WO2016180334A1 - Dual-site irreversible bruton's tyrosine kinase inhibitor, composition and application therefof - Google Patents

Dual-site irreversible bruton's tyrosine kinase inhibitor, composition and application therefof Download PDF

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Publication number
WO2016180334A1
WO2016180334A1 PCT/CN2016/081669 CN2016081669W WO2016180334A1 WO 2016180334 A1 WO2016180334 A1 WO 2016180334A1 CN 2016081669 W CN2016081669 W CN 2016081669W WO 2016180334 A1 WO2016180334 A1 WO 2016180334A1
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Prior art keywords
piperididine
compound
arh
pharmaceutically acceptable
tyrosine kinase
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PCT/CN2016/081669
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French (fr)
Chinese (zh)
Inventor
周星露
刘兴国
戈震
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杭州和正医药有限公司
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Priority claimed from CN201510242552.8A external-priority patent/CN104844609B/en
Priority claimed from CN201610286399.3A external-priority patent/CN105777759B/en
Application filed by 杭州和正医药有限公司 filed Critical 杭州和正医药有限公司
Publication of WO2016180334A1 publication Critical patent/WO2016180334A1/en

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/519—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim ortho- or peri-condensed with heterocyclic rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D487/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00
    • C07D487/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00 in which the condensed system contains two hetero rings
    • C07D487/04—Ortho-condensed systems

Definitions

  • the invention belongs to the field of medicine, in particular to a two-site irreversible Bruton tyrosine kinase inhibitor, a composition and application thereof.
  • Covalent inhibitors also known as irreversible inhibitors, are a class of inhibitors that exert their biological functions by irreversible binding of covalent bonds to target protein residues.
  • Covalent inhibitor drugs have made important contributions to human health over the past few decades. Covalent inhibitors enhance affinity to the target by covalent bonding to the target protein relative to the non-covalent inhibitor, which is the underlying cause of the high bioactivity of the covalent inhibitor.
  • due to the non-covalent targeting of anti-tumor drugs, especially the production of a large number of tini-resistant drugs against kinases people have paid more attention to covalent inhibitor drugs.
  • many large pharmaceutical companies have developed covalent inhibitors for specific enzyme targets.
  • afatinib was officially approved by the US FDA on July 12, 2013 for the treatment of metastatic non-small cell lung cancer with epidermal growth factor receptor (EGFR) gene mutation, becoming the first FDA-approved treatment of lung cancer.
  • EGFR epidermal growth factor receptor
  • New drug for irreversible inhibitors has also been a research hotspot in recent years, and great progress has been made.
  • the FDA approved two anti-hepatitis C virus covalent inhibitor drugs, namely, telaprevir. (Telaprevir) and Boceprevir (Boceprevir).
  • Btk Bruton's tyrosine kinase
  • BCR cell surface B-cell receptor
  • Btk is a key regulator of B cell development, activation, signaling, and survival.
  • Bkt plays a role in numerous other hematopoietic signaling pathways, such as Toll like receptor (TLR) and cytokine receptor-mediated TNF- ⁇ production in macrophages, in mast cells.
  • TLR Toll like receptor
  • TNF- ⁇ cytokine receptor-mediated TNF- ⁇ production in macrophages, in mast cells.
  • Immunoglobulin E receptor (Fc ⁇ R1) signaling signaling that inhibits apoptosis of Fas/APO-1 cells in B-lineage lymphoid cells, and collagen-stimulated platelet aggregation.
  • Fc ⁇ R1 Immunoglobulin E receptor
  • Btk inhibitors inhibit Btk autophosphorylation by binding to Btk by acting on the BCR signaling pathway, preventing Btk activation, thereby blocking cell conduction and inducing apoptosis.
  • Btk inhibitors have strong selectivity and low toxic and side effects, especially the listing of ibufenib, which has been designated as a “breakthrough” new drug by the FDA, and its research and development prospects are broad.
  • Ibbutinib reacts with the sulfhydryl group of the Btk enzyme cysteine (Cys481) residue and forms a covalent bond, which inactivates the Btk enzyme and exerts its therapeutic effect.
  • ibbutinibinib is easily metabolized during metabolism (digested by metabolic enzymes to be dihydroxylated or inactivated by other thiol-containing enzymes, cysteine, glutathione, etc.) And affecting the efficacy (see the following formula), its clinical dose reached 560mg / day, and the burden on patients is increased, so there is still a need to develop a more efficient BTK inhibitor for the treatment of related diseases.
  • the new molecule has a double reaction site, and after being in vivo (or intracellularly) by other thiol-containing enzymes, cysteine, glutathione, etc., it still has A site which reacts with a thiol group of a Btk enzyme cysteine (Cys481) residue, an alpha halogenated amide group (see the following formula), thereby exerts an activity enhancing effect.
  • the Bruton tyrosine kinase inhibitor provided by the present invention has the structure of Formula I:
  • the BTK inhibitor provided by the present invention has the following structure:
  • Rd 1, Rd 2, Rd 3, Rd 4, Rd 5 is independently H, halo, -CF 3, -CN, - NO 2 , -OH, C 1 -C 3 alkoxy group, or -NH 2
  • X is selected from fluorine, chlorine or bromine.
  • optical isomer thereof or a pharmaceutically acceptable salt or solvate thereof, wherein: X is selected from the group consisting of fluorine, chlorine or bromine.
  • the BTK inhibitor provided by the present invention has the structure of the general formula (IV):
  • Rd 1, Rd 2, Rd 3, Rd 4, Rd 5 is independently H, halo, -CF 3, -CN, - NO 2 , -OH, C 1 -C 3 alkoxy group, or -NH 2
  • X is selected from fluorine, chlorine or bromine
  • Y 1 , Y 2 are independently selected from C, N
  • Y 1 , Y 2 have at least One is selected from N.
  • optical isomer thereof or a pharmaceutically acceptable salt or solvate thereof, wherein: X is selected from fluorine, chlorine or bromine, Y 1 , Y 2 are independently selected from C, N, and Y 1 , Y 2 are at least One is selected from N.
  • aryl refers to an all-carbon monocyclic or fused polycyclic group of 5 to 12 carbon atoms having a fully conjugated pi-electron system.
  • aromatic rings are: benzene rings, naphthalene rings, and anthracene rings. The aromatic ring may be unsubstituted or substituted.
  • the substituent of the aromatic ring is selected from the group consisting of halogen, nitro, amino, C 1 -C 6 alkyl, C 1 -C 6 alkoxy, halogenated C 1 -C 6 alkyl, halogenated C 1 -C 6 alkoxy a C 3 -C 6 cycloalkyl group, a halogenated C 3 -C 6 cycloalkyl group;
  • heteroaryl refers to an unsaturated ring group of 5 to 12 ring atoms having a fully conjugated pi-electron system corresponding to one or more carbons of the above "aryl” being heteroatoms such as oxygen. Replacement with nitrogen, sulfur, etc.
  • the heteroaryl ring may be a single ring or a double ring, that is, fused by two rings.
  • Specific heterocyclic aryl (heteroaryl) groups may be: pyridyl, pyrimidinyl, pyrazinyl, isoxazolyl, isothiazolyl, pyrazolyl, thiazolyl, oxazolyl and imidazolyl, and the like.
  • the heterocyclic aryl group can be unsubstituted or substituted.
  • the substituent of the heterocyclic aryl group is selected from the group consisting of halogen, nitro, amino, C 1 -C 6 alkyl, C 1 -C 6 alkoxy, halogenated C 1 -C 6 alkyl, halogenated C 1 -C 6 Alkoxy, C 3 -C 6 cycloalkyl, halogenated C 3 -C 6 cycloalkyl;
  • alkoxy refers to an -O-alkyl group, wherein alkyl is as defined above.
  • alkoxy as used herein include, but are not limited to, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy and tert-butoxy.
  • Alkoxy also includes substituted alkoxy groups. The alkoxy group can be optionally substituted one or more times with a halogen.
  • the alkenyl moiety can be branched, straight or cyclic (in this case, it will also be referred to as "cycloalkenyl").
  • the alkenyl group may be a monovalent group or a divalent group (i.e., an alkenylene group).
  • the alkenyl group can be optionally substituted.
  • the alkenyl group may have 2 to 10 carbon atoms.
  • the alkenyl group can also be said to have a "lower alkenyl group" having 2 to 6 carbon atoms.
  • pharmaceutically acceptable derivative refers to a salt or solvate of a selected compound.
  • solvate refers to a variable stoichiometric complex formed from a solute (eg, a compound of the formula (I) to formula (V) of the present invention) and a solvent.
  • a solute eg, a compound of the formula (I) to formula (V) of the present invention
  • a solvent for the purposes of the present invention, the solvent does not interfere with the biological activity of the solute.
  • suitable solvents include, but are not limited to, water, methanol, ethanol, and acetic acid.
  • the solvent preferably used is a pharmaceutically acceptable solvent.
  • Suitable pharmaceutically acceptable solvents include, but are not limited to, water, ethanol, and acetic acid. More preferably, the solvent used is water.
  • Salts of the compounds of the invention can be prepared by the present invention using methods well known to those skilled in the art.
  • the salt may be an organic acid salt, a mineral acid salt or the like, and the organic acid salt includes a decanoate, a fumarate, an oxalate, a malate, a lactate, a camphor sulfonate, and a pair.
  • Tosylate, mesylate, etc.; the inorganic acid salt includes a hydrohalide, a sulfate, a phosphate, a nitrate, and the like.
  • a lower alkylsulfonic acid such as methanesulfonic acid, trifluoromethanesulfonic acid or the like may form a mesylate salt, a triflate salt; and an arylsulfonic acid such as benzenesulfonic acid or p-toluenesulfonic acid.
  • p-toluenesulfonate besylate; forming an appropriate salt with an organic carboxylic acid such as acetic acid, fumaric acid, tartaric acid, oxalic acid, maleic acid, malic acid, succinic acid or citric acid;
  • glutamic acid or aspartic acid can form glutamate or aspartate.
  • Corresponding salts may also be formed with inorganic acids such as hydrohalic acids (e.g., hydrofluoric acid, hydrobromic acid, hydroiodic acid, hydrochloric acid), nitric acid, carbonic acid, sulfuric acid or phosphoric acid.
  • hydrohalic acids e.g., hydrofluoric acid, hydrobromic acid, hydroiodic acid, hydrochloric acid
  • nitric acid e.g., carbonic acid, sulfuric acid or phosphoric acid.
  • a second object of the present invention is to provide a pharmaceutical composition comprising at least one active ingredient together with one or more pharmaceutically acceptable carriers or excipients, said active ingredient It may be a BTK inhibitor compound of the present invention, an optical isomer of the compound, the compound or an optical isomer thereof in a pharmaceutically acceptable salt, a solvate of the compound or an optical isomer thereof Any one or any of a variety.
  • the carrier includes conventional diluents, excipients, fillers, binders, wetting agents, disintegrating agents, absorption enhancers, surfactants, adsorption carriers, lubricants, etc. in the pharmaceutical field, and may also be added if necessary. Agent, sweetener, etc.
  • the medicament of the present invention can be prepared into various forms such as tablets, powders, granules, capsules, oral liquids and injectable preparations, and the medicaments of the above respective dosage forms can be prepared according to a conventional method in the pharmaceutical field.
  • the present invention provides the use of a compound of the formula (I) to formula (V) as disclosed herein, and an optical isomer thereof, or a pharmaceutically acceptable salt or solvate thereof thereof, to inhibit Bruce A tyrosine kinase (Btk) activity or a disease, disorder or condition that benefits from inhibition of Bruton's tyrosine kinase (Btk) activity.
  • composition comprising a therapeutically effective amount of at least one compound administered to a subject in need thereof, thereby inhibiting Bruton's tyrosine kinase activity of said subject.
  • a method wherein the compound has a structural formula of the formula (I) to the formula (V).
  • 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 (Still's disease), juvenile arthritis, diabetes, myasthenia gravis, Hashimoto's thyroiditis, Ord's thyroiditis, Graves'disease, rheumatoid arthritis Syndrome Syndrome), multiple sclerosis, Guillain-Barré syndrome, acute disseminated encephalomyelitis, Addison's disease, visual ocular palsy-myoclonus syndrome, coercion Spondylitis, antiphospholipid antibody syndrome, aplastic anemia, autoimmune hepatitis, coeliac disease, Goodpasture's syndrome, idiopathic thrombocytopenic purpura, optic neuritis, Scleroderma, primary biliary cirrhosis, Reiter's syndrome, Takayas
  • a subject in need thereof has cancer.
  • the cancer is a B cell proliferative disorder, such as diffuse large B-cell lymphoma, follicular lymphoma, chronic lymphocytic lymphoma, chronic lymphocytic leukemia, B-cell pro-lymphocytic leukemia, Lymphocyte plasma lymphoma/Waldenstrom macroglobulinemia Macroglobulinemia), spleen marginal lymphoma, plasma cell myeloma, plasmacytoma, extranodal marginal zone B-cell lymphoma, lymph node marginal zone B-cell lymphoma, mantle cell lymphoma, mediastinum (thymus) large B-cell lymph Tumor, intravascular large B-cell lymphoma, primary exudative lymphoma, Burkitt lymphoma/leukemia or lymphomatoid granulomatosis.
  • B cell proliferative disorder such as diffuse large B-cell lymphoma, follicular lymphom
  • the invention also provides the use of a compound of the invention or a pharmaceutically acceptable salt thereof for the preparation of a BTK inhibitor, in particular for the preparation of a medicament for the treatment of a cell proliferative disorder.
  • the cell proliferative diseases include cancer.
  • the present invention also provides a compound of the formula (I) to the formula (V), and an optical isomer thereof, or a pharmaceutically acceptable salt or solvate thereof, alone or in combination with other drugs for treating hyperplasia Applications in diseases such as cancer.
  • Antineoplastic agents which can be used in combination with a compound provided by the present invention or a pharmaceutically acceptable salt thereof include, but are not limited to, at least one of the following classes: mitotic inhibitors (e.g., vinblastine, vindesine, and vinorelbine); tubulin Decomposition inhibitors (such as Taxol); alkylating agents (such as cisplatin, carboplatin and cyclophosphamide); antimetabolites (such as 5-fluorouracil, tegafur, methotrexate, cytarabine and hydroxyl Urea); insertable antibiotics (such as arrhenone, mitomycin and bleomycin); enzymes (such as aspartate); topoisomerase inhibitors (such as etoricin and camptothecin); biological response regulation Agent (such as interferon).
  • mitotic inhibitors e.g., vinblastine, vindesine, and vinorelbine
  • tubulin Decomposition inhibitors such as Taxol
  • the present invention also provides a process for the preparation of the general formula (I') and a pharmaceutically acceptable derivative thereof, which is synthesized by the synthetic route shown in the following scheme:
  • Compound 1 (prepared by the method of WO2012158795) is refluxed with R 1 B(OH) 2 in the presence of potassium phosphate, a palladium catalyst and a suitable solvent or a mixed solvent such as dioxane/water.
  • the obtained compound 2 is reacted with Boc-protected 3-hydroxypiperidine in the presence of triphenylphosphine, DIAD and a suitable solvent such as THF to give compound 3, followed by hydrolysis under acidic conditions to give key intermediate 4.
  • the key intermediate DCC and a suitable solvent such as DCM are condensed with a substituted acrylic acid moiety to give a compound of the formula (I').
  • R 1 corresponds to the compound of the above formula (I) to formula (V), and an appropriate substituted boronic acid can be selected according to actual needs.
  • substituted boronic acids include, but are not limited to, 4-phenoxybenzeneboronic acid, 4-(4-methoxyphenoxy)benzeneboronic acid, 4-(3-methoxyphenoxy)benzeneboronic acid, 4-( 3,4-Dimethoxyphenoxy)benzeneboronic acid, 4-(4-fluorophenoxy)benzeneboronic acid, 4-(4-chlorophenoxy)benzeneboronic acid, 6-phenoxypyridine-3- Boric acid, 6-(2-fluorophenoxy)pyridin-3-ylboronic acid, 6-(4-fluorophenoxy)pyridin-3-ylboronic acid and the like.
  • the present inventors have confirmed by experiments that the compound of the present invention has an anti-proliferative inhibitory effect on tumor cell lines such as A549, SGC7901, MCF-7, PC-9, HL-60, and the like, and can be applied to the treatment of human or animal cell proliferation-related entities.
  • tumor cell lines such as A549, SGC7901, MCF-7, PC-9, HL-60, and the like.
  • the inventors of the present invention have confirmed by experiments that the compound of the present invention has good pharmacokinetic properties and can be applied to orally treating solid tumors or blood cancers associated with cell proliferation of human or animal cells or suffering from autoimmune diseases.
  • the inventors of the present invention confirmed by experiments that the compound of the present invention has a two-site reaction property.
  • Example 41 In vitro Btk kinase inhibitory activity and in vitro antitumor activity assay
  • the drug was dissolved in DMSO to make a 10 mM stock solution, and diluted to a 50x test concentration.
  • the test concentration was diluted by 3 to 25nM, 8.33nM, 2.78nM, 0.93nM, 0.31nM, 0.10nM. .
  • 10 ⁇ L of 50x drug stock solution was added to a 96-well plate, and 90 ⁇ L of 1x kinase buffer was added and shaken on the shaker for 10 minutes. Transfer 5 ⁇ L from each well of a 96-well plate to a 384-well plate, and set 2 duplicate wells in a 384-well plate.
  • Kinase Reaction Prepare 2.5x Kinase Buffer: Add the enzyme to the 1x Kinase Buffer.
  • the antitumor activity of the synthesized compounds was determined by using different solid tumors and leukemia cell lines:
  • Cell lines human lung cancer cells (A549, PC9), human breast cancer cells (MCF7), human gastric cancer cells (SGC7901), acute promyelocytic leukemia cells (HL60), human lung cancer cells gefitinib-resistant cells (PC9) -IR).
  • MCF7 DMEM + fetal bovine serum
  • HL60 RPMI 1640+ fetal calf serum
  • PC9 DMEM + fetal bovine serum
  • PC9-IR DMEM + fetal bovine serum
  • Drug preparation method The drug was dissolved in DMSO to make a 10 mM stock solution, and diluted to a certain ratio to obtain 5 different concentrations (test concentration 100x).
  • the selected six tumor cells A549, MCF7, SGC7901, HL60, PC9, PC9-IR were incubated in a 37 ° C, 5% CO 2 cell incubator, and were passaged when the cell density was 70-90%. Cells were passaged with Duck's EDTA and passaged for later experiments.
  • Tumor cells A549, MCF7, SGC7901, HL60, PC9, PC9-IR were seeded in a 96-well plate at 4000 cells/200 ⁇ L/well and incubated overnight at 37 ° C in a 5% CO 2 cell incubator.
  • 2 ⁇ L of compound was added to each well to a final concentration of 50 ⁇ M, 10 ⁇ M, 2 ⁇ M, 0.4 ⁇ M, and 0.08 ⁇ M for 72 hours at 37 ° C in a 5% CO 2 cell incubator with DMSO (2%) as a control.
  • 20 ⁇ L of CCK-8 solution was added and placed in a 37 ° C, 5% CO 2 cell incubator for 4 hours.
  • cell inhibition rate % [(control group OD value - blank group OD value) - (medical group OD value - blank group OD value)] / (control cell OD value - blank group OD value) ⁇ 100%, the half-inhibitory concentration (IC 50 ) was calculated by CalcuSyn software.
  • the pharmacokinetic properties of compounds 5-2, 5-3 and 5-19 in rats were investigated.
  • the specific methods were as follows: SD rats were used as experimental animals, and given by gavage The drug was administered at a dose of 20 mg/kg, and 5 mg/kg was administered intravenously to the tail vein.
  • the time of blood collection from the tail vein of the intragastric administration was 0.17, 0.33, 0.5, 1, 1.5, 2, 4, 6, 8, 12, 24 hours; the time of blood collection by intravenous administration was 0.05, 0.1, 0.17, 0.5. 1,2,4,6,8,12,24 hours.
  • 0.3 ml of whole blood was taken, and 0.1 ml of plasma was taken after centrifugation and analyzed by LC-MS.
  • Example 45 Treatment of differential arthritis with compounds 5-2 and 5-19

Abstract

The present invention provides a dual-site irreversible Bruton's tyrosine kinase inhibitor, a composition and application therefof. The composition of the present invention has an anti-proliferation effect on tumor cell strains such as A549, SGC7901, MCF-7, HL-60 and PC9-IR, and can be applied to a medicine for treating solid tumors or blood cancer related to human or animal cell proliferation; the composition of the present invention has better pharmacokinetic property, and can be applied to oral administration to treat the solid tumors, the blood cancer or autoimmune diseases related to the human or animal cell proliferation; and the composition of the present invention has the characteristic of dual-site reaction.

Description

双位点不可逆布鲁顿酪氨酸激酶抑制剂、组合物及其应用Two-site irreversible Bruton's tyrosine kinase inhibitor, composition and application thereof 技术领域Technical field
本发明属于医药领域，具体是一种双位点不可逆布鲁顿酪氨酸激酶抑制剂、组合物及其应用。The invention belongs to the field of medicine, in particular to a two-site irreversible Bruton tyrosine kinase inhibitor, a composition and application thereof.
背景技术Background technique
小分子共价抑制剂(covalent inhibitors)，也称不可逆抑制剂(irreversible inhibitors)，是通过共价键与靶蛋白残基发生不可逆结合，从而发挥其生物学功能的一类抑制剂。共价抑制剂药物在过去的几十年里对人类健康做出了重要贡献。相对于非共价抑制剂，共价抑制剂通过与靶蛋白以共价键结合增强了与靶标的亲和性，这是共价抑制剂表现其高生物活性的根本原因。近年来,由于非共价靶向抗肿瘤药物特别是大量针对激酶的替尼类药物耐药的产生，使人们又更多地关注共价抑制剂药物。近年来，许多大型制药公司均开展了针对特定酶靶点的共价抑制剂的研发，目前已有部分共价抑制剂进入临床试验，包括阿法替尼、卡那替尼、来那替尼等。其中，阿法替尼已于2013年7月12日被美国FDA正式批准用于治疗表皮生长因子受体(EGFR)基因突变的转移性非小细胞肺癌，成为首个被FDA批准的治疗肺癌的不可逆抑制剂新药。此外，抗病毒的共价药物也是近年来的研究热点，并且已取得了很大的进展，例如，2011年FDA已批准了两个抗丙型肝炎病毒共价抑制剂药物，即特拉匹韦(Telaprevir)和波普瑞韦(Boceprevir)。这些研究证明了不可逆抑制剂可有效用于疾病的治疗。Small molecule covalent inhibitors, also known as irreversible inhibitors, are a class of inhibitors that exert their biological functions by irreversible binding of covalent bonds to target protein residues. Covalent inhibitor drugs have made important contributions to human health over the past few decades. Covalent inhibitors enhance affinity to the target by covalent bonding to the target protein relative to the non-covalent inhibitor, which is the underlying cause of the high bioactivity of the covalent inhibitor. In recent years, due to the non-covalent targeting of anti-tumor drugs, especially the production of a large number of tini-resistant drugs against kinases, people have paid more attention to covalent inhibitor drugs. In recent years, many large pharmaceutical companies have developed covalent inhibitors for specific enzyme targets. Currently, some covalent inhibitors have entered clinical trials, including afatinib, neratinib, and natenitny. Wait. Among them, afatinib was officially approved by the US FDA on July 12, 2013 for the treatment of metastatic non-small cell lung cancer with epidermal growth factor receptor (EGFR) gene mutation, becoming the first FDA-approved treatment of lung cancer. New drug for irreversible inhibitors. In addition, antiviral covalent drugs have also been a research hotspot in recent years, and great progress has been made. For example, in 2011, the FDA approved two anti-hepatitis C virus covalent inhibitor drugs, namely, telaprevir. (Telaprevir) and Boceprevir (Boceprevir). These studies demonstrate that irreversible inhibitors are effective for the treatment of disease.
布鲁顿酪氨酸激酶(Bruton's tyrosine kinase,Btk)，一种非受体酪氨酸激酶Tec家族的成员，是在除了T淋巴细胞和自然杀伤细胞之外的所有造血细胞类型中表达的关键信号酶。Btk在连接细胞表面B细胞受体(B-cell receptor，BCR)刺激至下游细胞内应答的B细胞信号传导途径中扮演至关重要的角色。Btk是B细胞发育、激活、信号传导和存活的关键调节物。另外，Bkt在众多其他造血细胞信号传导途径中起作用，例如在巨噬细胞中的Toll样受体(Toll like receptor，TLR)和细胞因子受体介导的TNF-α产生、在肥大细胞中的免疫球蛋白E受体(FcεR1)信号传导、在B-谱系淋巴样细胞中抑制Fas/APO-1细胞凋亡的信号传导以及胶原刺激的血小板聚集。参见例如C.A.Jeffries等，J.Bio.Chem.(2003)278:26258-26264、N.J.Horwood等，J.Exp.Med.(2003)197:1603-1611。近年来研究显示，Btk信号通路是目前非霍奇金淋巴瘤(NHL)，特别是慢性淋巴细胞白血病(CLL)、B细胞淋巴瘤及自身免疫疾病临床治疗研究中的新热点。小分子Btk抑制剂通过作用于BCR信号通路，与Btk结合而抑制Btk自身磷酸化，阻止Btk的激活，从而阻断细胞传导并诱导细胞凋亡。Btk抑制剂选择性强，毒副作用低，特别是伊布替尼的上市，被FDA定为“突破性”新药，其研究开发前景广阔。伊布替尼与Btk酶半胱氨酸(Cys481)残基的巯基发生反应，并形成共价键，使Btk酶失活而发挥疗效。然而，伊布替尼替尼在给药过程中，易被代谢(被代谢酶氧化代谢成双羟化产物或者被其他含巯基的酶、半胱氨酸、谷胱甘肽等进攻而失活)而影响药效(见下式)，其临床给药剂量达到了560mg/天，而使病人负担加重，因此仍需发展一类更为高效的BTK抑制剂用于相关疾病的治疗。 Bruton's tyrosine kinase (Btk), a member of the Tec family of non-receptor tyrosine kinases, is key for expression in all hematopoietic cell types except T lymphocytes and natural killer cells. Signal enzyme. Btk plays a crucial role in the B cell signaling pathway that links cell surface B-cell receptor (BCR) to downstream cell responses. Btk is a key regulator of B cell development, activation, signaling, and survival. In addition, Bkt plays a role in numerous other hematopoietic signaling pathways, such as Toll like receptor (TLR) and cytokine receptor-mediated TNF-α production in macrophages, in mast cells. Immunoglobulin E receptor (FcεR1) signaling, signaling that inhibits apoptosis of Fas/APO-1 cells in B-lineage lymphoid cells, and collagen-stimulated platelet aggregation. See, for example, C. A. Jeffries et al, J. Bio. Chem. (2003) 278: 26258-26264, N. J. Horwood et al, J. Exp. Med. (2003) 197: 1603-1611. Recent studies have shown that the Btk signaling pathway is a new hotspot in the current clinical research of non-Hodgkin's lymphoma (NHL), especially chronic lymphocytic leukemia (CLL), B-cell lymphoma and autoimmune diseases. Small molecule Btk inhibitors inhibit Btk autophosphorylation by binding to Btk by acting on the BCR signaling pathway, preventing Btk activation, thereby blocking cell conduction and inducing apoptosis. Btk inhibitors have strong selectivity and low toxic and side effects, especially the listing of ibufenib, which has been designated as a “breakthrough” new drug by the FDA, and its research and development prospects are broad. Ibbutinib reacts with the sulfhydryl group of the Btk enzyme cysteine (Cys481) residue and forms a covalent bond, which inactivates the Btk enzyme and exerts its therapeutic effect. However, ibbutinibinib is easily metabolized during metabolism (digested by metabolic enzymes to be dihydroxylated or inactivated by other thiol-containing enzymes, cysteine, glutathione, etc.) And affecting the efficacy (see the following formula), its clinical dose reached 560mg / day, and the burden on patients is increased, so there is still a need to develop a more efficient BTK inhibitor for the treatment of related diseases.
Figure PCTCN2016081669-appb-000001
Figure PCTCN2016081669-appb-000001
发明内容Summary of the invention
本发明的目的在于提供新颖的、未见文献报道的双位点BTK不可逆抑制剂或其光学异构体或其药学上可接受的盐或溶剂合物。基于伊布替尼的体内代谢途径分析，假如在伊布替尼的α,β-不饱和双键的近端引入卤素原子，使得其代谢位点被占据而阻断双键被代谢酶所双羟化，进而提高其体内的代谢稳定性。更为重要的是，由于卤素原子的引入，使得新分子具有了双反应位点，在体内(或者细胞内)被其他含巯基的酶、半胱氨酸、谷胱甘肽等后，仍然具有一个可与Btk酶半胱氨酸(Cys481)残基的巯基发生反应的位点---α卤代的酰胺基团(见下式)，进而起到活性增强的效果。It is an object of the present invention to provide novel, non-reported two-site BTK irreversible inhibitors or optical isomers thereof, or pharmaceutically acceptable salts or solvates thereof. Based on the analysis of the in vivo metabolic pathway of Ibubinib, if the halogen atom is introduced at the proximal end of the α,β-unsaturated double bond of Ibubinib, the metabolic site is occupied and the double bond is blocked by the metabolic enzyme. Hydroxylation, which in turn increases metabolic stability in the body. More importantly, due to the introduction of halogen atoms, the new molecule has a double reaction site, and after being in vivo (or intracellularly) by other thiol-containing enzymes, cysteine, glutathione, etc., it still has A site which reacts with a thiol group of a Btk enzyme cysteine (Cys481) residue, an alpha halogenated amide group (see the following formula), thereby exerts an activity enhancing effect.
Figure PCTCN2016081669-appb-000002
Figure PCTCN2016081669-appb-000002
由于伊布替尼在水中的溶解度较低，在其芳香环中引入氮原子，可以提高其水溶解性。 Since the solubility of ibbutinib in water is low, the introduction of nitrogen atoms into its aromatic ring can improve its water solubility.
本发明采用如下的技术方案：The invention adopts the following technical solutions:
本发明所提供的布鲁顿酪氨酸激酶抑制剂，其具有通式Ⅰ的结构：The Bruton tyrosine kinase inhibitor provided by the present invention has the structure of Formula I:
Figure PCTCN2016081669-appb-000003
Figure PCTCN2016081669-appb-000003
或其光学异构体或其药学上可接受的盐或溶剂合物，其中：Ra,Rb,Rc独立选自H、卤素、-CF3、-CN、-NO2、OH、NH2、-L-C1-C6的烷基、-L-C1-C6的烯基、-L-取代或非取代的杂芳基、或-L-取代或非取代的芳基，其中L是键、O、S、-S(＝O)、-S(＝O)2、NH、C(O)、CH2、-NHC(O)O、-NHC(O)或-C(O)NH，X选自氟、氯或溴，Y1,Y2独立选自C、N。Or an optical isomer thereof, or a pharmaceutically acceptable salt or solvate thereof, wherein: Ra, Rb, Rc are independently selected from the group consisting of H, halogen, -CF 3 , -CN, -NO 2 , OH, NH 2 , - LC alkyl, -LC 1 -C 6 alkenyl group of the 1 -C 6, -L- aryl substituted or unsubstituted heteroaryl group, or -L-substituted or unsubstituted aryl group, wherein L is a bond, O, S, -S(=O), -S(=O) 2 , NH, C(O), CH 2 , -NHC(O)O, -NHC(O) or -C(O)NH, X is selected from Fluorine, chlorine or bromine, Y 1 , Y 2 are independently selected from C and N.
本发明所提供的BTK抑制剂具有如下结构：The BTK inhibitor provided by the present invention has the following structure:
Figure PCTCN2016081669-appb-000004
Figure PCTCN2016081669-appb-000004
或其光学异构体或其药学上可接受的盐或溶剂合物，其中：Ra,Rb,Rc独立选自H、卤素、-CF3、-CN、-NO2、OH、NH2、-L-C1-C6的烷基、-L-C1-C6的烯基、-L-取代或非取代的杂芳基、或-L-取代或非取代的芳基，其中L是键、O、S、-S(＝O)、-S(＝O)2、NH、C(O)、CH2、-NHC(O)O、-NHC(O)或-C(O)NH，X选自氟、氯或溴。Or an optical isomer thereof, or a pharmaceutically acceptable salt or solvate thereof, wherein: Ra, Rb, Rc are independently selected from the group consisting of H, halogen, -CF 3 , -CN, -NO 2 , OH, NH 2 , - LC alkyl, -LC 1 -C 6 alkenyl group of the 1 -C 6, -L- aryl substituted or unsubstituted heteroaryl group, or -L-substituted or unsubstituted aryl group, wherein L is a bond, O, S, -S(=O), -S(=O) 2 , NH, C(O), CH 2 , -NHC(O)O, -NHC(O) or -C(O)NH, X is selected from Fluorine, chlorine or bromine.
更进一步地，本发明优选的化合物具有通式(II)结构：Still further, preferred compounds of the invention have the structure of formula (II):
Figure PCTCN2016081669-appb-000005
Figure PCTCN2016081669-appb-000005
或其光学异构体或其药学上可接受的盐或溶剂合物，其中：Rd1、Rd2、Rd3、Rd4、Rd5独立地是H、卤素、-CF3、-CN、-NO2、-OH、C1-C3的烷氧基、或-NH2，X选自氟、氯或溴。 Or an optical isomer or a pharmaceutically acceptable salt or solvate thereof, wherein: Rd 1, Rd 2, Rd 3, Rd 4, Rd 5 is independently H, halo, -CF 3, -CN, - NO 2 , -OH, C 1 -C 3 alkoxy group, or -NH 2 , X is selected from fluorine, chlorine or bromine.
更进一步地，本发明优选的化合物具有通式(III)结构：Still further, preferred compounds of the invention have the structure of formula (III):
Figure PCTCN2016081669-appb-000006
Figure PCTCN2016081669-appb-000006
或其光学异构体或其药学上可接受的盐或溶剂合物，其中：X选自氟、氯或溴。Or an optical isomer thereof, or a pharmaceutically acceptable salt or solvate thereof, wherein: X is selected from the group consisting of fluorine, chlorine or bromine.
本发明所提供的BTK抑制剂具有通式(IV)结构：The BTK inhibitor provided by the present invention has the structure of the general formula (IV):
Figure PCTCN2016081669-appb-000007
Figure PCTCN2016081669-appb-000007
或其光学异构体或其药学上可接受的盐或溶剂合物，其中：Rd1、Rd2、Rd3、Rd4、Rd5独立地是H、卤素、-CF3、-CN、-NO2、-OH、C1-C3的烷氧基、或-NH2，X选自氟、氯或溴,Y1,Y2独立选自C、N,且Y1,Y2至少有一个选自N。Or an optical isomer or a pharmaceutically acceptable salt or solvate thereof, wherein: Rd 1, Rd 2, Rd 3, Rd 4, Rd 5 is independently H, halo, -CF 3, -CN, - NO 2 , -OH, C 1 -C 3 alkoxy group, or -NH 2 , X is selected from fluorine, chlorine or bromine, Y 1 , Y 2 are independently selected from C, N, and Y 1 , Y 2 have at least One is selected from N.
更进一步地，本发明优选的化合物具有通式(V)结构：Still further, preferred compounds of the invention have the structure of formula (V):
Figure PCTCN2016081669-appb-000008
Figure PCTCN2016081669-appb-000008
或其光学异构体或其药学上可接受的盐或溶剂合物，其中：X选自氟、氯或溴,Y1,Y2独立选自C、N,且Y1,Y2至少有一个选自N。Or an optical isomer thereof, or a pharmaceutically acceptable salt or solvate thereof, wherein: X is selected from fluorine, chlorine or bromine, Y 1 , Y 2 are independently selected from C, N, and Y 1 , Y 2 are at least One is selected from N.
术语说明：本文所用术语“芳基”是指5到12个碳原子的全碳单环或稠合多环基团，具有完全共轭的π电子***。芳环的非限制性实例有：苯环、萘环和蒽环。芳环可以是无取代或取代的。芳环的取代基选自卤素、硝基、氨基、C1-C6烷基、C1-C6烷氧基、卤代C1-C6烷基、卤代C1-C6烷氧基、C3-C6环烷基、卤代C3-C6环烷基；Description of terms: As used herein, the term "aryl" refers to an all-carbon monocyclic or fused polycyclic group of 5 to 12 carbon atoms having a fully conjugated pi-electron system. Non-limiting examples of aromatic rings are: benzene rings, naphthalene rings, and anthracene rings. The aromatic ring may be unsubstituted or substituted. The substituent of the aromatic ring is selected from the group consisting of halogen, nitro, amino, C 1 -C 6 alkyl, C 1 -C 6 alkoxy, halogenated C 1 -C 6 alkyl, halogenated C 1 -C 6 alkoxy a C 3 -C 6 cycloalkyl group, a halogenated C 3 -C 6 cycloalkyl group;
本文所用术语“杂芳基”指5到12个环原子的不饱和的环基团，具有完全共轭的π电子系 统，相当于上述“芳基”中一个或多个碳被杂原子例如氧、氮、硫等置换。杂芳环可以是单环，也可以是双环，即通过两个环稠合而成。具体的杂环芳基(杂芳基)可以是：吡啶基，嘧啶基，吡嗪基，异恶唑基，异噻唑基、吡唑基、噻唑基、恶唑基和咪唑基等。杂环芳基可以是无取代或取代的。杂环芳基的取代基选自卤素、硝基、氨基、C1-C6烷基、C1-C6烷氧基、卤代C1-C6烷基、卤代C1-C6烷氧基、C3-C6环烷基、卤代C3-C6环烷基；The term "heteroaryl" as used herein, refers to an unsaturated ring group of 5 to 12 ring atoms having a fully conjugated pi-electron system corresponding to one or more carbons of the above "aryl" being heteroatoms such as oxygen. Replacement with nitrogen, sulfur, etc. The heteroaryl ring may be a single ring or a double ring, that is, fused by two rings. Specific heterocyclic aryl (heteroaryl) groups may be: pyridyl, pyrimidinyl, pyrazinyl, isoxazolyl, isothiazolyl, pyrazolyl, thiazolyl, oxazolyl and imidazolyl, and the like. The heterocyclic aryl group can be unsubstituted or substituted. The substituent of the heterocyclic aryl group is selected from the group consisting of halogen, nitro, amino, C 1 -C 6 alkyl, C 1 -C 6 alkoxy, halogenated C 1 -C 6 alkyl, halogenated C 1 -C 6 Alkoxy, C 3 -C 6 cycloalkyl, halogenated C 3 -C 6 cycloalkyl;
本文所用术语“烷氧基”是指-O-烷基基团，其中烷基如上所定义。本文所用“烷氧基”的实例包括但不限于甲氧基、乙氧基、正丙氧基、异丙氧基、正丁氧基和叔丁氧基。“烷氧基”还包括取代烷氧基。烷氧基可任选被卤素取代一次或多次。The term "alkoxy" as used herein, refers to an -O-alkyl group, wherein alkyl is as defined above. Examples of "alkoxy" as used herein include, but are not limited to, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy and tert-butoxy. "Alkoxy" also includes substituted alkoxy groups. The alkoxy group can be optionally substituted one or more times with a halogen.
术语“烯基”是指一类烷基，其中烷基的起始的两个原子形成双键，该双键不是芳香基的组成部分。也就是说，烯基始于原子-C(R1)＝C(R2)-R3，其中R1、R2、R3是指烯基的其余部分，R1、R2、R3可以相同或不同。烯基部分可以是支链、直链或环状的(在此情况下，它也会被称为“环烯基”)。根据结构，烯基可以是单价基团或双价基团(即亚烯基)。烯基可以是任选取代的。烯基的非限制性实例包括-CH＝CH2、-C(CH3)＝CH2、-CH＝CHCH3、-C(CH3)＝CHCH3。亚烯基包括但不限于-CH＝CH-、-C(CH3)＝CH-、-CH＝CHCH2-、-CH＝CHCH2CH2-和-C(CH3)＝CHCH2-。烯基可以具有2-10个碳原子。烯基也可以说具有2-6个碳原子的“低级烯基”。The term "alkenyl" refers to a class of alkyl groups wherein the first two atoms of the alkyl group form a double bond which is not part of the aryl group. That is, the alkenyl group begins with the atom -C(R 1 )=C(R 2 )-R 3 , wherein R 1 , R 2 , R 3 refer to the remainder of the alkenyl group, R 1 , R 2 , R 3 Can be the same or different. The alkenyl moiety can be branched, straight or cyclic (in this case, it will also be referred to as "cycloalkenyl"). Depending on the structure, the alkenyl group may be a monovalent group or a divalent group (i.e., an alkenylene group). The alkenyl group can be optionally substituted. Non-limiting examples of alkenyl groups include -CH=CH 2 , -C(CH 3 )=CH 2 , -CH=CHCH 3 , -C(CH 3 )=CHCH 3 . Alkenylene groups include, but are not limited to -CH = CH -, - C ( CH 3) = CH -, - CH = CHCH 2 -, - CH = CHCH 2 CH 2 - and -C (CH 3) = CHCH 2 -. The alkenyl group may have 2 to 10 carbon atoms. The alkenyl group can also be said to have a "lower alkenyl group" having 2 to 6 carbon atoms.
术语“药学可接受衍生物”是指所选化合物的盐或溶剂合物。The term "pharmaceutically acceptable derivative" refers to a salt or solvate of a selected compound.
本文所用术语“溶剂合物”是指由溶质(例如：本发明的通式(I)～通式(V)化合物)和溶剂形成的可变化学计量的复合物。为了本发明的目的，所述溶剂不能干扰溶质的生物学活性。合适的溶剂的实例包括但不限于水、甲醇、乙醇和乙酸。优选使用的溶剂为药学可接受溶剂。合适的药学可接受溶剂包括但不限于水、乙醇和乙酸。更优选地，所用溶剂为水。The term "solvate" as used herein refers to a variable stoichiometric complex formed from a solute (eg, a compound of the formula (I) to formula (V) of the present invention) and a solvent. For the purposes of the present invention, the solvent does not interfere with the biological activity of the solute. Examples of suitable solvents include, but are not limited to, water, methanol, ethanol, and acetic acid. The solvent preferably used is a pharmaceutically acceptable solvent. Suitable pharmaceutically acceptable solvents include, but are not limited to, water, ethanol, and acetic acid. More preferably, the solvent used is water.
本发明采用本领域技术人员所熟知的方法可以制备本发明所述化合物的盐。所述的盐可以是有机酸盐、无机酸盐等，所述的有机酸盐包括枸橼酸盐、富马酸盐、草酸盐、苹果酸盐、乳酸盐、樟脑磺酸盐、对甲苯磺酸盐、甲磺酸盐等；所述的无机酸盐包括氢卤酸盐、硫酸盐、磷酸盐、硝酸盐等。例如，与低级烷基磺酸，如甲磺酸，三氟甲磺酸等可形成甲磺酸盐、三氟甲磺酸盐；与芳基磺酸，如苯磺酸或对甲苯磺酸等可形成对甲苯磺酸盐、苯磺酸盐；与有机羧酸，如乙酸，富马酸，酒石酸，草酸，马来酸，苹果酸，琥珀酸或柠檬酸等可形成相应的盐；与氨基酸，如谷氨酸或天冬氨酸可形成谷氨酸盐或天冬氨酸盐。与无机酸，如氢卤酸(如氢氟酸、氢溴酸、氢碘酸、氢氯酸)，硝酸，碳酸，硫酸或磷酸等也可形成相应的盐。Salts of the compounds of the invention can be prepared by the present invention using methods well known to those skilled in the art. The salt may be an organic acid salt, a mineral acid salt or the like, and the organic acid salt includes a decanoate, a fumarate, an oxalate, a malate, a lactate, a camphor sulfonate, and a pair. Tosylate, mesylate, etc.; the inorganic acid salt includes a hydrohalide, a sulfate, a phosphate, a nitrate, and the like. For example, a lower alkylsulfonic acid such as methanesulfonic acid, trifluoromethanesulfonic acid or the like may form a mesylate salt, a triflate salt; and an arylsulfonic acid such as benzenesulfonic acid or p-toluenesulfonic acid. Forming p-toluenesulfonate, besylate; forming an appropriate salt with an organic carboxylic acid such as acetic acid, fumaric acid, tartaric acid, oxalic acid, maleic acid, malic acid, succinic acid or citric acid; For example, glutamic acid or aspartic acid can form glutamate or aspartate. Corresponding salts may also be formed with inorganic acids such as hydrohalic acids (e.g., hydrofluoric acid, hydrobromic acid, hydroiodic acid, hydrochloric acid), nitric acid, carbonic acid, sulfuric acid or phosphoric acid.
本发明的第二个目的是提供一种药物组合物，所述药物组合物包含至少一种活性组分以及一种或多种药学上可接受的载体或赋形剂，所述的活性组分可以是本发明的BTK抑制剂化合物、所述化合物的光学异构体、所述化合物或其光学异构体在药学上可接受的盐、所述化合物或其光学异构体的溶剂合物中的任意一种或任意多种。A second object of the present invention is to provide a pharmaceutical composition comprising at least one active ingredient together with one or more pharmaceutically acceptable carriers or excipients, said active ingredient It may be a BTK inhibitor compound of the present invention, an optical isomer of the compound, the compound or an optical isomer thereof in a pharmaceutically acceptable salt, a solvate of the compound or an optical isomer thereof Any one or any of a variety.
所述载体包括药学领域的常规稀释剂，赋形剂，填充剂，粘合剂，湿润剂，崩解剂，吸收促进剂，表面活性剂，吸附载体，润滑剂等，必要时还可以加入香味剂，甜味剂等。本发明药物可以制成片剂，粉剂，粒剂，胶囊，口服液及注射用药等多种形式，上述各剂型的药物均可以按照药学领域的常规方法制备。The carrier includes conventional diluents, excipients, fillers, binders, wetting agents, disintegrating agents, absorption enhancers, surfactants, adsorption carriers, lubricants, etc. in the pharmaceutical field, and may also be added if necessary. Agent, sweetener, etc. The medicament of the present invention can be prepared into various forms such as tablets, powders, granules, capsules, oral liquids and injectable preparations, and the medicaments of the above respective dosage forms can be prepared according to a conventional method in the pharmaceutical field.
另一方面，本发明提供的是使用本文公开的通式(I)～通式(V)所述的化合物及其光学异构体或其药学上可接受的盐或溶剂合物来抑制布鲁顿酪氨酸激酶(Btk)活性或者治疗从布鲁顿酪氨酸激酶(Btk)活性的抑制中获益的疾病、障碍或病症。In another aspect, the present invention provides the use of a compound of the formula (I) to formula (V) as disclosed herein, and an optical isomer thereof, or a pharmaceutically acceptable salt or solvate thereof thereof, to inhibit Bruce A tyrosine kinase (Btk) activity or a disease, disorder or condition that benefits from inhibition of Bruton's tyrosine kinase (Btk) activity.
在进一步优选的方案中，本文提供的是通过给予有需要的治疗者一种含有治疗有效量的至 少一种化合物的组合物、从而抑制所述受治疗者的布鲁顿酪氨酸激酶活性的方法，其中所述化合物的结构式为通式(I)～通式(V)。在一些实施方式中，有需要的受治疗者罹患自身免疫性疾病，例如炎性肠病、关节炎、狼疮、类风湿性关节炎、银屑病性关节炎、骨关节炎、斯蒂尔病(Still’s disease)、青少年关节炎、糖尿病、重症肌无力症、桥本甲状腺炎(Hashimoto’s thyroiditis)、奥德甲状腺炎(Ord’s thyroiditis)、格雷夫斯氏病(Graves’disease)、类风湿性关节炎综合征(
Figure PCTCN2016081669-appb-000009
syndrome)、多发性硬化症、传染性神经元炎(Guillain-Barré syndrome)、急性播散性脑脊髓炎、阿狄森病(Addison’s disease)、视性眼阵挛-肌阵挛综合征、强制性脊椎炎、抗磷脂抗体综合征、再生障碍性贫血、自身免疫性肝炎、乳糜泻(coeliac disease)、古德帕斯彻综合征(Goodpasture’s syndrome)、特发性血小板减少性紫癜、视神经炎、硬皮病、原发性胆汁性肝硬化、莱特尔综合征(Reiter’s syndrome)、高安动脉炎(Takayasu’s arteritis)、颞动脉炎、温型自身免疫性溶血性贫血、韦格纳肉芽肿病(Wegener’s granulomatosis)、银屑病、全身脱毛、贝赫切特病(
Figure PCTCN2016081669-appb-000010
disease)、慢性疲劳、家族性自主神经功能异常、子宫内膜异位、间质性膀胱炎、神经肌强直、硬皮病或外阴痛。In a further preferred embodiment, provided herein is a composition comprising a therapeutically effective amount of at least one compound administered to a subject in need thereof, thereby inhibiting Bruton's tyrosine kinase activity of said subject. A method wherein the compound has a structural formula of the formula (I) to the formula (V). 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 (Still's disease), juvenile arthritis, diabetes, myasthenia gravis, Hashimoto's thyroiditis, Ord's thyroiditis, Graves'disease, rheumatoid arthritis Syndrome
Figure PCTCN2016081669-appb-000009
Syndrome), multiple sclerosis, Guillain-Barré syndrome, acute disseminated encephalomyelitis, Addison's disease, visual ocular palsy-myoclonus syndrome, coercion Spondylitis, antiphospholipid antibody syndrome, aplastic anemia, autoimmune hepatitis, coeliac disease, Goodpasture's syndrome, idiopathic thrombocytopenic purpura, optic neuritis, Scleroderma, primary biliary cirrhosis, Reiter's syndrome, Takayasu's arteritis, temporal arteritis, warm autoimmune hemolytic anemia, Wegener's granulomatosis (Wegener's Granulomatosis), psoriasis, generalized hair loss, Behcet's disease (
Figure PCTCN2016081669-appb-000010
Disease), chronic fatigue, familial autonomic dysfunction, endometriosis, interstitial cystitis, neuromuscular rigidity, scleroderma or vulvar pain.
在进一步的实施方式中，有需要的受治疗者罹患癌症。在一个实施方式中，所述癌症是B细胞增生性疾病，例如弥漫性大B细胞淋巴瘤、滤泡性淋巴瘤、慢性淋巴细胞淋巴瘤、慢性淋巴细胞白血病、B细胞前淋巴细胞性白血病、淋巴浆细胞淋巴瘤/瓦尔登斯特伦巨球蛋白血症(
Figure PCTCN2016081669-appb-000011
macroglobulinemia)、脾边缘区淋巴瘤、浆细胞性骨髓瘤、浆细胞瘤、结外边缘区B细胞淋巴瘤、***边缘区B细胞淋巴瘤、外套细胞淋巴瘤、纵膈(胸腺)大B细胞淋巴瘤、血管内大B细胞淋巴瘤、原发性渗出性淋巴瘤、伯基特淋巴瘤(Burkitt lymphoma)/白血病或淋巴瘤样肉芽肿病。In a further embodiment, a subject in need thereof has cancer. In one embodiment, the cancer is a B cell proliferative disorder, such as diffuse large B-cell lymphoma, follicular lymphoma, chronic lymphocytic lymphoma, chronic lymphocytic leukemia, B-cell pro-lymphocytic leukemia, Lymphocyte plasma lymphoma/Waldenstrom macroglobulinemia
Figure PCTCN2016081669-appb-000011
Macroglobulinemia), spleen marginal lymphoma, plasma cell myeloma, plasmacytoma, extranodal marginal zone B-cell lymphoma, lymph node marginal zone B-cell lymphoma, mantle cell lymphoma, mediastinum (thymus) large B-cell lymph Tumor, intravascular large B-cell lymphoma, primary exudative lymphoma, Burkitt lymphoma/leukemia or lymphomatoid granulomatosis.
本发明还提供本发明所述的化合物或其可药用盐在制备BTK抑制剂中的应用，特别是在制备治疗细胞增生疾病中的应用。所述的细胞增生疾病包括癌症。换言之，本发明还提供通式(I)～通式(V)所述的化合物、及其光学异构体或其药学上可接受的盐或溶剂合物单独或和其他药物联合使用在治疗增生类疾病(如癌症)中的应用。能和本发明所提供的化合物或其可药用盐联合使用的抗肿瘤药包括但并非限定至少一种以下种类：有丝***抑制剂(如长春碱、长春地辛和长春瑞宾)；微管蛋白分解抑制剂(如泰素)；烷基化试剂(如顺铂、卡铂和环磷酰胺)；抗代谢物(如5-氟尿嘧啶、替加氟、甲氨蝶呤、阿糖胞苷和羟基脲)；可***抗生素(如阿雷素、丝裂霉素和争光霉素)；酶(如天门冬氨酶)；拓朴异构酶抑制剂(如依托伯苷和喜树碱)；生物反应调节剂(如干扰素)。The invention also provides the use of a compound of the invention or a pharmaceutically acceptable salt thereof for the preparation of a BTK inhibitor, in particular for the preparation of a medicament for the treatment of a cell proliferative disorder. The cell proliferative diseases include cancer. In other words, the present invention also provides a compound of the formula (I) to the formula (V), and an optical isomer thereof, or a pharmaceutically acceptable salt or solvate thereof, alone or in combination with other drugs for treating hyperplasia Applications in diseases such as cancer. Antineoplastic agents which can be used in combination with a compound provided by the present invention or a pharmaceutically acceptable salt thereof include, but are not limited to, at least one of the following classes: mitotic inhibitors (e.g., vinblastine, vindesine, and vinorelbine); tubulin Decomposition inhibitors (such as Taxol); alkylating agents (such as cisplatin, carboplatin and cyclophosphamide); antimetabolites (such as 5-fluorouracil, tegafur, methotrexate, cytarabine and hydroxyl Urea); insertable antibiotics (such as arrhenone, mitomycin and bleomycin); enzymes (such as aspartate); topoisomerase inhibitors (such as etoricin and camptothecin); biological response regulation Agent (such as interferon).
本发明还提供了制备通式(I’)及其药学可接受衍生物的方法，以如下方案所示的合成路线合成： The present invention also provides a process for the preparation of the general formula (I') and a pharmaceutically acceptable derivative thereof, which is synthesized by the synthetic route shown in the following scheme:
Figure PCTCN2016081669-appb-000012
Figure PCTCN2016081669-appb-000012
如上述合成路线所示，化合物1(参考WO2012158795的方法制备)与R1B(OH)2在磷酸钾、钯催化剂及合适的溶剂或混合溶剂如二氧六环/水存在下，回流反应，得到的化合物2在三苯基磷、DIAD及合适溶剂如THF存在下，与Boc保护的3-羟基哌啶反应得到化合物3，随后在酸性条件下水解制备关键中间体4。该关键中间体DCC及合适溶剂如DCM存在下，与取代丙烯酸片段缩合反应，得到通式(I’)化合物。上式中，R1的结构与上文中通式(I)～通式(V)所述的化合物对应，可根据实际需要选择适当的取代硼酸。例如常用的取代硼酸包括但不限于4-苯氧基苯硼酸、4-(4-甲氧基苯氧基)苯硼酸、4-(3-甲氧基苯氧基)苯硼酸、4-(3,4-二甲氧基苯氧基)苯硼酸、4-(4-氟苯氧基)苯硼酸、4-(4-氯苯氧基)苯硼酸、6-苯氧基吡啶-3-基硼酸、6-(2-氟苯氧基)吡啶-3-基硼酸、6-(4-氟苯氧基)吡啶-3-基硼酸等等。As shown in the above synthetic route, Compound 1 (prepared by the method of WO2012158795) is refluxed with R 1 B(OH) 2 in the presence of potassium phosphate, a palladium catalyst and a suitable solvent or a mixed solvent such as dioxane/water. The obtained compound 2 is reacted with Boc-protected 3-hydroxypiperidine in the presence of triphenylphosphine, DIAD and a suitable solvent such as THF to give compound 3, followed by hydrolysis under acidic conditions to give key intermediate 4. The key intermediate DCC and a suitable solvent such as DCM are condensed with a substituted acrylic acid moiety to give a compound of the formula (I'). In the above formula, the structure of R 1 corresponds to the compound of the above formula (I) to formula (V), and an appropriate substituted boronic acid can be selected according to actual needs. For example, commonly used substituted boronic acids include, but are not limited to, 4-phenoxybenzeneboronic acid, 4-(4-methoxyphenoxy)benzeneboronic acid, 4-(3-methoxyphenoxy)benzeneboronic acid, 4-( 3,4-Dimethoxyphenoxy)benzeneboronic acid, 4-(4-fluorophenoxy)benzeneboronic acid, 4-(4-chlorophenoxy)benzeneboronic acid, 6-phenoxypyridine-3- Boric acid, 6-(2-fluorophenoxy)pyridin-3-ylboronic acid, 6-(4-fluorophenoxy)pyridin-3-ylboronic acid and the like.
本发明发明人通过实验证实，本发明化合物对A549、SGC7901、MCF-7、PC-9、HL-60等肿瘤细胞株具有抗增殖抑制作用，可应用于治疗人或动物细胞增殖性相关的实体瘤或血癌的药物中。The present inventors have confirmed by experiments that the compound of the present invention has an anti-proliferative inhibitory effect on tumor cell lines such as A549, SGC7901, MCF-7, PC-9, HL-60, and the like, and can be applied to the treatment of human or animal cell proliferation-related entities. In cancer or blood cancer drugs.
本发明发明人通过实验证实，本发明化合物具有较好的药代动力学性质，可应用于口服治疗人或动物细胞增殖性相关的实体瘤或血癌或者罹患自身免疫性疾病。The inventors of the present invention have confirmed by experiments that the compound of the present invention has good pharmacokinetic properties and can be applied to orally treating solid tumors or blood cancers associated with cell proliferation of human or animal cells or suffering from autoimmune diseases.
本发明发明人通过实验证实，本发明化合物具有双位点的反应特性。The inventors of the present invention confirmed by experiments that the compound of the present invention has a two-site reaction property.
具体实施方式detailed description
下面通过实施例来说明本发明的可实施性，本领域的技术人员应当理解，根据现有技术的教导，对相应的技术特征进行修改或替换，仍然属于本发明要求保护的范围。The embodiments of the present invention are described below by way of examples, and those skilled in the art should understand that modifications and substitutions of the corresponding technical features are still within the scope of the claimed invention.
实施例1.关键中间体4a的制备Example 1. Preparation of key intermediate 4a
Figure PCTCN2016081669-appb-000013
Figure PCTCN2016081669-appb-000013
步骤1. 3-(4-苯氧基苯基)-1H-吡唑并[3,4-d]嘧啶-4-胺(化合物2a)的合成 Step 1. Synthesis of 3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (Compound 2a)
将3-碘-1H-吡唑并[3,4-d]嘧啶-4-胺(化合物1)(2.61g,10mmol)、4-苯氧基苯硼酸(3.85g,18mmol)和磷酸钾(5.375g,25mmol)依次加入单颈瓶中，加入1,4-二氧六环(40mL)和水(10mL)，氮气保护下，加入三苯基磷钯(1.76g,1.5mmol)，回流反应24h。反应完毕冷却至室温，搅拌过夜，析出黄色沉淀，抽滤，水洗(50ml*3)，干燥24小时后得黄色固体2.18g，为3-(4-苯氧基苯基)-1H-吡唑并[3,4-d]嘧啶-4-胺(化合物2a)，产率72％，LC-ESI-MS：[M+H]＝304.2。3-iodo-1H-pyrazolo[3,4-d]pyrimidin-4-amine (Compound 1) (2.61 g, 10 mmol), 4-phenoxyphenylboronic acid (3.85 g, 18 mmol) and potassium phosphate ( 5.375 g, 25 mmol) was added to a single-necked flask in turn, and 1,4-dioxane (40 mL) and water (10 mL) were added, and under a nitrogen atmosphere, triphenylphosphorous palladium (1.76 g, 1.5 mmol) was added and refluxed. 24h. After completion of the reaction, the mixture was cooled to room temperature, stirred overnight, and a yellow precipitate was precipitated, filtered, washed with water (50ml*3), and dried for 24 hours to give a white solid, 2.18 g, as 3-(4-phenoxyphenyl)-1H-pyrazole. And [3,4-d]pyrimidin-4-amine (Compound 2a), yield 72%, LC-ESI-MS: [M+H]=304.2.
步骤2. 3-(4-氨基-3-(4-苯氧基苯基)-1H-吡唑并[3,4-d]嘧啶-1-基)哌啶-1-甲酸叔丁酯(化合物3a)的合成Step 2. 3-(4-Amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidine-1-carboxylic acid tert-butyl ester ( Synthesis of Compound 3a)
将3-(4-苯氧基苯基)-1H-吡唑并[3,4-d]嘧啶-4-胺(化合物2a)(2.12g,7mmol)、3-羟基哌啶-1-甲酸叔丁酯(1.55g,7.7mmol)、三苯基磷(2.75g,10.5mmol)和偶氮二异丁腈(2.12g,10.5mmol)溶于THF(250ml)中，室温反应12h(TLC薄层层析监测反应是否完全)。反应完毕后，减压回收溶剂。向剩余反应混合物中加入100乙酸乙酯，用100ml*3饱和碳酸钠溶液萃取有机层3次，合并有机相，饱和氯化钠洗1次后，无水硫酸钠干燥。减压回收溶剂得到棕色固体1.12g，产率33％，LC-ESI-MS：[M+H]＝487.3。3-(4-Phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (Compound 2a) (2.12 g, 7 mmol), 3-hydroxypiperidine-1-carboxylic acid tert-Butyl ester (1.55g, 7.7mmol), triphenylphosphine (2.75g, 10.5mmol) and azobisisobutyronitrile (2.12g, 10.5mmol) were dissolved in THF (250ml) and reacted for 12h at room temperature (TLC thin) Layer chromatography monitors whether the reaction is complete). After the reaction was completed, the solvent was recovered under reduced pressure. Ethyl acetate was added to the residue, and the organic layer was extracted three times with 100 ml of aq. The solvent was evaporated under reduced pressure to give a white solid (yield: ield:
步骤3. 3-(4-苯氧基苯基)-1-(哌啶-3-基)-1H-吡唑并[3,4-d]嘧啶-4-胺盐酸盐(化合物4a)的合成Step 3. 3-(4-Phenoxyphenyl)-1-(piperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine hydrochloride (Compound 4a) Synthesis
将3-(4-氨基-3-(4-苯氧基苯基)-1H-吡唑并[3,4-d]嘧啶-1-基)哌啶-1-甲酸叔丁酯(化合物3a)(0.97g,2mmol)溶解于15ml二氧六环，滴加10ml 2N HCl，室温搅拌过夜。减压回收溶剂得到的粗品用甲醇重结晶，得到类白色固体4a(0.56g，产率66％，LC-ESI-MS：[M+H]＝387.1)。3-(4-Amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidine-1-carboxylic acid tert-butyl ester (Compound 3a) (0.97 g, 2 mmol) was dissolved in 15 ml of dioxane, and 10 ml of 2N HCl was added dropwise and stirred at room temperature overnight. The solvent obtained by the solvent was evaporated to give crystals crystals crystals crystals crystals
实施例2.关键中间体4b的制备Example 2. Preparation of key intermediate 4b
Figure PCTCN2016081669-appb-000014
Figure PCTCN2016081669-appb-000014
步骤1. 3-(4-(4-甲氧基苯氧基)苯基)-1H-吡唑并[3,4-d]嘧啶-4-胺(化合物2b)的合成Step 1. Synthesis of 3-(4-(4-methoxyphenoxy)phenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (Compound 2b)
合成步骤参考实施例1步骤1.用类似于化合物2a的合成方法，从4-(4-甲氧基苯氧基)苯硼酸(4.39g,18mmol)制备化合物2b(2.56g，产率77％)，LC-ESI-MS：[M+H]＝334.1。Synthesis procedure Reference Example 1 Step 1. Compound 2b (2.56 g, yield 77%) was prepared from 4-(4-methoxyphenoxy)benzeneboronic acid (4.39 g, 18 mmol) using a procedure similar to compound 2a. ), LC-ESI-MS: [M+H] = 334.1.
步骤2. 3-(4-氨基-3-(4-(4-甲氧基苯氧基)苯基)-1H-吡唑并[3,4-d]嘧啶-1-基)哌啶-1-甲酸叔丁酯(化合物3b)的合成Step 2. 3-(4-Amino-3-(4-(4-methoxyphenoxy)phenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidine- Synthesis of 1-butylic acid tert-butyl ester (compound 3b)
合成步骤参考实施例1步骤2.用类似于化合物3a的合成方法制备化合物3b(1.26g，产率35％)，LC-ESI-MS：[M+H]＝516.2。Synthesis procedure Reference Example 1 Step 2. Compound 3b (1.26 g, yield 35%) was obtained by a procedure similar to compound 3a, LC-ESI-MS: [M+H]=516.2.
步骤3. 3-(4-(4-甲氧基苯氧基)苯基)-1-(哌啶-3-基)-1H-吡唑并[3,4-d]嘧啶-4-胺盐酸盐(化合物4b)的合成Step 3. 3-(4-(4-Methoxyphenoxy)phenyl)-1-(piperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine Synthesis of hydrochloride (Compound 4b)
合成步骤参考实施例1步骤3.用类似于化合物4a的合成方法制备化合物4b(0.59，产率65％，LC-ESI-MS：[M+H]＝417.3)。Synthesis procedure Reference Example 1 Step 3. Compound 4b (0.59, yield 65%, LC-ESI-MS: [M+H] = 417.3).
实施例3.关键中间体4c的制备 Example 3. Preparation of key intermediate 4c
Figure PCTCN2016081669-appb-000015
Figure PCTCN2016081669-appb-000015
步骤1. 3-(4-(3-甲氧基苯氧基)苯基)-1H-吡唑并[3,4-d]嘧啶-4-胺(化合物2c)的合成Step 1. Synthesis of 3-(4-(3-methoxyphenoxy)phenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (Compound 2c)
合成步骤参考实施例1步骤1.用类似于化合物2a的合成方法，从4-(3-甲氧基苯氧基)苯硼酸(4.39g,18mmol)制备化合物2c(2.49g，产率75％，LC-ESI-MS：[M+H]＝334.1)。Synthesis procedure Reference Example 1 Step 1. Compound 2c (2.49 g, yield 75%) was prepared from 4-(3-methoxyphenoxy)benzeneboronic acid (4.39 g, 18 mmol) using a procedure similar to compound 2a. , LC-ESI-MS: [M+H] = 334.1).
步骤2. 3-(4-氨基-3-(4-(3-甲氧基苯氧基)苯基)-1H-吡唑并[3,4-d]嘧啶-1-基)哌啶-1-甲酸叔丁酯(化合物3c)的合成Step 2. 3-(4-Amino-3-(4-(3-methoxyphenoxy)phenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidine- Synthesis of 1-butylic acid tert-butyl ester (compound 3c)
合成步骤参考实施例1步骤2.用类似于化合物3a的合成方法制备化合物3c(1.25g，产率35％)，LC-ESI-MS：[M+H]＝517.2。Synthesis procedure Reference Example 1 Step 2. Compound 3c (1.25 g, yield 35%) was obtained by a procedure similar to compound 3a, LC-ESI-MS: [M+H]=517.2.
步骤3. 3-(4-(3-甲氧基苯氧基)苯基)-1-(哌啶-3-基)-1H-吡唑并[3,4-d]嘧啶-4-胺盐酸盐(化合物4c)的合成Step 3. 3-(4-(3-Methoxyphenoxy)phenyl)-1-(piperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine Synthesis of hydrochloride (Compound 4c)
合成步骤参考实施例1步骤3.用类似于化合物4a的合成方法制备化合物4c(0.61g，产率68％，LC-ESI-MS：[M+H]＝417.3)。Synthesis procedure Reference Example 1 Step 3. Compound 4c (0.61 g, yield 68%, LC-ESI-MS: [M+H] = 417.3) was obtained.
实施例4.关键中间体4d的制备Example 4. Preparation of key intermediate 4d
Figure PCTCN2016081669-appb-000016
Figure PCTCN2016081669-appb-000016
步骤1. 3-(4-(3,4-二甲氧基苯氧基)苯基)-1H-吡唑并[3,4-d]嘧啶-4-胺(化合物2d)的合成Step 1. Synthesis of 3-(4-(3,4-dimethoxyphenoxy)phenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (Compound 2d)
合成步骤参考实施例1步骤1.用类似于化合物2a的合成方法，从4-(3,4-二甲氧基苯氧基)苯硼酸(4.93g,18mmol)制备化合物2d(2.62g，产率72％)，LC-ESI-MS：[M+H]＝367.1。Synthesis procedure Reference Example 1 Step 1. Preparation of compound 2d (2.62 g, from 4-(3,4-dimethoxyphenoxy)benzeneboronic acid (4.93 g, 18 mmol) using a procedure similar to compound 2a. Rate: 72%), LC-ESI-MS: [M+H] = 367.1.
步骤2. 3-(4-氨基-3-(4-(3,4-二甲氧基苯氧基)苯基)-1H-吡唑并[3,4-d]嘧啶-1-基)哌啶-1-甲酸叔丁酯(化合物3d)的合成Step 2. 3-(4-Amino-3-(4-(3,4-dimethoxyphenoxy)phenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl) Synthesis of tert-butyl piperidine-1-carboxylate (compound 3d)
合成步骤参考实施例1步骤2.用类似于化合物3a的合成方法制备化合物3d(1.04g，产率30％)，LC-ESI-MS：[M+H]＝547.3。Synthesis procedure Reference Example 1 Step 2. Compound 3d (1.04 g, yield 30%) was obtained by the procedure of compound 3a, LC-ESI-MS: [M+H]= 547.3.
步骤3. 3-(4-(3,4-二甲氧基苯氧基)苯基)-1-(哌啶-3-基)-1H-吡唑并[3,4-d]嘧啶-4-胺盐酸盐(化合物4d)的合成Step 3. 3-(4-(3,4-Dimethoxyphenoxy)phenyl)-1-(piperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidine- Synthesis of 4-Amine Hydrochloride (Compound 4d)
合成步骤参考实施例1步骤3.用类似于化合物4a的合成方法制备化合物4d(0.53g，产率64％，LC-ESI-MS：[M+H]＝447.1)。 Synthesis procedure Reference Example 1 Step 3. Compound 4d (0.53 g, yield 64%, LC-ESI-MS: [M+H] = 447.1).
实施例5.关键中间体4e的制备Example 5. Preparation of key intermediate 4e
Figure PCTCN2016081669-appb-000017
Figure PCTCN2016081669-appb-000017
步骤1. 3-(4-(4-氟苯氧基)苯基)-1H-吡唑并[3,4-d]嘧啶-4-胺(化合物2e)的合成Step 1. Synthesis of 3-(4-(4-fluorophenoxy)phenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (Compound 2e)
合成步骤参考实施例1步骤1.用类似于化合物2a的合成方法，从4-(4-氟苯氧基)苯硼酸(4.18g,18mmol)制备化合物2e(2.37g，产率74％)，LC-ESI-MS：[M+H]＝322.4。Synthesis procedure Reference Example 1 Step 1. Compound 2e (2.37 g, yield 74%) was prepared from 4-(4-fluorophenoxy)benzeneboronic acid (4.18 g, 18 mmol) using a procedure similar to compound 2a. LC-ESI-MS: [M+H] = 322.4.
步骤2. 3-(4-氨基-3-(4-(4-氟苯氧基)苯基)-1H-吡唑并[3,4-d]嘧啶-1-基)哌啶-1-甲酸叔丁酯(化合物3e)的合成Step 2. 3-(4-Amino-3-(4-(4-fluorophenoxy)phenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1- Synthesis of tert-butyl formate (compound 3e)
合成步骤参考实施例1步骤2.用类似于化合物3a的合成方法制备化合物3e(1.30g,产率37％)，LC-ESI-MS：[M+H]＝504.2。Synthesis procedure Reference Example 1 Step 2. Compound 3e (1.30 g, yield 37%) was obtained using a procedure similar to compound 3a, LC-ESI-MS: [M+H]=504.2.
步骤3. 3-(4-(4-氟苯氧基)苯基)-1-(哌啶-3-基)-1H-吡唑并[3,4-d]嘧啶-4-胺盐酸盐(化合物4e)的合成Step 3. 3-(4-(4-Fluorophenoxy)phenyl)-1-(piperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine hydrochloride Synthesis of salt (compound 4e)
合成步骤参考实施例1步骤3.用类似于化合物4a的合成方法制备化合物4e(0.65g,产率74％，LC-ESI-MS：[M+H]＝405.1)。Synthesis procedure Reference Example 1 Step 3. Compound 4e (0.65 g, yield 74%, LC-ESI-MS: [M+H] = 405.1) was obtained from the compound.
实施例6.关键中间体4f的制备Example 6. Preparation of key intermediate 4f
Figure PCTCN2016081669-appb-000018
Figure PCTCN2016081669-appb-000018
步骤1. 3-(4-(4-氯苯氧基)苯基)-1H-吡唑并[3,4-d]嘧啶-4-胺(化合物2f)的合成Step 1. Synthesis of 3-(4-(4-chlorophenoxy)phenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (Compound 2f)
合成步骤参考实施例1步骤1.用类似于化合物2a的合成方法，从4-(4-氯苯氧基)苯硼酸(4.46g,18mmol)制备化合物2f(2.29g，产率68％)，LC-ESI-MS：[M+H]＝338.1。Synthesis procedure Reference Example 1 Step 1. Compound 2f (2.29 g, yield 68%) was prepared from 4-(4-chlorophenoxy)benzeneboronic acid (4.46 g, 18 mmol) using a procedure similar to compound 2a. LC-ESI-MS: [M+H] =353.
步骤2. 3-(4-氨基-3-(4-(4-氯苯氧基)苯基)-1H-吡唑并[3,4-d]嘧啶-1-基)哌啶-1-甲酸叔丁酯(化合物3f)的合成Step 2. 3-(4-Amino-3-(4-(4-chlorophenoxy)phenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1- Synthesis of tert-butyl formate (compound 3f)
合成步骤参考实施例1步骤2.用类似于化合物3a的合成方法制备化合物3f(1.46g,产率40％)，LC-ESI-MS：[M+H]＝521.2。Synthesis procedure Reference Example 1 Step 2. Compound 3f (1.46 g, yield 40%) was obtained by the procedure of compound 3a, LC-ESI-MS: [M+H]=521.2.
步骤3. 3-(4-(4-氯苯氧基)苯基)-1-(哌啶-3-基)-1H-吡唑并[3,4-d]嘧啶-4-胺盐酸盐(化合物4f)的合成Step 3. 3-(4-(4-Chlorophenoxy)phenyl)-1-(piperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine hydrochloride Synthesis of salt (compound 4f)
合成步骤参考实施例1步骤3.用类似于化合物4a的合成方法制备化合物4f(0.72g,产率79％，[M+H]＝421.2)。Synthesis procedure Reference Example 1 Step 3. Compound 4f (0.72 g, yield 79%, [M+H] = 421.2) was obtained by a procedure similar to compound 4a.
实施例7. 1-(3-(4-氨基-3-(4-苯氧基苯基)-1H-吡唑并[3,4-d]嘧啶-1-基)哌啶-1-基)-2-氟丙基 -2-烯-1-酮的制备(化合物5-1)Example 7. 1-(3-(4-Amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl -2-fluoropropyl Preparation of 2-en-1-one (Compound 5-1)
Figure PCTCN2016081669-appb-000019
Figure PCTCN2016081669-appb-000019
化合物4a(548mg,1.3mmol)，2-氟丙烯酸(117mg,1.3mmol)溶于5ml二氯甲烷，室温滴加DCC(267mg,1.3mmol)的二氯甲烷(5ml)溶液，回流反应4小时。反应结束后，冷却至室温，抽滤，滤液浓缩后，柱层析分离(石油醚：乙酸乙酯：三乙胺＝2：1：0.1)得白色固体207mg，产率35％，LC-ESI-MS：[M+H]＝459.2。Compound 4a (548 mg, 1.3 mmol), 2-fluoroacrylic acid (117 mg, 1.3 mmol) was dissolved in dichloromethane (5 ml), and a solution of DCC (267 mg, 1.3 mmol) in dichloromethane (5 ml). After completion of the reaction, the mixture was cooled to room temperature, filtered, filtered, and then filtered, and then evaporated to ethylamine (ethyl ether: ethyl acetate: triethylamine = 2:1:0.1) - MS: [M+H] = 459.2.
1H-NMR(δ,CDCl3):8.34(s,1H,ArH),7.65(d,2H,J＝8.0Hz,ArH),7.39(m,2H,ArH),7.16(m,3H,ArH),7.08(d,2H,J＝8.0Hz,ArH),6.12(s,2H,-NH2),5.30(d,0.5H,J＝4.0Hz,-C＝CH2),5.18(d,0.5H,J＝4.0Hz,-C＝CH2),5.10(d,1H,-C＝CH2),4.92(m,1H,piperidine),4.66(m,0.5H,piperidine),4.44(m,0.5H,piperidine),4.12(m,0.5H,piperidine),3.76(m,0.5H,piperidine),3.47(m,0.5H,piperidine),3.17(m,0.5H,piperidine),2.87(m,0.5H,piperidine),2.81(m,0.5H,piperidine),2.30(m,2H,piperidine),1.75(m,2H,piperidine). 1 H-NMR (δ, CDCl 3 ): 8.34 (s, 1H, ArH), 7.65 (d, 2H, J = 8.0 Hz, ArH), 7.39 (m, 2H, ArH), 7.16 (m, 3H, ArH) ), 7.08 (d, 2H, J = 8.0 Hz, ArH), 6.12 (s, 2H, -NH 2 ), 5.30 (d, 0.5H, J = 4.0 Hz, -C = CH 2 ), 5.18 (d, 0.5H, J=4.0 Hz, -C=CH 2 ), 5.10 (d, 1H, -C=CH 2 ), 4.92 (m, 1H, piperididine), 4.66 (m, 0.5H, piperididine), 4.44 (m) , 0.5H, piperidine), 4.12 (m, 0.5H, piperididine), 3.76 (m, 0.5H, piperididine), 3.47 (m, 0.5H, piperididine), 3.17 (m, 0.5H, piperididine), 2.87 (m) , 0.5H, piperidine), 2.81 (m, 0.5H, piperididine), 2.30 (m, 2H, piperididine), 1.75 (m, 2H, piperididine).
实施例8. 1-(3-(4-氨基-3-(4-苯氧基苯基)-1H-吡唑并[3,4-d]嘧啶-1-基)哌啶-1-基)-2-氯丙基-2-烯-1-酮的制备(化合物5-2)Example 8. 1-(3-(4-Amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl Preparation of 2-chloropropyl-2-en-1-one (Compound 5-2)
Figure PCTCN2016081669-appb-000020
Figure PCTCN2016081669-appb-000020
合成步骤参考实施例7.用类似于化合物5-1的合成方法，将2-氟丙烯酸替换为2-氯丙烯酸制备化合物5-2(257mg，产率42％，LC-ESI-MS：[M+H]＝475.2)。Synthesis procedure Reference Example 7. Compound 5-2 (257 mg, yield 42%, LC-ESI-MS: [M. +H]=475.2).
1H-NMR(δ,CDCl3):8.37(s,1H,ArH),7.65(d,2H,J＝8.0Hz,ArH),7.40(m,2H,ArH),7.17(m,3H,ArH),7.09(d,2H,J＝8.0Hz,ArH),5.67(s,2H,-NH2),5.62(m,2H,-C＝CH2),4.93(m,1H,piperidine),4.68(m,0.5H,piperidine),4.52(m,0.5H,piperidine),4.13(m,0.5H,piperidine),3.98(m,0.5H,piperidine),3.80(m,0.5H,piperidine),3.50(m,0.5H,piperidine),3.23(m,0.5H,piperidine),2.91(m,0.5H,piperidine),2.30(m,2H,piperidine),1.77(m,2H,piperidine). 1 H-NMR (δ, CDCl 3 ): 8.37 (s, 1H, ArH), 7.65 (d, 2H, J = 8.0 Hz, ArH), 7.40 (m, 2H, ArH), 7.17 (m, 3H, ArH) ), 7.09 (d, 2H, J = 8.0 Hz, ArH), 5.67 (s, 2H, -NH 2 ), 5.62 (m, 2H, -C = CH 2 ), 4.93 (m, 1H, piperididine), 4.68 (m, 0.5H, piperidine), 4.52 (m, 0.5H, piperididine), 4.13 (m, 0.5H, piperididine), 3.98 (m, 0.5H, piperididine), 3.80 (m, 0.5H, piperididine), 3.50 (m, 0.5H, piperididine), 3.23 (m, 0.5H, piperididine), 2.91 (m, 0.5H, piperididine), 2.30 (m, 2H, piperididine), 1.77 (m, 2H, piperididine).
实施例9. 1-(3-(4-氨基-3-(4-苯氧基苯基)-1H-吡唑并[3,4-d]嘧啶-1-基)哌啶-1-基)-2-溴丙基-2-烯-1-酮的制备(化合物5-3) Example 9. 1-(3-(4-Amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl Preparation of 2-bromopropyl-2-en-1-one (Compound 5-3)
Figure PCTCN2016081669-appb-000021
Figure PCTCN2016081669-appb-000021
合成步骤参考实施例7.用类似于化合物5-1的合成方法，将2-氟丙烯酸替换为2-溴丙烯酸制备化合物5-3(225mg，产率33％，LC-ESI-MS：[M+H]＝519.2)。Synthesis procedure Reference Example 7. Compound 5-3 (225 mg, yield 33%, LC-ESI-MS: [M. +H]=519.2).
1H-NMR(δ,CDCl3):8.37(s,1H,ArH),7.64(d,2H,J＝8.0Hz,ArH),7.40(m,2H,ArH),7.16(m,3H,ArH),7.09(d,2H,J＝8.0Hz,ArH),6.05(s,1H,-NH2),5.79(m,3H,-C＝CH2,-NH2),4.93(m,1H,piperidine),4.66(m,0.5H,piperidine),4.51(m,0.5H,piperidine),4.11(m,0.5H,piperidine),3.97(m,0.5H,piperidine),3.77(m,0.5H,piperidine),3.22(m,0.5H,piperidine),3.11(m,0.5H,piperidine),2.91(m,0.5H,piperidine),2.28(m,2H,piperidine),1.71(m,2H,piperidine). 1 H-NMR (δ, CDCl 3 ): 8.37 (s, 1H, ArH), 7.64 (d, 2H, J = 8.0 Hz, ArH), 7.40 (m, 2H, ArH), 7.16 (m, 3H, ArH) ), 7.09 (d, 2H, J = 8.0 Hz, ArH), 6.05 (s, 1H, -NH 2 ), 5.79 (m, 3H, -C=CH 2 , -NH 2 ), 4.93 (m, 1H, Piperidine), 4.66 (m, 0.5H, piperidine), 4.51 (m, 0.5H, piperididine), 4.11 (m, 0.5H, piperididine), 3.97 (m, 0.5H, piperididine), 3.77 (m, 0.5H, Piperidine), 3.22 (m, 0.5H, piperidine), 3.11 (m, 0.5H, piperididine), 2.91 (m, 0.5H, piperididine), 2.28 (m, 2H, piperididine), 1.71 (m, 2H, piperididine) .
实施例10. 1-(3-(4-氨基-3-(4-(4-甲氧基苯氧基)苯基)-1H-吡唑并[3,4-d]嘧啶-1-基)哌啶-1-基)-2-氟丙基-2-烯-1-酮的制备(化合物5-4)Example 10. 1-(3-(4-Amino-3-(4-(4-methoxyphenoxy)phenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl Preparation of piperidin-1-yl)-2-fluoropropyl-2-en-1-one (Compound 5-4)
Figure PCTCN2016081669-appb-000022
Figure PCTCN2016081669-appb-000022
合成步骤参考实施例7.用类似于化合物5-1的合成方法，将化合物4a替换为化合物4b，制备化合物5-4(212mg，产率33％，LC-ESI-MS：[M+H]＝489.2)。Synthesis procedure Reference Example 7. Compound 5a was prepared by substituting compound 4a with compound 4b to give compound 5b (212 mg, yield 33%, LC-ESI-MS: [M+H] =489.2).
1H-NMR(δ,CDCl3):8.35(s,1H,ArH),7.66(d,2H,J＝8.0Hz,ArH),7.38(d,2H,J＝8.0Hz,ArH),7.17(d,2H,J＝8.0Hz,ArH),7.08(d,2H,J＝8.0Hz,ArH),6.15(s,2H,-NH2),5.31(d,0.5H,J＝4.0Hz,-C＝CH2),5.18(d,0.5H,J＝4.0Hz,-C＝CH2),5.10(d,1H,-C＝CH2),4.92(m,1H,piperidine),4.66(m,0.5H,piperidine),4.44(m,0.5H,piperidine),4.12(m,0.5H,piperidine),3.76(m,0.5H,piperidine),3.62(s,3H,-OCH3),3.47(m,0.5H,piperidine),3.17(m,0.5H,piperidine),2.87(m,0.5H,piperidine),2.81(m,0.5H,piperidine),2.30(m,2H,piperidine),1.75(m,2H,piperidine). 1 H-NMR (δ, CDCl 3 ): 8.35 (s, 1H, ArH), 7.66 (d, 2H, J = 8.0 Hz, ArH), 7.38 (d, 2H, J = 8.0 Hz, ArH), 7.17 ( d, 2H, J = 8.0 Hz, ArH), 7.08 (d, 2H, J = 8.0 Hz, ArH), 6.15 (s, 2H, -NH 2 ), 5.31 (d, 0.5H, J = 4.0 Hz, - C=CH 2 ), 5.18 (d, 0.5H, J=4.0 Hz, -C=CH 2 ), 5.10 (d, 1H, -C=CH 2 ), 4.92 (m, 1H, piperididine), 4.66 (m) , 0.5H, piperidine), 4.44 (m, 0.5H, piperididine), 4.12 (m, 0.5H, piperididine), 3.76 (m, 0.5H, piperididine), 3.62 (s, 3H, -OCH 3 ), 3.47 ( m, 0.5H, piperidine), 3.17 (m, 0.5H, piperididine), 2.87 (m, 0.5H, piperididine), 2.81 (m, 0.5H, piperididine), 2.30 (m, 2H, piperididine), 1.75 (m) , 2H, piperidine).
实施例11. 1-(3-(4-氨基-3-(4-(4-甲氧基苯氧基)苯基)-1H-吡唑并[3,4-d]嘧啶-1-基)哌啶-1-基)-2-氯丙基-2-烯-1-酮的制备(化合物5-5) Example 11. 1-(3-(4-Amino-3-(4-(4-methoxyphenoxy)phenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl Preparation of piperidin-1-yl)-2-chloropropyl-2-en-1-one (Compound 5-5)
Figure PCTCN2016081669-appb-000023
Figure PCTCN2016081669-appb-000023
合成步骤参考实施例7.用类似于化合物5-1的合成方法，将化合物4a替换为化合物4b，将2-氟丙烯酸替换为2-氯丙烯酸，制备化合物5-5(212mg，产率38％，LC-ESI-MS：[M+H]＝505.1)。Synthesis procedure Reference Example 7. Compound 5a (212 mg, yield 38%) was prepared by a procedure similar to compound 5-1, substituting compound 4a for compound 4b and 2-fluoroacrylic acid for 2-chloroacrylic acid. , LC-ESI-MS: [M+H] = 505.1).
1H-NMR(δ,CDCl3):8.37(s,1H,ArH),7.65(d,2H,J＝8.0Hz,ArH),7.38(d,2H,J＝8.0Hz,ArH),7.17(d,2H,J＝8.0Hz,ArH),7.09(d,2H,J＝8.0Hz,ArH),6.03(s,2H,-NH2),5.24(d,0.5H,J＝4.0Hz,-C＝CH2),5.07(d,0.5H,J＝4.0Hz,-C＝CH2),5.01(d,1H,-C＝CH2),4.87(m,1H,piperidine),4.62(m,0.5H,piperidine),4.39(m,0.5H,piperidine),4.22(m,0.5H,piperidine),3.77(m,0.5H,piperidine),3.68(s,3H,-OCH3),3.45(m,0.5H,piperidine),3.14(m,0.5H,piperidine),2.86(m,0.5H,piperidine),2.78(m,0.5H,piperidine),2.28(m,2H,piperidine),1.74(m,2H,piperidine). 1 H-NMR (δ, CDCl 3 ): 8.37 (s, 1H, ArH), 7.65 (d, 2H, J = 8.0 Hz, ArH), 7.38 (d, 2H, J = 8.0 Hz, ArH), 7.17 ( d, 2H, J = 8.0 Hz, ArH), 7.09 (d, 2H, J = 8.0 Hz, ArH), 6.03 (s, 2H, -NH 2 ), 5.24 (d, 0.5H, J = 4.0 Hz, - C=CH 2 ), 5.07 (d, 0.5H, J=4.0 Hz, -C=CH 2 ), 5.01 (d, 1H, -C=CH 2 ), 4.87 (m, 1H, piperididine), 4.62 (m) , 0.5H, piperidine), 4.39 (m, 0.5H, piperididine), 4.22 (m, 0.5H, piperididine), 3.77 (m, 0.5H, piperididine), 3.68 (s, 3H, -OCH 3 ), 3.45 ( m, 0.5H, piperidine), 3.14 (m, 0.5H, piperididine), 2.86 (m, 0.5H, piperididine), 2.78 (m, 0.5H, piperididine), 2.28 (m, 2H, piperididine), 1.74 (m) , 2H, piperidine).
实施例12. 1-(3-(4-氨基-3-(4-(4-甲氧基苯氧基)苯基)-1H-吡唑并[3,4-d]嘧啶-1-基)哌啶-1-基)-2-溴丙基-2-烯-1-酮的制备(化合物5-6)Example 12. 1-(3-(4-Amino-3-(4-(4-methoxyphenoxy)phenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl Preparation of piperidin-1-yl)-2-bromopropyl-2-en-1-one (compound 5-6)
Figure PCTCN2016081669-appb-000024
Figure PCTCN2016081669-appb-000024
合成步骤参考实施例7.用类似于化合物5-1的合成方法，将化合物4a替换为化合物4b，将2-氟丙烯酸替换为2-溴丙烯酸，制备化合物5-6(234mg，产率33％，LC-ESI-MS：[M+H]＝551.1)。Synthesis procedure Reference Example 7. Compound 5a (234 mg, yield 33%) was prepared by a procedure similar to compound 5-1, substituting compound 4a for compound 4b and 2-fluoroacrylic acid for 2-bromoacrylic acid. , LC-ESI-MS: [M+H] = 551.1).
1H-NMR(δ,CDCl3):8.36(s,1H,ArH),7.66(d,2H,J＝8.0Hz,ArH),7.39(d,2H,J＝8.0Hz,ArH),7.17(d,2H,J＝8.0Hz,ArH),7.09(d,2H,J＝8.0Hz,ArH),6.18(s,2H,-NH2),5.33(d,0.5H,J＝4.0Hz,-C＝CH2),5.19(d,0.5H,J＝4.0Hz,-C＝CH2),5.12(d,1H,-C＝CH2),4.96(m,1H,piperidine),4.68(m,0.5H,piperidine),4.45(m,0.5H,piperidine),4.16(m,0.5H,piperidine),3.77(m,0.5H,piperidine),3.65(s,3H,-OCH3),3.48(m,0.5H,piperidine),3.18(m,0.5H,piperidine),2.87(m,0.5H,piperidine),2.81(m,0.5H,piperidine),2.31(m,2H,piperidine),1.75(m,2H,piperidine). 1 H-NMR (δ, CDCl 3 ): 8.36 (s, 1H, ArH), 7.66 (d, 2H, J = 8.0 Hz, ArH), 7.39 (d, 2H, J = 8.0 Hz, ArH), 7.17 ( d, 2H, J = 8.0 Hz, ArH), 7.09 (d, 2H, J = 8.0 Hz, ArH), 6.18 (s, 2H, -NH 2 ), 5.33 (d, 0.5H, J = 4.0 Hz, - C=CH 2 ), 5.19 (d, 0.5H, J=4.0 Hz, -C=CH 2 ), 5.12 (d, 1H, -C=CH 2 ), 4.96 (m, 1H, piperididine), 4.68 (m) , 0.5H, piperidine), 4.45 (m, 0.5H, piperididine), 4.16 (m, 0.5H, piperididine), 3.77 (m, 0.5H, piperididine), 3.65 (s, 3H, -OCH 3 ), 3.48 ( m, 0.5H, piperidine), 3.18 (m, 0.5H, piperididine), 2.87 (m, 0.5H, piperididine), 2.81 (m, 0.5H, piperididine), 2.31 (m, 2H, piperididine), 1.75 (m) , 2H, piperidine).
实施例13. 1-(3-(4-氨基-3-(4-(3-甲氧基苯氧基)苯基)-1H-吡唑并[3,4-d]嘧啶-1-基)哌啶-1- 基)-2-氟丙基-2-烯-1-酮的制备(化合物5-7)Example 13. 1-(3-(4-Amino-3-(4-(3-methoxyphenoxy)phenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl Piperidine-1- Preparation of 2-(2-fluoropropyl-2-en-1-one) (Compound 5-7)
Figure PCTCN2016081669-appb-000025
Figure PCTCN2016081669-appb-000025
合成步骤参考实施例7.用类似于化合物5-1的合成方法，将化合物4a替换为4c，制备化合物5-7(205mg，产率32％，LC-ESI-MS：[M+H]＝489.2)。Synthesis procedure Reference Example 7. Compound 5a was prepared by substituting compound 4a to 4c using a synthetic procedure analogous to compound 5-1 (205 mg, yield 32%, LC-ESI-MS: [M+H] = 489.2).
1H-NMR(δ,CDCl3):8.37(s,1H,ArH),7.63(d,2H,J＝8.0Hz,ArH),7.12(m,1H,ArH),7.05(d,2H,J＝8.0Hz,ArH),6.52(m,2H,ArH),6.39(s,1H,ArH),6.14(s,2H,-NH2),5.27(d,0.5H,J＝4.0Hz,-C＝CH2),5.06(d,0.5H,J＝4.0Hz,-C＝CH2),5.03(d,1H,-C＝CH2),4.91(m,1H,piperidine),4.62(m,0.5H,piperidine),4.40(m,0.5H,piperidine),4.11(m,0.5H,piperidine),3.73(m,0.5H,piperidine),3.64(s,3H,-OCH3),3.43(m,0.5H,piperidine),3.13(m,0.5H,piperidine),2.87(m,0.5H,piperidine),2.80(m,0.5H,piperidine),2.17(m,2H,piperidine),1.77(m,2H,piperidine). 1 H-NMR (δ, CDCl 3 ): 8.37 (s, 1H, ArH), 7.63 (d, 2H, J = 8.0 Hz, ArH), 7.12 (m, 1H, ArH), 7.05 (d, 2H, J) = 8.0 Hz, ArH), 6.52 (m, 2H, ArH), 6.39 (s, 1H, ArH), 6.14 (s, 2H, -NH 2 ), 5.27 (d, 0.5H, J = 4.0 Hz, -C =CH 2 ), 5.06 (d, 0.5H, J = 4.0 Hz, -C=CH 2 ), 5.03 (d, 1H, -C=CH 2 ), 4.91 (m, 1H, piperididine), 4.62 (m, 0.5H,piperidine), 4.40 (m, 0.5H, piperididine), 4.11 (m, 0.5H, piperididine), 3.73 (m, 0.5H, piperididine), 3.64 (s, 3H, -OCH 3 ), 3.43 (m) , 0.5H, piperidine), 3.13 (m, 0.5H, piperididine), 2.87 (m, 0.5H, piperididine), 2.80 (m, 0.5H, piperididine), 2.17 (m, 2H, piperididine), 1.77 (m, 2H, piperidine).
实施例14. 1-(3-(4-氨基-3-(4-(3-甲氧基苯氧基)苯基)-1H-吡唑并[3,4-d]嘧啶-1-基)哌啶-1-基)-2-氯丙基-2-烯-1-酮的制备(化合物5-8)Example 14. 1-(3-(4-Amino-3-(4-(3-methoxyphenoxy)phenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl Preparation of piperidin-1-yl)-2-chloropropyl-2-en-1-one (compound 5-8)
Figure PCTCN2016081669-appb-000026
Figure PCTCN2016081669-appb-000026
合成步骤参考实施例7.用类似于化合物5-1的合成方法，将化合物4a替换为4c，将2-氟丙烯酸替换为2-氯丙烯酸，制备化合物5-8(272mg，产率41％，LC-ESI-MS：[M+H]＝505.2)。Synthesis procedure Referring to Example 7. Compound 5a (272 mg, yield 41%, was prepared by a procedure similar to compound 5-1, substituting compound 4a to 4c and 2-fluoroacrylic acid to 2-chloroacrylic acid. LC-ESI-MS: [M+H] = 505.2).
1H-NMR(δ,CDCl3):8.36(s,1H,ArH),7.64(d,2H,J＝8.0Hz,ArH),7.16(m,1H,ArH),7.05(d,2H,J＝8.0Hz,ArH),6.53(m,2H,ArH),6.38(s,1H,ArH),6.14(s,2H,-NH2),5.32(d,0.5H,J＝4.0Hz,-C＝CH2),5.16(d,0.5H,J＝4.0Hz,-C＝CH2),5.10(d,1H,-C＝CH2),4.92(m,1H,piperidine),4.63(m,0.5H,piperidine),4.40(m,0.5H,piperidine),4.12(m,0.5H,piperidine),3.73(m,0.5H,piperidine),3.66(s,3H,-OCH3),3.43(m,0.5H,piperidine),3.13(m,0.5H,piperidine),2.85(m,0.5H,piperidine),2.83(m,0.5H,piperidine),2.27(m,2H,piperidine),1.76(m,2H,piperidine). 1 H-NMR (δ, CDCl 3 ): 8.36 (s, 1H, ArH), 7.64 (d, 2H, J = 8.0 Hz, ArH), 7.16 (m, 1H, ArH), 7.05 (d, 2H, J) = 8.0 Hz, ArH), 6.53 (m, 2H, ArH), 6.38 (s, 1H, ArH), 6.14 (s, 2H, -NH 2 ), 5.32 (d, 0.5H, J = 4.0 Hz, -C =CH 2 ), 5.16 (d, 0.5H, J = 4.0 Hz, -C=CH 2 ), 5.10 (d, 1H, -C=CH 2 ), 4.92 (m, 1H, piperididine), 4.63 (m, 0.5H,piperidine), 4.40 (m, 0.5H, piperidine), 4.12 (m, 0.5H, piperididine), 3.73 (m, 0.5H, piperididine), 3.66 (s, 3H, -OCH 3 ), 3.43 (m) , 0.5H, piperidine), 3.13 (m, 0.5H, piperididine), 2.85 (m, 0.5H, piperididine), 2.83 (m, 0.5H, piperididine), 2.27 (m, 2H, piperididine), 1.76 (m, 2H, piperidine).
实施例15. 1-(3-(4-氨基-3-(4-(3-甲氧基苯氧基)苯基)-1H-吡唑并[3,4-d]嘧啶-1-基)哌啶-1-基)-2-溴丙基-2-烯-1-酮的制备(化合物5-9) Example 15. 1-(3-(4-Amino-3-(4-(3-methoxyphenoxy)phenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl Preparation of piperidin-1-yl)-2-bromopropyl-2-en-1-one (compounds 5-9)
Figure PCTCN2016081669-appb-000027
Figure PCTCN2016081669-appb-000027
合成步骤参考实施例7.用类似于化合物5-1的合成方法，将化合物4a替换为4c，将2-氟丙烯酸替换为2-溴丙烯酸，制备化合物5-9(222mg，产率31％，LC-ESI-MS：[M+H]＝551.1)。Synthesis procedure Reference Example 7. Compound 5a (222 mg, yield 31%, was prepared by a procedure similar to compound 5-1, substituting compound 4a to 4c and 2-fluoroacrylic acid to 2-bromoacrylic acid. LC-ESI-MS: [M+H] = 551.1.
1H-NMR(δ,CDCl3):8.36(s,1H,ArH),7.64(d,2H,J＝8.0Hz,ArH),7.13(m,1H,ArH),7.05(d,2H,J＝8.0Hz,ArH),6.52(m,2H,ArH),6.38(s,1H,ArH),6.14(s,2H,-NH2),5.27(d,0.5H,J＝4.0Hz,-C＝CH2),5.10(d,0.5H,J＝4.0Hz,-C＝CH2),5.03(d,1H,-C＝CH2),4.91(m,1H,piperidine),4.62(m,0.5H,piperidine),4.40(m,0.5H,piperidine),4.11(m,0.5H,piperidine),3.76(m,0.5H,piperidine),3.67(s,3H,-OCH3),3.44(m,0.5H,piperidine),3.13(m,0.5H,piperidine),2.85(m,0.5H,piperidine),2.77(m,0.5H,piperidine),2.33(m,2H,piperidine),1.77(m,2H,piperidine). 1 H-NMR (δ, CDCl 3 ): 8.36 (s, 1H, ArH), 7.64 (d, 2H, J = 8.0 Hz, ArH), 7.13 (m, 1H, ArH), 7.05 (d, 2H, J) = 8.0 Hz, ArH), 6.52 (m, 2H, ArH), 6.38 (s, 1H, ArH), 6.14 (s, 2H, -NH 2 ), 5.27 (d, 0.5H, J = 4.0 Hz, -C =CH 2 ), 5.10 (d, 0.5H, J = 4.0 Hz, -C=CH 2 ), 5.03 (d, 1H, -C=CH 2 ), 4.91 (m, 1H, piperididine), 4.62 (m, 0.5H,piperidine), 4.40 (m, 0.5H, piperididine), 4.11 (m, 0.5H, piperididine), 3.76 (m, 0.5H, piperididine), 3.67 (s, 3H, -OCH 3 ), 3.44 (m) , 0.5H, piperidine), 3.13 (m, 0.5H, piperididine), 2.85 (m, 0.5H, piperididine), 2.77 (m, 0.5H, piperididine), 2.33 (m, 2H, piperididine), 1.77 (m, 2H, piperidine).
实施例16. 1-(3-(4-氨基-3-(4-(3,4-二甲氧基苯氧基)苯基)-1H-吡唑并[3,4-d]嘧啶-1-基)哌啶-1-基)-2-氟丙基-2-烯-1-酮的制备(化合物5-10)Example 16. 1-(3-(4-Amino-3-(4-(3,4-dimethoxyphenoxy)phenyl)-1H-pyrazolo[3,4-d]pyrimidine- Preparation of 1-yl)piperidin-1-yl)-2-fluoropropyl-2-en-1-one (Compound 5-10)
Figure PCTCN2016081669-appb-000028
Figure PCTCN2016081669-appb-000028
合成步骤参考实施例7.用类似于化合物5-1的合成方法，将化合物4a替换为4d，制备化合物5-10(211mg，产率31％，LC-ESI-MS：[M+H]＝519.2)。Synthesis procedure Reference Example 7. Compound 5a was prepared by substituting compound 4a to 4d using a synthetic procedure analogous to compound 5-1 (211 mg, yield 31%, LC-ESI-MS: [M+H] = 519.2).
1H-NMR(δ,CDCl3:8.37(s,1H,ArH),7.63(d,2H,J＝8.0Hz,ArH),7.05(d,2H,J＝8.0Hz,ArH),6.62(d,1H,J＝8.0Hz,ArH),6.41(d,1H,J＝8.0Hz,ArH),6.27(s,1H,ArH),5.96(s,2H,-NH2),5.27(d,0.5H,J＝4.0Hz,-C＝CH2),5.06(d,0.5H,J＝4.0Hz,-C＝CH2),5.03(d,1H,-C＝CH2),4.91(m,1H,piperidine),4.62(m,0.5H,piperidine),4.40(m,0.5H,piperidine),4.11(m,0.5H,piperidine),3.74(m,3.5H,-OCH3,piperidine),3.66(s,3H,-OCH3),3.43(m,0.5H,piperidine),3.12(m,0.5H,piperidine),2.87(m,0.5H,piperidine),2.85(m,0.5H,piperidine),2.17(m,2H,piperidine),1.75(m,2H,piperidine). 1 H-NMR (δ, CDCl 3 : 8.37 (s, 1H, ArH), 7.63 (d, 2H, J = 8.0 Hz, ArH), 7.05 (d, 2H, J = 8.0 Hz, ArH), 6.62 (d , 1H, J = 8.0 Hz, ArH), 6.41 (d, 1H, J = 8.0 Hz, ArH), 6.27 (s, 1H, ArH), 5.96 (s, 2H, -NH 2 ), 5.27 (d, 0.5) H, J = 4.0 Hz, -C = CH 2 ), 5.06 (d, 0.5H, J = 4.0 Hz, -C = CH 2 ), 5.03 (d, 1H, -C = CH 2 ), 4.91 (m, 1H,piperidine), 4.62 (m, 0.5H, piperididine), 4.40 (m, 0.5H, piperididine), 4.11 (m, 0.5H, piperididine), 3.74 (m, 3.5H, -OCH 3 , piperididine), 3.66 (s, 3H, -OCH 3 ), 3.43 (m, 0.5H, piperididine), 3.12 (m, 0.5H, piperididine), 2.87 (m, 0.5H, piperididine), 2.85 (m, 0.5H, piperididine), 2.17 (m, 2H, piperididine), 1.75 (m, 2H, piperididine).
实施例17. 1-(3-(4-氨基-3-(4-(3,4-二甲氧基苯氧基)苯基)-1H-吡唑并[3,4-d]嘧啶-1-基)哌啶 -1-基)-2-氯丙基-2-烯-1-酮的制备(化合物5-11)Example 17. 1-(3-(4-Amino-3-(4-(3,4-dimethoxyphenoxy)phenyl)-1H-pyrazolo[3,4-d]pyrimidine- 1-yl) piperidine Preparation of 1-yl)-2-chloropropyl-2-en-1-one (Compound 5-11)
Figure PCTCN2016081669-appb-000029
Figure PCTCN2016081669-appb-000029
合成步骤参考实施例7.用类似于化合物5-1的合成方法，将化合物4a替换为4d，将2-氟丙烯酸替换为2-氯丙烯酸，制备化合物5-11(232mg，产率33％，LC-ESI-MS：[M+H]＝535.2)。Synthesis procedure Reference Example 7. Compound 5a (232 mg, yield 33%, was prepared by a procedure similar to compound 5-1, substituting compound 4a for 4d and 2-fluoroacrylic acid for 2-chloroacrylic acid. LC-ESI-MS: [M+H] = 535.2.
1H-NMR(δ,CDCl3-):8.37(s,1H,ArH),7.64(d,2H,J＝8.0Hz,ArH),7.04(d,2H,J＝8.0Hz,ArH),6.62(d,1H,J＝8.0Hz,ArH),6.42(d,1H,J＝8.0Hz,ArH),6.27(s,1H,ArH),5.96(s,2H,-NH2),5.27(d,0.5H,J＝4.0Hz,-C＝CH2),5.06(d,0.5H,J＝4.0Hz,-C＝CH2),5.06(d,1H,-C＝CH2),4.92(m,1H,piperidine),4.61(m,0.5H,piperidine),4.40(m,0.5H,piperidine),4.11(m,0.5H,piperidine),3.75(m,3.5H,-OCH3,piperidine),3.66(s,3H,-OCH3),3.43(m,0.5H,piperidine),3.12(m,0.5H,piperidine),2.87(m,0.5H,piperidine),2.75(m,0.5H,piperidine),2.31(m,2H,piperidine),1.73(m,2H,piperidine). 1 H-NMR (δ, CDCl 3 -): 8.37 (s, 1H, ArH), 7.64 (d, 2H, J = 8.0 Hz, ArH), 7.04 (d, 2H, J = 8.0 Hz, ArH), 6.62 (d, 1H, J = 8.0 Hz, ArH), 6.42 (d, 1H, J = 8.0 Hz, ArH), 6.27 (s, 1H, ArH), 5.96 (s, 2H, -NH 2 ), 5.27 (d , 0.5H, J=4.0Hz, -C=CH 2 ), 5.06 (d, 0.5H, J=4.0Hz, -C=CH 2 ), 5.06(d,1H,-C=CH 2 ), 4.92( m,1H,piperidine), 4.61 (m,0.5H,piperidine), 4.40 (m,0.5H,piperidine),4.11 (m,0.5H,piperidine),3.75 (m,3.5H,-OCH 3 ,piperidine) , 3.66 (s, 3H, -OCH 3 ), 3.43 (m, 0.5H, piperididine), 3.12 (m, 0.5H, piperididine), 2.87 (m, 0.5H, piperididine), 2.75 (m, 0.5H, piperine ), 2.31 (m, 2H, piperididine), 1.73 (m, 2H, piperididine).
实施例18. 1-(3-(4-氨基-3-(4-(3,4-二甲氧基苯氧基)苯基)-1H-吡唑并[3,4-d]嘧啶-1-基)哌啶-1-基)-2-溴丙基-2-烯-1-酮的制备(化合物5-12)Example 18. 1-(3-(4-Amino-3-(4-(3,4-dimethoxyphenoxy)phenyl)-1H-pyrazolo[3,4-d]pyrimidine- Preparation of 1-yl)piperidin-1-yl)-2-bromopropyl-2-en-1-one (Compound 5-12)
Figure PCTCN2016081669-appb-000030
Figure PCTCN2016081669-appb-000030
合成步骤参考实施例7.用类似于化合物5-1的合成方法，将化合物4a替换为4d，将2-氟丙烯酸替换为2-溴丙烯酸，制备化合物5-12(197mg，产率26％，LC-ESI-MS：[M+H]＝581.1)。Synthesis procedure Reference Example 7. Compound 5a (197 mg, yield 26%, was prepared by a procedure similar to compound 5-1, substituting compound 4a for 4d and 2-fluoroacrylic acid for 2-bromoacrylic acid. LC-ESI-MS: [M+H] = 581.1.
1H-NMR(δ,CDCl3):8.37(s,1H,ArH),7.63(d,2H,J＝8.0Hz,ArH),7.05(d,2H,J＝8.0Hz,ArH),6.62(d,1H,J＝8.0Hz,ArH),6.40(d,1H,J＝8.0Hz,ArH),6.27(s,1H,ArH),6.08(s,2H,-NH2),5.27(d,0.5H,J＝4.0Hz,-C＝CH2),5.06(d,0.5H,J＝4.0Hz,-C＝CH2),5.03(d,1H,-C＝CH2),4.91(m,1H,piperidine),4.62(m,0.5H,piperidine),4.45(m,0.5H,piperidine),4.11(m,0.5H,piperidine),3.75(m,3.5H,-OCH3,piperidine),3.66(s,3H,-OCH3),3.43(m,0.5H,piperidine),3.12(m,0.5H,piperidine),2.87(m,0.5H,piperidine),2.76(m,0.5H,piperidine),2.27(m,2H,piperidine),1.75(m,2H,piperidine). 1 H-NMR (δ, CDCl 3 ): 8.37 (s, 1H, ArH), 7.63 (d, 2H, J = 8.0 Hz, ArH), 7.05 (d, 2H, J = 8.0 Hz, ArH), 6.62 ( d, 1H, J = 8.0 Hz, ArH), 6.40 (d, 1H, J = 8.0 Hz, ArH), 6.27 (s, 1H, ArH), 6.08 (s, 2H, -NH 2 ), 5.27 (d, 0.5H, J=4.0Hz, -C=CH 2 ), 5.06(d, 0.5H, J=4.0Hz, -C=CH 2 ), 5.03(d,1H,-C=CH 2 ), 4.91(m , 1H, piperidine), 4.62 (m, 0.5H, piperididine), 4.45 (m, 0.5H, piperididine), 4.11 (m, 0.5H, piperididine), 3.75 (m, 3.5H, -OCH 3 , piperididine), 3.66 (s, 3H, -OCH 3 ), 3.43 (m, 0.5H, piperididine), 3.12 (m, 0.5H, piperididine), 2.87 (m, 0.5H, piperididine), 2.76 (m, 0.5H, piperididine) , 2.27 (m, 2H, piperididine), 1.75 (m, 2H, piperididine).
实施例19. 1-(3-(4-氨基-3-(4-(4-氟苯氧基)苯基)-1H-吡唑并[3,4-d]嘧啶-1-基)哌啶-1-基)-2-氟丙基-2-烯-1-酮的制备(化合物5-13)Example 19. 1-(3-(4-Amino-3-(4-(4-fluorophenoxy)phenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin Preparation of pyridin-1-yl)-2-fluoropropyl-2-en-1-one (Compound 5-13)
Figure PCTCN2016081669-appb-000031
Figure PCTCN2016081669-appb-000031
合成步骤参考实施例7.用类似于化合物5-1的合成方法，将化合物4a替换为4e，制备化合物5-13(223mg，产率36％，LC-ESI-MS：[M+H]＝477.2)。Synthesis procedure Reference Example 7. Compound 5a (223 mg, yield 36%, LC-ESI-MS: [M+H] = 477.2).
1H-NMR(δ,CDCl3):8.35(s,1H,ArH),7.65(d,2H,J＝8.0Hz,ArH),7.47(d,2H,J＝8.0Hz,ArH),7.25(d,2H,J＝8.0Hz,ArH),7.02(d,2H,J＝8.0Hz,ArH),6.12(s,2H,-NH2),5.31(d,0.5H,J＝4.0Hz,-C＝CH2),5.18(d,0.5H,J＝4.0Hz,-C＝CH2),5.10(d,1H,-C＝CH2),4.92(m,1H,piperidine),4.66(m,0.5H,piperidine),4.44(m,0.5H,piperidine),4.12(m,0.5H,piperidine),3.75(m,0.5H,piperidine),3.44(m,0.5H,piperidine),3.19(m,0.5H,piperidine),2.87(m,0.5H,piperidine),2.75(m,0.5H,piperidine),2.30(m,2H,piperidine),1.77(m,2H,piperidine). 1 H-NMR (δ, CDCl 3 ): 8.35 (s, 1H, ArH), 7.65 (d, 2H, J = 8.0 Hz, ArH), 7.47 (d, 2H, J = 8.0 Hz, ArH), 7.25 ( d, 2H, J = 8.0 Hz, ArH), 7.02 (d, 2H, J = 8.0 Hz, ArH), 6.12 (s, 2H, -NH 2 ), 5.31 (d, 0.5H, J = 4.0 Hz, - C=CH 2 ), 5.18 (d, 0.5H, J=4.0 Hz, -C=CH 2 ), 5.10 (d, 1H, -C=CH 2 ), 4.92 (m, 1H, piperididine), 4.66 (m) , 0.5H, piperidine), 4.44 (m, 0.5H, piperididine), 4.12 (m, 0.5H, piperididine), 3.75 (m, 0.5H, piperididine), 3.44 (m, 0.5H, piperididine), 3.19 (m) , 0.5H, piperidine), 2.87 (m, 0.5H, piperididine), 2.75 (m, 0.5H, piperididine), 2.30 (m, 2H, piperididine), 1.77 (m, 2H, piperididine).
实施例20. 1-(3-(4-氨基-3-(4-(4-氟苯氧基)苯基)-1H-吡唑并[3,4-d]嘧啶-1-基)哌啶-1-基)-2-氯丙基-2-烯-1-酮的制备(化合物5-14)Example 20. 1-(3-(4-Amino-3-(4-(4-fluorophenoxy)phenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin Preparation of pyridin-1-yl)-2-chloropropyl-2-en-1-one (Compound 5-14)
Figure PCTCN2016081669-appb-000032
Figure PCTCN2016081669-appb-000032
合成步骤参考实施例7.用类似于化合物5-1的合成方法，将化合物4a替换为4e，将2-氟丙烯酸替换为2-氯丙烯酸，制备化合物5-14(254mg，产率40％，LC-ESI-MS：[M+H]＝493.1)。Synthesis procedure Reference Example 7. Compound 5a (254 mg, yield 40%, was prepared by a procedure similar to compound 5-1, substituting compound 4a to 4e and 2-fluoroacrylic acid to 2-chloroacrylic acid. LC-ESI-MS: [M+H] =495.
1H-NMR(δ,CDCl3-):8.37(s,1H,ArH),7.65(d,2H,J＝8.0Hz,ArH),7.47(d,2H,J＝8.0Hz,ArH),7.25(d,2H,J＝8.0Hz,ArH),7.02(d,2H,J＝8.0Hz,ArH),6.12(s,2H,-NH2),5.32(d,0.5H,J＝4.0Hz,-C＝CH2),5.19(d,0.5H,J＝4.0Hz,-C＝CH2),5.10(d,1H,-C＝CH2),4.92(m,1H,piperidine),4.65(m,0.5H,piperidine),4.44(m,0.5H,piperidine),4.12(m,0.5H,piperidine),3.75(m,0.5H,piperidine),3.44(m,0.5H,piperidine),3.19(m,0.5H,piperidine),2.89(m,0.5H,piperidine),2.77(m,0.5H,piperidine),2.32(m,2H,piperidine),1.76(m,2H,piperidine). 1 H-NMR (δ, CDCl 3 -): 8.37 (s, 1H, ArH), 7.65 (d, 2H, J = 8.0 Hz, ArH), 7.47 (d, 2H, J = 8.0 Hz, ArH), 7.25 (d, 2H, J = 8.0 Hz, ArH), 7.02 (d, 2H, J = 8.0 Hz, ArH), 6.12 (s, 2H, -NH 2 ), 5.32 (d, 0.5H, J = 4.0 Hz, -C=CH 2 ), 5.19 (d, 0.5H, J = 4.0 Hz, -C=CH 2 ), 5.10 (d, 1H, -C=CH 2 ), 4.92 (m, 1H, piperididine), 4.65 ( m, 0.5H, piperidine), 4.44 (m, 0.5H, piperididine), 4.12 (m, 0.5H, piperididine), 3.75 (m, 0.5H, piperididine), 3.44 (m, 0.5H, piperididine), 3.19 ( m, 0.5H, piperididine), 2.89 (m, 0.5H, piperididine), 2.77 (m, 0.5H, piperididine), 2.32 (m, 2H, piperididine), 1.76 (m, 2H, piperididine).
实施例21. 1-(3-(4-氨基-3-(4-(4-氟苯氧基)苯基)-1H-吡唑并[3,4-d]嘧啶-1-基)哌啶-1-基)-2-溴丙基-2-烯-1-酮的制备(化合物5-15) Example 21. 1-(3-(4-Amino-3-(4-(4-fluorophenoxy)phenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin Preparation of pyridin-1-yl)-2-bromopropyl-2-en-1-one (Compound 5-15)
Figure PCTCN2016081669-appb-000033
Figure PCTCN2016081669-appb-000033
合成步骤参考实施例7.用类似于化合物5-1的合成方法，将化合物4a替换为4e，将2-氟丙烯酸替换为2-溴丙烯酸，制备化合物5-15(213mg，产率30％，LC-ESI-MS：[M+H]＝538.1)。Synthesis procedure Reference Example 7. Compound 5a (213 mg, yield 30%, was prepared by a procedure similar to compound 5-1, substituting compound 4a for 4e and 2-fluoroacrylic acid for 2-bromoacrylic acid. LC-ESI-MS: [M+H] =353.
1H-NMR(δ,CDCl3):8.36(s,1H,ArH),7.65(d,2H,J＝8.0Hz,ArH),7.47(d,2H,J＝8.0Hz,ArH),7.24(d,2H,J＝8.0Hz,ArH),7.06(d,2H,J＝8.0Hz,ArH),6.09(s,2H,-NH2),5.31(d,0.5H,J＝4.0Hz,-C＝CH2),5.18(d,0.5H,J＝4.0Hz,-C＝CH2),5.10(d,1H,-C＝CH2),4.92(m,1H,piperidine),4.63(m,0.5H,piperidine),4.44(m,0.5H,piperidine),4.12(m,0.5H,piperidine),3.75(m,0.5H,piperidine),3.43(m,0.5H,piperidine),3.19(m,0.5H,piperidine),2.87(m,0.5H,piperidine),2.79(m,0.5H,piperidine),2.31(m,2H,piperidine),1.72(m,2H,piperidine). 1 H-NMR (δ, CDCl 3 ): 8.36 (s, 1H, ArH), 7.65 (d, 2H, J = 8.0 Hz, ArH), 7.47 (d, 2H, J = 8.0 Hz, ArH), 7.24 ( d, 2H, J = 8.0 Hz, ArH), 7.06 (d, 2H, J = 8.0 Hz, ArH), 6.09 (s, 2H, -NH 2 ), 5.31 (d, 0.5H, J = 4.0 Hz, - C=CH 2 ), 5.18 (d, 0.5H, J=4.0 Hz, -C=CH 2 ), 5.10 (d, 1H, -C=CH 2 ), 4.92 (m, 1H, piperididine), 4.63 (m) , 0.5H, piperidine), 4.44 (m, 0.5H, piperididine), 4.12 (m, 0.5H, piperididine), 3.75 (m, 0.5H, piperididine), 3.43 (m, 0.5H, piperididine), 3.19 (m) , 0.5H, piperidine), 2.87 (m, 0.5H, piperididine), 2.79 (m, 0.5H, piperididine), 2.31 (m, 2H, piperididine), 1.72 (m, 2H, piperididine).
实施例22. 1-(3-(4-氨基-3-(4-(4-氯苯氧基)苯基)-1H-吡唑并[3,4-d]嘧啶-1-基)哌啶-1-基)-2-氟丙基-2-烯-1-酮的制备(化合物5-16)Example 22. 1-(3-(4-Amino-3-(4-(4-chlorophenoxy)phenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin Preparation of pyridin-1-yl)-2-fluoropropyl-2-en-1-one (Compound 5-16)
Figure PCTCN2016081669-appb-000034
Figure PCTCN2016081669-appb-000034
合成步骤参考实施例7.用类似于化合物5-1的合成方法，将化合物4a替换为4f，制备化合物5-16(211mg，产率33％，LC-ESI-MS：[M+H]＝493.1)。Synthesis procedure Referring to Example 7. Compound 5a was prepared by substituting compound 4a to 4f using a synthetic procedure analogous to compound 5-1 (211 mg, yield 33%, LC-ESI-MS: [M+H] = 493.1).
1H-NMR(δ,CDCl3):8.37(s,1H,ArH),7.65(d,2H,J＝8.0Hz,ArH),7.43(d,2H,J＝8.0Hz,ArH),7.11(d,2H,J＝8.0Hz,ArH),6.89(d,2H,J＝8.0Hz,ArH),6.02(s,2H,-NH2),5.26(d,0.5H,J＝4.0Hz,-C＝CH2),5.13(d,0.5H,J＝4.0Hz,-C＝CH2),5.01(d,1H,-C＝CH2),4.92(m,1H,piperidine),4.66(m,0.5H,piperidine),4.44(m,0.5H,piperidine),4.11(m,0.5H,piperidine),3.73(m,0.5H,piperidine),3.44(m,0.5H,piperidine),3.19(m,0.5H,piperidine),2.86(m,0.5H,piperidine),2.73(m,0.5H,piperidine),2.30(m,2H,piperidine),1.74(m,2H,piperidine). 1 H-NMR (δ, CDCl 3 ): 8.37 (s, 1H, ArH), 7.65 (d, 2H, J = 8.0 Hz, ArH), 7.43 (d, 2H, J = 8.0 Hz, ArH), 7.11 ( d, 2H, J = 8.0 Hz, ArH), 6.89 (d, 2H, J = 8.0 Hz, ArH), 6.02 (s, 2H, -NH 2 ), 5.26 (d, 0.5H, J = 4.0 Hz, - C=CH 2 ), 5.13 (d, 0.5H, J=4.0 Hz, -C=CH 2 ), 5.01 (d, 1H, -C=CH 2 ), 4.92 (m, 1H, piperididine), 4.66 (m) , 0.5H, piperidine), 4.44 (m, 0.5H, piperididine), 4.11 (m, 0.5H, piperididine), 3.73 (m, 0.5H, piperididine), 3.44 (m, 0.5H, piperididine), 3.19 (m) , 0.5H, piperidine), 2.86 (m, 0.5H, piperididine), 2.73 (m, 0.5H, piperididine), 2.30 (m, 2H, piperididine), 1.74 (m, 2H, piperididine).
实施例23. 1-(3-(4-氨基-3-(4-(4-氯苯氧基)苯基)-1H-吡唑并[3,4-d]嘧啶-1-基)哌啶-1-基)-2-氯丙基-2-烯-1-酮的制备(化合物5-17) Example 23. 1-(3-(4-Amino-3-(4-(4-chlorophenoxy)phenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin Preparation of pyridin-1-yl)-2-chloropropyl-2-en-1-one (Compound 5-17)
Figure PCTCN2016081669-appb-000035
Figure PCTCN2016081669-appb-000035
合成步骤参考实施例7.用类似于化合物5-1的合成方法，将化合物4a替换为4f，将2-氟丙烯酸替换为2-氯丙烯酸，制备化合物5-17(245mg，产率37％，LC-ESI-MS：[M+H]＝509.1)。Synthesis procedure Referring to Example 7. Compound 5a (245 mg, yield 37%, was prepared by a procedure similar to compound 5-1, substituting compound 4a to 4f and 2-fluoroacrylic acid to 2-chloroacrylic acid. LC-ESI-MS: [M+H] = 509.1.
1H-NMR(δ,CDCl3):8.37(s,1H,ArH),7.65(d,2H,J＝8.0Hz,ArH),7.43(d,2H,J＝8.0Hz,ArH),7.11(d,2H,J＝8.0Hz,ArH),6.89(d,2H,J＝8.0Hz,ArH),6.02(s,2H,-NH2),5.19(d,0.5H,J＝4.0Hz,-C＝CH2),5.10(d,0.5H,J＝4.0Hz,-C＝CH2),5.00(d,1H,-C＝CH2),4.92(m,1H,piperidine),4.66(m,0.5H,piperidine),4.44(m,0.5H,piperidine),4.11(m,0.5H,piperidine),3.73(m,0.5H,piperidine),3.44(m,0.5H,piperidine),3.19(m,0.5H,piperidine),2.86(m,0.5H,piperidine),2.77(m,0.5H,piperidine),2.31(m,2H,piperidine),1.74(m,2H,piperidine). 1 H-NMR (δ, CDCl 3 ): 8.37 (s, 1H, ArH), 7.65 (d, 2H, J = 8.0 Hz, ArH), 7.43 (d, 2H, J = 8.0 Hz, ArH), 7.11 ( d, 2H, J = 8.0 Hz, ArH), 6.89 (d, 2H, J = 8.0 Hz, ArH), 6.02 (s, 2H, -NH 2 ), 5.19 (d, 0.5H, J = 4.0 Hz, - C=CH 2 ), 5.10 (d, 0.5H, J=4.0 Hz, -C=CH 2 ), 5.00 (d, 1H, -C=CH 2 ), 4.92 (m, 1H, piperididine), 4.66 (m) , 0.5H, piperidine), 4.44 (m, 0.5H, piperididine), 4.11 (m, 0.5H, piperididine), 3.73 (m, 0.5H, piperididine), 3.44 (m, 0.5H, piperididine), 3.19 (m) , 0.5H, piperidine), 2.86 (m, 0.5H, piperididine), 2.77 (m, 0.5H, piperididine), 2.31 (m, 2H, piperididine), 1.74 (m, 2H, piperididine).
实施例24. 1-(3-(4-氨基-3-(4-(4-氯苯氧基)苯基)-1H-吡唑并[3,4-d]嘧啶-1-基)哌啶-1-基)-2-溴丙基-2-烯-1-酮的制备(化合物5-18)Example 24. 1-(3-(4-Amino-3-(4-(4-chlorophenoxy)phenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin Preparation of pyridin-1-yl)-2-bromopropyl-2-en-1-one (Compound 5-18)
Figure PCTCN2016081669-appb-000036
Figure PCTCN2016081669-appb-000036
合成步骤参考实施例7.用类似于化合物5-1的合成方法，将化合物4a替换为4f，将2-氟丙烯酸替换为2-溴丙烯酸，制备化合物5-18(226mg，产率31％，LC-ESI-MS：[M+H]＝554.1)。Synthesis procedure Reference Example 7. Compound 5a (226 mg, yield 31%, was prepared by a procedure similar to compound 5-1, substituting compound 4a to 4f and 2-fluoroacrylic acid to 2-bromoacrylic acid. LC-ESI-MS: [M+H] = 554.1.
1H-NMR(δ,CDCl3):8.37(s,1H,ArH),7.65(d,2H,J＝8.0Hz,ArH),7.43(d,2H,J＝8.0Hz,ArH),7.11(d,2H,J＝8.0Hz,ArH),6.89(d,2H,J＝8.0Hz,ArH),6.12(s,2H,-NH2),5.19(d,0.5H,J＝4.0Hz,-C＝CH2),5.12(d,0.5H,J＝4.0Hz,-C＝CH2),5.00(d,1H,-C＝CH2),4.92(m,1H,piperidine),4.66(m,0.5H,piperidine),4.40(m,0.5H,piperidine),4.11(m,0.5H,piperidine),3.75(m,0.5H,piperidine),3.39(m,0.5H,piperidine),3.19(m,0.5H,piperidine),2.89(m,0.5H,piperidine),2.74(m,0.5H,piperidine),2.27(m,2H,piperidine),1.75(m,2H,piperidine). 1 H-NMR (δ, CDCl 3 ): 8.37 (s, 1H, ArH), 7.65 (d, 2H, J = 8.0 Hz, ArH), 7.43 (d, 2H, J = 8.0 Hz, ArH), 7.11 ( d, 2H, J = 8.0 Hz, ArH), 6.89 (d, 2H, J = 8.0 Hz, ArH), 6.12 (s, 2H, -NH 2 ), 5.19 (d, 0.5H, J = 4.0 Hz, - C=CH 2 ), 5.12 (d, 0.5H, J=4.0 Hz, -C=CH 2 ), 5.00 (d, 1H, -C=CH 2 ), 4.92 (m, 1H, piperididine), 4.66 (m) , 0.5H, piperidine), 4.40 (m, 0.5H, piperididine), 4.11 (m, 0.5H, piperididine), 3.75 (m, 0.5H, piperididine), 3.39 (m, 0.5H, piperididine), 3.19 (m) , 0.5H, piperidine), 2.89 (m, 0.5H, piperididine), 2.74 (m, 0.5H, piperididine), 2.27 (m, 2H, piperididine), 1.75 (m, 2H, piperididine).
实施例25.关键中间体4g的制备 Example 25. Preparation of key intermediate 4g
Figure PCTCN2016081669-appb-000037
Figure PCTCN2016081669-appb-000037
步骤1. 3-(6-苯氧基吡啶-3-基)-1H-吡唑并[3,4-d]嘧啶-4-胺(化合物2g)的合成Step 1. Synthesis of 3-(6-phenoxypyridin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (Compound 2g)
合成步骤参考实施例1步骤1.用类似于化合物2a的合成方法，从6-苯氧基吡啶-3-基硼酸(4.46g,18mmol)制备化合物2g(2.25g，产率67％)，LC-ESI-MS：[M+H]＝305.1。Synthesis procedure Reference Example 1 Step 1. Compound 2g (2.25 g, yield 67%) was prepared from 6-phenoxypyridin-3-ylboronic acid (4.46 g, 18 mmol) using a procedure similar to compound 2a. - ESI-MS: [M+H] = 305.1.
步骤2. 3-(4-氨基-3-(6-苯氧基吡啶-3-基)-1H-吡唑并[3,4-d]嘧啶-1-基)哌啶-1-甲酸叔丁酯(化合物3g)的合成Step 2. 3-(4-Amino-3-(6-phenoxypyridin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidine-1-carboxylic acid Synthesis of Butyl Ester (Compound 3g)
合成步骤参考实施例1步骤2.用类似于化合物3a的合成方法制备化合物3g(1.40g,产率40％)，LC-ESI-MS：[M+H]＝488.2。Synthesis procedure Reference Example 1 Step 2. Compound 3g (1.40 g, yield 40%) was obtained by the procedure of compound 3a, LC-ESI-MS: [M+H]=488.2.
步骤3. 3-(6-苯氧基吡啶-3-基)-1-(哌啶-3-基)-1H-吡唑并[3,4-d]嘧啶-4-胺盐酸盐(化合物4g)的合成Step 3. 3-(6-Phenoxypyridin-3-yl)-1-(piperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine hydrochloride ( Synthesis of compound 4g)
合成步骤参考实施例1步骤3.用类似于化合物4a的合成方法制备化合物4g(0.71g,产率80％，LC-ESI-MS：[M+H]＝388.2)。Synthesis procedure Reference Example 1 Step 3. Compound 4g (0.71 g, yield 80%, LC-ESI-MS: [M+H]= 388.2).
实施例26.关键中间体4h的制备Example 26. Preparation of key intermediate 4h
Figure PCTCN2016081669-appb-000038
Figure PCTCN2016081669-appb-000038
步骤1. 3-(6-(2-氟苯氧基)吡啶-3-基)-1H-吡唑并[3,4-d]嘧啶-4-胺(化合物2h)的合成Step 1. Synthesis of 3-(6-(2-fluorophenoxy)pyridin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (Compound 2h)
合成步骤参考实施例1步骤1.用类似于化合物2a的合成方法，从6-(2-氟苯氧基)吡啶-3-基硼酸(4.46g,18mmol)制备化合物2h(2.31g，产率68％)，LC-ESI-MS：[M+H]＝323.1。Synthesis procedure Reference Example 1 Step 1. Preparation of compound 2h (2.31 g, yield from 6-(2-fluorophenoxy)pyridin-3-ylboronic acid (4.46 g, 18 mmol) using a procedure similar to compound 2a 68%), LC-ESI-MS: [M+H]=323.1.
步骤2. 3-(4-氨基-3-(6-(2-氟苯氧基)吡啶-3-基)-1H-吡唑并[3,4-d]嘧啶-1-基)哌啶-1-甲酸叔丁酯(化合物3h)的合成Step 2. 3-(4-Amino-3-(6-(2-fluorophenoxy)pyridin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidine Synthesis of 1-butylic acid tert-butyl ester (compound 3h)
合成步骤参考实施例1步骤2.用类似于化合物3a的合成方法制备化合物3h(1.40g,产率40％)，LC-ESI-MS：[M+H]＝506.1。Synthesis procedure Reference Example 1 Step 2. Compound 3h (1.40 g, yield 40%) was obtained by the procedure of compound 3a, LC-ESI-MS: [M+H]=506.1.
步骤3. 3-(6-(2-氟苯氧基)吡啶-3-基)-1-(哌啶-3-基)-1H-吡唑并[3,4-d]嘧啶-4-胺盐酸盐(化合物4h)的合成Step 3. 3-(6-(2-Fluorophenoxy)pyridin-3-yl)-1-(piperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-4- Synthesis of Amine Hydrochloride (Compound 4h)
合成步骤参考实施例1步骤3.用类似于化合物4a的合成方法制备化合物4h(0.67g,产率77％，LC-ESI-MS：[M+H]＝406.2)。 Synthesis procedure Reference Example 1 Step 3. Compound 4h (0.67 g, yield 77%, LC-ESI-MS: [M+H] = 406.2).
实施例27.关键中间体4i的制备Example 27. Preparation of key intermediate 4i
Figure PCTCN2016081669-appb-000039
Figure PCTCN2016081669-appb-000039
步骤1. 3-(6-(4-氟苯氧基)吡啶-3-基)-1H-吡唑并[3,4-d]嘧啶-4-胺(化合物2i)的合成Step 1. Synthesis of 3-(6-(4-fluorophenoxy)pyridin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (Compound 2i)
合成步骤参考实施例1步骤1.用类似于化合物2a的合成方法，从6-(4-氟苯氧基)吡啶-3-基硼酸(4.46g,18mmol)制备化合物2i(2.32g，产率68％)，LC-ESI-MS：[M+H]＝323.1。Synthesis procedure Reference Example 1 Step 1. Preparation of compound 2i (2.32 g, yield from 6-(4-fluorophenoxy)pyridin-3-ylboronic acid (4.46 g, 18 mmol) using a procedure similar to compound 2a 68%), LC-ESI-MS: [M+H]=323.1.
步骤2. 3-(4-氨基-3-(6-(4-氟苯氧基)吡啶-3-基)-1H-吡唑并[3,4-d]嘧啶-1-基)哌啶-1-甲酸叔丁酯(化合物3i)的合成Step 2. 3-(4-Amino-3-(6-(4-fluorophenoxy)pyridin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidine Synthesis of tert-butyl-1-carboxylic acid (compound 3i)
合成步骤参考实施例1步骤2.用类似于化合物3a的合成方法制备化合物3i(1.32g,产率38％)，LC-ESI-MS：[M+H]＝506.2。Synthesis procedure Reference Example 1 Step 2. Compound 3i (1.32 g, yield 38%) was obtained using a procedure similar to compound 3a, LC-ESI-MS: [M+H]=506.2.
步骤3. 3-(6-(4-氟苯氧基)吡啶-3-基)-1-(哌啶-3-基)-1H-吡唑并[3,4-d]嘧啶-4-胺盐酸盐(化合物4i)的合成Step 3. 3-(6-(4-Fluorophenoxy)pyridin-3-yl)-1-(piperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-4- Synthesis of Amine Hydrochloride (Compound 4i)
合成步骤参考实施例1步骤3.用类似于化合物4a的合成方法制备化合物4i(0.68g,产率78％，LC-ESI-MS：[M+H]＝406.2)。Synthesis procedure Reference Example 1 Step 3. Compound 4i (0.68 g, yield 78%, LC-ESI-MS: [M+H] = 406.2) was obtained.
实施例28. 1-(3-(4-氨基-3-(6-苯氧基吡啶-3-基)-1H-吡唑并[3,4-d]嘧啶-1-基)哌啶-1-基)-2-溴丙基-2-烯-1-酮的制备(化合物5-19)Example 28. 1-(3-(4-Amino-3-(6-phenoxypyridin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidine- Preparation of 1-yl)-2-bromopropyl-2-en-1-one (Compound 5-19)
Figure PCTCN2016081669-appb-000040
Figure PCTCN2016081669-appb-000040
合成步骤参考实施例7.用类似于化合物5-1的合成方法，将化合物4a替换为4g，将2-氟丙烯酸替换为2-溴丙烯酸，制备化合物5-19(202mg，产率30％，LC-ESI-MS：[M+H]＝520.1)。Synthesis procedure Reference Example 7. Compound 5a (202 mg, yield 30%, was prepared by a procedure similar to compound 5-1, substituting compound 4a to 4 g and 2-fluoroacrylic acid to 2-bromoacrylic acid. LC-ESI-MS: [M+H] = 520.1.
1H-NMR(δ,CDCl3):8.50(d,1H,J＝2.0Hz,ArH),8.35(s,1H,ArH),8.03(dd,1H,J1＝8.4Hz,J2＝2.4Hz,ArH),7.42(td,2H,J1＝7.6Hz,J2＝2.0Hz,ArH),7.23(t,1H,J＝7.6Hz,ArH),7.18(dd,2H,J1＝8.4Hz,J2＝2.0Hz,ArH),7.08(d,1H,J＝8.4Hz,ArH),6.04(s,2H,-NH2),5.19(d,0.5H,J＝4.0Hz,-C＝CH2),5.10(d,0.5H,J＝4.0Hz,-C＝CH2),5.05(d,1H,-C＝CH2),4.91(m,1H,piperidine),4.63(m,0.5H,piperidine),4.45(m,0.5H,piperidine),4.12(m,0.5H,piperidine),3.75(m,0.5H,piperidine),3.43(m,0.5H,piperidine),3.17(m,0.5H,piperidine),2.85(m,0.5H, piperidine),2.77(m,0.5H,piperidine),2.30(m,2H,piperidine),1.73(m,2H,piperidine). 1 H-NMR (δ, CDCl 3 ): 8.50 (d, 1H, J = 2.0 Hz, ArH), 8.35 (s, 1H, ArH), 8.03 (dd, 1H, J 1 = 8.4 Hz, J 2 = 2.4 Hz, ArH), 7.42 (td, 2H, J 1 = 7.6 Hz, J 2 = 2.0 Hz, ArH), 7.23 (t, 1H, J = 7.6 Hz, ArH), 7.18 (dd, 2H, J 1 = 8.4 Hz, J 2 = 2.0 Hz, ArH), 7.08 (d, 1H, J = 8.4 Hz, ArH), 6.04 (s, 2H, -NH 2 ), 5.19 (d, 0.5H, J = 4.0 Hz, -C =CH 2 ), 5.10 (d, 0.5H, J = 4.0 Hz, -C=CH 2 ), 5.05 (d, 1H, -C=CH 2 ), 4.91 (m, 1H, piperididine), 4.63 (m, 0.5H,piperidine), 4.45 (m, 0.5H, piperididine), 4.12 (m, 0.5H, piperididine), 3.75 (m, 0.5H, piperididine), 3.43 (m, 0.5H, piperididine), 3.17 (m, 0.5H, piperidine), 2.85 (m, 0.5H, piperidine), 2.77 (m, 0.5H, piperididine), 2.30 (m, 2H, piperididine), 1.73 (m, 2H, piperididine).
实施例29. 1-(3-(4-氨基-3-(6-苯氧基吡啶-3-基)-1H-吡唑并[3,4-d]嘧啶-1-基)哌啶-1-基)-2-氯丙基-2-烯-1-酮的制备(化合物5-20)Example 29. 1-(3-(4-Amino-3-(6-phenoxypyridin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidine- Preparation of 1-yl)-2-chloropropyl-2-en-1-one (Compound 5-20)
Figure PCTCN2016081669-appb-000041
Figure PCTCN2016081669-appb-000041
合成步骤参考实施例7.用类似于化合物5-1的合成方法，将化合物4a替换为4g，将2-氟丙烯酸替换为2-氯丙烯酸，制备化合物5-20(214mg，产率35％，LC-ESI-MS：[M+H]＝476.2)。Synthesis procedure Reference Example 7. Compound 5a (214 mg, yield 35%, was prepared by a procedure similar to compound 5-1, substituting compound 4a to 4 g and 2-fluoroacrylic acid to 2-chloroacrylic acid. LC-ESI-MS: [M+H] = 476.2.
1H-NMR(δ,CDCl3):8.51(d,1H,J＝2.0Hz,ArH),8.36(s,1H,ArH),8.05(dd,1H,J1＝8.0Hz,J2＝2.0Hz,ArH),7.44(td,2H,J1＝7.6Hz,J2＝2.0Hz,ArH),7.24(t,1H,J＝7.6Hz,ArH),7.20(dd,2H,J1＝8.4Hz,J2＝2.0Hz,ArH),7.11(d,1H,J＝8.4Hz,ArH),5.98(s,2H,-NH2),5.21(d,0.5H,J＝4.0Hz,-C＝CH2),5.11(d,0.5H,J＝4.0Hz,-C＝CH2),5.06(d,1H,-C＝CH2),4.92(m,1H,piperidine),4.63(m,0.5H,piperidine),4.43(m,0.5H,piperidine),4.11(m,0.5H,piperidine),3.74(m,0.5H,piperidine),3.42(m,0.5H,piperidine),3.17(m,0.5H,piperidine),2.82(m,0.5H,piperidine),2.75(m,0.5H,piperidine),2.31(m,2H,piperidine),1.73(m,2H,piperidine). 1 H-NMR (δ, CDCl 3 ): 8.51 (d, 1H, J = 2.0 Hz, ArH), 8.36 (s, 1H, ArH), 8.05 (dd, 1H, J 1 = 8.0 Hz, J 2 = 2.0 Hz, ArH), 7.44 (td, 2H, J 1 = 7.6 Hz, J 2 = 2.0 Hz, ArH), 7.24 (t, 1H, J = 7.6 Hz, ArH), 7.20 (dd, 2H, J 1 = 8.4 Hz, J 2 = 2.0 Hz, ArH), 7.11 (d, 1H, J = 8.4 Hz, ArH), 5.98 (s, 2H, -NH 2 ), 5.21 (d, 0.5H, J = 4.0 Hz, -C =CH 2 ), 5.11 (d, 0.5H, J = 4.0 Hz, -C=CH 2 ), 5.06 (d, 1H, -C=CH 2 ), 4.92 (m, 1H, piperididine), 4.63 (m, 0.5H,piperidine), 4.43 (m, 0.5H, piperididine), 4.11 (m, 0.5H, piperididine), 3.74 (m, 0.5H, piperididine), 3.42 (m, 0.5H, piperididine), 3.17 (m, 0.5H,piperidine), 2.82 (m, 0.5H, piperididine), 2.75 (m, 0.5H, piperididine), 2.31 (m, 2H, piperididine), 1.73 (m, 2H, piperididine).
实施例30. 1-(3-(4-氨基-3-(6-(2-氟苯氧基)吡啶-3-基)-1H-吡唑并[3,4-d]嘧啶-1-基)哌啶-1-基)-2-溴丙基-2-烯-1-酮的制备(化合物5-21)Example 30. 1-(3-(4-Amino-3-(6-(2-fluorophenoxy)pyridin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-1- Preparation of phenyl)piperidin-1-yl)-2-bromopropyl-2-en-1-one (Compound 5-21)
Figure PCTCN2016081669-appb-000042
Figure PCTCN2016081669-appb-000042
合成步骤参考实施例7.用类似于化合物5-1的合成方法，将化合物4a替换为4h，将2-氟丙烯酸替换为2-溴丙烯酸，制备化合物5-21(227mg，产率29％，LC-ESI-MS：[M+H]＝538.1)。Synthesis procedure Reference Example 7. Compound 5-21 (227 mg, yield 29%, was prepared by a procedure similar to compound 5-1, substituting compound 4a for 4h and 2-fluoroacrylic acid for 2-bromoacrylic acid. LC-ESI-MS: [M+H] =353.
1H-NMR(δ,CDCl3):8.51(d,1H,J＝2.0Hz,ArH),8.39(s,1H,ArH),8.06(dd,1H,J1＝8.4Hz,J2＝2.0Hz,ArH),7.37(dd,2H,J1＝8.4Hz,J2＝7.2Hz,ArH),7.17(d,1H,J＝8.4Hz,ArH),6.94(m,2H,ArH),6.00(s,2H,-NH2),5.21(d,0.5H,J＝4.0Hz,-C＝CH2),5.11(d,0.5H,J＝4.0Hz,-C＝CH2),5.03(d,1H,-C＝CH2),4.92(m,1H,piperidine),4.60(m,0.5H,piperidine),4.42(m,0.5H,piperidine),4.11(m,0.5H,piperidine),3.72(m,0.5H,piperidine),3.43(m,0.5H,piperidine),3.13 (m,0.5H,piperidine),2.82(m,0.5H,piperidine),2.70(m,0.5H,piperidine),2.31(m,2H,piperidine),1.75(m,2H,piperidine). 1 H-NMR (δ, CDCl 3 ): 8.51 (d, 1H, J = 2.0 Hz, ArH), 8.39 (s, 1H, ArH), 8.06 (dd, 1H, J 1 = 8.4 Hz, J 2 = 2.0 Hz, ArH), 7.37 (dd, 2H, J 1 = 8.4 Hz, J 2 = 7.2 Hz, ArH), 7.17 (d, 1H, J = 8.4 Hz, ArH), 6.94 (m, 2H, ArH), 6.00 (s, 2H, -NH 2 ), 5.21 (d, 0.5H, J = 4.0 Hz, -C=CH 2 ), 5.11 (d, 0.5H, J = 4.0 Hz, -C=CH 2 ), 5.03 ( d, 1H, -C=CH 2 ), 4.92 (m, 1H, piperididine), 4.60 (m, 0.5H, piperididine), 4.42 (m, 0.5H, piperididine), 4.11 (m, 0.5H, piperididine), 3.72 (m, 0.5H, piperidine), 3.43 (m, 0.5H, piperididine), 3.13 (m, 0.5H, piperididine), 2.82 (m, 0.5H, piperididine), 2.70 (m, 0.5H, piperididine), 2.31 (m, 2H, piperididine), 1.75 (m, 2H, piperididine).
实施例31. 1-(3-(4-氨基-3-(6-(2-氟苯氧基)吡啶-3-基)-1H-吡唑并[3,4-d]嘧啶-1-基)哌啶-1-基)-2-氯丙基-2-烯-1-酮的制备(化合物5-22)Example 31. 1-(3-(4-Amino-3-(6-(2-fluorophenoxy)pyridin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-1- Preparation of phenyl)piperidin-1-yl)-2-chloropropyl-2-en-1-one (Compound 5-22)
Figure PCTCN2016081669-appb-000043
Figure PCTCN2016081669-appb-000043
合成步骤参考实施例7.用类似于化合物5-1的合成方法，将化合物4a替换为4h，将2-氟丙烯酸替换为2-氯丙烯酸，制备化合物5-22(201mg，产率31％，LC-ESI-MS：[M+H]＝494.1)。Synthesis procedure Reference Example 7. Compound 5a (201 mg, yield 31%) was prepared by a procedure similar to compound 5-1, substituting compound 4a for 4h and 2-fluoroacrylic acid for 2-chloroacrylic acid. LC-ESI-MS: [M+H] =495.
1H-NMR(δ,CDCl3):8.52(d,1H,J＝2.0Hz,ArH),8.39(s,1H,ArH),8.05(dd,1H,J1＝8.4Hz,J2＝2.0Hz,ArH),7.37(dd,2H,J1＝8.4Hz,J2＝7.6Hz,ArH),7.15(d,1H,J＝8.4Hz,ArH),6.94(m,2H,ArH),5.92(s,2H,-NH2),5.21(d,0.5H,J＝4.0Hz,-C＝CH2),5.12(d,0.5H,J＝4.0Hz,-C＝CH2),5.06(d,1H,-C＝CH2),4.91(m,1H,piperidine),4.63(m,0.5H,piperidine),4.41(m,0.5H,piperidine),4.11(m,0.5H,piperidine),3.75(m,0.5H,piperidine),3.44(m,0.5H,piperidine),3.16(m,0.5H,piperidine),2.81(m,0.5H,piperidine),2.75(m,0.5H,piperidine),2.32(m,2H,piperidine),1.74(m,2H,piperidine). 1 H-NMR (δ, CDCl 3 ): 8.52 (d, 1H, J = 2.0 Hz, ArH), 8.39 (s, 1H, ArH), 8.05 (dd, 1H, J 1 = 8.4 Hz, J 2 = 2.0 Hz, ArH), 7.37 (dd, 2H, J 1 = 8.4 Hz, J 2 = 7.6 Hz, ArH), 7.15 (d, 1H, J = 8.4 Hz, ArH), 6.94 (m, 2H, ArH), 5.92 (s, 2H, -NH 2 ), 5.21 (d, 0.5H, J = 4.0 Hz, -C=CH 2 ), 5.12 (d, 0.5H, J = 4.0 Hz, -C=CH 2 ), 5.06 ( d, 1H, -C=CH 2 ), 4.91 (m, 1H, piperididine), 4.63 (m, 0.5H, piperididine), 4.41 (m, 0.5H, piperididine), 4.11 (m, 0.5H, piperididine), 3.75 (m, 0.5H, piperidine), 3.44 (m, 0.5H, piperididine), 3.16 (m, 0.5H, piperididine), 2.81 (m, 0.5H, piperididine), 2.75 (m, 0.5H, piperididine), 2.32 (m, 2H, piperididine), 1.74 (m, 2H, piperididine).
实施例32. 1-(3-(4-氨基-3-(6-(4-氟苯氧基)吡啶-3-基)-1H-吡唑并[3,4-d]嘧啶-1-基)哌啶-1-基)-2-溴丙基-2-烯-1-酮的制备(化合物5-23)Example 32. 1-(3-(4-Amino-3-(6-(4-fluorophenoxy)pyridin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-1- Preparation of phenyl)piperidin-1-yl)-2-bromopropyl-2-en-1-one (Compound 5-23)
Figure PCTCN2016081669-appb-000044
Figure PCTCN2016081669-appb-000044
合成步骤参考实施例7.用类似于化合物5-1的合成方法，将化合物4a替换为4i，将2-氟丙烯酸替换为2-溴丙烯酸，制备化合物5-23(234mg，产率30％，LC-ESI-MS：[M+H]＝538.1)。Synthesis procedure Referring to Example 7. Compound 5a (234 mg, yield 30%, was prepared by a procedure similar to compound 5-1, substituting compound 4a to 4i and 2-fluoroacrylic acid to 2-bromoacrylic acid. LC-ESI-MS: [M+H] =353.
1H-NMR(δ,CDCl3):8.41(d,1H,J＝2.0Hz,ArH),8.28(s,1H,ArH),7.96(dd,1H,J1＝8.8Hz,J2＝2.4Hz,ArH),7.01(m,5H,ArH),5.84(s,2H,-NH2),5.29(d,0.5H,J＝4.0Hz,-C＝CH2),5.13(d,0.5H,J＝4.0Hz,-C＝CH2),5.03(d,1H,-C＝CH2),4.92(m,1H,piperidine),4.61(m,0.5H, piperidine),4.44(m,0.5H,piperidine),4.12(m,0.5H,piperidine),3.74(m,0.5H,piperidine),3.41(m,0.5H,piperidine),3.13(m,0.5H,piperidine),2.82(m,0.5H,piperidine),2.68(m,0.5H,piperidine),2.32(m,2H,piperidine),1.75(m,2H,piperidine). 1 H-NMR (δ, CDCl 3 ): 8.41 (d, 1H, J = 2.0 Hz, ArH), 8.28 (s, 1H, ArH), 7.96 (dd, 1H, J 1 = 8.8 Hz, J 2 = 2.4 Hz, ArH), 7.01 (m, 5H, ArH), 5.84 (s, 2H, -NH 2 ), 5.29 (d, 0.5H, J = 4.0 Hz, -C = CH 2 ), 5.13 (d, 0.5H) , J=4.0 Hz, -C=CH 2 ), 5.03 (d, 1H, -C=CH 2 ), 4.92 (m, 1H, piperididine), 4.61 (m, 0.5H, piperidine), 4.44 (m, 0.5) H,piperidine), 4.12 (m, 0.5H, piperididine), 3.74 (m, 0.5H, piperididine), 3.41 (m, 0.5H, piperididine), 3.13 (m, 0.5H, piperididine), 2.82 (m, 0.5) H,piperidine), 2.68 (m, 0.5H, piperididine), 2.32 (m, 2H, piperididine), 1.75 (m, 2H, piperididine).
实施例33. 1-(3-(4-氨基-3-(6-(4-氟苯氧基)吡啶-3-基)-1H-吡唑并[3,4-d]嘧啶-1-基)哌啶-1-基)-2-氯丙基-2-烯-1-酮的制备(化合物5-24)Example 33. 1-(3-(4-Amino-3-(6-(4-fluorophenoxy)pyridin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-1- Preparation of phenyl)piperidin-1-yl)-2-chloropropyl-2-en-1-one (Compound 5-24)
Figure PCTCN2016081669-appb-000045
Figure PCTCN2016081669-appb-000045
合成步骤参考实施例7.用类似于化合物5-1的合成方法，将化合物4a替换为4i，将2-氟丙烯酸替换为2-氯丙烯酸，制备化合物5-24(205mg，产率31％，LC-ESI-MS：[M+H]＝494.1)。Synthesis procedure Reference Example 7. Compound 5a (205 mg, yield 31%, was prepared by a procedure similar to compound 5-1, substituting compound 4a to 4i and 2-fluoroacrylic acid to 2-chloroacrylic acid. LC-ESI-MS: [M+H] =495.
1H-NMR(δ,CDCl3):8.41(d,1H,J＝2.0Hz,ArH),8.28(s,1H,ArH),7.97(dd,1H,J1＝8.4Hz,J2＝2.0Hz,ArH),7.01(m,5H,ArH),5.92(s,2H,-NH2),5.21(d,0.5H,J＝4.0Hz,-C＝CH2),5.12(d,0.5H,J＝4.0Hz,-C＝CH2),5.06(d,1H,-C＝CH2),4.99(m,1H,piperidine),4.65(m,0.5H,piperidine),4.41(m,0.5H,piperidine),4.17(m,0.5H,piperidine),3.75(m,0.5H,piperidine),3.43(m,0.5H,piperidine),3.16(m,0.5H,piperidine),2.83(m,0.5H,piperidine),2.71(m,0.5H,piperidine),2.33(m,2H,piperidine),1.74(m,2H,piperidine). 1 H-NMR (δ, CDCl 3 ): 8.41 (d, 1H, J = 2.0 Hz, ArH), 8.28 (s, 1H, ArH), 7.97 (dd, 1H, J 1 = 8.4 Hz, J 2 = 2.0 Hz, ArH), 7.01 (m, 5H, ArH), 5.92 (s, 2H, -NH 2 ), 5.21 (d, 0.5H, J = 4.0 Hz, -C=CH 2 ), 5.12 (d, 0.5H) , J=4.0 Hz, -C=CH 2 ), 5.06 (d, 1H, -C=CH 2 ), 4.99 (m, 1H, piperididine), 4.65 (m, 0.5H, piperididine), 4.41 (m, 0.5) H,piperidine), 4.17 (m, 0.5H, piperididine), 3.75 (m, 0.5H, piperididine), 3.43 (m, 0.5H, piperididine), 3.16 (m, 0.5H, piperididine), 2.83 (m, 0.5) H,piperidine), 2.71 (m, 0.5H, piperididine), 2.33 (m, 2H, piperididine), 1.74 (m, 2H, piperididine).
实施例34. 1-(3-(4-氨基-3-(6-苯氧基吡啶-3-基)-1H-吡唑并[3,4-d]嘧啶-1-基)哌啶-1-基)-2-氟丙基-2-烯-1-酮的制备(化合物5-25)Example 34. 1-(3-(4-Amino-3-(6-phenoxypyridin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidine- Preparation of 1-yl)-2-fluoropropyl-2-en-1-one (Compound 5-25)
Figure PCTCN2016081669-appb-000046
Figure PCTCN2016081669-appb-000046
合成步骤参考实施例7.用类似于化合物5-1的合成方法，将化合物4a替换为4g，制备化合物5-25(195mg，产率33％，LC-ESI-MS：[M+H]＝460.2)。Synthesis procedure Reference Example 7. Compound 4a was replaced with 4 g by a synthetic method similar to Compound 5-1 to prepare Compound 5-25 (195 mg, yield 33%, LC-ESI-MS: [M+H] = 460.2).
1H-NMR(δ,CDCl3):8.51(d,1H,J＝2.0Hz,ArH),8.36(s,1H,ArH),8.05(dd,1H,J1＝8.0Hz,J2＝2.4Hz,ArH),7.43(td,2H,J1＝7.6Hz,J2＝2.0Hz,ArH),7.26(t,1H,J＝8.0Hz,ArH),7.21(dd,2H,J1＝8.4Hz,J2＝2.0Hz,ArH),7.12(d,1H,J＝8.4Hz,ArH),5.99(s,2H,-NH2),5.21(d, 0.5H,J＝4.0Hz,-C＝CH2),5.11(d,0.5H,J＝4.0Hz,-C＝CH2),5.06(d,1H,-C＝CH2),4.92(m,1H,piperidine),4.65(m,0.5H,piperidine),4.39(m,0.5H,piperidine),4.12(m,0.5H,piperidine),3.72(m,0.5H,piperidine),3.42(m,0.5H,piperidine),3.17(m,0.5H,piperidine),2.83(m,0.5H,piperidine),2.76(m,0.5H,piperidine),2.32(m,2H,piperidine),1.77(m,2H,piperidine). 1 H-NMR (δ, CDCl 3 ): 8.51 (d, 1H, J = 2.0 Hz, ArH), 8.36 (s, 1H, ArH), 8.05 (dd, 1H, J 1 = 8.0 Hz, J 2 = 2.4 Hz, ArH), 7.43 (td, 2H, J 1 = 7.6 Hz, J 2 = 2.0 Hz, ArH), 7.26 (t, 1H, J = 8.0 Hz, ArH), 7.21 (dd, 2H, J 1 = 8.4 Hz, J 2 = 2.0 Hz, ArH), 7.12 (d, 1H, J = 8.4 Hz, ArH), 5.99 (s, 2H, -NH 2 ), 5.21 (d, 0.5H, J = 4.0 Hz, -C =CH 2 ), 5.11 (d, 0.5H, J = 4.0 Hz, -C=CH 2 ), 5.06 (d, 1H, -C=CH 2 ), 4.92 (m, 1H, piperididine), 4.65 (m, 0.5H,piperidine), 4.39 (m,0.5H,piperidine), 4.12 (m,0.5H,piperidine), 3.72 (m,0.5H,piperidine), 3.42 (m,0.5H,piperidine), 3.17 (m, 0.5H,piperidine), 2.83 (m, 0.5H, piperididine), 2.76 (m, 0.5H, piperididine), 2.32 (m, 2H, piperididine), 1.77 (m, 2H, piperididine).
实施例35.手性关键中间体(S)-4a的制备Example 35. Preparation of Chiral Key Intermediate (S)-4a
Figure PCTCN2016081669-appb-000047
Figure PCTCN2016081669-appb-000047
步骤1.(S)-3-(4-氨基-3-(6-苯氧基吡啶-3-基)-1H-吡唑并[3,4-d]嘧啶-1-基)哌啶-1-甲酸叔丁酯(化合物(S)-3a)的合成Step 1. (S)-3-(4-Amino-3-(6-phenoxypyridin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidine- Synthesis of tert-butyl 1-carboxylate (compound (S)-3a)
合成步骤参考实施例1步骤2.以手性纯的(S)-3-羟基N-Boc哌啶为原料，用类似于化合物3a的合成方法制备化合物(S)-3a(产率51％)，LC-ESI-MS：[M+H]＝487.1。Synthesis procedure Referring to Example 1 Step 2. Using chiral pure (S)-3-hydroxy N-Boc piperidine as starting material, compound (S)-3a was prepared by a synthesis method similar to Compound 3a (yield 51%) LC-ESI-MS: [M+H] = 487.1.
步骤2.(S)-3-(6-苯氧基吡啶-3-基)-1-(哌啶-3-基)-1H-吡唑并[3,4-d]嘧啶-4-胺盐酸盐(化合物(S)-4a)的合成Step 2. (S)-3-(6-Phenoxypyridin-3-yl)-1-(piperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine Synthesis of hydrochloride (compound (S)-4a)
合成步骤参考实施例1步骤3.用类似于化合物4a的合成方法制备化合物(S)-4a(产率75％，LC-ESI-MS：[M+H]＝387.3)。Synthesis procedure Reference Example 1 Step 3. Compound (S)-4a was prepared by a synthetic method analogous to Compound 4a (yield: 75%, LC-ESI-MS: [M+H] = 387.3).
实施例36.手性关键中间体(S)-4g的制备Example 36. Preparation of Chiral Key Intermediate (S)-4g
Figure PCTCN2016081669-appb-000048
Figure PCTCN2016081669-appb-000048
步骤1.(S)-3-(4-氨基-3-(6-苯氧基吡啶-3-基)-1H-吡唑并[3,4-d]嘧啶-1-基)哌啶-1-甲酸叔丁酯(化合物(S)-3g)的合成Step 1. (S)-3-(4-Amino-3-(6-phenoxypyridin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidine- Synthesis of tert-butyl 1-carboxylate (compound (S)-3g)
合成步骤参考实施例1步骤2.以手性纯的(S)-3-羟基N-Boc哌啶为原料，用类似于化合物3a的合成方法制备化合物(S)-3g(1.40g,产率40％)，LC-ESI-MS：[M+H]＝488.2。The synthesis procedure is as follows. Step 2 of Example 2. Preparation of compound (S)-3g (1.40 g, yield by a synthesis method similar to compound 3a) using chiral pure (S)-3-hydroxy N-Boc piperidine as starting material. 40%), LC-ESI-MS: [M+H] = 488.2.
步骤2. 3-(6-苯氧基吡啶-3-基)-1-(哌啶-3-基)-1H-吡唑并[3,4-d]嘧啶-4-胺盐酸盐(化合物(S)-4g)的合成Step 2. 3-(6-Phenoxypyridin-3-yl)-1-(piperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine hydrochloride ( Synthesis of Compound (S)-4g)
合成步骤参考实施例1步骤3.用类似于化合物4a的合成方法制备化合物(S)-4g(0.71g,产率80％，LC-ESI-MS：[M+H]＝388.2)。 Synthesis procedure Reference Example 1 Step 3. Compound (S)-4g (0.71 g, yield 80%, LC-ESI-MS: [M+H]= 388.2) was prepared by a procedure similar to compound 4a.
实施例37(S)-1-(3-(4-氨基-3-(4-苯氧基苯基)-1H-吡唑并[3,4-d]嘧啶-1-基)哌啶-1-基)-2-氯丙基-2-烯-1-酮的制备(化合物(S)-5-2)Example 37(S)-1-(3-(4-Amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidine- Preparation of 1-yl)-2-chloropropyl-2-en-1-one (Compound (S)-5-2)
Figure PCTCN2016081669-appb-000049
Figure PCTCN2016081669-appb-000049
合成步骤参考实施例7.用类似于化合物5-1的合成方法，将2-氟丙烯酸替换为2-氯丙烯酸制备化合物5-2(312mg，产率51％，LC-ESI-MS：[M+H]＝475.2)。Synthesis procedure Reference Example 7. Compound 5-2 (312 mg, yield 51%, LC-ESI-MS: [M. +H]=475.2).
1H-NMR(δ,CDCl3):8.36(s,1H,ArH),7.64(d,2H,J＝8.0Hz,ArH),7.41(m,2H,ArH),7.16(m,3H,ArH),7.10(d,2H,J＝7.6Hz,ArH),5.66(s,2H,-NH2),5.61(m,2H,-C＝CH2),4.92(m,1H,piperidine),4.70(m,0.5H,piperidine),4.52(m,0.5H,piperidine),4.15(m,0.5H,piperidine),3.99(m,0.5H,piperidine),3.82(m,0.5H,piperidine),3.51(m,0.5H,piperidine),3.24(m,0.5H,piperidine),2.92(m,0.5H,piperidine),2.33(m,2H,piperidine),1.79(m,2H,piperidine). 1 H-NMR (δ, CDCl 3 ): 8.36 (s, 1H, ArH), 7.64 (d, 2H, J = 8.0 Hz, ArH), 7.41 (m, 2H, ArH), 7.16 (m, 3H, ArH) ), 7.10 (d, 2H, J = 7.6 Hz, ArH), 5.66 (s, 2H, -NH 2 ), 5.61 (m, 2H, -C=CH 2 ), 4.92 (m, 1H, piperididine), 4.70 (m, 0.5H, piperidine), 4.52 (m, 0.5H, piperididine), 4.15 (m, 0.5H, piperididine), 3.99 (m, 0.5H, piperididine), 3.82 (m, 0.5H, piperididine), 3.51 (m, 0.5H, piperidine), 3.24 (m, 0.5H, piperididine), 2.92 (m, 0.5H, piperididine), 2.33 (m, 2H, piperididine), 1.79 (m, 2H, piperididine).
实施例38.(S)-1-(3-(4-氨基-3-(4-苯氧基苯基)-1H-吡唑并[3,4-d]嘧啶-1-基)哌啶-1-基)-2-溴丙基-2-烯-1-酮的制备(化合物(S)-5-3)Example 38. (S)-1-(3-(4-Amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidine Preparation of 1-yl)-2-bromopropyl-2-en-1-one (Compound (S)-5-3)
Figure PCTCN2016081669-appb-000050
Figure PCTCN2016081669-appb-000050
合成步骤参考实施例7.用类似于化合物5-1的合成方法，将2-氟丙烯酸替换为2-溴丙烯酸制备化合物5-3(225mg，产率33％，LC-ESI-MS：[M+H]＝519.2)。Synthesis procedure Reference Example 7. Compound 5-3 (225 mg, yield 33%, LC-ESI-MS: [M. +H]=519.2).
1H-NMR(δ,CDCl3):8.35(s,1H,ArH),7.63(d,2H,J＝8.0Hz,ArH),7.39(m,2H,ArH),7.14(m,3H,ArH),7.10(d,2H,J＝8.0Hz,ArH),6.04(s,1H,-NH2),5.78(m,3H,-C＝CH2,-NH2),4.92(m,1H,piperidine),4.66(m,0.5H,piperidine),4.52(m,0.5H,piperidine),4.13(m,0.5H,piperidine),3.97(m,0.5H,piperidine),3.78(m,0.5H,piperidine),3.21(m,0.5H,piperidine),3.11(m,0.5H,piperidine),2.92(m,0.5H,piperidine),2.26(m,2H,piperidine),1.72(m,2H,piperidine). 1 H-NMR (δ, CDCl 3 ): 8.35 (s, 1H, ArH), 7.63 (d, 2H, J = 8.0 Hz, ArH), 7.39 (m, 2H, ArH), 7.14 (m, 3H, ArH) ), 7.10 (d, 2H, J = 8.0 Hz, ArH), 6.04 (s, 1H, -NH 2 ), 5.78 (m, 3H, -C=CH 2 , -NH 2 ), 4.92 (m, 1H, Piperidine), 4.66 (m, 0.5H, piperidine), 4.52 (m, 0.5H, piperididine), 4.13 (m, 0.5H, piperididine), 3.97 (m, 0.5H, piperididine), 3.78 (m, 0.5H, Piperidine), 3.21 (m, 0.5H, piperididine), 3.11 (m, 0.5H, piperididine), 2.92 (m, 0.5H, piperididine), 2.26 (m, 2H, piperididine), 1.72 (m, 2H, piperididine) .
实施例39.(S)-1-(3-(4-氨基-3-(6-苯氧基吡啶-3-基)-1H-吡唑并[3,4-d]嘧啶-1-基)哌啶-1-基)-2-溴丙基-2-烯-1-酮的制备(化合物(S)-5-19) Example 39. (S)-1-(3-(4-Amino-3-(6-phenoxypyridin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl Preparation of piperidin-1-yl)-2-bromopropyl-2-en-1-one (compound (S)-5-19)
Figure PCTCN2016081669-appb-000051
Figure PCTCN2016081669-appb-000051
合成步骤参考实施例7.用类似于化合物5-1的合成方法，将化合物4a替换为(S)-4g，将2-氟丙烯酸替换为2-溴丙烯酸，制备化合物(S)-5-19(242mg，产率36％，LC-ESI-MS：[M+H]＝520.2)。Synthesis procedure Reference Example 7. Compound (S)-5-19 was prepared by a synthesis method similar to Compound 5-1, replacing Compound 4a with (S)-4g and 2-fluoroacrylic acid with 2-bromoacrylic acid. (242 mg, yield 36%, LC-ESI-MS: [M+H]=520.2).
1H-NMR(δ,CDCl3):8.52(d,1H,J＝2.0Hz,ArH),8.36(s,1H,ArH),8.03(dd,1H,J1＝8.0Hz,J2＝2.4Hz,ArH),7.421(td,2H,J1＝7.6Hz,J2＝2.0Hz,ArH),7.21(t,1H,J＝8.0Hz,ArH),7.19(dd,2H,J1＝8.4Hz,J2＝2.0Hz,ArH),7.09(d,1H,J＝8.4Hz,ArH),6.06(s,2H,-NH2),5.18(d,0.5H,J＝4.0Hz,-C＝CH2),5.10(d,0.5H,J＝4.0Hz,-C＝CH2),5.05(d,1H,-C＝CH2),4.91(m,1H,piperidine),4.63(m,0.5H,piperidine),4.45(m,0.5H,piperidine),4.11(m,0.5H,piperidine),3.75(m,0.5H,piperidine),3.44(m,0.5H,piperidine),3.17(m,0.5H,piperidine),2.85(m,0.5H,piperidine),2.79(m,0.5H,piperidine),2.32(m,2H,piperidine),1.72(m,2H,piperidine). 1 H-NMR (δ, CDCl 3 ): 8.52 (d, 1H, J = 2.0 Hz, ArH), 8.36 (s, 1H, ArH), 8.03 (dd, 1H, J 1 = 8.0 Hz, J 2 = 2.4 Hz, ArH), 7.421 (td, 2H, J 1 = 7.6 Hz, J 2 = 2.0 Hz, ArH), 7.21 (t, 1H, J = 8.0 Hz, ArH), 7.19 (dd, 2H, J 1 = 8.4 Hz, J 2 = 2.0 Hz, ArH), 7.09 (d, 1H, J = 8.4 Hz, ArH), 6.06 (s, 2H, -NH 2 ), 5.18 (d, 0.5H, J = 4.0 Hz, -C =CH 2 ), 5.10 (d, 0.5H, J = 4.0 Hz, -C=CH 2 ), 5.05 (d, 1H, -C=CH 2 ), 4.91 (m, 1H, piperididine), 4.63 (m, 0.5H,piperidine), 4.45 (m, 0.5H, piperididine), 4.11 (m, 0.5H, piperididine), 3.75 (m, 0.5H, piperididine), 3.44 (m, 0.5H, piperididine), 3.17 (m, 0.5H,piperidine), 2.85 (m, 0.5H, piperididine), 2.79 (m, 0.5H, piperididine), 2.32 (m, 2H, piperididine), 1.72 (m, 2H, piperididine).
实施例40.(S)-1-(3-(4-氨基-3-(6-苯氧基吡啶-3-基)-1H-吡唑并[3,4-d]嘧啶-1-基)哌啶-1-基)-2-氯丙基-2-烯-1-酮的制备(化合物(S)-5-20)Example 40. (S)-1-(3-(4-Amino-3-(6-phenoxypyridin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl Preparation of piperidin-1-yl)-2-chloropropyl-2-en-1-one (compound (S)-5-20)
Figure PCTCN2016081669-appb-000052
Figure PCTCN2016081669-appb-000052
合成步骤参考实施例7.用类似于化合物5-1的合成方法，将化合物4a替换为(S)-4g，将2-氟丙烯酸替换为2-氯丙烯酸，制备化合物(S)-5-20(300mg，产率49％，LC-ESI-MS：[M+H]＝476.1)。Synthesis procedure Reference Example 7. Compound (S)-5-20 was prepared by a synthesis method similar to Compound 5-1, replacing Compound 4a with (S)-4g and 2-fluoroacrylic acid with 2-chloroacrylic acid. (300 mg, yield 49%, LC-ESI-MS: [M+H] = 476.1).
1H-NMR(δ,CDCl3):8.52(d,1H,J＝2.0Hz,ArH),8.37(s,1H,ArH),8.05(dd,1H,J1＝8.0Hz,J2＝2.0Hz,ArH),7.45(td,2H,J1＝8.0Hz,J2＝2.0Hz,ArH),7.26(t,1H,J＝7.6Hz,ArH),7.22(dd,2H,J1＝8.4Hz,J2＝2.0Hz,ArH),7.12(d,1H,J＝8.4Hz,ArH),5.99(s,2H,-NH2),5.23(d,0.5H,J＝4.0Hz,-C＝CH2),5.11(d,0.5H,J＝4.0Hz,-C＝CH2),5.06(d,1H,-C＝CH2),4.92(m,1H,piperidine),4.63(m,0.5H,piperidine),4.43(m,0.5H,piperidine),4.12(m,0.5H,piperidine),3.74(m,0.5H,piperidine),3.42(m,0.5H,piperidine),3.18(m,0.5H,piperidine),2.81(m,0.5H,piperidine),2.77(m,0.5H,piperidine),2.34(m,2H,piperidine),1.73(m,2H,piperidine). 1 H-NMR (δ, CDCl 3 ): 8.52 (d, 1H, J = 2.0 Hz, ArH), 8.37 (s, 1H, ArH), 8.05 (dd, 1H, J 1 = 8.0 Hz, J 2 = 2.0 Hz, ArH), 7.45 (td, 2H, J 1 = 8.0 Hz, J 2 = 2.0 Hz, ArH), 7.26 (t, 1H, J = 7.6 Hz, ArH), 7.22 (dd, 2H, J 1 = 8.4 Hz, J 2 = 2.0 Hz, ArH), 7.12 (d, 1H, J = 8.4 Hz, ArH), 5.99 (s, 2H, -NH 2 ), 5.23 (d, 0.5H, J = 4.0 Hz, -C =CH 2 ), 5.11 (d, 0.5H, J = 4.0 Hz, -C=CH 2 ), 5.06 (d, 1H, -C=CH 2 ), 4.92 (m, 1H, piperididine), 4.63 (m, 0.5H,piperidine), 4.43 (m, 0.5H, piperididine), 4.12 (m, 0.5H, piperididine), 3.74 (m, 0.5H, piperididine), 3.42 (m, 0.5H, piperididine), 3.18 (m, 0.5H,piperidine), 2.81 (m, 0.5H, piperididine), 2.77 (m, 0.5H, piperididine), 2.34 (m, 2H, piperididine), 1.73 (m, 2H, piperididine).
实施例41.体外Btk激酶抑制活性和体外抗肿瘤活性测试Example 41. In vitro Btk kinase inhibitory activity and in vitro antitumor activity assay
本发明化合物的体外Btk激酶抑制活性测定方法： In vitro Btk kinase inhibitory activity assay for compounds of the invention:
将药物溶于DMSO中制成10mM的储备液，并稀释至50x的测试浓度药液备用，测试浓度以3被梯度稀释，分别为25nM，8.33nM，2.78nM，0.93nM，0.31nM，0.10nM。在96孔板里加入10μL 50x药物备用液，再加入90μL 1x激酶缓冲液，在振荡器上震荡10分钟。从96孔板各孔取5μL转移至384孔板，384孔板里设2复孔。激酶反应：准备2.5x激酶缓冲液：将酶加入1x激酶基础缓冲液中。准备2.5x短肽溶液：将FAM标记的短肽和ATP加入1x激酶基础缓冲液中。在加有5μL药液384孔板中，加入10μL 2.5x激酶缓冲液，室温孵育10分钟。在384孔板中加入10μL 2.5x短肽溶液，28℃孵育1小时。加入25μL终止液停止反应。读数，并计算化合物对酶的抑制率，拟合计算出BTK激酶的IC50。The drug was dissolved in DMSO to make a 10 mM stock solution, and diluted to a 50x test concentration. The test concentration was diluted by 3 to 25nM, 8.33nM, 2.78nM, 0.93nM, 0.31nM, 0.10nM. . 10 μL of 50x drug stock solution was added to a 96-well plate, and 90 μL of 1x kinase buffer was added and shaken on the shaker for 10 minutes. Transfer 5 μL from each well of a 96-well plate to a 384-well plate, and set 2 duplicate wells in a 384-well plate. Kinase Reaction: Prepare 2.5x Kinase Buffer: Add the enzyme to the 1x Kinase Buffer. Prepare a 2.5x short peptide solution: Add the FAM-labeled short peptide and ATP to the 1x Kinase Buffer. In a 384-well plate supplemented with 5 μL of drug solution, 10 μL of 2.5x kinase buffer was added and incubated for 10 minutes at room temperature. 10 μL of a 2.5× short peptide solution was added to a 384-well plate and incubated at 28 ° C for 1 hour. The reaction was stopped by adding 25 μL of stop solution. Readings, and calculate the enzyme inhibition ratio of the compound, the calculated IC 50 fit BTK kinases.
选用不同的实体瘤和白血病细胞株对所合成的化合物进行了体外抗肿瘤活性的测定:The antitumor activity of the synthesized compounds was determined by using different solid tumors and leukemia cell lines:
细胞株：人肺癌细胞(A549、PC9)、人乳腺癌细胞(MCF7)、人胃癌细胞(SGC7901)、急性前髓细胞性白血病细胞(HL60)、人肺癌细胞吉非替尼耐药细胞(PC9-IR)。Cell lines: human lung cancer cells (A549, PC9), human breast cancer cells (MCF7), human gastric cancer cells (SGC7901), acute promyelocytic leukemia cells (HL60), human lung cancer cells gefitinib-resistant cells (PC9) -IR).
培养基：A549:RPMI 1640+胎牛血清Medium: A549: RPMI 1640 + fetal bovine serum
        MCF7：DMEM+胎牛血清MCF7: DMEM + fetal bovine serum
        SGC7901：RPMI 1640+新生牛血清SGC7901: RPMI 1640+ newborn calf serum
        HL60：RPMI 1640+胎牛血清HL60: RPMI 1640+ fetal calf serum
        PC9：DMEM+胎牛血清PC9: DMEM + fetal bovine serum
        PC9-IR：DMEM+胎牛血清PC9-IR: DMEM + fetal bovine serum
药物配制方法：将药物溶于DMSO中制成10mM的储备液，并按一定比例稀释得到5个不同浓度(测试浓度100x)。Drug preparation method: The drug was dissolved in DMSO to make a 10 mM stock solution, and diluted to a certain ratio to obtain 5 different concentrations (test concentration 100x).
肿瘤细胞体外培养：Tumor cell culture in vitro:
将所选取的六株肿瘤细胞A549、MCF7、SGC7901、HL60、PC9、PC9-IR，于37℃、5％CO2细胞培养箱中孵育，待细胞密度长到70～90％时传代(贴壁细胞用Duck’s EDTA消化后传代)，用于以后实验所需。The selected six tumor cells A549, MCF7, SGC7901, HL60, PC9, PC9-IR were incubated in a 37 ° C, 5% CO 2 cell incubator, and were passaged when the cell density was 70-90%. Cells were passaged with Duck's EDTA and passaged for later experiments.
肿瘤细胞A549、MCF7、SGC7901、HL60、PC9、PC9-IR，在96孔板上种入4000个/200μL/孔，于37℃、5％CO2细胞培养箱中孵育过夜。每孔加入化合物2μL，终浓度为50μM，10μM，2μM，0.4μM，0.08μM共同于37℃、5％CO2细胞培养箱中孵育72小时，以DMSO(2％)为对照组。72小时后，加入20μL CCK-8溶液，置于37℃、5％CO2细胞培养箱中4小时。用加了相应量细胞培养液和CCK-8溶液但没有加入细胞的孔作为空白对照。用酶标仪在450nm测定吸光度(OD值)，所得数据用于计算IC50。Tumor cells A549, MCF7, SGC7901, HL60, PC9, PC9-IR were seeded in a 96-well plate at 4000 cells/200 μL/well and incubated overnight at 37 ° C in a 5% CO 2 cell incubator. 2 μL of compound was added to each well to a final concentration of 50 μM, 10 μM, 2 μM, 0.4 μM, and 0.08 μM for 72 hours at 37 ° C in a 5% CO 2 cell incubator with DMSO (2%) as a control. After 72 hours, 20 μL of CCK-8 solution was added and placed in a 37 ° C, 5% CO 2 cell incubator for 4 hours. Wells containing the corresponding amount of cell culture medium and CCK-8 solution but no cells were added as a blank control. The absorbance (OD value) was measured at 450 nm using a microplate reader, and the obtained data was used to calculate the IC 50 .
细胞抑制率的计算公式为：细胞抑制率％＝[(对照组OD值-空白组OD值)-(用药组OD值-空白组OD值)]/(对照细胞OD值-空白组OD值)×100％，用CalcuSyn软件计算求得半数抑制浓度(IC50)。The formula for calculating the cell inhibition rate is: cell inhibition rate % = [(control group OD value - blank group OD value) - (medical group OD value - blank group OD value)] / (control cell OD value - blank group OD value) ×100%, the half-inhibitory concentration (IC 50 ) was calculated by CalcuSyn software.
表1 部分化合物对BTK激酶抑制活性和体外肿瘤细胞增殖抑制活性Table 1 Some compounds inhibit BTK kinase activity and inhibit tumor cell proliferation in vitro
Figure PCTCN2016081669-appb-000053
Figure PCTCN2016081669-appb-000053
Figure PCTCN2016081669-appb-000054
Figure PCTCN2016081669-appb-000054
N.T.为未测试。N.T. is not tested.
从表1中数据可以看出，所有的化合物对BTK都具有明显的抑制活性，活性均小于10nM，与阳性对照伊布替尼活性相当，说明在丙烯酰胺的α位引入卤素对其活性基本没有影响，仍然表现出强效的BTK激酶抑制活性。但是，在细胞水平上，所测试的化合物5-1～5-6和5-13～5-24均表现出比阳性对照更为强效的肿瘤细胞增殖抑制活性，不仅仅对实体瘤细胞活性更强(A549、SGC-7901、MCF-7、PC-9)，且对血液瘤细胞(HL-60)同样如此。譬如，化合物5-2在SGC-7901细胞上活性高了5倍。同时，所有化合物在PC9-IR(吉非替尼耐药的PC-9细胞株)中同样显示出强效的肿瘤细胞增殖抑制活性。因此，本发明所涉及的可用作BTK抑制剂具有广阔的抗肿瘤应用前景。It can be seen from the data in Table 1 that all the compounds have significant inhibitory activity against BTK, and the activities are less than 10 nM, which is equivalent to the activity of the positive control, ibutinab, indicating that the introduction of halogen in the alpha position of acrylamide has substantially no activity. Effects, still showing potent BTK kinase inhibitory activity. However, at the cellular level, the tested compounds 5-1 to 5-6 and 5-13 to 5-24 all showed more potent tumor cell proliferation inhibitory activity than the positive control, not only for solid tumor cells. Stronger (A549, SGC-7901, MCF-7, PC-9), and the same for hematoma cells (HL-60). For example, Compound 5-2 is 5 times more active on SGC-7901 cells. At the same time, all of the compounds also showed potent tumor cell proliferation inhibitory activity in PC9-IR (gefitinib-resistant PC-9 cell strain). Therefore, the BTK inhibitors involved in the present invention have broad anti-tumor application prospects.
实施例42.溶解度试验Example 42. Solubility test
称取研成细粉的受试化合物100mg，分别于25℃±2℃在一定量的溶剂(1ml，3ml，10ml，100ml)中，每隔5分钟强力振摇30秒钟；观察30分钟内的溶解情况，如无可见溶质，视为完全溶解。100 mg of the test compound which was ground into a fine powder was weighed and shaken vigorously for 30 seconds every 5 minutes at 25 ° C ± 2 ° C in a certain amount of solvent (1 ml, 3 ml, 10 ml, 100 ml); The dissolution, if no visible solutes, is considered complete dissolution.
实验结果显示，化合物5-3在100ml溶剂时仍有部分未溶解，参照中国药典的定义，属于极微溶解。而化合物5-19在100ml溶剂时完全溶解，在10ml时有小部分未溶解，属于微溶，比化合物5-3明显提高。说明芳香环氮原子的引入明显提高了化合物的溶解度。The experimental results show that the compound 5-3 is still partially dissolved in 100 ml of the solvent, and is extremely slightly soluble as defined by the Chinese Pharmacopoeia. Compound 5-19 was completely dissolved in 100 ml of solvent, and a small portion was undissolved at 10 ml, which was slightly soluble and significantly higher than compound 5-3. It is indicated that the introduction of the aromatic ring nitrogen atom significantly increases the solubility of the compound.
实施例43.体外双位点不可逆抑制剂性质的表征：Example 43. Characterization of in vitro two-site irreversible inhibitor properties:
首先，采用化合物5-3预处理重组Btk后，用不含抑制剂的培养基重复洗涤，其活性不会恢复(参见例如J.B.Smaill等，J.Med.Chem.1999,42，1803)。其次，通过质谱，观察到了主要质谱峰对应化合物5-3和Btk之间的共价复合物的分子量之比为1:1(化合物4:518,道尔顿，重组Btk激酶结合域：33487；复合物预测值为34005，实测值为34005)。First, after pretreatment of recombinant Btk with compound 5-3, repeated washing with medium without inhibitor does not restore its activity (see, for example, J. B. Smaill et al., J. Med. Chem. 1999, 42, 1803). Secondly, by mass spectrometry, it was observed that the ratio of the molecular weight of the main mass spectrum peak corresponding to the covalent complex between compound 5-3 and Btk was 1:1 (compound 4:518, Dalton, recombinant Btk kinase binding domain: 33487; The predicted value of the composite was 34,005 and the measured value was 34,005).
其次，采用化合物5-3与半胱氨酸共孵浴3小时后，经HPLC-MS检测，发现大部分化合物5-3转变为被两个半胱氨酸加成的产物，经检测体系中存在目标产物(5-3的反应体系，LC-ESI-MS：[M+H]681)。因此，所测试的化合物具有被双分子含巯基化合物加成的能力，这 也阐明了此类化合物虽然在体外Btk激酶抑制活性与阳性药伊布替尼相当，但是在细胞层面上能明显优于伊布替尼。Secondly, after compound 5-3 was incubated with cysteine for 3 hours, it was found by HPLC-MS that most of the compound 5-3 was converted into a product added by two cysteines. The target product (5-3 reaction system, LC-ESI-MS: [M+H] 681) was obtained. Therefore, the tested compound has the ability to be added by a bimolecular thiol-containing compound, which It is also clarified that although the in vitro Btk kinase inhibitory activity is comparable to the positive drug ibutinib, it is significantly superior to ibupotinib at the cellular level.
实施例44.口服药代动力学实验Example 44. Oral pharmacokinetic experiment
以伊布替尼为参比，分别考察了化合物5-2、5-3和5-19在大鼠体内的药代动力学性质，具体方法如下：以SD大鼠为实验动物，灌胃给药20mg/kg，尾静脉静注给药5mg/kg。灌胃给药的尾静脉取血时间点为0.17,0.33,0.5,1,1.5,2,4,6,8,12,24小时；静脉给药取血时间点为0.05,0.1,0.17,0.5,1,2,4,6,8,12,24小时。取全血0.3ml，离心后取血浆0.1ml采用LC-MS进行分析。结果表明，化合物5-2、5-3和5-19的口服生物利用度分别为25％、17％和19％，而伊布替尼的生物利用度为12％，因此化合物5-2、5-3和5-19在体内的稳定性较伊布替尼有了明显的显著提高。其原因可能在于丙烯酰胺的α位引入卤素引入后，在不影响药物吸收的前提下，使得双键的代谢速率受到一定程度的抑制，进而提高了血药浓度而提高生物利用度。Taking ibbutinib as a reference, the pharmacokinetic properties of compounds 5-2, 5-3 and 5-19 in rats were investigated. The specific methods were as follows: SD rats were used as experimental animals, and given by gavage The drug was administered at a dose of 20 mg/kg, and 5 mg/kg was administered intravenously to the tail vein. The time of blood collection from the tail vein of the intragastric administration was 0.17, 0.33, 0.5, 1, 1.5, 2, 4, 6, 8, 12, 24 hours; the time of blood collection by intravenous administration was 0.05, 0.1, 0.17, 0.5. 1,2,4,6,8,12,24 hours. 0.3 ml of whole blood was taken, and 0.1 ml of plasma was taken after centrifugation and analyzed by LC-MS. The results showed that the oral bioavailabilities of compounds 5-2, 5-3, and 5-19 were 25%, 17%, and 19%, respectively, while the bioavailability of Ibubinib was 12%, so compound 5-2, The stability of 5-3 and 5-19 in vivo was significantly improved compared to ibufenib. The reason may be that after the introduction of halogen into the α-position of acrylamide, the metabolic rate of the double bond is inhibited to a certain extent without affecting the absorption of the drug, thereby increasing the plasma concentration and improving the bioavailability.
实施例45：使用化合物5-2和5-19治疗类分湿性关节炎Example 45: Treatment of differential arthritis with compounds 5-2 and 5-19
在Balb/c小鼠中，通过给予抗胶原蛋白抗体和脂多糖诱导关节炎(Nandakumar等，Am.J.Pathol.2003,163:1827-1837)。具体方法如下：在第0天，在雌性Balb/c小鼠静脉内注射100mg/kg抗II型胶原蛋白的Chemico mAb合剂，在第1天，腹膜内注射1.25mg/kg脂多糖。在第2天至12天，按10mg/kg的化合物5-2和5-19，每天口服给药1次。结果表明，化合物5-2和5-19均具有明显的体内抗类分湿性关节炎的作用，具体来讲，模型组中出现的炎性细胞浸润、滑膜增生、炎性肉芽组织形成，坏死组织出现等现象在化合物5-2和5-19治疗后得到明显改善。 In Balb/c mice, arthritis is induced by administration of an anti-collagen antibody and lipopolysaccharide (Nandakumar et al, Am. J. Pathol. 2003, 163: 1827-1837). The specific method was as follows: On day 0, female Balb/c mice were intravenously injected with 100 mg/kg of anti-type II collagen Chemico mAb mixture, and on day 1, 1.25 mg/kg lipopolysaccharide was intraperitoneally injected. On days 2 to 12, 10 mg/kg of Compounds 5-2 and 5-19 were administered orally once a day. The results showed that both compounds 5-2 and 5-19 had significant anti-hypertensive arthritis effects in vivo. Specifically, inflammatory cell infiltration, synovial hyperplasia, inflammatory granulation tissue formation, necrosis occurred in the model group. The appearance of tissue and the like were significantly improved after treatment with compounds 5-2 and 5-19.

Claims (11)

  1. 一种布鲁顿酪氨酸激酶抑制剂，其特征在于，其具有通式Ⅰ的结构：A Bruton tyrosine kinase inhibitor characterized by having the structure of Formula I:
    Figure PCTCN2016081669-appb-100001
    Figure PCTCN2016081669-appb-100001
    或其光学异构体或其药学上可接受的盐或溶剂合物，其中：Ra,Rb,Rc独立选自H、卤素、-CF3、-CN、-NO2、OH、NH2、-L-C1-C6的烷基、-L-C1-C6的烯基、-L-取代或非取代的杂芳基、或-L-取代或非取代的芳基，其中L是键、O、S、-S(＝O)、-S(＝O)2、NH、C(O)、CH2、-NHC(O)O、-NHC(O)或-C(O)NH，X选自氟、氯或溴，Y1,Y2独立选自C、N。Or an optical isomer thereof, or a pharmaceutically acceptable salt or solvate thereof, wherein: Ra, Rb, Rc are independently selected from the group consisting of H, halogen, -CF 3 , -CN, -NO 2 , OH, NH 2 , - LC alkyl, -LC 1 -C 6 alkenyl group of the 1 -C 6, -L- aryl substituted or unsubstituted heteroaryl group, or -L-substituted or unsubstituted aryl group, wherein L is a bond, O, S, -S(=O), -S(=O) 2 , NH, C(O), CH 2 , -NHC(O)O, -NHC(O) or -C(O)NH, X is selected from Fluorine, chlorine or bromine, Y 1 , Y 2 are independently selected from C and N.
  2. 一种布鲁顿酪氨酸激酶抑制剂，其特征在于，其具有通式II的结构：A Bruton tyrosine kinase inhibitor characterized by having the structure of Formula II:
    Figure PCTCN2016081669-appb-100002
    Figure PCTCN2016081669-appb-100002
    或其光学异构体或其药学上可接受的盐或溶剂合物，其中：Rd1、Rd2、Rd3、Rd4、Rd5独立地是H、卤素、-CF3、-CN、-NO2、-OH、C1-C3的烷氧基或-NH2，X选自氟、氯或溴。Or an optical isomer or a pharmaceutically acceptable salt or solvate thereof, wherein: Rd 1, Rd 2, Rd 3, Rd 4, Rd 5 is independently H, halo, -CF 3, -CN, - NO 2 , -OH, C 1 -C 3 alkoxy or -NH 2 , X is selected from fluorine, chlorine or bromine.
  3. 一种布鲁顿酪氨酸激酶抑制剂，其特征在于，其具有通式III的结构： A Bruton tyrosine kinase inhibitor characterized by having the structure of Formula III:
    Figure PCTCN2016081669-appb-100003
    Figure PCTCN2016081669-appb-100003
    或其光学异构体或其药学上可接受的盐或溶剂合物，其中：X选自氟、氯或溴。Or an optical isomer thereof, or a pharmaceutically acceptable salt or solvate thereof, wherein: X is selected from the group consisting of fluorine, chlorine or bromine.
  4. 一种布鲁顿酪氨酸激酶抑制剂，其特征在于，其具有通式IV的结构：A Bruton tyrosine kinase inhibitor characterized by having the structure of Formula IV:
    Figure PCTCN2016081669-appb-100004
    Figure PCTCN2016081669-appb-100004
    或其光学异构体或其药学上可接受的盐或溶剂合物，其中：Rd1、Rd2、Rd3、Rd4、Rd5独立地是H、卤素、-CF3、-CN、-NO2、-OH、C1-C3的烷氧基、或-NH2，X选自氟、氯或溴，Y1,Y2独立选自C、N，Y1,Y2至少有一个选自N。Or an optical isomer or a pharmaceutically acceptable salt or solvate thereof, wherein: Rd 1, Rd 2, Rd 3, Rd 4, Rd 5 is independently H, halo, -CF 3, -CN, - NO 2 , -OH, C 1 -C 3 alkoxy group, or -NH 2 , X is selected from fluorine, chlorine or bromine, Y 1 , Y 2 are independently selected from C, N, Y 1 , and Y 2 has at least one From N.
  5. 一种布鲁顿酪氨酸激酶抑制剂，其特征在于，其具有通式V的结构： A Bruton tyrosine kinase inhibitor characterized by having the structure of Formula V:
    Figure PCTCN2016081669-appb-100005
    Figure PCTCN2016081669-appb-100005
    或其光学异构体或其药学上可接受的盐或溶剂合物，其中：X选自氟、氯或溴,Y1,Y2独立选自C、N，Y1,Y2至少有一个选自N。Or an optical isomer thereof or a pharmaceutically acceptable salt or solvate thereof, wherein: X is selected from fluorine, chlorine or bromine, and Y 1 , Y 2 are independently selected from C, N, Y 1 and Y 2 are at least one From N.
  6. 一种药物组合物，所述药物组合物包含至少一种活性组分以及一种或多种药学上可接受的载体或赋形剂，所述活性组分是如权利要求1至5中任意一项所述的布鲁顿酪氨酸激酶抑制剂化合物、所述化合物的光学异构体、所述化合物或其光学异构体在药学上可接受的盐、所述化合物或其光学异构体的溶剂合物中的任意一种或任意多种。A pharmaceutical composition comprising at least one active ingredient together with one or more pharmaceutically acceptable carriers or excipients, said active ingredient being any one of claims 1 to 5 Or a pharmaceutically acceptable salt, the compound or an optical isomer thereof Any one or any of a plurality of solvates.
  7. 如权利要求1至5中任意一项所述的布鲁顿酪氨酸激酶抑制剂化合物及其光学异构体或其药学上可接受的盐或溶剂合物在制备治疗除干眼症外的从布鲁顿酪氨酸激酶活性的抑制中获益的疾病、障碍或病症药物中的应用。The Bruton tyrosine kinase inhibitor compound according to any one of claims 1 to 5, and an optical isomer thereof, or a pharmaceutically acceptable salt or solvate thereof, for the preparation of a treatment other than dry eye Use in medicines for diseases, disorders or conditions that benefit from inhibition of Bruton's tyrosine kinase activity.
  8. 如权利要求1至5中任意一项所述的布鲁顿酪氨酸激酶抑制剂化合物及其光学异构体或其药学上可接受的盐或溶剂合物在制备单独或和其他药物联合使用治疗细胞增生疾病药物中的应用。The Bruton tyrosine kinase inhibitor compound according to any one of claims 1 to 5, and an optical isomer thereof, or a pharmaceutically acceptable salt or solvate thereof, for use alone or in combination with other drugs The application of drugs for treating cell proliferative diseases.
  9. 如权利要求1至5中任意一项所述的布鲁顿酪氨酸激酶抑制剂化合物及其光学异构体或其药学上可接受的盐或溶剂合物在制备单独或和其他药物联合使用治疗癌症药物中的应用。The Bruton tyrosine kinase inhibitor compound according to any one of claims 1 to 5, and an optical isomer thereof, or a pharmaceutically acceptable salt or solvate thereof, for use alone or in combination with other drugs Application in the treatment of cancer drugs.
  10. 如权利要求1至5中任意一项所述的布鲁顿酪氨酸激酶抑制剂化合物及其光学异构体或其药学上可接受的盐或溶剂合物在制备单独或和其他药物联合使用治疗除干眼症外的自身免疫性疾病药物中的应用。The Bruton tyrosine kinase inhibitor compound according to any one of claims 1 to 5, and an optical isomer thereof, or a pharmaceutically acceptable salt or solvate thereof, for use alone or in combination with other drugs The application in the treatment of autoimmune diseases other than dry eye.
  11. 如权利要求1至5中任意一项所述的布鲁顿酪氨酸激酶抑制剂化合物及其光学异构体或其药学上可接受的盐或溶剂合物在制备单独或和其他药物联合使用***药物中的应用。 The Bruton tyrosine kinase inhibitor compound according to any one of claims 1 to 5, and an optical isomer thereof, or a pharmaceutically acceptable salt or solvate thereof, for use alone or in combination with other drugs Application in the treatment of lupus erythematosus drugs.
PCT/CN2016/081669 2015-05-12 2016-05-11 Dual-site irreversible bruton's tyrosine kinase inhibitor, composition and application therefof WO2016180334A1 (en)

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