compound 2: cooling to room temperature, adding water, filtering, sequentially washing with water and organic solvent, and drying at 50-90 deg.C. Among them, ethanol is more preferable as the organic solvent.

Step F: synthesis of I-d: in a solvent, compound 2 and compound 3 are subjected to a Mitsunobu reaction in the presence of a tri-substituted phosphine and a di-substituted azodicarboxylate.

Wherein Y in compound 3 and compounds I-d is selected from unsubstituted or substituted alkyl, or is a 4-, 5-, 6-membered cycloalkyl ring; and R is₁Selected from H or lower alkyl; or, Y and R₁Can be combined togetherTo form a 4-, 5-or 6-membered heterocyclic ring; preferably, the first and second electrodes are formed of a metal,

is selected from

More preferably still, the first and second liquid crystal compositions are,

is selected from

The method for preparing compounds I-d may be a conventional method in the art for such Mitsunobu reactions, and the following reaction methods and conditions are particularly preferred in the present invention: in a solvent, compound 2 and compound 3 are subjected to a Mitsunobu reaction in the presence of triphenylphosphine and diisopropyl azodicarboxylate.

The process for preparing compounds I-d preferably comprises the following steps: at 0-10 ℃, the diisopropyl azodicarboxylate is dropped into a solution formed by the compound 2, the triphenylphosphine and the compound 3 and a solvent to carry out the Mitsunobu reaction.

In the method for preparing the compounds I-d, the solvent is preferably one or more of nitrile solvents, ether solvents and halogenated hydrocarbon solvents; further preferred is an ether solvent. The nitrile solvent is preferably acetonitrile; the ether solvent is preferably tetrahydrofuran; the halogenated hydrocarbon solvent is preferably a chlorinated hydrocarbon solvent, and more preferably dichloromethane.

In the method for preparing the compounds I-d, the volume-to-mass ratio of the solvent to the compound 2 is preferably 1mL/g to 40mL/g, and more preferably 5mL/g to 20 mL/g.

In the process for producing the compounds I to d, the molar ratio of the diisopropyl azodicarboxylate to the compound 2 is preferably 1:1 to 3:1, more preferably 1:1 to 2: 1.

In the process for preparing the compounds I-d, the molar ratio of the triphenylphosphine to the compound 2 is preferably from 1:1 to 3:1, more preferably from 1:1 to 2: 1.

In the third process for preparing the compounds I-d, the molar ratio of the compound 3 to the compound 2 is preferably 1:1 to 3:1, and more preferably 1:1 to 2: 1.

In the process for producing the compounds I to d, the temperature of the Mitsunobu reaction is preferably from-10 ℃ to 35 ℃, and more preferably from 0 ℃ to 20 ℃.

In the process for preparing the compounds I-d, the progress of the Mitsunobu reaction can be monitored by a conventional test method in the art (e.g., TLC or HPLC), and generally, the reaction time is preferably 0.5h to 24h, more preferably 3h to 12h, with the disappearance of the compounds I-d as the end point of the reaction.

The process for preparing compounds I-d preferably comprises the following work-up steps: and after the reaction is finished, evaporating the solvent under reduced pressure, and purifying by column chromatography. The method and conditions of column chromatography are selected according to the methods and conditions of column chromatography which are conventional in the art.

The synthesis of the compounds of formula (I) is outlined in the examples.

Further forms of the compounds

In a certain embodiment, the compounds of formula (I) are prepared according to a pharmaceutically acceptable acid addition salt (a pharmaceutically acceptable salt) by reacting the free base form of the compound with a pharmaceutically acceptable inorganic or organic acid, including but not limited to inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, hydrogen phosphate, sulfuric acid, hydrogen sulfate and the like; organic acids formic acid, acetic acid, trifluoroacetic acid, malic acid, maleic acid, fumaric acid, succinic acid, benzoic acid, methanesulfonic acid, benzenesulfonic acid, acetic acid, propionic acid, butyric acid, octanoic acid, adipic acid, oxalic acid, malonic acid, succinic acid, tartaric acid, phenylacetic acid, phthalic acid, terephthalic acid, valeric acid, hexanoic acid, decanoic acid, stearic acid, palmitic acid, acrylic acid, and the like.

The inorganic acid is preferably selected from hydrochloric acid, hydrobromic acid or phosphoric acid. The organic acid is preferably selected from formic acid, acetic acid, trifluoroacetic acid, malic acid, maleic acid, fumaric acid, succinic acid, benzoic acid, methanesulfonic acid or benzenesulfonic acid; more preferably from acetic acid or trifluoroacetic acid.

"pharmaceutically acceptable" as used herein refers to a substance, such as a carrier or diluent, which does not diminish the biological activity or properties of the compound and which is relatively non-toxic, e.g., by being administered to an individual without causing unwanted biological effects or interacting in a deleterious manner with any of the components it contains.

The term "pharmaceutically acceptable salt" refers to a form of a compound that does not cause significant irritation to the organism to which it is administered and does not abrogate the biological activity and properties of the compound. In certain particular aspects, pharmaceutically acceptable salts are obtained by reacting a compound of formula (I) with an acid, such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid and the like. Pharmaceutically acceptable salts may also be formed by reacting a compound of formula (I) with a base to form a salt, such as an ammonium salt; alkali metal salts, such as sodium or potassium salts; alkaline earth metal salts, such as calcium or magnesium salts; organic base salts such as dicyclohexylamine, N-methyl-D-glucamine, tris (hydroxymethyl) methylamine; amino acid salts such as arginine, lysine and the like.

References to pharmaceutically acceptable salts are understood to include solvent addition forms or crystalline forms, especially solvates or polymorphs. Solvates contain either stoichiometric or non-stoichiometric amounts of solvent and are selectively formed during crystallization with pharmaceutically acceptable solvents such as water, ethanol, and the like. Hydrates are formed when the solvent is water, or alcoholates are formed when the solvent is ethanol. Solvates of the compounds of formula (I) are conveniently prepared or formed as described herein. Illustratively, hydrates of the compounds of formula (I) are conveniently prepared by recrystallization from a mixed solvent of water/organic solvent, using organic solvents including, but not limited to, dioxane, tetrahydrofuran, ethanol or methanol. In addition, the compounds mentioned herein can exist in unsolvated as well as solvated forms. In general, the solvated forms are considered equivalent to unsolvated forms for purposes of the compounds and methods provided herein.

In other embodiments, the compounds of formula (I) are prepared in different forms, including, but not limited to, amorphous, pulverized, and nano-sized forms. In addition, the compounds of formula (I) include crystalline forms, as well as polymorphic forms. Polymorphs include different lattice arrangements of the same elemental composition of a compound. Polymorphs typically have different X-ray diffraction patterns, infrared spectra, melting points, densities, hardness, crystal forms, optical and electrical properties, stability and solubility. Different factors such as recrystallization solvent, crystallization rate and storage temperature may cause a single crystal form to dominate.

In certain embodiments, the compounds of formula (I) are prepared as prodrugs. By "prodrug" is meant an agent that is converted in vivo to the proto-drug. Prodrugs are often useful because, in some cases, they may be easier to administer than the proto-drug. They can, for example, be bioavailable by oral administration, but prototype drugs are not. Prodrugs can also improve the solubility of the proto-drug in the pharmaceutical composition. For example, without limitation, prodrugs are compounds of formula (I) and, where water solubility is not conducive to passage through a cell membrane, prodrugs are administered as esters to facilitate passage through a cell membrane, and then hydrolyzed metabolically to carboxylic acids, which are advantageous once the active entity has entered the cell. As a further example, a prodrug may be a short peptide (polyamino acid) linked to an acid group, the peptide being metabolized to reveal the active fragment.

Prodrugs are generally precursors to drugs which, following administration and absorption, are converted to the active species or, by some process, are converted to more active species, such as by metabolic pathways. Some prodrugs have chemical groups that make them less active and/or less soluble than the proto-drug or some other property. Once the chemical groups of the prodrug are removed and/or modified, the active drug is obtained. Prodrugs are often useful, and in some cases they are easier to administer than the proto-drug. In certain embodiments, the prodrug compounds described herein are bioavailable by oral administration, but the proto-drugs are not. Moreover, in certain embodiments, the prodrugs described herein may also improve the solubility of the proto-drug in pharmaceutical compositions.

In other embodiments, the prodrug is designed as a reversible drug derivative, used as a modifier to enhance drug transport to tissues at a specific location. In a particular aspect, the prodrug design is aimed at increasing the effective water solubility of the therapeutic compound for which the targeted region is water as the predominant solvent. Fedorak et al, am.J.Physiol., 269: G210-218 (1995); McLoed et al, Gastroenterol, 106: 405-; hochhaus et al, biomed.Chrom, 6: 283-; larsen and h.bundgaard, int.j.pharmaceuticals, 37, 87 (1987); j.larsen et al, int.j.pharmaceuticals, 47, 103 (1988); sinkula et al, J.Pharm.Sci, 64:181-210 (1975); t.higuchi and v.stella, produgs as Novel Delivery Systems, vol.14 of the a.c.s.symposium Series; and Edward B.Roche, Bioreversible Carriers in Drug Design, American Pharmaceutical Association and Pergamon Press, 1987.

In another embodiment, the compounds described herein are isotopically labeled (e.g., radioisotopes) or labeled by other means, including, but not limited to, the use of chromophores or fluorescent moieties, luminescent labels, or chemiluminescent labels.

In another aspect, the compounds of formula (I) have one or more stereogenic centers, each center independently present in the R or S configuration. The compounds mentioned herein include all diastereomers, enantiomers, epimers and suitable mixtures thereof. In a particular aspect, the compounds of formula (I) are prepared as their single stereoisomers by reacting a racemic mixture with an optically active resolving agent to form a pair of diastereomers, and separating the diastereomers to give the optically pure enantiomers. In certain particular aspects, separation of the enantiomers is performed using covalent diastereomeric derivatives of the compounds described herein. In other particular aspects, isolatable complexes (e.g., crystalline diastereomeric salts) are used. Diastereomers have different physical properties (e.g., melting points, boiling points, solubilities, reactivities, etc.), and in particular aspects, diastereomers can be separated by taking advantage of these different properties. In these particular aspects, the diastereomers are separated by chiral chromatography or separation techniques based on differences in solubility. Optically pure enantiomers are obtained by any feasible method that does not result in racemization, along with resolving agents. Jean Jacques, Andre Collet, Samuel H.Wilen, "Enantiomers, Racemates And solutions," John Wiley And Sons, Inc., 1981.

In addition, in certain embodiments, the compounds provided herein exist as geometric isomers. The compounds and methods provided herein include all cis, trans, E, Z isomers and suitable mixtures thereof. In certain particular aspects, the compounds described herein exist as tautomers. All tautomers are within the molecular structural formulas described herein. In further aspects of the compounds and methods provided herein, mixtures of enantiomers and/or diastereomers resulting from a single preparative step (combination or interconversion) are contemplated.

Therapeutic uses

In one aspect, the compounds of formula I inhibit Btk. In another aspect, the compounds of formula I exhibit antiproliferative activity and are effective in the treatment of proliferative diseases. In one aspect, the compounds of formula I show utility in the prevention or treatment of inflammation, autoimmune diseases associated with abnormal B cell proliferation and neoplastic diseases.

In a further aspect, provided herein is a method of inhibiting bruton's tyrosine kinase activity in a subject by administering to a subject in need thereof a composition comprising a therapeutically effective amount of at least one compound or composition of formula I. In some embodiments, a subject in need thereof is suffering from an autoimmune disease, such as inflammatory bowel disease, arthritis, lupus, rheumatoid arthritis, psoriatic arthritis, osteoarthritis, still's disease, juvenile arthritis, rheumatoid arthritis syndrome, autoimmune hepatitis, and the like.

In further embodiments, the subject in need thereof is suffering from cancer. In one embodiment, the cancer is a B cell proliferative disease, such as diffuse large B cell lymphoma, follicular lymphoma, chronic lymphocytic leukemia, B cell prolymphocytic leukemia, lymphoplasmacytic lymphoma/waldenstrom macroglobulinemia, splenic marginal zone lymphoma, plasmacytic myeloma, plasmacytoma, extranodal marginal zone B cell lymphoma, lymph node marginal zone B cell lymphoma, mantle cell lymphoma, mediastinal (thymic) large B cell lymphoma, intravascular large B cell lymphoma, primary effusion lymphoma, burkitt lymphoma/leukemia, or lymphomatoid granulomatosis, and the like.

The invention also provides application of the compound or the medicinal salt thereof in preparing Btk inhibitors, in particular application in preparing drugs for treating cell proliferative diseases. The cell proliferative disease includes cancer. In other words, the invention also provides the application of the compound shown in the general formula I or the pharmaceutically acceptable salt or solvate thereof in treating proliferative diseases (such as cancer) alone or in combination with other medicines. Antineoplastic agents that can be used in combination with the compounds provided herein or pharmaceutically acceptable salts thereof include, but are not limited to, at least one of the following: mitotic inhibitors (e.g., vinblastine, vindesine, and vinorelbine); tubulysin decomposition inhibitors (e.g., taxol); alkylating agents (such as cisplatin, carboplatin, and cyclophosphamide); antimetabolites (e.g., 5-fluorouracil, tegafur, methotrexate, cytarabine, and hydroxyurea); antibiotics (such as adriamycin, mitomycin and bleomycin) can be inserted; enzymes (e.g., asparaginase); topoisomerase inhibitors (e.g., etoposide and camptothecin) and biological response modifiers (e.g., interferons), among others.

The compound of the invention is used for preparing a preparation, which comprises the following steps: the compound is used directly or as any one of the components obtained during the preparation process. The invention also provides a pharmaceutical composition comprising a certain amount of a compound of formula I capable of inhibiting Btk activity and a pharmaceutically acceptable excipient. The pharmaceutical composition is in the form of aqueous dispersion, liquid, gel, syrup, elixir, syrup, suspension, inhalant, controlled-release agent, quick-dissolving agent, effervescent agent, lyophilized agent, tablet, powder, pill, dragee, capsule, delayed-release agent, extended-release agent, pulsatile release agent, multiparticulate, or immediate-release agent.

The pharmaceutical compositions described herein consist of a pharmaceutically acceptable diluent, excipient, or binder and a compound of formula I, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.

In one aspect, the pharmaceutical compositions described herein comprise an effective amount of a compound as described above and a pharmaceutically acceptable excipient. In another aspect, the pharmaceutical composition comprises, in addition to the compound of formula I, a second pharmaceutically active ingredient.

In a particular embodiment, there is provided a pharmaceutical composition comprising: i) a physiologically acceptable carrier, diluent and/or excipient; and ii) a compound as set forth herein.

Any of the foregoing aspects contains further embodiments that include the mere administration of an effective amount of a compound of formula I. These preferred examples specifically include: i) administering a compound of formula I in one dose; ii) administering the compound of formula I to the mammal multiple times during the day; iii) continuous administration of a compound of formula I; or iv) continuous administration of the compound of formula I.

Any of the above aspects contains further embodiments comprising multiple administrations of an effective amount of a compound of formula I. These preferred examples specifically include: i) administering a compound of formula I in a single dose; ii) multiple administrations at six hour intervals; iii) administering the compound of formula I to the mammal every eight hours. In a further or alternative preferred embodiment, the method has a drug holiday, in particular a temporary delay in the administration of the compound of formula I or a temporary reduction in the dose of the compound of formula I; at the end of the drug holiday, the dosage of the compound of formula I is resumed. The time of drug holidays varies from two days to one year.

The compound of the invention can be prepared into a pharmaceutical composition with various common additives (such as diluent, excipient and the like) in pharmacy. The pharmaceutical composition may be formulated into various types of administration unit dosage forms such as tablets, pills, powders, liquids, suspensions, emulsions, granules, capsules, suppositories, injections (solutions and suspensions) and the like, depending on the purpose of treatment.

For shaping the pharmaceutical composition in tablet form, any excipient known and widely used in the art may be used. For example, carriers such as lactose, white sugar, sodium chloride, glucose, urea, starch, calcium carbonate, kaolin, crystalline cellulose, silicic acid, and the like; binders such as water, ethanol, propanol, common syrup, glucose solution, starch solution, gelatin solution, carboxymethyl cellulose, shellac, methyl cellulose and potassium phosphate, polyvinylpyrrolidone, etc.; disintegrators such as dry starch, sodium alginate, agar powder and kelp powder, sodium bicarbonate, calcium carbonate, fatty acid esters of polyethylene sorbitan, sodium lauryl sulfate, monoglyceride stearate, starch, lactose and the like; disintegration inhibitors such as white sugar, glycerol tristearate, coconut oil and hydrogenated oil; adsorption promoters such as quaternary ammonium bases and sodium lauryl sulfate, etc.; humectants such as glycerin, starch, and the like; adsorbents such as starch, lactose, kaolin, bentonite, colloidal silicic acid, and the like; and lubricants such as pure talc, stearates, boric acid powder, polyethylene glycol, and the like. If desired, the tablets can also be made as sugar-coated tablets, gelatin-coated tablets, enteric-coated tablets, film-coated tablets, double-layer tablets and multilayer tablets with the usual coating materials.

For shaping the pharmaceutical composition in the form of a pill, any of the excipients known and widely used in the art may be used, for example, carriers such as lactose, starch, coconut oil, hardened vegetable oil, kaolin, talc and the like; adhesives such as gum arabic powder, xanthan gum powder, gelatin, ethanol, and the like; disintegrating agents, such as agar and kelp powder.

For shaping the pharmaceutical composition in the form of suppositories, any excipient known and widely used in the art may be used, for example, polyethylene glycol, coconut oil, higher alcohols, esters of higher alcohols, gelatin, semisynthetic glycerides and the like.

For the preparation of pharmaceutical compositions in the form of injection solutions, the solutions and suspensions may be sterilized and, preferably, suitable amounts of sodium chloride, glucose or glycerol, etc., may be added to prepare an injection solution which is isotonic with blood. In the preparation of injection, any carrier commonly used in the art may also be used. For example, water, ethanol, propylene glycol, ethoxylated isostearyl alcohol, polyoxylated isostearyl alcohol, and fatty acid esters of polyethylene sorbitan, and the like. In addition, conventional lytic agents, buffers, analgesics, and the like may be added. Coloring agents, preservatives, perfumes, flavoring agents, perfuming agents and other medicines may also be added as required during the treatment of schizophrenia.

The content of the compound shown in the formula I and the pharmaceutically acceptable salt thereof in the pharmaceutical composition is not particularly limited, and can be selected within a wide range, and generally can be 1-90% by mass, and preferably 1-30% by mass.

In the present invention, the method of administration of the pharmaceutical composition is not particularly limited. The formulation of various dosage forms can be selected for administration according to the age, sex and other conditions and symptoms of the patient. For example, tablets, pills, solutions, suspensions, emulsions, granules and capsules are administered orally; the injection can be administered alone, or mixed with injectable delivery solution (such as glucose solution and amino acid solution) for intravenous injection, or simply injected into muscle, skin or abdomen if necessary; the suppository is administered to the rectum.

In the present invention, the administration dose can be appropriately selected depending on the administration method, the age, sex and other conditions of the patient and the symptoms.

The positive progress effects of the invention are as follows:

the pyrazolo [3,4-d ] pyrimidine compound and the salt thereof have good inhibition effect on Btk, partial compound activity/water solubility is superior to that of a marketed drug ibrutinib, diversified structures are provided for research and development of Btk small molecule inhibitors, and the compound has good market development prospect.

The above preferred conditions can be arbitrarily combined to obtain preferred embodiments of the present invention without departing from the common general knowledge in the art.

The invention is further illustrated by the following examples, which are not intended to limit the scope of the invention. Experimental procedures without specific conditions noted in the following examples were selected according to conventional procedures and conditions, or according to commercial instructions. Without further elaboration, it is believed that one skilled in the art can, based on the description herein, utilize the present disclosure to its fullest extent.

In the following examples, undefined abbreviations have their commonly accepted meaning unless otherwise stated. Table 1, Table 2 and Table 3 show the physicochemical data of Compound I-1 to I-24 and the physicochemical data of Compound I-1 to I-24¹H-NMR data and I-1-I-24 in vitro enzyme level inhibitory activity data.

Example 1

Preparation of Key intermediate 4

Step A: synthesis of Compound 2

Compound 1(15g, 111mmol), NIS (30g, 133.2mmol) and DMF (150ml) were stirred at 90 ℃ for 4h, then cooled to room temperature, added with water 150ml, the solid was filtered, washed with water 100ml, ethanol 100ml in sequence, dried at 70 ℃ to obtain a solid 23g, yield: 79.4 percent.

And B: synthesis of Compound 4

Compound 2(15g, 57.47mmol), (S) -1-tert-butoxycarbonyl-3-hydroxypiperidine (23.2g, 114.9mmol), triphenylphosphine (30.1g, 114.9mmol) and 150ml THF were cooled to 0 ℃ in an ice salt bath, DIAD (23.2g, 114.9mmol) was added dropwise, warmed to room temperature and stirred overnight, and column chromatography gave 13.1g of a solid, yield: 51.3 percent.

Example 2

Preparation of Compound I-1

Step A: synthesis of Compound 5

Bromopropyne (18.9g, 159mmol), phenol (10g, 106mmol), potassium carbonate (44g, 318mmol) were stirred at room temperature overnight in 100ml of DMF, ethyl acetate was added, and the mixture was washed with saturated sodium bicarbonate, saturated sodium chloride, and water in this order, and concentrated to give 11.9g of a transparent oily liquid, with a yield of 84.3%.

And B: synthesis of Compound 6

Compound 4(4.45g, 10mmol), CuI (0.38g, 2mmol), Pd (PPh)₃)₄(1.39g，1.2mmol)，Et₃N (2.03g, 20mmol), compound 5(6.62g, 50mmol) and 50ml DMF were warmed to 80 ℃ under nitrogen, stirred overnight, cooled to room temperature, and concentrated by column chromatography to give a solid 3.31g, yield: 73.6 percent.

And C: synthesis of Compound 7

Compound 6(3.3g, 10.5mmol), 4M HCl-1,4-dioxane (20ml, 72mmol) and 20ml ethyl acetate were stirred overnight at room temperature, filtered and dried at 75 ℃ to give a solid 1.9g, yield: 67.1 percent.

Step D: synthesis of Compound I-1

Compound 7(600mg, 1.56mmol), DIPEA (403mg, 3.12mmol) and 30ml DCM were cooled in an ice salt bath, 10ml of a DCM solution of acryloyl chloride (183mg, 2.03mmol) was added dropwise, warmed to room temperature, concentrated and subjected to column chromatography to give a solid 420mg, yield: 67%.

Example 3

Preparation of Compound I-2

The synthesis of compound I-2 was accomplished by using procedures analogous to those described in example 2.

Example 4

Preparation of Compound I-3

The synthesis of compound I-3 was accomplished by using procedures analogous to those described in example 2.

Example 5

Preparation of Compound I-4

The synthesis of compound I-4 was accomplished by using procedures analogous to those described in example 2.

Example 6

Preparation of Compound I-5

The synthesis of compound I-5 was accomplished by using procedures analogous to those described in example 2.

Example 7

Preparation of Compound I-6

The synthesis of compound I-6 was accomplished using procedures analogous to those described in example 2.

Example 8

Preparation of Compound I-7

The synthesis of compound I-7 was accomplished by using procedures analogous to those described in example 2.

Example 9

Preparation of Compound I-8

The synthesis of compound I-8 was accomplished using procedures analogous to those described in example 2.

Example 10

Preparation of Compound I-9

The synthesis of compound I-9 was accomplished by using procedures analogous to those described in example 2.

Example 11

Preparation of Compound I-10

The synthesis of compound I-10 was accomplished using procedures analogous to those described in example 2.

Example 12

Preparation of Compound I-11

The synthesis of compound I-11 was accomplished using procedures analogous to those described in example 2.

Example 13

Preparation of Compound I-12

The synthesis of compound I-12 was accomplished using procedures analogous to those described in example 2.

Example 14

Preparation of Compound I-13

The synthesis of compound I-13 was accomplished using procedures analogous to those described in example 2.

Example 15

Preparation of Compound I-14

The synthesis of compound I-14 was accomplished using procedures analogous to those described in example 2.

Example 16

Preparation of Compound I-15

The synthesis of compound I-15 was accomplished using procedures analogous to those described in example 2.

Example 17

Preparation of Compound I-16

Step A: synthesis of Compound 8

3-butyn-1-ol (10g, 142.67mmol), phenol (16.11g, 171.2mmol), triphenylphosphine (44.9g, 171.2mmol) and 100ml THF were cooled in an ice salt bath, DIAD (34.62g, 171.2mmol) was added dropwise at 0 deg.C, then warmed to room temperature and stirred overnight, and column chromatography gave 5.45g of liquid, yield: 26.1 percent.

And B: the synthesis of compounds 9, 10 and I-16 was accomplished using procedures analogous to those described in example 2.

Example 18

Preparation of Compound I-17

Step A: synthesis of Compound 11

Compound 7(880mg, 2.3mmol), 10% Pd-BaSO4(260mg, 0.23mmol) and 20ml methanol were stirred under normal pressure hydrogen at 45 ℃ overnight, after which column chromatography gave 470mg of a solid, yield: 60 percent.

And B: the synthesis of compound I-17 was accomplished using procedures analogous to those described in example 2.

Example 19

Preparation of Compound I-18

Step A: synthesis of Compound 12

Compound 7(1.57g, 4.08mmol) and 70ml THF were added to LiAlH at 0 deg.C₄(929mg, 24.47mmol), warmed to room temperature and stirred overnight, ethyl acetate and aqueous sodium hydroxide solution were added, and column chromatography was performed to obtain a solid 330mg, yield: 23.2 percent.

And B: the synthesis of compound I-18 was accomplished using procedures analogous to those described in example 2.

Example 20

Preparation of Compound I-19

Step A: synthesis of Compound 13

Compound 7(1g, 2.6mmol), 10% Pd/C (400mg) and 30ml methanol were stirred under normal pressure hydrogen at 55 ℃ overnight, filtered and column chromatographed to give 890mg of oil, yield: 88.08 percent.

And B: the synthesis of compound I-19 was accomplished using procedures analogous to those described in example 2.

Example 21

Preparation of Compound I-20

Step A: synthesis of Compound 14

Adding i-PrMgCl (2M in THF 117ml, 233.88mmol) into trimethylacetylene silicon (23g, 233.88mmol) and 100ml THF at 0 ℃, stirring for 30min, raising the temperature to room temperature, stirring for 30min, adding CuBr (5.03g, 35.08mmol) and benzyl bromide (10g, 58.47mmol), refluxing for 5h, cooling to room temperature, adding ethyl acetate, washing with water, and performing column chromatography to obtain a liquid 9.2g, wherein the yield is as follows: 83.5 percent.

And B: synthesis of Compound 15

Compound 14(9.2g, 48.85mmol) and 100ml methanol were cooled in an ice bath, potassium carbonate (33.76g, 244.2mmol) was added and stirred for 2h, stirred overnight at room temperature, ethyl acetate was added, washed with water and concentrated to give 5g of oil which was directly charged to the next step.

And C: the synthesis of compounds 16, 17 and I-20 was accomplished using procedures analogous to those described in example 2.

Example 22

Preparation of Compound I-21

2-Fluoroacrylic acid (175mg, 1.95mmol) and 10ml DMF were added DIPEA (504mg, 3.9mmol), HATU (741mg, 1.95mmol) and Compound 7(500mg, 1.3mmol) at 0 deg.C, stirred overnight at room temperature, added ethyl acetate, washed with water, and column chromatographed to give 330mg of a solid, yield: 60 percent.

Example 23

Preparation of Compound I-22

Step A: the synthesis of compound 18 was accomplished using step B analogous to that described in example 1.

And B: the synthesis of compounds 19, 20 and I-22 was accomplished using procedures analogous to those described in example 2.

Example 24

Preparation of Compound I-23

Step A: the synthesis of compound 21 was accomplished using step B analogous to that described in example 1.

And B: the synthesis of compounds 22, 23 and I-23 was accomplished using procedures analogous to those described in example 2.

Example 25

Preparation of Compound I-24

Step A: the synthesis of compound 24 was accomplished using a procedure similar to step B described in example 1.

And B: the synthesis of compounds 25, 26 and I-24 was accomplished using procedures analogous to those described in example 2.

The room temperature in the invention refers to the ambient temperature and is 15-30 ℃.

The reagents and starting materials used in the present invention are commercially available.

TABLE 2 physicochemical data for Compounds I-1-I-24

TABLE 3 preparation of compounds I-1-I-24¹H-NMR data

Example 26

Btk inhibitory Activity assay of Compounds at Biochemical enzyme level in vitro

The principle and the method are as follows:

a kinase activity detection platform of the Btk small molecule inhibitor is established by adopting a homogeneous phase time-resolved fluorescence (HTRF) method, and the enzyme level activity of the compound is measured. The method for measuring the activity of the Btk inhibitor enzyme is characterized in that biotin is connected to the other end of a monophosphonization site substrate which can react with TK kinase, and the biotin can be connected with streptavidin-XL665 (receptor) at the biotin end to finish marking. After phosphorylation, the substrate binds to an antibody (labeled Doner) against the phosphorylation site, resulting in a FRET signal, as shown in fig. 1.

The method comprises the following operation steps:

1. test compounds were dissolved in 100% DMSO at 100 μ M initial dilution and then diluted in 96-well plates with a configured buffer gradient.

2. Transfer 4 μ l of diluted compound to 384 well plates.

3. A buffer of substrate and ATP was prepared.

4. Mu.l of substrate and ATP buffer were added to 384-well plates using a microplate applicator.

5. Mu.l of enzyme buffer was added to the 384-well plate using a microplate applicator.

6. The 384 well plates were gently shaken, centrifuged at 1000g for a short time at room temperature, and incubated for 60 minutes at room temperature.

7. The stop solutions were prepared, and 5. mu.l each of SA-XL665 and TK antibody was added to the solutions using a microplate applicator, shaken for 1 minute, centrifuged, and incubated at room temperature for 60 minutes. See figure 2.

Reading by a PHERAStar multifunctional plate reader.

GraphPad Prism 5.0 analytical software analysis data.

Solving IC by the column equation₅₀：

Computing IC₅₀The value:

in log [ administration concentration ]]The abscissa and the ordinate represent the inhibition rate, and a dose-response curve is fitted to GraphPad Prism 5.0 to obtain the drug concentration at 50% inhibition rate, i.e., the IC of the compound at the enzyme level₅₀The value is obtained.

The compound Btk inhibitory activity data are as follows:

compound number	Btk IC50(nM)	Numbering	Btk IC50(nM)
				I-1	2.95	I-14	209.2
I-2	18.1	I-15	45.35
				I-3	3.95	I-16	39.8
I-4	15.9	I-17	23.2
				I-5	>500	I-18	11.3
I-6	>500	I-19	22.18
				I-7	49.23	I-20	102.4
I-8	20.85	I-21	>500
				I-9	26.89	I-22	>1000
I-10	38.17	I-23	>1000
				I-11	3.84	I-24	>1000
I-12	89.62	Ibrutinib	5.49
				I-13	18.11

The results show that the synthesized compound has excellent Btk inhibitory activity. The activity of part of compounds disclosed in the formula I-1, I-3 and I-11 is superior to that of Ibrutinib, and the formula plays a guiding role in developing Btk small molecule inhibitors with diversified structures and modifying the structures in the future.

Example 27

Comparison of Btk inhibitory Activity at Biochemical enzyme level in vitro

The structural formula of the compound A described in Chinese patent application CN103958512A is as follows:

the compound is an FGFR inhibitor, compound A is synthesized by a method reported in reference CN 103958512A. the in vitro BTK enzyme level inhibition activity of the compound A is tested by the method of example 26, and the results are as follows:

compound number	Btk IC50(nM)
		A	>1000

The results show that the activity of the compound provided by the invention for inhibiting Btk is far higher than that of the compound A.

Example 28

Test of BTK inhibitory Activity at the Compound in vitro cellular level

The method comprises the following steps: CCK-8 detection method

The method comprises the following operation steps:

1. 100ul of cell suspension (2000-₂) Preculture was carried out for 24 hours.

2. A gradient of test compound is added to the plate.

3. The plates were incubated in an incubator for 48 hours.

4. Add 10ul of CCK-8 solution to each well and shake the plate gently after addition.

5. The plates were incubated in an incubator for 1-4 hours.

6. Absorbance at 450nm was measured with a microplate reader.

The results are recorded:

cell growth inhibition ═ 100% control absorbance value-experimental absorbance value/control absorbance value;

computing IC₅₀The value:

in log [ administration concentration ]]On the abscissa and the ordinate, a dose-response curve was fitted to GraphPad Prism 5.0 to determine the drug concentration at 50% inhibition, i.e., the IC of the compound at the cellular level₅₀The value is obtained.

The activity results are shown below:

remarking: ramos and Raji are typical B-lymphocyte leukemia cells, highly expressed in BTK kinase.

The results show that most of the compounds provided by the invention have stronger inhibition effect on lymphocyte leukemia cells (Ramos and Raji), wherein the compound I-1 shows cell level inhibition activity equivalent to that of ibrutinib, so that the molecules are indicated to have the potential of being developed into novel high-efficiency BTK inhibitors, and the compounds have great application value in treating related tumor diseases, particularly diffuse large B cell lymphoma or chronic lymphocyte leukemia.

Example 29

cLog P values for some of the compounds

The cLog P value was calculated by ACD-Labs (V6.0).

Compound (I)	cLog P
		I-1	1.84±1.49
I-3	2.41±1.49
		I-4	2.42±1.49
I-11	1.64±1.49
		I-18	1.53±1.14
Ibrutinib (Ibrutinib)	2.92±1.18

The result shows that the solubility of the compound provided by the invention in water is better than that of ibrutinib, so that the dosage can be reduced, and the side effect can be reduced.

The embodiments described herein are for illustrative purposes only and various modifications or changes in light thereof will be suggested to persons skilled in the art and are to be included within the spirit and purview of this application and scope of the appended claims.

Claims (10)

1. A compound, or a pharmaceutically acceptable salt thereof, selected from the group consisting of:

2. a pharmaceutical composition comprising a compound of claim 1 and a pharmaceutically acceptable excipient.

3. The pharmaceutical composition as set forth in claim 2, wherein the pharmaceutical composition is in the form of an aqueous dispersion, liquid, gel, aerosol, controlled-release agent, fast-dissolving agent, effervescent agent, lyophilized agent, tablet, powder, pill, capsule, or multiparticulate.

4. The pharmaceutical composition of claim 3, wherein the controlled release agent is selected from a delayed release agent, an extended release agent, a pulsed controlled release agent, or an immediate release agent.

5. The pharmaceutical composition of claim 3, wherein the liquid is selected from a slurry, or a suspension.

6. The pharmaceutical composition of claim 5, wherein the slurry is a syrup.

7. The pharmaceutical composition of claim 3, wherein the pill is a sugar-coated pill.

8. A process for the preparation of a compound according to claim 1, comprising the steps of:

(1) mixing the compound 1, NIS and DMF for reaction to obtain a compound 2;

(2) mixing the compound 2, (S) -1-tert-butyloxycarbonyl-3-hydroxypiperidine, triphenylphosphine and DIAD for reaction to obtain a compound 4;

(3) mixing the compound 4 and the compound 5 for reaction to obtain a compound 6;

(4) mixing and reacting the compound 6, HCl and 1,4-dioxane to obtain a compound 7;

(5) mixing the compound 7, DIPEA and acryloyl chloride for reaction to obtain a solid compound I-1;

employing the above steps (1) to (5) in the case of substitution with the corresponding alkynyl compound to give compound I-3;

compound 7, THF and LiAlH₄Mixing and reacting to obtain a compound 12; mixing the compound 12, DIPEA and acryloyl chloride for reaction to obtain a solid compound I-18;

9. use of a compound according to claim 1 for the preparation of a medicament for the prevention or treatment of inflammation, autoimmune diseases associated with abnormal B-cell proliferation and/or neoplastic diseases.

10. The use of claim 9, wherein the autoimmune disease is rheumatoid arthritis.

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