WO2016045598A1 - 4-substituted pyrrolo[2,3-d]pyrimidine compound and use thereof - Google Patents

4-substituted pyrrolo[2,3-d]pyrimidine compound and use thereof Download PDF

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WO2016045598A1
WO2016045598A1 PCT/CN2015/090497 CN2015090497W WO2016045598A1 WO 2016045598 A1 WO2016045598 A1 WO 2016045598A1 CN 2015090497 W CN2015090497 W CN 2015090497W WO 2016045598 A1 WO2016045598 A1 WO 2016045598A1
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compound
pharmaceutically acceptable
group
tautomer
pyrrole
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PCT/CN2015/090497
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French (fr)
Chinese (zh)
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秦引林
苏梅
金秋
陈涛
伍贤志
蒋建华
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江苏柯菲平医药股份有限公司
南京柯菲平盛辉制药有限公司
南京柯菲平制药有限公司
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Publication of WO2016045598A1 publication Critical patent/WO2016045598A1/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
    • 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
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/4353—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems
    • A61K31/437—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems the heterocyclic ring system containing a five-membered ring having nitrogen as a ring hetero atom, e.g. indolizine, beta-carboline
    • 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
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/4353—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems
    • A61K31/4375—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems the heterocyclic ring system containing a six-membered ring having nitrogen as a ring heteroatom, e.g. quinolizines, naphthyridines, berberine, vincamine
    • 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 present invention relates to a 4-substituted pyrrolo[2,3-d]pyrimidine compound and its use in the preparation of a medicament for the treatment of diseases of the immune system, treatment of rheumatoid diseases such as rheumatoid, tumors and the like.
  • Protein kinases also known as protein phosphakinase, are a class of enzymes that catalyze the phosphorylation of proteins. It can transfer the ⁇ -phosphate on adenosine triphosphate (ATP) to the amino acid residues of protein molecules, thereby changing the conformation and activity of proteins and enzymes. Phosphorylation of proteins is an important part of many signaling pathways, and most important life processes in cells are inseparable from protein phosphorylation. These enzymes are key factors in regulating cellular signaling, including cell proliferation and cell differentiation.
  • ATP adenosine triphosphate
  • Protein kinase signaling plays a major role in transduction: one is to regulate protein activity through phosphorylation. Phosphorylation and dephosphorylation are common mechanisms for reversible activation of most signaling pathway components. Some proteins are phosphorylated. It has activity, some is active after dephosphorylation; the other is to gradually amplify the signal through the stepwise phosphorylation of the protein, causing cell reaction.
  • Janus-activated kinase Singal transducers and activators of transcriprion is a newly discovered intracellular signaling pathway closely related to cytokines, which is involved in cell proliferation and differentiation. Many important biological processes such as apoptosis and immune regulation.
  • Janus kinase is a non-receptor tyrosine protein kinase.
  • JAK1, JAK2, TYK2 and JAK3 There are 4 family members, namely JAK1, JAK2, TYK2 and JAK3.
  • JAK is a very important drug target
  • JAK inhibitors While JAK is a very important drug target, JAK inhibitors have been proven to be useful in the treatment of blood system diseases, tumors, rheumatoid arthritis and psoriasis. Because JAK inhibitors have significant medical uses and can be used in a variety of related disease drugs, the research and discovery of such compounds is extremely beneficial.
  • JAK inhibitor of a related disease Its mother core structure is shown in the following chemical formula.
  • As an effective JAK inhibitor it can be used as a drug substance for the preparation of a medicament for treating diseases such as rheumatoid arthritis, skin diseases, cancer, and myeloproliferative diseases.
  • JAK inhibitors which is marketed as a typical JAK inhibitor, is used for rheumatoid arthritis (rheumatoid). Arthritis). The first drug to treat.
  • JAK inhibitors involves a wide range of applications, and it is an effort in the field to seek new, more active, and more highly compounded compounds.
  • a first technical object of the present invention is to provide a novel JAK inhibitor compound; a second technical object of the present invention is to provide a JAK inhibitor compound of the present invention in the preparation of a medicament for treating a disease associated with a JAK inhibitor application.
  • R 1 is selected from the group consisting of hydrogen, halogen, and alkyl.
  • R 2 is selected from: Wherein R 4 , R 5 , R 6 , and R 7 are each independently selected from the group consisting of hydrogen, halogen, alkyl, nitro, amino, and hydroxy; and X is selected from O, N or S. Further, in R 2 , R 4 , R 5 , R 6 and R 7 are each independently selected from the group consisting of hydrogen, halogen and alkyl.
  • R 3 is selected from: Wherein R 8 is selected from the group consisting of alkyl, cycloalkyl and heterocycloalkyl.
  • R 3 is selected from
  • a pharmaceutical composition comprising a therapeutically effective amount of a compound of any one of formula (I), a stereoisomer, tautomer or pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier , adjuvant or solvent.
  • a compound represented by the formula (I), a stereoisomer, a tautomer, a prodrug or a pharmaceutically acceptable salt thereof is used as a medicament.
  • a compound of the formula (I), a stereoisomer, a tautomer, a prodrug or a pharmaceutically acceptable salt thereof which is used as a medicament for treating an immune disease; and a treatment selected from rheumatoid sex Drugs for arthritis, asthma, systemic lupus erythematosus, psoriasis, IBD, and transplant rejection of immune diseases.
  • JAK1 kinase activity for inhibiting a responsive disease, particularly an immune disease, more particularly selected from the group consisting of rheumatoid arthritis, asthma, systemic lupus erythematosus, psoriasis, IBD, and transplant rejection Immune disease.
  • a responsive disease particularly an immune disease, more particularly selected from the group consisting of rheumatoid arthritis, asthma, systemic lupus erythematosus, psoriasis, IBD, and transplant rejection Immune disease.
  • Alkyl means a saturated aliphatic hydrocarbon group. A straight or branched chain group of 1 to 20 carbon atoms is included. Preference is given to medium-sized alkyl groups having 1 to 6 carbon atoms, such as methyl, ethyl, propyl, 2-propyl, n-butyl, isobutyl, tert-butyl, pentyl and the like. More preferred are lower alkyl groups having 1 to 4 carbon atoms such as methyl, ethyl, propyl, 2-propyl, n-butyl, isobutyl or t-butyl groups and the like.
  • the alkyl group may be substituted or unsubstituted, and when substituted, preferred groups are: halogen, C 2 -C 6 alkenyl, C 6 -C 10 aryl, C 5 -C 10 heteroaryl, halogen C 1 -C 6 alkyl, 4 to 8 membered heteroalicyclic, hydroxy, C 1 -C 6 alkoxy, C 6 -C 10 aryloxy.
  • Cycloalkyl means a 3 to 8 membered all-carbon monocyclic, all-carbon 5/6 or 6/6-membered fused or polycyclic fused ring ("thickened” ring means each in the system The ring shares an adjacent pair of carbon atoms) groups with other rings in the system, wherein one or more of the rings have a fully linked pi-electron system, and examples of cycloalkyl groups (not limited to) are cyclopropane, cyclobutane, Cyclopentane, cyclopentene, cyclohexane, adamantane, cyclohexadiene, cycloheptane and cycloheptatriene.
  • Cycloalkyl groups are substitutable and substituted.
  • the substituent is preferably one or more groups each selected from the group consisting of: hydrogen, hydroxy, thiol, oxo, lower alkyl, lower alkoxy, lower cycloalkyl, lower heteroalicyclic , lower haloalkoxy, alkylthio, halogen, lower haloalkyl, lower hydroxyalkyl, lower cycloalkylalkylene, lower heteroalicyclic alkylene, aryl, heteroaryl, alkoxycarbonyl, Amino, alkylamino, alkylsulfonyl, arylsulfonyl, alkylaminosulfonyl, arylaminosulfonyl, alkylsulfonylamino, arylsulfonylamino, alkylaminocarbonyl, arylaminocarbonyl, Alkylcarbonylamino, arylcarbonylamino,
  • Heterocycloalkyl means a monocyclic or fused ring radical having from 5 to 9 ring atoms in the ring wherein one or both of the ring atoms are selected from N, O or S(O) p (where p is A hetero atom of 0 to 2 integers, and the remaining ring atoms are C. These rings may have one or more double bonds, but these rings do not have a fully conjugated pi-electron system.
  • Non-limiting examples of unsubstituted heteroalicyclic groups are pyrrolidinyl, piperidino, piperazino, morpholino, thiomorpholino, homopiperazino and the like.
  • the heteroalicyclic group can be substituted or unsubstituted.
  • the substituent is preferably one or more, more preferably one, two or three, and even more preferably one or two, said substituent being selected from the group consisting of hydrogen, hydroxy, thiol, oxygen a lower alkyl group, a lower alkoxy group, a lower cycloalkyl group, a lower heteroalicyclic group, a lower haloalkoxy group, an alkylthio group, a halogen, a lower haloalkyl group, a lower hydroxyalkyl group, a lower cycloalkylalkylene group, Lower heteroalicyclic alkylene, aryl, heteroaryl, alkoxycarbonyl, amino, alkylamino, alkylsulfonyl, arylsulfonyl, alkylaminosulfonyl, arylaminosulfonyl, alkyl Sulfonylamino, arylsulfonylamino, alkylaminocarbony
  • heteroalicyclic groups include, but are not limited to, morpholinyl, piperazinyl, piperidinyl, azetidinyl, pyrrolidinyl, hexahydroazepine, oxetanyl, tetrahydrofuranyl , tetrahydrothiophenyl, oxazolidinyl, thiazolidinyl, isoxazolidinyl, tetrahydropyranyl, thiomorphinyl, quinuclidinyl and imidazolinyl, each group as described above, Examples may also be bicyclic, such as, for example, 3,8-diaza-bicyclo[3.2.1]octane, 2,5-diazabicyclo[2.2.2]octane or octahydro-pyrazine. [2,1-c][1,4]oxazine. Its heteroalicyclic group (and derivatives).
  • Haldroxy means an -OH group.
  • Halogen means fluoro, chloro, bromo or iodo, preferably fluoro or chloro.
  • Amino means a -NH 2 group.
  • substituted by one or more groups means that one, two, three or four hydrogen atoms in a given atom or group are each selected from a specified range of groups. Replace the same or different groups.
  • “Pharmaceutically acceptable salt” means those salts which retain the biological effectiveness and properties of the parent compound. Such salts include:
  • a salt with an acid obtained by a reaction of a free base of a parent compound with an inorganic or organic acid includes hydrochloric acid, hydrobromic acid, nitric acid, phosphoric acid, metaphosphoric acid, sulfuric acid, sulfurous acid, perchloric acid, etc.
  • Organic acids include acetic acid, propionic acid, acrylic acid, oxalic acid, (D) or (L) malic acid, fumaric acid, maleic acid, hydroxybenzoic acid, ⁇ -hydroxybutyric acid, methoxybenzoic acid, ortho-benzene Formic acid, methanesulfonic acid, ethanesulfonic acid, naphthalene-1-sulfonic acid, naphthalene-2-sulfonic acid, p-toluenesulfonic acid, salicylic acid, tartaric acid, citric acid, lactic acid, mandelic acid, succinic acid or malonic acid .
  • an organic base such as ethanolamine, diethanolamine, or the like. Ethanolamine, tromethamine, N-methylglucamine, and the like.
  • “Pharmaceutical composition” means that one or more of the compounds of the invention, or a pharmaceutically acceptable salt, solvate, hydrate or prodrug thereof, is mixed with another chemical component, such as a pharmaceutically acceptable carrier. .
  • the purpose of the pharmaceutical composition is to facilitate the administration to the animal.
  • “Pharmaceutically acceptable carrier” refers to an inactive ingredient in a pharmaceutical composition that does not cause significant irritation to the organism and does not interfere with the biological activity and properties of the administered compound, such as, but not limited to, calcium carbonate, calcium phosphate, various Sugar (eg lactose, mannitol, etc.), starch, cyclodextrin, magnesium stearate, cellulose, magnesium carbonate, acrylic polymer or methacrylic acid polymer, gel, water, polyethylene glycol, propylene glycol, B Glycol, castor oil or hydrogenated castor oil or polyethoxylated hydrogenated castor oil, sesame oil, corn oil, peanut oil and the like.
  • the aforementioned pharmaceutical composition may include, in addition to a pharmaceutically acceptable carrier, an adjuvant which is commonly used in medicine, for example, an antibacterial agent, an antifungal agent, an antimicrobial agent, a quality agent, a tone.
  • an adjuvant which is commonly used in medicine, for example, an antibacterial agent, an antifungal agent, an antimicrobial agent, a quality agent, a tone.
  • Tin reagent (Compound B, 1.0 g, 1.57 mmol), 4-chloro-7-((2-(trimethylsilyl)ethoxy)methyl)-7H--pyrrolo[2,3- d]pyrimidine (Compound A, 540 mg, 1.90 mmol), tetrakis(triphenylphosphine)palladium (183 mg, 0.16 mmol), CuI (0.031 g, 0.16 mmol) in anhydrous DMF (10 mL). The reaction was carried out at ° C for 14 hours. After the reaction was completed, it was poured into water (10 mL), ethyl acetate (100 mL*3), and the organic phase was dried over anhydrous magnesium sulfate. 1) 0.7 g of the target compound was obtained in a yield of 75%.
  • Diisopropylamine was added dropwise to butyllithium (160 mL, 0.4 mol, 10 eq) at 0 ° C, stirred for 1 h, cooled to -78 ° C, and cyclohexanone (40.0 g, 0.4 mol, 10 eq) was added dropwise. Stir for 2 hours with heat.
  • a solution of Compound A (12.0 g, 0.04 mol, 1.0 eq) in THF (100 mL) was added dropwise, and the mixture was warmed to room temperature and stirred for 15 h. Add saturated ammonium chloride solution (50 mL) to the reaction solution, extract with EA (100 mL * 3), combined organic phase spin directly into the next step.
  • the Km value of the different adenosine triphosphate (ATP) concentrations was examined by the Caliper Mobility Shift Assay method to examine the effects of the following enumerated compounds of the invention on inhibitors of the Janus series of kinases.
  • the inhibitory effect of the compounds of the invention on the JAK-STAT pathway was found to be very pronounced.
  • the inhibition of IL-3 and IL-4 induced proliferation of TF-1 cells by the compounds was examined by the MTS method, and the IC 50 value was calculated.
  • TF-1 cells were resuspended in 10 ng/ml IL-3, IL-4 RPMI-1640 medium (containing 10% FBS), and inserted into 96 wells at a density of 15000 cells/well. In the board. Two control groups were set up in the experiment. Negative control group: containing cells, but not containing IL-3 or IL-4 factor, containing 10% serum; positive control group (compound 0 concentration well): cells without compound action, but containing IL-3 or IL-4 Child and serum. Gradiently diluted compounds were added to the cells to a final concentration of 20,000 nM, diluted 3 times, a total of 10 concentrations (containing 0 concentration wells), 3 replicates per concentration.
  • the DMSO content in the medium was 0.1%.
  • the cells were cultured in an incubator at 37 ° C, 5% CO 2 , and the compounds were allowed to act on the cells for 72 h.
  • CCK8 was added, containing 10% (V/V) per well, and incubated at 37 °C for 3 h.
  • the microplate reader detects the absorbance at 450 nM.
  • the compound of the present invention inhibits the action of rheumatoid arthritis, selects DBA/1J mice, and 50 ug of bovine type II collagen with an equal volume of complete Freund's
  • the agent (CFA) was completely emulsified and injected subcutaneously. After 21 days, 50 ug of the same antigen and incomplete Freund's adjuvant (IFA fully emulsified, boosted once. The observation was started from the 45th day. Using 1-4 scoring method: 1 point, normal; 2 points, 1 Joint swelling; 3 points, more than 1 joint swelling, but did not accumulate all joints; 4 points, the entire paw was severely swollen or stiff. The score of each claw was added to obtain the total score of joint inflammation in mice.
  • mice greater than 1 were successfully established in the model. After successfully establishing a mouse model of rheumatoid arthritis, the mice of the present invention were intragastrically administered with the compound of the present invention, and the joint inflammation of the mice was scored after 2 weeks of administration, and the result showed that the product was displayed. It has obvious therapeutic effects on rheumatoid arthritis in mice.

Abstract

Provided are a 4-substituted pyrrolo[2,3-d]pyrimidine compound and the use thereof in preparing medicaments for treating JAK-targeted diseases, such as rheumatoid, immune system diseases, tumours, etc. The 4-substituted pyrrolo[2,3-d]pyrimidine compound is as shown in the structural formula (I). The activity results show that some of compounds as shown in formula (I) have a strong inhibiting effect on Janus kinases.

Description

4-取代吡咯并[2,3-d]嘧啶化合物及其用途4-substituted pyrrolo[2,3-d]pyrimidine compound and use thereof 技术领域Technical field
本发明涉及一种4-取代吡咯并[2,3-d]嘧啶化合物，及其在制备免疫***疾病、治疗类风湿、肿瘤等JAK相关靶点疾病的药物中的用途。The present invention relates to a 4-substituted pyrrolo[2,3-d]pyrimidine compound and its use in the preparation of a medicament for the treatment of diseases of the immune system, treatment of rheumatoid diseases such as rheumatoid, tumors and the like.
背景技术Background technique
蛋白激酶(Protein kinases)又称蛋白质磷酸化酶(Protein phosphakinase)，是一类催化蛋白质磷酸化反应的酶。它能把腺苷三磷酸(ATP)上的γ-磷酸转移到蛋白质分子的氨基酸残基上，从而改变蛋白质、酶的构象和活性。蛋白质的磷酸化是多种信号传导途径的重要环节，细胞内大部分重要的生命活过程都离不开蛋白质磷酸化。这些酶在调节细胞信号包括细胞增殖和细胞分化中是关键的因素。Protein kinases, also known as protein phosphakinase, are a class of enzymes that catalyze the phosphorylation of proteins. It can transfer the γ-phosphate on adenosine triphosphate (ATP) to the amino acid residues of protein molecules, thereby changing the conformation and activity of proteins and enzymes. Phosphorylation of proteins is an important part of many signaling pathways, and most important life processes in cells are inseparable from protein phosphorylation. These enzymes are key factors in regulating cellular signaling, including cell proliferation and cell differentiation.
蛋白激酶信号在转导中主要有两个方面的作用：一是通过磷酸化调节蛋白质的活性，磷酸化和去磷酸化是大多数信号通路组分可逆激活的共同机制，有些蛋白质在磷酸化后具有活性，有些则在去磷酸化后具有活性；二是通过蛋白质的逐级磷酸化，使信号逐级放大，引起细胞反应。Protein kinase signaling plays a major role in transduction: one is to regulate protein activity through phosphorylation. Phosphorylation and dephosphorylation are common mechanisms for reversible activation of most signaling pathway components. Some proteins are phosphorylated. It has activity, some is active after dephosphorylation; the other is to gradually amplify the signal through the stepwise phosphorylation of the protein, causing cell reaction.
Janus激酶\信号传导及转录激活因子(Janus-activated kinase Singal transducers and activators of transcriprion，JAK-STAT)是近年来新发现的一条与细胞因子密切相关的细胞内信号传导通路，参与细胞的增殖、分化、凋亡以及免疫调节等许多重要的生物学过程。Janus激酶是一种非受体型酪氨酸蛋白激酶。有4个家族成员，分别是JAK1、JAK2、TYK2和JAK3。而JAK是一类非常重要的药物靶点，目前JAK抑制剂已经被证实可以用于血液***疾病、肿瘤、类风湿性关节炎及银屑病等治疗药物。由于JAK抑制剂有着显著的医疗用途，可以用于各种相关疾病药物，所以对该类化合物的研究及发现是极其有益的。Janus-activated kinase Singal transducers and activators of transcriprion (JAK-STAT) is a newly discovered intracellular signaling pathway closely related to cytokines, which is involved in cell proliferation and differentiation. Many important biological processes such as apoptosis and immune regulation. Janus kinase is a non-receptor tyrosine protein kinase. There are 4 family members, namely JAK1, JAK2, TYK2 and JAK3. While JAK is a very important drug target, JAK inhibitors have been proven to be useful in the treatment of blood system diseases, tumors, rheumatoid arthritis and psoriasis. Because JAK inhibitors have significant medical uses and can be used in a variety of related disease drugs, the research and discovery of such compounds is extremely beneficial.
中国专利申请CN 102026999A公开了一种氮杂环丁烷和环丁烷衍生物，以及它们的组合物及使用和制备方法，它们是可用于治疗包括例如炎性疾病和自身免疫疾病以及癌症的JAK相关性疾病的JAK抑制剂。其母核结构如下化学式所示。其作为一种有效的JAK抑制剂，可以作为原料药，用于制备治疗类风湿性关节炎、皮肤病、癌症、骨髓增殖性疾病等疾病的药物。Chinese patent application CN 102026999A discloses azetidine and cyclobutane derivatives, as well as compositions and methods of use and preparation thereof, which are useful for the treatment of JAK including, for example, inflammatory and autoimmune diseases and cancer. A JAK inhibitor of a related disease. Its mother core structure is shown in the following chemical formula. As an effective JAK inhibitor, it can be used as a drug substance for the preparation of a medicament for treating diseases such as rheumatoid arthritis, skin diseases, cancer, and myeloproliferative diseases.
Figure PCTCN2015090497-appb-000001
Figure PCTCN2015090497-appb-000001
目前辉瑞公司的Tofacitinib，作为典型的JAK抑制剂药物已经上市，是用于类风湿性关节炎(rheumatoid  arthritis)治疗的首创药物。但是JAK抑制剂的治疗领域涉及面及其广，寻求新的，活性更强，成药性更高的化合物是本领域的一个努力方向。Currently, Pfizer's Tofacitinib, which is marketed as a typical JAK inhibitor, is used for rheumatoid arthritis (rheumatoid). Arthritis) The first drug to treat. However, the therapeutic field of JAK inhibitors involves a wide range of applications, and it is an effort in the field to seek new, more active, and more highly compounded compounds.
发明内容Summary of the invention
本发明的第一技术目的，在于提供一种新的JAK抑制剂化合物；本发明的第二技术目的，在于提供本发明所述JAK抑制剂化合物在制备治疗与JAK抑制剂相关疾病的药物中的应用。A first technical object of the present invention is to provide a novel JAK inhibitor compound; a second technical object of the present invention is to provide a JAK inhibitor compound of the present invention in the preparation of a medicament for treating a disease associated with a JAK inhibitor application.
为了实现本发明的技术目的，本发明的技术方案如下。In order to achieve the technical object of the present invention, the technical solution of the present invention is as follows.
结构如式(I)所示的化合物或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、或其混合物形式、或药学上可接受的盐：a compound of the formula (I) or a tautomer, a mesophil, a racemate, an enantiomer, a diastereomer, or a mixture thereof, or a pharmaceutically acceptable compound Accepted salt:
Figure PCTCN2015090497-appb-000002
Figure PCTCN2015090497-appb-000002
R1选自氢、卤素、烷基。R 1 is selected from the group consisting of hydrogen, halogen, and alkyl.
R2选自：
Figure PCTCN2015090497-appb-000003
Figure PCTCN2015090497-appb-000004
其中R4、R5、R6、R7各自独立的选自氢、卤素、烷基、硝基、氨基、羟基；X选自O、N或S。进一步的R2中R4、R5、R6、R7各自独立的选自氢、卤素、烷基。R 2 is selected from:
Figure PCTCN2015090497-appb-000003
Figure PCTCN2015090497-appb-000004
Wherein R 4 , R 5 , R 6 , and R 7 are each independently selected from the group consisting of hydrogen, halogen, alkyl, nitro, amino, and hydroxy; and X is selected from O, N or S. Further, in R 2 , R 4 , R 5 , R 6 and R 7 are each independently selected from the group consisting of hydrogen, halogen and alkyl.
R3选自：
Figure PCTCN2015090497-appb-000005
Figure PCTCN2015090497-appb-000006
其中R8选自：烷基，环烷基，杂环烷基。 R 3 is selected from:
Figure PCTCN2015090497-appb-000005
Figure PCTCN2015090497-appb-000006
Wherein R 8 is selected from the group consisting of alkyl, cycloalkyl and heterocycloalkyl.
进一步的R3选自
Figure PCTCN2015090497-appb-000007
Further R 3 is selected from
Figure PCTCN2015090497-appb-000007
更进一步的式(I)所示化合物选自：Further compounds of formula (I) are selected from the group consisting of:
Figure PCTCN2015090497-appb-000008
Figure PCTCN2015090497-appb-000008
一种药用组合物，其包含治疗有效量的式(I)中任意一项的化合物、其立体异构体、互变异构体或药学上可接受的盐，以及药学上可接受的载体、辅助剂或溶媒。A pharmaceutical composition comprising a therapeutically effective amount of a compound of any one of formula (I), a stereoisomer, tautomer or pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier , adjuvant or solvent.
更进一步的式(I)所示的化合物、其立体异构体、互变异构体、前药或药学上可接受的盐，其用作药物的用途。进一步的，式(I)所示化合物、其立体异构体、互变异构体、前药或药学上可接受的盐，其用作治疗免疫疾病的药物；用作治疗选自类风湿性关节炎、哮喘、***性红斑狼疮、银屑病、IBD和移植排斥的免疫疾病的药物。Further, a compound represented by the formula (I), a stereoisomer, a tautomer, a prodrug or a pharmaceutically acceptable salt thereof is used as a medicament. Further, a compound of the formula (I), a stereoisomer, a tautomer, a prodrug or a pharmaceutically acceptable salt thereof, which is used as a medicament for treating an immune disease; and a treatment selected from rheumatoid sex Drugs for arthritis, asthma, systemic lupus erythematosus, psoriasis, IBD, and transplant rejection of immune diseases.
更进一步的式(I)所示化合物、其立体异构体、互变异构体、前药或药学上可接受的盐在制备治疗对 JAK1激酶活性的抑制有反应的疾病的药物中的用途，所述疾病特别是免疫疾病，更特别是选自类风湿性关节炎、哮喘、***性红斑狼疮、银屑病、IBD和移植排斥的免疫疾病。Further compounds of the formula (I), stereoisomers, tautomers, prodrugs or pharmaceutically acceptable salts thereof, in the preparation of a therapeutic pair Use of a JAK1 kinase activity for inhibiting a responsive disease, particularly an immune disease, more particularly selected from the group consisting of rheumatoid arthritis, asthma, systemic lupus erythematosus, psoriasis, IBD, and transplant rejection Immune disease.
发明的详细说明Detailed description of the invention
除非有相反陈述，下列用在说明书和权利要求中的术语具有下述含义。Unless otherwise stated, the following terms used in the specification and claims have the following meanings.
“烷基”指饱和的脂族烃基团。包括1至20个碳原子的直链或支链基团。优选含有1至6个碳原子的中等大小烷基，例如甲基、乙基、丙基、2-丙基、正丁基、异丁基、叔丁基、戊基等。更优选的是含有1至4个碳原子的低级烷基，例如甲基、乙基、丙基、2-丙基、正丁基、异丁基或叔丁基等。烷基可以是取代的或未取代的，当被取代时，优选的基团为：卤素、C2-C6烯基、C6-C10芳基、C5-C10杂芳基、卤代C1-C6烷基、4至8元杂脂环基、羟基、C1-C6烷氧基、C6-C10芳氧基。"Alkyl" means a saturated aliphatic hydrocarbon group. A straight or branched chain group of 1 to 20 carbon atoms is included. Preference is given to medium-sized alkyl groups having 1 to 6 carbon atoms, such as methyl, ethyl, propyl, 2-propyl, n-butyl, isobutyl, tert-butyl, pentyl and the like. More preferred are lower alkyl groups having 1 to 4 carbon atoms such as methyl, ethyl, propyl, 2-propyl, n-butyl, isobutyl or t-butyl groups and the like. The alkyl group may be substituted or unsubstituted, and when substituted, preferred groups are: halogen, C 2 -C 6 alkenyl, C 6 -C 10 aryl, C 5 -C 10 heteroaryl, halogen C 1 -C 6 alkyl, 4 to 8 membered heteroalicyclic, hydroxy, C 1 -C 6 alkoxy, C 6 -C 10 aryloxy.
“环烷基”指3至8元全碳单环、全碳5元/6元或6元/6元稠和环或多环稠和环(“稠和”环意味着***中的每个环与***中的其它环共享毗邻的一对碳原子)基团，其中一个或多个环具有完全连接的π电子***，环烷基的实例(不局限于)为环丙烷、环丁烷、环戊烷、环戊烯、环己烷、金刚烷、环己二烯、环庚烷和环庚三烯。环烷基为可取代的和为取代的。当被取代时，取代基优选为一个或多个各自选自以下的基团，包括：氢、羟基、巯基、氧代、低级烷基、低级烷氧基、低级环烷基、低级杂脂环基、低级卤代烷氧基、烷硫基、卤素、低级卤代烷基、低级羟烷基、低级环烷基亚烷基、低级杂脂环基亚烷、芳基、杂芳基、烷氧基羰基、氨基、烷基氨基、烷基磺酰基、芳基磺酰基、烷基氨基磺酰基、芳基氨基磺酰基、烷基磺酰基氨基、芳基磺酰基氨基、烷基氨基羰基、芳基氨基羰基、烷基羰基氨基、芳基羰基氨基。"Cycloalkyl" means a 3 to 8 membered all-carbon monocyclic, all-carbon 5/6 or 6/6-membered fused or polycyclic fused ring ("thickened" ring means each in the system The ring shares an adjacent pair of carbon atoms) groups with other rings in the system, wherein one or more of the rings have a fully linked pi-electron system, and examples of cycloalkyl groups (not limited to) are cyclopropane, cyclobutane, Cyclopentane, cyclopentene, cyclohexane, adamantane, cyclohexadiene, cycloheptane and cycloheptatriene. Cycloalkyl groups are substitutable and substituted. When substituted, the substituent is preferably one or more groups each selected from the group consisting of: hydrogen, hydroxy, thiol, oxo, lower alkyl, lower alkoxy, lower cycloalkyl, lower heteroalicyclic , lower haloalkoxy, alkylthio, halogen, lower haloalkyl, lower hydroxyalkyl, lower cycloalkylalkylene, lower heteroalicyclic alkylene, aryl, heteroaryl, alkoxycarbonyl, Amino, alkylamino, alkylsulfonyl, arylsulfonyl, alkylaminosulfonyl, arylaminosulfonyl, alkylsulfonylamino, arylsulfonylamino, alkylaminocarbonyl, arylaminocarbonyl, Alkylcarbonylamino, arylcarbonylamino.
“杂环烷基”表示单环或稠和环基团，在环中具有5到9个环原子，其中一个或两个环原子是选自N、O或S(O)p(其中p是0至2的整数)的杂原子，其余环原子是C。这些环可以具有一条或多条双键，但这些环不具有完全共轭的π电子***。未取代的杂脂环基的非限制性实例有吡咯烷基、哌啶子基、哌嗪子基、吗啉代基、硫代吗啉代基、高哌嗪子基等。杂脂环基可以是取代的或未取代的。当被取代时，取代基优选为一个或多个，更为优选为一个、两个或三个，进而更优选为一个或两个，所述的取代基选自:氢、羟基、巯基、氧代、低级烷基、低级烷氧基、低级环烷基、低级杂脂环基、低级卤代烷氧基、烷硫基、卤素、低级卤代烷基、低级羟烷基、低级环烷基亚烷基、低级杂脂环基亚烷、芳基、杂芳基、烷氧基羰基、氨基、烷基氨基、烷基磺酰基、芳基磺酰基、烷基氨基磺酰基、芳基氨基磺酰基、烷基磺酰基氨基、芳基磺酰基氨基、烷基氨基羰基、芳基氨基羰基、烷基羰基氨基、芳基羰基氨基。除非另外指出。杂脂环基的实例包括但不限于，吗啉基，哌嗪基，哌啶基，氮杂环丁烷基，吡咯烷基，六氢氮杂□基，氧杂环丁烷基，四氢呋喃基，四氢噻吩基，噁唑烷基，噻唑烷基，异噁唑烷基，四氢吡喃基，硫代吗琳基，奎宁环基和咪唑啉基，各基团如前所述，实例还可以是双环的，诸如，例如，3，8-二氮杂-双环[3.2.1]辛烷、2，5-二氮杂双环[2.2.2]辛烷或八氢-吡嗪并[2，1-c][1,4]噁嗪。其杂脂环基(和衍生物)包括其离子形式。 "Heterocycloalkyl" means a monocyclic or fused ring radical having from 5 to 9 ring atoms in the ring wherein one or both of the ring atoms are selected from N, O or S(O) p (where p is A hetero atom of 0 to 2 integers, and the remaining ring atoms are C. These rings may have one or more double bonds, but these rings do not have a fully conjugated pi-electron system. Non-limiting examples of unsubstituted heteroalicyclic groups are pyrrolidinyl, piperidino, piperazino, morpholino, thiomorpholino, homopiperazino and the like. The heteroalicyclic group can be substituted or unsubstituted. When substituted, the substituent is preferably one or more, more preferably one, two or three, and even more preferably one or two, said substituent being selected from the group consisting of hydrogen, hydroxy, thiol, oxygen a lower alkyl group, a lower alkoxy group, a lower cycloalkyl group, a lower heteroalicyclic group, a lower haloalkoxy group, an alkylthio group, a halogen, a lower haloalkyl group, a lower hydroxyalkyl group, a lower cycloalkylalkylene group, Lower heteroalicyclic alkylene, aryl, heteroaryl, alkoxycarbonyl, amino, alkylamino, alkylsulfonyl, arylsulfonyl, alkylaminosulfonyl, arylaminosulfonyl, alkyl Sulfonylamino, arylsulfonylamino, alkylaminocarbonyl, arylaminocarbonyl, alkylcarbonylamino, arylcarbonylamino. Unless otherwise stated. Examples of heteroalicyclic groups include, but are not limited to, morpholinyl, piperazinyl, piperidinyl, azetidinyl, pyrrolidinyl, hexahydroazepine, oxetanyl, tetrahydrofuranyl , tetrahydrothiophenyl, oxazolidinyl, thiazolidinyl, isoxazolidinyl, tetrahydropyranyl, thiomorphinyl, quinuclidinyl and imidazolinyl, each group as described above, Examples may also be bicyclic, such as, for example, 3,8-diaza-bicyclo[3.2.1]octane, 2,5-diazabicyclo[2.2.2]octane or octahydro-pyrazine. [2,1-c][1,4]oxazine. Its heteroalicyclic group (and derivatives) include its ionic form.
“羟基”表示-OH基团。"Hydroxy" means an -OH group.
“卤素”表示氟、氯、溴或碘，优选为氟或氯。"Halogen" means fluoro, chloro, bromo or iodo, preferably fluoro or chloro.
“氨基”表示-NH2基团。"Amino" means a -NH 2 group.
所谓“任选地”的意思是指后续描述的事件或情形可能会也可能不会发生，并且该描述包括事物或情形可能会也可能不会发生，并且该描述包括事物或情形发生和不发生两种情况。By "optionally" is meant that the subsequently described event or circumstance may or may not occur, and that the description may or may not occur, and that the description includes or does not occur. Two situations.
在一些实施方案中，“被一个或多个基团取代”是指在指定的原子或基团中的一个、两个、三个或四个氢原子分别被指定范围的基团中选出的相同或不同的基团替换。In some embodiments, "substituted by one or more groups" means that one, two, three or four hydrogen atoms in a given atom or group are each selected from a specified range of groups. Replace the same or different groups.
“药学上可接受的盐”表示保留母体化合物的生物有效性和性质的那些盐。这类盐包括："Pharmaceutically acceptable salt" means those salts which retain the biological effectiveness and properties of the parent compound. Such salts include:
(1)与酸成盐，通过母体化合物的游离碱与无机酸或有机酸的反应而得，无机酸包括盐酸、氢溴酸、硝酸、磷酸、偏磷酸、硫酸、亚硫酸和高氯酸等，有机酸包括乙酸、丙酸、丙烯酸、草酸、(D)或(L)苹果酸、富马酸、马来酸、羟基苯甲酸、γ-羟基丁酸、甲氧基苯甲酸、邻苯二甲酸、甲磺酸、乙磺酸、萘-1-磺酸、萘-2-磺酸、对甲苯磺酸、水杨酸、酒石酸、柠檬酸、乳酸、扁桃酸、琥珀酸或丙二酸等。(1) a salt with an acid obtained by a reaction of a free base of a parent compound with an inorganic or organic acid, and the inorganic acid includes hydrochloric acid, hydrobromic acid, nitric acid, phosphoric acid, metaphosphoric acid, sulfuric acid, sulfurous acid, perchloric acid, etc. Organic acids include acetic acid, propionic acid, acrylic acid, oxalic acid, (D) or (L) malic acid, fumaric acid, maleic acid, hydroxybenzoic acid, γ-hydroxybutyric acid, methoxybenzoic acid, ortho-benzene Formic acid, methanesulfonic acid, ethanesulfonic acid, naphthalene-1-sulfonic acid, naphthalene-2-sulfonic acid, p-toluenesulfonic acid, salicylic acid, tartaric acid, citric acid, lactic acid, mandelic acid, succinic acid or malonic acid .
(2)存在于母体化合物中的酸性质子被金属离子代替或者与有机碱配位化合所生成的盐，金属例子例如碱金属离子、碱土金属离子或铝离子，有机碱例如乙醇胺、二乙醇胺、三乙醇胺、氨丁三醇、N-甲基葡糖胺等。(2) a salt formed by replacing an acidic proton in a parent compound with a metal ion or by complexing with an organic base, such as an alkali metal ion, an alkaline earth metal ion or an aluminum ion, an organic base such as ethanolamine, diethanolamine, or the like. Ethanolamine, tromethamine, N-methylglucamine, and the like.
“药物组合物”指将本发明中的化合物中的一个或多个或其药学上可接受的盐、溶剂化物、水合物或前药与别的化学成分，例如药学上可接受的载体，混合。药物组合物的目的是促进给药给动物的过程。"Pharmaceutical composition" means that one or more of the compounds of the invention, or a pharmaceutically acceptable salt, solvate, hydrate or prodrug thereof, is mixed with another chemical component, such as a pharmaceutically acceptable carrier. . The purpose of the pharmaceutical composition is to facilitate the administration to the animal.
“药用载体”指的是对有机体不引起明显的刺激性和不干扰所给予化合物的生物活性和性质的药物组合物中的非活性成分，例如但不限于：碳酸钙、磷酸钙、各种糖(例如乳糖、甘露醇等)、淀粉、环糊精、硬脂酸镁、纤维素、碳酸镁、丙烯酸聚合物或甲基丙烯酸聚合物、凝胶、水、聚乙二醇、丙二醇、乙二醇、蓖麻油或氢化蓖麻油或多乙氧基氢化蓖麻油、芝麻油、玉米油、花生油等。"Pharmaceutically acceptable carrier" refers to an inactive ingredient in a pharmaceutical composition that does not cause significant irritation to the organism and does not interfere with the biological activity and properties of the administered compound, such as, but not limited to, calcium carbonate, calcium phosphate, various Sugar (eg lactose, mannitol, etc.), starch, cyclodextrin, magnesium stearate, cellulose, magnesium carbonate, acrylic polymer or methacrylic acid polymer, gel, water, polyethylene glycol, propylene glycol, B Glycol, castor oil or hydrogenated castor oil or polyethoxylated hydrogenated castor oil, sesame oil, corn oil, peanut oil and the like.
前述的药物组合物中，除了包括药学上可接受的载体外，还可以包括在药(剂)学上常用的辅剂，例如：抗细菌剂、抗真菌剂、抗微生物剂、保质剂、调色剂、增溶剂、增稠剂、表面活性剂、络合剂、蛋白质、氨基酸、脂肪、糖类、维生素、矿物质、微量元素、甜味剂、色素、香精或它们的结合等。The aforementioned pharmaceutical composition may include, in addition to a pharmaceutically acceptable carrier, an adjuvant which is commonly used in medicine, for example, an antibacterial agent, an antifungal agent, an antimicrobial agent, a quality agent, a tone. Colorants, solubilizers, thickeners, surfactants, complexing agents, proteins, amino acids, fats, sugars, vitamins, minerals, trace elements, sweeteners, colors, flavors or combinations thereof.
具体实施方式detailed description
实施例1：2-(3-(7H,7'H-[4,5'-联吡咯[2,3-d]嘧啶]-7'-基)-1-(乙基磺酰基)氮杂环丁烷-3-基)乙腈(化合物1)的制备 Example 1: 2-(3-(7H,7'H-[4,5'-bipyrrole[2,3-d]pyrimidin]-7'-yl)-1-(ethylsulfonyl)aza Preparation of cyclobutane-3-yl)acetonitrile (compound 1)
Figure PCTCN2015090497-appb-000009
Figure PCTCN2015090497-appb-000009
合成路线：synthetic route:
Figure PCTCN2015090497-appb-000010
1)7-((2-(三甲基硅基)乙氧基)甲基)-7H,7'H-4,5'-联吡咯[2,3-d]嘧啶(化合物C)的制备
Figure PCTCN2015090497-appb-000010
1) Preparation of 7-((2-(trimethylsilyl)ethoxy)methyl)-7H,7'H-4,5'-bipyrrole[2,3-d]pyrimidine (Compound C)
将4-氯-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H--吡咯并[2,3-d]嘧啶(化合物A，500mg，1.25mmol，1.0eq)和5-(4,4,5,5-四甲基-1,3,2-二氧硼杂戊环-2-基)-7-甲苯磺酰基-7H-吡咯并[2,3-d]嘧啶(化合物B，392mg，1.38mmol，1.1eq)溶于DME(10mL)/2M碳酸钠溶液(5mL)的混合液中，N2保护下加入Pd(PPh3)4(144mg，0.125mmol，0.1eq)，加完后升温至回流反应3h。TLC监测反应完全(Rf＝0.1，PE：EA＝3:1)，冷却至室温后加入H2O(30mL)和EA(50mL)分液，水相用EA萃取三次后，合并有机相，饱和盐水洗涤一次，有机相经无水硫酸镁干燥后，柱层析(PE:EA＝3:1-1:1)得淡黄色固体224mg(化合物C)。4-Chloro-7-((2-(trimethylsilyl)ethoxy)methyl)-7H--pyrrolo[2,3-d]pyrimidine (Compound A, 500 mg, 1.25 mmol, 1.0 Eq) and 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-7-toluenesulfonyl-7H-pyrrolo[2,3 mixture / 2M sodium carbonate solution (5mL) of -d] pyrimidine (compound B, 392mg, 1.38mmol, 1.1eq) was dissolved in DME (10mL) was added Pd (PPh 3) 4 (144mg under N 2, 0.125 Methyl, 0.1 eq), after the addition was completed, the temperature was raised to reflux for 3 h. The reaction was monitored by TLC (Rf=0.1, PE: EA=3:1). After cooling to room temperature, H 2 O (30 mL) and EA (50 mL) were added and the aqueous phase was extracted three times with EA. After washing with brine, the organic layer was dried (MgSO4)
2)2-(1-(乙基磺酰基)-3-(7-((2-(三甲基硅基)乙氧基)甲基)-7H,7’H-[4,5'-联吡咯[2,3-d]嘧啶]-7'-基)氮杂环丁烷-3-基)乙腈(化合物D)的制备2) 2-(1-(ethylsulfonyl)-3-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H,7'H-[4,5'- Preparation of bipyrrole [2,3-d]pyrimidin]-7'-yl)azetidin-3-yl)acetonitrile (Compound D)
将7-((2-(三甲基硅基)乙氧基)甲基)-7H,7'H-4,5'-联吡咯[2,3-d]嘧啶(化合物C，224mg，0.61mmol，1.0eq)溶于乙腈(5mL)中，加入2-(1-(乙基磺酰基)氮杂环丁烷-3-亚基)乙腈(136mg，0.73mmol，1.2eq)后滴加DBU(111mg，0.73mmol，1.2eq)，36℃反应20h，TLC检测未反应完，补加2-(1-(乙基磺酰基)氮杂环丁烷-3-亚基)乙腈(136mg，0.73mmol，1.2eq)，DBU(111mg，0.73mmol，1.2eq)，加完32℃反应20h。TLC监测反应(Rf＝0.2，PE：EA＝1:1)原料未反应完，旋干溶剂，柱层析(PE:EA＝1:1)得黄色油状物化合物D 107mg，收率：32％。7-((2-(Trimethylsilyl)ethoxy)methyl)-7H,7'H-4,5'-bipyrrole[2,3-d]pyrimidine (Compound C, 224 mg, 0.61 Methyl acetate (5 eq.) was dissolved in acetonitrile (5 mL). EtOAc (EtOAc, EtOAc (EtOAc) (111 mg, 0.73 mmol, 1.2 eq), reacted at 36 ° C for 20 h, TLC detection was not reacted, and added 2-(1-(ethylsulfonyl)azetidin-3-ylidene)acetonitrile (136 mg, 0.73) Methyl, 1.2 eq., DBU (111 mg, 0.73 mmol, 1.2 eq). TLC monitoring reaction (Rf = 0.2, PE: EA = 1:1). The starting material was not reacted, the solvent was evaporated, and the column chromatography (PE: EA = 1:1) gave a yellow oily compound D 107 mg, yield: 32% .
3)2-(3-(7H,7'H-[4,5'-联吡咯[2,3-d]嘧啶]-7'-基)-1-(乙基磺酰基)氮杂环丁烷-3-基)乙腈(化合物1)的制备3) 2-(3-(7H,7'H-[4,5'-bipyrrole[2,3-d]pyrimidin]-7'-yl)-1-(ethylsulfonyl)azetidin Preparation of alk-3-yl)acetonitrile (Compound 1)
将2-(1-(乙基磺酰基)-3-(7-((2-(三甲基硅基)乙氧基)甲基)-7H,7’H-[4,5'-联吡咯[2,3-d]嘧啶]-7'-基)氮杂环丁烷-3-基)乙腈(化合物D，107mg，0.19mmol，1.0eq)溶于乙腈(10mL)中，加入三氟化硼***溶液(68mg，0.48mmol，2.5eq)后30℃反应20h，加入水(1.2mL)搅拌3h后，滴加氨水(2mL) 和水(3mL)后搅拌反应过夜。LC-MS检测反应结束，旋干溶剂直接柱层析得黄色固体60mg，加入10mL甲醇超声过滤的白色固体(化合物1)17mg，收率：21.3％。2-(1-(ethylsulfonyl)-3-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H,7'H-[4,5'-linked Pyrrole [2,3-d]pyrimidin-7-yl)azetidin-3-yl)acetonitrile (Compound D, 107 mg, 0.19 mmol, 1.0 eq) was dissolved in EtOAc (10 mL) The solution of boron ether (68 mg, 0.48 mmol, 2.5 eq) was reacted at 30 ° C for 20 h, added with water (1.2 mL), stirred for 3 h, then aqueous ammonia (2 mL) was added dropwise. After stirring with water (3 mL), the reaction was stirred overnight. After completion of the reaction by LC-MS, the solvent was directly purified by column chromatography to give a yellow solid (yield: 60 mg), and a white solid (Comp.
HPLC:98.09％；MS(ESI)m/z:[M+H]+＝423.1；1H-NMR(400 MHz,DMSO-d6)δ:12.22(s,1H),9.98(s,1H),8.96(s,1H),8.87(s,1H),8.64(s,1H),7.69(t,1H),7.26(d,1H),4.87(d,2H),4.40(d,2H),3.79(s,2H),3.25(d,2H),1.25(t,3H)ppm。HPLC: 98.09%; MS (ESI) m/z: [M+H] + =423.1; 1 H-NMR (400 MHz, DMSO-d6) δ: 12.22 (s, 1H), 9.98 (s, 1H), 8.96 (s, 1H), 8.87 (s, 1H), 8.64 (s, 1H), 7.69 (t, 1H), 7.26 (d, 1H), 4.87 (d, 2H), 4.40 (d, 2H), 3.79 (s, 2H), 3.25 (d, 2H), 1.25 (t, 3H) ppm.
实施例2：2-(3-(3-(7H-吡咯[2,3-d]嘧啶-4-基)-1H-吡咯[2,3-c]吡啶-1-基)-1-(乙基磺酰基)氮杂环丁烷-3-基)乙腈(化合物2)的制备Example 2: 2-(3-(3-(7H-pyrrole[2,3-d]pyrimidin-4-yl)-1H-pyrrole[2,3-c]pyridin-1-yl)-1-( Preparation of ethylsulfonyl)azetidin-3-yl)acetonitrile (Compound 2)
Figure PCTCN2015090497-appb-000011
Figure PCTCN2015090497-appb-000011
参考实施例1的合成方法，制备了化合物2 51mg，收率：24.7％。With reference to the synthesis method of Example 1, 51 mg of Compound 2 was prepared in a yield: 24.7%.
HPLC:99.68％；MS(ESI)m/z:[M+H]+＝422.6；1H-NMR(400MHz,DMSO-d6)δ:12.16(s,1H),8.83(d,2H),8.63(d,1H),8.53(s,1H),8.41(d,1H),7.64(d,1H),7.16(d,1H),4.83(d,2H),4.51(d,2H),3.72(s,2H),3.24(d,2H),1.25(t,3H)ppm。HPLC: 99.68%; MS (ESI) m/z: [M+H] + = 422.6; 1 H-NMR (400 MHz, DMSO-d6) δ: 12.16 (s, 1H), 8.83 (d, 2H), 8.63 (d, 1H), 8.53 (s, 1H), 8.41 (d, 1H), 7.64 (d, 1H), 7.16 (d, 1H), 4.83 (d, 2H), 4.51 (d, 2H), 3.72 ( s, 2H), 3.24 (d, 2H), 1.25 (t, 3H) ppm.
实施例3：2-(3-(3-(7H-吡咯[2,3-d]嘧啶-4-基)-1H-吡咯[2,3-b]吡啶-1-基)-1-(乙基磺酰基)氮杂环丁烷-3-基)乙腈(化合物3)的制备Example 3: 2-(3-(3-(7H-pyrrole[2,3-d]pyrimidin-4-yl)-1H-pyrrole[2,3-b]pyridin-1-yl)-1-( Preparation of ethylsulfonyl)azetidin-3-yl)acetonitrile (compound 3)
Figure PCTCN2015090497-appb-000012
Figure PCTCN2015090497-appb-000012
参考实施例1的合成方法，制备了化合物3 64mg，收率：29.4％。With reference to the synthesis method of Example 1, 64 mg of Compound 3 was prepared in a yield: 29.4%.
HPLC:97.0％；MS(ESI)m/z:[M+H]+＝422.1。HPLC: 97.0%; MS (ESI ) m / z: [M + H] + = 422.1.
实施例4：2-(3-(3-(7H-吡咯[2,3-d]嘧啶-4-基)-1H-吡咯[3,2-c]吡啶-1-基)-1-(乙基磺酰基)氮杂环丁烷-3-基)乙腈(化合物4)的制备 Example 4: 2-(3-(3-(7H-pyrrole[2,3-d]pyrimidin-4-yl)-1H-pyrrole[3,2-c]pyridin-1-yl)-1-( Preparation of ethylsulfonyl)azetidin-3-yl)acetonitrile (Compound 4)
Figure PCTCN2015090497-appb-000013
Figure PCTCN2015090497-appb-000013
参考实施例1的合成方法，制备了化合物4 14mg，收率：19.3％。With reference to the synthesis method of Example 1, 14 mg of Compound 4 was prepared in a yield: 19.3%.
HPLC:99.62％；MS(ESI)m/z:[M+H]+＝422.6；1H-NMR(400MHz,DMSO-d6)δ:12.17(s,1H),9.97(s,1H),8.86(s,1H),8.39(m,2H),7.65(s,1H),7.48(d,1H),7.18(s,1H),4.76(d,2H),4.52(d,2H),3.67(s,2H),3.25(d,2H),1.25(t,3H)ppm。HPLC: 99.62%; MS (ESI) m/z: [M+H] + = 422.6; 1 H-NMR (400 MHz, DMSO-d6) δ: 12.17 (s, 1H), 9.97 (s, 1H), 8.86 (s, 1H), 8.39 (m, 2H), 7.65 (s, 1H), 7.48 (d, 1H), 7.18 (s, 1H), 4.76 (d, 2H), 4.52 (d, 2H), 3.67 ( s, 2H), 3.25 (d, 2H), 1.25 (t, 3H) ppm.
实施例5：2-(3-(3-(7H-吡咯[2,3-d]嘧啶-4-基)-1H-吡咯[3,2-b]吡啶-1-基)-1-(乙基磺酰基)氮杂环丁烷-3-基)乙腈(化合物5)的制备Example 5: 2-(3-(3-(7H-pyrrole[2,3-d]pyrimidin-4-yl)-1H-pyrrole[3,2-b]pyridin-1-yl)-1-( Preparation of ethylsulfonyl)azetidin-3-yl)acetonitrile (compound 5)
Figure PCTCN2015090497-appb-000014
Figure PCTCN2015090497-appb-000014
参考实施例1的合成方法，制备了化合物5 36mg，收率：32.1％。With reference to the synthesis method of Example 1, Compound 5 36 mg was obtained in a yield: 32.1%.
HPLC:97.47％；MS(ESI)m/z:[M+H]+＝422.5；1H-NMR(400MHz,DMSO-d6)δ:11.99(s,1H),8.74(s,1H),8.64(d,1H),8.59(s,1H),7.91(d,1H),7.58(s,1H),7.51(s,1H),7.33(m,1H),4.72(d,2H),4.46(d,2H),3.67(s,2H),3.18(m,2H),1.24(t,3H)ppm。HPLC: 97.47%; MS (ESI) m/z: [M+H] + =422.5; 1 H-NMR (400 MHz, DMSO-d6) δ: 11.99 (s, 1H), 8.74 (s, 1H), 8.64 (d, 1H), 8.59 (s, 1H), 7.91 (d, 1H), 7.58 (s, 1H), 7.51 (s, 1H), 7.33 (m, 1H), 4.72 (d, 2H), 4.46 ( d, 2H), 3.67 (s, 2H), 3.18 (m, 2H), 1.24 (t, 3H) ppm.
实施例6：(3-(7H-吡咯并[2,3-d]嘧啶-4-基)-1H-吲哚-1-基)-3-环戊基丙氰(化合物6)的制备Example 6: Preparation of (3-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-indol-1-yl)-3-cyclopentylpropane cyanide (Compound 6)
Figure PCTCN2015090497-appb-000015
Figure PCTCN2015090497-appb-000015
参考实施例1的合成方法，制备了化合物6 130mg，收率：49.1％。With reference to the synthesis method of Example 1, Compound 6 130 mg was obtained in a yield: 49.1%.
1H-NMR(400MHz,DMSO-d6)δ12.04(s，1H)，8.77-8.79(m，2H)，8.53(s，1H)，7.76(d，1H)，7.58(s，1H)，7.21(m，2H)，7.10(s，1H)，4.84(m，1H)，3.56(m，2H)，2.66(m，1H)，1.88(m，1H)，1.69(m，1H)，1.43(m，4H)，1.24(m，1H)，1.01(m，1H)ppm。 1 H-NMR (400MHz, DMSO -d6) δ12.04 (s, 1H), 8.77-8.79 (m, 2H), 8.53 (s, 1H), 7.76 (d, 1H), 7.58 (s, 1H), 7.21 (m, 2H), 7.10 (s, 1H), 4.84 (m, 1H), 3.56 (m, 2H), 2.66 (m, 1H), 1.88 (m, 1H), 1.69 (m, 1H), 1.43 (m, 4H), 1.24 (m, 1H), 1.01 (m, 1 H) ppm.
实施例7：2-(3-(3-(7H-吡咯[2,3-d]嘧啶-4-基)-1H-吲唑-1-基)-1-(乙基磺酰基)氮杂环丁烷-3-基)乙腈(化合物7)的制备Example 7: 2-(3-(3-(7H-pyrrole[2,3-d]pyrimidin-4-yl)-1H-indazol-1-yl)-1-(ethylsulfonyl)aza Preparation of cyclobutane-3-yl)acetonitrile (compound 7)
Figure PCTCN2015090497-appb-000016
Figure PCTCN2015090497-appb-000016
参考实施例1的合成方法，制备了化合物7 65.8mg，收率22％。With reference to the synthesis method of Example 1, 65.8 mg of Compound 7 was prepared in a yield of 22%.
1H-NMR(400MHz,DMSO-d6)δ(ppm)12.34(s,1H),8.94(s,1H),8.86-8.84(d,J＝8.12Hz,1H),7.75-7.73(d,J＝8.52Hz,1H),7.68-7.60(t,1H),7.60-7.56(t,1H),7.46-7.42(t,1H),4.82-4.80(d,J＝8.96Hz,1H),4.55-4.52(d,J＝9.04Hz,1H),3.73(s,2H),3.29-3.23(m,2H),1.26-1.23(t,3H)；LC-MS(ESI+APCI)[M+H]+422.23。 1 H-NMR (400MHz, DMSO -d6) δ (ppm) 12.34 (s, 1H), 8.94 (s, 1H), 8.86-8.84 (d, J = 8.12Hz, 1H), 7.75-7.73 (d, J =8.52 Hz, 1H), 7.68-7.60 (t, 1H), 7.60-7.56 (t, 1H), 7.46-7.42 (t, 1H), 4.82-4.80 (d, J = 8.96 Hz, 1H), 4.55- 4.52 (d, J=9.04 Hz, 1H), 3.73 (s, 2H), 3.29-3.23 (m, 2H), 1.26-1.23 (t, 3H); LC-MS (ESI+APCI) [M+H] + 422.23.
实施例8：2-(3-(3-(7H-吡咯[2,3-d]嘧啶-4-基)-1H-吡咯[3,2-c]吡啶-1-基)-1-(甲基磺酰基)氮杂环丁烷-3-基)乙腈(化合物8)的制备Example 8: 2-(3-(3-(7H-pyrrole[2,3-d]pyrimidin-4-yl)-1H-pyrrole[3,2-c]pyridin-1-yl)-1-( Preparation of Methylsulfonyl)azetidin-3-yl)acetonitrile (Compound 8)
Figure PCTCN2015090497-appb-000017
Figure PCTCN2015090497-appb-000017
参考实施例1的合成方法，制备了化合物8，淡黄色固体，505mg,收率83％。Referring to the synthesis method of Example 1, Compound 8 was obtained as a pale yellow solid, 505 mg, yield 83%.
HPLC：98.13％，MS(ESI)m/z:[M+H]+＝408.2。1H-NMR(400MHz,DMSO-d6)δ:3.10(3H,s),3.67(2H,s),4.40-4.60(2H,m),4.70-4.90(2H,m),7.18(1H,s),7.48(1H,s),7.65(1H,s),8.89(1H,s),8.85(1H,s),9.96(1H,s),12.17(1H,brs)。HPLC: 98.13%, MS (ESI ) m / z: [M + H] + = 408.2. 1 H-NMR (400MHz, DMSO -d6) δ: 3.10 (3H, s), 3.67 (2H, s), 4.40-4.60 (2H, m), 4.70-4.90 (2H, m), 7.18 (1H, s ), 7.48 (1H, s), 7.65 (1H, s), 8.89 (1H, s), 8.85 (1H, s), 9.96 (1H, s), 12.17 (1H, brs).
实施例9：2-(3-(3-(7H-吡咯[2,3-d]嘧啶-4-基)-1H-吡咯[3,2-c]吡啶-1-基)-1-((4-甲基哌嗪-1-基)磺酰基)氮杂环丁烷-3-基)乙腈(化合物9)的制备Example 9: 2-(3-(3-(7H-pyrrole[2,3-d]pyrimidin-4-yl)-1H-pyrrole[3,2-c]pyridin-1-yl)-1-( Preparation of (4-methylpiperazin-1-yl)sulfonyl)azetidin-3-yl)acetonitrile (Compound 9)
Figure PCTCN2015090497-appb-000018
Figure PCTCN2015090497-appb-000018
参考实施例1的合成方法，制备了化合物9，白色固体，210mg，收率59％。Referring to the synthesis method of Example 1, Compound 9, a white solid, 210 mg, yield 59% was obtained.
HPLC：98.76％，MS(ESI)m/z:[M+H]+＝492.3。1H NMR(400MHz,DMSO-d6)δ:2.24(3H,s),2.40-2.50(4H,m),3.20-3.30(4H,m),3.66(2H,s),4.49(2H,d,J＝8.4Hz),4.62(2H,d,J＝8.8Hz),7.18(1H,d,J＝2.0Hz),7.50(1H,d,J＝5.6Hz),7.65(1H,dd,J＝5.6,2.4Hz),8.38(1H,d,J＝5.6Hz),8.44(1H,s),8.86(1H,s),9.97(1H,s),12.18(1H,brs)。HPLC: 98.76%, MS (ESI ) m / z: [M + H] + = 492.3. 1 H NMR (400MHz, DMSO- d6) δ: 2.24 (3H, s), 2.40-2.50 (4H, m), 3.20-3.30 (4H, m), 3.66 (2H, s), 4.49 (2H, d, J = 8.4 Hz), 4.62 (2H, d, J = 8.8 Hz), 7.18 (1H, d, J = 2.0 Hz), 7.50 (1H, d, J = 5.6 Hz), 7.65 (1H, dd, J = 5.6, 2.4 Hz), 8.38 (1H, d, J = 5.6 Hz), 8.44 (1H, s), 8.86 (1H, s), 9.97 (1H, s), 12.18 (1H, brs).
实施例10：3-(7H-吡咯并[2,3-d]嘧啶-4-基)-1H-吡咯[3,2-c]吡啶-1-基)-3-环戊基丙氰(化合物10)的制备Example 10: 3-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrrole[3,2-c]pyridin-1-yl)-3-cyclopentylpropane cyanide ( Preparation of compound 10)
Figure PCTCN2015090497-appb-000019
Figure PCTCN2015090497-appb-000019
参考实施例1的合成方法，制备了化合物10，黄色固体，70mg，收率：11％。Referring to the synthesis method of Example 1, Compound 10 was prepared as a yellow solid, 70 mg, yield: 11%.
HPLC：98.86％，MS(ESI)m/z:[M+H]+＝357.5。1H NMR(400MHz,DMSO-d6)δ:1.00-2.00(9H,m),2.60-2.80(1H,m),3.20-3.30(1H,m,overlapwithwater),3.55-3.65(1H,m),4.80-5.00(1H,m),7.16(1H,s),7.64(1H,s),7.82(1H,d,J＝5.6Hz),8.36(1H,d,J＝5.6Hz),8.63(1H,s),8.84(1H,s),9.95(1H,s),12.14(1H,brs)。HPLC: 98.86%, MS (ESI ) m / z: [M + H] + = 357.5. 1 H NMR (400MHz, DMSO- d6) δ: 1.00-2.00 (9H, m), 2.60-2.80 (1H, m), 3.20-3.30 (1H, m, overlapwithwater), 3.55-3.65 (1H, m), 4.80-5.00 (1H, m), 7.16 (1H, s), 7.64 (1H, s), 7.82 (1H, d, J = 5.6 Hz), 8.36 (1H, d, J = 5.6 Hz), 8.63 (1H) , s), 8.84 (1H, s), 9.95 (1H, s), 12.14 (1H, brs).
实施例11：2-(3-(3-(7H-吡咯[2,3-d]嘧啶-4-基)-4,5,6,7-四氢-1H-吡唑[4,3-c]吡啶-1-基)-1-(乙基磺酰基)氮杂环丁烷-3-基)乙腈(化合物11)的制备Example 11: 2-(3-(3-(7H-pyrrole[2,3-d]pyrimidin-4-yl)-4,5,6,7-tetrahydro-1H-pyrazole [4,3- Preparation of c]pyridin-1-yl)-1-(ethylsulfonyl)azetidin-3-yl)acetonitrile (Compound 11)
Figure PCTCN2015090497-appb-000020
Figure PCTCN2015090497-appb-000020
合成路线： synthetic route:
Figure PCTCN2015090497-appb-000021
Figure PCTCN2015090497-appb-000021
1)叔丁基-1-(4-甲氧基苄基)-3-(7-((2-(三甲基硅基)乙氧基)甲基)-7H-吡咯[2,3-d]嘧啶-4-基)-1,4,6,7-四氢-5H-吡唑[4,3-c]吡啶-5-甲酸酯(化合物C)的制备1) tert-Butyl-1-(4-methoxybenzyl)-3-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrole [2,3- Preparation of d]pyrimidin-4-yl)-1,4,6,7-tetrahydro-5H-pyrazole[4,3-c]pyridine-5-carboxylate (Compound C)
将锡试剂(化合物B，1.0g,1.57mmol)、4-氯-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H--吡咯并[2,3-d]嘧啶(化合物A，540mg，1.90mmol)、四(三苯基膦)钯(183mg，0.16mmol)、CuI(0.031g,0.16mmol)加到无水DMF(10mL)中，氮气氛，80℃反应14小时。反应结束后倒入水(10mL)中，乙酸乙酯(100mL*3)萃取，有机相经无水硫酸镁干燥，制砂后柱层析(洗脱剂：石油醚/乙酸乙酯＝100:1)得目标化合物0.7g，收率75％。Tin reagent (Compound B, 1.0 g, 1.57 mmol), 4-chloro-7-((2-(trimethylsilyl)ethoxy)methyl)-7H--pyrrolo[2,3- d]pyrimidine (Compound A, 540 mg, 1.90 mmol), tetrakis(triphenylphosphine)palladium (183 mg, 0.16 mmol), CuI (0.031 g, 0.16 mmol) in anhydrous DMF (10 mL). The reaction was carried out at ° C for 14 hours. After the reaction was completed, it was poured into water (10 mL), ethyl acetate (100 mL*3), and the organic phase was dried over anhydrous magnesium sulfate. 1) 0.7 g of the target compound was obtained in a yield of 75%.
2)叔丁基-3-(7-((2-(三甲基硅基)乙氧基)甲基)-7H-吡咯[2,3-d]嘧啶-4-基)-1,4,6,7-四氢-5H-吡唑[4,3-c]吡啶-5-甲酸酯(化合物D)的制备2) tert-Butyl-3-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrole[2,3-d]pyrimidin-4-yl)-1,4 Of 6,6-tetrahydro-5H-pyrazole[4,3-c]pyridine-5-carboxylate (Compound D)
将化合物C(0.4g,0.68mmol)、Pd/C(0.1g)加到无水甲醇(10mL)中，氢气氛，30℃反应12小时。产物经柱层析(洗脱剂：石油醚/乙酸乙酯＝50:1)得目标化合物D 0.26g，收率81.3％。Compound C (0.4 g, 0.68 mmol) and Pd/C (0.1 g) were added to anhydrous methanol (10 mL), and the mixture was reacted at 30 ° C for 12 hours. The product was subjected to column chromatography (eluent: petroleum ether / ethyl acetate = 50:1) to give the object compound D 0.26 g.
3)叔丁基-1-(3-(腈甲基)-1-(乙基磺酰基)氮杂环丁烷-3-基)-3-(7-((2-(三甲基硅基)乙氧基)甲基)-7H-吡咯[2,3-d]嘧啶-4-基)-1,4,6,7-四氢-5H-吡唑[4,3-c]吡啶-5-甲酸酯(化合物E)的制备3) tert-Butyl-1-(3-(nitrilemethyl)-1-(ethylsulfonyl)azetidin-3-yl)-3-(7-((2-(trimethylsilane)) Ethyl)methyl)-7H-pyrrole[2,3-d]pyrimidin-4-yl)-1,4,6,7-tetrahydro-5H-pyrazole[4,3-c]pyridine Preparation of 5-formate (Compound E)
将叔丁基-3-(7-((2-(三甲基硅基)乙氧基)甲基)-7H-吡咯[2,3-d]嘧啶-4-基)-1,4,6,7-四氢-5H-吡唑[4,3-c]吡啶-5-甲酸酯(化合物D，0.26g，0.55mmol)溶于乙腈(10mL)中，加入2-(1-(乙基磺酰基)氮杂环丁烷-3-亚基)乙腈(123mg，0.66mmol)，DBU(126mg，0.82mmol)，氮气氛，25℃反应12h，反应结束后经柱层析得黄色固体化合物E 0.3g，收率82.6％。tert-Butyl-3-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrole[2,3-d]pyrimidin-4-yl)-1,4, 6,7-Tetrahydro-5H-pyrazol[4,3-c]pyridine-5-carboxylate (Compound D, 0.26 g, 0.55 mmol) was dissolved in acetonitrile (10 mL). Ethylsulfonyl)azetidin-3-ylidene acetonitrile (123 mg, 0.66 mmol), DBU (126 mg, 0.82 mmol), m. Compound E 0.3 g, yield 82.6%.
4)2-(3-(3-(7H-吡咯[2,3-d]嘧啶-4-基)-4,5,6,7-四氢-1H-吡唑[4,3-c]吡啶-1-基)-1-(乙基磺酰基)氮杂环丁烷-3-基)乙腈的制备 4) 2-(3-(3-(7H-pyrrole[2,3-d]pyrimidin-4-yl)-4,5,6,7-tetrahydro-1H-pyrazole [4,3-c] Preparation of pyridin-1-yl)-1-(ethylsulfonyl)azetidin-3-yl)acetonitrile
将叔丁基-1-(3-(腈甲基)-1-(乙基磺酰基)氮杂环丁烷-3-基)-3-(7-((2-(三甲基硅基)乙氧基)甲基)-7H-吡咯[2,3-d]嘧啶-4-基)-1,4,6,7-四氢-5H-吡唑[4,3-c]吡啶-5-甲酸酯(化合物E，0.30g，0.51mmol)加到三氟醋酸(10mL)中，氮气氛室温反应1小时。反应结束后，减压浓缩除去溶剂。加入氨水/甲醇/二氯甲烷(1:1:8)(20mL)，继续室温反应2小时。反应液经浓缩柱层析得目标化合物11 56mg，收率22.4％。tert-Butyl-1-(3-(nitrilemethyl)-1-(ethylsulfonyl)azetidin-3-yl)-3-(7-((2-(trimethylsilyl)) Ethoxy)methyl)-7H-pyrrole[2,3-d]pyrimidin-4-yl)-1,4,6,7-tetrahydro-5H-pyrazole[4,3-c]pyridine- 5-formate (Compound E, 0.30 g, 0.51 mmol) was added to trifluoroacetic acid (10 mL). After completion of the reaction, the solvent was concentrated under reduced pressure. Ammonia/methanol/methylene chloride (1:1:8) (20 mL) was added and the reaction was continued at room temperature for 2 hours. The reaction solution was subjected to concentration column chromatography to give the object compound 11 56 mg (yield: 22.4%).
1H-NMR(400MHz,DMSO-d6)δ(ppm)12.08(s,1H),8.76(s,1H),7.55(s,1H),7.11(s,1H),4.65-4.63(d,J＝8.16Hz,2H),4.25-4.23(d,J＝8.08Hz,2H),3.54(s,2H),3.25-3.23(d,J＝7.2Hz,2H),2.96(s,2H),2.69(s,2H),1.26-1.23(t,3H)；LC-MS(ESI+APCI)[M+H]+427.21。 1 H-NMR (400MHz, DMSO -d6) δ (ppm) 12.08 (s, 1H), 8.76 (s, 1H), 7.55 (s, 1H), 7.11 (s, 1H), 4.65-4.63 (d, J = 8.16 Hz, 2H), 4.25 - 4.23 (d, J = 8.08 Hz, 2H), 3.54 (s, 2H), 3.25-3.23 (d, J = 7.2 Hz, 2H), 2.96 (s, 2H), 2.69 (s, 2H), 1.26-1.23 (t, 3H); LC-MS (ESI+APCI) [M+H] + 427.21.
实施例12：2-(3-(3-(7H-吡咯[2,3-d]嘧啶-4-基)-4,5,6,7-四氢-1H-吲哚-1H-基)-1-(乙基磺酰基)氮杂环丁烷-3-基)乙腈(化合物12)的制备Example 12: 2-(3-(3-(7H-pyrrole[2,3-d]pyrimidin-4-yl)-4,5,6,7-tetrahydro-1H-indole-1H-yl) Preparation of 1-(ethylsulfonyl)azetidin-3-yl)acetonitrile (Compound 12)
Figure PCTCN2015090497-appb-000022
Figure PCTCN2015090497-appb-000022
合成路线synthetic route
Figure PCTCN2015090497-appb-000023
Figure PCTCN2015090497-appb-000023
1)2-(7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H--吡咯并[2,3-d]嘧啶-4-羰酰基)环己基-1-酮(化合物C)的制备1) 2-(7-((2-(Trimethylsilyl)ethoxy)methyl)-7H--pyrrolo[2,3-d]pyrimidin-4-carbonyl)cyclohexyl-1 - Preparation of ketone (Compound C)
氮气氛、0℃下将二异丙胺滴到丁基锂(160mL,0.4moL,10eq)中，搅拌1h，降温至-78℃，将环己酮(40.0g,0.4moL,10eq)滴入，保温搅拌2h。在-78℃下将化合物A(12.0g,0.04moL,1.0eq)的THF溶液(100mL)中滴入，自然升至室温，搅拌15h，TLC检测反应完全。反应液中加入饱和氯化铵溶液(50mL)，EA萃取 (100mL*3)，合并有机相旋干直接投下一步。Diisopropylamine was added dropwise to butyllithium (160 mL, 0.4 mol, 10 eq) at 0 ° C, stirred for 1 h, cooled to -78 ° C, and cyclohexanone (40.0 g, 0.4 mol, 10 eq) was added dropwise. Stir for 2 hours with heat. A solution of Compound A (12.0 g, 0.04 mol, 1.0 eq) in THF (100 mL) was added dropwise, and the mixture was warmed to room temperature and stirred for 15 h. Add saturated ammonium chloride solution (50 mL) to the reaction solution, extract with EA (100 mL * 3), combined organic phase spin directly into the next step.
2)4-(4,5,6,7-四氢-1H-吲唑-3-基)-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-吡咯并[2,3-d]嘧啶(化合物D)的制备2) 4-(4,5,6,7-Tetrahydro-1H-indazol-3-yl)-7-((2-(trimethylsilyl)ethoxy)methyl)-7H- Preparation of pyrrolo[2,3-d]pyrimidine (Compound D)
将上述化合物C溶于异丙醇(50mL)中，加入水合肼(20mL)，室温搅拌24h，TLC检测反应完全。反应液倒入水(50mL)中，EA(100mL*3)萃取，合并有机相，柱层析纯化得1.6g油状物。两步收率：10.7％。The above-mentioned compound C was dissolved in isopropyl alcohol (50 mL), hydrazine hydrate (20 mL) was added, and the mixture was stirred at room temperature for 24 hr. The reaction mixture was poured into water (50 mL), EtOAc (EtOAc m. Two-step yield: 10.7%.
3)2-(1-(乙基磺酰基)-3-(7-((2-(三甲基硅基)乙氧基)甲基)-7H-吡咯[2,3-d]嘧啶-4-基)-4,5,6,7-四氢-1H-吲唑-1-基)-氮杂环丁烷-3-基)乙腈(化合物E)的制备3) 2-(1-(ethylsulfonyl)-3-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrole[2,3-d]pyrimidine- Preparation of 4-yl)-4,5,6,7-tetrahydro-1H-indazol-1-yl)-azetidin-3-yl)acetonitrile (Compound E)
将4-(4,5,6,7-四氢-1H-吲唑-3-基)-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-吡咯并[2,3-d]嘧啶(化合物D，700mg,1.89mmoL,1.0eq)溶于乙腈(10mL)中，加入2-(1-(乙基磺酰基)氮杂环丁烷-3-亚基)乙腈(423mg,2.27mmoL,1.2eq)和DBU(275mg,2.27mmoL,1.2eq)，常温搅拌24h。反应液倒入水(5mL)中，EA(10mL*2)萃取，合并有机相，柱层析纯化得500mg白色固体。收率：47.49％。4-(4,5,6,7-Tetrahydro-1H-indazol-3-yl)-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrole And [2,3-d]pyrimidine (Compound D, 700 mg, 1.89 mmol, 1.0 eq) was dissolved in acetonitrile (10 mL) and 2-(1-(ethylsulfonyl)azetidin-3- Acetonitrile (423 mg, 2.27 mmol, 1.2 eq) and DBU (275 mg, 2.27 mmol, 1.2 eq), stirred at ambient temperature for 24 h. The reaction mixture was poured into water (5 mL), EtOAc (EtOAc) Yield: 47.49%.
4)2-(3-(3-(7H-吡咯[2,3-d]嘧啶-4-基)-4,5,6,7-四氢-1H-吲哚-1H-基)-1-(乙基磺酰基)氮杂环丁烷-3-基)乙腈(化合物12)的制备4) 2-(3-(3-(7H-pyrrole[2,3-d]pyrimidin-4-yl)-4,5,6,7-tetrahydro-1H-indole-1H-yl)-1 -(Ethylsulfonyl)azetidin-3-yl)acetonitrile (Compound 12)
2-(1-(乙基磺酰基)-3-(7-((2-(三甲基硅基)乙氧基)甲基)-7H-吡咯[2,3-d]嘧啶-4-基)-4,5,6,7-四氢-1H-吲唑-1-基)-氮杂环丁烷-3-基)乙腈(化合物E，500mg,0.89mmoL,1.0eq)溶于TFA(10mL)中，常温搅拌2h，TLC检测原料反应完。将反应液旋干，加入10％的氨水溶液，常温搅拌15h，有固体析出，过滤，滤饼用DCM(10mL)重结晶得129mg白色固体。收率：33.70％。2-(1-(ethylsulfonyl)-3-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrole[2,3-d]pyrimidine-4- -4,5,6,7-tetrahydro-1H-indazol-1-yl)-azetidin-3-yl)acetonitrile (Compound E, 500 mg, 0.89 mmol, 1.0 eq) was dissolved in TFA (10 mL), stirred at room temperature for 2 h, and the reaction of the starting material by TLC was completed. The reaction mixture was dried with EtOAc EtOAc EtOAc. Yield: 33.70%.
1H-NMR(400MHz,DMSO-d6)12.05(s,1H),8.76(s,1H),7.54(t,J＝2.96Hz,1H),7.09(qJ＝1.76Hz,1H),4.65(d,J＝8.84Hz,2H),4.23(d,J＝8.92Hz,2H),3.55(s,2H),3.24(q,J＝7.32Hz,2H),2.97(t,J＝5.64Hz,4H),2.70(t,J＝5.52Hz,2H),1.87-1.70(m,4H),1.25(t,J＝7.28Hz,3H)。LC-MS(ESI+APCI)[M+H]+426.10。 1 H-NMR (400MHz, DMSO -d6) 12.05 (s, 1H), 8.76 (s, 1H), 7.54 (t, J = 2.96Hz, 1H), 7.09 (qJ = 1.76Hz, 1H), 4.65 (d , J = 8.84 Hz, 2H), 4.23 (d, J = 8.92 Hz, 2H), 3.55 (s, 2H), 3.24 (q, J = 7.32 Hz, 2H), 2.97 (t, J = 5.64 Hz, 4H ), 2.70 (t, J = 5.52 Hz, 2H), 1.87-1.70 (m, 4H), 1.25 (t, J = 7.28 Hz, 3H). LC-MS (ESI+APCI) [M+H] + 426.
实施例13：2-(3-(3-(7H-吡咯[2,3-d]嘧啶-4-基)-5,6-二氢环戊基[c]吡唑-1(4H)-基)-1-(乙基磺酰基)氮杂环丁烷-3-基)乙腈(化合物13)的制备Example 13: 2-(3-(3-(7H-pyrrole[2,3-d]pyrimidin-4-yl)-5,6-dihydrocyclopentyl[c]pyrazole-1(4H)- Of ethyl-1-(ethylsulfonyl)azetidin-3-yl)acetonitrile (compound 13)
Figure PCTCN2015090497-appb-000024
Figure PCTCN2015090497-appb-000024
参照实施例12的合成方法，制备了化合物13，白色固体41mg，收率26.98％。 Referring to the synthesis method of Example 12, Compound 13 was obtained as a white solid (41 mg, yield: 26.98%).
1H-NMR(400MHz,DMSO-d6)12.08(s,1H),8.76(s,1H),7.56(t,J＝3.00Hz，1H),7.13(q,J＝1.8Hz 3.24Hz,1H),4.54(d,J＝8.96Hz,2H),4.23(d,J＝8.96Hz,2H),3.64(s,2H),3.25(q,J＝7.36Hz,2H),3.00-2.85(m,4H),2.66-2.55(m,2H),1.25(t,J＝7.28Hz,3H)。[M+H]+412.20。 1 H-NMR (400MHz, DMSO -d6) 12.08 (s, 1H), 8.76 (s, 1H), 7.56 (t, J = 3.00Hz, 1H), 7.13 (q, J = 1.8Hz 3.24Hz, 1H) , 4.54 (d, J = 8.96 Hz, 2H), 4.23 (d, J = 8.96 Hz, 2H), 3.64 (s, 2H), 3.25 (q, J = 7.36 Hz, 2H), 3.00-2.85 (m, 4H), 2.66-2.55 (m, 2H), 1.25 (t, J = 7.28 Hz, 3H). [M+H] + 412.20.
实施例14：3-(3-(7H-吡咯[2,3-d]嘧啶-4-基)-4,5,6,7-四氢-1H-吲哚-1H-基)-3-环戊基丙氰(化合物14)的制备Example 14: 3-(3-(7H-pyrrole[2,3-d]pyrimidin-4-yl)-4,5,6,7-tetrahydro-1H-indole-1H-yl)-3- Preparation of cyclopentylpropane cyanide (compound 14)
Figure PCTCN2015090497-appb-000025
Figure PCTCN2015090497-appb-000025
参照实施例12的合成方法，制备了化合物14，白色固体25mg，收率：31％。Referring to the synthesis method of Example 12, Compound 14 was obtained as a white solid, 25 mg, yield: 31%.
1H-NMR(400M Hz,DMSO-d6)11.98(s,1H),8.74(s,1H),7.51(t,J＝2.88Hz,1H),7.19(q,J＝1.84Hz,1H),4.41(m,1H),3.64(s,2H),2.67-3.25(m,7H),1.18-1.86(m,12H)。HPLC:97.0％,[M+H]+360.99。1H-NMR (400 M Hz, DMSO-d6) 11.98 (s, 1H), 8.74 (s, 1H), 7.51 (t, J = 2.88 Hz, 1H), 7.19 (q, J = 1.84 Hz, 1H), 4.41 (m, 1H), 3.64 (s, 2H), 2.67-3.25 (m, 7H), 1.18-1.86 (m, 12H). HPLC: 97.0%, [M+H] + 360.99.
实施例15：2-(3-(3-(7H-吡咯[2,3-d]嘧啶-4-基)-1H-吡咯[2,3-c]吡啶-1-基)-1-(1-(3-氟-2-(三氟甲基)异烟酰基)哌啶-4-基)氮杂环丁烷-3-基)乙腈(化合物15)的制备Example 15: 2-(3-(3-(7H-pyrrole[2,3-d]pyrimidin-4-yl)-1H-pyrrole[2,3-c]pyridin-1-yl)-1-( Preparation of 1-(3-fluoro-2-(trifluoromethyl)isonicotinylpiperidin-4-yl)azetidin-3-yl)acetonitrile (Compound 15)
Figure PCTCN2015090497-appb-000026
Figure PCTCN2015090497-appb-000026
合成路线：synthetic route:
Figure PCTCN2015090497-appb-000027
Figure PCTCN2015090497-appb-000027
1)叔丁基-3-(氰基甲基)-3-(3-(7-((2-(三甲基硅基)乙氧基)甲基)-7H-吡咯[2,3-d]嘧啶-4-基)-1H-吡咯[2,3-c]吡啶-1-基)氮杂环丁烷-1-酰胺(化合物C)的制备 1) tert-Butyl-3-(cyanomethyl)-3-(3-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrole [2,3- Preparation of d]pyrimidin-4-yl)-1H-pyrrole[2,3-c]pyridin-1-yl)azetidin-1-amide (Compound C)
将4-(1H-吡咯[2,3-c]吡啶-3-基)-7-((2-(三甲基硅基)乙氧基)甲基)-7H-吡咯[2,3-d]嘧啶(0.360g,1.0mmol)、3-(氰基亚甲基)氮杂环丁烷-1-甲酸叔丁酯(0.240g,1.2mmol)、5mL乙腈和DBU(0.18ml,1.2mmol)加到50mL茄形瓶中，氮气保护，室温搅拌36小时。反应结束后减压浓缩制砂过柱(PE:EA＝5:1)得目标化合物0.29g，收率53.32％。4-(1H-pyrrole[2,3-c]pyridin-3-yl)-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrole [2,3- d]pyrimidine (0.360 g, 1.0 mmol), 3-(cyanomethylene)azetidin-1-carboxylic acid tert-butyl ester (0.240 g, 1.2 mmol), 5 mL acetonitrile and DBU (0.18 ml, 1.2 mmol ) was added to a 50 mL eggplant-shaped bottle, protected with nitrogen, and stirred at room temperature for 36 hours. After the completion of the reaction, the residue was filtered under reduced pressure (PE: EA = 5:1) to afford 0.29 g of the desired compound.
2)2-(3-(3-(7-((2-(三甲基硅基)乙氧基)甲基)-7H-吡咯[2,3-d]嘧啶-4-基)-1H-吡咯[2,3-c]吡啶-1-基)氮杂环丁烷-3-基)乙腈(化合物D)的制备2) 2-(3-(3-(7-((2-(Trimethylsilyl)ethoxy)methyl)-7H-pyrrole[2,3-d]pyrimidin-4-yl)-1H -Preparation of pyrrole [2,3-c]pyridin-1-yl)azetidin-3-yl)acetonitrile (Compound D)
叔丁基-3-(氰基甲基)-3-(3-(7-((2-(三甲基硅基)乙氧基)甲基)-7H-吡咯[2,3-d]嘧啶-4-基)-1H-吡咯[2,3-c]吡啶-1-基)氮杂环丁烷-1-酰胺(化合物C，0.12g，0.22mmol)溶解于4ml DCM中，降温0℃滴加TFA(0.08ml，1.09mmol)，反应16小时，旋干直接投下一步。tert-Butyl-3-(cyanomethyl)-3-(3-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrole [2,3-d] Pyrimidin-4-yl)-1H-pyrrole[2,3-c]pyridin-1-yl)azetidin-1-amide (Compound C, 0.12 g, 0.22 mmol) was dissolved in 4 mL DCM. TFA (0.08 ml, 1.09 mmol) was added dropwise at °C, and the mixture was reacted for 16 hr.
3)2-(1-(1-(3-氟-2-(三氟甲基)异烟酰基)哌啶-4-基)-3-(3-(7-((2-(三甲基硅基)乙氧基)甲基-7H-吡咯[2,3-d]嘧啶-4-基)-1H-吡咯[2,3-c]吡啶-1-基)氮杂环丁烷-3-基)乙腈(化合物F)的制备3) 2-(1-(1-(3-Fluoro-2-(trifluoromethyl)isonicotinyl)piperidin-4-yl)-3-(3-(7-((2-()) Ethyl) ethoxy)methyl-7H-pyrrole[2,3-d]pyrimidin-4-yl)-1H-pyrrole[2,3-c]pyridin-1-yl)azetidine- Preparation of 3-yl)acetonitrile (Compound F)
2-(3-(3-(7-((2-(三甲基硅基)乙氧基)甲基)-7H-吡咯[2,3-d]嘧啶-4-基)-1H-吡咯[2,3-c]吡啶-1-基)氮杂环丁烷-3-基)乙腈(化合物D，460mg)，1-(3-氟-2-(三氟甲基)异烟酰基)哌啶-4-酮(化合物E，290mg)加到无水甲醇4mL中，加入氰基硼氢化钠60mg，室温搅拌3小时。反应结束加入饱和碳酸氢钠水溶液5ml，EA 20mL*3萃取，合并有机相，饱和食盐水5ml洗一遍，硫酸钠干燥，制砂过柱，PE:EA＝2:1-1:2得目标化合物F 100mg，用于下步反应。2-(3-(3-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrole[2,3-d]pyrimidin-4-yl)-1H-pyrrole [2,3-c]pyridin-1-yl)azetidin-3-yl)acetonitrile (Compound D, 460 mg), 1-(3-fluoro-2-(trifluoromethyl)isonicotinyl) Piperidin-4-one (Compound E, 290 mg) was added to 4 mL of anhydrous methanol, and 60 mg of sodium cyanoborohydride was added, and the mixture was stirred at room temperature for 3 hours. At the end of the reaction, 5 ml of a saturated aqueous solution of sodium hydrogencarbonate and EA 20 mL*3 were added, and the organic phase was combined, washed with 5 ml of saturated brine, dried over sodium sulfate, and then passed to a column, PE:EA=2:1:1:2 to obtain the target compound F 100 mg for the next step of the reaction.
4)2-(3-(3-(7H-吡咯[2,3-d]嘧啶-4-基)-1H-吡咯[2,3-c]吡啶-1-基)-1-(1-(3-氟-2-(三氟甲基)异烟酰基)哌啶-4-基)氮杂环丁烷-3-基)乙腈(化合物15)的制备4) 2-(3-(3-(7H-pyrrole[2,3-d]pyrimidin-4-yl)-1H-pyrrole[2,3-c]pyridin-1-yl)-1-(1- Preparation of (3-fluoro-2-(trifluoromethyl)isonicotinyl)piperidin-4-yl)azetidin-3-yl)acetonitrile (Compound 15)
2-(1-(1-(3-氟-2-(三氟甲基)异烟酰基)哌啶-4-基)-3-(3-(7-((2-(三甲基硅基)乙氧基)甲基-7H-吡咯[2,3-d]嘧啶-4-基)-1H-吡咯[2,3-c]吡啶-1-基)氮杂环丁烷-3-基)乙腈(0.100g，0.14mmol)溶解于2.0mlTFA中，22℃搅拌40min，TLC显示原料消失，抽干溶剂，加入3ml10％氨水甲醇溶液，22℃搅拌12小时，抽干溶剂，制砂过柱，PE:EA＝1:1，得0.070g白色固体，收率85.02％。2-(1-(1-(3-fluoro-2-(trifluoromethyl)isonicotinyl)piperidin-4-yl)-3-(3-(7-((2-(trimethylsilane)) Ethyl)ethyl-7H-pyrrole[2,3-d]pyrimidin-4-yl)-1H-pyrrole[2,3-c]pyridin-1-yl)azetidin-3- Acetonitrile (0.100 g, 0.14 mmol) was dissolved in 2.0 ml of TFA, stirred at 22 ° C for 40 min, TLC showed the disappearance of the starting material, solvent was evaporated, 3 ml of 10% aqueous ammonia solution was added, and the mixture was stirred at 22 ° C for 12 hours, and the solvent was drained. Column, PE: EA = 1:1, yielding 0.070 g of a white solid, yield 85.02%.
LC-MS：Found 603.9。1H-NMR(400MHz，DMSO-d6)，δ(ppm):12.14(s，1H)，8.80-8.82(m,2H)，8.60-8.68(m,2H)，8.37-8.39(m,2H)，8.16(s，1H)，7.91(t，1H)，7.62(s，1H)，7.08(d，1H)，4.04-4.08(m，3H)，3.71-3.75(m，4H)，3.48(s，1H)，3.08(m,1H),1.66-1.79(m,2H),1.29-1.38(m,2H)。</ RTI> <RTIgt; 8.39 (m, 2H), 8.16 (s, 1H), 7.91 (t, 1H), 7.62 (s, 1H), 7.08 (d, 1H), 4.04-4.08 (m, 3H), 3.71-3.75 (m, 4H), 3.48 (s, 1H), 3.08 (m, 1H), 1.66-1.79 (m, 2H), 1.29-1.38 (m, 2H).
实施例16：2-(3-(3-(7H-吡咯[2,3-d]嘧啶-4-基)-1H-吡咯[2,3-b]吡啶-1-基)-1-(1-(3-氟-2-(三氟甲基)异烟酰基)哌啶-4-基)氮杂环丁烷-3-基)乙腈(化合物16)的制备 Example 16: 2-(3-(3-(7H-pyrrole[2,3-d]pyrimidin-4-yl)-1H-pyrrole[2,3-b]pyridin-1-yl)-1-( Preparation of 1-(3-fluoro-2-(trifluoromethyl)isonicotinylpiperidin-4-yl)azetidin-3-yl)acetonitrile (Compound 16)
Figure PCTCN2015090497-appb-000028
Figure PCTCN2015090497-appb-000028
参考实施例15的合成方法，制备了化合物16，白色固体125mg，收率：41％。Referring to the synthesis method of Example 15, Compound 16 was obtained as a white solid, 125 mg, yield: 41%.
HPLC：96.99％，LC-MS：Found 603.9。1H-NMR(400MHz，DMSO-d6)，δ(ppm):12.12(s,1H)，9.02(d,1H),8.81(s,1H),8.67(d,1H),8.37-8.40(m,2H),7.91(t,1H),7.62(s,1H),7.34(t,1H),7.10(s,1H),4.06(m，1H),3.89-3.96(m，2H)，3.78-3.79(m，2H)，3.61(s，2H)，3.47(s，1H)，3.08(m,1H),1.66-1.79(m,2H),1.31-1.38(m,2H)。HPLC: 96.99%, LC-MS: Found 603.9. 1 H-NMR (400 MHz, DMSO-d6), δ (ppm): 12.12 (s, 1H), 9.02 (d, 1H), 8.81 (s, 1H), 8.67 (d, 1H), 8.37-8.40 (m) , 2H), 7.91 (t, 1H), 7.62 (s, 1H), 7.34 (t, 1H), 7.10 (s, 1H), 4.06 (m, 1H), 3.89-3.96 (m, 2H), 3.78- 3.79 (m, 2H), 3.61 (s, 2H), 3.47 (s, 1H), 3.08 (m, 1H), 1.66-1.79 (m, 2H), 1.31-1.38 (m, 2H).
实施例17：Example 17
采用体Caliper Mobility Shift Assay方法在不同三磷酸腺苷(ATP)浓度下Km值研究检测本发明下述列举化合物对Janus系列激酶的抑制剂的作用。发现本发明化合物对JAK-STAT途径的抑制作用非常明显。部分数据如下：The Km value of the different adenosine triphosphate (ATP) concentrations was examined by the Caliper Mobility Shift Assay method to examine the effects of the following enumerated compounds of the invention on inhibitors of the Janus series of kinases. The inhibitory effect of the compounds of the invention on the JAK-STAT pathway was found to be very pronounced. Some of the data is as follows:
Figure PCTCN2015090497-appb-000029
Figure PCTCN2015090497-appb-000029
实施例18Example 18
利用MTS法，检测化合物对IL-3、IL-4诱导的TF-1细胞增殖的抑制情况，并计算IC50值。The inhibition of IL-3 and IL-4 induced proliferation of TF-1 cells by the compounds was examined by the MTS method, and the IC 50 value was calculated.
TF-1细胞于无血清培养基中饥饿过夜后，分别用含10ng/ml IL-3、IL-4 RPMI-1640培养基(含10％FBS)重悬，以15000cell/well密度接入96孔板中。实验设两个对照组。阴性对照组：含细胞，但不含有IL-3或IL-4因子，含有10％血清；阳性对照组(化合物0浓度孔)：无化合物作用的细胞，但含有IL-3或IL-4因 子以及血清。向细胞中加入梯度稀释的化合物，使终浓度为20000nM起，3倍稀释，一共10个浓度(含0浓度孔)，每浓度3复孔。培养基中DMSO含量为0.1％。将细胞置于37℃，5％CO2的培养箱中培养，使化合物作用于细胞72h。加入CCK8，每孔含10％(V/V)，37℃孵育3h。酶标仪检测450nM处吸光值。After starving overnight in serum-free medium, TF-1 cells were resuspended in 10 ng/ml IL-3, IL-4 RPMI-1640 medium (containing 10% FBS), and inserted into 96 wells at a density of 15000 cells/well. In the board. Two control groups were set up in the experiment. Negative control group: containing cells, but not containing IL-3 or IL-4 factor, containing 10% serum; positive control group (compound 0 concentration well): cells without compound action, but containing IL-3 or IL-4 Child and serum. Gradiently diluted compounds were added to the cells to a final concentration of 20,000 nM, diluted 3 times, a total of 10 concentrations (containing 0 concentration wells), 3 replicates per concentration. The DMSO content in the medium was 0.1%. The cells were cultured in an incubator at 37 ° C, 5% CO 2 , and the compounds were allowed to act on the cells for 72 h. CCK8 was added, containing 10% (V/V) per well, and incubated at 37 °C for 3 h. The microplate reader detects the absorbance at 450 nM.
按以下公式计算抑制率：Calculate the inhibition rate according to the following formula:
Figure PCTCN2015090497-appb-000030
Figure PCTCN2015090497-appb-000030
IC50值的计算：以log[化合物浓度]为横坐标，Inhibition％为纵坐标，在GraphPad Prism 5中，拟合非线性曲线：log(inhibitor)vs.response--Variable slope，并计算IC50值。Calculation of IC 50 value: log [compound concentration] as the abscissa, Inhibition% as the ordinate, and in GraphPad Prism 5, fit the nonlinear curve: log(inhibitor) vs.response--Variable slope, and calculate IC 50 value.
Figure PCTCN2015090497-appb-000031
Figure PCTCN2015090497-appb-000031
实施例19Example 19
本发明化合物抑制类风湿性关节炎的作用，选择DBA/1J小鼠，将50ug牛Ⅱ型胶原与等体积完全弗氏佐 剂(CFA)完全乳化后皮下注射。21天后以50ug相同抗原与不完全弗氏佐剂(IFA充分乳化后，加强免疫1次。从第45天开始观察记录。采用1-4计分法：1分，正常；2分，1个关节肿胀；3分，超过1个关节肿胀，但并未累积全部关节；4分，整个爪的严重肿胀或强直。每只爪的评分相加即得到小鼠关节炎症的总评分。关节总评分大于1的小鼠为模型建立成功。成功建立小鼠类风湿关节炎模型后采用本发明化合物给小鼠灌胃给药，给药2周后对小鼠的关节炎症进行评分，结果显示本品对小鼠类风湿性关节炎有明显的治疗作用。部分数据如下：The compound of the present invention inhibits the action of rheumatoid arthritis, selects DBA/1J mice, and 50 ug of bovine type II collagen with an equal volume of complete Freund's The agent (CFA) was completely emulsified and injected subcutaneously. After 21 days, 50 ug of the same antigen and incomplete Freund's adjuvant (IFA fully emulsified, boosted once. The observation was started from the 45th day. Using 1-4 scoring method: 1 point, normal; 2 points, 1 Joint swelling; 3 points, more than 1 joint swelling, but did not accumulate all joints; 4 points, the entire paw was severely swollen or stiff. The score of each claw was added to obtain the total score of joint inflammation in mice. Mice greater than 1 were successfully established in the model. After successfully establishing a mouse model of rheumatoid arthritis, the mice of the present invention were intragastrically administered with the compound of the present invention, and the joint inflammation of the mice was scored after 2 weeks of administration, and the result showed that the product was displayed. It has obvious therapeutic effects on rheumatoid arthritis in mice. Some data are as follows:
化合物组别Compound group 关节炎评分Arthritis score
ControlControl 11
生理盐水对照组Saline control group 44
实施例1化合物Example 1 compound 1.91.9
实施例2化合物Example 2 compound 2.02.0
实施例3化合物Example 3 compound 1.81.8
实施例4化合物Example 4 compound 1.61.6
实施例5化合物Example 5 compound 2.12.1
实施例6化合物Example 6 compound 2.32.3
JAK抑制剂BaricitinibJAK inhibitor Baricitinib 2.02.0

Claims (9)

  1. 如化学结构式(I)所示的化合物或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、或其混合物形式、或药学上可接受的盐：a compound of the formula (I), or a tautomer, a mesophil, a racemate, an enantiomer, a diastereomer, or a mixture thereof, or a pharmaceutically acceptable compound Accepted salt:
    Figure PCTCN2015090497-appb-100001
    Figure PCTCN2015090497-appb-100001
    其中，R1选自氢、卤素、烷基；Wherein R 1 is selected from the group consisting of hydrogen, halogen, and alkyl;
    R2选自：R 2 is selected from:
    Figure PCTCN2015090497-appb-100002
    Figure PCTCN2015090497-appb-100003
    其中R4、R5、R6、R7各自独立的选自氢、卤素、烷基、硝基、氨基、羟基；X选自O、N或S；
    Figure PCTCN2015090497-appb-100002
    Figure PCTCN2015090497-appb-100003
    Wherein R 4 , R 5 , R 6 , and R 7 are each independently selected from the group consisting of hydrogen, halogen, alkyl, nitro, amino, and hydroxy; and X is selected from O, N or S;
    R3选自：R 3 is selected from:
    Figure PCTCN2015090497-appb-100004
    Figure PCTCN2015090497-appb-100005
    Figure PCTCN2015090497-appb-100004
    Figure PCTCN2015090497-appb-100005
    其中R8选自：烷基，环烷基，杂环烷基。Wherein R 8 is selected from the group consisting of alkyl, cycloalkyl and heterocycloalkyl.
  2. 根据权利要求1所述的如化学结构式(I)所示的化合物或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、或其混合物形式、或药学上可接受的盐，其特征在于，R2中R4、R5、R6、R7各自独立的选自氢、卤素、烷基。 A compound of the formula (I), or a tautomer, a mesomer, a racemate, an enantiomer, a diastereomer thereof, or a compound thereof according to claim 1 a mixture, or a pharmaceutically acceptable salt thereof, wherein, R 2 in R 4, R 5, R 6 , R 7 are each independently selected from hydrogen, halogen, alkyl.
  3. 根据权利要求1所述，R3选自
    Figure PCTCN2015090497-appb-100006
    其中R8选自：烷基，环烷基，杂环烷基。    According to claim 1, R 3 is selected from
    Figure PCTCN2015090497-appb-100006
    Wherein R 8 is selected from the group consisting of alkyl, cycloalkyl and heterocycloalkyl.
  4. 根据权利要求1-3所述的如化学结构式(I)所示的化合物或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、或其混合物形式、或药学上可接受的盐，其特征在于其选自下述化合物中任一：a compound of the formula (I), or a tautomer, a mesophil, a racemate, an enantiomer, a diastereomer thereof, according to claims 1-3, Or a mixture thereof, or a pharmaceutically acceptable salt, characterized in that it is selected from any of the following compounds:
    Figure PCTCN2015090497-appb-100007
    Figure PCTCN2015090497-appb-100007
  5. 一种药用组合物，其包含治疗有效量的权利要求1-4中任意一项的化合物、其立体异构体、互变异构体或药学上可接受的盐，以及药学上可接受的载体、辅助剂或溶媒。A pharmaceutical composition comprising a therapeutically effective amount of a compound according to any one of claims 1 to 4, a stereoisomer, tautomer or pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable Carrier, adjuvant or vehicle.
  6. 权利要求1-4中任意一项的化合物、其立体异构体、互变异构体、前药或药学上可接受的盐，其用作药物。 A compound, a stereoisomer, tautomer, prodrug or pharmaceutically acceptable salt thereof, according to any one of claims 1 to 4, for use as a medicament.
  7. 权利要求1-4中任意一项的化合物、其立体异构体、互变异构体、前药或药学上可接受的盐，其用作制备治疗免疫疾病的药物。A compound, a stereoisomer, tautomer, prodrug or pharmaceutically acceptable salt thereof according to any one of claims 1 to 4 for use as a medicament for the preparation of a medicament for the treatment of an immune disease.
  8. 权利要求1-4中任意一项的化合物、其立体异构体、互变异构体、前药或药学上可接受的盐，其用作制备治疗选自类风湿性关节炎、哮喘、***性红斑狼疮、银屑病、IBD和移植排斥的免疫疾病的药物。A compound, a stereoisomer, a tautomer, a prodrug or a pharmaceutically acceptable salt thereof, according to any one of claims 1 to 4, which is used for the preparation of a treatment selected from the group consisting of rheumatoid arthritis, asthma, system Drugs for immune diseases of lupus erythematosus, psoriasis, IBD, and transplant rejection.
  9. 权利要求1-4中任意一项的化合物、其立体异构体、互变异构体、前药或药学上可接受的盐在制备治疗对JAK1激酶活性的抑制有反应的疾病的药物中的用途，所述疾病特别是免疫疾病，更特别是选自类风湿性关节炎、哮喘、***性红斑狼疮、银屑病、IBD和移植排斥的免疫疾病。 The compound according to any one of claims 1 to 4, a stereoisomer, tautomer, prodrug or pharmaceutically acceptable salt thereof, in the preparation of a medicament for treating a disease responsive to inhibition of JAK1 kinase activity Use, the disease is in particular an immune disease, more particularly an immune disease selected from the group consisting of rheumatoid arthritis, asthma, systemic lupus erythematosus, psoriasis, IBD and transplant rejection.
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