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WO2018068665A1 - 微管蛋白抑制剂 - Google Patents

微管蛋白抑制剂 Download PDF

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WO2018068665A1
WO2018068665A1 PCT/CN2017/104307 CN2017104307W WO2018068665A1 WO 2018068665 A1 WO2018068665 A1 WO 2018068665A1 CN 2017104307 W CN2017104307 W CN 2017104307W WO 2018068665 A1 WO2018068665 A1 WO 2018068665A1
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propenylene
group
compound
piperazine
tert
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PCT/CN2017/104307
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English (en)
French (fr)
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唐田
彭江华
吴婧
冯贻东
杨经安
佘琴
冯汉林
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深圳海王医药科技研究院有限公司
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Publication of WO2018068665A1 publication Critical patent/WO2018068665A1/zh

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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
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/337—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having four-membered rings, e.g. taxol
    • 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
    • A61K31/496—Non-condensed piperazines containing further heterocyclic rings, e.g. rifampin, thiothixene or sparfloxacin
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
    • C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
    • C07D403/06—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings linked by a carbon chain containing only aliphatic carbon atoms

Definitions

  • the present invention relates to the field of medicine, and in particular to a novel series of compounds as tubulin inhibitors and uses thereof.
  • the invention also relates to intermediate compounds for the preparation of said compounds.
  • the invention further relates to a pharmaceutical composition comprising the compound.
  • microtubule inhibitors drugs acting on microtubules
  • the current clinical drugs are affected by the following unfavorable problems: poor water solubility, unfavorable allergic reactions, serious toxic side effects and acquired drug resistance, resulting in reduced efficacy, complicated chemical structure and difficulty in synthesis.
  • the scarcity of sources limits their further use. Therefore, there is an urgent need to find new types of tubulin inhibitors, especially small molecule inhibitors with simple structures.
  • Microtubules are important components of eukaryotic cells and important targets for antitumor drugs. Microtubules are the main components of the cytoskeleton. They are composed of ⁇ -tubulin and ⁇ -tubulin heterodimer and have the characteristics of hollow tubular structure. In addition, there is a gamma tubulin, which is not a component of microtubules but participates in the assembly of microtubules.
  • Microtubules have the dynamic properties of polymerization and depolymerization, and play an important role in cell morphology, cell division, signal transduction and material transport. Microtubules polymerize into spindles in the early stage of cell division, and the spindle in the mitosis pulls the chromosomes into the two poles and moves into the two daughter cells to complete cell proliferation. Since microtubules play an extremely important role in cell division, they have become one of the important targets for antitumor drug research. Tubulin inhibitors acting on microtubule systems have also become an effective class of antitumor drugs.
  • tubulin inhibitors There are two classification methods for tubulin inhibitors. One is divided into two types according to the different mechanism of action: 1 tubulin depolymerization inhibitor that inhibits tubulin polymerization; 2 tubulin polymerization agent that promotes tubulin polymerization. Another classification method is divided into three categories according to the different sites of tubulin inhibitor and tubulin: 1 tubulin inhibitor acting on the colchicine site; 2 microactin acting on the vinblastine site Tubulin inhibitor; 3 tubulin inhibitor acting on the paclitaxel site.
  • the colchicine site is conducive to the study of small molecule anti-tumor inhibitors due to the small volume of its own binding cavity.
  • Chinese patent application CN 1684955 relates to a compound for the treatment of cancer and fungal infections, dehydrophenyl phenyl pinabulin (formula II), which is a cell cycle inhibitor, as a tumor growth inhibitor or Fungal inhibitors.
  • Chinese patent application CN1934101A relates to the use of the above compounds for reducing vascular proliferation and vascular density, acting on tumor blood vessels, but plinabulin alone has a weaker inhibitory effect on tumors and a greater interference with the immune system.
  • the properties, key functional residues and potential binding sites of each sub-region are determined; starting from the common group in the inhibitor structure, the consensus is gradually expanded.
  • the nature of the group thus assuming a structural template for the inhibitor.
  • the three-dimensional pharmacophore model of the inhibitor and some structural factors affecting the activity were proposed.
  • the design, synthesis, structural modification and in vitro activity studies of novel microtubule inhibitors were carried out.
  • Another object of the present invention is to provide an intermediate for the preparation of the above compounds.
  • the invention also provides a pharmaceutical composition comprising the above-described tubulin inhibitor.
  • tubulin inhibitor having the structure of formula (I):
  • n independently represents an integer of 0 to 5, with the proviso that n ⁇ 5, and A represents a mono- or poly-substituted group selected from H, C 1 -C 20 alkyl, C 1 -C 20 hydrocarbyl, C 1- C 20 acylamino group, C 1 -C 20 acyloxy group, C 1 -C 20 alkanoyl group, C 1 -C 20 alkoxycarbonyl group, C 1 -C 20 alkoxy group, C 1 -C 20 alkylamino group, C 1 -C 20 alkylcarboxyamino, aroyl, aralkyl, carboxyl, cyano, halogen, hydroxy, nitro and methylthienyl.
  • n independently represents an integer of 0 to 2, with the proviso that n ⁇ 2, and A represents a mono- or poly-substituted group selected from the group consisting of H, vinyl, and A.
  • Base trifluoromethoxy, methoxy, cyano, halogen and benzoyl.
  • Representative compounds of the invention include:
  • the synthetic route 1 of the present invention is as follows:
  • a pharmaceutical composition comprising a therapeutically effective amount of a compound of formula (I) and a pharmaceutically acceptable carrier and/or adjuvant for the treatment of various cancers, infections, inflammations and conventional proliferation Sexual disease, or treatment of other diseases characterized by the appearance of rapidly proliferating cells such as psoriasis and other skin diseases.
  • the therapeutically effective amount means that the amount of the compound of formula (I) contained in the pharmaceutical composition is sufficient to produce a clinically desired therapeutic effect, e.g., the tumor size of the patient is reduced to a clinically acceptable range.
  • the compounds or pharmaceutical compositions of the invention are used to treat sarcomas, carcinomas and/or leukemias.
  • exemplary conditions in which the compound or composition can be used alone or as part of a therapeutic regimen include fibrosarcoma, mucinous sarcoma, liposarcoma, chondrosarcoma, osteogenic sarcoma, chordoma, angiosarcoma, endothelial sarcoma, lymphangiosarcoma , lymphatic endothelial sarcoma, synovial tumor, mesothelioma, Ewing's tumor, leiomyomas, rhabdomyosarcoma, colon cancer, pancreatic cancer, breast cancer, ovarian cancer, prostate cancer, squamous cell carcinoma, basal cell carcinoma, sweat gland Cancer, sebaceous gland cancer, papillary carcinoma, papillary adenocarcinoma, cystadenocarcinoma, medullary carcinoma,
  • the compounds or compositions of the invention are used to treat conditions such as cancer of the breast, prostate, kidney, bladder, or colon tissue.
  • the compounds or pharmaceutical compositions of the invention are used to treat proliferative or neoplastic diseases occurring in adipose tissue, such as adipocyte tumors, namely lipomas, fibroids, lipoblastoma, fat Symptomatic, brown lipoma (hibemomas), hemangiomas and/or liposarcoma.
  • adipocyte tumors namely lipomas, fibroids, lipoblastoma, fat Symptomatic, brown lipoma (hibemomas), hemangiomas and/or liposarcoma.
  • compositions and compounds can also be used to control infectious agents and parasites (e.g., bacteria, trypanosomes, fungi, etc.).
  • infectious agents and parasites e.g., bacteria, trypanosomes, fungi, etc.
  • the pharmaceutical composition of the present invention can be administered by intravenous, intradermal, intramuscular, subcutaneous, oral or inhalation, and the pharmaceutical composition can be used as a gastrointestinal preparation such as a tablet, a capsule or a pill. Alternatively, it may be a parenteral preparation such as an injection or a topical preparation.
  • the invention provides a method of treating an anti-tumor and related disease, the method comprising administering to a patient having cancer or a related condition a therapeutically effective amount of a compound of the formula (I).
  • the invention also provides a method of modulating microtubule polymerization in a patient, the method comprising administering a therapeutically effective amount of at least one compound of the invention or a pharmaceutically acceptable prodrug, salt, hydrate, solvate, crystalline form thereof Or diastereomers.
  • the compounds of the present invention can be used in combination with chemotherapeutic agents for the treatment of tumors, and experiments have shown that the compounds of the present invention can increase the tumor suppressing effect when combined with a chemotherapeutic agent, while reducing the toxicity of the chemotherapeutic agent.
  • the most preferred compound is compound (9), and the chemotherapeutic agent can be a conventional chemotherapeutic agent generally used for treating tumors.
  • compound (9) is used in combination with docetaxel to obtain a very good therapeutic effect. .
  • the compound of the invention can bind to tubulin, can inhibit the growth and proliferation of cancer cells, has almost no influence on the corresponding white blood cells and granulocytes, and has less interference with immune system function when fighting tumor proliferation, and greatly improves anti-tumor drugs.
  • the drawbacks of clinical use Such compounds are also useful in the treatment of other conditions associated with hyperproliferation.
  • Figure 1 shows the tumor growth inhibition effect of the compound of the present invention
  • Figure 2 shows the antitumor effect and safety test results of compound (9).
  • A Inhibition of tumor growth inhibition in nude mice
  • B Effect of compound on tumor weight in nude mice
  • C Compound (9) and continuous administration of docetaxel for SD The effect of murine neutrophil count
  • Figure 3 is a graph showing the effect of the compounds of the invention on neutrophil counts in rats.
  • the structural modification of the compound of the present invention is based on the natural product Phenylahistin as a lead compound.
  • the corresponding structural transformation is carried out in combination with the reported structure-activity relationship. According to the principle of olefin insertion, a double bond is inserted between the piperazine and the benzene ring.
  • the piperazine diene and the benzene ring are directly connected, and different substituents are introduced to the benzene ring, it may be A compound having similar activity or more activity is obtained; and a double olefin bond is further inserted between the piperazine ring and the pyrazole to make the compound more stable and not easily deteriorated at normal temperature, so that the compound of the present invention has better stability and safety. high.
  • TLC was carried out on a silica gel GF254 high efficiency plate (Yantai Zhifu Silica Gel Development Test Plant).
  • the optical rotation measurement was carried out on a WZZ-1S optical rotation measuring instrument (Shanghai Precision Scientific Instrument Co., Ltd.).
  • step 1
  • the intermediate B was prepared in the same manner as in Steps 1 and 2, and the same as Steps 1 and 2 in Example 1.
  • the preparation of other intermediates and final products is shown in the following table:
  • HTRF kinEASE TK kit was used to detect the inhibitory activity of 11 compounds on Lyn ⁇ , AKT2, KDR, IKK- ⁇ , c-RAF enzyme, and the test compound was diluted from 10 ⁇ L to 0.1 ⁇ M to control the DMSO concentration. 1%, each concentration is a duplicate well.
  • HT-1080CAKi-1 human fibrosarcoma cells AGS human gastric adenocarcinoma cells, A375 human skin melanoma cells, Fadu human pharyngeal squamous carcinoma cells, NCI-H1975 human lung adenocarcinoma cells, SK-ES-1 human osteosarcoma cells, MDA- MB-231 human breast cancer cells and MIAPaCa-2 human pancreatic cancer cells were all derived from ATCC.
  • the cells were digested from the cell culture plate using trypsin, and the cell density was measured after resuspending in the medium.
  • McCoys'5A medium is used to culture SK-ES-1 cells (INVITROGEN)
  • Luminescent cell viability assay kit (promega#G7572)
  • the cell culture plate to which the compound was added was returned to the incubator and incubated for 72 hours under humidified conditions of 37 ° C, 5% CO 2 .
  • the luminescent cell viability assay kit (promega #G7572), the substrate lyophilized powder and the buffer were mixed, added to a 96-well plate at a dose of 30 ul/well, and placed at room temperature for 30 minutes before the experiment to equilibrate.
  • the time to read the board is set to 0.5 seconds per hole.
  • the above compounds had a strong inhibitory effect on each of the tumor cell lines examined, and compared with the reference drug [(-)phenyl Astrastin] (commercially available), the compound in this example inhibited the tumor with a high intensity.
  • the concentration of the compound in the present embodiment to inhibit the tumor is significantly lower than the concentration of the inhibitory enzyme, so it can be inferred that the compound of the present embodiment inhibits tumor proliferation.
  • it has a strong inhibitory effect on other enzyme activities or intracellular molecules that have not been investigated, and may affect various intracellular and extracellular targets to exert strong synergistic inhibition to varying degrees. Tumor effect.
  • MES-SA human uterine sarcoma cells (derived from ATCC) were cultured in low concentrations of doxorubicin for a long time to maintain sufficient cell viability and induce the expression of p-glycoprotein. After 10 months of culture, the expression results were induced by WB detection. After successful induction, after one week of culture without doxorubicin medium, the cell concentration was adjusted, and the tumor cells were inoculated into the armpit of nude mice, and the transplanted tumors were proliferated to a certain volume and then treated with different drugs. The doses of different doses of different drugs were used to explore the doses that did not affect the body weight gain of the animals, which was the dose to be treated for the transplanted tumor animals (Fig. 1).
  • Tumor inhibition rate (%) (1 - treatment group tumor weight / control tumor weight) * 100%, according to Figure 1, the tumor inhibition rate of docetaxel is close to 80%, and higher than compound 9, compound 11 , Compound 15, Compound 18.
  • the nude mice inoculated with tumor cells were given blank vehicle, docetaxel, compound 9 and docetaxel and compound 9 in combination, and the tumor volume was measured at different time points after drug administration and the tumor volume change process was detected. At the same time, the change in body weight of nude mice was examined to evaluate the antitumor effect and the degree of toxicity of the drug. In addition, in normal SD rats, blood neutrophil counts were measured before and after the administration according to the above administration and grouping methods to evaluate the degree of influence of the drug on neutrophil counts.
  • the multi-dose was given different drugs to inhibit the degree of tumor xenografting in nude mice, and the equivalent dose of each drug was explored, and different drugs were given to rats by the inter-species dose conversion. That is, the effect of the drug on neutrophil count in rats was examined under the dose of the same pharmacodynamic dose to the rats.
  • Human umbilical vein endothelial cells (HuVEC from Cambrex) were used in this study to evaluate compounds 9, 11, 15, 18 and tert-butylphenyl by staining ⁇ -tubulin with colchicine and paclitaxel. The effect of ustin on tubulin.
  • the compounds according to the invention may be combined with a physiologically acceptable carrier or vehicle to provide a pharmaceutical composition, such as a lyophilized powder in the form of a tablet or capsule containing various fillers and binders.
  • a pharmaceutical composition such as a lyophilized powder in the form of a tablet or capsule containing various fillers and binders.
  • the compounds can be co-administered with other agents, and co-administration should mean at least two agents are used in the patient to provide the beneficial effects of the combination of the two agents.
  • the medicament can be used simultaneously or sequentially in a certain period of time.
  • the effective dosage of the compound in the composition can be determined extensively by one skilled in the art.
  • the compounds of the invention may be used alone or in combination with one or more additional agents.
  • Combination therapies contemplated by the present invention include, for example, the use of the compounds of the invention and additional agents in a single pharmaceutical formulation, as well as the use of the compounds of the invention and additional agents in separate pharmaceutical formulations.
  • the compounds of the invention may be administered to a dairy animal, preferably a human, alone or in combination with a pharmaceutically acceptable carrier or diluent, optionally together with known adjuvants such as microcrystalline cellulose, in a pharmaceutical composition according to standard pharmaceutical practice.
  • the compounds can be administered orally or parenterally, including intravenous, intramuscular, intraperitoneal, subcutaneous, rectal and topical routes of administration.
  • the selected compound can be administered, for example, in the form of a tablet or capsule or as an aqueous solution or suspension.
  • usual carriers are usually added including lactose and corn starch, and lubricants such as magnesium stearate.
  • diluents which may be employed include lactose and anhydrous corn starch, and when an aqueous suspension for oral use is required, the active ingredient is mixed with emulsifying and suspending agents. If desired, some sweeteners and/or flavoring agents may be added.
  • the total concentration of solutes should be controlled to make the formulation isotonic.
  • the active compound, the disintegrant and the filler of the present invention are sieved in a ratio of 60-100 mesh, uniformly mixed, and the soft material is prepared by using 2 to 20% of povidone K30 in ethanol solution, and granulating through 20 to 50 mesh. Dry at 40 ⁇ 90°C, the moisture content of the granules is controlled within 3%. After the whole granules, add appropriate amount of lubricant, mix well, and compress.
  • the pharmaceutical composition of the above embodiment can also be prepared by weighing the compound 9, 18, the filler and the disintegrant in a 50-fold prescription amount through a 60- and 80-mesh sieve, and mixing uniformly, using 2 to 20 % povidone K30 50% ethanol solution soft material, 30 mesh sieve granulation, 60 ° C drying, particle moisture control within 3%, 20 mesh sieve granules, add appropriate amount of lubricant, mix evenly, tablet, That is the product.
  • the obtained compound 9 was subjected to stability investigation (10-day accelerated test), and the moisture, purity, maximum single and total impurities, and 0 days of the compound were observed at 40 ° C, 60 ° C, humidity of 75%, 92.5%, and light conditions. The data were compared and the results showed that the obtained compound was stable. However, the degradation of (-) phenyl azanthine was obvious at 60 °C, indicating that high temperature has an effect on the stability of (-) phenyl axidine.

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Abstract

本发明的提供一种新的微管蛋白抑制剂及其应用，该新的微管蛋白抑制剂为一系列基于被取代的杂环骨架的化合物，以微管蛋白中的秋水仙碱结合位点为靶标。具有下述式(I)结构：其中：式(A)，n独立表示0～5的整数，条件是n≤5，A表示单取代或多取代的基团，所述基团选自H、C1-C20酰氨基、C1-C20酰氧基、C1-C20烷酰基、C1-C20烷氧羰基、C1-C20烷氧基、C1-C20烷氨基、C1-C20烷羧氨基、芳酰基、芳烷酰基、羧基、氰基、卤素、羟基、硝基和甲基噻吩基。

Description

微管蛋白抑制剂 技术领域
本发明涉及药物领域，具体地说涉及作为微管蛋白抑制剂的新的系列化合物及其应用。
本发明也涉及制备所述化合物的中间体化合物。
本发明还涉及含有该化合物的药物组合物。
背景技术
作为肿瘤治疗的主要手段，抗肿瘤药物为延长患者的生存时间以及改善其生命质量做出了相当的贡献。其中作用于微管的药物(微管抑制剂)，又在肿瘤药物中具有相当重要的地位。但是目前的临床药物，受到如下不良问题的影响：较差的水溶性，不利于给药且易造成的过敏反应、严重的毒副作用以及获得性耐药性，导致疗效降低、化学结构复杂难于合成而造成来源稀少，限制了它们进一步的使用。因而，急需寻找设计新型的微管蛋白抑制剂，尤其是结构简单的小分子抑制剂。
微管(microtubule)是真核细胞的重要组分，也是重要的抗肿瘤药物作用靶标。微管是细胞骨架的主要组成部分,由α-微管蛋白和β-微管蛋白异二聚体组成,具有中空管状结构的特点。此外,还有一种γ微管蛋白,它不是微管的组成成分,但参与微管的组装。
微管具有聚合和解聚的动力学特性，在细胞形态、细胞分裂、信号转导及物质输送等过程中起着重要作用。微管在细胞分裂前期聚合成为纺锤体,而纺锤体在有丝分裂中牵引染色体向两极移动进入两个子细胞中,完成细胞增殖。由于微管在细胞分裂中具有极其重要的作用,现已成为抗肿瘤药物研究的重要靶点之一。作用于微管系统的微管蛋白抑制剂也已成为一类有效的抗肿瘤药物。
微管蛋白抑制剂有两种分类方法。一种是根据作用机制的不同分为两种类型:①抑制微管蛋白聚合的微管蛋白解聚剂；②促进微管蛋白聚合的微管蛋白聚合剂。另一种分类方法是根据微管蛋白抑制剂与微管蛋白作用位点的不同分为3类:①作用于秋水仙碱位点的微管蛋白抑制剂；②作用于长春碱位点的微管蛋白抑制剂；③作用于紫杉醇位点的微管蛋白抑制剂。
现有研究表明，微管蛋白(tubulin)中存在三个主要的药物结合位点：紫杉醇结合位点(Taxol site)、长春碱结合位点(Vinblastine sitc)以及秋水仙碱结合位点(Colchieinesite)。但在这三个位点中，秋水仙碱位点由于自身结合腔的体积较小，有利于小分子抗肿瘤抑制剂的研究。
中国专利申请CN 1684955涉及一种治疗癌症和真菌感染的化合物脱氢苯基阿夕斯丁(plinabulin)(式II)，其为细胞周期抑制剂，作为肿瘤生长抑制剂或 真菌抑制剂。
Figure PCTCN2017104307-appb-000001
中国专利申请CN1934101A涉及上述化合物的用途，用于减少血管增生和血管密度，作用于肿瘤血管，但plinabulin单独使用对肿瘤的抑制作用较弱，对免疫系统的功能干扰较大。
发明内容
本发明的一个目的是提供一种新的微管蛋白抑制剂，为一系列基于被取代的杂环骨架的化合物，以微管蛋白中的秋水仙碱结合位点为靶标。
通过分析研究结合位点的结构性质，并模拟其与抑制剂的结合，确定各亚区域的性质、关键作用残基以及潜在的结合部位；从抑制剂结构中的共有基团入手，逐步扩充共有基团的性质，从而假设了抑制剂的结构模板。再从活性构象出发并结合已有的构效关系研究，提出抑制剂的三维药效团模型以及影响活性的部分结构因素。在此基础上，开展新型微管抑制剂的设计、合成、结构改造以及体外活性研究。
本发明的另一个目的是提供一种制备上述化合物的中间体。
本发明还提供包含上述微管蛋白抑制剂的药物组合物。
根据本发明的一方面，一种微管蛋白抑制剂，具有式(I)结构：
Figure PCTCN2017104307-appb-000002
其中：
Figure PCTCN2017104307-appb-000003
n独立表示0～5的整数，条件是n≤5，A表示单取代或多取代的基团，所述基团选自H、C1-C20烷基、C1-C20烃基、C1-C20酰氨基、C1-C20酰氧基、C1-C20烷酰基、C1-C20烷氧羰基、C1-C20烷氧基、C1-C20烷氨基、C1-C20烷羧氨基、芳酰基、芳烷酰基、羧基、氰基、卤素、羟基、硝基和甲基噻吩基。
更优选的是，上述式(I)中，n独立表示0～2的整数，条件是n≤2，A表示单取代或多取代的基团，所述基团选自H、乙烯基、甲基、三氟甲氧基、甲氧基、氰基、卤素和苯甲酰基。
本发明的代表性化合物包括：
(3Z,6Z)-3-((E)-3-(5-叔丁基)-1H-咪唑基-4-基)丙烯亚基)-6-((E)-3-(3-乙烯基苯基)-2-丙烯亚基)哌嗪-2,5-二酮(9)；
(3Z,6Z)-3-((E)-3-(5-叔丁基)-1H-咪唑基-4-基)丙烯亚基)-6-((E)-3-(3-氟苯基)-2-丙烯亚基)哌嗪-2,5-二酮(10)；
(3Z,6Z)-3-((E)-3-(5-叔丁基)-1H-咪唑基-4-基)丙烯亚基)-6-((E)-3-苯基丙烯亚基)哌嗪-2,5-二酮(11)；
(3Z,6Z)-3-((E)-3-(5-叔丁基)-1H-咪唑基-4-基)丙烯亚基)-6-((E)-3-(2,3-二甲基苯基)-2-丙烯亚基)哌嗪-2,5-二酮(12)；
(3Z,6Z)-3-((E)-3-(5-叔丁基)-1H-咪唑基-4-基)丙烯亚基)-6-((E)-3-(3-三氟甲氧基苯基)-2-丙烯亚基)哌嗪-2,5-二酮(13)；
(3Z,6Z)-3-((E)-3-(5-叔丁基)-1H-咪唑基-4-基)丙烯亚基)-6-((E)-3-(2,5-二氟苯基)-2-丙烯亚基)哌嗪-2,5-二酮(14)；
(3Z,6Z)-3-((E)-3-(5-叔丁基)-1H-咪唑基-4-基)丙烯亚基)-6-((E)-3-(3-甲氧基苯基)-2-丙烯亚基)哌嗪-2,5-二酮(15)；
(3Z,6Z)-3-((E)-3-(5-叔丁基)-1H-咪唑基-4-基)丙烯亚基)-6-((E)-3-(3,5-二甲氧基苯基)-2-丙烯亚基)哌嗪-2,5-二酮(16)；
(3Z,6Z)-3-((E)-3-(5-叔丁基)-1H-咪唑基-4-基)丙烯亚基)-6-((E)-3-(3-氯苯基)-2-丙烯亚基)哌嗪-2,5-二酮(17)；
(3Z,6Z)-3-((E)-3-(5-叔丁基)-1H-咪唑基-4-基)丙烯亚基)-6-((E)-3-(3-腈基苯基)-2-丙烯亚基)哌嗪-2,5-二酮(18)；
(3Z,6Z)-3-((E)-3-(5-叔丁基)-1H-咪唑基-4-基)丙烯亚基)-6-((E)-3-(3-苯甲酰基苯基)-2-丙烯亚基)哌嗪-2,5-二酮(19)；
根据本发明的另一方面，提供制备本发明化合物的中间体，具有式(B)结构：
Figure PCTCN2017104307-appb-000004
下文以本发明的代表性化合物9为例说明本发明化合物的制备方法。
本发明合成路线1如下所示：
Figure PCTCN2017104307-appb-000005
根据路线1的方法，本发明合成了以下一些代表性的化合物：
表一：化合物结构式
Figure PCTCN2017104307-appb-000006
Figure PCTCN2017104307-appb-000007
根据本发明的又一方面，提供一种药物组合物，含有治疗有效量的式(I)化合物以及药学上可接受的载体和/或辅料，用于治疗各种癌症、感染、炎症和常规增殖性疾病，或治疗以出现迅速增殖性细胞为特征的其它疾病如银屑病和其它皮肤病。所述治疗有效量是指药用组合物所含通式(I)化合物的量足以产生临床期望的治疗效应，例如使用药者的肿瘤大小降低至临床可接受的范围内。
在一个具体实施方案中，本发明的化合物或药物组合物用于治疗肉瘤、癌和/或白血病。可将所述化合物或组合物单独或作为治疗方案的一部分应用的示例性病症包括纤维肉瘤、粘液肉瘤、脂肪肉瘤、软骨肉瘤、骨源性肉瘤、脊索瘤、血管肉瘤、内皮肉瘤、淋巴管肉瘤、淋巴管内皮肉瘤、滑膜瘤、间皮瘤、尤文氏瘤、平滑肌瘤、横纹肌肉瘤、结肠癌、胰腺癌、乳腺癌、卵巢癌、前列腺癌、鳞状细胞癌、基底细胞癌、汗腺癌、皮脂腺癌、乳突癌、乳头状腺癌、囊腺癌、髓样癌、支气管癌、肾细胞癌、肝细胞癌、胆管癌、绒毛膜癌、精原细胞瘤、胚胎性癌、威尔姆氏肿瘤、子宫颈癌、睾丸肿瘤、肺癌、小细胞肺癌、膀胱癌、上皮癌、神经胶质瘤、星细胞瘤、成神经管细胞瘤、颅咽管瘤、室鼓 膜瘤、松果体瘤、血管母细胞瘤、听觉神经瘤、少突神经胶质瘤、脑膜瘤、黑素瘤、成神经细胞瘤、以及视网膜母细胞癌症。
在某些具体实施方案中，本发明的化合物或组合物用于治疗诸如乳房、前列腺、肾、膀胱、或结肠组织形成的癌的病症。
在其他具体实施方案中，本发明的化合物或药物组合物用于治疗脂肪组织中发生的增殖性或瘤性疾病，如脂肪细胞肿瘤即脂肪瘤、纤维脂瘤、成脂母细胞瘤、脂肪过多症、棕色脂肪瘤(hibemomas)、血管瘤和/或脂肪肉瘤。
在其他具体实施方案中，也可以用所述组合物和化合物控制传染物和寄生物(如细菌、锥体虫、真菌等)。
本发明的药用组合物可以通过静脉内、皮内、肌内、皮下、口服或吸入等途径给药，其药用组合物的剂型可以是胃肠道用药制剂如片剂、胶囊剂、丸剂等，也可以是胃肠外用药制剂如注射剂、外用制剂等。
此外，本发明还提供治疗抗肿瘤和相关疾病的方法，该方法包括对患有癌症或相关病况的患者使用治疗有效量的所述通式(I)的化合物。
本发明也提供在患者中调节微管聚合的方法，该方法包括给药治疗有效量的至少一种本发明化合物或其药物学可接受的前药、盐、水合物、溶剂合物、晶体形式或非对映异构体。
本发明的化合物可以与化疗剂联合应用治疗肿瘤，实验表明本发明化合物与化疗剂联合使用时可增加肿瘤抑制效果，同时降低化疗剂的毒性。最优选的化合物为化合物(9)，化疗剂可以为通常治疗肿瘤所使用的常规化疗剂，在一个优选实施例中，将化合物(9)与多西他赛联合使用取得了非常好的治疗效果。
本发明的化合物可结合微管蛋白，可以抑制癌细胞的生长和增殖，对相应的白细胞与粒细胞几乎无影响，在对抗肿瘤增殖时，对免疫系统功能干扰较小，大大改进了抗肿瘤药物临床使用的弊端。这种化合物还可用于治疗其他与过度增殖相关的病症。
附图说明
图1为本发明的化合物的肿瘤增殖抑制作用；
图2为化合物(9)的抗肿瘤作用以及安全性实验结果，图中：
A：化合物对于裸小鼠抑制肿瘤增殖的抑制作用；B：化合物抑制裸小鼠肿瘤增殖时对动物体重的影响；C：化合物(9)以及与多西他赛联合连续给药后对于SD大鼠中性粒细胞计数的影响；
图3为本发明的化合物对于大鼠中性粒细胞计数的影响。
具体实施方式
本发明的化合物结构改造是在以天然产物Phenylahistin为先导化合物基础 上，并结合已报导的构效关系进行相应的结构改造。根据插烯原理，在哌嗪和苯环之间再插入一个双键，在电性分布上，犹如哌嗪双烯和苯环直接相连，再将不同的取代基引入到苯环上，可能会得到活性相似或者活性更强的化合物；而哌嗪环与吡唑之间再插入一个双烯键使化合物更稳定，不容易在常温下变质，因此本发明化合物的稳定性更好，安全性更高。
下文以具体实施例进一步说明本发明，但不对本发明的范围形成任何限制。
通用方法：在RT-1熔点仪(天津分析仪器厂)设备上测量熔点，温度经校正。Bruker AV400(400MHz)光谱仪中记录1HNMR光谱并且将其以TMS的百万分之一份(ppm)低磁场进行说明。在Nicolet Magna 550FT-IR付立叶分光光度计上记录红外光谱。用HP1100Esquire2000液相色谱/质谱联用仪记录质谱。在岛津UV2410紫外分光光度计记录UV光谱。在硅胶GF254高效板(烟台市芝罘硅胶开发试验厂)上进行TLC。WZZ-1S旋光测定仪(上海精密科学仪器有限公司)上进行旋光测定。
实施例按照路线1合成式(I)化合物
实施例1
步骤1：
Figure PCTCN2017104307-appb-000008
在5000ml干燥的三口烧瓶中加入镁屑(14.4g，0.60mol)和干燥的四氢呋喃(1L)，滴加1ml1，2-二溴乙烷引发反应。然后缓慢滴加溴乙醛缩乙二醇(100g，0.60mol)的四氢呋喃溶液(500ml)。滴加完毕后，搅拌两小时至镁屑消失。然后将5-叔丁基-1H-咪唑-4-甲醛(30g,0.20mol)的四氢呋喃溶液(500ml)缓慢滴加到上述溶液中，搅拌过夜。然后用浓盐酸调节至酸性(pH＝1)，并加热至60℃搅拌30分钟。旋蒸除去四氢呋喃，加入乙酸乙酯萃取。有机相合并用水洗两次，饱和食盐水洗一次，最后用无水硫酸钠干燥，浓缩。粗产物经柱层析纯化(石油醚/乙酸乙酯的体积比:2/1)得到黄色固体(17.8g,yield:50％)。ESI-MS:m/z＝179.2(M+H)；1H-NMR:(500MHz,CDCl3)δ9.62(d,1H),7.68(d,1H),7.59(s,1H),6.75(dd,1H),1.48(s,9H)。
步骤2：
Figure PCTCN2017104307-appb-000009
250ml干燥的三口烧瓶中加入DMF(100ml)，1,4-二乙酰基-哌嗪-2,5-二酮 (2.5g,17.6mmol)，3-(5-叔丁基-1H-咪唑-4-)丙烯醛(2.09g,11.7mmol)和碳酸铯(5.74g,17.6mmol)。混合物在氮气保护下，室温搅拌16小时。将反应液用乙酸乙酯稀释至1.5L，有机相用食盐水洗3次，无水硫酸钠干燥，过滤，浓缩。粗产物经柱层析纯化(洗脱剂DCM/MeOH的体积比:200/1至30/1)得到亮黄色固体(1.89g,yield:43％).ESI-MS:m/z＝317.1(M+H)；1H-NMR:(500MHz,DMSO-d6)δ11.83(s,1H),10.66(s,1H),8.46(s,1H),7.56(d,1H),7.32(t,1H),7.18(d,1H),6.84(d,1H),6.77(s,1H),4.28(s,2H),1.32(s,9H)。
步骤3：
Figure PCTCN2017104307-appb-000010
1L干燥的单口烧瓶中加入3-乙烯基苯甲醛(13.2g,0.10mol)，甲酰基亚甲基三苯基磷烷(33.5g,0.11mmol)和甲苯(200ml)。反应体系回流17小时然后浓缩。粗产物经柱层析纯化(洗脱剂：石油醚/乙酸乙酯的体积比:100/1至80/20)得到黄色固体(9.6g,yield:60％).ESI-MS:m/z＝159.2(M+H)。
步骤4：
Figure PCTCN2017104307-appb-000011
250ml干燥的单口烧瓶中加入DMF(100ml)，中间体B(1.58g,5mmol)，3-(3-乙烯基苯基)丙烯醛(1.59g,10mmol)和碳酸铯(3.26g,10mmol)。反应体系在80℃搅拌2小时，冷却至室温，然后到入冰水中。乙酸乙酯萃取，有机相用食盐水洗三次，无水硫酸钠干燥，过滤，浓缩。粗产物用石油醚/乙酸乙酯(5/1)洗涤，得到亮黄色固体(900mg,yield:43％).ESI-MS:m/z＝415.1(M+H)；1H-NMR:(500MHz,DMSO-d6)δ11.82(s,1H),10.81(bs,2H),7.78-7.70(m,3H),7.54-7.49(m,3H),7.40-7.36(m,2H),7.11-7.08(m,1H),6.89-6.86(m,1H),6.78-6.72(m,1H),6.66-6.64(m,1H),6.52-6.50(m,1H),6.90(d,J＝17Hz,1H),5.31(d,J＝11Hz,1H),1.34(s,9H).
实施例2～11
步骤1～2制备中间体B，同实施例1中步骤1～2。其他中间体及终产品制备如下表所示：
表二：其他化合物制备方法
Figure PCTCN2017104307-appb-000012
Figure PCTCN2017104307-appb-000013
Figure PCTCN2017104307-appb-000014
Figure PCTCN2017104307-appb-000015
Figure PCTCN2017104307-appb-000016
实施例12：
实验方法：应用HTRF kinEASE TK kit检测11个化合物对Lynα、AKT2、KDR、IKK-β、c-RAF酶的抑制活性，将待测化合物从100μM到0.1μM等10个浓度稀释，控制DMSO浓度为1％，每个浓度为复孔。
在Lynα，AKT2，KDR，IKK-β、c-RAF酶以10μM的浓度对11个化合物进行抑制活性筛查。结果如下表所示：
表三：11个化合物对5种激酶的IC50值
Figure PCTCN2017104307-appb-000017
结果表明这些化合物对于Lynα、AKT2、KDR、IKK-β、c-RAF具有不同程度的抑制作用。
实施例13：细胞水平抑制实验
细胞株来源：
HT-1080CAKi-1人纤维肉瘤细胞、AGS人胃腺癌细胞、A375人皮肤黑色素瘤细胞、Fadu人咽鳞癌细胞、NCI-H1975人肺腺癌细胞、SK-ES-1人骨肉瘤细胞、MDA-MB-231人乳腺癌细胞、MIAPaCa-2人胰腺癌细胞均来源于ATCC。
实验方法
第一天细胞铺板
1.使用胰酶将细胞从细胞培养盘上消化下，使用培养基重悬后测定细胞密度。
RPM1640培养基用于培养AGS细胞(INVITROGEN)
McCoys’5A培养基用于培养SK-ES-1细胞(INVITROGEN)
EMEM培养基用于培养HT-1080Caki-1和MDA-MB-231细胞(INVITROGEN)
胎牛清(GIBCO#10099-141)
胰蛋白酶(GIBCO#25200-072)
发光法细胞活力检测试验盒(promega#G7572)
96孔板(from Coming#3365)
96孔板黑色(Costar#3340)
底部白色贴膜(Per#6005199)
2.将调整密度后的细胞溶液以每孔90微升加入细胞实验板中。
3.将铺好的细胞培养板放入孵箱，在37摄氏度，5％的CO2的湿润条件下孵育24小时。
第二天加化合物
1.根据实验模板准备200倍浓度的参照化合物和待测试化合物溶液。
2.取7微升化合物溶液到133微升培养基中进行稀释。
3.取10微升稀释后的化合物溶液加入前一天准备好的细胞培养板中。
4.加入化合物的细胞培养板重新放回孵箱，在37摄氏度，5％的CO2的湿润条件下孵育72小时。
第五天结果检测
1.检测试剂：
发光法细胞活力检测试剂盒(promega #G7572)，将底物冻干粉和缓冲液混合，按30ul/孔的量加入96孔板中，在实验前30分钟放置于室温进行平衡。
2.细胞培养板每孔加入30微升检测试剂、摇板10分钟，诱导细胞裂解。
3.10分钟后将细胞培养板在室温下孵育2分钟以稳定发光信号。
4.使用Envision读板，读板时时间设定为0.5秒每孔。
在HT-1080、AGS、A375、Fadu、NCI-H1975、Caki-1、SK-ES-1、MDA-MB-231和MIA PaCa-2的9个细胞株上，用细胞增殖实验检测4个化合物对细胞的抑制水平。化合物从600nM起始，3倍稀释，复孔检测。结果如下表所示：
表四：4个化合物对细胞的抑制水平
Figure PCTCN2017104307-appb-000018
结果表明，上述化合物对于各考察肿瘤细胞株具有很强的抑制作用，并且与参考药物[(-)苯基阿夕斯丁](商业购买)对比，该实施例中化合物抑制肿瘤的强度远高于参考药物。结合实施例12中对不同酶的影响可以看出，本实施例中化合物发挥抑制肿瘤作用的浓度大幅度低于抑制酶的浓度，因此可以推断出本实施例中化合物在抑制肿瘤增殖时，除对本申请中提及到的酶有抑制作用外，对其他尚未考察的酶活力或细胞内分子尚有较强的抑制作用，可能会不同程度地影响多种细胞内外靶点发挥强效的协同抑制肿瘤作用。
实施例14：对肿瘤的增殖的抑制作用
实验方法
将MES-SA人子宫肉瘤细胞(来源ATCC)长期培养于低浓度的阿霉素中，保持充分的细胞活力，诱导细胞表达p-glycoprotein。培养10个月后，利用WB检测诱导表达结果。诱导成功后，利用无阿霉素培养基培养一周后，调整细胞浓度，将肿瘤细胞接种于裸鼠腋下，并待移植肿瘤增殖到一定体积后给予不同的药物进行治疗。利用空白小鼠分别给予不同药物不同剂量摸索出不影响动物体重增长的剂量即为给予移植肿瘤动物治疗的剂量(图1)。
结果表明，在给予不会降低动物体重的剂量下，不同的药物对于裸鼠移植肿瘤抑制率具有一定的差异。其中(-)苯基阿夕斯丁在无毒剂量下几乎单独使用无对抗肿瘤增殖的效果。而多西他赛(商业购买)在无明显毒性该剂量下药效作用较差。化合物9、化合物11、化合物15、化合物18在对动物体重增殖无明显影 响的剂量下能有效地发挥抗肿瘤增殖的作用。
肿瘤抑制率(％)＝(1-治疗组瘤重量/对照组瘤重量)*100％，根据图1所示，多西他赛的肿瘤抑制率接近80％，并且高于化合物9、化合物11、化合物15、化合物18。
本实施例中同时测试了在同等药效剂量下，药物对于不同组织器官血流分布的影响。给药后2小时，多西他赛组中包含肿瘤组织在内，血流的分布与空白溶媒组类似。而化合物9、化合物11、化合物15、化合物18以及(-)苯基阿夕斯丁对除肿瘤组织外的其他组织未发现有明显的干预组织血流分布的影响。对于肿瘤组织，化合物9、化合物11、化合物15、化合物18以及(-)苯基阿夕斯丁组与空白溶媒组比较，能显著降低肿瘤内血流分布。
实施例15：化合物的抗肿瘤作用以及安全性
对接种肿瘤细胞的裸小鼠分别给以空白溶媒、多西他赛、化合物9以及多西他赛与化合物9联合药物，于给药后不同的时间点测量肿瘤的体积并检测肿瘤体积变化过程，同时检测裸小鼠的体重变化，以评价药物的抗肿瘤效果以及毒性反应程度。另对正常SD大鼠，按照以上给药与分组方式，在给药前以及末次给药后检测血中性粒细胞计数水平，以评价药物对于中性粒细胞计数的影响程度。
实验结果表明，化合物9在体内能有效地抑制肿瘤的增殖，同时与多西他赛联合使用后能显著增加抗肿瘤活性，同时有效地降低了多西他赛单独应用时的整体不良反应，能显著降低多西他赛毒性引起的动物体重降低。在正常大鼠体内联合使用多西他赛与化合物9后，与单独使用多西他赛比较，中性粒细胞计数显著升高，有效地抑制了多西他赛引起的中性粒细胞计数降低的不良反应。综合比较，在不影响动物整体状态的剂量下，化合物9对抗肿瘤的效果显著优于多西他赛，即化合物9的肿瘤治疗窗显著地优于多西他赛，同时为进一步增加抗肿瘤效果，将多西他赛与化合物9联合应用能有效地降低多西他赛毒性反应，并增加抗肿瘤作用(图2)。
实施例16：对中性粒细胞计数的影响
实验方法
利用多剂量分别给予不同的药物对裸鼠移植肿瘤的抑制程度考察，摸索出各个药物的等效剂量，并通过物种间剂量折算后给予大鼠不同药物。即在给予大鼠相对同等药效学剂量下，考察药物对于大鼠中性粒细胞计数的影响。连续给予Wista大鼠2周后，对大鼠采血并称量各个给药组体重(图3)。
结果表明，在同等药效剂量下，连续使用各个化合物给予大鼠后，常规抗肿瘤药物多西他赛能显著降低大鼠体内中性粒细胞计数，同时实验中观察到动物整体状态不佳。(-)苯基阿夕斯丁与多西他赛相比，在同等药效学剂量下对中性粒细胞降低作用轻于多西他赛，但是中性粒细胞计数仍显著低于空白溶媒组与 化合物组。而化合物9、化合物11、化合物15、化合物18组在与其他药物同等药效剂量下，对中性粒细胞无显著影响。因此，化合物9、化合物11、化合物15、化合物18在有效对抗肿瘤增殖时，能有效避免对中性粒细胞降低的不良反应。动物体重的变化趋势与中性粒细胞计数变化百分率呈正相关。
实施例17:对微管功能的作用
在本研究中使用人脐静脉内皮细胞(来自Cambrex的HuVEC)，通过对α-微管素进行染色，对比秋水仙碱和紫杉醇评价了化合物9、11、15、18和叔丁基苯基阿夕斯丁对微管素的效果。
在所述HuVEC细胞中，暴露于化合物9、11、15、18、叔丁基苯基阿夕斯丁或秋水仙碱(均为2μM)30分钟，诱导了与DMSO对照中观测结果相对的微管解聚作用(表现为缺乏完整的微管结构)和细胞膜起疱(blebbing)(细胞凋亡的明显标志)，而紫杉醇在这些条件下不能诱导微管解聚作用。秋水仙碱是已知的微管解聚试剂而紫杉醇是微管素稳定剂。将CCD-27sk细胞暴露于化合9、11、15、18或秋水仙碱可观察到类似的结果。
实施例18：药物组合物
根据本发明的化合物可以与生理学可接受的载体或媒介结合以提供药物组合物，如形式为含有各种填料和粘结剂的片剂或胶囊的低压冻干粉末。相似地，化合物可以与其它药剂共同给药，共同给药应该表示至少对患者使用两种药剂以提供两种药剂结合的有益效果。例如，可以在一定时间内同时或按顺序使用药剂。可以广泛地由本领域技术人员选择经验确定组合物中化合物的有效剂量。另外，根据适应症和所需治疗效果，本发明的化合物可以单独使用或与一种或多种额外的药剂结合使用。由本发明设想的联合治疗包括，例如，本发明化合物和额外药剂在单一药物配方中的使用以及本发明化合物和额外药剂在分开药物配方中的使用。
本发明化合物可单独，或者与可药用载体或稀释剂，任选与已知辅药例如微晶纤维素一起在依据标准药物实践的药物组合物中施用给乳动物、优选人。化合物可口服给药或非胃肠道给药，包括静脉内、肌内、腹膜内、皮下、直肠和局部给药途径。
对于本发明化疗化合物的口服应用，所选的化合物可例如以片剂或胶囊的形式或者作为水溶液或悬浮液施用。对于口服片剂，通常加入常用载体包括乳糖和玉米淀粉，以及润滑剂例如硬脂酸镁。对于以胶囊形式口服施用，可使用的稀释剂包括乳糖和无水玉米淀粉，当需要口服使用的水悬浮液时，将活性组分与乳化剂和悬浮剂混合。如果需要的话，可加入一些甜味剂和/或矫味剂。对于肌内、腹膜内、皮下或静脉内应用，应当控制溶质的总浓度以使制剂等渗。
18.1含有化合物9的药物组合物
Figure PCTCN2017104307-appb-000019
18.2制剂的制备方法
按比例取本发明的活性化合物、崩解剂和填充剂过60～100目筛，混合均匀，用2～20％的聚维酮K30的乙醇溶液制软材，过20～50目筛制粒，40～90℃干燥，颗粒水分控制在3％以内，整粒后加入适量的润滑剂，混合均匀，压片，即得。
具体地，上述实施例的药物组合物也可通过如下方法制备：按50倍处方量称取化合物9、18、填充剂和崩解剂依次过60、80目筛，混合均匀，用2～20％聚维酮K30的50％乙醇溶液制软材，30目筛制粒，60℃干燥，颗粒水分控制在3％以内，20目筛整粒后加入适量的润滑剂，混合均匀，压片，即得产品。
实施例19稳定性考察
将获得的化合物9进行稳定性考察(10天的加速试验)，在40℃、60℃、湿度75％、92.5％、光照条件下对化合物的水分、纯度、最大单杂及总杂与0天的数据进行对比，结果显示获得的化合物稳定。而(-)苯基阿夕斯丁在60℃条件下，降解明显，说明高温对(-)苯基阿夕斯丁的稳定性有影响。
表五 化合物9影响因素试验结果
Figure PCTCN2017104307-appb-000020
表六 (-)苯基阿夕斯丁影响因素试验结果
Figure PCTCN2017104307-appb-000021
以上对本发明的详细描述并不限制本发明，本领域技术人员可以在此基础上做出各种改变和变形，只要不脱离本发明的精神，均应属于本发明权利要求定义的范围。

Claims (10)

  1. 如下通式(I)表示的化合物：
    Figure PCTCN2017104307-appb-100001
    其中：
    Figure PCTCN2017104307-appb-100002
    n独立表示0～5的整数，条件是n≤5，A表示单取代或多取代的基团，所述基团选自H、C1-C20烷基、C1-C20烃基、C1-C20酰氨基、C1-C20酰氧基、C1-C20烷酰基、C1-C20烷氧羰基、C1-C20烷氧基、C1-C20烷氨基、C1-C20烷羧氨基、芳酰基、芳烷酰基、羧基、氰基、卤素、羟基、硝基和甲基噻吩基。
  2. 如权利要求1所述的化合物，其特征在于所述通式(I)中n独立表示0～2的整数，条件是n≤2，A表示单取代或多取代的基团，所述基团选自H、乙烯基、甲基、三氟甲氧基、甲氧基、氰基、卤素和苯甲酰基。
  3. 如权利要求1或2所述的化合物，选自下述化合物：
    (3Z,6Z)-3-((E)-3-(5-叔丁基)-1H-咪唑基-4-基)丙烯亚基)-6-((E)-3-(3-乙烯基苯基)-2-丙烯亚基)哌嗪-2,5-二酮(9)
    Figure PCTCN2017104307-appb-100003
    (3Z,6Z)-3-((E)-3-(5-叔丁基)-1H-咪唑基-4-基)丙烯亚基)-6-((E)-3-(3-氟苯基)-2-丙烯亚基)哌嗪-2,5-二酮(10)
    Figure PCTCN2017104307-appb-100004
    (3Z,6Z)-3-((E)-3-(5-叔丁基)-1H-咪唑基-4-基)丙烯亚基)-6-((E)-3-苯基丙烯亚基)哌嗪-2,5-二酮(11)
    Figure PCTCN2017104307-appb-100005
    (3Z,6Z)-3-((E)-3-(5-叔丁基)-1H-咪唑基-4-基)丙烯亚基)-6-((E)-3-(2,3-二甲基苯基)-2-丙烯亚基)哌嗪-2,5-二酮(12)
    Figure PCTCN2017104307-appb-100006
    (3Z,6Z)-3-((E)-3-(5-叔丁基)-1H-咪唑基-4-基)丙烯亚基)-6-((E)-3-(3-三氟甲氧基苯基)-2-丙烯亚基)哌嗪-2,5-二酮(13)
    Figure PCTCN2017104307-appb-100007
    (3Z,6Z)-3-((E)-3-(5-叔丁基)-1H-咪唑基-4-基)丙烯亚基)-6-((E)-3-(2,5-二氟苯基)-2-丙烯亚基)哌嗪-2,5-二酮(14)
    Figure PCTCN2017104307-appb-100008
    (3Z,6Z)-3-((E)-3-(5-叔丁基)-1H-咪唑基-4-基)丙烯亚基)-6-((E)-3-(3-甲氧基苯基)-2-丙烯亚基)哌嗪-2,5-二酮(15)
    Figure PCTCN2017104307-appb-100009
    (3Z,6Z)-3-((E)-3-(5-叔丁基)-1H-咪唑基-4-基)丙烯亚基)-6-((E)-3-(3,5-二甲氧基苯基)-2-丙烯亚基)哌嗪-2,5-二酮(16)
    Figure PCTCN2017104307-appb-100010
    (3Z,6Z)-3-((E)-3-(5-叔丁基)-1H-咪唑基-4-基)丙烯亚基)-6-((E)-3-(3-氯苯 基)-2-丙烯亚基)哌嗪-2,5-二酮(17)
    Figure PCTCN2017104307-appb-100011
    (3Z,6Z)-3-((E)-3-(5-叔丁基)-1H-咪唑基-4-基)丙烯亚基)-6-((E)-3-(3-腈基苯基)-2-丙烯亚基)哌嗪-2,5-二酮(18)
    Figure PCTCN2017104307-appb-100012
    (3Z,6Z)-3-((E)-3-(5-叔丁基)-1H-咪唑基-4-基)丙烯亚基)-6-((E)-3-(3-苯甲酰基苯基)-2-丙烯亚基)哌嗪-2,5-二酮(19)
    Figure PCTCN2017104307-appb-100013
  4. 一种药物组合物，包括治疗有效量的权利要求1至3任一项所述的化合物。
  5. 如权利要求4所述的药物组合物，其特征在于进一步含有药学上可接受的载体和/或辅料。
  6. 一种用于制备权利要求1所述的化合物的中间体，具有式(B)结构：
    Figure PCTCN2017104307-appb-100014
  7. 权利要求1所述的化合物在制备治疗肿瘤药物中的应用。
  8. 权利要求1所述的化合物作为微管蛋白抑制剂的用途。
  9. 权利要求1所述的化合物与化疗剂联合在制备治疗肿瘤药物中的应用。
  10. 权利要求9所述的应用，其中所述化合物为(3Z,6Z)-3-((E)-3-(5-叔丁基)-1H-咪唑基-4-基)丙烯亚基)-6-((E)-3-(3-乙烯基苯基)-2-丙烯亚基)哌嗪-2,5-二酮(9)，所述化疗剂为多西他赛。
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