WO2016082737A1 - 一种取代的噻吩并嘧啶化合物及其制备方法 - Google Patents

一种取代的噻吩并嘧啶化合物及其制备方法 Download PDF

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WO2016082737A1
WO2016082737A1 PCT/CN2015/095384 CN2015095384W WO2016082737A1 WO 2016082737 A1 WO2016082737 A1 WO 2016082737A1 CN 2015095384 W CN2015095384 W CN 2015095384W WO 2016082737 A1 WO2016082737 A1 WO 2016082737A1
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substituted
unsubstituted
compound
group
pharmaceutically acceptable
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French (fr)
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易崇勤
许恒
韩方斌
林松文
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北大方正集团有限公司
方正医药研究院有限公司
北大医疗产业集团有限公司
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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
    • C07D495/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having sulfur atoms as the only ring hetero atoms
    • C07D495/02—Heterocyclic compounds containing in the condensed system at least one hetero ring having sulfur atoms as the only ring hetero atoms in which the condensed system contains two hetero rings
    • C07D495/04—Ortho-condensed systems

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  • the invention relates to a thienopyrimidine compound, belonging to the technical field of medicinal chemistry.
  • Molecularly targeted drugs are a key mode of action for receptors or transduction processes in the process of cell carcinogenesis, and a therapeutic mode that inhibits tumor growth at the molecular level. Targeting the characteristic molecules of tumor cells, it exerts anti-tumor effects while reducing the toxic side effects on normal cells.
  • Phosphoinositide 3-kinase catalyzes the synthesis of phosphatidylinositol (PI) second messenger PI(3)P, PI(3,4)P2 and PI(3,4,5)P3(PIP3) . Under the proper cellular environment, these three lipids control a variety of physiological processes including cell growth, survival, differentiation, and chemotaxis.
  • the major PI3-kinase isoform in cancer is a class I PI3-K ⁇ consisting of a catalytic (p110 ⁇ ) subunit and a adaptor (p85) subunit.
  • PI3-K/Akt/PTEN pathway Several components of the PI3-K/Akt/PTEN pathway are involved in tumor formation and play an important role in the regulation of many cellular processes including cell cycle regulation, proliferation, survival, apoptosis, and exercise, and are such as cancer, diabetes, and An important component of the molecular mechanism of diseases such as immune inflammation.
  • PI3-K ⁇ is an attractive and promising target for the development of cancer drugs, which will inhibit the proliferation of cancer cells, and avoid damage to the body by cytotoxic agents, and the tolerance of cancer cells to cytotoxic agents.
  • PI3-K ⁇ inhibitors for use as cancer therapeutics in the field of cancer therapy.
  • the present invention provides substituted thienopyrimidine compounds, as well as processes for their preparation and their use as PI3-K and/or mTOR kinase inhibitors.
  • R 1 is selected from substituted or unsubstituted aryl, substituted or unsubstituted aromatic heterocyclic group, the hetero atom of the aromatic heterocyclic group is selected from N, S and O;
  • R 2 is selected from the group consisting of hydrogen and C1-C4 alkyl
  • R 3 is selected from the group consisting of C 1 -C 4 alkoxy, C 1 -C 4 alkyl, halogen (fluoro, chloro, bromo, iodo);
  • X is C or N
  • R 4 is selected from hydrogen, halogen, substituted or unsubstituted C1-C4 alkyl, substituted or unsubstituted C1-C4 alkoxy, substituted or unsubstituted amino, substituted or unsubstituted sulfonate. Acylamino.
  • R 1 is selected from a substituted or unsubstituted phenyl group, a substituted or unsubstituted pyridyl group, a substituted or unsubstituted pyranyl group, a substituted or unsubstituted pyridazinyl group, a substituted or unsubstituted pyrimidinyl group, and a substitution.
  • unsubstituted pyrazinyl substituted or unsubstituted piperazinyl, substituted or unsubstituted furyl, substituted or unsubstituted thienyl, substituted or unsubstituted pyrrolyl, substituted or unsubstituted pyrazolyl, Substituted or unsubstituted imidazolyl, substituted or unsubstituted oxazolyl, substituted or unsubstituted thiazolyl, substituted or unsubstituted isoxazolyl.
  • the number of the substituents may be 1 to 5, and the substituent is selected from the group consisting of halogen, cyano, trifluoromethyl, C1-C4 alkoxy, C1-C4 alkane. Base, amino and hydroxyl groups.
  • R 1 is pyridyl
  • the substituted thienopyrimidine compound is a compound of formula II:
  • the sulfonylamino group is -NH-SO 2 -R 5 ; wherein R 5 is selected from a substituted or unsubstituted C1-C4 alkyl group, a substituted or unsubstituted C3-C7 cycloalkyl group, a substituted or unsubstituted phenyl group, a substituted or unsubstituted five- or six-membered aromatic heterocyclic group, the hetero atom of the aromatic heterocyclic ring being selected from the group consisting of N, S and O .
  • R 5 is selected from a substituted or unsubstituted C1-C4 alkyl group, a substituted or unsubstituted C3-C7 cycloalkyl group, a substituted or unsubstituted phenyl group, a substituted or unsubstituted pyridyl group, a substituted or unsubstituted group.
  • Pyranyl substituted or unsubstituted pyridazinyl, substituted or unsubstituted pyrimidinyl, substituted or unsubstituted pyrazinyl, substituted or unsubstituted piperazinyl, substituted or unsubstituted furanyl, substituted or Unsubstituted thienyl, substituted or unsubstituted pyrrolyl, substituted or unsubstituted pyrazolyl, substituted or unsubstituted imidazolyl, substituted or unsubstituted oxazolyl, substituted or unsubstituted thiazolyl, substituted or Unsubstituted isoxazolyl.
  • R 5 group has a substituent
  • the number of the substituent is 1-5, and these substituents are selected from halogen (fluorine, chlorine, bromine, iodine), cyano, trifluoromethyl , C1-C4 alkoxy group, C1-C4 alkyl group, amino group, hydroxyl group.
  • R 5 may be selected from the following groups:
  • the compound of formula I can be selected from the following compounds 1-13:
  • the present invention also provides a process for the preparation of the compound of the formula I, which comprises: subjecting the compound of the formula A and the compound of the formula B to the substitution reaction in the presence of a palladium catalyst, thereby obtaining the formula I Compound:
  • R 1 , R 2 , R 3 , R 4 and X are as defined above; R 6 is
  • the palladium catalyst may be tetrakis(triphenylphosphine)palladium, bis(triphenylphosphine)palladium dichloride or [1,1'-bis(diphenylphosphino)ferrocene. ] Palladium dichloride.
  • the compound represented by the formula A can be produced by the following method:
  • R 1 and R 2 are as defined above; R 7 is
  • the palladium catalyst may be tetrakis(triphenylphosphine)palladium, bis(triphenylphosphine)palladium dichloride or [1,1'-bis(diphenylphosphino)ferrocene]dichloride Palladium.
  • the compound represented by the above formula G can be produced by the following method:
  • the compound of formula F is first treated with a solution of hydrogen chloride in diethyl ether and then reacted with sodium iodide to give the compound of formula G:
  • R2 is as defined above.
  • the compound represented by the above formula F can be produced by the following method:
  • the compound of the formula E is reacted with a chlorinating reagent such as phosphorus oxychloride, thionyl chloride or oxalyl chloride to give a compound of the formula F:
  • R2 is as defined above.
  • the compound of the formula D is reacted with a bromine reagent such as liquid bromine or N-bromosuccinimide to obtain a compound of the formula E:
  • R 2 is as defined above.
  • the compound represented by the above formula D can be produced by the following method:
  • the compound represented by the formula C is subjected to the following cyclization reaction with a reagent such as formamidine or formamide to obtain a compound represented by the formula D:
  • R 2 is as defined above.
  • the substituted thienopyrimidine compound of the present invention is a multi-kinase inhibitor capable of simultaneously inhibiting a variety of kinases present in cells and on the cell surface, including PI3-K and/or mTOR kinase.
  • the PI3-K priority refers to PI3-K ⁇ .
  • the compound and a pharmaceutically acceptable salt thereof are useful for the preparation of a medicament for treating and/or preventing a disease caused by PI3-K and/or mTOR kinase.
  • the pharmaceutically acceptable salts include: hydrochloride, hydrobromide, sulfate, phosphate, methanesulfonate, triflate, besylate, p-toluenesulfonate, 1- Naphthalene sulfonate, 2-naphthalene sulfonate, acetate, trifluoroacetate, malate, tartrate, citrate, lactate, oxalate, succinate, fumarate, Maleate, benzoate, salicylate, phenylacetate, mandelate, and the like.
  • the invention also provides a pharmaceutical composition for treating and/or preventing a disease caused by PI3-K and/or mTOR kinase, the pharmaceutical composition comprising the substituted thienopyrimidine compound of the invention and a medicinal adjuvant .
  • the pharmaceutical composition is an injection, an oral preparation, a transdermal absorbent or a suppository.
  • Diseases caused by PI3-K and/or mTOR kinase such as cancer, for example, brain cancer, glioblastoma, head and neck cancer, lung cancer, melanoma, liver cancer, kidney cancer, acute leukemia, chronic leukemia, non-small cell lung cancer, prostate Cancer, thyroid cancer, skin cancer, colon cancer, rectal cancer, pancreatic cancer, ovarian cancer, breast cancer, abnormal bone marrow hyperplasia syndrome, esophageal cancer, sarcoma, osteosarcoma or rhabdomyosarcoma.
  • cancer for example, brain cancer, glioblastoma, head and neck cancer, lung cancer, melanoma, liver cancer, kidney cancer, acute leukemia, chronic leukemia, non-small cell lung cancer, prostate Cancer, thyroid cancer, skin cancer, colon cancer, rectal cancer, pancreatic cancer, ovarian cancer, breast cancer, abnormal bone marrow hyperplasia syndrome, esophageal cancer, sarcoma,
  • Substituted thienopyrimidine compounds and pharmaceutically acceptable salts thereof according to the invention can be used to prepare drugs for treating and/or preventing tumor diseases.
  • 6-Bromo-4-(pyridin-4-yl)thieno[2,3-d]pyrimidine (Compound 1-6) (102 mg, 0.35 mmol), N-(2-methoxy-5-(4) ,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-3-yl)-2,4-difluorobenzenesulfonamide (179 mg, 0.42 mmol , 1.2 eq) and a mixture of 2N potassium carbonate (0.70 mL, 1.4 mmol, 4.0 eq) in dioxane (5 mL) were degassed, then bis(triphenylphosphine)palladium dichloride (26 mg, 0.035 mmol) , 0.1 eq).
  • reaction mixture was degassed and backfilled with argon (three cycles), and then stirred at 100 ° C for 6 hours under an argon atmosphere.
  • the reaction mixture was concentrated with EtOAc EtOAc m.
  • the combined organic layers were washed with EtOAcq.
  • the residue was purified by EtOAc EtOAcqqqqqqqq
  • Compound 2 was prepared in a similar manner to Compound 1 prepared in Example 1, except that N-(2-methoxy-5-(4,4,5,5-tetramethyl-1,3,2) was used in step 6. -dioxaborolan-2-yl)pyridin-3-yl)methanesulfonamide in place of N-(2-methoxy-5-(4,4,5,5-tetramethyl-1,3) , 2-dioxaborolan-2-yl)pyridin-3-yl)-2,4-difluorobenzenesulfonamide.
  • Compound 3 was prepared in a similar manner to Compound 1 prepared in Example 1, except that in step 6, 2-methoxypyridine-5-boronic acid was used instead of N-(2-methoxy-5-(4,4,5, 5-Tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-3-yl)-2,4-difluorobenzenesulfonamide.
  • Compound 4 was prepared in a similar manner to Compound 1 prepared in Example 1, except that in step 6, 2-methoxypyrimidine-5-boronic acid was used instead of N-(2-methoxy-5-(4,4,5, 5-Tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-3-yl)-2,4-difluorobenzenesulfonamide.
  • Compound 5 was prepared in a similar manner to Compound 1 prepared in Example 1, except that in step 6, 2-methoxy-3-fluoropyridine-5-boronic acid was used instead of N-(2-methoxy-5-(4, 4,5,5-Tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-3-yl)-2,4-difluorobenzenesulfonamide.
  • Compound 6-2 was prepared in a similar manner to the preparation of compound 1-2 in Step 1 in Example 1, except that 2-amino-4-methylthiophene-3-carboxylic acid methyl ester (6-1) was used instead of 2-amino. Methyl thiophene-3-carboxylate (1-1).
  • Compound 6-3 was prepared in a similar manner to the preparation of compound 1-3 in Step 2 of Example 1, except that 5-methylthieno[2,3-d]pyrimidin-4-ol (Compound 6-2) was used instead of thiophene. And [2,3-d]pyrimidin-4-phenol (Compound 1-2).
  • Compound 6-4 was prepared in a similar manner to the preparation of compound 1-4 in Step 3 of Example 1, except that 6-bromo-5-methylthieno[2,3-d]pyrimidin-4-ol (Compound 6- 3) Instead of 6-bromothieno[2,3-d]pyrimidin-4-ol (compounds 1-3).
  • Compound 6-5 was prepared in a similar manner to the preparation of compound 1-5 in Step 4 of Example 1, except that 6-bromo-4-chloro-5-methylthieno[2,3-d]pyrimidine (Compound 6- 4) Instead of 6-bromo-4-chlorothieno[2,3-d]pyrimidine (compounds 1-4).
  • Compound 6-6 was prepared in a similar manner to the preparation of compound 1-6 in Step 5 of Example 1, except that 6-bromo-4-iodo-5-methylthieno[2,3-d]pyrimidine (Compound 6- 5) Instead of 6-bromo-4-iodothieno[2,3-d]pyrimidine (compounds 1-5).
  • Step 6 N-(2-Methoxy-5-(5-methyl-4-(pyridin-4-yl)thieno[2,3-d]pyrimidin-6-yl)pyridin-3-yl) Synthesis of -2,4-difluorobenzenesulfonamide (Compound 6)
  • Compound 6 was prepared in a similar manner to the preparation of compound 1 in step 1 of Example 1, except that 6-bromo-5-methyl-4-(pyridin-4-yl)thieno[2,3-d]pyrimidine (compound) was used. 6-6) Instead of 6-bromo-4-(pyridin-4-yl)thieno[2,3-d]pyrimidine (compounds 1-6).
  • Compound 7 was prepared in a similar manner to Compound 1 prepared in Example 1, except that N-(2-methoxy-5-(4,4,5,5-tetramethyl-1,3,2) was used in step 6. -dioxaborolan-2-yl)pyridin-3-yl)-4-cyanobenzenesulfonamide instead of N-(2-methoxy-5-(4,4,5,5-tetramethyl) Base-1,3,2-dioxolane Alk-2-yl)pyridin-3-yl)-2,4-difluorobenzenesulfonamide.
  • Compound 8 was prepared in a similar manner to Compound 1 prepared in Example 1, except that N-(2-methoxy-5-(4,4,5,5-tetramethyl-1,3,2) was used in step 6. -dioxaborolan-2-yl)pyridin-3-yl)-4-(trifluoromethyl)benzenesulfonamide instead of N-(2-methoxy-5-(4,4,5, 5-Tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-3-yl)-2,4-difluorobenzenesulfonamide.
  • Compound 9 was prepared in a similar manner to Compound 1 prepared in Example 1, except that N-(2-methoxy-5-(4,4,5,5-tetramethyl-1,3,2) was used in step 6. -dioxaborolan-2-yl)pyridin-3-yl)-5-chlorothiophene-2-sulfonamide instead of N-(2-methoxy-5-(4,4,5,5- Tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-3-yl)-2,4-difluorobenzenesulfonamide.
  • N-(5-Bromo-2-methylpyridin-3-yl)-2,4-difluorobenzenesulfonamide (87 mg, 0.24 mmol), boronic acid pinacol ester (73 mg, 0.288 mmol) and anhydrous
  • a mixture of potassium acetate (94 mg, 0.96 mmol) in dioxane (6 mL) was degassed, then [1,1 '-bis(diphenylphosphino)ferrocene]palladium dichloride (17.6 mg, 0.024 mmol) was added to the mixture.
  • the resulting reaction mixture was degassed and backfilled with argon (three cycles), stirred at 100 ° C for 4 hours under an argon atmosphere, and then cooled to room temperature.
  • N-(5-Bromo-2-chloropyridin-3-yl)-4-fluorobenzenesulfonamide (88 mg, 0.24 mmol), boronic acid pinacol ester (73 mg, 0.288 mmol) and anhydrous potassium acetate (47 mg) , 0.48 mmol) of the mixture in dioxane (6 mL) was degassed, then [1,1 '-bis(diphenylphosphino)ferrocene]palladium dichloride (13 mg, 0.018 mmol) was added In the mixture. The resulting reaction mixture was degassed and backfilled with argon (three cycles) and stirred at 100 ° C for 3 hours under an argon atmosphere. The reaction mixture was cooled to room temperature and used directly in the next step.
  • N-(5-Bromo-2-chloropyridin-3-yl)-2,4-difluorobenzenesulfonamide 92 mg, 0.24 mmol
  • anhydrous potassium acetate 47 mg, 0.48 mmol
  • diboronic acid pinacol The mixture of ester (73 mg, 0.288 mmol) in dioxane (6 mL) was degassed then [1,1 '-bis(diphenylphosphino)ferrocene]palladium dichloride (9 mg, 0.012 mmol) ) is added to the mixture.
  • the resulting reaction mixture was degassed and backfilled with argon (three cycles), stirred at 100 ° C for 3 hours under an argon atmosphere, and then cooled to room temperature.
  • N-(5-Bromo-2-chloropyridin-3-yl)-5-chlorothiophene-2-sulfonamide (93 mg, 0.24 mmol), boronic acid pinacol ester (73 mg, 0.288 mmol) and potassium acetate (
  • a mixture of 47 mg, 0.48 mmol) in dioxane (6 mL) was degassed then [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (9 mg, 0.012 mmol).
  • the resulting reaction mixture was degassed and backfilled with argon (three cycles), stirred at 100 ° C for 3.5 hours under an argon atmosphere, and then cooled to room temperature.
  • the potency of the compounds of the invention against PI3K-[alpha] was assessed using an in vitro kinase assay.
  • the kinase activity of PI3K- ⁇ was determined by detecting the level of ADP produced in the kinase reaction using luciferase-based luminescence detection.
  • ADP-Glo TM kinase assay kit was purchased from Promega. All tests were performed at room temperature using a white OptiPlate TM -384 384-well plates.
  • PI3K-alpha kinase is from Crownbio.
  • the substrate was PIP2/PS (Invitrogen).
  • the kinase buffer included 50 mM Hepes (pH 7.5), 3 mM MgCl2, 100 mM NaCl, 1 nM EGTA, 0.03% CHAPS and 2 mM DTT.
  • a PI3K-alpha kinase cocktail was prepared by diluting PI3K-alpha kinase to 0.375 ng/ ⁇ L in kinase buffer.
  • the ATP/substrate mix included 5 ⁇ M PIP 2/PS and 25 ⁇ M ATP.
  • the test compound was diluted to 10 mM in 100% DMSO and then serially diluted 3 times into 10 different concentrations in 100% DMSO.
  • the compound was then diluted in 100% DMSO is diluted 20-fold in ddH 2 O in. 2 ⁇ L of the diluted compound solution and 4 ⁇ L of the ATP/substrate mixture were added to each well of a 384-well plate.
  • the reaction was started by adding 4 ⁇ L of PI3K ⁇ kinase mixture per well, and the final reaction volume was 10 ⁇ L, the ATP concentration was 10 ⁇ M, the PIP 2/PS concentration was 2 ⁇ M, and the PI3K- ⁇ kinase concentration was 0.15 ng/ ⁇ L.
  • the plate was capped and the reaction was allowed to proceed for 1 hour at room temperature, followed by the addition of 10 [mu]L of kinase Glo (TM) reagent per well to stop the reaction.
  • the plates were incubated for 40 min and then 20 [mu]L of Kinase Detection Reagent per well was added.
  • the plates were equilibrated in the dark for 30 min and then luminescence was read on an EnVision 2104 multi-label microplate reader.
  • the percent inhibition is calculated based on the following formula:
  • Inhibition % [1-(RLU compound -RLU min )/(RLU max -RLU min )] ⁇ 100
  • the compound is a luminescent RLU reading at a given compound concentration
  • luminescence readings of RLU min was added to the highest concentration (2.5uM) positive control compound, the luminescence in the case where RLU readings without addition of compounds of max.
  • IC 50 is calculated by using the compounds in Excel XLfit program.
  • the compound of the present invention has a significant inhibitory effect on PI3K- ⁇ , and the compound of the present invention can be used for the preparation of a medicament for treating and preventing cancer and tumor diseases.

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Abstract

本发明公开了一种取代的噻吩并嘧啶化合物，以及它们的制备方法和其作为PI3-K和/或mTOR激酶抑制剂的用途。该取代的噻吩并嘧啶化合物及其药学上可接受的盐可用于制备治疗和/或预防肿瘤疾病的药物。

Description

一种取代的噻吩并嘧啶化合物及其制备方法 技术领域
本发明涉及一种噻吩并嘧啶化合物，属于药物化学技术领域。
背景技术
抗肿瘤药物的研究与开发是当今生命科学中极富挑战性且意义重大的领域。近年来,随着分子生物学的飞速发展以及人们对癌症发生、发展、作用机制的进一步认识，恶性肿瘤细胞内的信号转导、细胞周期的调控、细胞凋亡的诱导、血管生成以及细胞与胞外基质的相互作用等各种基本过程正在被逐步阐明。因此寻找和发现选择性作用于特定靶点的高效、低毒、特异性强的新型抗肿瘤药物已成为当前药物研究开发的重要领域之一。由此产生了一个新的抗癌药物领域-分子靶向药物。
分子靶向药物是指针对细胞癌变过程的受体或转导过程中关键的酶，从分子水平抑制肿瘤生长的治疗模式。以肿瘤细胞的特征分子为靶点，在发挥抗肿瘤作用的同时，减少了对正常细胞的毒副作用。
磷酸肌醇3-激酶(PI3-K)催化磷脂酰肌醇(PI)第二信使PI(3)P、PI(3,4)P2及PI(3,4,5)P3(PIP3)的合成。在适当的细胞环境下，这三种脂质控制多种包括细胞生长、存活、分化及趋化性的生理学过程。癌症中的主要PI3-激酶同功异型物为由催化性(p110α)亚单元及转接子(p85)亚单元组成的Ⅰ类PI3-Kα。PI3-K/Akt/PTEN通路的数种组分涉及肿瘤形成，在包括细胞周期调节、增生、存活、细胞凋亡及运动的许多细胞过程的调控中发挥重要作用，且为诸如癌症、糖尿病及免疫性炎症等疾病的分子机制的重要组分。
因此，PI3-Kα为开发癌症药物的具有吸引力和前景的靶标，此类药剂将抑制癌细胞增生，且避免了细胞毒制剂对机体的损伤，及癌细胞对细胞毒制剂的耐受。人们需要新颖的PI3-Kα抑制剂，作为癌症治疗药物应用于癌症治疗领域。
发明内容
本发明提供一种取代的噻吩并嘧啶化合物，以及它们的制备方法和其作为PI3-K和/或mTOR激酶抑制剂的用途。
本发明的技术方案具体如下：
一种取代的噻吩并嘧啶化合物，具有如下式Ⅰ所示的结构：
Figure PCTCN2015095384-appb-000001
其中：
R1选自取代或未取代的芳基、取代或未取代的芳杂环基，所述芳杂环基的杂原子选自N、S和O；
R2选自氢、C1-C4烷基；
R3选自C1-C4烷氧基、C1-C4烷基、卤素(氟、氯、溴、碘)；
X为C或N；
当X为N时，R4不存在；
当X为C时，R4选自氢、卤素、取代或未取代的C1-C4烷基、取代或未取代的C1-C4烷氧基、取代或未取代的氨基、取代或未取代的磺酰基氨基。
式I化合物药学上可接受的盐也在本发明的保护范围内。
进一步的，R1选自取代或未取代的苯基、取代或未取代的吡啶基、取代或未取代的吡喃基、取代或未取代的哒嗪基、取代或未取代的嘧啶基、取代或未取代的吡嗪基、取代或未取代的哌嗪基、取代或未取代的呋喃基、取代或未取代的噻吩基、取代或未取代的吡咯基、取代或未取代的吡唑基、取代或未取代的咪唑基、取代或未取代的噁唑基、取代或未取代的噻唑基、取代或未取代的异噁唑基。
所述R1基团上带有取代基时，取代基的个数可以是1-5个，取代基选自卤素、氰基、三氟甲基、C1-C4烷氧基、C1-C4烷基、氨基和羟基。当R1为吡啶基时，所述取代的噻吩并嘧啶化合物为式II所示的化合物：
Figure PCTCN2015095384-appb-000002
上述式I和式II中，当X为C，R4为取代的C1-C4烷基、取代的C1-C4烷氧基、取代的氨基时，取代基选自卤素、氰基、三氟甲基、C1-C4烷氧基、C1-C4烷基、氨基和羟基。
当X为C，R4为取代或未取代的磺酰基氨基时，所述磺酰基氨基为-NH-SO2-R5；其中，R5选自取代或未取代的C1-C4烷基、取代或未取代的C3-C7环烷基、取代或未取代的苯基、取代或未取代的五元或六元芳杂环基，所述芳杂环的杂原子选自N、S和O。
进一步的，R5选自取代或未取代的C1-C4烷基、取代或未取代的C3-C7环烷基、取代或未取代的苯基、取代或未取代的吡啶基、取代或未取代的吡喃基、取代或未取代的哒嗪基、取代或未取代的嘧啶基、取代或未取代的吡嗪基、取代或未取代的哌嗪基、取代或未取代的呋喃基、取代或未取代的噻吩基、取代或未取代的吡咯基、取代或未取代的吡唑基、取代或未取代的咪唑基、取代或未取代的噁唑基、取代或未取代的噻唑基、取代或未取代的异噁唑基。
其中，所述的R5基团上带有取代基时，取代基的个数为1-5个，这些取代基选自卤素(氟、氯、溴、碘)、氰基、三氟甲基、C1-C4烷氧基、C1-C4烷基、氨基、羟基。
例如：R5可选自下列基团：
Figure PCTCN2015095384-appb-000003
具体的，式I所示化合物可选自下列化合物1～13：
Figure PCTCN2015095384-appb-000004
本发明还提供了式I所示的化合物的制备方法，其包括：使式A所示的化合物与式B所示的化合物在钯催化剂的存在下发生如下取代反应，从而得到式I所示的化合物：
Figure PCTCN2015095384-appb-000005
其中，R1，R2，R3，R4和X如前所定义；R6为
Figure PCTCN2015095384-appb-000006
上述制备方法中，所述的钯催化剂可以是四(三苯基膦)钯，二(三苯基膦)二氯化钯或[1,1’-双(二苯基膦基)二茂铁]二氯化钯。
上述制备方法中，式A所示的化合物可通过如下方法制备得到的：
使式G所示的化合物在钯催化剂的存在下与式H所示的化合物发生如下反应，从而得到式A所示的化合物：
Figure PCTCN2015095384-appb-000007
其中，R1和R2如前所定义；R7为
Figure PCTCN2015095384-appb-000008
其中，所述的钯催化剂可以是四(三苯基膦)钯，二(三苯基膦)二氯化钯或[1,1’-双(二苯基膦基)二茂铁]二氯化钯。
上述式G所示的化合物可通过如下方法制备得到的：
使式F所示的化合物先经氯化氢乙醚溶液处理，再与碘化钠反应得到式G所示的化合物：
Figure PCTCN2015095384-appb-000009
其中，R2如前所定义。
上述式F所示的化合物可通过如下方法制备得到的：
使式E所示的化合物与三氯氧磷，氯化亚砜和草酰氯等氯代试剂发生如下反应，得到式F所示的化合物：
Figure PCTCN2015095384-appb-000010
其中，R2如前所定义。
上述式E所示的化合物是通过如下方法制备得到的：
使式D所示的化合物与液溴或N-溴代丁二酰亚胺等溴代试剂发生如下反应，得到式E所示的化合物：
Figure PCTCN2015095384-appb-000011
其中，R2如前所定义。
上述式D所示的化合物可通过如下方法制备得到的：
使式C所示的化合物与醋酸甲脒或甲酰胺等试剂发生如下环化反应，得到式D所示的化合物：
Figure PCTCN2015095384-appb-000012
其中，R2如前所定义。
本发明说明书中二氯甲烷的英文缩写是DCM，N-甲基吡咯烷酮的英文缩写是NMP。
本发明取代的噻吩并嘧啶化合物是一种多激酶抑制剂，能同时抑制多种存在于细胞内和细胞表面的激酶，包括PI3-K和/或mTOR激酶。所述PI3-K优先指PI3-Kα。该化合物及其药学上可接受的盐可用于制备治疗和/或预防PI3-K和/或mTOR激酶引起的疾病的药物。所述药学上可接受的盐包括：盐酸盐、氢溴酸盐、硫酸盐、磷酸盐、甲磺酸盐、三氟甲磺酸盐、苯磺酸盐、对甲苯磺酸盐、1-萘磺酸盐、2-萘磺酸盐、乙酸盐、三氟乙酸盐、苹果酸盐、酒石酸盐、柠檬酸盐、乳酸盐、草酸盐、琥珀酸盐、富马酸盐、马来酸盐、苯甲酸盐、水杨酸盐、苯基乙酸盐、杏仁酸盐等。
发明还提供了一种治疗和/或预防PI3-K和/或mTOR激酶引起的疾病的药物组合物，所述的药物组合物中含有本发明所述的取代的噻吩并嘧啶化合物及药用辅料。优选地，所述的药物组合物为注射剂、口服制剂、透皮吸收剂或栓剂。
PI3-K和/或mTOR激酶引起的疾病例如癌症，例如：脑癌、成胶质细胞瘤、头颈癌、肺癌、黑色素瘤、肝癌、肾癌、急性白血病、慢性白血病、非小细胞肺癌、前列腺癌、甲状腺癌、皮肤癌、结肠癌、直肠癌、胰腺癌、卵巢癌、乳腺癌、骨髓异常增生综合症、食管癌、肉瘤、骨肉瘤或横纹肌瘤。
本发明所述的取代的噻吩并嘧啶化合物及其药学上可接受的盐 可用于制备治疗和/或预防肿瘤疾病的药物。
具体实施方式
以下通过具体实施例对本发明作进一步说明。在本发明说明书和权利要求中，化合物的命名都是依据化学结构式，如果表示同一化合物时化合物的命名与化学结构式不符，以化学结构式或化学反应式为准。
实施例1：N-(2-甲氧基-5-(4-(吡啶-4-基)噻吩并[2,3-d]嘧啶-6-基)吡啶-3-基)-2,4-二氟苯磺酰胺(化合物1)的合成
Figure PCTCN2015095384-appb-000013
步骤1：噻吩并[2,3-d]嘧啶-4-酚(化合物1-2)的合成
Figure PCTCN2015095384-appb-000014
将2-氨基噻吩-3-羧酸甲酯(1-1)(15.72g，100mmol)和醋酸甲脒(20.6g，200mmol)于N-甲基吡咯烷酮(80mL)中的混合物在135℃下氩气气氛中搅拌24小时。冷却至室温后，用水(400mL)稀释反应混合物，用乙酸乙酯(200mL×3)萃取。用水(200mL×3)和食盐水(200mL)洗涤合并的有机层，用无水硫酸钠干燥，过滤，浓缩得到粗产品(化合物1-2)为褐色固体(13g，产率85％)，不经进一步纯化直接用于下一步。
化合物1-2：1H NMR(400MHz,DMSO-d6)δ12.48(br s,1H),8.12(d,J＝2.7Hz,1H),7.58(d,J＝5.8Hz,1H),7.39(d,J＝5.8Hz,1H).
MS(ESI+)m/z 153.1[M+H]+.
步骤2：6-溴噻吩并[2,3-d]嘧啶-4-酚(化合物1-3)的合成
Figure PCTCN2015095384-appb-000015
向噻吩并[2,3-d]嘧啶-4-酚(化合物1-2)(13g，85.43mmol)于冰醋酸(250mL)中的混合物中缓慢加入液溴(8.75mL，27.3g，170.86mmol，2.0eq)，然后将产生的反应混合物在室温下搅拌3小时。将反应混合物分配于水(1000mL)和乙酸乙酯(300mL)之间，用饱和硫代硫酸钠水溶液淬灭残留的溴。抽滤收集产生的黄色固体，在50℃下烘干得到粗产物(13.40g)。将滤液转移到分液漏斗中，分离有机层，并用乙酸乙酯(300mL×2)萃取水层。用水(300mL×3)和食盐水(300mL)洗涤合并的有机层，用无水硫酸钠干燥，过滤，蒸干得到粗产物(2.80g)。合并两批次的粗产物(化合物1-3)(16.2g，产率82.4％)，不经进一步纯化直接用于下一步。
化合物1-3：1H NMR(400MHz,DMSO-d6)δ12.65(br s,1H),8.15(s,1H),7.56(s,1H).
MS(ESI+)m/z 230.9[M+H]+.
步骤3：6-溴-4-氯噻吩并[2,3-d]嘧啶(化合物1-4)的合成
Figure PCTCN2015095384-appb-000016
将6-溴噻吩并[2,3-d]嘧啶-4-酚(化合物1-3)(13.58g，58.77mmol)于三氯氧磷(70mL)中的混合物回流2小时。在真空中除去三氯氧磷后，将残留物倒入冰水(200mL)中，用饱和碳酸氢钠水溶液中和。用乙酸乙酯(100mL×3)萃取产生的混合物。用水(100mL×2)和食盐水(100mL)洗涤合并的有机层，用无水硫酸钠干燥，过滤，浓缩。残留物经快速柱色谱(硅胶，洗脱液：石油醚/乙酸乙酯＝50:1(体积比))纯化得到产物(化合物1-4)为淡黄色固体(12.5g，产率84.9％)。
化合物1-4：1H NMR(400MHz,DMSO-d6)δ8.96(s,1H),7.90(s,1H).
MS(ESI+)m/z 248.9[M+H]+.
步骤4：6-溴-4-碘噻吩并[2,3-d]嘧啶(化合物1-5)的合成
Figure PCTCN2015095384-appb-000017
将6-溴-4-氯噻吩并[2,3-d]嘧啶(化合物1-4)(11g，44.1mmol)于无水四氢呋喃(80mL)中的溶液以干燥氯化氢气鼓泡30分钟，然后在真空中除去挥发物。向残留物中加入无水二氧六环(330mL)和碘化钠(33.0g，200.5mmol，5eq)，将产生的混合物在110℃下氩气气氛中回流24小时。在真空中除去二氧六环，用水(300mL)稀释残留物，用饱和硫代硫酸钠(10mL)处理。用乙酸乙酯(100mL×3)萃取产生的混合物。用水(100mL×2)和食盐水(100mL)洗涤合并的有机层，用无水硫酸钠干燥，过滤，浓缩。残留物经快速柱色谱(硅胶，洗脱液：石油醚/乙酸乙酯＝50:1(体积比))纯化得到产物(化合物1-5)为淡黄色固体(10g，66％)。
化合物1-5：1H NMR(400MHz,DMSO-d6)δ8.75(s,1H),7.65(s,1H).
MS(ESI+)m/z 340.8,342.8[M+H]+.
步骤5：6-溴-4-(吡啶-4-基)噻吩并[2,3-d]嘧啶(化合物1-6)的合成
Figure PCTCN2015095384-appb-000018
将6-溴-4-碘噻吩并[2,3-d]嘧啶(化合物1-5)(3.41g,10mmol)，吡啶-4-硼酸(1.29g,10.5mmol,1.05eq)和2N碳酸钾(15mL,30mmol,3.0eq)于二氧六环(75mL)中的混合物脱气，然后加入二(三苯基膦)二氯化钯(351mg,0.5mmol,0.05eq)。将产生的反应混合物脱气并回充氩气(三个 循环)，然后在100℃下氩气气氛中搅拌16小时。在真空中浓缩反应混合物，用水(100mL)稀释，用乙酸乙酯(50mL×3)萃取。用水(50mL×2)和食盐水(50mL)洗涤合并的有机层，用无水硫酸钠干燥，过滤，浓缩。残留物经快速柱色谱(硅胶，洗脱液：石油醚/乙酸乙酯＝5:1(体积比))纯化得到产物(化合物1-6)为淡黄色固体(0.90g，31％)。
化合物1-6：1H NMR(400MHz,DMSO-d6)δ9.23(s,1H),8.83(dd,J＝4.4,1.6Hz,2H),8.05(s,1H),7.96(dd,J＝4.4,1.6Hz,2H).
MS(ESI+)m/z 291.9[M+H]+.
步骤6：N-(2-甲氧基-5-(4-(吡啶-4-基)噻吩并[2,3-d]嘧啶-6-基)吡啶-3-基)-2,4-二氟苯磺酰胺(化合物1)的合成
Figure PCTCN2015095384-appb-000019
将6-溴-4-(吡啶-4-基)噻吩并[2,3-d]嘧啶(化合物1-6)(102mg,0.35mmol)，N-(2-甲氧基-5-(4,4,5,5-四甲基-1,3,2-二氧杂环戊硼烷-2-基)吡啶-3-基)-2,4-二氟苯磺酰胺(179mg,0.42mmol,1.2eq)和2N碳酸钾(0.70mL,1.4mmol,4.0eq)于二氧六环(5mL)中的混合物脱气，然后加入二(三苯基膦)二氯化钯(26mg,0.035mmol,0.1eq)。将产生的反应混合物脱气并回充氩气(三个循环)，然后在100℃下氩气气氛中搅拌6小时。在真空中浓缩反应混合物，用水(30mL)稀释，用乙酸乙酯(30mL×3)萃取。用水(30mL)和食盐水(30mL)洗涤合并的有机层，用无水硫酸钠干燥，过滤，浓缩。残留物经柱色谱(硅胶，洗脱液：DCM/甲醇＝100:1(体积比))纯化得到产物为黄色固体(110mg，产率为61％)。
化合物1：1H-NMR(400MHZ,DMSO-d6):δ10.44(s,1H),9.21(s,1H),8.86(d,J＝6.0Hz,2H),8.60(d,J＝2.0Hz,1H),8.22–8.13(m,2H),8.04(dd,J＝6.0Hz,2H),7.77(td,J＝8.4,6.8Hz,1H),7.59(t,J＝8.8Hz,1H), 7.22(td,J＝8.4,2.4Hz,1H),3.66(s,3H).
MS(ESI+)m/z 512.0[M+H]+.
实施例2：N-(2-甲氧基-5-(4-(吡啶-4-基)噻吩并[2,3-d]嘧啶-6-基)吡啶-3-基)甲磺酰胺(化合物2)的合成
Figure PCTCN2015095384-appb-000020
以类似于实施例1中制备化合物1的方式制备化合物2，除了在步骤6使用N-(2-甲氧基-5-(4,4,5,5-四甲基-1,3,2-二氧杂环戊硼烷-2-基)吡啶-3-基)甲磺酰胺代替N-(2-甲氧基-5-(4,4,5,5-四甲基-1,3,2-二氧杂环戊硼烷-2-基)吡啶-3-基)-2,4-二氟苯磺酰胺。
化合物2：1H NMR(400MHz,DMSO-d6)δ9.50(s,1H),9.22(s,1H),8.86(d,J＝6.0Hz,2H),8.61(d,J＝2.3Hz,1H),8.15(s,1H),8.12(d,J＝2.3Hz,1H),8.04(d,J＝6.0Hz,2H),4.00(s,3H),3.11(s,3H).
MS(ESI+)m/z 414.0[M+H]+.
实施例3：6-(6-甲氧基吡啶-3-基)-4-(吡啶-4-基)噻吩并[2,3-d]嘧啶(化合物3)的合成
Figure PCTCN2015095384-appb-000021
以类似于实施例1中制备化合物1的方式制备化合物3，除了在步骤6使用2-甲氧基吡啶-5-硼酸代替N-(2-甲氧基-5-(4,4,5,5-四甲基-1,3,2-二氧杂环戊硼烷-2-基)吡啶-3-基)-2,4-二氟苯磺酰胺。
化合物3：1H NMR(400MHz,DMSO)δ9.21(s,1H),8.85(d,J＝6.0Hz,2H),8.76(d,J＝2.4Hz,1H),8.34(dd,J＝8.8,2.4Hz,1H),8.15(s,1H),8.05(d,J＝6.0Hz,2H),7.00(d,J＝8.8Hz,1H),3.94(s,3H).
MS(ESI+)m/z 321.1[M+H]+.
实施例4：6-(2-甲氧基嘧啶-5-基)-4-(吡啶-4-基)噻吩并[2,3-d]嘧啶(化合物4)的合成
Figure PCTCN2015095384-appb-000022
以类似于实施例1中制备化合物1的方式制备化合物4，除了在步骤6使用2-甲氧基嘧啶-5-硼酸代替N-(2-甲氧基-5-(4,4,5,5-四甲基-1,3,2-二氧杂环戊硼烷-2-基)吡啶-3-基)-2,4-二氟苯磺酰胺。
化合物4：1H NMR(400MHz,DMSO-d6)δ9.24(s,1H),9.21(s,2H),8.86(d,J＝6.1Hz,2H),8.29(s,1H),8.05(d,J＝6.1Hz,2H),4.01(s,3H).
实施例5：6-(5-氟-6-甲氧基吡啶-3-基)-4-(吡啶-4-基)噻吩并[2,3-d]嘧啶(化合物5)的合成
Figure PCTCN2015095384-appb-000023
以类似于实施例1中制备化合物1的方式制备化合物5，除了在步骤6使用2-甲氧基-3-氟吡啶-5-硼酸代替N-(2-甲氧基-5-(4,4,5,5-四甲基-1,3,2-二氧杂环戊硼烷-2-基)吡啶-3-基)-2,4-二氟苯磺酰胺。
化合物5：1H NMR(400MHz,DMSO-d6)δ9.22(s,1H),8.86(dd,J＝4.4,1.6Hz,2H),8.53(d,J＝2.1Hz,1H),8.49(dd,J＝11.6,2.1Hz,1H),8.22(s,1H),8.04(dd,J＝4.4,1.6Hz,2H),4.02(s,3H).
MS(ESI+)m/z 339.0[M+H]+.
实施例6：N-(2-甲氧基-5-(5-甲基-4-(吡啶-4-基)噻吩并[2,3-d]嘧啶-6-基)吡啶-3-基)-2,4-二氟苯磺酰胺(化合物6)的合成
Figure PCTCN2015095384-appb-000024
步骤1：5-甲基噻吩并[2,3-d]嘧啶-4-酚(6-2)的合成
Figure PCTCN2015095384-appb-000025
以类似于实施例1中的步骤1制备化合物1-2的方式制备化合物6-2，除了使用2-氨基-4-甲基噻吩-3-羧酸甲酯(6-1)代替2-氨基噻吩-3-羧酸甲酯(1-1)。
化合物6-2：1H NMR(400MHz,DMSO-d6):δ12.31(br s,1H),8.05(s,1H),7.14(s,1H),2.46(s,3H).
步骤2：6-溴-5-甲基噻吩并[2,3-d]嘧啶-4-酚(化合物6-3)的合成
Figure PCTCN2015095384-appb-000026
以类似于实施例1中步骤2制备化合物1-3的方式制备化合物6-3，除了使用5-甲基噻吩并[2,3-d]嘧啶-4-酚(化合物6-2)代替噻吩并[2,3-d]嘧啶-4-酚(化合物1-2)。
化合物6-3：1H NMR(400MHz,DMSO-d6):δ12.57(s,1H),8.11(s,1H),2.43(s,3H).
步骤3：6-溴-4-氯-5-甲基噻吩并[2,3-d]嘧啶(化合物6-4)的合成
Figure PCTCN2015095384-appb-000027
以类似于实施例1中步骤3制备化合物1-4的方式制备化合物6-4，除了使用6-溴-5-甲基噻吩并[2,3-d]嘧啶-4-酚(化合物6-3)代替6-溴噻吩并[2,3-d]嘧啶-4-酚(化合物1-3)。
化合物6-4：1H NMR(400MHz,DMSO-d6):δ8.92(s,1H),2.60(s,3H).
步骤4：6-溴-4-碘-5-甲基噻吩并[2,3-d]嘧啶(化合物6-5)的合成
Figure PCTCN2015095384-appb-000028
以类似于实施例1中步骤4制备化合物1-5的方式制备化合物6-5，除了使用6-溴-4-氯-5-甲基噻吩并[2,3-d]嘧啶(化合物6-4)代替6-溴-4-氯噻吩并[2,3-d]嘧啶(化合物1-4)。
化合物6-5：1H NMR(400MHz,DMSO-d6):δ8.69(s,1H),2.61(s,3H).
步骤5：6-溴-5-甲基-4-(吡啶-4-基)噻吩并[2,3-d]嘧啶(化合物6-6)的合成
Figure PCTCN2015095384-appb-000029
以类似于实施例1中步骤5制备化合物1-6的方式制备化合物6-6，除了使用6-溴-4-碘-5-甲基噻吩并[2,3-d]嘧啶(化合物6-5)代替6-溴-4-碘噻吩并[2,3-d]嘧啶(化合物1-5)。
化合物6-6：1H NMR(400MHz,DMSO-d6):δ9.16(s,1H),8.78(d,J＝6.0Hz,2H),7.63(d,J＝6.0Hz,2H),1.88(s,3H)
步骤6：N-(2-甲氧基-5-(5-甲基-4-(吡啶-4-基)噻吩并[2,3-d]嘧啶-6-基)吡啶-3-基)-2,4-二氟苯磺酰胺(化合物6)的合成
Figure PCTCN2015095384-appb-000030
以类似于实施例1中步骤6制备化合物1的方式制备化合物6，除了使用6-溴-5-甲基-4-(吡啶-4-基)噻吩并[2,3-d]嘧啶(化合物6-6)代替6-溴-4-(吡啶-4-基)噻吩并[2,3-d]嘧啶(化合物1-6)。
化合物6：1H NMR(400MHz,DMSO-d6):δ10.48(s,1H),9.19(s,1H),8.79(d,J＝6.0Hz,2H),8.22(d,J＝1.6Hz,1H),7.82–7.71(m,2H),7.67(d,J＝6.0Hz,2H),7.56(t,J＝10Hz,1H),7.23(td,J＝8.4,1.6Hz,1H),3.71(s,3H),1.85(s,3H).
实施例7：N-(2-甲氧基-5-(4-(吡啶-4-基)噻吩并[2,3-d]嘧啶-6-基)吡啶-3-基)-4-氰基苯磺酰胺(化合物7)的合成
Figure PCTCN2015095384-appb-000031
以类似于实施例1中制备化合物1的方式制备化合物7，除了在步骤6使用N-(2-甲氧基-5-(4,4,5,5-四甲基-1,3,2-二氧杂环戊硼烷-2-基)吡啶-3-基)-4-氰基苯磺酰胺代替N-(2-甲氧基-5-(4,4,5,5-四甲基-1,3,2-二氧杂环戊硼 烷-2-基)吡啶-3-基)-2,4-二氟苯磺酰胺。
化合物7：1H NMR(400MHZ,DMSO-d6):δ10.46(s,1H),9.22(s,1H),8.87(dd,J＝4.8,2.0Hz,2H),8.55(s,1H),8.1–8.02(m,6H),7.90(d,J＝8.5Hz,2H),3.61(s,3H).
MS(ESI+)m/z 501.0[M+H]+.
实施例8：N-(2-甲氧基-5-(4-(吡啶-4-基)噻吩并[2,3-d]嘧啶-6-基)吡啶-3-基)-4-(三氟甲基)苯磺酰胺(化合物8)的合成
Figure PCTCN2015095384-appb-000032
以类似于实施例1中制备化合物1的方式制备化合物8，除了在步骤6使用N-(2-甲氧基-5-(4,4,5,5-四甲基-1,3,2-二氧杂环戊硼烷-2-基)吡啶-3-基)-4-(三氟甲基)苯磺酰胺代替N-(2-甲氧基-5-(4,4,5,5-四甲基-1,3,2-二氧杂环戊硼烷-2-基)吡啶-3-基)-2,4-二氟苯磺酰胺。
化合物8：1H NMR(400MHZ,DMSO-d6):δ10.40(s,1H),9.22(s,1H),8.86(dd,J＝4.5,1.6Hz,2H),8.60(d,J＝2.4Hz,1H),8.16(s,1H),8.15(d,J＝2.4Hz,1H),8.05(dd,J＝4.5,1.6Hz,2H),7.99(d,J＝8.4Hz,2H),7.94(d,J＝8.4Hz,2H),3.56(s,3H).
MS(ESI+)m/z 544.0[M+H]+.
实施例9：N-(2-甲氧基-5-(4-(吡啶-4-基)噻吩并[2,3-d]嘧啶-6-基)吡啶-3-基)-5-氯噻吩-2-磺酰胺(化合物9)的合成
Figure PCTCN2015095384-appb-000033
以类似于实施例1中制备化合物1的方式制备化合物9，除了在步骤6使用N-(2-甲氧基-5-(4,4,5,5-四甲基-1,3,2-二氧杂环戊硼烷-2-基)吡啶-3-基)-5-氯噻吩-2-磺酰胺代替N-(2-甲氧基-5-(4,4,5,5-四甲基-1,3,2-二氧杂环戊硼烷-2-基)吡啶-3-基)-2,4-二氟苯磺酰胺。
化合物9：1H NMR(400MHZ,DMSO-d6):δ10.48(s,1H),9.23(s,1H),8.87(dd,J＝4.5,1.5Hz,2H),8.65(d,J＝2.0Hz,1H),8.16(s,1H),8.14(d,J＝2.0Hz,1H),8.05(dd,J＝4.5,1.6Hz,2H),7.41(d,J＝4.1Hz,1H),7.25(d,J＝4.1Hz,1H),3.77(s,3H).
MS(ESI+)m/z 516.0[M+H]+.
实施例10：N-(2-甲基-5-(4-(吡啶-4-基)噻吩并[2,3-d]嘧啶-6-基)吡啶-3-基)-2,4-二氟苯磺酰胺(化合物10)的合成
Figure PCTCN2015095384-appb-000034
将N-(5-溴-2-甲基吡啶-3-基)-2,4-二氟苯磺酰胺(87mg,0.24mmol)，联硼酸频哪醇酯(73mg,0.288mmol)和无水醋酸钾(94mg,0.96mmol)在二氧六环(6mL)中的混合物脱气，然后将[1,1'-双(二苯基膦基)二茂铁]二氯化钯(17.6mg,0.024mmol)加到混合物中。将产生的反应混合物脱气并回充氩气(三个循环)，在100℃下氩气气氛中搅拌4小时，然后冷却至室温。
向上述反应混合物中加入6-溴-4-(吡啶-4-基)噻吩并[2,3-d]嘧啶(58mg,0.2mmol)，2N碳酸钾水溶液(300μL,0.6mmol)和[1,1'-双(二苯基膦基)二茂铁]二氯化钯(14.6mg,0.02mmol)。将产生的反应混合物脱气并回充氩气(三个循环)，在100℃下氩气气氛中搅拌4小时。将反应混合物冷却至室温，用水(30mL)稀释，用乙酸乙酯萃取(30mL×3)。用食盐水(30mL)洗涤合并的有机层，用无水硫酸钠干燥，过滤，浓缩。残留物经快速柱色谱(硅胶，洗脱液：二氯甲烷/10％氨水甲醇溶液＝100:1(体积比))纯化得到产物10为淡黄色固体(62mg，产率为62.6％)。
化合物10：1H NMR(400MHz,DMSO-d6)δ10.59(br s,1H),9.24(s,1H),8.95(d,J＝1.6Hz,1H),8.87(dd,J＝4.4,1.6Hz,2H),8.22(s,1H),8.04(dd,J＝4.4,1.6Hz,2H),7.86(d,J＝2.0Hz,1H),7.80(td,J＝8.4,6.3Hz,1H),7.67–7.58(m,1H),7.26(td,J＝8.4,2.0Hz,1H),2.33(s,3H).
MS(ESI+)m/z 496.0[M+H]+.
实施例11：N-(2-氯-5-(4-(吡啶-4-基)噻吩并[2,3-d]嘧啶-6-基)吡啶-3-基)-4-氟苯磺酰胺(化合物11)的合成
Figure PCTCN2015095384-appb-000035
将N-(5-溴-2-氯吡啶-3-基)-4-氟苯磺酰胺(88mg,0.24mmol)，联硼酸频哪醇酯(73mg,0.288mmol)和无水醋酸钾(47mg,0.48mmol)在二氧六环(6mL)中的混合物脱气，然后将[1,1'-双(二苯基膦基)二茂铁]二氯化钯(13mg,0.018mmol)加到混合物中。将产生的反应混合物脱气并回充氩气(三个循环)，在100℃下氩气气氛中搅拌3小时。将反应混合物冷却至室温，直接用于下一步。
向上述反应混合物中加入6-溴-4-(吡啶-4-基)噻吩并[2,3-d]嘧啶(58mg, 0.2mmol)，2N碳酸钾水溶液(300μL,0.6mmol)和[1,1'-双(二苯基膦基)二茂铁]二氯化钯(14mg,0.019mmol)。将产生的反应混合物脱气并回充氩气(三个循环)，在100℃下氩气气氛中搅拌3.5小时。将反应混合物冷却至室温，用水(30mL)稀释，用乙酸乙酯萃取(30mL×3)。用食盐水(30mL)洗涤合并的有机层，用无水硫酸钠干燥，过滤，浓缩。残留物经快速柱色谱(硅胶，洗脱液：二氯甲烷/10％氨水甲醇溶液＝80:1(体积比))纯化得到产物11为白色固体(32mg，产率为32.1％)。
化合物11：1H NMR(400MHz,DMSO-d6)δ10.64(br s,1H),9.27(s,1H),8.90(d,J＝2.4Hz,1H),8.88(dd,J＝4.4,1.6Hz,2H),8.34(s,1H),8.16(d,J＝2.3Hz,1H),8.05(dd,J＝4.4,1.6Hz,2H),7.88–7.77(m,2H),7.45(t,J＝8.8Hz,2H).
MS(ESI+)m/z 497.9[M+H]+.
实施例12：N-(2-氯-5-(4-(吡啶-4-基)噻吩并[2,3-d]嘧啶-6-基)吡啶-3-基)-2,4-二氟苯磺酰胺(化合物12)的合成
Figure PCTCN2015095384-appb-000036
将N-(5-溴-2-氯吡啶-3-基)-2,4-二氟苯磺酰胺(92mg,0.24mmol)，无水醋酸钾(47mg,0.48mmol)和联硼酸频哪醇酯(73mg,0.288mmol)在二氧六环(6mL)中的混合物脱气，然后将[1,1'-双(二苯基膦基)二茂铁]二氯化钯(9mg,0.012mmol)加到混合物中。将产生的反应混合物脱气并回充氩气(三个循环)，在100℃下氩气气氛中搅拌3小时，然后冷却至室温。
向上述反应混合物中加入6-溴-4-(吡啶-4-基)噻吩并[2,3-d]嘧啶(58mg,0.2mmol)，2N碳酸钾水溶液(300μL,0.6mmol)和[1,1'-双(二苯基膦基) 二茂铁]二氯化钯(14mg,0.019mmol)。将产生的反应混合物脱气并回充氩气(三个循环)，在100℃下氩气气氛中搅拌4小时。将反应混合物冷却至室温，用水(30mL)稀释，用乙酸乙酯(30mL×3)萃取。用食盐水(30mL)洗涤合并的有机层，用无水硫酸钠干燥，过滤，浓缩。残留物经快速柱色谱(硅胶，洗脱液：二氯甲烷/10％氨水甲醇溶液＝100:1(体积比))纯化得到产物12为黄色固体(28.7mg，产率为27.8％)。
化合物12：1H NMR(300MHz,DMSO-d6)δ11.01(br s,1H),9.27(s,1H),8.91(d,J＝2.0Hz,,1H),8.88(dd,J＝4.4,1.6Hz,2H),8.37(s,1H),8.29(d,J＝2.0Hz,1H),8.05(dd,J＝4.4,1.6Hz,2H),7.82(td,J＝8.4,6.2Hz,1H),7.65–7.55(m,1H),7.25(td,J＝8.4,2.3Hz,1H).
MS(ESI+)m/z 515.9[M+H]+.
实施例13：N-(2-氯-5-(4-(吡啶-4-基)噻吩并[2,3-d]嘧啶-6-基)吡啶-3-基)-5-氯噻吩-2-磺酰胺(化合物13)的合成
Figure PCTCN2015095384-appb-000037
将N-(5-溴-2-氯吡啶-3-基)-5-氯噻吩-2-磺酰胺(93mg,0.24mmol)，联硼酸频哪醇酯(73mg,0.288mmol)和醋酸钾(47mg,0.48mmol)于二氧六环(6mL)中的混合物脱气，然后加入[1,1'-双(二苯基膦基)二茂铁]二氯化钯(9mg,0.012mmol)。将产生的反应混合物脱气并回充氩气(三个循环)，在100℃下氩气气氛中搅拌3.5小时，然后冷却至室温。
向上述的反应混合物中加入6-溴-4-(吡啶-4-基)噻吩并[2,3-d]嘧啶(59mg,0.2mmol)，2N碳酸钾水溶液(300μL,0.6mmol)和[1,1'-双(二苯基膦基)二茂铁]二氯化钯(12mg,0.0164mmol)。将反应混合物脱气并回充氩气(三个循环)，在100℃下氩气气氛中反应6小时。冷却至室温后， 在真空中除去挥发物。用水(30mL)稀释残留物，用乙酸乙酯(30mL×3)萃取。用水(30mL)和食盐水(30mL)洗涤合并的有机层，用无水硫酸钠干燥，过滤，浓缩。残留物经快速柱色谱(硅胶，洗脱液：二氯甲烷/甲醇＝80:1(体积比))纯化得到产物13为白色固体(33mg，产率为31.7％)。
化合物13：1H NMR(400MHz,DMSO-d6)δ9.27(s,1H),8.93(d,J＝2.3Hz,1H),8.88(d,J＝6.0Hz,2H),8.36(s,1H),8.22(d,J＝2.3Hz,1H),8.06(d,J＝6.0Hz,2H),7.44(d,J＝4.1Hz,1H),7.28(d,J＝4.1Hz,1H).
MS(ESI+)m/z 521.8[M+H]+.
实验例：
实验方法：生物化学检测PI3K-α活性
使用体外激酶检测评估本发明的化合物对PI3K-α的效能。使用基于荧光素酶的冷光检测，通过检测激酶反应中所产生的ADP水平来测定PI3K-α的激酶活性。ADP-GloTM激酶检测试剂盒购自Promega公司。所有检测均在室温下进行，使用OptiPlateTM-384白色384孔板。PI3K-α激酶来自Crownbio。底物为PIP2/PS(Invitrogen)。激酶缓冲液包括50mM Hepes(pH 7.5)，3mM MgCl2，100mM NaCl，1nM EGTA，0.03％CHAPS和2mM DTT。通过在激酶缓冲液将PI3K-α激酶稀释为0.375ng/μL配制PI3K-α激酶混合液。ATP/底物混合液包括5μM PIP2/PS和25μM ATP。将待测化合物在100％DMSO中稀释至10mM，然后在100％DMSO中按3倍连续稀释成10个不同浓度。然后将稀释于100％DMSO中的化合物在ddH2O中稀释20倍。将2μL稀释的化合物溶液和4μL的ATP/底物混合液加到384孔板的各孔中。通过加入每孔4μL的PI3Kα激酶混合液开始反应，最终反应体积为10μL，ATP浓度为10μM，PIP2/PS浓度为2μM，PI3K-α激酶浓度为0.15ng/μL。将板盖住并将反应在室温进行1小时，随后加入每孔10μL的激酶GloTM试剂终止反应。将板孵育40min，然后加入每孔20μL激酶检测试剂。将板在暗处平衡30min，随后在EnVision 2104多标记微孔板检测仪读板器上读取冷光。
抑制百分率基于以下公式计算：
抑制％＝[1-(RLU化合物-RLUmin)/(RLUmax-RLUmin)]×100
其中RLU化合物为给定化合物浓度下的冷光读数，RLUmin为加入最高浓度(2.5uM)阳性对照化合物的情况下的冷光读数，RLUmax为不加入化合物的情况下的冷光读数。通过使用Excel中XLfit程序中来计算化合物的IC50。
实验结果见表1。
表1
Figure PCTCN2015095384-appb-000038
Figure PCTCN2015095384-appb-000039
Figure PCTCN2015095384-appb-000040
实验结论：从表1的数据可以看出，本发明的式I化合物对PI3K-α均具有明显的抑制作用。其中，化合物3和4的IC50值达到1000-5000nM；化合物2、6、8的IC50值达到10-100nM；化合物1、7、9、10的IC50值达到1-10nM；化合物11、12、13的IC50值达到0.1-1nM。
从上述实验中可以看出，本发明化合物对PI3K-α具有明显的抑制作用，本发明化合物可用于制备治疗和预防癌症、肿瘤疾病的药物。

Claims (10)

  1. 式I所示的化合物或其药学上可接受的盐：
    Figure PCTCN2015095384-appb-100001
    其中，
    R1选自取代或未取代的芳基、取代或未取代的芳杂环基，其中所述芳杂环基的杂原子选自N、S和O；
    R2选自氢和C1-C4烷基；
    R3选自C1-C4烷氧基、C1-C4烷基和卤素；
    X为C或N；
    当X为N时，R4不存在；
    当X为C时，R4选自氢、卤素、取代或未取代的C1-C4烷基、取代或未取代的C1-C4烷氧基、取代或未取代的氨基、取代或未取代的磺酰基氨基。
  2. 根据权利要求1所述的化合物或其药学上可接受的盐，其特征在于，R1选自取代或未取代的苯基、取代或未取代的吡啶基、取代或未取代的吡喃基、取代或未取代的哒嗪基、取代或未取代的嘧啶基、取代或未取代的吡嗪基、取代或未取代的哌嗪基、取代或未取代的呋喃基、取代或未取代的噻吩基、取代或未取代的吡咯基、取代或未取代的吡唑基、取代或未取代的咪唑基、取代或未取代的噁唑基、取代或未取代的噻唑基，以及取代或未取代的异噁唑基。
  3. 根据权利要求2所述的化合物或其药学上可接受的盐，其特征在于，所述化合物为式II所示的化合物：
    Figure PCTCN2015095384-appb-100002
  4. 根据权利要求1或2所述的化合物或其药学上可接受的盐，其特征在于，所述R1基团上带有取代基时，取代基的个数为1-5个，取代基选自卤素、氰基、三氟甲基、C1-C4烷氧基、C1-C4烷基、氨基和羟基。
  5. 根据权利要求1或3所述的化合物或其药学上可接受的盐，其特征在于，当X为C，R4为取代的C1-C4烷基、取代的C1-C4烷氧基、取代的氨基时，取代基选自卤素、氰基、三氟甲基、C1-C4烷氧基、C1-C4烷基、氨基和羟基。
  6. 根据权利要求1或3所述的化合物或其药学上可接受的盐，其特征在于，当X为C，R4为取代或未取代的磺酰基氨基时，所述磺酰基氨基为-NH-SO2-R5；其中，R5选自取代或未取代的C1-C4烷基、取代或未取代的C3-C7环烷基、取代或未取代的苯基、取代或未取代的五元或六元芳杂环基，所述芳杂环的杂原子选自N、S和O。
  7. 根据权利要求6所述的化合物或其药学上可接受的盐，其特征在于，R5选自取代或未取代的C1-C4烷基、取代或未取代的C3-C7环烷基、取代或未取代的苯基、取代或未取代的吡啶基、取代或未取代的吡喃基、取代或未取代的哒嗪基、取代或未取代的嘧啶基、取代或未取代的吡嗪基、取代或未取代的哌嗪基、取代或未取代的呋喃基、取代或未取代的噻吩基、取代或未取代的吡咯基、取代或未取代的吡唑基、取代或未取代的咪唑基、取代或未取代的噁唑基、取代或未取代的噻唑基、取代或未取代的异噁唑 基。
  8. 根据权利要求7所述的化合物或其药学上可接受的盐，其特征在于，所述R5基团上带有取代基时，取代基的个数为1-5个，取代基选自卤素、氰基、三氟甲基、C1-C4烷氧基、C1-C4烷基、氨基和羟基。
  9. 根据权利要求8所述的化合物或其药学上可接受的盐，其特征在于，R5选自下列基团之一：
    Figure PCTCN2015095384-appb-100003
  10. 根据权利要求1所述的化合物或其药学上可接受的盐，其特征在于，所述式I所示的化合物选自下列化合物之一：
    Figure PCTCN2015095384-appb-100004
    Figure PCTCN2015095384-appb-100005
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