WO2012163291A1 - Phenylpiperazine derivative for inhibiting tumor metastasis and tumor angiogenesis - Google Patents

Phenylpiperazine derivative for inhibiting tumor metastasis and tumor angiogenesis Download PDF

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WO2012163291A1
WO2012163291A1 PCT/CN2012/076384 CN2012076384W WO2012163291A1 WO 2012163291 A1 WO2012163291 A1 WO 2012163291A1 CN 2012076384 W CN2012076384 W CN 2012076384W WO 2012163291 A1 WO2012163291 A1 WO 2012163291A1
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pharmaceutically acceptable
tumor
group
cancer
acceptable salt
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PCT/CN2012/076384
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French (fr)
Chinese (zh)
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姜凤超
丁玉兰
蒋杰
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广州翰鼎医药科技有限公司
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D295/00—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms
    • C07D295/16—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms acylated on ring nitrogen atoms
    • C07D295/18—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms acylated on ring nitrogen atoms by radicals derived from carboxylic acids, or sulfur or nitrogen analogues thereof
    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D295/00—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms
    • C07D295/22—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms with hetero atoms directly attached to ring nitrogen atoms
    • C07D295/26—Sulfur atoms

Definitions

  • the invention belongs to the field of medicine, and particularly relates to a phenyl piperazine derivative which can further kill tumor by inhibiting tumor angiogenesis, thereby inhibiting tumor metastasis and inhibiting tumor growth, and a preparation method of the derivative and the intermediate thereof, It also relates to the use of such derivatives in the preparation of tumor suppressor angiogenesis, prevention of tumor metastasis, and inhibition of tumor growth, and as an adjunct to general tumor chemotherapy drugs and surgical treatment. Background technique
  • the tumor may have metastasized and spread due to the invasion of the tumor cells, resulting in a poor therapeutic effect, which ultimately leads to treatment failure. Therefore, controlling the spread and metastasis of tumor cells, whether during surgery or chemotherapy, is considered to be the key to complete treatment of tumors.
  • Angiogenesis plays an extremely important role in the multistep process of tumor invasion and metastasis.
  • Tumor angiogenesis is a complex process involving multiple cell proliferation, migration, apoptosis, integration, degradation, and structural remodeling. Targeting angiogenesis and biochemical changes during its development, angiogenesis inhibitors are developed. Controlling tumor growth and metastasis may become an important way to prevent and treat cancer.
  • Anti-angiogenesis therapy is a novel anti-tumor therapy that controls tumor growth by inhibiting angiogenesis. It can achieve early intervention, block or break the growth of new blood vessels to cut off the blood supply of the tumor, starvation of tumors to suppress tumor cells. In particular, the purpose of tumor cell proliferation after metastasis and diffusion.
  • integrins mainly mediate the mutual adhesion of cells and cells, cells and extracellular matrix (ECM), two-way signal transduction, cell adhesion, proliferation Differentiation, metastasis, and apoptosis play an important regulatory role.
  • ECM extracellular matrix
  • the integrin ⁇ 3 receptor is highly specific in the neovascular endothelial cell membrane in tumor tissues, and when its function is inhibited, vascular endothelial cell apoptosis, tumor growth and angiogenesis are inhibited.
  • integrin ⁇ 3 Receptor antagonists can significantly prevent tumor metastasis and recurrence, and even cause tumor regression. Summary of the invention
  • one of the objects of the present invention is to provide a class of phenylpiperazine derivatives and pharmaceutically acceptable salts thereof, which are integrin ⁇ 3 receptor antagonists, Higher inhibitory activity.
  • Another object of the present invention is to provide a use of the above phenylpiperazine derivatives for the preparation of a medicament for inhibiting tumor angiogenesis, preventing tumor metastasis, and inhibiting tumor growth, and as an auxiliary drug for general tumor chemotherapy drugs and surgical treatment.
  • a phenylpiperazine derivative and a pharmaceutically acceptable salt thereof which has a structure as shown in Formula I:
  • the R is a C1 to C11 chain hydrocarbon group or a cyclic hydrocarbon group having a 3 to 7 membered ring; the halogen is -F, -C1 or
  • R 3 is -H, -R, -OH, -OR, -COOH, -COOR, halogen or -CN group, wherein R is a fluorenyl group; n is a natural number between 0 and 5, more preferably 1.
  • the pharmaceutically acceptable salt is a sodium salt or an ammonium salt.
  • a pharmaceutical composition comprising the phenylpiperazine derivative of claim 1 and a pharmaceutically acceptable Acceptable carrier.
  • the compound of the present invention also has a certain effect on diseases associated with the integrin ⁇ 3 receptor.
  • the compound of the present invention has a suitable molecular weight, a stable structure, can penetrate a cell membrane, and has less toxic side effects.
  • phenylpiperazine derivatives and pharmaceutically acceptable salts thereof are useful for the preparation of adjunctive drugs for use as tumor chemotherapy drugs and surgical treatments.
  • the medicament is formulated into tablets, capsules, granules, fine granules, powders, pills, patches, oral liquids or injections.
  • the synthesis route of the above phenylpiperazine derivatives and intermediates is not complicated, the raw materials are easily available, are easy to be separated and purified, and the production cost is low.
  • the synthesis of the phenylpiperazine derivatives is to synthesize the related intermediate 1 and intermediate 2, such as the hydroxyhalogenation of diethanolamine, and the obtained bis(2-dichloroethyl)amine hydrochloride and substituted aromatic amine Cyclization under microwave or conventional conditions, storage of substituted aromatic piperazine (intermediate 1); aldehyde and ethyl acetoacetate are condensed in the presence of a base catalyst, the obtained product is saponified in the presence of a strong base, and decarboxylated to obtain a glutaric acid derivative.
  • the related intermediate 1 and intermediate 2 such as the hydroxyhalogenation of diethanolamine
  • the obtained bis(2-dichloroethyl)amine hydrochloride and substituted aromatic amine Cyclization under microwave or
  • the preparation of intermediates includes the preparation of substituted phenylpiperazines and the preparation of substituted phenylcycloglutaric anhydride.
  • the substituted phenylpiperazine can be synthesized by a microwave method and a conventional method, and the reaction formula is represented by the formula (1), wherein -H (hydrogen), -R, -OR, -COOR (carboxylate group), halogen or - A CN group, wherein R represents a thiol group, and the position of substitution on the phenyl ring is in the para, ortho or meta position of the piperazinyl group, respectively.
  • Microwave catalysis bis(2-chloroethyl)amine hydrochloride and substituted aniline solid (molar ratio 1: 1.2) Add to n-butanol and mix well. Microwave irradiation at 195w for 2 ⁇ 8 min. After cooling, add the calculated amount of powdered anhydrous sodium carbonate, continue microwave irradiation for 15 ⁇ 20 min, filter while hot, and settle the filtrate. Crystalline, substituted phenyl piperazine.
  • R1 is a fluorenyl group and a decyloxy group: the reaction formula is as shown in (3)
  • R1 is a carboxylate group: The reaction formula is as shown in (3)
  • R2 is a sulfonate group: The reaction formula is as shown in (4)
  • the present invention has the following advantages and beneficial effects:
  • the novel phenylpiperazine derivative I of the present invention is fully confirmed in the related experiments for its inhibition and killing effect on tumor cell growth such as liver cancer and non-small cell lung cancer. Since the goal of the present invention is closely related to angiogenesis closely related to tumor cell production and proliferation, Over-inhibition of this receptor not only inhibits the growth and development of tumor cells, but also directly and indirectly kills tumor cells. It also plays a role in the spread and metastasis of tumor cells, and can also be used as an existing anti-tumor drug. Auxiliary drugs produce synergistic effects and produce stronger effects.
  • the novel structure of the phenylpiperazine derivative I mentioned in the present invention is advantageous for the exertion of the pharmacological activity of the derivative and the improvement of the pharmacokinetic properties.
  • the synthesis route is simple, the raw materials are easy to obtain, the post-treatment is simple, the purification is easy, and the cost is also one of the advantages.
  • FIG. 1 is a graph showing the inhibitory activity of phenylpiperazine derivatives JA3031 (1), JA3024 (2), JA3040 (3 ) and JA3008 (4) on human umbilical vein endothelial cells (ECV304) according to an embodiment of the present invention. A histogram of the test results.
  • FIG. 2 is a diagram showing the phenylpiperazine derivative compounds JA3031 (1), JA3024 (2), JA3040 (3 ), JA3036 (4), JA3023 (5 ) and JA3018 (6) provided according to an embodiment of the present invention.
  • FIG. 3 is a diagram showing the phenylpiperazine derivative compounds JA3031 (1), JA3024 (2), JA3008 (7), JA3031 (8), JA3104 (9) for hepatoma cells HepG-2 according to an embodiment of the present invention.
  • Fig. 4 is a bar graph depicting the results of an experiment for inhibiting the growth of mouse S-180 tumors by the phenylpiperazine derivatives JA303 JA3024, JA3036 alone or in combination according to an embodiment of the present invention.
  • Example 8 Synthesis and implementation of 3-(3,5-dimethoxyphenyl)-5-(4-(4-ethoxycarbonyl)phenyl)piperazine)-5-oxo-3-pentanoic acid (JA3023) Similarly to Example 7, the reactant was 1-(4-ethoxyformylphenyl)piperazine hydrochloride, 3-(3,5-dimethoxyphenyl)cycloglutaric anhydride.
  • a pharmacological model of human umbilical vein endothelial cell (ECV304) proliferation comparable to tumor vascular endothelial cells was selected for preliminary screening of the target product. MTT reduction method was used to detect cell proliferation and apoptosis.
  • Sample solution configuration According to the pre-experiment of the compound, the activity inhibition rate of the sample of 22.5 mol/L is about 60%.
  • the experimental design concentrates the concentration of the solution in the desired compound with a desired inhibition rate of 50%, that is, 30 ⁇ 1, and the other lower concentration.
  • the stock solution of 200 ⁇ /L was diluted with 1:4, 1:8, 1:40, 1:200, 1:2000 to obtain the desired five concentration gradients: 5 ( ⁇ mol/L) 25 mol/L, 5 mol/L, ⁇ /L, 0.1 mol/L.
  • Control design 5 ( ⁇ mol/L 5-Fu (50 ⁇ 1/ ⁇ ) as a positive control, 1.25% DMSO as a blank control, in order to avoid interference caused by MTT and environmental factors, set 8 complexes for each concentration gradient, try to Reduce the error.
  • the ECV304 cryotube was taken out from the liquid nitrogen tank and quickly placed in warm water at 37 ° C. After 5 minutes, the cryotube was placed in a centrifuge tube and centrifuged (1000 r/min) for 5 minutes. , suck the upper layer For the liquid, add an appropriate amount of serum-free RPMI-1640 medium, blow the cells with a pipette, centrifuge (1000r/min) for 5 minutes, and aspirate the supernatant.
  • RPMI-1640 medium containing 10% fetal calf serum, 100 U/ml penicillin and 10 ( ⁇ g/ml streptomycin) was added to the cells, and the cells were blown off with a pipette and inoculated into a 50 ml culture flask. 37 ° C, 5
  • the culture was carried out in an incubator of % 0 2 and changed every other day. On the fourth day, cell fusion was carried out, and the shape was cobblestone shape, subcultured.
  • Seed plate The cells were separately diluted with trypsin, containing 10% fetal bovine serum RPMI-1640 medium, and inoculated into 96-well culture plates at 37 ° C, 5 ⁇ M per well (10 4 cells). Incubate in the %C0 2 incubator until the cells are attached.
  • Loading Add the sample to be tested (100 ⁇ 7 wells). Incubate in a 37 ° C incubator for 72 h.
  • Color development Add 20 ⁇ l of sputum solution (5 mg/mL PBS solution per well, filter and sterilize, store at 4 °C in the dark), and continue incubation for 4 hours in the incubator.
  • Test Drain the cell culture medium, add 10 ( ⁇ L DMSO per well, shake for 10 min, and wait until the formazan is fully dissolved, and measure the OD value by the microplate reader.
  • Sample solution configuration 5mg of compound was weighed separately, and sterilized by UV method for 20min, dissolved in 0.4ml of DMSO (Sigma), added dropwise to 9ml of RPMI-1640 cell culture medium, sonicated, and then added.
  • the total volume of 0.6 ml of RPMI-1640 cell culture medium was 10 ml.
  • the HepG-2 cryotube was taken out from the low temperature freezer and placed in a 37 ° C water bath. After 10 minutes, it was taken into the cell room and placed on a clean bench to transfer the culture solution in the cryotube.
  • centrifuge 1000r / min
  • the prepared medium containing 50ml of the medium
  • blow the cells with a pipette and then inoculate 50ml of culture.
  • the flask was placed in a carbon dioxide incubator (37 ° C, 5% CO 2 ) for culture, and the cells were adhered to the next day, and the medium was changed to continue the culture.
  • Seed plate Place the culture bottle on the clean bench, pour the culture medium, remove 1 ml of trypsin into the culture flask, lay it flat for 30 s, then pour the trypsin, and then place the culture flask into the carbon dioxide incubator. Incubate in the medium. After 3 minutes, the culture flask is placed under the microscope to observe that the cells have been digested and floated. Add 5 ml of the medium to the flask, transfer 2 ml into a 50 ml centrifuge tube, add the medium and dilute to 25 ml. For the plate; 2 ml of the medium was added to the flask, and the culture was carried out in the incubator for a second measurement.
  • Adding 6 target compounds and 5-Fu to add 6 concentrations add the drug stock solution to 100 ⁇ 1, 50 ⁇ 25 ⁇ 1, 15 ⁇ 1, ⁇ and 5 ⁇ 1, respectively, and then add the corresponding amount of RPMI-1640 cell culture solution per well to make each well. Dilute to ⁇ ⁇ , the concentration of the sample obtained by dilution is 0.5mg/ml, 0.25mg/ml, 0.125 mg/mK 0.075 mg/ml, 0.05 mg/ml and 0.025 mg/ml, and each concentration is set to 5 complexes. Empty; set 10 holes as a blank control. Incubate in a 37 ° C incubator for 48 h.
  • DMSO dimethyl sulfoxide
  • Inhibition-control group average tumor weight group average tumor weight 100
  • the average tumor weight of the control group is shown in Table 1.
  • Table 1 Inhibition of tumor growth by mouse S-180 by some compounds and combination

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Abstract

Disclosed are a phenylpiperazine derivative and pharmaceutically acceptable salts thereof, characterized in that the derivative has the structure as represented in formula (I); R1 in the formula is a -H, -R, -OR, -COOR, halogen, or -CN group, and R represents alkyl; the substitution of R1 on the benzene ring is at the para position, ortho position, or meta position of the piperazinyl; m is a natural number from 0 to 2; R2 is a substituted or unsubstituted carboxylic acid group or sulfonic acid group. The phenylpiperazine derivative and pharmaceutically acceptable salts thereof can be used in preparing medications for inhibiting tumor angiogenesis and tumor metastasis, in preparing tumor medications for treating liver cancer, breast cancer, ovarian cancer, stomach cancer, colon cancer, lung cancer, or melanoma, and in preparing auxiliary medications for tumor chemotherapy and surgery treatments.

Description

用于抑制肿瘤转移和肿瘤血管生长的苯基哌嗪类衍生物 技术领域  Phenylpiperazine derivatives for inhibiting tumor metastasis and tumor blood vessel growth
本发明属于医药领域， 尤其涉及一种通过抑制肿瘤血管生成、 从而抑制肿 瘤转移并抑制肿瘤生长进一步杀灭肿瘤的苯基哌嗪类衍生物， 该类衍生物及其 中间体的制备方法， 同时还涉及该类衍生物在制备抑制肿瘤血管生成、 预防肿 瘤转移及抑制肿瘤生长、 以及在作为一般肿瘤化疗药物和手术治疗中的辅助药 物中的用途。 背景技术  The invention belongs to the field of medicine, and particularly relates to a phenyl piperazine derivative which can further kill tumor by inhibiting tumor angiogenesis, thereby inhibiting tumor metastasis and inhibiting tumor growth, and a preparation method of the derivative and the intermediate thereof, It also relates to the use of such derivatives in the preparation of tumor suppressor angiogenesis, prevention of tumor metastasis, and inhibition of tumor growth, and as an adjunct to general tumor chemotherapy drugs and surgical treatment. Background technique
往往在患者被检查出肿瘤时， 由于肿瘤细胞的侵润作用， 肿瘤可能已经发 生转移与扩散， 导致治疗效果欠佳， 最终导致治疗失败。 因此， 无论在对肿瘤 进行手术治疗还是化疗过程中， 控制肿瘤细胞的扩散和转移都被认为是完整治 疗肿瘤的关键。  Often, when a patient is examined for a tumor, the tumor may have metastasized and spread due to the invasion of the tumor cells, resulting in a poor therapeutic effect, which ultimately leads to treatment failure. Therefore, controlling the spread and metastasis of tumor cells, whether during surgery or chemotherapy, is considered to be the key to complete treatment of tumors.
由于类型的差别， 导致肿瘤转移和扩散的机制和程度不同， 因此迄今为止， 真正用于临床的具有较强的抑制肿瘤转移作用的药物还很少见。 在肿瘤发生侵 袭转移的多步骤过程中， 血管新生 （angiogenesis) 扮演着极为重要的角色。 肿 瘤血管生成是一个涉及多种细胞增殖、 迁徙、 凋亡、 整合、 降解及结构重塑的 复杂过程， 以血管生成的各个环节及其发生过程中的生化改变为靶点， 研制血 管生成抑制剂， 控制肿瘤生长和转移， 有可能成为肿瘤防治的一个重要途径。 抗血管新生疗法是通过抑制血管新生来控制肿瘤生长的新型抗肿瘤疗法， 它能 达到早期干预、 阻断或破环新生血管的生长以切断肿瘤的血液供应、 饿死肿瘤 的方式达到遏制肿瘤细胞尤其是转移和扩散后的肿瘤细胞增殖的目的。  Due to the difference in types, the mechanisms and extent of tumor metastasis and spread are different. Therefore, drugs that have been clinically used to inhibit tumor metastasis have been rare. Angiogenesis plays an extremely important role in the multistep process of tumor invasion and metastasis. Tumor angiogenesis is a complex process involving multiple cell proliferation, migration, apoptosis, integration, degradation, and structural remodeling. Targeting angiogenesis and biochemical changes during its development, angiogenesis inhibitors are developed. Controlling tumor growth and metastasis may become an important way to prevent and treat cancer. Anti-angiogenesis therapy is a novel anti-tumor therapy that controls tumor growth by inhibiting angiogenesis. It can achieve early intervention, block or break the growth of new blood vessels to cut off the blood supply of the tumor, starvation of tumors to suppress tumor cells. In particular, the purpose of tumor cell proliferation after metastasis and diffusion.
作为细胞黏附分子家族的重要成员之一的整合素， 主要介导细胞与细胞、 细胞与细胞外基质 （extracellular matrix, ECM) 之间的相互粘附的双向信号传 导， 对细胞的粘附、 增殖、 分化、 转移、 凋亡起重要调控作用。 整合素 ανβ3受 体在肿瘤组织中的新生血管内皮细胞膜有高度特异性表达， 当其功能受到抑制 时，血管内皮细胞凋亡，肿瘤生长及血管生成受到抑制。实验证明，整合素 ανβ3 受体拮抗剂能明显预防肿瘤转移及复发， 甚至使肿瘤消退。 发明内容 As an important member of the family of cell adhesion molecules, integrins mainly mediate the mutual adhesion of cells and cells, cells and extracellular matrix (ECM), two-way signal transduction, cell adhesion, proliferation Differentiation, metastasis, and apoptosis play an important regulatory role. The integrin ανβ3 receptor is highly specific in the neovascular endothelial cell membrane in tumor tissues, and when its function is inhibited, vascular endothelial cell apoptosis, tumor growth and angiogenesis are inhibited. Experiments show that integrin ανβ3 Receptor antagonists can significantly prevent tumor metastasis and recurrence, and even cause tumor regression. Summary of the invention
为了解决上述现有技术中存在的不足之处， 本发明的目的之一在于提供一 类苯基哌嗪类衍生物及其药学上可接受的盐， 其为整合素 ανβ3受体拮抗剂， 具 有较高的抑制活性。  In order to solve the above-mentioned deficiencies in the prior art, one of the objects of the present invention is to provide a class of phenylpiperazine derivatives and pharmaceutically acceptable salts thereof, which are integrin ανβ3 receptor antagonists, Higher inhibitory activity.
本发明的另一目的在于提供上述苯基哌嗪类衍生物在制备抑制肿瘤血管生 成、 预防肿瘤转移及抑制肿瘤生长、 以及在作为一般肿瘤化疗药物和手术治疗 中的辅助药物中的用途。  Another object of the present invention is to provide a use of the above phenylpiperazine derivatives for the preparation of a medicament for inhibiting tumor angiogenesis, preventing tumor metastasis, and inhibiting tumor growth, and as an auxiliary drug for general tumor chemotherapy drugs and surgical treatment.
本发明的目的通过下述技术方案实现： 一种苯基哌嗪类衍生物及其药学上 可接受的盐， 该衍生物具有如式 I所示的结构：
Figure imgf000004_0001
The object of the present invention is achieved by the following technical means: A phenylpiperazine derivative and a pharmaceutically acceptable salt thereof, which has a structure as shown in Formula I:
Figure imgf000004_0001
I  I
式中 (氢）、 -R、 -OR, -COOR (羧酸酯基）、 卤素或 -CN基团， 其 中 R代表垸基， 在苯环上的取代位置分别在哌嗪基的对位、 邻位或间位； m 为 0〜2之间的自然数， 更加优选为 0; R2为取代或未取代的羧酸基团或磺酸基 团。 Wherein (hydrogen), -R, -OR, -COOR (carboxylate group), halogen or -CN group, wherein R represents a fluorenyl group, the substitution position on the phenyl ring is in the para position of the piperazinyl group, Ortho or meta; m is a natural number between 0 and 2 , more preferably 0; R 2 is a substituted or unsubstituted carboxylic acid group or a sulfonic acid group.
所述 R为 C1〜C11链烃基或带有 3〜7元环的环烃基； 所述卤素为 -F、 -C1或 The R is a C1 to C11 chain hydrocarbon group or a cyclic hydrocarbon group having a 3 to 7 membered ring; the halogen is -F, -C1 or
-Br。
Figure imgf000004_0002
-Br.
Figure imgf000004_0002
R3为 -H、 -R、 -OH、 -OR、 -COOH、 -COOR、 卤素或 -CN基团， 其中 R为 垸基； n为 0〜5之间的自然数， 更加优选为 1。 R 3 is -H, -R, -OH, -OR, -COOH, -COOR, halogen or -CN group, wherein R is a fluorenyl group; n is a natural number between 0 and 5, more preferably 1.
所述药学上可接受的盐为钠盐或铵盐。  The pharmaceutically acceptable salt is a sodium salt or an ammonium salt.
一种药物组合物， 其包含了权利要求 1所述的苯基哌嗪类衍生物和药学上 可接受的载体。 A pharmaceutical composition comprising the phenylpiperazine derivative of claim 1 and a pharmaceutically acceptable Acceptable carrier.
另外， 本发明的苯基哌嗪类化合物是以整合素 ανβ3受体为靶标， 因此本发 明的化合物对于与整合素 ανβ3受体有关的疾病也具有一定的作用。  Further, since the phenylpiperazine compound of the present invention targets the integrin ανβ3 receptor, the compound of the present invention also has a certain effect on diseases associated with the integrin ανβ3 receptor.
本发明所涉及的化合物分子量合适， 结构稳定， 可穿透细胞膜， 毒副作用 较小。  The compound of the present invention has a suitable molecular weight, a stable structure, can penetrate a cell membrane, and has less toxic side effects.
上述苯基哌嗪类衍生物及其药学上可接受的盐在制备抑制肿瘤血管生成和 肿瘤细胞转移方面药物中的用途。  Use of the above phenylpiperazine derivatives and pharmaceutically acceptable salts thereof for the preparation of a medicament for inhibiting tumor angiogenesis and tumor cell metastasis.
上述苯基哌嗪类衍生物及其药学上可接受的盐在制备治疗肝癌、 乳腺癌、 卵巢癌、 胃癌、 结肠癌、 肺癌或黑色素瘤肿瘤药物中的用途。  The use of the above phenylpiperazine derivative and a pharmaceutically acceptable salt thereof for the preparation of a medicament for treating liver cancer, breast cancer, ovarian cancer, gastric cancer, colon cancer, lung cancer or melanoma tumor.
上述苯基哌嗪类衍生物及其药学上可接受的盐在制备作为肿瘤化疗药物和 手术治疗中的辅助药物中的用途。  The above phenylpiperazine derivatives and pharmaceutically acceptable salts thereof are useful for the preparation of adjunctive drugs for use as tumor chemotherapy drugs and surgical treatments.
所述药物制成片剂、 胶囊剂、 颗粒剂、 细粒剂、 粉剂、 丸剂、 贴剂、 口服 液或注射剂。  The medicament is formulated into tablets, capsules, granules, fine granules, powders, pills, patches, oral liquids or injections.
上述苯基哌嗪类衍生物以及中间体的合成路线不复杂， 原料易得， 易于分 离和纯化， 生产成本较低。 苯基哌嗪类衍生物的合成是先合成相关中间体 1 和 中间体 2， 如二乙醇胺的羟基卤代后， 得到的双 (2-二氯乙基)胺盐酸盐与取代芳 香胺在微波或常规条件下环化， 贮备出取代芳香哌嗪 （中间体 1 ); 醛和乙酰乙 酸乙酯在碱催化剂存在下缩合， 所得产物在强碱存在下皂化， 脱羧得戊二酸衍 生物， 后者在酸酐存在下脱水形成酸酐 （中间体 2); 中间体 1和中间体 2或者 其他的已知酸酐反应， 得目标产物 I， 即本发明苯基哌嗪类衍生物。 具体包括以 下步骤：  The synthesis route of the above phenylpiperazine derivatives and intermediates is not complicated, the raw materials are easily available, are easy to be separated and purified, and the production cost is low. The synthesis of the phenylpiperazine derivatives is to synthesize the related intermediate 1 and intermediate 2, such as the hydroxyhalogenation of diethanolamine, and the obtained bis(2-dichloroethyl)amine hydrochloride and substituted aromatic amine Cyclization under microwave or conventional conditions, storage of substituted aromatic piperazine (intermediate 1); aldehyde and ethyl acetoacetate are condensed in the presence of a base catalyst, the obtained product is saponified in the presence of a strong base, and decarboxylated to obtain a glutaric acid derivative. The latter is dehydrated in the presence of an acid anhydride to form an acid anhydride (Intermediate 2); Intermediate 1 and Intermediate 2 or other known anhydrides are reacted to give the desired product I, i.e., the phenylpiperazine derivative of the present invention. Specifically, the following steps are included:
Α、 中间体制备  Preparation of intermediates
其中中间体的制备包括了取代苯基哌嗪的制备和取代苯基环戊二酸酐的制 备。  The preparation of intermediates includes the preparation of substituted phenylpiperazines and the preparation of substituted phenylcycloglutaric anhydride.
(a) 取代苯基哌嗪的合成  (a) Synthesis of substituted phenylpiperazine
取代苯基哌嗪可利用微波法和常规法合成， 反应式如式（1 )所示， 其中 为 -H (氢）、 -R、 -OR, -COOR (羧酸酯基）、 卤素或 -CN基团， 其中 R代表垸 基， 在苯环上的取代位置分别在哌嗪基的对位、 邻位或间位。  The substituted phenylpiperazine can be synthesized by a microwave method and a conventional method, and the reaction formula is represented by the formula (1), wherein -H (hydrogen), -R, -OR, -COOR (carboxylate group), halogen or - A CN group, wherein R represents a thiol group, and the position of substitution on the phenyl ring is in the para, ortho or meta position of the piperazinyl group, respectively.
( 1 )微波催化法：将双 (2-氯乙基)胺盐酸盐与取代苯胺固体（摩尔比 1 : 1.2) 加入到正丁醇中搅拌混合均匀， 195w功率下微波辐射 2〜8 min， 稍冷却后加入 计算量的粉状无水碳酸钠，继续微波辐射 15〜20 min，趁热过滤，滤液静置析晶， 得取代苯基哌嗪。 (1) Microwave catalysis: bis(2-chloroethyl)amine hydrochloride and substituted aniline solid (molar ratio 1: 1.2) Add to n-butanol and mix well. Microwave irradiation at 195w for 2~8 min. After cooling, add the calculated amount of powdered anhydrous sodium carbonate, continue microwave irradiation for 15~20 min, filter while hot, and settle the filtrate. Crystalline, substituted phenyl piperazine.
(2) 常规法： 将双 (2-氯乙基)胺盐酸盐加入到正丁醇搅拌混合均匀， 搅拌 下加入取代苯胺固体（与双 (2-二氯乙基)胺盐酸盐的摩尔比 1 : 1.2)， 加热回流搅 拌反应 30〜60h， 稍冷却后加入计算量的粉状无水碳酸钠， 继续回流搅拌反应 48〜150h， 用 TLC监测反应至反应完全，， 趁热过滤， 滤液静置析晶， 得取代苯  (2) Conventional method: Add bis(2-chloroethyl)amine hydrochloride to n-butanol, mix well, and add substituted aniline solid (with bis(2-dichloroethyl)amine hydrochloride) with stirring. Molar ratio 1: 1.2), stirred under reflux with heating for 30~60h. After cooling, add the calculated amount of powdered anhydrous sodium carbonate, continue to reflux and stir the reaction for 48~150h. The reaction is monitored by TLC until the reaction is complete. The filtrate is allowed to stand for crystallization, and the substituted benzene is obtained.
Figure imgf000006_0001
Figure imgf000006_0001
( 1 )  ( 1 )
(b) 取代苯基环戊二酸酐的合成 ： 反应式如式 （1 ) 所示， 其中 为-11、 -R、 -OH、 -OR、 -COOH、 -COOR、 卤素或 -CN基团， 其中 R为垸基。 (b) Synthesis of substituted phenylcycloglutaric anhydride: The reaction formula is represented by formula (1) wherein -11, -R, -OH, -OR, -COOH, -COOR, halogen or -CN group, Wherein R is a thiol group.
( 1 ) 取代苯基戊二酸二乙酯的制备： 将取代苯甲醛和乙酰乙酸乙酯 （摩 尔比 1:2)， 乙醇混合均匀， 室温搅拌下向上述混合液中缓慢滴加催化剂量的六 氢吡啶溶液， 20〜50°C水浴下反应 1 h后析出大量固体，继续搅拌 2 h，减压过滤， 干燥得取代苯基戊二酸二乙酯；  (1) Preparation of substituted phenyl glutaric acid diethyl ester: The substituted benzaldehyde and ethyl acetoacetate (molar ratio 1:2), ethanol are uniformly mixed, and the amount of catalyst is slowly added dropwise to the above mixture under stirring at room temperature. The hexahydropyridine solution was reacted in a water bath at 20 to 50 ° C for 1 h, and a large amount of solid was precipitated. Stirring was continued for 2 h, filtered under reduced pressure, and dried to give diethyl phenyl glutarate.
(2)取代苯基戊二酸的制备： 将取代苯基戊二酸二乙酯（O.lmol)分批加 入到 100ml 50%KOH溶液中， 控制温度 30〜80°C下反应 2小时， 停止加热， 冷 却至室温， 控温下用酸将反应液调至 pH约等于 2-4， 过滤析出的固体， 用水洗 涤至滤液 pH=6， 干燥取代苯基戊二酸；  (2) Preparation of substituted phenyl glutaric acid: diethyl phenyl glutaric acid (0.1 mol) was added in portions to 100 ml of a 50% KOH solution, and the reaction was carried out at a temperature of 30 to 80 ° C for 2 hours. Stop heating, cool to room temperature, adjust the reaction solution to pH about 2-4 with acid under temperature control, filter the precipitated solid, wash with water until the filtrate pH=6, dry substituted phenylglutaric acid;
(3 ) 取代苯基环戊二酸酐的制备： 将取代苯基戊二酸 （O.lmol), 乙酸酐 (30ml) , 搅拌溶解， 加热回流反应 4〜10h， 冷却放置， 固体析出， 即为取代苯 基环戊二酸酐。 三步反应的总产率为 50〜70%。
Figure imgf000006_0002
(2) (3) Preparation of substituted phenylcycloglutaric anhydride: Substituted phenylglutaric acid (0.1 mol), acetic anhydride (30 ml), stirred and dissolved, heated to reflux for 4 to 10 h, placed under cooling, and solid precipitated. Substituted phenylcycloglutaric anhydride. The total yield of the three-step reaction is 50 to 70%.
Figure imgf000006_0002
(2)
B、 目标化合物 I苯基哌嗪类衍生物的制备 B. Preparation of target compound IPhenylpiperazine derivative
( 1 ) R1为垸基及垸氧基： 反应式如 （3 ) 所示  (1) R1 is a fluorenyl group and a decyloxy group: the reaction formula is as shown in (3)
将等摩尔的取代苯基哌嗪的盐酸盐和取代苯基环戊二酸酐， 催化量的吡啶， 用无水乙醇溶解， 在 130 w功率下微波辐射反应 20〜50分钟， 趁热过滤， 滤液 静置析晶。 过滤所得结晶， 无水乙醇重结晶， 产率在 50〜78%之间。  Equimolar substitution of phenylpiperazine hydrochloride and substituted phenylcycloglutaric anhydride, catalytic amount of pyridine, dissolved in absolute ethanol, microwave irradiation reaction at 130 w power for 20 to 50 minutes, hot filtered, The filtrate was allowed to stand for crystallization. The crystals obtained were filtered, and recrystallized from absolute ethanol in a yield of 50 to 78%.
(2) R1为羧酸酯基： 反应式如 （3 ) 所示  (2) R1 is a carboxylate group: The reaction formula is as shown in (3)
将等摩尔的取代苯基哌嗪和取代苯基环戊二酸酐， 再加入催化量的吡啶和 DMAP, 用丙酮溶解， 在 65 w功率下微波辐射， 趁热过滤， 滤饼用丙酮淋洗， 滤液静置析晶， 过滤， 所得固体用无水乙醇重结晶， 产率为 40〜70%。  Equimolar substitution of phenylpiperazine and substituted phenylcycloglutaric anhydride, addition of catalytic amount of pyridine and DMAP, dissolution with acetone, microwave irradiation at 65 w power, hot filtration, filter cake rinsed with acetone, The filtrate was allowed to stand for crystallisation and filtered, and the obtained solid was recrystallized from anhydrous ethanol to yield 40 to 70%.
(3 ) R2为磺酸酯基： 反应式如 （4) 所示  (3) R2 is a sulfonate group: The reaction formula is as shown in (4)
将 5 mmol取代苯基哌嗪溶于吡啶中， 室温搅拌下分批加入取代苯磺酰氯 6 mmol, 完全加入后控制外温 40°C加热过夜； 冷却， 过滤， 反应液滴入冰水中， 3h过滤， 所得固体用无水乙醇重结晶， 产率在 58〜70%之间。  5 mmol of substituted phenylpiperazine was dissolved in pyridine, and 6 mM of substituted benzenesulfonyl chloride was added in portions with stirring at room temperature. After heating, the external temperature was controlled at 40 ° C overnight; cooling, filtration, and the reaction was dropped into ice water for 3 h. After filtration, the obtained solid was recrystallized from absolute ethanol in a yield of 58 to 70%.
Figure imgf000007_0001
本发明相对于现有技术， 具有如下的优点及有益效果：
Figure imgf000007_0001
Compared with the prior art, the present invention has the following advantages and beneficial effects:
( 1 ) 本发明涉及的一类结构新颖的苯基哌嗪衍生物 I， 在相关的实验中充 分证实了其对于肝癌、 非小细胞肺癌等肿瘤细胞生长的抑制作用和杀灭作用。 由于本发明的目标是与肿瘤细胞生成和扩散密切相关的血管生成密切相关， 通 过抑制该受体， 不仅可以抑制肿瘤细胞的生长和发育， 具有直接和间接杀灭肿 瘤细胞的作用， 对于肿瘤细胞的扩散和转移也具有一定的作用， 同时也可以作 为现有抗肿瘤药物的辅助药物， 产生协同作用， 产生更强的作用。 (1) The novel phenylpiperazine derivative I of the present invention is fully confirmed in the related experiments for its inhibition and killing effect on tumor cell growth such as liver cancer and non-small cell lung cancer. Since the goal of the present invention is closely related to angiogenesis closely related to tumor cell production and proliferation, Over-inhibition of this receptor not only inhibits the growth and development of tumor cells, but also directly and indirectly kills tumor cells. It also plays a role in the spread and metastasis of tumor cells, and can also be used as an existing anti-tumor drug. Auxiliary drugs produce synergistic effects and produce stronger effects.
(2) 本发明所提到的结构新颖的苯基哌嗪衍生物 I， 有利于其衍生物药理 活性的发挥和药物动力学性质的改善。 另外， 与其他已知的整合素 ανβ3受体拮 抗剂等类型的抗肿瘤药物相比， 合成路线简单， 原料易得， 后处理简单， 易于 纯化， 成本低廉也是其优势之一。 附图说明  (2) The novel structure of the phenylpiperazine derivative I mentioned in the present invention is advantageous for the exertion of the pharmacological activity of the derivative and the improvement of the pharmacokinetic properties. In addition, compared with other known anti-tumor drugs such as integrin ανβ3 receptor antagonists, the synthesis route is simple, the raw materials are easy to obtain, the post-treatment is simple, the purification is easy, and the cost is also one of the advantages. DRAWINGS
图 1为描述根据本发明的实施方案所提供的苯基哌嗪衍生物 JA3031 ( 1 )、 JA3024 (2)、 JA3040 (3 ) 以及 JA3008 (4) 对于人脐静脉内皮细胞 （ECV304) 的抑制活性试验结果的柱状图。  1 is a graph showing the inhibitory activity of phenylpiperazine derivatives JA3031 (1), JA3024 (2), JA3040 (3 ) and JA3008 (4) on human umbilical vein endothelial cells (ECV304) according to an embodiment of the present invention. A histogram of the test results.
图 2为描述根据本发明的实施方案所提供的苯基哌嗪衍生物化合物 JA3031 ( 1 )、 JA3024 (2)、 JA3040 (3 )、 JA3036 (4)、 JA3023 (5 ) 和 JA3018 (6)对 于肝癌细胞 HepG-2的抑制作用试验结果的柱状图， 以 5-Fu作阳性对照。  2 is a diagram showing the phenylpiperazine derivative compounds JA3031 (1), JA3024 (2), JA3040 (3 ), JA3036 (4), JA3023 (5 ) and JA3018 (6) provided according to an embodiment of the present invention. A histogram of the inhibition test results of HepG-2 cells, using 5-Fu as a positive control.
图 3为描述根据本发明的实施方案所提供的苯基哌嗪衍生物化合物 JA3031 ( 1 )、 JA3024 (2)、 JA3008 (7)、 JA3031 (8)、 JA3104 (9)对于肝癌细胞 HepG-2 的抑制作用试验结果的柱状图， 以 5-Fu作阳性对照。  3 is a diagram showing the phenylpiperazine derivative compounds JA3031 (1), JA3024 (2), JA3008 (7), JA3031 (8), JA3104 (9) for hepatoma cells HepG-2 according to an embodiment of the present invention. A histogram of the inhibition test results, using 5-Fu as a positive control.
图 4 为描述根据本发明的实施方案所提供的苯基哌嗪衍生物 JA303 JA3024、 JA3036单用及联合用药对于小鼠 S-180肿瘤生长的抑制作用试验结果 的柱状图。 具体实 ifc¾r式  Fig. 4 is a bar graph depicting the results of an experiment for inhibiting the growth of mouse S-180 tumors by the phenylpiperazine derivatives JA303 JA3024, JA3036 alone or in combination according to an embodiment of the present invention. Specific real ifc3⁄4r
下面结合实施例和附图对本发明作进一步详细的描述， 但本发明的实施方 式不限于此。  The present invention will be further described in detail below with reference to the embodiments and drawings, but the embodiments of the present invention are not limited thereto.
实施例 1  Example 1
中间体 1-(4-甲氧基苯基)哌嗪的合成  Synthesis of intermediate 1-(4-methoxyphenyl) piperazine
搅拌下将用氯仿稀释的二乙醇胺的混合液缓慢滴加到冷却至 o°c左右的氯 化亚砜 （与二乙醇胺的摩尔比为 1:4) 与氯仿的混合溶液中， 滴加完毕后， 室温 条件下继续搅拌反应 2-5 h， 然后逐渐升温至 30-70 °C， 维持此温反应 2 h， 反应 完毕后， 加入无水乙醇， 冷却， 减压过滤， 固体分别用乙醇、 ***洗涤， 干燥 得到双 (2-氯乙基)胺盐酸， 产率 97.8%， mp:205.1〜207.0°C。 The mixture of diethanolamine diluted with chloroform was slowly added dropwise to a mixed solution of thionyl chloride (1:4 molar ratio with diethanolamine) and chloroform cooled to about o°C, and the mixture was added dropwise. , room temperature The reaction was stirred for 2-5 h, then gradually warmed to 30-70 ° C, and the temperature was maintained for 2 h. After the reaction was completed, anhydrous ethanol was added, cooled, filtered under reduced pressure, and the solid was washed with ethanol and diethyl ether, respectively. Drying gave bis(2-chloroethyl)amine hydrochloride in a yield of 97.8%, mp: 205.1 to 207.0 °C.
将 4.3 g (24 mmol) 双 (2-氯乙基)胺盐酸盐与 2.5 g (20 mmol) 4-甲氧基苯 胺固体加入到 50 mL正丁醇搅拌混合均匀， 195w功率下微波辐射 6 min， 稍冷 却后加入 1.3g (12 mmol)无水碳酸钠粉末， 继续微波辐射 19 min， 趁热过滤， 少 量热的正丁醇洗涤滤饼， 滤液冷却至室温， 加入一倍体积的无水乙醇混匀后静 置。 过滤所得晶体分别用无水乙醇及***洗涤。 乙醇 /*** （1:2) 重结晶， 真空 干燥得白色晶体 3.2 g，产率 76.9%， mp: 203.6〜205.2°C。 UV (λ/nm) max=246; HPLC (min) Rt=2.717; ^-NMRCDMSO-eD, 300MHz, δ/ppm) 7.03〜6.89(m, 4H, Aryl-H), 4.69(s, 1H, N-H), 3.70 (s, 3H,CH30), 3.32〜3.32(t, 4H, NCH2)， 3.25〜 3.22(t, 4H, NCH2); 13C-NMR(DMSO-6D, 300MHz, δ/ppm) 154.6(C8), 143.6(C5), 119.8(C6, 10), 114.9(C7, 9), 55.6(C11), 47.7(C1, 4), 43.2(C2, 3) 实施例 2 Add 4.3 g (24 mmol) of bis(2-chloroethyl)amine hydrochloride to 2.5 g (20 mmol) of 4-methoxyaniline solids to 50 mL of n-butanol and mix well. Microwave irradiation at 195w power 6 Min, after cooling, add 1.3g (12 mmol) of anhydrous sodium carbonate powder, continue microwave irradiation for 19 min, filter while hot, wash the filter cake with a small amount of hot n-butanol, cool the filtrate to room temperature, add one volume of anhydrous The ethanol was mixed and allowed to stand. The crystals obtained by filtration were washed with absolute ethanol and diethyl ether, respectively. Ethanol/diethyl ether (1:2) was recrystallized, dried in vacuo to give white crystals (yield: </ RTI></RTI></RTI><RTIgt; UV (λ/nm) max=246; HPLC (min) Rt=2.717; ^-NMRCDMSO-eD, 300MHz, δ/ppm) 7.03~6.89(m, 4H, Aryl-H), 4.69(s, 1H, NH ), 3.70 (s, 3H, CH30), 3.32~3.32 (t, 4H, NCH 2 ), 3.25~ 3.22 (t, 4H, NCH 2 ) ; 13 C-NMR (DMSO-6D, 300MHz, δ/ppm) 154.6 (C8), 143.6 (C5), 119.8 (C6, 10), 114.9 (C7, 9), 55.6 (C11), 47.7 (C1, 4), 43.2 (C2, 3) Example 2
3-苯基环戊二酸酑的合成  Synthesis of 3-phenylcyclopentanoate
将 10.6 g苯甲醛（ O.lmol), 26.0 mL(0.2 mol)乙酰乙酸乙酯， 100 mL95%乙 醇混合均匀， 室温搅拌下向上述混合液中缓慢滴加 4.0 mL六氢吡啶溶液， 搅拌 反应 l h〜3 h。 减压过滤， 滤饼用无水乙醇洗涤三次， 所得白色沉淀用无水乙醇 重结晶， 真空干燥， 得白色针状结晶 32.6 g， 即为 2,4-二乙酰基 -3-苯基戊二酸二 乙酯， 产率 93.5% ， 熔点 155.1〜156.9°C。  10.6 g of benzaldehyde (O.lmol), 26.0 mL (0.2 mol) of ethyl acetoacetate, 100 mL of 95% ethanol were uniformly mixed, and 4.0 mL of a solution of hexahydropyridine was slowly added dropwise to the above mixture under stirring at room temperature, and the reaction was stirred for 1 hour. ~3 h. Filtration under reduced pressure, the filter cake was washed three times with anhydrous ethanol, and the obtained white precipitate was recrystallized from anhydrous ethanol and dried in vacuo to give white crystals of crystals of 32.6 g, which is 2,4-diacetyl-3-phenylpentane Diethyl acid, yield 93.5%, melting point 155.1~156.9 °C.
搅拌下将 32.6 g (0.094 mol) 2， 4-二乙酰基 -3-苯基戊二酸二乙酯分批加入 到 100 mL 50%KOH溶液中， 控制温度 30〜80°C之间反应 2小时， 停止加热， 冷却至室温， 冰水浴下向用浓盐酸调节反应液中至 pH约等于 2， 固体析出。 减 压抽滤， 所得滤饼用水洗涤至洗液 pH=6， 所得固体用水重结晶， 干燥， 得白色 粉末 17.9 g， 即为 3-苯基戊二酸， 产率 86.2%， mp: 179.5〜181.7°C。  32.6 g (0.094 mol) of diethyl 2,4-diacetyl-3-phenylglutarate was added in portions to 100 mL of 50% KOH solution under stirring to control the reaction between 30 and 80 °C. After the hour, the heating was stopped, and the mixture was cooled to room temperature, and the reaction liquid was adjusted to a pH of about 2 with concentrated hydrochloric acid under ice-water bath to precipitate a solid. The mixture was filtered under reduced pressure, and the obtained cake was washed with water to a washing solution pH=6, and the obtained solid was recrystallized from water and dried to give white powder, 17.9 g, which was 3-phenylglutaric acid, yield 86.2%, mp: 179.5~ 181.7 ° C.
向 100 mL三颈瓶中加入 17.9 g(0.086 mol) 3-苯基戊二酸， 25 mL乙酸酐， 搅拌溶解，控制温度 125°C回流 8小时自然冷却至室温，冰箱中放置至固体析出。 减压过滤， 滤饼用***洗涤， 弃去滤液， 真空干燥得白色片状晶体 12.5 g， 即为 3-苯基环戊二酸酐， 产率 77.3%， mp: 105.1〜107.6°C。 To a 100 mL three-necked flask, 17.9 g (0.086 mol) of 3-phenylglutaric acid and 25 mL of acetic anhydride were added, and the mixture was stirred and dissolved. The temperature was controlled at 125 ° C for 8 hours, and the mixture was naturally cooled to room temperature, and placed in a refrigerator to precipitate a solid. Filtration under reduced pressure, the filter cake was washed with diethyl ether, the filtrate was discarded, and dried in vacuo to give white crystals of 12.5 g. 3-phenylcycloglutaric anhydride, yield 77.3%, mp: 105.1 to 107.6 °C.
3- (4-甲氧基苯基）环戊二酸酑的合成  Synthesis of 3-(4-methoxyphenyl)cyclopentanoate
将 13.6 g 茴香醛（ O.lmol), 26.0 mL(0.2 mol)乙酰乙酸乙酯， 100 mL95%乙 醇混合均匀， 搅拌下向上述混合液中缓慢滴加 4.0 mL六氢吡啶溶液， 30°C水浴 下反应 1.5 h后析出大量固体， 继续搅拌 2 h。 减压过滤， 所得滤饼用无水乙醇 洗涤三次，所得白色沉淀用 95%乙醇重结晶，真空干燥，得白色针状结晶 35.7 g， 即为 2, 4-二乙酰基 -3- ( 4-甲氧基苯基） 戊二酸二乙酯产率 94.0%， 熔点 148.0〜151.8°C。  13.6 g of anisaldehyde (O.lmol), 26.0 mL (0.2 mol) of ethyl acetoacetate, 100 mL of 95% ethanol were uniformly mixed, and 4.0 mL of a solution of hexahydropyridine was slowly added dropwise to the above mixture under stirring, and a water bath at 30 ° C was added. After 1.5 h of reaction, a large amount of solid precipitated and stirring was continued for 2 h. Filtration under reduced pressure, the obtained filter cake was washed three times with anhydrous ethanol, and the obtained white precipitate was recrystallized from 95% ethanol and dried in vacuo to give white crystals of 35.7 g, which is 2, 4-diacetyl-3- (4- The yield of diethyl methoxyphenyl) glutarate was 94.0%, and the melting point was 148.0 to 151.8 °C.
待 100 mL50%KOH碱溶液温度降至 55°C以下后， 分批加入 35.7 g ( (0.094 mol)) 2， 4-二乙酰基 -3-苯基戊二酸二乙酯， 控制温度 55〜60°C下反应 2小时， 停止加热， 冷却至室温， 冰水浴下向反应液中滴加浓盐酸至 pH=2， 大量固体析 出。 移去冰水浴， 冷却。 减压抽滤， 所得滤饼用水洗涤至洗液 pH=6， 滤饼用蒸 馏水重结晶， 真空干燥， 得白色粉末 21.0 g， 即为 3- (4-甲氧基） 苯基戊二酸， 产率 86%， mp: 181.3〜182.9°C。  After the temperature of 100 mL of 50% KOH alkali solution drops below 55 °C, 35.7 g ((0.094 mol)) of diethyl 2,4-diacetyl-3-phenylglutarate is added in batches to control the temperature of 55~ The reaction was carried out at 60 ° C for 2 hours, the heating was stopped, and the mixture was cooled to room temperature. Concentrated hydrochloric acid was added dropwise to the reaction mixture under ice-water bath to pH = 2, and a large amount of solid was precipitated. Remove the ice water bath and cool. The mixture was filtered under reduced pressure, and the obtained cake was washed with water to a washing solution pH=6. The filter cake was recrystallized from distilled water and dried in vacuo to give a white powder, 21.0 g, which is 3-(4-methoxy)phenylglutaric acid. Yield 86%, mp: 181.3~182.9 °C.
向 100 mL三颈瓶中加入 21.0 g(0.088 mol) 3-(4-甲氧基苯基）戊二酸， 25 mL 乙酸酐， 搅拌溶解， 控制温度 125°C回流 8小时自然冷却至室温， 冰箱中放置， 固体析出。 减压抽滤所得滤饼用***洗涤， 弃去滤液， 真空干燥得白色片状晶 体 13.1 g，即为 3- (4-甲氧基苯基）环戊二酸酐，产率 66.9%, mp: 143.0〜144.6°C。  Add 21.0 g (0.088 mol) of 3-(4-methoxyphenyl)glutaric acid, 25 mL of acetic anhydride to a 100 mL three-necked flask, stir and dissolve, and control to a temperature of 125 ° C for 8 hours to cool to room temperature. Place in the refrigerator and precipitate solids. The filter cake obtained by suction filtration under reduced pressure was washed with diethyl ether, and the filtrate was evaporated, and dried in vacuo to give white crystals of 13.1 g, which was 3-(4-methoxyphenyl)cyclopentanedianhydride, yield 66.9%, mp: 143.0~144.6 °C.
UV (λ/nm): max=227; HPLC (min): Rt=3.159; 3.702； IR (KBr, σ/cm"1): 3433(w, QH), 2966(S, vas CH3; s, vas CH2), 2842(s, vas CH3; vas CH2), 2038〜1942(w, γ -Η), 1808, 1756(vs, carboxylic acid, vc=0), 1516(s, vc=c), 1464(s, Sas CH3; Sas CH2), 1377(s, Sas CH3; Sas CH2), 1254(vs, vas=coc-), 1025(s, vs=coc-), 948(s, γΟΗ), 948〜845(m, V^.H)； 1H-NMR(CH3C1-6D, 300MHz, δ/ppm) 7.14 〜 6.91(m, 4H, Aryl-H), 3.82(s, 3H,CH30), 3.42〜3.36(m, 1H, Ar-CH),3.12〜2.80 (m, 4H, COCH2)， 3.25〜3.22.(t, 4H, NCH2) ; 13C-NMR(CH3C1-6D, 300MHz, δ/ppm) ： 165.8(C2, 3), 159.3(C9), 130.99(C6), 127.2(C7, 11), 114.7(C8, 10), 55.3(C12), 37.4(C1, 5), 33.3(C5)。 实施例 3 UV (λ/nm): max=227; HPLC (min): Rt=3.159; 3.702; IR (KBr, σ/cm" 1 ): 3433 (w, QH), 2966 (S, v as CH3 ; s, v as CH2 ), 2842(s, v as CH3; v as CH2 ), 2038~1942(w, γ -Η ), 1808, 1756(vs, carboxylic acid, v c=0 ), 1516(s, v c =c ), 1464(s, S as CH3 ; S as CH2 ), 1377(s, S as CH3 ; S as CH2 ), 1254(vs, v as = coc- ), 1025(s, v s = coc- ), 948(s, γ ΟΗ ), 948~845(m, V^.H); 1 H-NMR (CH 3 C1-6D, 300MHz, δ/ppm) 7.14 ～ 6.91(m, 4H, Aryl-H ), 3.82 (s, 3H, CH30), 3.42~3.36 (m, 1H, Ar-CH), 3.12~2.80 (m, 4H, COCH 2), 3.25~3.22 (t, 4H, NCH 2);. 13 C-NMR (CH 3 C1-6D, 300MHz, δ/ppm) : 165.8 (C2, 3), 159.3 (C9), 130.99 (C6), 127.2 (C7, 11), 114.7 (C8, 10), 55.3 ( C12), 37.4 (C1, 5), 33.3 (C5). Example 3
5-氧 -3-(4-甲氧基苯基) -5-(4-甲苯基哌嗪)戊酸（编号为 JA3031) 的合成 于 250 mL单口瓶内依次加入 1.0 g (5 mmol) 1-(4-甲基苯基)哌嗪盐酸盐、 1.1 g (5 mmol) 3- (4-甲氧基苯基） 环戊二酸酐、 0.5 mL吡啶、 15 mL无水乙 醇， 130 w功率下微波辐射反应 25-30min， TLC监测至反应完全 (石油醚： 乙酸 乙酯： 甲醇： 冰乙酸 =0.5 mL: 0.5 mL: l滴: 1滴)， 趁热过滤， 无水乙醇淋洗， 所 得滤液静置过夜， 析晶。 减压抽滤， 所得滤饼用少量无水乙醇淋洗。 无水乙醇 重结晶，真空干燥，即得一无色针状结晶 1.5 g，产率 65.2%， mp: 157.4〜159.4°C。 UV (λ/nm): max=247; HPLC (min): Rl=6.990; IR (KBr, σ/cm-l)： 3064〜2510(m, O-H, vAr-HB), 3029(s, vasCH3), 2006(s, vasCH2), 291 l(s, vasCH3), 2827(s, vasCH2), 1990〜1868(w,
Figure imgf000011_0001
1515〜 1491 (s, vc=c), 1450(s, 5asCH3), 1433(s, 5asCH2), 1386(s, 5asCH3), 1369(s, 5asCH2), 1287〜1157(m, v-CN-,v-CO-)， 976(s, γΟΗ), 701(vs, γ=0-Η), 976〜622(m, 5^); ^-NMRCDMSO-eD, 300MHz, δ/ppm) ： 12.01 (s, 1H, OH), 7.27〜7.12(m, 4H, Aryl-H), 7.78〜7.02(m, 4H, Aryl-H), 3.56〜2.95(m, 8H, NCH2), 2.85〜2.79(m, 1H, Aryl-CH), 2.77〜2.62(m, 1H, Aryl-CH), 2.55〜2.51(m, 2H, COCH2), 2.18(s, 3H, Aryl-CH3)； 13C-NMR(DMSO-6D, 300MHz, δ/ppm) ： 173.4(C22), 169.4(C12), 149.1(C15), 144.2(C5), 129.8(C7, 9), 128.6(C17, 19), 128.5(C8), 128.0(C16, 20), 126.7(C18), 116.6(C6, 10), 49.6(C1, 4), 49.2(C2, 3), 45.3(C21), 41.3(C14), 38.74(C13), 24.3(C11) 实施例 4 Synthesis of 5-oxo-3-(4-methoxyphenyl)-5-(4-methylphenylpiperazine)pentanoic acid (No. JA3031) 1.0 g (5 mmol) of 1-(4-methylphenyl)piperazine hydrochloride and 1.1 g (5 mmol) of 3-(4-methoxyphenyl)cyclopentane were added to a 250 mL single-mouth bottle. Anhydride, 0.5 mL pyridine, 15 mL absolute ethanol, microwave irradiation at 130 w for 25-30 min, TLC monitored until the reaction was complete (petroleum ether: ethyl acetate: methanol: glacial acetic acid = 0.5 mL: 0.5 mL: 1 drop: 1 drop), filtered while hot, rinsed with absolute ethanol, and the filtrate was allowed to stand overnight and crystallized. The filter cake was filtered with a small amount of anhydrous ethanol. Recrystallization from absolute ethanol and drying in vacuo gave a colorless needle crystal 1.5 g, yield 65.2%, mp: 157.4~159.4 °C. UV (λ/nm): max=247; HPLC (min): Rl=6.990; IR (KBr, σ/cm-1): 3064~2510 (m, OH, vAr-HB), 3029 (s, vasCH 3 ), 2006(s, vasCH 2 ), 291 l(s, vasCH 3 ), 2827(s, vasCH 2 ), 1990~1868(w,
Figure imgf000011_0001
1515~ 1491 (s, v c = c), 1450(s, 5asCH 3 ), 1433(s, 5asCH 2 ), 1386(s, 5asCH 3 ), 1369(s, 5asCH 2 ), 1287~1157(m, v-CN-,v-CO-), 976(s, γ ΟΗ ), 701(vs, γ =0-Η ), 976~622(m, 5^); ^-NMRCDMSO-eD, 300MHz, δ/ Ppm): 12.01 (s, 1H, OH), 7.27~7.12 (m, 4H, Aryl-H), 7.78~7.02 (m, 4H, Aryl-H), 3.56~2.95 (m, 8H, NCH 2 ), 2.85~2.79 (m, 1H, Aryl- CH), 2.77~2.62 (m, 1H, Aryl-CH), 2.55~2.51 (m, 2H, COCH 2), 2.18 (s, 3H, Aryl-CH3); 13 C-NMR (DMSO-6D, 300MHz, δ/ppm): 173.4 (C22), 169.4 (C12), 149.1 (C15), 144.2 (C5), 129.8 (C7, 9), 128.6 (C17, 19), 128.5 (C8), 128.0 (C16, 20), 126.7 (C18), 116.6 (C6, 10), 49.6 (C1, 4), 49.2 (C2, 3), 45.3 (C21), 41.3 (C14), 38.74 (C13 ), 24.3 (C11) Example 4
4-氧 -4-(4-甲基苯基哌嗪)丁酸 (JA3008) 的合成  Synthesis of 4-oxo-4-(4-methylphenylpiperazine)butyric acid (JA3008)
合成方法与实施例 3类似， 反应物为 1-(4-甲基苯基)哌嗪盐酸盐、 丁二酸酐， 产率 80%， mp: 157.4-159.4°C。 υν (λ/ηηι) max=247; HPLC (min): Rt=4.358; IR (KBr, σ/cm"1) ： 3163〜2470(m, v=c-H), 3032(s, vas CH3), 2923(s, vas CH2), 2812(s, vas CH3), 2753(s, vas CH2), 1993〜1870(w, γ=0-Η), 1742(vs, carboxylic acid vc=0), 1521(vs: amide vc=。)， 1519(s, vc=c), 1480(s, 5as CH3), 1447(s, 5as CH2), 1404(s, Sas CH2), 1379(s, Sas CH2), 1236(s, v-ArN-, V.CO-), H96(s, v-CN-), 934(s, γΟΗ), 803(vs, γΒ -Η),1023〜638(ηι, VAT-H);
Figure imgf000011_0002
300MHz, δ/ppm)： 12.03(s, 1H, OH), 7.04〜6.83 (m, 4H Aryl-H), 3.57—2.79 (m, 8H, NCH2), 2.576〜2.412(t, 4H, COCH2), 2.19(s, 3H, Aryl-CH3)； 13C-NMR(DMSO-6D, 300MHz, δ/ppm) ： 174.4(C15), 170.0(C12), 149.2(C5), 129.8(C7, 9), 128.7(C8), 116.6(C6, 10), 49.6(C1, 4), 49.2(C2, 3), 29.3 (C14), 27.8(C13), 20.5(C11)。 实施例 5 The synthesis was carried out in a similar manner to that in Example 3. The reactant was 1-(4-methylphenyl)piperazine hydrochloride, succinic anhydride, yield 80%, mp: 157.4-159.4 °C. Υν (λ/ηηι) max=247; HPLC (min): Rt=4.358; IR (KBr, σ/cm" 1 ) : 3163~2470 (m, v = cH ), 3032 (s, v as CH3 ), 2923(s, v as CH2 ), 2812(s, v as CH3 ), 2753(s, v as CH2 ), 1993~1870(w, γ =0-Η ), 1742(vs, carboxylic acid v c=0 ), 1521(vs : amide v c =.), 1519(s, v c = c ), 1480(s, 5 as CH3 ), 1447(s, 5 as CH2 ), 1404(s, S as CH2 ), 1379(s, S as CH2 ), 1236(s, v -ArN- , V.CO-), H96(s, v -CN -), 934(s, γ ΟΗ ), 803(vs, γΒ -Η ) , 1023~638(ηι, VAT-H);
Figure imgf000011_0002
300MHz, δ/ppm): 12.03(s, 1H, OH), 7.04~6.83 (m, 4H Aryl-H), 3.57—2.79 (m, 8H, NCH 2 ), 2.576~2.412(t, 4H, COCH2) , 2.19(s, 3H, Aryl-CH 3 ); 13 C-NMR (DMSO-6D, 300MHz, δ/ppm) : 174.4 (C15), 170.0 (C12), 149.2 (C5), 129.8 (C7, 9), 128.7 (C8), 116.6 (C6, 10), 49.6 (C1, 4), 49.2 (C2, 3), 29.3 (C14), 27.8 (C13), 20.5 (C11). Example 5
3-(3，5-二甲氧基苯基 )-5-氧 -5-(4-甲基苯基哌嗪)戊酸 (JA3040) 的合成 合成方法与实施例 3类似， 反应物为 1-(4-甲基苯基)哌嗪盐酸盐、 3- (3,5- 二甲氧基苯基） 环戊二酸酐。 产率 73.5%， mp : 128.5-139.0°C ; UV (λ/nm): max=246HPLC (min): Rt=6.746; IR (KBr, σ/cm"1)： 3011〜2490(m, v0-H, v=c-H), 2954(s, vas CH3), 2907(s, vas CH2), 2837(s, vas CH3), 2797(s, vas CH2), 2113〜1853(w, γ^-κ), 1721(vs, carboxylic acid vc=。), 1586(vs, amide vc=o), 1512〜1487 (s, vc=c), 1463(s, Sas CH3), 1437(s, 5as CH2), 1368(s, Sas CH3), 1332(s, Sas CH2), 1159(vs, vas=coc-), 1022(s, va coc-), 1308〜1065(m, v-CN-,v-co-), 934(s, γΟΗ), 820(vs, γ^-κ), 977〜697 (m, VAT-H) ; ^-NMRCDMSO-eD, 300MHz, δ/ppm) ： 12.03(s, 1H, OH), 7.02〜6.27(m, 8H, Aryl-H), 3.69(s, 3H, OCH3), 3.60〜2.59(m, 8H, NCH2), 3.00〜2.96(m, 1H, Aryl-CH) 2.84〜2.53(m, 4H, COCH2), 2.18(s, 3H, Aryl-CH3); 13C-NMR(DMSO-6D, 300MHz, δ/ppm)： 173.4(C24), 169.5 (C12), 160.7(17, 19), 149.2(C5), 146.6(C15), 129.8(C7, 9), 128.7(C8), 116.6(C6, 10), 106.2(C16, 20), 98.3(C18), 56.5(C21, 22), 55.5(C1, 4), 49.6(C2, 3), 49.4(C23), 45.4(C14), 38.6(C13), 20.5(C11)。 实施例 6 The synthesis synthesis method of 3-(3,5-dimethoxyphenyl)-5-oxo-5-(4-methylphenylpiperazine)pentanoic acid (JA3040) is similar to that of Example 3, and the reactant is 1 -(4-Methylphenyl)piperazine hydrochloride, 3-(3,5-dimethoxyphenyl)cycloglutaric anhydride. Yield 73.5%, mp: 128.5-139.0 ° C; UV (λ/nm): max=246 HPLC (min): Rt=6.746; IR (KBr, σ/cm" 1 ): 3011~2490 (m, v 0 -H , v =cH ), 2954(s, v as CH3 ), 2907(s, v as CH2 ), 2837(s, v as CH3 ), 2797(s, v as CH2 ), 2113~1853(w, γ^-κ), 1721(vs, carboxylic acid v c= .), 1586(vs, amide vc=o), 1512~1487 (s, v c=c ), 1463(s, S as CH3 ), 1437 (s, 5 as CH2 ), 1368(s, S as CH3 ), 1332(s, S as CH2 ), 1159(vs, v as = coc- ), 1022(s, v a coc- ), 1308~1065 (m, v -CN- , v -co- ), 934(s, γ ΟΗ ), 820(vs, γ^-κ), 977~697 (m, VAT-H) ; ^-NMRCDMSO-eD, 300MHz , δ/ppm): 12.03(s, 1H, OH), 7.02~6.27(m, 8H, Aryl-H), 3.69(s, 3H, OCH 3 ), 3.60~2.59(m, 8H, NCH 2 ), 3.00~2.96(m, 1H, Aryl-CH) 2.84~2.53(m, 4H, COCH2), 2.18(s, 3H, Aryl-CH 3 ); 13 C-NMR (DMSO-6D, 300MHz, δ/ppm) : 173.4(C24), 169.5 (C12), 160.7(17, 19), 149.2(C5), 146.6(C15), 129.8(C7, 9), 128.7(C8), 116.6(C6, 10), 106.2(C16 , 20), 98.3 (C18), 56.5 (C21, 22), 55.5 (C1, 4), 49.6 (C2, 3), 49.4 (C23), 45.4 (C14), 38.6 (C13), 20.5 (C11). Example 6
3-(4-甲氧苯基) -5-(4-(4-甲氧苯基)哌嗪 -5-氧 -戊酸 (JA3024) 的合成  Synthesis of 3-(4-methoxyphenyl)-5-(4-(4-methoxyphenyl)piperazine-5-oxo-pentanoic acid (JA3024)
合成方法与实施例 3类似， 反应物为 1-(4-甲氧基苯基)哌嗪盐酸盐、 3- (4- 甲氧基苯基）环戊二酸酐。产率 88.1%， mp: 171.4-173.6°C ; UV ( /nm) max=235, λ=290; HPLC (min): Rt=6.977; IR (KBr, σ/cm"1)： 3423(m, v0-H), 3002(s, vas CH3), 2912(s, vas CH2), 2861(s, vas CH3), 2832(s, vas CH2), 2058〜1905(w, y=c-H), 1715(vs, carboxylic acid vc=。), 1609(vs, amide vc=o), 1576〜1492 (s, vc=c), 1461(s, Sas CH3), 1436(s, Sas CH2), 1386(s, Sas CH3), 1368(s, Sas CH2), 1246(vs, va coc-), 1031(s, v coc-), 1329〜l l l l(m, v-CN-,v-co-), 992〜667(m, AT-H) ; ^-NMRCDMSO-eD, 300MHz, δ/ppm)： 11.99(s, 1H, OH), 7.19〜6.79(m, 8H, Aryl-H), 3.66(s, 6H, OCH3), 3.57〜 2.88(m, 8H, NCH2), 2.77〜2.73(m, 1H, Aryl-CH) 2.69〜2.56(m, 4H, COCH2); 13C-NMR(DMSO-6D, 300MHz, δ/ppm) ： 173.4(C23), 169.5(C12), 158.1(C18, 8), 153.8(C5), 136.0(C15), 129.0(C16, 20), 118.6(C7, 9,), 114.7(C17, 19), 114.0(C6, 10), 55.6(C11), 55.4(21), 50.6, 50.4(C1, 4), 45.4(C2, 3), 41.4(C22), 40.7(C14), 39.0(C22),
Figure imgf000013_0001
实施例 7 The synthesis was carried out in the same manner as in Example 3, and the reactant was 1-(4-methoxyphenyl)piperazine hydrochloride, 3-(4-methoxyphenyl)cyclopentanedianhydride. Yield 88.1%, mp: 171.4-173.6 ° C; UV ( /nm) max = 235, λ = 290; HPLC (min): Rt = 6.977; IR (KBr, σ/cm" 1 ): 3423 (m, v 0-H ), 3002(s, v as CH3 ), 2912(s, v as CH2 ), 2861(s, v as CH3 ), 2832(s, v as CH2 ), 2058~1905(w, y = cH ), 1715(vs, carboxylic acid v c= .), 1609(vs, amide vc=o), 1576~1492 (s, v c=c ), 1461(s, S as CH3 ), 1436(s, S as CH2 ), 1386(s, S as CH3 ), 1368(s, S as CH2 ), 1246(vs, v a coc- ), 1031(s, v coc- ), 1329~llll(m, v - CN- , v -co- ), 992~667(m, AT-H) ; ^-NMRCDMSO-eD, 300MHz, δ/ppm): 11.99 (s, 1H, OH), 7.19~6.79 (m, 8H, Aryl-H), 3.66 (s, 6H, OCH 3 ), 3.57~ 2.88 (m, 8H, NCH 2 ), 2.77 ~2.73 (m, 1H, Aryl-CH) 2.69~2.56 (m, 4H, COCH2); 13 C-NMR (DMSO-6D, 300MHz, δ/ppm): 173.4 (C23), 169.5 (C12), 158.1 ( C18, 8), 153.8 (C5), 136.0 (C15), 129.0 (C16, 20), 118.6 (C7, 9,), 114.7 (C17, 19), 114.0 (C6, 10), 55.6 (C11), 55.4 (21), 50.6, 50.4 (C1, 4), 45.4 (C2, 3), 41.4 (C22), 40.7 (C14), 39.0 (C22),
Figure imgf000013_0001
Example 7
5-(4-(4-乙氧甲酰苯基)哌嗪) -3-(4-甲氧基苯基) -5-氧 -3-戊酸 (JA3031) 的合 成  Synthesis of 5-(4-(4-ethoxyformylphenyl)piperazine)-3-(4-methoxyphenyl)-5-oxo-3-pentanoic acid (JA3031)
于 250 mL单口瓶内依次加入 1.4 g (5 mmol) 1 -(4-乙氧基甲酰苯基)哌嗪盐 酸盐， 1.1 g (5 mmol) 3- (4-甲氧基苯基）戊二酸酐、 0.5 mL吡啶、 0.2 g DMAP， 15 mL丙酮， 65 w功率下微波辐射 30min， TLC监测至反应完全 (石油醚： 乙酸 乙酯： 甲醇： 冰乙酸 /0.5 mL: 0.5 mL:l滴: 1滴）， 趁热过滤， 2〜3 mL丙酮淋洗 2 次， 所得滤液静置过夜， 析晶。 减压抽滤， 少量丙酮淋洗， 所得滤饼用无水乙 醇重结晶，真空干燥即得一无色片状结晶 1.2 g， 产率 52.9%, mp: 162.1-163.4°C。 UV (λ/nm)： max=302; HPLC (min) Rt=11.413 ; IR(KBr, σ/cm-l)： 〜3000(m, V=C-H), 1712(vs, carboxylic acid vc=o), 1608, 1578(vs, amide vc=0), 1516(s, vc=c), 1462(s, 5as CH3; 5asCH2), 1390(s, 5asCH3), 1369(s, 5asCH2), 1286(vs, vas=COC-)， 1232(s, vs=coc-), 1187 〜 1027(m, -CN-, v-CO-), 977 〜 655(m, ^-H) ； ^-NMRCDMSO-eD, 300MHz, δ/ppm) ： 11.97(s, 1H, OH), 7.78〜6.79(m, 4H, Aryl-H), 4.25〜4.17(q, 2H, CH3CH2), 3.68(s, 6H, CH30), 3.68〜3.04(m, 8H, NCH2), 2.72〜2.59(m, 4H, COCH2), 2.53〜 2.48 (m, 1H, Aryl-CH), 1.29〜1.25(t, 3H, C¾CH2，） ； 13C-NMR(DMSO-6D, 300MHz, δ/ppm) ： 173.5(C25), 169.7(C14), 166.1(C11), 158.2(C20), 153.9(C5), 136.1(C17), 131.1(C7, 9), 129.0(C6, 10), 128.0(C18, 22), 119.1(C8), 114.0, 113.9(C19, 21), 60.3(C12), 55.3(C23), 47.1(C1, 4), 46.8(C2, 3), 44.8(C24), 40.9(C16), 37.9(C15), 14.7(C 13)。 实施例 8 3-(3，5-二甲氧基苯基 )-5-(4-(4-乙氧甲酰)苯基)哌嗪） -5-氧 -3-戊酸（ JA3023) 合成方法与实施例 7类似， 反应物为 1-(4-乙氧基甲酰苯基)哌嗪盐酸盐、 3- (3,5-二甲氧基苯基）环戊二酸酐。产率 53.2%, mp: 147.4-151.4°C； UV ( /nm): max=231, λ=302; HPLC (min): Rt= 12.224； IR (KBr, σ/cm"1)： 3427(m, v0-H), 2937(s: vAS CH3, vAS CH2, vAS CH3, CH2), 〜1898(w,
Figure imgf000014_0001
1721(VS, carboxylic acid vc=0), 1586(vs amide vc=o), 1516, 1481 (s, vc=c), 1458(s, Sas CH3), 1437(s, Sas CH2), 1389(s, Sas CH3), 1366(s, Sas CH2), 1191(vs, vas=coc-), 1053(s, vs=coc-), 1245〜1024(m, v-CN-, v-CO-), 934(s, γΟΗ), 813(VS,
Figure imgf000014_0002
300MHz, δ/ppm) ： 12. 00(s, 1H, OH), 7.79〜6.27(m, 8H, Aryl-H), 4.25〜4.17(q, 2H, CH3CH2), 3.65(s, 3H,CH30), 3.52〜3.02(m, 8H, NCH2), 2.69〜2.63(m, 4H, COCH2), 2.50—2.48 (m, 1H, Aryl-CH), 1.29〜1.25(t, 3H, CH3CH2，)； 13C-NMR(DMSO-6D, 300MHz, δ/ppm): 173.4(C26), 169.7(C14), 166.1(C11), 160.7(C19, 21), 153.9(C5), 146.7(C17): 131.1(C7, 9), 119.1(C8), 114.0(C6, 10), 106.2(C18, 22), 98.2(C20), 60.3 (C12), 55.5(C23, 24), 47.1 (CI, 4), 46.8(C2, 3), 44.8(C25), 38.9(C16), 38.6(C15), 14.1(C13) 实施例 9 1.4 g (5 mmol) of 1-(4-ethoxyformylphenyl)piperazine hydrochloride, 1.1 g (5 mmol) of 3-(4-methoxyphenyl) was added to a 250 mL single-mouth vial. Glutaric anhydride, 0.5 mL pyridine, 0.2 g DMAP, 15 mL acetone, microwave irradiation at 65 w for 30 min, TLC monitored until the reaction was complete (petroleum ether: ethyl acetate: methanol: glacial acetic acid / 0.5 mL: 0.5 mL: 1 drop : 1 drop), filtered while hot, 2 to 3 mL of acetone was rinsed twice, and the resulting filtrate was allowed to stand overnight and crystallized. The mixture was filtered under reduced pressure with a small amount of acetone. The obtained cake was recrystallized from anhydrous ethanol and dried in vacuo to give a colorless crystals of crystals of 1.2 g, yield 52.9%, mp: 162.1-163.4 °C. UV (λ/nm): max=302; HPLC (min) Rt=11.413; IR (KBr, σ/cm-1): ~3000 (m, V=CH), 1712 (vs, carboxylic acid v c = o ), 1608, 1578(vs, amide v c=0 ), 1516(s, v c =c), 1462(s, 5as CH 3 ; 5asCH 2 ), 1390(s, 5asCH 3 ), 1369(s, 5asCH 2) , 1286(vs, vas=COC-), 1232(s, vs =coc- ), 1187 ~ 1027(m, -CN-, v-CO-), 977 ~ 655(m, ^-H) ; ^-NMRCDMSO-eD, 300MHz, δ/ppm): 11.97(s, 1H, OH), 7.78~6.79 (m, 4H, Aryl-H), 4.25~4.17 (q, 2H, CH 3 CH 2 ), 3.68 (s, 6H, CH 3 0), 3.68~3.04 (m, 8H, NCH 2 ), 2.72~2.59 (m, 4H, COCH 2 ), 2.53~ 2.48 (m, 1H, Aryl-CH), 1.29~1.25 (t, 3H, C3⁄4CH 2 ,) ; 13 C-NMR (DMSO-6D, 300MHz, δ/ppm) : 173.5 (C25), 169.7 (C14), 166.1 (C11), 158.2 (C20), 153.9 (C5) , 136.1(C17), 131.1(C7, 9), 129.0(C6, 10), 128.0(C18, 22), 119.1(C8), 114.0, 113.9(C19, 21), 60.3(C12), 55.3(C23) , 47.1 (C1, 4), 46.8 (C2, 3), 44.8 (C24), 40.9 (C16), 37.9 (C15), 14.7 (C 13). Example 8 Synthesis and implementation of 3-(3,5-dimethoxyphenyl)-5-(4-(4-ethoxycarbonyl)phenyl)piperazine)-5-oxo-3-pentanoic acid (JA3023) Similarly to Example 7, the reactant was 1-(4-ethoxyformylphenyl)piperazine hydrochloride, 3-(3,5-dimethoxyphenyl)cycloglutaric anhydride. Yield 53.2%, mp: 147.4-151.4 ° C; UV ( /nm): max = 231, λ = 302; HPLC (min): Rt = 12.224; IR (KBr, σ/cm" 1 ): 3427 (m , v 0-H ), 2937(s : v AS C H3, v AS C H2, v AS C H3, CH2), ~1898(w,
Figure imgf000014_0001
1721(VS, carboxylic acid v c=0 ), 1586(vs amide vc=o), 1516, 1481 (s, v c=c ), 1458(s, S as CH3 ), 1437(s, S as CH2 ) , 1389(s, S as CH3 ), 1366(s, S as CH2 ), 1191(vs, v as = coc- ), 1053(s, v s = coc- ), 1245~1024(m, v -CN -, v -CO -), 934(s, γ ΟΗ ), 813 (VS,
Figure imgf000014_0002
300MHz, δ/ppm): 12. 00(s, 1H, OH), 7.79~6.27(m, 8H, Aryl-H), 4.25~4.17(q, 2H, CH 3 CH 2 ), 3.65(s, 3H ,CH30), 3.52~3.02(m, 8H, NCH 2 ), 2.69~2.63(m, 4H, COCH2), 2.50—2.48 (m, 1H, Aryl-CH), 1.29~1.25(t, 3H, CH 3 CH 2 ,); 13 C-NMR (DMSO-6D, 300MHz, δ/ppm): 173.4 (C26), 169.7 (C14), 166.1 (C11), 160.7 (C19, 21), 153.9 (C5), 146.7 ( C17) : 131.1 (C7, 9), 119.1 (C8), 114.0 (C6, 10), 106.2 (C18, 22), 98.2 (C20), 60.3 (C12), 55.5 (C23, 24), 47.1 (CI, 4), 46.8 (C2, 3), 44.8 (C25), 38.9 (C16), 38.6 (C15), 14.1 (C13) Example 9
3-(4-氯苯基 )- 5-(4-(4-甲氧基苯基)哌嗪） -5-氧 -戊酸 (JA3036) 的合成  Synthesis of 3-(4-chlorophenyl)-5-(4-(4-methoxyphenyl)piperazine)-5-oxo-pentanoic acid (JA3036)
合成方法与实施例 7类似， 反应物为 1-(4-甲氧基苯基)哌嗪盐酸盐、 3- (4- 氯苯基）环戊二酸酐。 产率 65.7%， mp: 203.2-206.5 °C ; UV (λ/nm): max=228; HPLC (min): Rt=4.665; IR (KBr, σ/cm"1): 〜3000(m, 2837(m, v=c-H), 〜1906(w, 1716(VS, carboxylic acid, vc=o), 1583(vs, amide vc=o), 1516, 1481 (s, vc=c), 1494(s, B 5as CH3; Sas CH2), 〜1396(s, Sas CH3; Sas CH2), 1234(vs, vas=coc-), 1158(s, vs =coc-), 1285〜1030(m, v-CN-, v-co-), 981〜665(m, AT-H)； ^-NMRCDMSO-eD, 300MHz, δ/ppm): 11.97(s, 1H, OH), 7.24〜7.20(m, 2H, Aryl-H), 7.18〜7.16 (m, 2H, Aryl-H), 6.92〜6.88 (m, 2H, Aryl-H), 6.81〜6.79(m, 2H, Aryl-H), 3,66(s, 3H, CH30), 3.57〜2.87(m, 8H, NCH2), 2.81〜2.76(m, 1H, Aryl-CH), 2.69〜2.59(m, 2H, COCH2), 2.51〜2.42(m, 2H, COCH2); 13C-NMR(DMSO-6D, 300MHz, δ/ppm): 173.3(C22), 169.6(C11), 158.1(C17), 150.0(C5), 136.0(C14), 129.8(C7, 9), 128.9 (C15, 19), 123.2(C8), 117.6(C6, 10), 113.9(C16, 18), 55.3(C20), 48.7(C1, 4), 48.5(C2, 3), 45.1(C21), 38.0(C13), 31.1(C12)。 实施例 10 The synthesis was carried out in the same manner as in Example 7, and the reactant was 1-(4-methoxyphenyl)piperazine hydrochloride, 3-(4-chlorophenyl)cyclopentanedianhydride. Yield 65.7%, mp: 203.2-206.5 °C; UV (λ/nm): max=228; HPLC (min): Rt=4.665; IR (KBr, σ/cm" 1 ): ~3000 (m, 2837) (m, v = cH ), ~1906 (w, 1716 (VS, carboxylic acid, v c = o), 1583 (vs, amide vc=o), 1516, 1481 (s, v c = c), 1494 ( s, B 5 as CH3 ; S as CH2 ), ~1396(s, S as CH3 ; S as CH2 ), 1234(vs, v as = coc- ), 1158(s, v s =coc- ), 1285~ 1030(m, v -CN- , v -co- ), 981~665(m, AT-H); ^-NMRCDMSO-eD, 300MHz, δ/ppm): 11.97(s, 1H, OH), 7.24~ 7.20(m, 2H, Aryl-H), 7.18~7.16 (m, 2H, Aryl-H), 6.92~6.88 (m, 2H, Aryl-H), 6.81~6.79 (m, 2H, Aryl-H), 3,66(s, 3H, CH 3 0), 3.57~2.87(m, 8H, NCH 2 ), 2.81~2.76(m, 1H, Aryl-CH), 2.69~2.59(m, 2H, COCH2), 2.51 ~2.42 (m, 2H, COCH2); 13 C-NMR (DMSO-6D, 300MHz, δ/ppm): 173.3 (C22), 169.6 (C11), 158.1 (C17), 150.0 (C5), 136.0 (C14) , 129.8(C7, 9), 128.9 (C15, 19), 123.2 (C8), 117.6 (C6, 10), 113.9 (C16, 18), 55.3 (C20), 48.7 (C1, 4), 48.5 (C2, 3), 45.1 (C21), 38.0 (C13), 31.1 (C12). Example 10
3- (3，5-二甲氧基苯基 )-5-(4-苄基哌嗪） -5-氧 -3-戊酸（JA3018) 的合成  Synthesis of 3-(3,5-dimethoxyphenyl)-5-(4-benzylpiperazine)-5-oxo-3-pentanoic acid (JA3018)
合成反应同实施例 7， 反应物为 1- (苄基基)哌嗪盐酸盐、 3- (3,5-二甲氧基苯 基）环戊二酸酐。 产率 65.6%; UV( / nm): 279.00(W), 211.03( M); 207.03(M)， 226.02( VS); IR(KBr, σ /cm"1): 3061.41， 3018.50 (v =C-H)， 2939.13， 2838.40 (v C-H)， 1701.26， 1630.31 (v c=0)， 1467.08, 1511.86 (v c=c)， 743.44， 701.71 (5=C-H)； iH-NMRpOO MHz, NaOH, D20)S/ppm ： 7.253〜7.194(2H， H-9， 13， m) ， 7.104〜7.033 (3H， H-10, H-ll , H-12, m)， 6.784〜6.770(3H， H-19, H-23 , H-21 , m)， 3.666(6H， H-29, H-30, s)， 3.510〜3.532(1H， H-17, m)， 3.285〜 3.264(1H， H-7b， d)， 3.225〜3.193(2H， H-3b， H-5b， t)， 3.137〜3.116(1H， H-7a， d)， 3.089〜3.055(2H， H-3b， H-5b， t)， 2.936〜2.576(2H， H-3a， H-5a， t)， 2.433〜 2.360(2H, H-16, m)， 2.325〜2.305(2H， H-2b， H-6b， m) ， 2.148〜2.129(2H， H-2a， H-6a， d)， 1.838〜 1.808(1H， H-24b， s)， 1.206(1H， H -24a , s) ； 13C-NMR(300MHz， NaOH， D20)S/ppm： 180.639(C-25) 172.759(C-14)， 157.843(C-20 , C-22)， 135.596(C-18)， 130.238(C-8)， 128.917(C-9 , C-13)， 128.650(C-10， C-12) , 127.997(C-11) , 114.254(C-19， C-23)， 62.028(C-21) , 55.423(C-7), 51.891(C-2， C-6)， 51.520(C-29, C-30), 45.658(C-24), 44.218 (C-16)， 41.250(C-17), 40.300(C-3)， 39.113(C-5) 。 The synthesis was carried out in the same manner as in Example 7, and the reactant was 1-(benzyl)piperazine hydrochloride, 3-(3,5-dimethoxyphenyl)cyclopentanedianhydride. Yield 65.6%; UV ( / nm): 279.00 (W), 211.03 (M); 207.03 (M), 226.02 (VS); IR (KBr, σ /cm" 1 ): 3061.41, 3018.50 (v =CH ) , 2939.13, 2838.40 (v CH ), 1701.26, 1630.31 (v c=0 ), 1467.08, 1511.86 (v c=c ), 743.44, 701.71 (5 =CH ); iH-NMRpOO MHz, NaOH, D 2 0)S /ppm: 7.253~7.194(2H, H-9, 13, m), 7.104~7.033 (3H, H-10, H-ll, H-12, m), 6.784~6.770 (3H, H-19, H -23, H-21, m), 3.666 (6H, H-29, H-30, s), 3.510~3.532 (1H, H-17, m), 3.285~ 3.264 (1H, H-7b, d) , 3.225~3.193(2H, H-3b, H-5b, t), 3.137~3.116(1H, H-7a, d), 3.089~3.055(2H, H-3b, H-5b, t), 2.936~ 2.576(2H, H-3a, H-5a, t), 2.433~ 2.360(2H, H-16, m), 2.325~2.305(2H, H-2b, H-6b, m), 2.148~2.129 (2H , H-2a, H-6a, d), 1.838~ 1.808(1H, H-24b, s), 1.206(1H, H -24a, s) ; 13 C-NMR (300MHz, NaOH, D 2 0)S /ppm: 180.639(C-25) 172.759(C-14), 157.843(C-20, C-22), 135.596(C-18), 130.238(C-8), 128.917(C-9, C-13 ) 128.650 (C-10, C-12), 127.997 (C-11), 114.254 (C-19, C-23), 62.028 (C-21), 55.423 (C-7), 51.891 (C-2, C -6), 51.520 (C-29, C-30), 45.658 (C-24), 44.218 (C-16), 41.250 (C-17), 40.300 (C-3), 39.113 (C-5).
实施例 11  Example 11
4- (4-甲苯磺難哌嗪)苯甲酸乙酯（JA3104) 的合成  Synthesis of ethyl 4-(4-toluenesulfonazine) benzoate (JA3104)
将 1.4 g (5 mmol) 1-(4乙氧甲酰苯基)哌嗪加入到 20 mL吡啶中， 室温搅 拌下分批加入对甲苯磺酰氯（数量， mol)， 加完后控温（30〜50°C )反应？ 小时。 冷却， 过滤， 反应液缓慢倒入到 lOOmL冰水中， 搅拌 3h后过滤， 所得白色固 体以无水乙醇重结晶，真空干燥得白色针状细晶 1.2 g，产率 62.9%, mp: 132.9〜 134.9°C。 UV (λ/nm)： max=299, λ=229; IR(KBr, σ/cm"1) 3049〜2614(m, v=c-H), 2978(s, vas CH3; vas CH2), 2840 (s, vas CH3; Bvs CH2), 1924(w,
Figure imgf000015_0001
1706(VS, carboxylic acid vc=0), 1579(s, vc=c), 1450(s, Sas CH3), 1390(s, Sas CH2), 1343(s, Sas CH3), 1311(s, Sas CH2), 1283(vs, vas SO2-N) , 1236 (vs, vas=coc-), 1188(s, vs=coc-), 1163(vs, vas S02-N), 942(s, γΟΗ), 736(VS, γ=0-Η), 942〜648(m, AT-H); ^-NMRCDMSO-eD, 300MHz, δ/ppm) 7.75〜6.92(m, 4H, Aryl-H), 4.24〜4.17(q, 2H, CH3CH2), 3.41〜2.94(m, 8H, NCH2), 2.38(s, 3H, OCH3), 1.27〜1.24(t, 3H, CH3CH2) ； 13C-NMR(DMSO-6D, 300MHz, δ/ppm) 165.9(C11), 153.5(C5), 144.4(C17), 132.0(C14), 131.1(C7, 9), 130.3(C16, 18), 128.1(C15, 19), 119.7(C8), 114.4(C6, 10), 60.4(C12), 46.7(C1, 4), 45.9(C2, 3), 21.4(C20), 14.7(C13)。 实施例 12 Add 1.4 g (5 mmol) of 1-(4 ethoxycarbonylphenyl) piperazine to 20 mL of pyridine, and add p-toluenesulfonyl chloride (quantity, mol) in portions at room temperature with stirring. ~50 ° C) reaction? hour. After cooling, filtration, the reaction solution was poured slowly into 100 mL of ice water, stirred for 3 hours, and filtered. The obtained white solid was recrystallized from anhydrous ethanol, and dried in vacuo to give white needles fine crystals of 1.2 g, yield 62.9%, mp: 132.9~ 134.9 °C. UV (λ/nm): max=299, λ=229; IR(KBr, σ/cm" 1 ) 3049~2614(m, v =cH ), 2978(s, v as CH3 ; v as CH2 ), 2840 (s, v as CH3 ; Bv s CH2 ), 1924(w,
Figure imgf000015_0001
1706 (VS, carboxylic Acid v c = 0 ), 1579(s, v c = c ), 1450(s, S as CH3 ), 1390(s, S as CH2 ), 1343(s, S as CH3 ), 1311(s, S as CH2 ), 1283(vs, v as SO2-N) , 1236 (vs, v as = coc- ), 1188(s, v s = coc- ), 1163(vs, v as S02-N ), 942(s , γ ΟΗ ), 736 (VS, γ =0-Η ), 942~648 (m, AT-H); ^-NMRCDMSO-eD, 300MHz, δ/ppm) 7.75~6.92(m, 4H, Aryl-H ), 4.24~4.17(q, 2H, CH 3 CH 2 ), 3.41~2.94(m, 8H, NCH 2 ), 2.38(s, 3H, OCH3), 1.27~1.24(t, 3H, CH 3 CH 2 ) 13 C-NMR (DMSO-6D, 300MHz, δ/ppm) 165.9 (C11), 153.5 (C5), 144.4 (C17), 132.0 (C14), 131.1 (C7, 9), 130.3 (C16, 18), 128.1 (C15, 19), 119.7 (C8), 114.4 (C6, 10), 60.4 (C12), 46.7 (C1, 4), 45.9 (C2, 3), 21.4 (C20), 14.7 (C13). Example 12
苯基哌嗪衍生物对人脐静脉内皮细胞 (ECV304) 的抑制活性  Inhibitory activity of phenylpiperazine derivatives on human umbilical vein endothelial cells (ECV304)
选用肿瘤血管内皮细胞相当的人脐静脉内皮细胞 (ECV304)增殖的药理模型 对目标产物进行初步筛选。 MTT还原法来检测细胞增殖及凋亡。  A pharmacological model of human umbilical vein endothelial cell (ECV304) proliferation comparable to tumor vascular endothelial cells was selected for preliminary screening of the target product. MTT reduction method was used to detect cell proliferation and apoptosis.
具体试验方法：  Specific test methods:
实验样品： 分别选取苯基哌嗪化合物 5-氧 -3-苯基 -5-(4-甲苯基哌嗪)戊酸 (JA3031 , 见实施例 3， 编号为 1 ); 3-(4-甲氧苯基) -5-(4-(4-甲氧苯基)哌嗪 -5-氧 -戊酸 （JA3024, 见实施例 6， 编号为 2); 3-(3, 5-二甲氧基苯基 )-5-氧 -5-(4-对甲 苯基哌嗪)戊酸（JA3040, 见实施例 5， 编号为 3 ) 以及 4-氧 -4-(4-对甲苯基哌嗪) 丁酸 (JA3008, 见实施例 4， 编号为 4)。  Experimental sample: phenyl piperazine compound 5-oxo-3-phenyl-5-(4-methylphenylpiperazine) valeric acid (JA3031, see Example 3, No. 1); 3-(4-A Oxyphenyl) -5-(4-(4-methoxyphenyl)piperazine-5-oxo-valeric acid (JA3024, see Example 6, number 2); 3-(3, 5-dimethoxy Phenyl)-5-oxo-5-(4-p-tolylpiperazine) valeric acid (JA3040, see Example 5, number 3) and 4-oxo-4-(4-p-tolylpiperazine) Butyric acid (JA3008, see Example 4, number 4).
样品液配置： 按照化合物预实验结果 22.5 mol/L 的样品活性抑制率达 60% 左右， 试验设计将所配溶液浓度集中在预期抑制率达 50%所需化合物的即 30μηιο1 左右， 其余较低浓度作为对照， 即 200 μηιοΙ/L的原液用培养基分别按 照 1： 4， 1： 8， 1： 40， 1： 200， 1： 2000稀释得到所需的 5个浓度梯度： 5(^mol/L、 25 mol/L、 5 mol/L、 Ιμηιοΐ/L, 0.1 mol/L。  Sample solution configuration: According to the pre-experiment of the compound, the activity inhibition rate of the sample of 22.5 mol/L is about 60%. The experimental design concentrates the concentration of the solution in the desired compound with a desired inhibition rate of 50%, that is, 30μηιο1, and the other lower concentration. As a control, the stock solution of 200 μηιοΙ/L was diluted with 1:4, 1:8, 1:40, 1:200, 1:2000 to obtain the desired five concentration gradients: 5 (^mol/L) 25 mol/L, 5 mol/L, Ιμηιοΐ/L, 0.1 mol/L.
对照设计： 5(^mol/L 的 5-Fu (50μηιο1/Ε) 为阳性对照， 1.25%DMSO为空 白对照， 为了避免 MTT及环境因素造成的干扰， 每个浓度梯度设置 8个复空， 尽量减小误差。  Control design: 5 (^mol/L 5-Fu (50μηιο1/Ε) as a positive control, 1.25% DMSO as a blank control, in order to avoid interference caused by MTT and environmental factors, set 8 complexes for each concentration gradient, try to Reduce the error.
细胞培养 将 ECV304冻存管从液氮罐中取出， 迅速放入 37°C温水中， 5分 钟后在无菌操作台上， 将冻存管放入离心管， 离心 (1000r/min)5分钟， 吸去上层 液体，加入适量无血清 RPMI-1640培养液，用吸管将细胞吹散，离心 (1000r/min)5 分钟，吸去上层液体。在细胞中加入含 10%胎牛血清、 100U/ml青霉素与 10(^g/ml 链霉素的 RPMI-1640培养液， 用吸管将细胞吹散， 接种入 50ml培养瓶。 37°C， 5% 02培养箱中培养， 隔天换液。 第四天开始有细胞融合， 形态为鹅卵石形状， 传代培养。 Cell culture The ECV304 cryotube was taken out from the liquid nitrogen tank and quickly placed in warm water at 37 ° C. After 5 minutes, the cryotube was placed in a centrifuge tube and centrifuged (1000 r/min) for 5 minutes. , suck the upper layer For the liquid, add an appropriate amount of serum-free RPMI-1640 medium, blow the cells with a pipette, centrifuge (1000r/min) for 5 minutes, and aspirate the supernatant. RPMI-1640 medium containing 10% fetal calf serum, 100 U/ml penicillin and 10 (^g/ml streptomycin) was added to the cells, and the cells were blown off with a pipette and inoculated into a 50 ml culture flask. 37 ° C, 5 The culture was carried out in an incubator of % 0 2 and changed every other day. On the fourth day, cell fusion was carried out, and the shape was cobblestone shape, subcultured.
种板：细胞分别用胰酶消化、含 10%胎牛血清 RPMI-1640培养液调成悬液， 以每孔 ΙΟΟμΙ (104个细胞)接种于 96孔培养板中，在 37°C、 5%C02培养箱中培养 至细胞贴壁。 Seed plate: The cells were separately diluted with trypsin, containing 10% fetal bovine serum RPMI-1640 medium, and inoculated into 96-well culture plates at 37 ° C, 5 μM per well (10 4 cells). Incubate in the %C0 2 incubator until the cells are attached.
加样： 加入待测样品 （100μΙ7孔）。 放入 37°C培养箱中培育 72h。  Loading: Add the sample to be tested (100 μΙ 7 wells). Incubate in a 37 ° C incubator for 72 h.
显色：每孔加 20μ1 ΜΤΤ染液 (5mg/mL的 PBS液，滤过除菌， 4°C避光保存)， 培养箱中继续培育 4h。  Color development: Add 20 μl of sputum solution (5 mg/mL PBS solution per well, filter and sterilize, store at 4 °C in the dark), and continue incubation for 4 hours in the incubator.
测试： 甩尽细胞培养液， 每孔加入 10(^L DMSO， 振摇 10min， 待甲臜充分 溶解后， 酶标仪测 OD值。  Test: Drain the cell culture medium, add 10 (^L DMSO per well, shake for 10 min, and wait until the formazan is fully dissolved, and measure the OD value by the microplate reader.
结果显示苯基哌嗪衍生物均对 ECV304细胞的生长有明显的抑制作用， 并 与化合物浓度显示良好的线性关系 （见图 1 )。 实施例 13  The results showed that the phenylpiperazine derivatives significantly inhibited the growth of ECV304 cells and showed a good linear relationship with the concentration of the compounds (see Figure 1). Example 13
苯基喊嗪衍生物对于肝癌 HepG-2细胞的抑制活性  Inhibitory activity of phenylpyrazine derivatives on hepatocellular carcinoma HepG-2 cells
实验样品： 分别选取苯基哌嗪化合物 5-氧 -3-苯基 -5-(4-甲苯基哌嗪)戊酸 (JA3031 , 见实施例 3， 编号为 1 ); 3-(4-甲氧苯基) -5-(4-(4-甲氧苯基)哌嗪 -5-氧 -戊酸 （JA3024, 见实施例 6， 编号为 2); 3-(3, 5-二甲氧基苯基 )-5-氧 -5-(4-对甲 苯基哌嗪)戊酸 （JA3040, 见实施例 5， 编号为 3 ); 3-(4-(4-氯苯基)哌嗪） -3-(4- 甲氧基苯基) -5-氧 -戊酸 （JA3036, 见实施例 9， 编号为 4); 3-(3,5-二甲氧基苯 基) -5-(4-(4-乙氧甲酰)苯基)哌嗪） -5-氧 -3-戊酸（JA3023 ,见实施例 8，编号为 5); 3-(3,5-二甲氧基苯基 )-5-(4-苄基哌嗪） -5-氧 -3-戊酸（JA3018, 见实施例 10， 编号 为 6); 4-氧 -4-(4-对甲苯基哌嗪)丁酸（JA3008, 见实施例 4， 编号为 7); 5-(4-(4- 乙氧甲酰苯基)哌嗪) -3-(4-甲氧基苯基) -5-氧 -3-戊酸 （JA3031 , 见实施例 7， 编号 为 8); 4-(4-甲苯磺酰基哌嗪)苯甲酸乙酯 （JA3104, 见实施例 11， 编号为 9) 以 5-Fu作阳性对照。 样品液配置： 分别称取化合物 5mg， 紫外法除菌 20min后将其溶于 0.4ml的 DMSO(Sigma)中， 滴加到装有 9ml的 RPMI- 1640细胞培养液中， 超声溶解， 再 补加 0.6ml的 RPMI-1640细胞培养液质总体积为 10ml。 Experimental sample: phenyl piperazine compound 5-oxo-3-phenyl-5-(4-methylphenylpiperazine) valeric acid (JA3031, see Example 3, No. 1); 3-(4-A Oxyphenyl) -5-(4-(4-methoxyphenyl)piperazine-5-oxo-valeric acid (JA3024, see Example 6, number 2); 3-(3, 5-dimethoxy Phenyl)-5-oxo-5-(4-p-tolylpiperazine) valeric acid (JA3040, see Example 5, No. 3); 3-(4-(4-chlorophenyl)piperazine) -3-(4-methoxyphenyl)-5-oxo-pentanoic acid (JA3036, see Example 9, number 4); 3-(3,5-dimethoxyphenyl)-5-( 4-(4-ethoxycarbonyl)phenyl)piperazine)-5-oxo-3-pentanoic acid (JA3023, see Example 8, number 5); 3-(3,5-dimethoxybenzene 5-(4-benzylpiperazine)-5-oxo-3-pentanoic acid (JA3018, see Example 10, number 6); 4-oxo-4-(4-p-tolylpiperazine) Butyric acid (JA3008, see Example 4, number 7); 5-(4-(4-ethoxyformylphenyl)piperazine)-3-(4-methoxyphenyl)-5-oxo- 3-valeric acid (JA3031, see Example 7, number 8); 4-(4-toluenesulfonylpiperazine)benzoic acid ethyl ester (JA3104, see Example 11, number 9) positive with 5-Fu Control. Sample solution configuration: 5mg of compound was weighed separately, and sterilized by UV method for 20min, dissolved in 0.4ml of DMSO (Sigma), added dropwise to 9ml of RPMI-1640 cell culture medium, sonicated, and then added. The total volume of 0.6 ml of RPMI-1640 cell culture medium was 10 ml.
细胞的培养 将 HepG-2冻存管从低温冰箱中取出， 放入 37°C水浴锅中， 10分钟后拿进细胞房中放在超净操作台上， 将冻存管中的培养液转移至离心管 中， 离心 (1000r/min)5 分钟， 吸去上层液体， 加入 5ml 已配好的培养基 （50ml 的培养基中含有）， 用吸管将细胞吹散， 而后将接种到 50ml的培养瓶中， 放入 二氧化碳培养箱中 （37°C， 5%C02) 进行培养， 隔天细胞贴壁， 更换培养基继 续培养。 Cell culture The HepG-2 cryotube was taken out from the low temperature freezer and placed in a 37 ° C water bath. After 10 minutes, it was taken into the cell room and placed on a clean bench to transfer the culture solution in the cryotube. Into the centrifuge tube, centrifuge (1000r / min) for 5 minutes, aspirate the upper layer of liquid, add 5ml of the prepared medium (containing 50ml of the medium), blow the cells with a pipette, and then inoculate 50ml of culture. The flask was placed in a carbon dioxide incubator (37 ° C, 5% CO 2 ) for culture, and the cells were adhered to the next day, and the medium was changed to continue the culture.
种板： 将培养瓶放在超净台上， 将培养基倒掉， 移取 1ml 的胰酶加入培养 瓶中，平放 30s后，将胰酶倒掉，而后将培养瓶放入二氧化碳培养箱中进行孵化， 3min后将培养瓶放入显微镜下观察， 可见细胞都已经消化漂浮； 向培养瓶中加 入 5ml的培养基， 移取 2ml放入 50ml的离心管中加入培养基稀释成 25ml， 点 板用； 向培养瓶中加入 2ml的培养基， 放入培养箱中继续培养， 待进行第二次 测定。 点板时， 板子的***一圈每孔加入 ΙΟΟμηιΙ的 PBS; 每块板子设置五个调 零孔，加入 ΙΟΟμηιΙ的 RPMI-1640细胞培养液；其它每孔加入 ΙΟΟμηιΙ的细胞液， 放入培养箱中培养 24h至细胞贴壁。  Seed plate: Place the culture bottle on the clean bench, pour the culture medium, remove 1 ml of trypsin into the culture flask, lay it flat for 30 s, then pour the trypsin, and then place the culture flask into the carbon dioxide incubator. Incubate in the medium. After 3 minutes, the culture flask is placed under the microscope to observe that the cells have been digested and floated. Add 5 ml of the medium to the flask, transfer 2 ml into a 50 ml centrifuge tube, add the medium and dilute to 25 ml. For the plate; 2 ml of the medium was added to the flask, and the culture was carried out in the incubator for a second measurement. When the plate is placed, PBS of ΙΟΟμηιΙ is added to each hole of the plate; each plate is provided with five zero-adjusting holes, and RPMI-1640 cell culture solution of ΙΟΟμηιΙ is added; the other cell solution of ΙΟΟμηιΙ is added to each well and placed in the incubator. Culture for 24 h until the cells adhered.
加样： 6个目标化合物和 5-Fu分别加 6个浓度，加入药物原液分别为 100μηι1、 50μηιΚ 25μηι1、 15μηι1、 ΙΟμηιΙ和 5μηι1， 而后每孔加入相应量的 RPMI-1640细 胞培养液， 使每孔都稀释至 ΙΟΟμη Ι, 所稀释得到的样品浓度分别为 0.5mg/ml、 0.25mg/ml、 0.125 mg/mK 0.075 mg/ml、 0.05 mg/ml以及 0.025 mg/ml， 每个浓度 设置 5个复空； 设置 10个孔作为空白对照。 放入 37°C培养箱中培育 48h。  Adding 6 target compounds and 5-Fu to add 6 concentrations, add the drug stock solution to 100μηι1, 50μηιΚ 25μηι1, 15μηι1, ΙΟμηιΙ and 5μηι1, respectively, and then add the corresponding amount of RPMI-1640 cell culture solution per well to make each well. Dilute to ΙΟΟμη Ι, the concentration of the sample obtained by dilution is 0.5mg/ml, 0.25mg/ml, 0.125 mg/mK 0.075 mg/ml, 0.05 mg/ml and 0.025 mg/ml, and each concentration is set to 5 complexes. Empty; set 10 holes as a blank control. Incubate in a 37 ° C incubator for 48 h.
显色及测试： 每孔中加入 20μηι1的 MTT溶液（5mg/mL的 PBS液， 滤过除 菌， 4°C避光保存）， 培养箱中继续培育 4h。 将板子从培养箱中拿出， 放入低温 平板离心仪中离心 2000/5min， 取出， 用注射器吸出培养液， 每孔加入 ΙΟΟμηιΙ 的分析纯的 DMSO, 放入摇床中 150r， 摇 lOmin中， 而后在酶标仪中测定 OD 值。  Color development and test: 20 μηι 1 of MTT solution (5 mg/mL PBS solution, filtered and stored at 4 ° C in the dark) was added to each well, and incubation was continued for 4 hours in the incubator. The plate was taken out from the incubator, placed in a low-temperature plate centrifuge for centrifugation for 2000/5 min, taken out, and the culture solution was aspirated with a syringe. Analytically pure DMSO of ΙΟΟμηιΙ was added to each well, placed in a shaker at 150 r, and shaken for 10 min. The OD value was then determined in a microplate reader.
结果表明， 在较高浓度条件下， 化合物 JA3031 (编号为 1 )、 JA3024 (编号 为 2)和 JA3040 (编号为 3 )及 JA3008 (编号为 7)对于肝癌细胞 hepG-2 的抑 制活性超过阳性对照 5-Fu (见图 2和图 3)( 实施例 14 The results showed that at higher concentrations, compounds JA3031 (numbered 1), JA3024 (numbered 2) and JA3040 (numbered 3) and JA3008 (numbered 7) inhibited hepatocellular carcinoma cell line hepG-2 The activity exceeded the positive control 5-Fu (see Figures 2 and 3) ( Example 14
苯基哌嗪衍生物对于小鼠 S-180肿瘤生长的抑制作用  Inhibition of phenyl piperazine derivatives on tumor growth of mouse S-180
实验样品： 分别选取细胞实验效果较好的三个苯基哌嗪化合物： 5-氧 -3-苯 基 -5-(4-甲苯基哌嗪)戊酸（JA3031, 见实施例 3); 3-(4-甲氧苯基) -5-(4-(4-甲氧苯 基)哌嗪 -5-氧 -戊酸（JA3024, 见实施例 6); 3-(4-(4-氯苯基)哌嗪） -3-(4-甲氧基苯基) -5- 氧 -戊酸（JA3036, 见实施例 9) 为实验样品， 以 5-Fu作阳性对照。  Experimental samples: Three phenyl piperazine compounds with good cell experiments were selected: 5-oxo-3-phenyl-5-(4-methylphenylpiperazine) valeric acid (JA3031, see Example 3); -(4-methoxyphenyl)-5-(4-(4-methoxyphenyl)piperazine-5-oxo-pentanoic acid (JA3024, see Example 6); 3-(4-(4-chloro) Phenyl)piperazine)-3-(4-methoxyphenyl)-5-oxo-pentanoic acid (JA3036, see Example 9) was an experimental sample with 5-Fu as a positive control.
样品液配置， 取约 l.Og化合物样品， 于 lOOmL容量瓶中加入 5mL二甲基 亚砜 （DMSO) 溶解， 蒸馏水稀释， 稀氨水调节 pH为中性， 用孔径为 0.22μηι 滤膜滤过除菌备用。  For the sample solution configuration, take about 1.0 mg of the compound sample, add 5 mL of dimethyl sulfoxide (DMSO) to a 100 mL volumetric flask, dilute with distilled water, adjust the pH to neutral with dilute ammonia water, and filter through a 0.22 μηι filter. The bacteria are ready for use.
每组健康昆明小鼠 10只， 体重 18-20g。 以肉瘤 -180 (S-180) 接种， 第二 天随机按表 1 所述进行分组， 按照设定的剂量， 每天腹腔注射法给药一次， 连 续给药 5天， 以 5-Fu作阳性对照。停药 24小时后， 称体重和肿瘤重量， 肿瘤抑 制率采用公式：  10 healthy Kunming mice in each group, weighing 18-20g. Inoculated with sarcoma-180 (S-180), the rats were randomly grouped as described in Table 1 on the next day. According to the set dose, they were administered once a day by intraperitoneal injection for 5 consecutive days. 5-Fu was used as a positive control. . After 24 hours of withdrawal, weighing and tumor weight, the tumor inhibition rate is based on the formula:
抑 -对照组平均瘤重 合药组平均瘤重 ::100 Inhibition-control group average tumor weight group average tumor weight : 100
对照组平均瘤重 具体实验结果见表 1。 表 1: 部分化合物及联合用药对小鼠 S-180肿瘤生长的抑制作用  The average tumor weight of the control group is shown in Table 1. Table 1: Inhibition of tumor growth by mouse S-180 by some compounds and combination
化合物 药物浓度 平均重量  Compound drug concentration average weight
体重差比 * P 编号 ( mg/ml ) (g) g) (%)  Weight difference ratio * P number ( mg/ml ) (g) g) (%)
JA3031 3 25.4±1.9 -0.023 0.7714±0.26 37.62 (0.01 JA3031 3 25.4±1.9 -0.023 0.7714±0.26 37.62 (0.01
JA3031 6 25.4±1.6 -0.023 0.6945 ±0.23 43.83 (0.01JA3031 6 25.4±1.6 -0.023 0.6945 ±0.23 43.83 (0.01
JA3031 9 21.6±2.4 -0.169 0.6445 ±0.25 47.88 (0.01JA3031 9 21.6±2.4 -0.169 0.6445 ±0.25 47.88 (0.01
JA3024 3 26.6±3.1 0.023 0.9568 ±0.42 22.62 (0.03JA3024 3 26.6±3.1 0.023 0.9568 ±0.42 22.62 (0.03
JA3024 6 27.1±1.2 0.042 0.8879±0.23 28.19 (0.01JA3024 6 27.1±1.2 0.042 0.8879±0.23 28.19 (0.01
JA3024 9 24.5 ±2.5 -0.058 0.6965±0.31 43.68 (0.01JA3024 9 24.5 ±2.5 -0.058 0.6965±0.31 43.68 (0.01
JA3036 3 26.2±1.8 0.008 1.1653±0.28 5.757 (0.05JA3036 3 26.2±1.8 0.008 1.1653±0.28 5.757 (0.05
JA3036 6 23.1±1.3 -0.112 0.6786 ±0.27 45.12 (0.01 阳性对照 5-fu 18.3±4.5 -0.296 0.4414±0.25 64.31 JA3036 6 23.1±1.3 -0.112 0.6786 ±0.27 45.12 (0.01 positive control 5-fu 18.3±4.5 -0.296 0.4414±0.25 64.31
联合用药组 19.3±3.5 -0.258 0.2626±0.11 78.76 (0.01 空白对照组 26.0±3.1 1.2365±0.55  Combined drug group 19.3±3.5 -0.258 0.2626±0.11 78.76 (0.01 blank control group 26.0±3.1 1.2365±0.55
*体重差比为 （实验组小鼠平均重量-对照组小鼠平均重」 1) /对照组小鼠平均重量; **阳性对 照组 5-Fu的浓度为 9.2μ  *The difference in body weight ratio was (average weight of mice in the experimental group - average weight of the control mice) 1) / average weight of the control group mice; ** positive control group 5-Fu concentration was 9.2 μ
共用。 将表 1的结果绘制成图 4， 由图 4可以看出， 苯基哌嗪衍生物对小鼠 S-180 肿瘤的生长具有一定的抑制作用， 在 18μ Μ以上对于 S-180肿瘤生长的抑制作 用超过 40%， 但仍低于 5-Fu。 这可能与它作用的受体的性质有关， 本品不属于 细胞毒性药物。 将苯基哌嗪衍生物与 5-Fu联用 （联合用药组） 其抑制活性明显 超过 5-Fu (达到 78.76%), 表明苯基哌嗪衍生物对细胞毒性的 5-Fu具有增效作 用。 另一方面， 与 5-Fu相比， 苯基哌嗪衍生物的毒副作用明显降低， 化合物实 验组的小鼠的平均重量与空白对照相似（体重减小不超过 18%), 而阳性对照组 (5-Fu) 小鼠体重下降接近 30%。 上述实施例为本发明较佳的实施方式， 但本发明的实施方式并不受上述实 施例的限制， 其他的任何未背离本发明的精神实质与原理下所作的改变、 修饰、 替代、 组合、 简化， 均应为等效的置换方式， 都包含在本发明的保护范围之内。 Share. The results in Table 1 are plotted as Figure 4. It can be seen from Figure 4 that the phenylpiperazine derivative has a certain inhibitory effect on the growth of mouse S-180 tumor, and the inhibition of S-180 tumor growth above 18 μΜ More than 40%, but still below 5-Fu. This may be related to the nature of the receptor it acts on. This product is not a cytotoxic drug. The inhibitory activity of the phenylpiperazine derivative in combination with 5-Fu (combination group) was significantly higher than that of 5-Fu (up to 78.76%), indicating that the phenylpiperazine derivative has a synergistic effect on cytotoxic 5-Fu. . On the other hand, the toxic side effects of the phenylpiperazine derivative were significantly lower than those of 5-Fu, and the average weight of the mice in the compound experimental group was similar to that of the blank control (weight loss did not exceed 18%), while the positive control group (5-Fu) The mice lost weight by nearly 30%. The above embodiments are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and modifications may be made without departing from the spirit and scope of the invention. Simplifications, which are equivalent replacement means, are included in the scope of the present invention.

Claims

权利要求书 Claim
如式 I所示的结构:
Figure imgf000021_0001
Structure as shown in Formula I:
Figure imgf000021_0001
I  I
式中 -R、 -OR、 -COOR、 卤素或 -CN基团， 其中 R代表垸基， 在苯环上的取代位置分别在哌嗪基的对位、 邻位或间位； m为 0〜2的自然数； R2为取代或未取代的羧酸基团或磺酸基团。 Wherein -R, -OR, -COOR, halogen or -CN group, wherein R represents a thiol group, the substitution position on the phenyl ring is in the para, ortho or meta position of the piperazinyl group; m is 0~ a natural number of 2 ; R 2 is a substituted or unsubstituted carboxylic acid group or a sulfonic acid group.
2、根据权利要求 1所述的苯基哌嗪类衍生物及其药学上可接受的盐，其中， 所述 R为 C1〜C11链烃基或带有 3〜7元环的环烃基； 所述卤素为 -F、 -C1或 -Br。  The phenylpiperazine derivative according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R is a C1 to C11 chain hydrocarbon group or a cyclic hydrocarbon group having a 3 to 7 membered ring; Halogen is -F, -C1 or -Br.
3、根据权利要求 1所述的苯基哌嗪类衍生物及其药学上可接受的盐，其中， 所述 R2为： The phenylpiperazine derivative according to claim 1 or a pharmaceutically acceptable salt thereof, wherein the R 2 is:
Figure imgf000021_0002
Figure imgf000021_0002
R3为 -H、 -R、 -OH、 -OR、 -COOH、 -COOR、 卤素或 -CN基团， 其中 R为 垸基； n为 0〜5的自然数。 R 3 is -H, -R, -OH, -OR, -COOH, -COOR, halogen or -CN group, wherein R is a fluorenyl group; n is a natural number from 0 to 5.
4、根据权利要求 1〜3任一项所述的苯基哌嗪类衍生物及其药学上可接受的 盐， 其中所述 m=0， n=l o  The phenylpiperazine derivative according to any one of claims 1 to 3, wherein the m = 0, n = 1 o, and a pharmaceutically acceptable salt thereof
5、 根据权利要求 1所述的一种苯基哌嗪类衍生物及其药学上可接受的盐， 其中所述药学上可接受的盐为钠盐或铵盐。  The phenylpiperazine derivative according to Claim 1 or a pharmaceutically acceptable salt thereof, wherein the pharmaceutically acceptable salt is a sodium salt or an ammonium salt.
6、 一种药物组合物， 其包含权利要求 1所述的苯基哌嗪类衍生物和其药学 上可接受的载体。  A pharmaceutical composition comprising the phenylpiperazine derivative of claim 1 and a pharmaceutically acceptable carrier thereof.
7、根据权利要求 1所述的苯基哌嗪类衍生物及其药学上可接受的盐在制备 抑制肿瘤血管生成和肿瘤细胞转移方面药物中的用途。 The use of the phenylpiperazine derivative according to claim 1 or a pharmaceutically acceptable salt thereof for the preparation of a medicament for inhibiting tumor angiogenesis and tumor cell metastasis.
8、根据权利要求 1所述的苯基哌嗪类衍生物及其药学上可接受的盐在制备 治疗肝癌、 乳腺癌、 卵巢癌、 胃癌、 结肠癌、 肺癌或黑色素瘤肿瘤药物中的用 途。 The use of a phenylpiperazine derivative according to claim 1 or a pharmaceutically acceptable salt thereof for the preparation of a medicament for treating liver cancer, breast cancer, ovarian cancer, gastric cancer, colon cancer, lung cancer or melanoma.
9、根据权利要求 1所述的苯基哌嗪类衍生物及其药学上可接受的盐在制备 作为肿瘤化疗药物和手术治疗中的辅助药物中的用途。  The use of a phenylpiperazine derivative according to claim 1 or a pharmaceutically acceptable salt thereof for the preparation of a medicament for use as a tumor chemotherapeutic drug and for surgical treatment.
10、 根据权利要求 7〜9 中任一项所述的用途， 其中所述药物制成片剂、 胶 囊剂、 颗粒剂、 细粒剂、 粉剂、 丸剂、 贴剂、 口服液或注射剂。  The use according to any one of claims 7 to 9, wherein the drug is formulated into a tablet, a capsule, a granule, a fine granule, a powder, a pill, a patch, an oral solution or an injection.
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Families Citing this family (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102260225B (en) * 2011-06-02 2013-08-21 蒋杰 Phenylpiperazine derivatives for inhibiting tumor metastasis and tumor angiogenesis
CN103965159B (en) * 2014-05-17 2017-04-12 广州医科大学 Ether aryl piperazine derivatives and salt thereof as well as preparation method and application of ether aryl piperazine derivatives
CN105061352A (en) * 2015-07-29 2015-11-18 广州市广金投资管理有限公司 Aryl piperazine derivatives (III), salt thereof, preparation method, and application
CN107879997B (en) * 2017-12-25 2021-01-26 三峡大学 Small molecule inhibitor SLD1338 and application thereof in pharmacy
CN108530391B (en) * 2018-05-17 2022-05-17 袁牧 Amide aryl piperazine derivative and preparation method and application thereof

Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4616086A (en) * 1981-10-08 1986-10-07 Boehringer Mannheim Gmbh Piperazine-substituted aryl and aralkyl carboxylic acids useful for treating infirmaties caused by excess lipids or thrombocyte
EP1219614A1 (en) * 2000-12-22 2002-07-03 The Jordanian Pharmaceutical Manufacturing and Medical Equipment Co.Ltd. Benzene sulfonamides as PDE-V inhibitors for the use against erectile dysfunction
WO2003024946A2 (en) * 2001-09-14 2003-03-27 Merck Patent Gmbh Oxamate derivatives containing a variously substituted nitrogen heterocycle
WO2003082288A1 (en) * 2002-04-03 2003-10-09 Topotarget Uk Limited Carbamic acid compounds comprising a piperazine linkage as hdac inhibitors
CN1642931A (en) * 2002-03-13 2005-07-20 詹森药业有限公司 Sulfonylamino-derivatives as novel inhibitors of histone deacetylase
KR20080094475A (en) * 2007-04-20 2008-10-23 한국화학연구원 Compounds having serotonin receptor affinity
CN101541331A (en) * 2006-08-24 2009-09-23 沃克哈特研究中心 Novel macrolides and ketolides having antimicrobial activity
CN102260225A (en) * 2011-06-02 2011-11-30 广州翰鼎医药科技有限公司 Phenylpiperazine derivatives for inhibiting tumor metastasis and tumor angiogenesis

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US2922788A (en) * 1958-05-26 1960-01-26 Parke Davis & Co Phenylpiperazine compounds and process for producing the same
US5518891A (en) * 1993-03-25 1996-05-21 Actimed Laboratories, Inc. Dye forming composition and detection of hydrogen peroxide therewith
CN102459209A (en) * 2009-04-16 2012-05-16 武田药品工业株式会社 Derivatives of n-acyl-n'-phenylpiperazine useful (inter alia) for the prophylaxis or treatment of diabetes
WO2010121212A2 (en) * 2009-04-17 2010-10-21 H. Lee Moffit Cancer Center And Research Institute, Inc. Indoline scaffold shp-2 inhibitors and method of treating cancer

Patent Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4616086A (en) * 1981-10-08 1986-10-07 Boehringer Mannheim Gmbh Piperazine-substituted aryl and aralkyl carboxylic acids useful for treating infirmaties caused by excess lipids or thrombocyte
EP1219614A1 (en) * 2000-12-22 2002-07-03 The Jordanian Pharmaceutical Manufacturing and Medical Equipment Co.Ltd. Benzene sulfonamides as PDE-V inhibitors for the use against erectile dysfunction
WO2003024946A2 (en) * 2001-09-14 2003-03-27 Merck Patent Gmbh Oxamate derivatives containing a variously substituted nitrogen heterocycle
CN1642931A (en) * 2002-03-13 2005-07-20 詹森药业有限公司 Sulfonylamino-derivatives as novel inhibitors of histone deacetylase
WO2003082288A1 (en) * 2002-04-03 2003-10-09 Topotarget Uk Limited Carbamic acid compounds comprising a piperazine linkage as hdac inhibitors
CN101541331A (en) * 2006-08-24 2009-09-23 沃克哈特研究中心 Novel macrolides and ketolides having antimicrobial activity
KR20080094475A (en) * 2007-04-20 2008-10-23 한국화학연구원 Compounds having serotonin receptor affinity
CN102260225A (en) * 2011-06-02 2011-11-30 广州翰鼎医药科技有限公司 Phenylpiperazine derivatives for inhibiting tumor metastasis and tumor angiogenesis

Non-Patent Citations (5)

* Cited by examiner, † Cited by third party
Title
CHENG GANGYING ET AL.: "Design and antitumor and activity of new integrin alpha,beta, receptor antagonists", PROCEEDINGS OF 2008 ACADEMIC ANNUAL MEETING OF CHINESE PHARMACEUTICAL ASSOCIATION & THE 8TH CONGRESS OF CHINESE PHARMACIST WEEK, October 2008 (2008-10-01), pages 1565 - 1571 *
CHENG; GANGYING: "Design, Synthesis and Activity Study of Arylpiperazines as Integrin avp3 Antagonists", MEDICINE AND HYGIENE, CHINA MASTER'S THESES FULL-TEXT DATABASE, 15 February 2012 (2012-02-15), pages E079 - 37 *
CHOI, S.R. ET AL.: "Generation of Oxamic Acid Libraries: Antimalarials and Inhibitors of Plasmodiumfalciparum Lactate Dehydrogenase", J. COMB. CHEM., vol. 9, no. 2, 31 March 2007 (2007-03-31), pages 292 - 300 *
KAMAL, A. ET AL.: "Base-free monosulfonylation of amines using tosyl or mesyl chloride in water", TETRAHEDRON LETTERS, vol. 49, no. 2, 7 January 2008 (2008-01-07), pages 348 - 353 *
NI, GUANGHUI ET AL.: "Advances in the study of the integrin a(33 antagonists", ACTA PHARMACEUTICASINICA, vol. 41, no. 7, 12 July 2006 (2006-07-12), pages 577 - 582 *

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