CN106496232B - A kind of 1- with anti-tumor activity (2- oxinanes) -1H- pyrazolos [3,4-d] pyrimidines and preparation method thereof - Google Patents

Abstract

Translated fromChinese

本发明公开了一种具有抗肿瘤活性的1‑(2‑四氢吡喃)‑1H‑吡唑并[3,4‑d]嘧啶类化合物，其通式(Ⅰ)如下。本发明也公开了该化合物的合成方法，以及含所述化合物的药用组合物。本发明还公开了该药物组合物在治疗因蛋白激酶活性异常所引起疾病的应用。本发明开发出一种结构新颖，具有显著抗肿瘤活性的吡唑并[3,4‑d]嘧啶类化合物，该化合物可以用作为BRAF/VEGFR‑2激酶的双靶点抑制剂，在治疗因蛋白激酶活性异常所引起疾病方面有良好的效果和广阔的应用前景。The invention discloses a 1-(2-tetrahydropyran)-1H-pyrazolo[3,4-d]pyrimidine compound with antitumor activity, the general formula (I) of which is as follows. The invention also discloses a synthesis method of the compound and a pharmaceutical composition containing the compound. The invention also discloses the application of the pharmaceutical composition in treating diseases caused by abnormal protein kinase activity. The present invention develops a pyrazolo[3,4-d]pyrimidine compound with novel structure and significant anti-tumor activity. It has good effect and broad application prospect in diseases caused by abnormal protein kinase activity.

Description

Translated fromChinese

一种具有抗肿瘤活性的1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶类化合物及其制备方法A 1-(2-tetrahydropyran)-1H-pyrazolo[3,4-d] with antitumor activityPyrimidine compounds and their preparation methods

技术领域technical field

本发明涉及一种具有抗肿瘤活性的1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶类化合物及其制备方法，属于医药领域。The invention relates to a 1-(2-tetrahydropyran)-1H-pyrazolo[3,4-d]pyrimidine compound with antitumor activity and a preparation method thereof, belonging to the field of medicine.

背景技术Background technique

恶性肿瘤的治疗长期以来是一个世界性的难题。以往对肿瘤的治疗是通过发现肿瘤并破坏来实现,现在随着对细胞信号传导途径研究的不断深入，人们对肿瘤细胞内部的癌基因及抗癌基因的作用了解得越来越深入,使得针对肿瘤的特异性分子靶点设计新的抗肿瘤药物成为可能。现在已知，促分裂素原活化蛋白激酶(MAPK)信号通路与肿瘤细胞的增殖、分化、迁移和凋亡有关。BRAF是RAF三个亚型之一，是MAPK通路重要的转导因子，且BRAF在3个亚型中突变频率最高，大约7％～8％的人类肿瘤发生BRAF突变，ARAF和CRAF则较少发生突变。黑色素瘤中BRAF突变率最高，约40％～68％恶性(转移性)黑色素瘤发生BRAF突变。该突变是第一个在黑色素瘤中被发现的高频率突变基因，目前已经成为黑色素瘤的治疗靶标。The treatment of malignant tumors has long been a worldwide problem. In the past, the treatment of tumors was achieved by discovering and destroying tumors. Now, with the deepening of research on cell signal transduction pathways, people have a deeper understanding of the role of oncogenes and anti-cancer genes inside tumor cells, making it possible to target It is possible to design new anti-tumor drugs based on tumor-specific molecular targets. It is now known that the mitogen-activated protein kinase (MAPK) signaling pathway is related to the proliferation, differentiation, migration and apoptosis of tumor cells. BRAF is one of the three subtypes of RAF and an important transduction factor of the MAPK pathway, and BRAF has the highest mutation frequency among the three subtypes, about 7% to 8% of human tumors have BRAF mutations, while ARAF and CRAF are less Mutation occurs. The BRAF mutation rate is the highest in melanoma, and about 40% to 68% of malignant (metastatic) melanomas have BRAF mutations. This mutation is the first highly mutated gene found in melanoma, and has now become a therapeutic target for melanoma.

血管内皮细胞生长因子(vascular endothelial growth factor，VEGF)及其受体VEGFR(VEGF receptor)所介导的“出芽式血管生成(sprouting angiogenesis)”，在生理及病理性血管生成中均扮演着重要的角色，尤其在肿瘤血管生成方面起到关键的作用。靶向VEGF以及VEGFR的药物正在不断被开发出来，如bevacizumab、sorafenib、aflibercept等，这些药物以不同的方式抑制VEGF/VEGFR信号通路，在临床上已被证明能有效抑制多种肿瘤的生长，成功实现了相关药物的临床转化。研究认为，VEGF的信号转导途径很有可能是VEGFR-2通过活化PLC(磷脂酶C)，水解产生第二信使DAG(二酰甘油)，进而激活PKC(蛋白激酶)，PKC再激活RAF，RAF通过活化MEK最终激活MAPK。表明了BRAF和VEGFR-2两个靶点在肿瘤的发生发展过程中具有一定的协同作用。The "sprouting angiogenesis" mediated by vascular endothelial growth factor (VEGF) and its receptor VEGFR (VEGF receptor) plays an important role in both physiological and pathological angiogenesis. role, especially in tumor angiogenesis. Drugs targeting VEGF and VEGFR are being continuously developed, such as bevacizumab, sorafenib, aflibercept, etc. These drugs inhibit the VEGF/VEGFR signaling pathway in different ways, and have been clinically proven to be effective in inhibiting the growth of various tumors. The clinical transformation of related drugs has been realized. Studies suggest that the signal transduction pathway of VEGF is likely to be that VEGFR-2 activates PLC (phospholipase C), hydrolyzes to produce the second messenger DAG (diacylglycerol), and then activates PKC (protein kinase), and PKC activates RAF again. RAF ultimately activates MAPK by activating MEK. It shows that the two targets of BRAF and VEGFR-2 have a certain synergistic effect in the occurrence and development of tumors.

国内外研究现状及分析：Sorafenib是第1个报道的ⅡA型BRAF激酶抑制剂。但是在转移性黑色素瘤的Ⅲ期临床试验中，Sorafenib并没有显著疗效。一种可能是，黑色素瘤依赖周围的新生血管进行转移，另一种可能是，在黑色素瘤中，Sorafenib对RAF激酶没有特异性的作用。而2011年BRAF^V600E抑制剂Vemurafenib的上市成为一个重要突破。约有一半的晚期黑色素瘤有变异，在这个人群中Vemurafenib的应答率达到50％，比化疗延长3个月生存期。这个产品当时被认为是黑色素瘤治疗的重要进展，也是个体化治疗的一个成功典范。但不幸的是，在半年左右开始出现耐药。而另一种RAF抑制剂RAF265也被报道为RAF/VEGFR激酶双靶点抑制剂，抑制VEGF诱导的血管生成和突变BRAF，作用于A375M(BRAF^V600E)人黑色素瘤细胞系，能够有效的降低肿瘤糖代谢和FDG累积。因此，以BRAF^V600E和VEGFR-2为靶点，设计合成BRAF^V600E/VEGFR-2激酶的双靶点抑制剂，通过使BRAF^V600E和VEGFR-2表达减少，阻断其信号转导通路或耗竭肿瘤细胞产生的血管内皮生长因子而抑制肿瘤血管的生成，切断肿瘤血供，则有望达到抑制肿瘤生长、发展和转移的目的。而双靶点抑制剂在抗肿瘤方面具有更高的选择性和更好的活性。Research status and analysis at home and abroad: Sorafenib is the first reported type ⅡA BRAF kinase inhibitor. However, in phase III clinical trials of metastatic melanoma, Sorafenib did not have a significant effect. One possibility is that melanoma relies on surrounding neovascularization for metastasis, and the other possibility is that Sorafenib has no specific effect on RAF kinase in melanoma. In 2011, the launch of the BRAF^V600E inhibitor Vemurafenib became an important breakthrough. About half of advanced melanomas have mutations, and the response rate of Vemurafenib in this population reaches 50%, prolonging survival by 3 months compared with chemotherapy. This product was considered an important advance in the treatment of melanoma and a successful example of individualized treatment. But unfortunately, drug resistance began to appear in about half a year. Another RAF inhibitor, RAF265, has also been reported as a RAF/VEGFR kinase dual-target inhibitor, inhibiting VEGF-induced angiogenesis and mutant BRAF, and acting on the A375M (BRAF^V600E ) human melanoma cell line, which can effectively reduce tumor Glucose metabolism and FDG accumulation. Therefore, targeting BRAF^V600E and VEGFR-2, design and synthesize a dual-target inhibitor of BRAF^V600E /VEGFR-2 kinase, which can block the signal transduction pathway or deplete tumors by reducing the expression of BRAF^V600E and VEGFR-2 The vascular endothelial growth factor produced by the cells inhibits the formation of tumor angiogenesis and cuts off the tumor blood supply, which is expected to achieve the purpose of inhibiting tumor growth, development and metastasis. The dual-target inhibitors have higher selectivity and better activity in anti-tumor.

因此，本领域迫切需要开发出结构新颖，活性强的BRAF/VEGFR-2激酶的双靶点抑制剂。Therefore, there is an urgent need in this field to develop dual-target inhibitors of BRAF/VEGFR-2 kinase with novel structure and strong activity.

发明内容Contents of the invention

本发明的目的在于提供一种具有抗肿瘤活性的1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶类化合物和其衍生物、以及该物质的合成方法和应用。The object of the present invention is to provide a kind of 1-(2-tetrahydropyran)-1H-pyrazolo[3,4-d]pyrimidine compound and its derivatives with anti-tumor activity, and the synthetic method of the substance and apply.

本发明的技术方案如下：Technical scheme of the present invention is as follows:

一种具有抗肿瘤活性的1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶类化合物，其通式(Ⅰ)如下：A 1-(2-tetrahydropyran)-1H-pyrazolo[3,4-d]pyrimidine compound with antitumor activity, its general formula (I) is as follows:

其中X为氧，硫或氮；Wherein X is oxygen, sulfur or nitrogen;

连接基团Linker为取代在母体苯环的2位、3位或4位的其中N原子一端与母体相连；The linking group Linker is substituted at the 2-position, 3-position or 4-position of the parent benzene ring One end of the N atom is connected to the parent body;

R₁为-H、卤素、C₁～C₄烷基、C₁～C₄烷氧基、C₁～C₄烷硫基或-NO₂；R₂为取代的4～12元芳杂环基、所述取代芳杂环上的取代基为-H、C₁～C₆烷基、芳基、-CF₃或5～10元芳杂环基；所述芳杂环基上的杂原子个数为1～4个，杂原子为O、S或N；R₃为-H、卤素、烷基、-CF₃、-NO₂或-CN；R₄为-H或烷基。R₁ is -H, halogen, C₁ to C₄ alkyl, C₁ to C₄ alkoxy, C₁ to C₄ alkylthio or -NO₂ ; R₂ is a substituted 4 to 12 membered aromatic heterocycle base, The substituent on the substituted aromatic heterocycle is -H, C₁ ~C₆ alkyl, aryl, -CF₃ or 5-10 membered aromatic heterocycle; the number of heteroatoms on the aromatic heterocycle is is 1 to 4, and the heteroatom is O, S or N; R₃ is -H, halogen, alkyl, -CF₃ , -NO₂ or -CN; R₄ is -H or alkyl.

所述的一种具有抗肿瘤活性的1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶类化合物，通式(Ⅰ)中X为氧；连接基团Linker为取代在母体苯环的2位、3位或4位的其中N原子一端与母体相连；R₁为-H、卤素、C₁～C₄烷基、C₁～C₄烷氧基、C₁～C₄烷硫基或-NO₂；R₂为取代的4～12元芳杂环基、所述取代芳杂环上的取代基为-H、C₁～C₆烷基、芳基、-CF₃或5～10元芳杂环基；所述芳杂环基上的杂原子个数为1～4个，杂原子为O或S；R₃为-H、卤素、烷基、-CF₃、-NO₂或-CN；R₄为-H；R₃、R₄所含的氢原子各自独立地被一个或多个相同或不同的G¹或G²取代；G¹或G²各自独立地选自H、-OH、-NH₂、-CN、-CF₃、卤素、C_1-6烷基、C_3-6环烷基、C_2-6烯基、C_2-6炔基、C₆芳基、5-6元杂环芳基、C_3-6杂脂环基、C_1-6烷氧基、C_3-6环烷氧基、C₆芳氧基、5-7元杂芳氧基、C_3-6杂脂环氧基、C_1-6烷氨基、C_3-6环烷氨基、C₆芳氨基、5-7元杂环芳氨基、C_3-6杂脂环氨基、C_1-6烷氧基-CO-、C_3-6环烷氧基-CO-、C₆芳氧基-CO-、5-7元杂环芳氧基-CO-、C_3-6杂脂环氧基-CO-、C_1-6烷氨基-CO-、C_3-6环烷氨基-CO-、C₆芳氨基-CO-、5-7元杂环芳氨基-CO-、C_3-6杂脂环氨基-CO-中的其中一种。The 1-(2-tetrahydropyran)-1H-pyrazolo[3,4-d]pyrimidine compound with anti-tumor activity, X in the general formula (I) is oxygen; the linking group Linker is substituted at the 2-position, 3-position or 4-position of the parent benzene ring One end of the N atom is connected to the parent; R₁ is -H, halogen, C₁ to C₄ alkyl, C₁ to C₄ alkoxy, C₁ to C₄ alkylthio or -NO₂ ; R₂ is substituted 4-12 membered aromatic heterocyclic groups, The substituent on the substituted aromatic heterocycle is -H, C₁ ~C₆ alkyl, aryl, -CF₃ or 5-10 membered aromatic heterocycle; the number of heteroatoms on the aromatic heterocycle is 1 to 4, heteroatoms are O or S; R₃ is -H, halogen, alkyl, -CF₃ , -NO₂ or -CN; R₄ is -H; R₃ and R₄ contain hydrogen Atoms are each independently substituted by one or more identical or different G¹ or G² ; each G¹ or G² is independently selected from H, -OH, -NH₂ , -CN, -CF₃ , halogen, C_{1 -6} alkyl, C_3-6 cycloalkyl, C_2-6 alkenyl, C_2-6 alkynyl, C₆ aryl, 5-6 membered heterocyclic aryl, C_3-6 heteroalicyclic, C_1-6 alkoxy, C_3-6 cycloalkoxy, C₆ aryloxy, 5-7 heteroaryloxy, C_3-6 heteroaliphatic epoxy, C_1-6 alkylamino, C_3-6 cycloalkylamino, C₆ arylamino, 5-7 membered heterocyclic arylamino, C_3-6 heteroalicyclic amino, C_1-6 alkoxy-CO-, C_3-6 cycloalkoxy- CO-, C₆ aryloxy-CO-, 5-7 membered heterocyclic aryloxy-CO-, C_3-6 heteroaliphatic epoxy-CO-, C_1-6 alkylamino-CO-, C₃ One of_-6 cycloalkylamino-CO-, C₆ arylamino-CO-, 5-7 membered heterocyclic arylamino-CO-, C_3-6 heteroalicyclic amino-CO-.

通式(Ⅰ)所示的化合物或其在药学上可接受的盐、水合物、溶剂合物、多晶型物、互变异构体或前药，该化合物选自1-(3-甲基苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲、1-(3,4-二氯苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲、1-(2-氯-5-甲基苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲、1-(2,3-二甲基苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲、1-(4-氟-3-甲基苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲、1-(3-硝基苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲、1-(2-氟-5-(三氟甲基)苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲、1-(3-氰基苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲、1-(5-甲基-2-硝基苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲、1-(2-甲基苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲、1-(3-氟苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲、1-(4-氟苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲、1-(2,5-二甲基苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲、1-(3-三氟甲基苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲、1-(3-三氟甲基-4-氯苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲、1-(3-三氟-5-三氟甲基苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲、1-(4-氯苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲、1-(2-氯-3-三氟甲基苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲、1-环己基-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲、1-(3-甲基苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]硫脲、1-(4-氯苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]硫脲、1-(3-三氟甲基-4-氯苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]硫脲、1-(3-三氟甲基苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]硫脲中的其中一种。The compound represented by the general formula (I) or its pharmaceutically acceptable salt, hydrate, solvate, polymorph, tautomer or prodrug, the compound is selected from the group consisting of 1-(3-methyl phenyl)-3-[4-[[1-(2-tetrahydropyran)-1H-pyrazolo[3,4-d]pyrimidine]-4-oxyl]phenyl]urea, 1- (3,4-Dichlorophenyl)-3-[4-[[1-(2-tetrahydropyran)-1H-pyrazolo[3,4-d]pyrimidine]-4-oxyl]benzene base]urea, 1-(2-chloro-5-methylphenyl)-3-[4-[[1-(2-tetrahydropyran)-1H-pyrazolo[3,4-d]pyrimidine ]-4-oxyl]phenyl]urea, 1-(2,3-dimethylphenyl)-3-[4-[[1-(2-tetrahydropyran)-1H-pyrazolo[ 3,4-d]pyrimidine]-4-oxyl]phenyl]urea, 1-(4-fluoro-3-methylphenyl)-3-[4-[[1-(2-tetrahydropyran )-1H-pyrazolo[3,4-d]pyrimidine]-4-oxyl]phenyl]urea, 1-(3-nitrophenyl)-3-[4-[[1-(2- Tetrahydropyran)-1H-pyrazolo[3,4-d]pyrimidine]-4-oxyl]phenyl]urea, 1-(2-fluoro-5-(trifluoromethyl)phenyl)- 3-[4-[[1-(2-tetrahydropyran)-1H-pyrazolo[3,4-d]pyrimidine]-4-oxyl]phenyl]urea, 1-(3-cyano Phenyl)-3-[4-[[1-(2-tetrahydropyran)-1H-pyrazolo[3,4-d]pyrimidine]-4-oxyl]phenyl]urea, 1-( 5-methyl-2-nitrophenyl)-3-[4-[[1-(2-tetrahydropyran)-1H-pyrazolo[3,4-d]pyrimidine]-4-oxyl ]phenyl]urea, 1-(2-methylphenyl)-3-[4-[[1-(2-tetrahydropyran)-1H-pyrazolo[3,4-d]pyrimidine]- 4-Oxy]phenyl]urea, 1-(3-fluorophenyl)-3-[4-[[1-(2-tetrahydropyran)-1H-pyrazolo[3,4-d] Pyrimidine]-4-oxyl]phenyl]urea, 1-(4-fluorophenyl)-3-[4-[[1-(2-tetrahydropyran)-1H-pyrazolo[3,4 -d]pyrimidine]-4-oxyl]phenyl]urea, 1-(2,5-dimethylphenyl)-3-[4-[[1-(2-tetrahydropyran)-1H- Pyrazolo[3,4-d]pyrimidine]-4-oxyl]phenyl]urea, 1-(3-trifluoromethylphenyl)-3-[4-[[1-(2-tetrahydro Pyran)-1H-pyrazolo[3,4-d]pyrimidine]-4-oxyl]phenyl]urea, 1-(3-trifluoromethyl-4-chlorophenyl)-3-[4 -[[1-(2-tetrahydropyran)-1H-pyrazolo[3,4-d]pyrimidine]-4-oxyl]phenyl]urea, 1-(3-trifluoro-5-tri Fluoromethylphenyl)-3-[4-[[1-(2-tetrahydropyran)-1H-pyrazolo[3,4-d]pyrimidine]-4-oxyl]phenyl]urea, 1-(4-Chlorophenyl)- 3-[4-[[1-(2-tetrahydropyran)-1H-pyrazolo[3,4-d]pyrimidine]-4-oxyl]phenyl]urea, 1-(2-chloro- 3-trifluoromethylphenyl)-3-[4-[[1-(2-tetrahydropyran)-1H-pyrazolo[3,4-d]pyrimidine]-4-oxyl]phenyl ]urea, 1-cyclohexyl-3-[4-[[1-(2-tetrahydropyran)-1H-pyrazolo[3,4-d]pyrimidine]-4-oxyl]phenyl]urea , 1-(3-methylphenyl)-3-[4-[[1-(2-tetrahydropyran)-1H-pyrazolo[3,4-d]pyrimidine]-4-oxyl] Phenyl]thiourea, 1-(4-chlorophenyl)-3-[4-[[1-(2-tetrahydropyran)-1H-pyrazolo[3,4-d]pyrimidine]-4 -Oxy]phenyl]thiourea, 1-(3-trifluoromethyl-4-chlorophenyl)-3-[4-[[1-(2-tetrahydropyran)-1H-pyrazolo [3,4-d]pyrimidine]-4-oxyl]phenyl]thiourea, 1-(3-trifluoromethylphenyl)-3-[4-[[1-(2-tetrahydropyran )-1H-pyrazolo[3,4-d]pyrimidine]-4-oxyl]phenyl]thiourea.

所述的一种具有抗肿瘤活性的1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶类化合物合成方法：将4,6-二氯-5-嘧啶甲醛与一水合肼进行环合反应生成4-氯-1H-吡唑并[3,4-d]嘧啶，在催化剂作用下，与3,4-二氢-2H-吡喃反应，生成4-氯-1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶，在保护气体保护下，与对氨基酚反应得到胺类中间体4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯胺，最后与异氰酸酯或硫异氰酸酯等反应得到目标化合物。The synthetic method of the 1-(2-tetrahydropyran)-1H-pyrazolo[3,4-d]pyrimidine compound with anti-tumor activity: 4,6-dichloro-5-pyrimidine Formaldehyde and hydrazine monohydrate undergo ring closure reaction to generate 4-chloro-1H-pyrazolo[3,4-d]pyrimidine, under the action of catalyst, react with 3,4-dihydro-2H-pyran to generate 4- Chloro-1-(2-tetrahydropyran)-1H-pyrazolo[3,4-d]pyrimidine, under the protection of protective gas, reacts with p-aminophenol to obtain amine intermediate 4-[[1-( 2-tetrahydropyran)-1H-pyrazolo[3,4-d]pyrimidine]-4-oxyl]aniline, and finally react with isocyanate or thioisocyanate to obtain the target compound.

所述的催化剂为酸性催化剂和盐类催化剂中的至少一种。The catalyst is at least one of an acid catalyst and a salt catalyst.

所述的保护气体为氮气和惰性气体中的至少一种。The protective gas is at least one of nitrogen and inert gas.

一种药物组合物，包括下列成分中的至少一种：a)化合物、b)该化合物在药学上可接受的盐、c)该化合物的水合物、d)该化合物的溶剂合物、e)该化合物的多晶型物、f)该化合物的互变异构体、g)该化合物的前药；其中，所述的化合物为通式(Ⅰ)所示的化合物。A pharmaceutical composition comprising at least one of the following components: a) a compound, b) a pharmaceutically acceptable salt of the compound, c) a hydrate of the compound, d) a solvate of the compound, e) A polymorph of the compound, f) a tautomer of the compound, g) a prodrug of the compound; wherein, the compound is a compound represented by general formula (I).

所述的一种药物组合物在治疗因蛋白激酶活性异常所引起疾病的应用。The application of the pharmaceutical composition in treating diseases caused by abnormal protein kinase activity.

所述的疾病为癌症。The disease is cancer.

所述的疾病为黑色素瘤、肝癌、肾癌、肺癌、骨癌、胰腺癌、皮肤癌、头颈癌、皮肤或眼内黑素瘤、子宫癌、卵巢癌、直肠癌、肛门区癌、胃癌、结肠癌、乳腺癌、输卵管癌、子宫内膜癌、宫颈癌、阴道癌、阴户癌、何杰金病、食道癌、小肠癌、内分泌系统癌、甲状腺癌、甲状旁腺癌、软组织肉瘤、尿道癌、阴茎癌、前列腺癌、慢性或急性白血病、膀胱癌、肾或输尿管癌、脊柱轴肿瘤、垂体腺瘤、胃肠间质肿瘤、结肠直肠癌、非小细胞肺癌、小细胞肺癌、肥大细胞增多症、胶质瘤、肉瘤、淋巴瘤中的一种或任意几种的组合。The diseases mentioned are melanoma, liver cancer, kidney cancer, lung cancer, bone cancer, pancreatic cancer, skin cancer, head and neck cancer, skin or intraocular melanoma, uterine cancer, ovarian cancer, rectal cancer, anal region cancer, gastric cancer, Colon cancer, breast cancer, fallopian tube cancer, endometrial cancer, cervical cancer, vaginal cancer, vulvar cancer, Hodgkin's disease, esophagus cancer, small intestine cancer, endocrine system cancer, thyroid cancer, parathyroid cancer, soft tissue sarcoma, urethra Carcinoma, penile cancer, prostate cancer, chronic or acute leukemia, bladder cancer, kidney or ureter cancer, spinal axis tumors, pituitary adenomas, gastrointestinal stromal tumors, colorectal cancer, non-small cell lung cancer, small cell lung cancer, mast cell One or any combination of polycythemia, glioma, sarcoma, and lymphoma.

本发明的有益效果是：The beneficial effects of the present invention are:

本发明开发出一种结构新颖，具有显著抗肿瘤活性的吡唑并[3,4-d]嘧啶类化合物，该化合物可以用作为BRAF/VEGFR-2激酶的双靶点抑制剂，在治疗因蛋白激酶活性异常所引起疾病方面有良好的效果和广阔的应用前景。The present invention develops a pyrazolo[3,4-d]pyrimidine compound with novel structure and significant antitumor activity, which can be used as a dual-target inhibitor of BRAF/VEGFR-2 kinase, and is used in the treatment of It has good effect and broad application prospect in diseases caused by abnormal protein kinase activity.

具体实施方式Detailed ways

一种具有抗肿瘤活性的1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶类化合物，其通式(Ⅰ)如下：A 1-(2-tetrahydropyran)-1H-pyrazolo[3,4-d]pyrimidine compound with antitumor activity, its general formula (I) is as follows:

其中X为氧，硫或氮；Wherein X is oxygen, sulfur or nitrogen;

连接基团Linker为取代在母体苯环的2位、3位或4位的其中N原子一端与母体相连；The linking group Linker is substituted at the 2-position, 3-position or 4-position of the parent benzene ring One end of the N atom is connected to the parent body;

R₁为-H、卤素、C₁～C₄烷基、C₁～C₄烷氧基、C₁～C₄烷硫基或-NO₂；R₂为取代的4～12元芳杂环基、所述取代芳杂环上的取代基为-H、C₁～C₆烷基、芳基、-CF₃或5～10元芳杂环基；所述芳杂环基上的杂原子个数为1～4个，杂原子为O、S或N；R₃为-H、卤素、烷基、-CF₃、-NO₂或-CN；R₄为-H或烷基。R₁ is -H, halogen, C₁ to C₄ alkyl, C₁ to C₄ alkoxy, C₁ to C₄ alkylthio or -NO₂ ; R₂ is a substituted 4 to 12 membered aromatic heterocycle base, The substituent on the substituted aromatic heterocycle is -H, C₁ ~C₆ alkyl, aryl, -CF₃ or 5-10 membered aromatic heterocycle; the number of heteroatoms on the aromatic heterocycle is is 1 to 4, and the heteroatom is O, S or N; R₃ is -H, halogen, alkyl, -CF₃ , -NO₂ or -CN; R₄ is -H or alkyl.

所述的一种具有抗肿瘤活性的1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶类化合物，通式(Ⅰ)中X为氧；连接基团Linker为取代在母体苯环的2位、3位或4位的其中N原子一端与母体相连；R₁为-H、卤素、C₁～C₄烷基、C₁～C₄烷氧基、C₁～C₄烷硫基或-NO₂；R₂为取代的4～12元芳杂环基、所述取代芳杂环上的取代基为-H、C₁～C₆烷基、芳基、-CF₃或5～10元芳杂环基；所述芳杂环基上的杂原子个数为1～4个，杂原子为O或S；R₃为-H、卤素、烷基、-CF₃、-NO₂或-CN；R₄为-H；R₃、R₄所含的氢原子各自独立地被一个或多个相同或不同的G¹或G²取代；G¹或G²各自独立地选自H、-OH、-NH₂、-CN、-CF₃、卤素、C_1-6烷基、C_3-6环烷基、C_2-6烯基、C_2-6炔基、C₆芳基、5-6元杂环芳基、C_3-6杂脂环基、C_1-6烷氧基、C_3-6环烷氧基、C₆芳氧基、5-7元杂芳氧基、C_3-6杂脂环氧基、C_1-6烷氨基、C_3-6环烷氨基、C₆芳氨基、5-7元杂环芳氨基、C_3-6杂脂环氨基、C_1-6烷氧基-CO-、C_3-6环烷氧基-CO-、C₆芳氧基-CO-、5-7元杂环芳氧基-CO-、C_3-6杂脂环氧基-CO-、C_1-6烷氨基-CO-、C_3-6环烷氨基-CO-、C₆芳氨基-CO-、5-7元杂环芳氨基-CO-、C_3-6杂脂环氨基-CO-中的其中一种。The 1-(2-tetrahydropyran)-1H-pyrazolo[3,4-d]pyrimidine compound with anti-tumor activity, X in the general formula (I) is oxygen; the linking group Linker is substituted at the 2-position, 3-position or 4-position of the parent benzene ring One end of the N atom is connected to the parent; R₁ is -H, halogen, C₁ to C₄ alkyl, C₁ to C₄ alkoxy, C₁ to C₄ alkylthio or -NO₂ ; R₂ is substituted 4-12 membered aromatic heterocyclic groups, The substituent on the substituted aromatic heterocycle is -H, C₁ ~C₆ alkyl, aryl, -CF₃ or 5-10 membered aromatic heterocycle; the number of heteroatoms on the aromatic heterocycle is 1 to 4, heteroatoms are O or S; R₃ is -H, halogen, alkyl, -CF₃ , -NO₂ or -CN; R₄ is -H; R₃ and R₄ contain hydrogen Atoms are each independently substituted by one or more identical or different G¹ or G² ; each G¹ or G² is independently selected from H, -OH, -NH₂ , -CN, -CF₃ , halogen, C_{1 -6} alkyl, C_3-6 cycloalkyl, C_2-6 alkenyl, C_2-6 alkynyl, C₆ aryl, 5-6 membered heterocyclic aryl, C_3-6 heteroalicyclic, C_1-6 alkoxy, C_3-6 cycloalkoxy, C₆ aryloxy, 5-7 heteroaryloxy, C_3-6 heteroaliphatic epoxy, C_1-6 alkylamino, C_3-6 cycloalkylamino, C₆ arylamino, 5-7 membered heterocyclic arylamino, C_3-6 heteroalicyclic amino, C_1-6 alkoxy-CO-, C_3-6 cycloalkoxy- CO-, C₆ aryloxy-CO-, 5-7 membered heterocyclic aryloxy-CO-, C_3-6 heteroaliphatic epoxy-CO-, C_1-6 alkylamino-CO-, C₃ One of_-6 cycloalkylamino-CO-, C₆ arylamino-CO-, 5-7 membered heterocyclic arylamino-CO-, C_3-6 heteroalicyclic amino-CO-.

通式(Ⅰ)所示的化合物或其在药学上可接受的盐、水合物、溶剂合物、多晶型物、互变异构体或前药，该化合物选自1-(3-甲基苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲、1-(3,4-二氯苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲、1-(2-氯-5-甲基苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲、1-(2,3-二甲基苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲、1-(4-氟-3-甲基苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲、1-(3-硝基苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲、1-(2-氟-5-(三氟甲基)苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲、1-(3-氰基苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲、1-(5-甲基-2-硝基苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲、1-(2-甲基苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲、1-(3-氟苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲、1-(4-氟苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲、1-(2,5-二甲基苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲、1-(3-三氟甲基苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲、1-(3-三氟甲基-4-氯苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲、1-(3-三氟-5-三氟甲基苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲、1-(4-氯苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲、1-(2-氯-3-三氟甲基苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲、1-环己基-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲、1-(3-甲基苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]硫脲、1-(4-氯苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]硫脲、1-(3-三氟甲基-4-氯苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]硫脲、1-(3-三氟甲基苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]硫脲中的其中一种。The compound represented by the general formula (I) or its pharmaceutically acceptable salt, hydrate, solvate, polymorph, tautomer or prodrug, the compound is selected from the group consisting of 1-(3-methyl phenyl)-3-[4-[[1-(2-tetrahydropyran)-1H-pyrazolo[3,4-d]pyrimidine]-4-oxyl]phenyl]urea, 1- (3,4-Dichlorophenyl)-3-[4-[[1-(2-tetrahydropyran)-1H-pyrazolo[3,4-d]pyrimidine]-4-oxyl]benzene base]urea, 1-(2-chloro-5-methylphenyl)-3-[4-[[1-(2-tetrahydropyran)-1H-pyrazolo[3,4-d]pyrimidine ]-4-oxyl]phenyl]urea, 1-(2,3-dimethylphenyl)-3-[4-[[1-(2-tetrahydropyran)-1H-pyrazolo[ 3,4-d]pyrimidine]-4-oxyl]phenyl]urea, 1-(4-fluoro-3-methylphenyl)-3-[4-[[1-(2-tetrahydropyran )-1H-pyrazolo[3,4-d]pyrimidine]-4-oxyl]phenyl]urea, 1-(3-nitrophenyl)-3-[4-[[1-(2- Tetrahydropyran)-1H-pyrazolo[3,4-d]pyrimidine]-4-oxyl]phenyl]urea, 1-(2-fluoro-5-(trifluoromethyl)phenyl)- 3-[4-[[1-(2-tetrahydropyran)-1H-pyrazolo[3,4-d]pyrimidine]-4-oxyl]phenyl]urea, 1-(3-cyano Phenyl)-3-[4-[[1-(2-tetrahydropyran)-1H-pyrazolo[3,4-d]pyrimidine]-4-oxyl]phenyl]urea, 1-( 5-methyl-2-nitrophenyl)-3-[4-[[1-(2-tetrahydropyran)-1H-pyrazolo[3,4-d]pyrimidine]-4-oxyl ]phenyl]urea, 1-(2-methylphenyl)-3-[4-[[1-(2-tetrahydropyran)-1H-pyrazolo[3,4-d]pyrimidine]- 4-Oxy]phenyl]urea, 1-(3-fluorophenyl)-3-[4-[[1-(2-tetrahydropyran)-1H-pyrazolo[3,4-d] Pyrimidine]-4-oxyl]phenyl]urea, 1-(4-fluorophenyl)-3-[4-[[1-(2-tetrahydropyran)-1H-pyrazolo[3,4 -d]pyrimidine]-4-oxyl]phenyl]urea, 1-(2,5-dimethylphenyl)-3-[4-[[1-(2-tetrahydropyran)-1H- Pyrazolo[3,4-d]pyrimidine]-4-oxyl]phenyl]urea, 1-(3-trifluoromethylphenyl)-3-[4-[[1-(2-tetrahydro Pyran)-1H-pyrazolo[3,4-d]pyrimidine]-4-oxyl]phenyl]urea, 1-(3-trifluoromethyl-4-chlorophenyl)-3-[4 -[[1-(2-tetrahydropyran)-1H-pyrazolo[3,4-d]pyrimidine]-4-oxyl]phenyl]urea, 1-(3-trifluoro-5-tri Fluoromethylphenyl)-3-[4-[[1-(2-tetrahydropyran)-1H-pyrazolo[3,4-d]pyrimidine]-4-oxyl]phenyl]urea, 1-(4-Chlorophenyl)- 3-[4-[[1-(2-tetrahydropyran)-1H-pyrazolo[3,4-d]pyrimidine]-4-oxyl]phenyl]urea, 1-(2-chloro- 3-trifluoromethylphenyl)-3-[4-[[1-(2-tetrahydropyran)-1H-pyrazolo[3,4-d]pyrimidine]-4-oxyl]phenyl ]urea, 1-cyclohexyl-3-[4-[[1-(2-tetrahydropyran)-1H-pyrazolo[3,4-d]pyrimidine]-4-oxyl]phenyl]urea , 1-(3-methylphenyl)-3-[4-[[1-(2-tetrahydropyran)-1H-pyrazolo[3,4-d]pyrimidine]-4-oxyl] Phenyl]thiourea, 1-(4-chlorophenyl)-3-[4-[[1-(2-tetrahydropyran)-1H-pyrazolo[3,4-d]pyrimidine]-4 -Oxy]phenyl]thiourea, 1-(3-trifluoromethyl-4-chlorophenyl)-3-[4-[[1-(2-tetrahydropyran)-1H-pyrazolo [3,4-d]pyrimidine]-4-oxyl]phenyl]thiourea, 1-(3-trifluoromethylphenyl)-3-[4-[[1-(2-tetrahydropyran )-1H-pyrazolo[3,4-d]pyrimidine]-4-oxyl]phenyl]thiourea.

所述的一种具有抗肿瘤活性的1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶类化合物合成方法：将4,6-二氯-5-嘧啶甲醛与一水合肼进行环合反应生成4-氯-1H-吡唑并[3,4-d]嘧啶，在催化剂作用下，与3,4-二氢-2H-吡喃反应，生成4-氯-1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶，在保护气体保护下，与对氨基酚反应得到胺类中间体4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯胺，最后与异氰酸酯或硫异氰酸酯等反应得到目标化合物。具体合成线路如下：The synthetic method of the 1-(2-tetrahydropyran)-1H-pyrazolo[3,4-d]pyrimidine compound with anti-tumor activity: 4,6-dichloro-5-pyrimidine Formaldehyde and hydrazine monohydrate undergo ring closure reaction to generate 4-chloro-1H-pyrazolo[3,4-d]pyrimidine, under the action of catalyst, react with 3,4-dihydro-2H-pyran to generate 4- Chloro-1-(2-tetrahydropyran)-1H-pyrazolo[3,4-d]pyrimidine, under the protection of protective gas, reacts with p-aminophenol to obtain amine intermediate 4-[[1-( 2-tetrahydropyran)-1H-pyrazolo[3,4-d]pyrimidine]-4-oxyl]aniline, and finally react with isocyanate or thioisocyanate to obtain the target compound. The specific synthetic route is as follows:

式中a：Et₃N,MeOH；b:PPTs,EtOAc；c:取代的苯酚,Cs₂CO₃,DMF,N₂；d:异氰酸或硫异氰酸酯,CH₂Cl₂。In the formula, a: Et₃ N, MeOH; b: PPTs, EtOAc; c: substituted phenol, Cs₂ CO₃ , DMF, N₂ ; d: isocyanic acid or thioisocyanate, CH₂ Cl₂ .

合成式中最后的产物即为通式(Ⅰ)所示的化合物。The final product in the synthetic formula is the compound shown in general formula (I).

优选的，所述的催化剂为酸性催化剂和盐类催化剂中的至少一种；进一步优选的，所述的催化剂为盐类催化剂；再进一步优选的，所述的催化剂为对甲苯磺酸吡啶盐。Preferably, the catalyst is at least one of an acid catalyst and a salt catalyst; more preferably, the catalyst is a salt catalyst; still more preferably, the catalyst is pyridinium p-toluenesulfonate.

优选的，所述的保护气体为氮气和惰性气体中的至少一种；进一步优选的，所述的保护气体为氮气、氩气中的其中一种；再进一步优选的，所述的保护气体为氮气。Preferably, the protective gas is at least one of nitrogen and an inert gas; more preferably, the protective gas is one of nitrogen and argon; still more preferably, the protective gas is nitrogen.

一种药物组合物，包括下列成分中的至少一种：a)化合物、b)该化合物在药学上可接受的盐、c)该化合物的水合物、d)该化合物的溶剂合物、e)该化合物的多晶型物、f)该化合物的互变异构体、g)该化合物的前药；其中，所述的化合物为通式(Ⅰ)所示的化合物。A pharmaceutical composition comprising at least one of the following components: a) a compound, b) a pharmaceutically acceptable salt of the compound, c) a hydrate of the compound, d) a solvate of the compound, e) A polymorph of the compound, f) a tautomer of the compound, g) a prodrug of the compound; wherein, the compound is a compound represented by general formula (I).

进一步优选的，所述的药物组合物还包括辅料；再进一步优选的，所述的辅料包括下列物质中的至少一种：溶剂、抛射剂、增溶剂、稳定剂、助流剂、矫味剂、防腐剂、助悬剂、包衣材料、芳香剂、抗黏合剂、整合剂、渗透促进剂、pH值调节剂、缓冲剂、增塑剂、助溶剂、乳化剂、着色剂、黏合剂、崩解剂、填充剂、润滑剂、润湿剂、渗透压调节剂、表面活性剂、发泡剂、消泡剂、增稠剂、包合剂、保湿剂、吸收剂、稀释剂、絮凝剂与反絮凝剂、助滤剂、释放阻滞剂。Further preferably, the pharmaceutical composition also includes adjuvants; still further preferably, the adjuvants include at least one of the following substances: solvents, propellants, solubilizers, stabilizers, glidants, flavoring agents , preservatives, suspending agents, coating materials, fragrances, anti-adhesives, integration agents, penetration enhancers, pH regulators, buffers, plasticizers, solubilizers, emulsifiers, colorants, binders, Disintegrants, fillers, lubricants, wetting agents, osmotic pressure regulators, surfactants, foaming agents, defoamers, thickeners, inclusion agents, humectants, absorbents, diluents, flocculants and Deflocculants, filter aids, release retardants.

本发明的药物组合物可制成各种剂型：Pharmaceutical composition of the present invention can be made into various dosage forms:

按照剂型的分散系统进行分类，具体来说，可以制成以下剂型：溶液型、胶体溶液型、乳剂型、混悬型、气体分散型、微粒分散型、固体分散型；Classified according to the dispersion system of the dosage form, specifically, it can be made into the following dosage forms: solution type, colloid solution type, emulsion type, suspension type, gas dispersion type, particle dispersion type, solid dispersion type;

按照形态分类，具体来说，可以制成以下剂型：液体剂型(如芳香水剂、溶液剂、注射剂、合剂、洗剂、搽剂等)，气体剂型(如气雾剂、喷雾剂等)，固体剂型(如散剂、丸剂、片剂、膜剂等)，半固体剂型(如软膏剂、栓剂、糊剂等)；Classified according to the form, specifically, it can be made into the following dosage forms: liquid dosage forms (such as aromatic water, solution, injection, mixture, lotion, liniment, etc.), gas dosage forms (such as aerosol, spray, etc.), Solid dosage forms (such as powders, pills, tablets, films, etc.), semi-solid dosage forms (such as ointments, suppositories, pastes, etc.);

按照给药途径分类，具体来说，可以制成以下剂型：经胃肠道给药的剂型、不经胃肠道给药的剂型。Classified according to the route of administration, specifically, the following dosage forms can be prepared: a dosage form for gastrointestinal administration, and a dosage form for parenteral administration.

所述的一种药物组合物在治疗因蛋白激酶活性异常所引起疾病的应用。优选的，所述的疾病为癌症；进一步优选的，所述的疾病为黑色素瘤、肝癌、肾癌、肺癌、骨癌、胰腺癌、皮肤癌、头颈癌、皮肤或眼内黑素瘤、子宫癌、卵巢癌、直肠癌、肛门区癌、胃癌、结肠癌、乳腺癌、输卵管癌、子宫内膜癌、宫颈癌、阴道癌、阴户癌、何杰金病、食道癌、小肠癌、内分泌系统癌、甲状腺癌、甲状旁腺癌、软组织肉瘤、尿道癌、阴茎癌、前列腺癌、慢性或急性白血病、膀胱癌、肾或输尿管癌、脊柱轴肿瘤、垂体腺瘤、胃肠间质肿瘤、结肠直肠癌、非小细胞肺癌、小细胞肺癌、肥大细胞增多症、胶质瘤、肉瘤、淋巴瘤中的一种或任意几种的组合。The application of the pharmaceutical composition in treating diseases caused by abnormal protein kinase activity. Preferably, the disease is cancer; more preferably, the disease is melanoma, liver cancer, kidney cancer, lung cancer, bone cancer, pancreatic cancer, skin cancer, head and neck cancer, skin or intraocular melanoma, uterine Cancer, ovarian cancer, rectal cancer, anal region cancer, stomach cancer, colon cancer, breast cancer, fallopian tube cancer, endometrial cancer, cervical cancer, vaginal cancer, vulvar cancer, Hodgkin's disease, esophagus cancer, small bowel cancer, endocrine system Carcinoma, thyroid cancer, parathyroid cancer, soft tissue sarcoma, urethral cancer, penile cancer, prostate cancer, chronic or acute leukemia, bladder cancer, kidney or ureter cancer, spinal axis tumors, pituitary adenomas, gastrointestinal stromal tumors, colon One or any combination of rectal cancer, non-small cell lung cancer, small cell lung cancer, mastocytosis, glioma, sarcoma, and lymphoma.

优选的，通式(Ⅰ)所示的化合物或其在药学上可接受的盐、水合物、溶剂合物、多晶型物、互变异构体或前药在制备BRAF/VEGFR-2激酶抑制剂中的应用。Preferably, the compound represented by general formula (I) or its pharmaceutically acceptable salt, hydrate, solvate, polymorph, tautomer or prodrug is used in the preparation of BRAF/VEGFR-2 kinase application of inhibitors.

进一步优选的，通式(Ⅰ)所示的化合物或其在药学上可接受的盐、水合物、溶剂合物、多晶型物、互变异构体或前药在制备治疗和/或预防和/或延缓和/或辅助治疗和/或处理与BRAF/VEGFR-2激酶活性过高相关的疾病的药物中的应用；再进一步优选的，通式(Ⅰ)所示的化合物或其在药学上可接受的盐、水合物、溶剂合物、多晶型物、互变异构体或前药在制备抗肿瘤药物中的应用；更进一步优选的，下列化合物中的至少一种或其在药学上可接受的盐、水合物、溶剂合物、多晶型物、互变异构体或前药在制备抗肿瘤药物中的应用，如在制备抗前列腺癌药物中的应用，或制备抗黑色素瘤药物中的应用。Further preferably, the compound represented by general formula (I) or its pharmaceutically acceptable salt, hydrate, solvate, polymorph, tautomer or prodrug is used in the preparation of treatment and/or prophylaxis And/or delay and/or adjuvant treatment and/or application in medicines for treating diseases related to hyperactivity of BRAF/VEGFR-2 kinase; still further preferably, the compound shown in general formula (I) or its pharmaceutical The application of acceptable salts, hydrates, solvates, polymorphs, tautomers or prodrugs in the preparation of antitumor drugs; more preferably, at least one of the following compounds or its The use of pharmaceutically acceptable salts, hydrates, solvates, polymorphs, tautomers or prodrugs in the preparation of anti-tumor drugs, such as the application in the preparation of anti-prostate cancer drugs, or the preparation of anti-cancer drugs Use in melanoma drugs.

本发明中“药学上可接受的盐”是指把母体化合物中的碱性基团转换成盐的形式。药学上可接受的盐包括，但不仅限于，碱性基团例如胺(氨)基的无机或有机酸盐类。本发明药学上可接受的盐可以由母体化合物合成，即母体化合物中的碱性基团与1-4当量的酸在一个溶剂系统中反应。合适的盐列举在Remington’s Pharmaceutical Sciences,17thed.,Mack Publishing Company,Easton,Pa.,1985,1418和Journal of PharmaceuticalScience,66,2,1977中。"Pharmaceutically acceptable salt" in the present invention refers to the form in which the basic group in the parent compound is converted into a salt. Pharmaceutically acceptable salts include, but are not limited to, inorganic or organic acid salts of basic groups such as amine (amino) groups. The pharmaceutically acceptable salts of the present invention can be synthesized from the parent compound by reacting the basic group in the parent compound with 1-4 equivalents of acid in a solvent system. Suitable salts are listed in Remington's Pharmaceutical Sciences, 17thed., Mack Publishing Company, Easton, Pa., 1985, 1418 and Journal of Pharmaceutical Science, 66, 2, 1977.

药学上可接受的酸加成盐可以由无机和有机酸制备如下：由衍生酸加成盐的无机酸包括盐酸、氢溴酸、硫酸、硝酸、磷酸等。由衍生酸加成盐的有机酸包括乙酸、丙酸、乙醇酸、丙酮酸、草酸、苹果酸、丙二酸、琥珀酸、马来酸、富马酸、酒石酸、柠檬酸、苯甲酸、肉桂酸、扁桃酸、甲磺酸、乙磺酸、对甲苯磺酸、水杨酸、苯磺酸等。衍生酸加成盐的无机酸和有机酸尤其选自盐酸、磷酸、硫酸、硝酸、高氯酸、氢溴酸、乙酸、苯甲酸和对甲苯磺酸。Pharmaceutically acceptable acid addition salts can be prepared from inorganic and organic acids as follows: Inorganic acids from which acid addition salts are derived include hydrochloric, hydrobromic, sulfuric, nitric, phosphoric, and the like. Organic acids from which acid addition salts are derived include acetic, propionic, glycolic, pyruvic, oxalic, malic, malonic, succinic, maleic, fumaric, tartaric, citric, benzoic, cinnamon acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, benzenesulfonic acid, etc. Inorganic and organic acids from which acid addition salts are derived are especially selected from hydrochloric acid, phosphoric acid, sulfuric acid, nitric acid, perchloric acid, hydrobromic acid, acetic acid, benzoic acid and p-toluenesulfonic acid.

以下通过具体的实施例对本发明的内容作进一步详细的说明。The content of the present invention will be described in further detail below through specific examples.

实施例1：Example 1:

4-氯-1H-吡唑并[3,4-d]嘧啶的合成Synthesis of 4-chloro-1H-pyrazolo[3,4-d]pyrimidine

在100mL三口烧瓶中，加入1.0g 4,6-二氯-5-嘧啶甲醛，20mL甲醇，搅拌溶解，降温至-65℃，滴加0.97mL(1.2eq)的三乙胺。0.274mL(1.0eq)一水合肼用10mL甲醇稀释，用滴液漏斗缓慢滴下。滴毕恢复至室温，反应2～3h，TLC监测。反应结束，旋蒸干溶剂，干燥。用乙酸乙酯溶解固体(30mL×3)，过滤，合并后用饱和NaCl溶液洗涤(60mL×3)。无水MgSO₄干燥、旋蒸除溶剂、干燥，得到淡黄色固体，收率68.9％。¹H NMR(400MHz,DMSO-d₆)δ14.51(s,1H),8.84(s,1H),8.45(s,1H).ESI-MS m/z:153[M-H]^-。In a 100mL three-necked flask, add 1.0g of 4,6-dichloro-5-pyrimidinecarbaldehyde and 20mL of methanol, stir to dissolve, cool down to -65°C, and dropwise add 0.97mL (1.2eq) of triethylamine. Dilute 0.274mL (1.0eq) of hydrazine monohydrate with 10mL of methanol, and drop it slowly with a dropping funnel. Return to room temperature after dropping, react for 2-3 hours, and monitor by TLC. After the reaction was completed, the solvent was evaporated to dryness and dried. The solids were dissolved with ethyl acetate (30 mL×3), filtered, combined and washed with saturated NaCl solution (60 mL×3). Dry over anhydrous MgSO₄ , remove the solvent by rotary evaporation, and dry to obtain a light yellow solid with a yield of 68.9%.¹ H NMR (400MHz, DMSO-d₆ ) δ 14.51(s, 1H), 8.84(s, 1H), 8.45(s, 1H). ESI-MS m/z: 153[MH]⁻ .

实施例2：Example 2:

4-氯-1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶的合成Synthesis of 4-chloro-1-(2-tetrahydropyran)-1H-pyrazolo[3,4-d]pyrimidine

在100mL三口烧瓶中，加入1.88g 4-氯-1H-吡唑并[3,4-d]嘧啶，50mL乙酸乙酯，升温至50℃。依次投入50mg的PPTs(对甲苯磺酸吡啶盐，催化剂量)、1.37mL 3,4-二氢-2H-吡喃(1.2eq)。50℃保温反应20-22h，TLC监测。反应结束，降至室温，分别用水(60mL×1)、饱和NaCl溶液(50mL×2)洗涤混合物。无水MgSO₄干燥、旋蒸除溶剂、干燥。用石油醚萃取所得固体(60mL×2)，蒸干溶剂，干燥后得淡黄色油状固体，收率76.5％。¹H NMR(400MHz,DMSO-d₆)δ8.92(s,1H),8.55(s,1H),6.02(dd,J＝10.4,2.4Hz,1H),3.97(d,J＝12.0Hz,1H),3.76–3.70(m,1H),2.49–2.42(m,1H),2.07-2.08(m,1H),1.98–1.94(m,1H),1.85–1.73(m,1H),1.64–1.58(m,2H).ESI-MS m/z:261[M+Na]⁺。In a 100 mL three-necked flask, 1.88 g of 4-chloro-1H-pyrazolo[3,4-d]pyrimidine and 50 mL of ethyl acetate were added, and the temperature was raised to 50°C. 50 mg of PPTs (pyridinium p-toluenesulfonate, catalyst amount) and 1.37 mL of 3,4-dihydro-2H-pyran (1.2 eq) were added in sequence. The reaction was incubated at 50°C for 20-22h, monitored by TLC. After the reaction was completed, it was lowered to room temperature, and the mixture was washed with water (60 mL×1) and saturated NaCl solution (50 mL×2), respectively. Dry over anhydrous MgSO₄ , remove solvent by rotary evaporation, and dry. The resulting solid (60 mL×2) was extracted with petroleum ether, and the solvent was evaporated to dryness to obtain a light yellow oily solid with a yield of 76.5%.¹ H NMR (400MHz, DMSO-d₆ )δ8.92(s, 1H), 8.55(s, 1H), 6.02(dd, J=10.4, 2.4Hz, 1H), 3.97(d, J=12.0Hz, 1H),3.76–3.70(m,1H),2.49–2.42(m,1H),2.07-2.08(m,1H),1.98–1.94(m,1H),1.85–1.73(m,1H),1.64– 1.58 (m, 2H). ESI-MS m/z: 261 [M+Na]⁺ .

实施例3：Example 3:

4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯胺的合成Synthesis of 4-[[1-(2-tetrahydropyran)-1H-pyrazolo[3,4-d]pyrimidine]-4-oxyl]aniline

在100mL三口烧瓶中，加入0.28g对氨基酚，1.51g Cs₂CO₃(1.2eq)粉末，N₂保护下滴入2mL DMF，室温搅拌1.5-2h。后将溶于适量DMF中的0.56g 4-氯-1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶(1.0eq)滴入，反应22-24h，TLC监测。反应结束，用50mL乙酸乙酯稀释后，依次用1M的NaOH溶液(60mL×1)、水(80mL×1)、5％LiCl溶液(50mL×2)洗涤，MgSO₄干燥、旋蒸除溶剂、干燥得棕褐色油状固体，收率79.5％。¹H NMR(400MHz,DMSO-d₆)δ8.57(s,1H),7.75(s,1H),6.99–6.95(m,2H),6.67–6.31(m,2H),5.96(dd,J＝10.0,2.4Hz,1H),5.19(s,2H),3.96(d,J＝12.4Hz,1H),3.73–3.67(m,1H),2.48-2.40(m,1H),2.06-2.00(m,1H),1.92–1.88(m,1H),1.79-1.71(m,1H),1.61–1.56(m,2H).ESI-MS m/z:310[M-H]^-。In a 100mL three-neck flask, add 0.28g of p-aminophenol, 1.51g of Cs₂ CO₃ (1.2eq) powder, drop into 2mL of DMF under the protection of N₂ , and stir at room temperature for 1.5-2h. Then, 0.56g of 4-chloro-1-(2-tetrahydropyran)-1H-pyrazolo[3,4-d]pyrimidine (1.0eq) dissolved in an appropriate amount of DMF was added dropwise and reacted for 22-24h. TLC monitoring. After the reaction was completed, after diluting with 50 mL of ethyl acetate, wash with 1M NaOH solution (60 mL×1), water (80 mL×1), 5% LiCl solution (50 mL×2) successively, MgSO₄ Dry, rotary evaporate to remove solvent, After drying, a tan oily solid was obtained with a yield of 79.5%.¹ H NMR (400MHz,DMSO-d₆ )δ8.57(s,1H),7.75(s,1H),6.99–6.95(m,2H),6.67–6.31(m,2H),5.96(dd,J =10.0,2.4Hz,1H),5.19(s,2H),3.96(d,J=12.4Hz,1H),3.73–3.67(m,1H),2.48-2.40(m,1H),2.06-2.00( m, 1H), 1.92–1.88(m, 1H), 1.79-1.71(m, 1H), 1.61–1.56(m, 2H). ESI-MS m/z: 310[MH]^- .

实施例4：Example 4:

1-(3-甲基苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲的合成1-(3-methylphenyl)-3-[4-[[1-(2-tetrahydropyran)-1H-pyrazolo[3,4-d]pyrimidine]-4-oxyl]benzene Synthesis of urea

在100mL三口烧瓶中，加入4-氯-1H-吡唑并[3,4-d]嘧啶，适量CH₂Cl₂，冰浴冷却，滴加1.0eq量的3-甲基苯基异氰酸酯的CH₂Cl₂。撤去冰浴，室温下反应过夜，TLC监测。反应结束，加入60mL石油醚洗涤，过滤、干燥后得到粗产物，酸性氧化铝柱层析分离(乙酸乙酯/甲醇)得到目标化合物，收率80.0％。¹H NMR(400MHz,DMSO-d₆)δ14.13(s,1H),8.82(s,1H),8.66(s,1H),8.51(s,1H),8.01(s,1H),7.56(d,J＝8.9Hz,2H),7.32(s,1H),7.25(d,J＝8.9Hz,3H),7.17(t,J＝7.7Hz,1H),6.80(d,J＝7.7Hz,1H),2.29(s,3H).¹³C NMR(101MHz,DMSO-d₆)δ163.68,157.13,155.39,153.00,146.77,139.99,138.36,138.11,132.26,129.04,123.02,122.66,120.47,119.63,117.67,117.24,102.01.ESI-MS m/z:445[M+H]⁺。In a 100mL three-necked flask, add 4-chloro-1H-pyrazolo[3,4-d]pyrimidine, an appropriate amount of CH₂ Cl₂ , cool in an ice bath, add 1.0eq of 3-methylphenylisocyanate CH₂ Cl₂ . Remove the ice bath, react overnight at room temperature, and monitor by TLC. After the reaction was completed, 60 mL of petroleum ether was added to wash, filtered, and dried to obtain a crude product, which was separated by acidic alumina column chromatography (ethyl acetate/methanol) to obtain the target compound with a yield of 80.0%.¹ H NMR (400MHz,DMSO-d₆ )δ14.13(s,1H),8.82(s,1H),8.66(s,1H),8.51(s,1H),8.01(s,1H),7.56( d,J=8.9Hz,2H),7.32(s,1H),7.25(d,J=8.9Hz,3H),7.17(t,J=7.7Hz,1H),6.80(d,J=7.7Hz, 1H),2.29(s,3H).¹³ C NMR(101MHz,DMSO-d₆ )δ163.68,157.13,155.39,153.00,146.77,139.99,138.36,138.11,132.26,129.04,123.02,122.66,120.46 , 117.24, 102.01. ESI-MS m/z: 445 [M+H]⁺ .

实施5：Implementation 5:

1-(3,4-二氯苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲1-(3,4-Dichlorophenyl)-3-[4-[[1-(2-tetrahydropyran)-1H-pyrazolo[3,4-d]pyrimidine]-4-oxyl ]phenyl]urea

参照实施例4的制备方法。收率67.0％。¹H NMR(400MHz,DMSO-d₆)δ9.07(s,1H),8.97(s,1H),8.58(s,1H),8.14(s,1H),7.90(s,1H),7.58–7.52(m,3H),7.37(d,J＝8.4Hz,1H),7.27 (d,J＝8.8Hz,2H),5.99(d,J＝9.6Hz,1H),3.97(d,J＝10.8Hz,1H),3.71(s,1H),2.05(d,J＝12.8Hz,1H),1.93(d,J＝12.4Hz,1H),1.79(s,1H),1.59(s,2H),1.24(s,1H).¹³CNMR(101MHz,DMSO-d₆)δ163.71,155.73,152.81,147.00,140.36,137.71,132.26,131.45,131.00,123.56,122.66,120.18,119.76,118.82,102.73,82.70,67.55,29.14,25.02,22.59.ESI-MS m/z:499[M+H]⁺。Referring to the preparation method of Example 4. Yield 67.0%.¹ H NMR (400MHz,DMSO-d₆ )δ9.07(s,1H),8.97(s,1H),8.58(s,1H),8.14(s,1H),7.90(s,1H),7.58– 7.52(m,3H),7.37(d,J=8.4Hz,1H),7.27(d,J=8.8Hz,2H),5.99(d,J=9.6Hz,1H),3.97(d,J=10.8 Hz,1H),3.71(s,1H),2.05(d,J=12.8Hz,1H),1.93(d,J=12.4Hz,1H),1.79(s,1H),1.59(s,2H), 1.24 (s, 1H).¹³ CNMR (101MHz, DMSO-D₆ ) Δ163.71,155.73,152.81,147.00,140.36,1326,131.45,123.56,18.76,76,72,72,72,72,72,2.72.72.72.72.72.72.72.72.72.72. , 29.14, 25.02, 22.59. ESI-MS m/z: 499 [M+H]⁺ .

实施例6：Embodiment 6:

1-(2-氯-5-甲基苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲1-(2-Chloro-5-methylphenyl)-3-[4-[[1-(2-tetrahydropyran)-1H-pyrazolo[3,4-d]pyrimidine]-4- Oxy]phenyl]urea

参照实施例4的制备方法。收率69.4％。¹H NMR(400MHz,DMSO-d₆)δ9.54(s,1H),8.59(s,1H),8.28(s,1H),8.13(s,1H),8.03(s,1H),7.58(d,J＝8.8Hz,2H),7.34(d,J＝8.0Hz,1H),7.28(d,J＝8.8Hz,2H),6.87(d,J＝8.0Hz,1H),5.99(d,J＝10.0Hz,1H),3.97(d,J＝11.2Hz,1H),3.75–3.68(m,1H),2.30(s,3H),2.08–2.00(m,1H),1.93(d,J＝11.6Hz,1H),1.77(s,1H),1.60(s,2H),1.24(s,1H).¹³C NMR(101MHz,DMSO-d₆)δ163.71,155.73,155.68,152.58,146.87,137.90,137.47,135.92,132.26,129.20,124.47,122.72,122.20,119.75,119.45,102.72,82.70,67.54,29.14,25.02,22.59,21.32.ESI-MS m/z:477[M-H]^-。Referring to the preparation method of Example 4. Yield 69.4%.¹ H NMR (400MHz,DMSO-d₆ )δ9.54(s,1H),8.59(s,1H),8.28(s,1H),8.13(s,1H),8.03(s,1H),7.58( d,J=8.8Hz,2H),7.34(d,J=8.0Hz,1H),7.28(d,J=8.8Hz,2H),6.87(d,J=8.0Hz,1H),5.99(d, J＝10.0Hz, 1H), 3.97(d, J＝11.2Hz, 1H), 3.75–3.68(m, 1H), 2.30(s, 3H), 2.08–2.00(m, 1H), 1.93(d,J ＝11.6Hz, 1H), 1.77(s, 1H), 1.60(s, 2H), 1.24(s, 1H).¹³ C NMR (101MHz, DMSO-d₆ ) δ163.71, 155.73, 155.68, 152.58, 146.87, 137.90 ,^{137.47,135.92,132.26,129.20,124.47,122.72,122.20,119.75,119.45,102.72,82.70,67.54,29.14,25.02,22.59,21.32} .

实施例7：Embodiment 7:

1-(2,3-二甲基苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲1-(2,3-Dimethylphenyl)-3-[4-[[1-(2-tetrahydropyran)-1H-pyrazolo[3,4-d]pyrimidine]-4-oxo base]phenyl]urea

参照实施例4的制备方法。收率65.5％。¹H NMR(400MHz,DMSO-d₆)δ9.07(s,1H),8.58(s,1H),8.10(s,1H),8.00(s,1H),7.56(t,J＝8.6Hz,3H),7.25(d,J＝8.4Hz,1H),7.05(t,J＝7.8Hz,1H),6.92(d,J＝7.2Hz,1H),5.99(d,J＝9.2Hz,1H),3.97(d,J＝11.2Hz,1H),3.75-3.68(m,1H),2.45(d,J＝10.4Hz,1H),2.27(s,3H),2.16(s,3H),2.05(d,J＝10.8Hz,1H),1.93(d,J＝12.0Hz,1H),1.79(s,1H),1.60(s,2H),1.46(s,1H).¹³C NMR(101MHz,DMSO-d₆))δ155.74,153.38, 146.51,138.49,137.31,137.00,132.26,128.07,125.65,125.42,122.60,120.92,119.50,102.70,82.69,67.54,29.14,25.02,22.59,20.75,14.05.ESI-MSm/z:481[M+Na]⁺。Referring to the preparation method of Example 4. Yield 65.5%.¹ H NMR (400MHz, DMSO-d₆ )δ9.07(s,1H),8.58(s,1H),8.10(s,1H),8.00(s,1H),7.56(t,J＝8.6Hz, 3H), 7.25(d, J=8.4Hz, 1H), 7.05(t, J=7.8Hz, 1H), 6.92(d, J=7.2Hz, 1H), 5.99(d, J=9.2Hz, 1H) ,3.97(d,J=11.2Hz,1H),3.75-3.68(m,1H),2.45(d,J=10.4Hz,1H),2.27(s,3H),2.16(s,3H),2.05( d, J=10.8Hz, 1H), 1.93(d, J=12.0Hz, 1H), 1.79(s, 1H), 1.60(s, 2H), 1.46(s, 1H).¹³ C NMR (101MHz, DMSO -d₆ ))δ155.74,153.38, 146.51,138.49,137.31,137.00,132.26,128.07,125.65,125.42,122.60,120.92,119.50,102.70,82.69,67.54,29.14,25.02,22.59,20.75,14.05.ESI-MSm /z:481[M+Na]⁺ .

实施例8：Embodiment 8:

1-(4-氟-3-甲基苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲1-(4-fluoro-3-methylphenyl)-3-[4-[[1-(2-tetrahydropyran)-1H-pyrazolo[3,4-d]pyrimidine]-4- Oxy]phenyl]urea

参照实施例4的制备方法。收率79.4％。¹H NMR(400MHz,DMSO-d₆)δ8.83(s,1H),8.69(s,1H),8.58(s,1H),8.11(s,1H),7.56(d,J＝8.8Hz,2H),7.38(dd,J＝6.8,2.4Hz,1H),7.30-7.24(m,3H),7.06(t,J＝9.2Hz,1H),5.98(dd,J＝10.4,2.4Hz,1H),3.97(d,J＝10.8Hz,1H),3.74–3.658(m,1H),2.22(s,3H),2.05(d,J＝11.6Hz,1H),1.95–1.90(m,1H),1.80-1.76(m,1H),1.59(s,2H),1.24(s,1H).¹³C NMR(101MHz,DMSO-d₆)δ163.76,155.70,155.54,153.17,146.71,137.99,135.73,132.25,124.79,124.61,122.58,121.94,120.04,118.10,118.03,115.44,115.21,102.65,82.68,67.65,29.10,24.92,22.54,14.72.ESI-MSm/z:485[M+Na]⁺。Referring to the preparation method of Example 4. Yield 79.4%.¹ H NMR (400MHz, DMSO-d₆ )δ8.83(s,1H),8.69(s,1H),8.58(s,1H),8.11(s,1H),7.56(d,J＝8.8Hz, 2H), 7.38(dd, J=6.8, 2.4Hz, 1H), 7.30-7.24(m, 3H), 7.06(t, J=9.2Hz, 1H), 5.98(dd, J=10.4, 2.4Hz, 1H ),3.97(d,J=10.8Hz,1H),3.74–3.658(m,1H),2.22(s,3H),2.05(d,J=11.6Hz,1H),1.95–1.90(m,1H) ,1.80-1.76(m,1H),1.59(s,2H),1.24(s,1H).¹³ C NMR(101MHz,DMSO-d₆ )δ163.76,155.70,155.54,153.17,146.71,137.99,135.73,132.25 ,^{124.79,124.61,122.58,121.94,120.04,118.10,118.03,115.44,115.21,102.65,82.68,67.65,29.10,24.92,22.54,14.72} .

实施例9：Embodiment 9:

1-(3-硝基苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲1-(3-nitrophenyl)-3-[4-[[1-(2-tetrahydropyran)-1H-pyrazolo[3,4-d]pyrimidine]-4-oxyl]benzene base] urea

参照实施例4的制备方法。收率62.3％。¹H NMR(400MHz,DMSO-d₆)δ9.41(s,1H),9.11(s,1H),8.59(s,2H),8.16(s,1H),7.85(s,1H),7.74(s,1H),7.59(s,2H),7.29–7.00(m,2H),5.99(d,J＝12.4Hz,1H),5.31(s,1H),3.96(s,2H),3.72(s,2H),2.02(s,2H),1.77(s,1H),1.59(s,1H).¹³C NMR(101MHz,DMSO-d₆)δ167.36,163.69,155.70,152.91,148.58,147.07,141.45,137.66,132.23,131.95,130.46,129.04,124.77,122.64,120.27,116.72,112.61,102.74,82.72,67.82,28.77,22.79.ESI-MS m/z:474[M-H]^-。Referring to the preparation method of Example 4. Yield 62.3%.¹ H NMR (400MHz,DMSO-d₆ )δ9.41(s,1H),9.11(s,1H),8.59(s,2H),8.16(s,1H),7.85(s,1H),7.74( s,1H),7.59(s,2H),7.29–7.00(m,2H),5.99(d,J=12.4Hz,1H),5.31(s,1H),3.96(s,2H),3.72(s ,2H),2.02(s,2H),1.77(s,1H),1.59(s,1H).¹³ C NMR(101MHz,DMSO-d₆ )δ167.36,163.69,155.70,152.91,148.58,147.07,141.45, 137.66, 132.23, 131.95, 130.46, 129.04, 124.77, 122.64, 120.27, 116.72, 112.61, 102.74, 82.72, 67.82, 28.77, 22.79. ESI-MS m/z: 474[MH]^- .

实施例10：Example 10:

1-(2-氟-5-(三氟甲基)苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲1-(2-fluoro-5-(trifluoromethyl)phenyl)-3-[4-[[1-(2-tetrahydropyran)-1H-pyrazolo[3,4-d]pyrimidine ]-4-oxyl]phenyl]urea

参照实施例4的制备方法。收率70.2％。¹H NMR(400MHz,DMSO-d₆)δ9.35(s,1H),8.96(s,1H),8.61(d,J＝22.8Hz,2H),8.16(s,1H),7.55(d,J＝26.4Hz,3H),7.41(s,1H),7.29(d,J＝5.6Hz,2H),5.98(d,J＝8.8Hz,1H),3.95(s,1H),3.71(s,1H),2.03(s,1H),1.92(d,J＝12.4Hz,2H),1.78(s,1H),1.59(s,2H).¹³C NMR(101MHz,DMSO-d₆)δ163.65,155.73,155.64,152.54,147.08,137.40,132.27,129.13,128.99,122.79,119.92,116.94,116.62,116.39,115.63,102.71,82.65,67.55,29.13,25.01,22.60.ESI-MS m/z:517[M+H]⁺。Referring to the preparation method of Example 4. Yield 70.2%.¹ H NMR (400MHz, DMSO-d₆ )δ9.35(s, 1H), 8.96(s, 1H), 8.61(d, J=22.8Hz, 2H), 8.16(s, 1H), 7.55(d, J＝26.4Hz, 3H), 7.41(s, 1H), 7.29(d, J＝5.6Hz, 2H), 5.98(d, J＝8.8Hz, 1H), 3.95(s, 1H), 3.71(s, 1H), 2.03(s, 1H), 1.92(d, J=12.4Hz, 2H), 1.78(s, 1H), 1.59(s, 2H).¹³ C NMR(101MHz, DMSO-d₆ ) δ163.65, 155.73 ,155.64,152.54,147.08,137.40,132.27,129.13,128.99,122.79,119.92,116.94,116.62,116.39,115.63,102.71,82.65,67.55,29.13,25.01/ES m H]⁺ .

实施例11：Example 11:

1-(3-氰基苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲1-(3-cyanophenyl)-3-[4-[[1-(2-tetrahydropyran)-1H-pyrazolo[3,4-d]pyrimidine]-4-oxyl]benzene base] urea

参照实施例4的制备方法。收率69.8％。¹H NMR(400MHz,DMSO-d₆)δ9.10(s,1H),9.00(s,1H),8.58(s,1H),8.13(s,1H),8.00(t,J＝1.8Hz,1H),7.71(ddd,J＝3.2,2.0,1.2Hz,1H),7.59(d,J＝2.4Hz,1H),7.57(d,J＝2.0Hz,1H),7.51(t,J＝8.0Hz,1H),7.43(dt,J＝7.6,1.2Hz,1H),7.29–7.27(m,2H),5.99(dd,J＝10.0,2.4Hz,1H),3.18(d,J＝5.2Hz,2H),1.99(s,6H).¹³C NMR(101MHz,DMSO-d₆)δ163.68,155.73,152.89,146.93,140.99,137.71,132.27,130.61,125.76,123.33,122.68,121.18,120.12,119.29,111.98,102.70,82.65,67.56,60.18,29.13,25.01,22.60,21.17,14.48.ESI-MS m/z:456[M+H]⁺。Referring to the preparation method of Example 4. Yield 69.8%.¹ H NMR (400MHz,DMSO-d₆ )δ9.10(s,1H),9.00(s,1H),8.58(s,1H),8.13(s,1H),8.00(t,J＝1.8Hz, 1H), 7.71(ddd, J=3.2, 2.0, 1.2Hz, 1H), 7.59(d, J=2.4Hz, 1H), 7.57(d, J=2.0Hz, 1H), 7.51(t, J=8.0 Hz, 1H), 7.43(dt, J=7.6, 1.2Hz, 1H), 7.29–7.27(m, 2H), 5.99(dd, J=10.0, 2.4Hz, 1H), 3.18(d, J=5.2Hz ,2H),1.99(s,6H).¹³ C NMR(101MHz,DMSO-d₆ )δ163.68,155.73,152.89,146.93,140.99,137.71,132.27,130.61,125.76,123.33,122.68,121.12,110. 111.98, 102.70, 82.65, 67.56, 60.18, 29.13, 25.01, 22.60, 21.17, 14.48. ESI-MS m/z: 456[M+H]⁺ .

实施例12：Example 12:

1-(5-甲基-2-硝基苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲1-(5-Methyl-2-nitrophenyl)-3-[4-[[1-(2-tetrahydropyran)-1H-pyrazolo[3,4-d]pyrimidine]-4 -oxy]phenyl]urea

参照实施例4的制备方法。收率61.8％。¹H NMR(400MHz,DMSO-d₆)δ10.05(s,1H),9.71(s,1H),8.58(s,1H),8.22(s,1H),8.14(s,1H),8.04(d,J＝8.4Hz,1H),7.61(d,J＝8.8Hz,2H),7.29(d,J＝8.4Hz,2H),7.03(d,J＝8.8Hz,1H),5.98(d,J＝9.6Hz,1H),3.97(d,J＝11.2Hz,1H),3.71(dd,J＝12.8,6.4Hz,1H),2.40(s,4H),2.03(s,1H),1.92(d,J＝12.0Hz,1H),1.78(s,1H),1.59(s,2H).¹³C NMR(101MHz,DMSO-d₆)δ163.70,155.76,155.70,152.24,147.09,146.76,137.68,135.52,132.28,125.95,123.53,122.76,122.58,120.21,102.74,82.70,67.60,29.14,25.02,22.60,21.94.ESI-MS m/z:488[M-H]^-。Referring to the preparation method of Example 4. Yield 61.8%.¹ H NMR (400MHz,DMSO-d₆ )δ10.05(s,1H),9.71(s,1H),8.58(s,1H),8.22(s,1H),8.14(s,1H),8.04( d,J=8.4Hz,1H),7.61(d,J=8.8Hz,2H),7.29(d,J=8.4Hz,2H),7.03(d,J=8.8Hz,1H),5.98(d, J＝9.6Hz, 1H), 3.97(d, J＝11.2Hz, 1H), 3.71(dd, J＝12.8, 6.4Hz, 1H), 2.40(s, 4H), 2.03(s, 1H), 1.92( d, J＝12.0Hz, 1H), 1.78(s, 1H), 1.59(s, 2H).¹³ C NMR (101MHz, DMSO-d₆ ) δ163.70, 155.76, 155.70, 152.24, 147.09, 146.76, 137.68, 135.52 ,132.28,125.95,123.53,122.76,122.58,120.21,102.74,82.70,67.60,29.14,25.02,22.60,21.94. ESI-MS m/z: 488[MH]^- .

实施例13：Example 13:

1-(2-甲基苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲1-(2-methylphenyl)-3-[4-[[1-(2-tetrahydropyran)-1H-pyrazolo[3,4-d]pyrimidine]-4-oxyl]benzene base] urea

参照实施例4的制备方法。收率74.5％。¹H NMR(400MHz,DMSO-d₆)δ8.41(s,1H),7.95(s,1H),7.81(t,J＝8.4Hz,2H),7.49(d,J＝7.2Hz,1H),7.36(t,J＝7.6Hz,1H),7.19(t,J＝7.6Hz,1H),7.00(s,3H),6.89(t,J＝7.8Hz,2H),6.74(d,J＝8.0Hz,1H),5.37(d,J＝6.0Hz,2H),4.54(s,3H),3.05(s,1H),1.67(d,J＝5.2Hz,5H).¹³C NMR(101MHz,DMSO-d₆)δ167.01,159.65,151.08,141.82,141.36,140.59,135.03,133.47,129.50,128.93,128.22,126.97,125.17,123.42,122.12,119.87,113.33,112.36,55.14,43.52,24.14.ESI-MS m/z:443[M-H]^-。Referring to the preparation method of Example 4. Yield 74.5%.¹ H NMR (400MHz,DMSO-d₆ )δ8.41(s,1H),7.95(s,1H),7.81(t,J=8.4Hz,2H),7.49(d,J=7.2Hz,1H) ,7.36(t,J=7.6Hz,1H),7.19(t,J=7.6Hz,1H),7.00(s,3H),6.89(t,J=7.8Hz,2H),6.74(d,J= 8.0Hz, 1H), 5.37(d, J＝6.0Hz, 2H), 4.54(s, 3H), 3.05(s, 1H), 1.67(d, J＝5.2Hz, 5H).¹³ C NMR (101MHz, DMSO-d₆ )δ167.01,159.65,151.08,141.82,141.36,140.59,135.03,133.47,129.50,128.93,128.22,126.97,125.17,123.42,122.12,119.87,113.33,112.36,55.14,43.52,24.14.ESI-MS m/z:443[MH]^- .

实施例14：Example 14:

1-(3-氟苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲1-(3-fluorophenyl)-3-[4-[[1-(2-tetrahydropyran)-1H-pyrazolo[3,4-d]pyrimidine]-4-oxyl]phenyl ] urea

参照实施例4的制备方法。收率61.2％。¹H NMR(400MHz,DMSO-d₆)δ8.96(s,1H),8.89(s,1H),8.58(s,1H),8.13(s,1H),7.57(d,J＝8.8Hz,2H),7.51(dt,J＝11.6,2.4Hz,1H),7.35–7.26(m,3H),7.15(d,J＝8.0Hz,1H),6.80(td,J＝8.4,2.4Hz,1H),5.98(dd,J＝10.0,2.4Hz,1H),3.97(d,J＝11.2Hz,1H),3.71(ddd,J＝11.6,6.4,4.8Hz,1H),2.06–2.03(m,1H),1.95–1.91(m,1H),1.80–1.72(m,1H),1.63–1.57(m,2H),1.23(s,1H).¹³C NMR(101MHz,DMSO-d₆)δ163.71,155.72,152.85,146.87,142.00,137.84,132.24,130.77,130.67,122.63,120.02,116.25,114.39,114.36,108.69,108.48,105.44,102.72,82.69,67.54,29.14,25.02,22.59,21.52.ESI-MS m/z:447[M-H]^-。Referring to the preparation method of Example 4. Yield 61.2%.¹ H NMR (400MHz, DMSO-d₆ )δ8.96(s,1H),8.89(s,1H),8.58(s,1H),8.13(s,1H),7.57(d,J＝8.8Hz, 2H), 7.51(dt, J=11.6, 2.4Hz, 1H), 7.35–7.26(m, 3H), 7.15(d, J=8.0Hz, 1H), 6.80(td, J=8.4, 2.4Hz, 1H ),5.98(dd,J=10.0,2.4Hz,1H),3.97(d,J=11.2Hz,1H),3.71(ddd,J=11.6,6.4,4.8Hz,1H),2.06–2.03(m, 1H),1.95–1.91(m,1H),1.80–1.72(m,1H),1.63–1.57(m,2H),1.23(s,1H).¹³ C NMR(101MHz,DMSO-d₆ )δ163. 71,155.72,152.85,146.87,142.00,137.84,132.24,130.77,130.67,122.63,120.02,116.25,114.39,114.36,108.69,108.48,105.44,102.72,82.69,67.54,29.14,25.02,22.59,21.52.ESI-MS m /z:447[MH]^- .

实施例15：Example 15:

1-(4-氟苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲1-(4-fluorophenyl)-3-[4-[[1-(2-tetrahydropyran)-1H-pyrazolo[3,4-d]pyrimidine]-4-oxyl]phenyl ] urea

参照实施例4的制备方法。收率66.3％。¹H NMR(400MHz,CDCl₃)δ8.57(s,1H),7.88(s,1H),7.59(d,J＝7.6Hz,4H),7.54(d,J＝7.7Hz,4H),7.49(s,2H),7.41(d,J＝8.5Hz,4H),7.20(d,J＝7.1Hz,13H),7.13–7.07(m,13H),6.56(s,3H),6.06(d,J＝9.1Hz,1H),4.14(d,J＝11.3Hz,2H),3.89–3.77(m,2H),2.22(d,J＝9.3Hz,13H).¹³C NMR(101MHz,DMSO-d₆)δ167.35,163.77,155.72,153.11,146.61,138.36,137.74,132.14,131.93,130.58,129.04,128.06,126.55,122.61,121.57,119.56,102.72,82.71,67.81,28.77,22.79,14.26.ESI-MS m/z:447[M-H]^-。Referring to the preparation method of Example 4. Yield 66.3%.¹ H NMR (400MHz, CDCl₃ )δ8.57(s,1H),7.88(s,1H),7.59(d,J=7.6Hz,4H),7.54(d,J=7.7Hz,4H),7.49 (s,2H),7.41(d,J=8.5Hz,4H),7.20(d,J=7.1Hz,13H),7.13–7.07(m,13H),6.56(s,3H),6.06(d, J＝9.1Hz, 1H), 4.14(d, J＝11.3Hz, 2H), 3.89–3.77(m, 2H), 2.22(d, J＝9.3Hz, 13H).¹³ C NMR (101MHz, DMSO-d₆ )δ167.35,163.77,155.72,153.11,146.61,138.36,137.74,132.14,131.93,130.58,129.04,128.06,126.55,122.61,121.57,119.56,102.72,82.71,67.81,28.77,22.79,14.26.ESI-MS m /z:447[MH]^- .

实施例16：Example 16:

1-(2,5-二甲基苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲1-(2,5-Dimethylphenyl)-3-[4-[[1-(2-tetrahydropyran)-1H-pyrazolo[3,4-d]pyrimidine]-4-oxo base]phenyl]urea

参照实施例4的制备方法。收率71.2％。¹H NMR(400MHz,DMSO-d₆)δ9.19(s,1H),8.59(s,1H),8.12(s,1H),7.93(s,1H),7.70(s,1H),7.58(d,J＝6.8Hz,2H),7.26(d,J＝6.7Hz,2H),7.05(s,1H),6.77(s,1H),5.98(d,J＝9.2Hz,1H),3.97(d,J＝10.0Hz,1H),3.71(s,1H),2.24(d,J＝9.2Hz,6H),2.02(s,1H),1.92(d,J＝12.8Hz,1H),1.78(s,1H),1.59(s,3H).¹³C NMR(101MHz,DMSO-d₆)δ163.76,155.75,155.64,153.09,146.52,138.36,137.50,135.49,132.27,130.40,124.87,123.80,122.65,122.06,119.49,102.69,99.92,82.65,67.56,29.14,25.01,22.60,21.35,17.90.ESI-MS m/z:457[M-H]^-。Referring to the preparation method of Example 4. Yield 71.2%.¹ H NMR (400MHz,DMSO-d₆ )δ9.19(s,1H),8.59(s,1H),8.12(s,1H),7.93(s,1H),7.70(s,1H),7.58( d,J=6.8Hz,2H),7.26(d,J=6.7Hz,2H),7.05(s,1H),6.77(s,1H),5.98(d,J=9.2Hz,1H),3.97( d,J=10.0Hz,1H),3.71(s,1H),2.24(d,J=9.2Hz,6H),2.02(s,1H),1.92(d,J=12.8Hz,1H),1.78( s,1H),1.59(s,3H).¹³ C NMR(101MHz,DMSO-d₆ )δ163.76,155.75,155.64,153.09,146.52,138.36,137.50,135.49,132.27,130.40,124.857,123.860,112.2 , 119.49, 102.69, 99.92, 82.65, 67.56, 29.14, 25.01, 22.60, 21.35, 17.90. ESI-MS m/z: 457[MH]^- .

实施例17：Example 17:

1-(3-三氟甲基苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲1-(3-Trifluoromethylphenyl)-3-[4-[[1-(2-tetrahydropyran)-1H-pyrazolo[3,4-d]pyrimidine]-4-oxyl ]phenyl]urea

参照实施例4的制备方法。收率73.2％。¹H NMR(400MHz,DMSO-d₆)δ9.17(s,1H),9.01(s,1H),8.58(s,1H),8.13(s,1H),8.03(s,1H),7.62–7.51(m,4H),7.30(dd,J＝20.0,7.6Hz,3H),5.98(d,J＝9.2Hz,1H),3.97(d,J＝11.2Hz,1H),3.70(s,1H),2.05–1.99(m,1H),1.92(d,J＝11.6Hz,1H),1.76(s,1H),1.59(s,3H).¹³C NMR(101MHz,DMSO-d₆)δ163.70,155.72,155.64,152.99,146.90,140.95,137.78,132.26,130.32,122.64,122.23,121.17,120.11,118.53,115.57,114.53,102.70,82.65,67.56,29.13,25.01,22.59.ESI-MS m/z:497[M-H]^-。Referring to the preparation method of Example 4. Yield 73.2%.¹ H NMR (400MHz,DMSO-d₆ )δ9.17(s,1H),9.01(s,1H),8.58(s,1H),8.13(s,1H),8.03(s,1H),7.62– 7.51(m,4H),7.30(dd,J=20.0,7.6Hz,3H),5.98(d,J=9.2Hz,1H),3.97(d,J=11.2Hz,1H),3.70(s,1H ),2.05–1.99(m,1H),1.92(d,J＝11.6Hz,1H),1.76(s,1H),1.59(s,3H).¹³ C NMR(101MHz,DMSO-d₆ )δ163. 70,155.72,155.64,152.99,146.90,140.95,137.78,132.26,130.32,122.64,122.23,121.17,120.11,118.53,115.57,114.53,102.70,82.65,67.56,29.13,25.01,22.59.ESI-MS m/z:497 [MH]^- .

实施例18：Example 18:

1-(3-三氟甲基-4-氯苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲1-(3-Trifluoromethyl-4-chlorophenyl)-3-[4-[[1-(2-tetrahydropyran)-1H-pyrazolo[3,4-d]pyrimidine]- 4-Oxy]phenyl]urea

参照实施例4的制备方法。收率66.6％。¹H NMR(400MHz,DMSO-d₆)δ9.22(s,1H),8.99(s,1H),8.58(s,1H),8.14(s,2H),7.69–7.57(m,4H),7.27(d,J＝8.8Hz,2H),5.98(d,J＝10.0Hz,1H),3.97(d,J＝11.6Hz,1H),3.74-3.68(m,1H),2.05(d,J＝12.4Hz,1H),1.93(d,J＝12.4Hz,1H),1.77(d,J＝8.0Hz,1H),1.59(s,3H).¹³C NMR(101MHz,DMSO-d₆)δ163.7,155.7,152.9,147.1,139.8,137.6,132.4,132.3,127.3,123.5,122.7,121.9,120.3,117.3,102.7,82.7,67.5,29.1,25.0,22.6.ESI-MS m/z:531[M-H]^-。Referring to the preparation method of Example 4. Yield 66.6%.¹ H NMR (400MHz,DMSO-d₆ )δ9.22(s,1H),8.99(s,1H),8.58(s,1H),8.14(s,2H),7.69–7.57(m,4H), 7.27(d,J=8.8Hz,2H),5.98(d,J=10.0Hz,1H),3.97(d,J=11.6Hz,1H),3.74-3.68(m,1H),2.05(d,J ＝12.4Hz, 1H), 1.93(d, J＝12.4Hz, 1H), 1.77(d, J＝8.0Hz, 1H), 1.59(s, 3H).¹³ C NMR (101MHz, DMSO-d₆ ) δ163 .7,155.7,152.9,147.1,139.8,137.6,132.4,132.3,127.3,123.5,122.7,121.9,120.3,117.3,102.7,82.7,67.5,29.1,25.0,22.6.ESI-MS m/z:531[MH^- .

实施例19：Example 19:

1-(3-氟-5-三氟甲基苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲1-(3-fluoro-5-trifluoromethylphenyl)-3-[4-[[1-(2-tetrahydropyran)-1H-pyrazolo[3,4-d]pyrimidine]- 4-Oxy]phenyl]urea

参照实施例4的制备方法。收率60.2％。¹H NMR(400MHz,DMSO-d₆)δ9.31(s,1H),9.06(s,1H),8.58(s,1H),8.15(s,1H),7.73(s,1H),7.64(d,J＝11.2Hz,1H)7.58(d,J＝8.8Hz,2H),7.28(d,J＝8.8Hz,2H),7.24(d,J＝8.5Hz,1H),5.99(dd,J＝10.1,2.1Hz,1H),3.97(d,J＝11.6Hz,1H),3.76-3.66(m,1H),2.45(dd,J＝12.8,4.0Hz,1H),2.05(d,J＝12.5Hz,1H),1.93(dd,J＝13.0,2.6Hz,1H),1.84-1.70(m,1H),1.61(d,J＝10.0Hz,2H).ESI-MS m/z:517[M+H]⁺。Referring to the preparation method of Example 4. Yield 60.2%.¹ H NMR (400MHz,DMSO-d₆ )δ9.31(s,1H),9.06(s,1H),8.58(s,1H),8.15(s,1H),7.73(s,1H),7.64( d,J=11.2Hz,1H)7.58(d,J=8.8Hz,2H),7.28(d,J=8.8Hz,2H),7.24(d,J=8.5Hz,1H),5.99(dd,J =10.1,2.1Hz,1H),3.97(d,J=11.6Hz,1H),3.76-3.66(m,1H),2.45(dd,J=12.8,4.0Hz,1H),2.05(d,J= 12.5Hz, 1H), 1.93(dd, J＝13.0, 2.6Hz, 1H), 1.84-1.70(m, 1H), 1.61(d, J＝10.0Hz, 2H).ESI-MS m/z: 517[ M+H]⁺ .

实施例20：Example 20:

1-(4-氯苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲1-(4-Chlorophenyl)-3-[4-[[1-(2-tetrahydropyran)-1H-pyrazolo[3,4-d]pyrimidine]-4-oxyl]phenyl ] urea

参照实施例4的制备方法。收率68.3％。1H NMR(400MHz,DMSO-d6)δ8.87(s,1H),8.85(s,1H),8.58(s,1H),8.13(s,1H),7.56(d,J＝8.9Hz,2H),7.51(d,J＝8.8Hz,2H),7.26(d,J＝8.8Hz,2H),7.26(d,J＝8.9Hz,2H),5.98(dd,J＝10.1,1.9Hz,1H),3.97(d,J＝11.2Hz,1H),3.77-3.65(m,1H),2.49-2.41(m,1H),2.05(d,J＝12.5Hz,1H),1.93(dd,J＝12.9,2.3Hz,1H),1.83-1.68(m,1H),1.67-1.53(m,2H).13C NMR(101MHz,DMSO-d6)163.72,155.72,155.68,152.90,146.81,139.08,137.97,132.25,129.03,125.80,122.62,120.18,119.94,102.72,82.70,67.54,29.14,25.03,22.59.13C NMR(101MHz,DMSO)δ164.56,163.86,163.67,161.44,155.68,152.80,147.51,147.15,143.00,137.51,132.23,122.64,120.42,114.97,114.77,110.95,109.05,108.79,102.73,82.70,67.53,29.14,25.02,22.59.ESI-MS m/z:465[M+H]⁺。Referring to the preparation method of Example 4. Yield 68.3%. 1H NMR(400MHz,DMSO-d6)δ8.87(s,1H),8.85(s,1H),8.58(s,1H),8.13(s,1H),7.56(d,J＝8.9Hz,2H) ,7.51(d,J=8.8Hz,2H),7.26(d,J=8.8Hz,2H),7.26(d,J=8.9Hz,2H),5.98(dd,J=10.1,1.9Hz,1H) ,3.97(d,J=11.2Hz,1H),3.77-3.65(m,1H),2.49-2.41(m,1H),2.05(d,J=12.5Hz,1H),1.93(dd,J=12.9 ,2.3Hz,1H),1.83-1.68(m,1H),1.67-1.53(m,2H).13C NMR(101MHz,DMSO-d6)163.72,155.72,155.68,152.90,146.81,139.08,137.97,132.25, 129.03,125.80,122.62,120.18,119.94,102.72,82.70,67.54,29.14,25.03,22.59.13C NMR(101MHz,DMSO)δ164.56,163.86,163.67,161.44,155.68,152.80,147.51,147.15,143.00,137.51,132.23 ,122.64,120.42,114.97,114.77,110.95,109.05,108.79,102.73,82.70,67.53,29.14,25.02,22.59. ESI-MS m/z: 465[M+H]⁺ .

实施例21：Example 21:

1-(2-氯-5-三氟甲基苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲1-(2-Chloro-5-trifluoromethylphenyl)-3-[4-[[1-(2-tetrahydropyran)-1H-pyrazolo[3,4-d]pyrimidine]- 4-Oxy]phenyl]urea

参照实施例4的制备方法。收率74.5％。1H NMR(400MHz,DMSO-d6)δ9.22(s,1H),8.99(s,1H),8.58(s,1H),8.14(s,2H),7.69–7.57(m,4H),7.27(d,J＝8.8Hz,2H),5.98(d,J＝10.0Hz,1H),3.97(d,J＝11.6Hz,1H),3.74-3.68(m,1H),2.05(d,J＝12.4Hz,1H),1.93(d,J＝12.4Hz,1H),1.77(d,J＝8.0Hz,1H),1.59(s,3H).ESI-MS m/z:533[M+H]⁺。Referring to the preparation method of Example 4. Yield 74.5%. 1H NMR (400MHz, DMSO-d6) δ9.22(s,1H),8.99(s,1H),8.58(s,1H),8.14(s,2H),7.69–7.57(m,4H),7.27( d,J=8.8Hz,2H),5.98(d,J=10.0Hz,1H),3.97(d,J=11.6Hz,1H),3.74-3.68(m,1H),2.05(d,J=12.4 Hz, 1H), 1.93(d, J=12.4Hz, 1H), 1.77(d, J=8.0Hz, 1H), 1.59(s, 3H).ESI-MS m/z: 533[M+H]⁺ .

实施例22：Example 22:

1-环己基-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲1-cyclohexyl-3-[4-[[1-(2-tetrahydropyran)-1H-pyrazolo[3,4-d]pyrimidine]-4-oxyl]phenyl]urea

参照实施例4的制备方法。收率71.5％。1H NMR(400MHz,DMSO-d6)δ8.57(s,1H),8.46(s,1H),8.06(s,1H),7.47(d,J＝8.0Hz,2H),7.18(d,J＝8.0Hz,2H),6.13(d,J＝8.4Hz,1H),5.98(d,J＝9.2Hz,1H),5.59(d,J＝7.2Hz,4H),4.07(s,1H),3.96(d,J＝14.0Hz,1H),3.71(s,1H),2.00(s,1H),1.91(d,J＝14.0Hz,1H),1.80(s,2H),1.72(d,J＝12.0Hz,8H).ESI-MS m/z:437[M+H]⁺。Referring to the preparation method of Example 4. Yield 71.5%. 1H NMR (400MHz, DMSO-d6) δ8.57(s, 1H), 8.46(s, 1H), 8.06(s, 1H), 7.47(d, J=8.0Hz, 2H), 7.18(d, J= 8.0Hz, 2H), 6.13(d, J=8.4Hz, 1H), 5.98(d, J=9.2Hz, 1H), 5.59(d, J=7.2Hz, 4H), 4.07(s, 1H), 3.96 (d,J=14.0Hz,1H),3.71(s,1H),2.00(s,1H),1.91(d,J=14.0Hz,1H),1.80(s,2H),1.72(d,J= 12.0Hz, 8H). ESI-MS m/z: 437[M+H]⁺ .

实施例23：Example 23:

1-(3-甲基苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]硫脲1-(3-methylphenyl)-3-[4-[[1-(2-tetrahydropyran)-1H-pyrazolo[3,4-d]pyrimidine]-4-oxyl]benzene base] thiourea

参照实施例4的制备方法。收率69.5％。1H NMR(400MHz,DMSO-d6)δ9.86(d,J＝7.6Hz,2H),8.60(s,1H),8.15(s,1H),7.60(d,J＝8.8Hz,2H),7.31(d,J＝8.0Hz,4H),7.24(t,J＝7.6Hz,1H),6.97(d,J＝10.8Hz,1H),5.99(d,J＝9.6Hz,1H),3.97(d,J＝11.2Hz,1H),3.74–3.68(m,1H),2.31(s,3H),2.09(s,6H).13C NMR(101MHz,DMSO-d6)δ180.12,163.54,155.68,148.71,139.58,138.20,138.05,137.94,132.31,128.77,128.65,125.65,124.55,122.35,122.23,121.25,102.70,82.65,67.57,31.10,29.14,25.01,22.60,21.47.ESI-MS m/z:461[M+H]⁺。Referring to the preparation method of Example 4. Yield 69.5%. 1H NMR (400MHz, DMSO-d6) δ9.86 (d, J = 7.6Hz, 2H), 8.60 (s, 1H), 8.15 (s, 1H), 7.60 (d, J = 8.8Hz, 2H), 7.31 (d, J=8.0Hz, 4H), 7.24(t, J=7.6Hz, 1H), 6.97(d, J=10.8Hz, 1H), 5.99(d, J=9.6Hz, 1H), 3.97(d ,J＝11.2Hz,1H),3.74–3.68(m,1H),2.31(s,3H),2.09(s,6H).13C NMR(101MHz,DMSO-d6)δ180.12,163.54,155.68,148.71,139.58 ,138.20,138.05,137.94,132.31,128.77,128.65,125.65,124.55,122.35,122.23,121.25,102.70,82.65,67.57,31.10,29.14,25.01+MES-241.47 H]⁺ .

实施例24：Example 24:

1-(4-氯苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]硫脲1-(4-Chlorophenyl)-3-[4-[[1-(2-tetrahydropyran)-1H-pyrazolo[3,4-d]pyrimidine]-4-oxyl]phenyl ] Thiourea

参照实施例4的制备方法。收率为57.9％。1HNMR(400MHz，DMSO-d6)δ9.97(s，1H)，9.95(s，1H)，8.60(s，1H)，8.17(s，1H)，7.59(d，J＝8.8Hz，2H)，7.55(d，J＝8.8Hz，2H)，7.41(m，2H)，7.32(d，J＝8.8Hz，2H)，5.99(dd，J＝2.0Hz，2.4Hz，1H),3.97(d，J＝11.6Hz，1H)，3.72(m，1H)，2.46(m，1H)，2.05(d，J＝12.8Hz，1H)，1.93(dd，J＝2.8Hz，2.8Hz，1H)，1.77(m，1H)，1.60(m，2H).ESI-MS m/z:481[M+H]⁺。Referring to the preparation method of Example 4. The yield was 57.9%. 1HNMR (400MHz, DMSO-d6) δ9.97(s, 1H), 9.95(s, 1H), 8.60(s, 1H), 8.17(s, 1H), 7.59(d, J=8.8Hz, 2H), 7.55(d, J=8.8Hz, 2H), 7.41(m, 2H), 7.32(d, J=8.8Hz, 2H), 5.99(dd, J=2.0Hz, 2.4Hz, 1H), 3.97(d, J=11.6Hz, 1H), 3.72(m, 1H), 2.46(m, 1H), 2.05(d, J=12.8Hz, 1H), 1.93(dd, J=2.8Hz, 2.8Hz, 1H), 1.77 (m, 1H), 1.60 (m, 2H). ESI-MS m/z: 481 [M+H]⁺ .

实施例25：Example 25:

1-(3-三氟甲基-4-氯苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]硫脲1-(3-Trifluoromethyl-4-chlorophenyl)-3-[4-[[1-(2-tetrahydropyran)-1H-pyrazolo[3,4-d]pyrimidine]- 4-Oxy]phenyl]thiourea

参照实施例4的制备方法。收率为60.3％。1HNMR(400MHz，DMSO-d6)δ9.22(s，1H)，8.99(s，1H)，8.58(s，1H)，8.13(d，J＝3.2Hz，2H)，7.68(dd，J＝2.0Hz，2.0Hz，1H)，7.62(d，J＝8.8Hz，1H)，7.59(d，J＝8.8Hz，2H)，7.28(d，J＝9.2Hz，2H)，5.99(dd，J＝2.0Hz，2.4Hz，1H)，3.97(d，J＝11.2Hz，1H)，3.71(m，1H)，2.46(m，1H)，2.05(m，1H)，1.93(m，1H)，1.77(m，1H)，1.60(dt，2H).ESI-MS m/z:549[M+H]⁺。Referring to the preparation method of Example 4. The yield was 60.3%. 1HNMR (400MHz, DMSO-d6) δ9.22(s, 1H), 8.99(s, 1H), 8.58(s, 1H), 8.13(d, J=3.2Hz, 2H), 7.68(dd, J=2.0 Hz, 2.0Hz, 1H), 7.62(d, J=8.8Hz, 1H), 7.59(d, J=8.8Hz, 2H), 7.28(d, J=9.2Hz, 2H), 5.99(dd, J= 2.0Hz, 2.4Hz, 1H), 3.97(d, J=11.2Hz, 1H), 3.71(m, 1H), 2.46(m, 1H), 2.05(m, 1H), 1.93(m, 1H), 1.77 (m, 1H), 1.60 (dt, 2H). ESI-MS m/z: 549 [M+H]⁺ .

实施例26：Example 26:

1-(3-三氟甲基苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]硫脲1-(3-Trifluoromethylphenyl)-3-[4-[[1-(2-tetrahydropyran)-1H-pyrazolo[3,4-d]pyrimidine]-4-oxyl ]phenyl]thiourea

参照实施例4的制备方法。收率为69.9％。1HNMR(400MHz，DMSO-d6)δ10.12(s，1H)，10.11(s，1H)，8.60(s，1H)，8.19(s，1H)，7.99(s，1H)，7.79(d，J＝8.4Hz，1H)，7.59(q，J＝8.8Hz，7.2Hz，3H)，7.49(d，J＝7.6Hz，1H)，7.34(d，J＝8.8Hz，2H)，6.00(dd，J＝2.0Hz，2.4Hz，1H),3.97(d，J＝11.2Hz，1H)，3.72(m，1H)，2.46(dd，J＝4Hz，4Hz，1H)，2.05(d，J＝13.6Hz，1H)，1.93(dd，J＝2.8Hz，2.8Hz，1H)，1.79(m，1H)，1.61(m，2H).ESI-MS m/z:515[M+H]⁺。Referring to the preparation method of Example 4. The yield is 69.9%. 1H NMR (400MHz, DMSO-d6) δ10.12(s, 1H), 10.11(s, 1H), 8.60(s, 1H), 8.19(s, 1H), 7.99(s, 1H), 7.79(d, J =8.4Hz, 1H), 7.59(q, J=8.8Hz, 7.2Hz, 3H), 7.49(d, J=7.6Hz, 1H), 7.34(d, J=8.8Hz, 2H), 6.00(dd, J=2.0Hz, 2.4Hz, 1H), 3.97(d, J=11.2Hz, 1H), 3.72(m, 1H), 2.46(dd, J=4Hz, 4Hz, 1H), 2.05(d, J=13.6 Hz, 1H), 1.93 (dd, J = 2.8Hz, 2.8Hz, 1H), 1.79 (m, 1H), 1.61 (m, 2H). ESI-MS m/z: 515[M+H]⁺ .

实施例27：Example 27:

体外细胞株抑制活性(IC₅₀)测试In vitro cell line inhibitory activity (IC₅₀ ) test

采用MTT法测定化合物对高表达VEGFR-2的人脐静脉内皮细胞HUVEC，高表达BRAF(野生型)的人前列腺癌细胞PC-3以及高表达BRAF^V600E的人恶性黑色素瘤细胞A375、人结肠癌细胞HT-29、人前列腺癌细胞PC-3、人肺癌细胞A549四种肿瘤细胞株的体外抑制活性，并设定正常细胞犬肾细胞MDCK测定化合物的细胞毒性。MTT method was used to determine the effect of compounds on human umbilical vein endothelial cells HUVEC with high expression of VEGFR-2, human prostate cancer cell PC-3 with high expression of BRAF (wild type), human malignant melanoma cells A375 and human colon cancer cells with high expression of BRAF^V600E Cell HT-29, human prostate cancer cell PC-3, human lung cancer cell A549 four kinds of tumor cell lines in vitro inhibitory activity, and set normal cell canine kidney cell MDCK to determine the cytotoxicity of the compound.

体外培养细胞株，把对数生长期的六种细胞株细胞消化后，吹打成单细胞悬液，分别接种于96孔培养板；每孔5×10³个细胞，每孔加入培养基200μL，37℃、5％CO₂培养箱中培养过夜。待细胞贴壁后，分别加入不同剂量的受试化合物和阳性对照药索拉菲尼，配置不同浓度的样品，以空白组为阴性对照组，以索拉菲尼为阳性对照组，在培养箱中再培养48h。然后，每孔加入20μL质量浓度为5mg/mL的MTT液，连续培养4h。吸去上清液，每孔加入150μL二甲亚砜，将培养板置于微孔板振荡器上振荡10min，使结晶物溶解。在570nm波长处用酶标仪测吸光度A值，计算抑制率；按Bliss法计算IC₅₀。体外细胞活性测试的结果见下表1。Cultivate cell lines in vitro, digest six kinds of cell lines in the logarithmic growth phase, pipette into single cell suspension, and inoculate them in 96-well culture plates; 5×10³ cells per well, add 200 μL of medium to each well , and cultured overnight in a 37°C, 5% CO₂ incubator. After the cells adhered to the wall, different doses of the test compound and the positive control drug Sorafenib were added, and samples of different concentrations were prepared. The blank group was used as the negative control group, and Sorafenib was used as the positive control group. Incubate for another 48 hours. Then, 20 μL of MTT solution with a mass concentration of 5 mg/mL was added to each well, and cultured continuously for 4 h. Aspirate the supernatant, add 150 μL dimethyl sulfoxide to each well, and shake the culture plate on a microplate shaker for 10 min to dissolve the crystals. Measure the absorbance A value with a microplate reader at a wavelength of 570nm, and calculate the inhibition rate; calculate the IC₅₀ according to the Bliss method. The results of the in vitro cell viability test are shown in Table 1 below.

表1化合物对肿瘤细胞增殖结果Table 1 compounds on tumor cell proliferation results

体外细胞活性测试表明，化合物对高表达VEGFR-2、BRAF和BRAF^V600E的细胞具有较强的抑制活性，可用于预防或治疗与VEGFR-2、BRAF和BRAF^V600E有关的临床疾病，这些疾病可以是黑色素瘤、肝癌、肾癌、肺癌、骨癌、胰腺癌、皮肤癌、头颈癌、皮肤或眼内黑素瘤、子宫癌、卵巢癌、直肠癌、肛门区癌、胃癌、结肠癌、乳腺癌、输卵管癌、子宫内膜癌、宫颈癌、阴道癌、阴户癌、何杰金病、食道癌、小肠癌、内分泌系统癌、甲状腺癌、甲状旁腺癌、软组织肉瘤、尿道癌、阴茎癌、前列腺癌、慢性或急性白血病、膀胱癌、肾或输尿管癌、脊柱轴肿瘤、垂体腺瘤、胃肠间质肿瘤、结肠直肠癌、非小细胞肺癌、小细胞肺癌、肥大细胞增多症、胶质瘤、肉瘤、淋巴瘤等。The in vitro cell activity test shows that the compound has strong inhibitory activity on cells with high expression of VEGFR-2, BRAF and BRAF^V600E , and can be used to prevent or treat clinical diseases related to VEGFR-2, BRAF and BRAF^V600E . These diseases can be Melanoma, liver cancer, kidney cancer, lung cancer, bone cancer, pancreatic cancer, skin cancer, head and neck cancer, skin or intraocular melanoma, uterine cancer, ovarian cancer, rectal cancer, anal region cancer, stomach cancer, colon cancer, breast cancer , fallopian tube cancer, endometrial cancer, cervical cancer, vaginal cancer, vulvar cancer, Hodgkin's disease, esophagus cancer, small intestine cancer, endocrine system cancer, thyroid cancer, parathyroid cancer, soft tissue sarcoma, urethral cancer, penile cancer, Prostate cancer, chronic or acute leukemia, bladder cancer, kidney or ureter cancer, spinal axis tumors, pituitary adenomas, gastrointestinal stromal tumors, colorectal cancer, non-small cell lung cancer, small cell lung cancer, mastocytosis, glial Tumors, sarcomas, lymphomas, etc.

Claims (8)

Translated fromChinese

1.一种具有抗肿瘤活性的1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶类化合物，其特征在于：该化合物选自1-(3-甲基苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲、1-(3,4-二氯苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲、1-(2-氯-5-甲基苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲、1-(2,3-二甲基苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲、1-(4-氟-3-甲基苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲、1-(2-氟-5 -(三氟甲基)苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲、1-(3-氰基苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲、1-(5-甲基-2-硝基苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲、1-(2-甲基苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲、1-(3-氟苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲、1-(2,5-二甲基苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲、1-(3-三氟甲基苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲、1-(3-三氟甲基-4-氯苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲、1-(3-氟-5-三氟甲基苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲、1-(4-氯苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲、1-(2-氯-5-三氟甲基苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲、1-环己基-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]脲、1-(4-氯苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]硫脲、1-(3-三氟甲基-4-氯苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]硫脲、1-(3-三氟甲基苯基)-3-[4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯基]硫脲中的其中一种。1. A 1-(2-tetrahydropyran)-1H -pyrazolo[3,4-d]pyrimidine compound with antitumor activity, characterized in that: the compound is selected from 1-(3- methylphenyl)-3-[4-[[1-(2-tetrahydropyran)-1H -pyrazolo[3,4-d]pyrimidine]-4-oxyl]phenyl]urea, 1-(3,4-Dichlorophenyl)-3-[4-[[1-(2-tetrahydropyran)-1H -pyrazolo[3,4-d]pyrimidine]-4-oxo base]phenyl]urea, 1-(2-chloro-5-methylphenyl)-3-[4-[[1-(2-tetrahydropyran)-1H -pyrazolo[3,4 -d]pyrimidine]-4-oxyl]phenyl]urea, 1-(2,3-dimethylphenyl)-3-[4-[[1-(2-tetrahydropyran)-1H -pyrazolo[3,4-d]pyrimidine]-4-oxyl]phenyl]urea, 1-(4-fluoro-3-methylphenyl)-3-[4-[[1-(2 -tetrahydropyran)-1H -pyrazolo[3,4-d]pyrimidine]-4-oxyl]phenyl]urea, 1-(2-fluoro-5-(trifluoromethyl)phenyl )-3-[4-[[1-(2-tetrahydropyran)-1H -pyrazolo[3,4-d]pyrimidine]-4-oxyl]phenyl]urea, 1-(3 -cyanophenyl)-3-[4-[[1-(2-tetrahydropyran)-1H -pyrazolo[3,4-d]pyrimidine]-4-oxyl]phenyl]urea , 1-(5-methyl-2-nitrophenyl)-3-[4-[[1-(2-tetrahydropyran)-1H -pyrazolo[3,4-d]pyrimidine] -4-oxyl]phenyl]urea, 1-(2-methylphenyl)-3-[4-[[1-(2-tetrahydropyran)-1H -pyrazolo[3,4 -d]pyrimidine]-4-oxyl]phenyl]urea, 1-(3-fluorophenyl)-3-[4-[[1-(2-tetrahydropyran)-1H -pyrazolo [3,4-d]pyrimidine]-4-oxyl]phenyl]urea, 1-(2,5-dimethylphenyl)-3-[4-[[1-(2-tetrahydropyran )-1H -pyrazolo[3,4-d]pyrimidine]-4-oxyl]phenyl]urea, 1-(3-trifluoromethylphenyl)-3-[4-[[1- (2-tetrahydropyran)-1H -pyrazolo[3,4-d]pyrimidine]-4-oxyl]phenyl]urea, 1-(3-trifluoromethyl-4-chlorophenyl )-3-[4-[[1-(2-tetrahydropyran)-1H -pyrazolo[3,4-d]pyrimidine]-4-oxyl]phenyl]urea, 1-(3 -Fluoro-5-trifluoromethylphenyl)-3-[4-[[1-(2-tetrahydropyran)-1H -pyrazolo[3,4-d]pyrimidine]-4-oxo base]phenyl]urea, 1-(4-chlorophenyl)-3-[4-[[1-(2-tetrahydropyran)-1H -pyrazolo[3,4-d]pyrimidine] -4-oxyl]phenyl]urea, 1-(2-chloro-5-trifluoromethylphenyl)-3-[4-[[1-(2-tetrahydro Pyran)-1H -pyrazolo[3,4-d]pyrimidine]-4-oxyl]phenyl]urea, 1-cyclohexyl-3-[4-[[1-(2-tetrahydropyridine Fyran)-1H -pyrazolo[3,4-d]pyrimidine]-4-oxyl]phenyl]urea, 1-(4-chlorophenyl)-3-[4-[[1-(2 -tetrahydropyran)-1H-pyrazolo[3,4-d ]pyrimidine]-4-oxyl]phenyl]thiourea, 1-(3-trifluoromethyl-4-chlorophenyl)- 3-[4-[[1-(2-tetrahydropyran)-1H -pyrazolo[3,4-d]pyrimidine]-4-oxyl]phenyl]thiourea, 1-(3- Trifluoromethylphenyl)-3-[4-[[1-(2-tetrahydropyran)-1H -pyrazolo[3,4-d]pyrimidine]-4-oxyl]phenyl] One of the thioureas.2.权利要求1中所述的一种具有抗肿瘤活性的1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶类化合物合成方法，其特征在于：将4,6-二氯-5-嘧啶甲醛与一水合肼进行环合反应生成4-氯-1H-吡唑并[3,4-d]嘧啶，在催化剂作用下，与3,4-二氢-2H-吡喃反应，生成4-氯-1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶，在保护气体保护下，与对氨基酚反应得到胺类中间体4-[[1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶]-4-氧基]苯胺，最后与异氰酸酯或硫异氰酸酯反应得到目标化合物。2. a kind of 1-(2-tetrahydropyran)-1H -pyrazolo[3,4-d] pyrimidine synthetic method with antitumor activity described in claim 1 is characterized in that: 4,6-dichloro-5-pyrimidine formaldehyde and hydrazine monohydrate are subjected to ring closure reaction to generate 4-chloro-1H -pyrazolo[3,4-d]pyrimidine, under the action of catalyst, with 3,4- Dihydro-2H-pyran reaction, generate 4-chloro-1-(2-tetrahydropyran)-1H -pyrazolo[3,4-d]pyrimidine, under the protection of protective gas, with p-aminophenol The reaction gives the amine intermediate 4-[[1-(2-tetrahydropyran)-1H -pyrazolo[3,4-d]pyrimidine]-4-oxyl]aniline, and finally with isocyanate or thioisocyanate The reaction yields the target compound.3.根据权利要求2中所述的一种具有抗肿瘤活性的1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶类化合物合成方法，其特征在于：所述的催化剂为酸性催化剂和盐类催化剂中的至少一种。3. according to a kind of 1-(2-tetrahydropyran)-1H -pyrazolo[3,4-d] pyrimidine synthetic method with antitumor activity described in claim 2, it is characterized in that : the catalyst is at least one of an acid catalyst and a salt catalyst.4.根据权利要求3中所述的一种具有抗肿瘤活性的1-(2-四氢吡喃)-1H-吡唑并[3,4-d]嘧啶类化合物合成方法，其特征在于：所述的保护气体为氮气和惰性气体中的至少一种。4. according to a kind of 1-(2-tetrahydropyran)-1H -pyrazolo[3,4-d] pyrimidine synthetic method with antitumor activity described in claim 3, it is characterized in that : The protective gas is at least one of nitrogen and inert gas.5.一种药物组合物，其特征在于：包括下列成分中的至少一种：a）化合物、b）该化合物在药学上可接受的盐；其中，所述的化合物为权利要求1所述的化合物。5. A pharmaceutical composition, characterized in that it includes at least one of the following components: a) a compound, b) a pharmaceutically acceptable salt of the compound; wherein the compound is the compound described in claim 1 compound.6.权利要求5所述的一种药物组合物在制备因蛋白激酶活性异常所引起疾病的药物的应用。6. The application of a pharmaceutical composition according to claim 5 in the preparation of medicines for diseases caused by abnormal protein kinase activity.7.根据权利要求6中所述的应用，其特征在于：所述的疾病为癌症。7. The use according to claim 6, characterized in that: said disease is cancer.8.根据权利要求7中所述的应用，其特征在于：所述的疾病为黑色素瘤、肝癌、肾癌、肺癌、骨癌、胰腺癌、皮肤癌、头颈癌、子宫癌、卵巢癌、直肠癌、肛门区癌、胃癌、结肠癌、乳腺癌、输卵管癌、宫颈癌、阴道癌、阴户癌、何杰金病、食道癌、小肠癌、内分泌系统癌、甲状腺癌、甲状旁腺癌、软组织肉瘤、尿道癌、阴茎癌、前列腺癌、慢性或急性白血病、膀胱癌、脊柱轴肿瘤、垂体腺瘤、胃肠间质肿瘤、结肠直肠癌、肥大细胞增多症、胶质瘤、肉瘤、淋巴瘤中的一种或任意几种的组合。8. The application according to claim 7, characterized in that: the disease is melanoma, liver cancer, kidney cancer, lung cancer, bone cancer, pancreatic cancer, skin cancer, head and neck cancer, uterine cancer, ovarian cancer, rectal cancer Cancer, anal region cancer, stomach cancer, colon cancer, breast cancer, fallopian tube cancer, cervical cancer, vaginal cancer, vulvar cancer, Hodgkin's disease, esophagus cancer, small intestine cancer, endocrine system cancer, thyroid cancer, parathyroid cancer, soft tissue Sarcoma, urethral cancer, penile cancer, prostate cancer, chronic or acute leukemia, bladder cancer, spinal axis tumors, pituitary adenomas, gastrointestinal stromal tumors, colorectal cancer, mastocytosis, glioma, sarcoma, lymphoma one or any combination of several.