WO2014023063A1 - High efficiency telomerase inhibitor and application of same in antitumor drug - Google Patents
High efficiency telomerase inhibitor and application of same in antitumor drug Download PDFInfo
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- WO2014023063A1 WO2014023063A1 PCT/CN2012/083799 CN2012083799W WO2014023063A1 WO 2014023063 A1 WO2014023063 A1 WO 2014023063A1 CN 2012083799 W CN2012083799 W CN 2012083799W WO 2014023063 A1 WO2014023063 A1 WO 2014023063A1
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- platinum complex
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- trinuclear platinum
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- 239000002246 antineoplastic agent Substances 0.000 title claims abstract description 19
- 229940041181 antineoplastic drug Drugs 0.000 title claims abstract description 18
- 229940123582 Telomerase inhibitor Drugs 0.000 title claims description 7
- 239000003277 telomerase inhibitor Substances 0.000 title claims description 7
- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Chemical compound [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 claims abstract description 133
- 229910052697 platinum Inorganic materials 0.000 claims abstract description 64
- 238000002360 preparation method Methods 0.000 claims abstract description 8
- 239000000126 substance Substances 0.000 claims abstract description 5
- 239000003446 ligand Substances 0.000 claims description 31
- 238000006243 chemical reaction Methods 0.000 claims description 25
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 16
- SQGYOTSLMSWVJD-UHFFFAOYSA-N silver(1+) nitrate Chemical compound [Ag+].[O-]N(=O)=O SQGYOTSLMSWVJD-UHFFFAOYSA-N 0.000 claims description 10
- 230000015572 biosynthetic process Effects 0.000 claims description 9
- 238000006298 dechlorination reaction Methods 0.000 claims description 8
- 239000011261 inert gas Substances 0.000 claims description 8
- 229910052757 nitrogen Inorganic materials 0.000 claims description 8
- 238000003786 synthesis reaction Methods 0.000 claims description 8
- 238000000034 method Methods 0.000 claims description 7
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 7
- 239000007787 solid Substances 0.000 claims description 6
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 5
- 229910001961 silver nitrate Inorganic materials 0.000 claims description 5
- VEXZGXHMUGYJMC-UHFFFAOYSA-M Chloride anion Chemical compound [Cl-] VEXZGXHMUGYJMC-UHFFFAOYSA-M 0.000 claims description 4
- NHNBFGGVMKEFGY-UHFFFAOYSA-N Nitrate Chemical compound [O-][N+]([O-])=O NHNBFGGVMKEFGY-UHFFFAOYSA-N 0.000 claims description 4
- 230000000259 anti-tumor effect Effects 0.000 claims description 4
- 238000001308 synthesis method Methods 0.000 claims description 4
- 229910002651 NO3 Inorganic materials 0.000 claims description 3
- 238000005119 centrifugation Methods 0.000 claims description 3
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- 230000002194 synthesizing effect Effects 0.000 claims description 3
- 239000012467 final product Substances 0.000 claims description 2
- 238000002560 therapeutic procedure Methods 0.000 claims 1
- 238000005406 washing Methods 0.000 claims 1
- 108010017842 Telomerase Proteins 0.000 abstract description 21
- 230000000694 effects Effects 0.000 abstract description 21
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- 102000005936 beta-Galactosidase Human genes 0.000 description 4
- 108010005774 beta-Galactosidase Proteins 0.000 description 4
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- 239000002184 metal Substances 0.000 description 4
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- -1 small molecule compounds Chemical class 0.000 description 4
- 102000053602 DNA Human genes 0.000 description 3
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 3
- 150000001875 compounds Chemical class 0.000 description 3
- 238000011161 development Methods 0.000 description 3
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- 102000055501 telomere Human genes 0.000 description 3
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- 206010008342 Cervix carcinoma Diseases 0.000 description 2
- 208000035896 Twin-reversed arterial perfusion sequence Diseases 0.000 description 2
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- 230000008827 biological function Effects 0.000 description 2
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- 206010058467 Lung neoplasm malignant Diseases 0.000 description 1
- 206010027476 Metastases Diseases 0.000 description 1
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- 206010061306 Nasopharyngeal cancer Diseases 0.000 description 1
- 238000012408 PCR amplification Methods 0.000 description 1
- 102000016971 Proto-Oncogene Proteins c-kit Human genes 0.000 description 1
- 108010014608 Proto-Oncogene Proteins c-kit Proteins 0.000 description 1
- 125000003118 aryl group Chemical group 0.000 description 1
- GRHLMSBCOPRFNA-UHFFFAOYSA-M azanide 2-oxidoacetate platinum(4+) Chemical compound N[Pt]1(N)OCC(=O)O1 GRHLMSBCOPRFNA-UHFFFAOYSA-M 0.000 description 1
- 238000004166 bioassay Methods 0.000 description 1
- 210000001185 bone marrow Anatomy 0.000 description 1
- CBMYFVSIIYILRH-UHFFFAOYSA-N c1cnccc1-c1nc(-c2ccncc2)nc(-c2ccncc2)n1 Chemical compound c1cnccc1-c1nc(-c2ccncc2)nc(-c2ccncc2)n1 CBMYFVSIIYILRH-UHFFFAOYSA-N 0.000 description 1
- 230000004611 cancer cell death Effects 0.000 description 1
- 230000009702 cancer cell proliferation Effects 0.000 description 1
- 229960004562 carboplatin Drugs 0.000 description 1
- YAYRGNWWLMLWJE-UHFFFAOYSA-L carboplatin Chemical compound O=C1O[Pt](N)(N)OC(=O)C11CCC1 YAYRGNWWLMLWJE-UHFFFAOYSA-L 0.000 description 1
- 231100000504 carcinogenesis Toxicity 0.000 description 1
- 230000003197 catalytic effect Effects 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 238000002512 chemotherapy Methods 0.000 description 1
- 229940044683 chemotherapy drug Drugs 0.000 description 1
- DQLATGHUWYMOKM-UHFFFAOYSA-L cisplatin Chemical compound N[Pt](N)(Cl)Cl DQLATGHUWYMOKM-UHFFFAOYSA-L 0.000 description 1
- 229960004316 cisplatin Drugs 0.000 description 1
- 229940121657 clinical drug Drugs 0.000 description 1
- 230000034994 death Effects 0.000 description 1
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- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
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- 125000000524 functional group Chemical group 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 230000001506 immunosuppresive effect Effects 0.000 description 1
- 201000007270 liver cancer Diseases 0.000 description 1
- 208000014018 liver neoplasm Diseases 0.000 description 1
- 201000005202 lung cancer Diseases 0.000 description 1
- 208000020816 lung neoplasm Diseases 0.000 description 1
- 239000006166 lysate Substances 0.000 description 1
- 230000005291 magnetic effect Effects 0.000 description 1
- 229910021645 metal ion Inorganic materials 0.000 description 1
- 230000009401 metastasis Effects 0.000 description 1
- 201000011216 nasopharynx carcinoma Diseases 0.000 description 1
- 229950007221 nedaplatin Drugs 0.000 description 1
- 229910000510 noble metal Inorganic materials 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 229960001756 oxaliplatin Drugs 0.000 description 1
- DWAFYCQODLXJNR-BNTLRKBRSA-L oxaliplatin Chemical compound O1C(=O)C(=O)O[Pt]11N[C@@H]2CCCC[C@H]2N1 DWAFYCQODLXJNR-BNTLRKBRSA-L 0.000 description 1
- 230000001590 oxidative effect Effects 0.000 description 1
- 150000003058 platinum compounds Chemical class 0.000 description 1
- HRGDZIGMBDGFTC-UHFFFAOYSA-N platinum(2+) Chemical compound [Pt+2] HRGDZIGMBDGFTC-UHFFFAOYSA-N 0.000 description 1
- 229920002401 polyacrylamide Polymers 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
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- 238000001959 radiotherapy Methods 0.000 description 1
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- 238000004904 shortening Methods 0.000 description 1
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- 230000033863 telomere maintenance Effects 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 231100000925 very toxic Toxicity 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
- C07F15/00—Compounds containing elements of Groups 8, 9, 10 or 18 of the Periodic Table
- C07F15/0006—Compounds containing elements of Groups 8, 9, 10 or 18 of the Periodic Table compounds of the platinum group
- C07F15/0086—Platinum compounds
- C07F15/0093—Platinum compounds without a metal-carbon linkage
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
Definitions
- the invention relates to a trinuclear platinum complex capable of inhibiting telomerase activity efficiently
- ZD028 and its preparation methods and teachings are applications for antitumor drugs. Specifically, it relates to a method for preparing a trinuclear platinum complex (ZD028) capable of efficiently inhibiting telomerase activity and its use as an antitumor drug. Book
- Nucleic acid-targeted anticancer drugs should be the most promising anticancer drugs because they can fundamentally inhibit the production, development and appreciation of cancer cells.
- G-quadruplex DNA is a special secondary structure with important biological functions.
- the design of anticancer drugs targeting it is reasonable and superior, mainly due to the following characteristics of the structure: 1), structure Specificity: Different from the ubiquitous double-stranded DNA, special structure can provide a design platform for selective small molecule compounds; , the specificity of the function: special biological function, small molecule compound through stability, disassembly or By changing its secondary structure, it can achieve the purpose of specifically inhibiting cancer cells; 3) It exists in important gene regions in the body.
- the complex induces the formation of a G-quadruplex structure and effectively stabilizes the structure, thereby inhibiting the activity of telomerase, destroying the telomere maintenance mechanism of cancer cells, accelerating the senescence of cancer cells, thereby leading to the death of cancer cells, and at the same time due to telomeres.
- the enzyme is inactive in normal cells and has high activity only in cancer cells, so this helps to find significant selectivity against cancer cell proliferation with strong inhibition due to metal complexes with many organic small Molecularly incomparable advantages, such as rich and diverse coordination structures and electronic properties, more importantly, they often have some interesting characteristics, such as optical, magnetic, catalytic functions, etc. These are excellent G-four chains for design. Body stabilizers bring great potential and possibilities. There are more and more kinds of metal ions in the metal complexes of the stabilized G-quadruplex which have been reported so far, and the complexes which have been constructed centering on the noble metal platinum to stabilize the G-quadruplex are not many.
- Platinum complexes as antitumor drugs began in the 1960s, and several platinum complexes (cisplatin, carboplatin, oxaliplatin, nedaplatin, and leplatin, etc.) currently used in clinical drugs.
- Etc.: are mainly nucleic acids targeting a double-stranded structure.
- the metal platinum (II) complex constructed by platinum ion coordination has its unique advantages.
- the bridging ligand is a series of nitrogen-containing heterocycles, which makes the complex easier to ⁇ - ⁇ stack with the G-quadruplex, thereby efficiently stabilizing the structure and thereby improving the inhibition of telomerase activity.
- the present invention simplifies the synthesis, reduces the production cost, and has a high G-quadruplex targeting, high end by introducing specific organic functional groups by supramolecular self-assembly of organic hybrid trinuclear platinum complexes. Granzyme activity inhibition ability and anticancer activity.
- the object of the present invention is to design a trinuclear platinum complex (ZD028) which is highly effective in inhibiting telomerase activity, a preparation method thereof and an application as an antitumor drug.
- the anticancer drug provided by the invention is an organic hybrid trinuclear platinum complex which is self-assembled by supramolecular, and has the following structural formula:
- the invention discloses a method for synthesizing a trinuclear platinum complex having excellent telomerase activity inhibiting ability, which utilizes supramolecular self-assembly and a planar square coordination configuration of Pt(II) and a unique geometric configuration of the intermediate bridging ligand. Made. Furthermore, the supramolecular ligand and the bridging ligand are assembled into a trinuclear structure by supramolecular self-assembly, which has high G-quadruplex targeting, high telomerase activity inhibition ability and anticancer activity. Trinuclear platinum complex.
- the trinuclear platinum complex (referred to as ZD028 in this patent) capable of efficiently inhibiting telomerase activity according to the present invention, and further, the synthesis thereof is a lateral arm monodentate platinum ligand in which chloride ions are replaced by nitrate. [Pt(dien)(N0 3 )](N0 3 ) is self-assembled with the corresponding bridging ligand, so that the complex has high telomerase inhibition ability and anticancer activity.
- (1) is a structural formula of a side-arm monodentate platinum ligand bridging ligand [Pt(dien)(N0 3 )](N0 3 ) in which chloride ions are replaced by nitrate, and (2) is a bridged ligand.
- the structural formula of the bulk, (3) is the structural formula of the final product trinuclear platinum complex (ZD028).
- the method for synthesizing a trinuclear platinum complex (ZD028) having high G-quadruplex targeting, high telomerase inhibition ability and anticancer activity includes, for example, Next steps:
- Monodentate platinum ligand dechlorination The monodentate platinum ligand is dissolved in water, silver nitrate is added, and the reaction is protected from light under the protection of an inert gas, and the night is left after centrifugation.
- Trinuclear platinum complex The bridged ligand is added to the night after dechlorination, and is protected from light under the protection of inert gas. After the reaction is completed, it is washed with absolute ethanol and centrifuged to obtain a solid substance, that is, a target. Trinuclear platinum complex.
- inert gas protection and protection from light are all key steps in the reaction.
- the role of inert gas protection is to isolate air or other oxidizing gases. It is to be understood that any means for achieving the above objects may be used as an alternative to the invention, and such substitutions do not depart from the scope of the invention.
- the inert gas is nitrogen;
- the light-protecting reaction condition of the monodentate platinum ligand dechlorination step is: 45 degree reaction for 36 hours; and the light-protecting reaction condition of the trinuclear platinum complex synthesis step is : 95 degrees in the dark for 3 days.
- the trinuclear platinum complex (ZD028) can be synthesized by the following procedure.
- Dechlorination of monodentate platinum ligand [Pt(dien)Cl]Cl_HCl Dissolve monodentate platinum ligand in an appropriate amount of water and add 3 times the molar amount of silver nitrate, and protect it from light at 45 °C for 36 hours under nitrogen protection. , centrifuge with a low temperature centrifuge, discard the precipitate, and keep it clear.
- Trinuclear platinum complex (ZD028): A bridged ligand having a molar ratio of 0.3:1 was added to the night after dechlorination. The whole reaction was protected by nitrogen at a temperature of 90 ° for 3 days. After the reaction was completed, an appropriate amount of absolute ethanol was added, and the solvent was washed three times, and centrifuged to obtain a solid substance, that is, a trinuclear platinum complex (ZD028). .
- the monodentate platinum ligand of the present invention is a chloride ion substituted with a nitrate ion.
- the present inventors have found that the trinuclear platinum complex has an outstanding effect in inhibiting telomerase and can be used as a telomerase inhibitor. Further, the trinuclear platinum complex of the present invention can be used for the preparation of an antitumor drug.
- the complex when the trinuclear platinum complex (ZD028) is used as a telomerase inhibitor to achieve an antitumor effect, the complex is found to be a series of tumor cells with high expression of telomerase [eg, A549 (human lung cancer cell). ), HepG2 (human liver cancer cells), HeLa (human cervical cancer cells), CNE-2 (human nasopharyngeal carcinoma cells:), MCF-7 (human breast cancer cells: etc.) have excellent telomerase inhibition ability
- telomerase eg, A549 (human lung cancer cell).
- HepG2 human liver cancer cells
- HeLa human cervical cancer cells
- CNE-2 human nasopharyngeal carcinoma cells:
- MCF-7 human breast cancer cells: etc.
- the complex of the present invention can only act on the listed tumor cells, and those skilled in the art should be able to understand that the listed tumor cells are not limited as the scope of protection of the present invention.
- the invention synthesizes and characterizes a high G-quadruplex targeting by a supramolecular self-assembly method by using a simple and easy monodentate platinum ligand as a prosthetic ligand and a corresponding bridging ligand.
- High telomerase inhibitory ability and anti-cancer activity of the trinuclear platinum complex ZD028,.
- the compound has excellent telomerase activity inhibition and anticancer activity by fluorescence energy resonance transfer (FRET) assay, telomerase inhibition (TRAP) assay, long-term cell proliferation assay and ⁇ -galactosidase staining assay.
- FRET fluorescence energy resonance transfer
- TRAP telomerase inhibition
- long-term cell proliferation assay and ⁇ -galactosidase staining assay.
- Figure 1 is a schematic view showing the structure of a trinuclear platinum complex (ZD028);
- Figure 2 shows the ability of the trinuclear platinum complex (ZD028) to inhibit telomerase activity;
- Figure 3 shows the effect of trinuclear platinum complex (ZD028) on long-term proliferation of HeLa cells.
- Figure 4 shows the senescence-associated ⁇ -galactosidase staining assay in HeLa cells.
- the concentration of trinuclear platinum complex (ZD028) is 15 ⁇ .
- ⁇ is the control group and B is the medicated group.
- FRET Fluorescence Energy Resonance Transfer
- 10 ⁇ human telomeric DNA (F21T), promoter DNA (c-kit) or double-stranded DNA (dsDNA) was mixed with 10 different concentrations of complex (ZD028) solution.
- the resulting 20 ⁇ sample solution was added to a LightCycler capillary and inserted into a Roche LightCycler 2 type PCR instrument.
- Fixed excitation wavelength is corrected page (Article 91)
- the emission fluorescence intensity at 530 nm was monitored.
- the temperature range is 37 -99 Q C
- the temperature rise interval is 1 Q C
- the sample is balanced after 30 s
- the m value is calculated using Origin.
- the data results are shown in Table 1.
- the experimental results show that the trinuclear platinum compound can specifically and efficiently stabilize human telomere G-quadruplex DNA.
- ⁇ -galactosidase staining experiment The cells after termination of the culture were fixed, stained, and observed under a 40x microscope. The data results are shown in Figure 4. Since ⁇ -galactosidase is an important feature of cell senescence, it can be seen from Fig. 4 that the complex can effectively accelerate the senescence of cells, thereby achieving an anti-tumor effect.
- the above comprehensive bioassay results show that the trinuclear platinum complex (ZD028) inhibits telomerase activity by acting on telomere G-quadruplex DNA, and interferes with the telomere mechanism of cancer cells, thereby accelerating cancer cells. The shortening of telomere length, which in turn accelerates cell decay, ultimately leads to cancer cell death and is very toxic to normal cells, so the nuclear platinum complex (ZD028) can be used as a potential antitumor drug.
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- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- General Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Pharmacology & Pharmacy (AREA)
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- Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
Abstract
The present invention provides a trinuclear platinum complex (ZD028) capable of inhibiting telomerase activity with high efficiency, a preparation method and an application of same as an antitumor drug. The complex (ZD028) of the present invention has a low toxicity on normal cells, thereby reducing side effects on other organs or systems. The complex has a simple and easy preparation method, is low in costs, can be completed in a common chemical laboratory, and has no environmental pollution during the production process.
Description
一种高效端粒酶抑制剂及其在抗肿瘤药物中的应用 技术领域 High-efficiency telomerase inhibitor and its application in antitumor drugs
本发明涉及一种能高效抑制端粒酶活性的三核铂配合物 The invention relates to a trinuclear platinum complex capable of inhibiting telomerase activity efficiently
(ZD028), 及其制备方法和作说为抗肿瘤药物方面的应用。 具体涉及能 高效抑制端粒酶活性的三核铂配合物 (ZD028)的制备方法及其可作为 抗肿瘤药物的应用。 书 (ZD028), and its preparation methods and teachings are applications for antitumor drugs. Specifically, it relates to a method for preparing a trinuclear platinum complex (ZD028) capable of efficiently inhibiting telomerase activity and its use as an antitumor drug. Book
背景技术 Background technique
目前， 恶性肿瘤是危害人类健康和生命的重大疾病， 但抗肿瘤新 药研发是不断更新， 有中药， 也有西药。 长期大量的临床证明， 抗肿 瘤药物均存在毒副作用较大的通病。外科手术适用于某些局部性肿瘤 早期和中期的治疗，但多数病人靠手术治疗是不能防止肿瘤的复发和 远处转移的。放、 化疗虽然有相当高的治愈率， 但是常引起如骨髓抑 制、 免疫低下等毒副反应， 使患者难以坚持治疗。化疗药物在治疗过 程中出现的耐药性， 已成为目前临床治疗中的难题之一。正是由于这 些原因， 我们正要寻找抗肿瘤新药。 At present, malignant tumors are major diseases that endanger human health and life, but the development of new anti-tumor drugs is constantly updated, with Chinese medicine and western medicine. A long-term large number of clinically proven, anti-tumor drugs have a common side of toxic side effects. Surgery is suitable for early and intermediate treatment of certain local tumors, but most patients cannot prevent tumor recurrence and distant metastasis by surgical treatment. Although radiotherapy and chemotherapy have a relatively high cure rate, they often cause toxic side effects such as bone marrow suppression and immunosuppression, making it difficult for patients to adhere to treatment. The drug resistance of chemotherapeutic drugs in the course of treatment has become one of the problems in current clinical treatment. It is for these reasons that we are looking for new anti-tumor drugs.
以核酸为靶点的抗癌药物应该是最具发展前景的抗癌药物，因为 此类药物可以从根本上抑制癌细胞的产生、 发展和增值。 G-四链体 DNA作为有重要生物功能的特殊二级结构， 以其为靶点的抗癌药物 的设计具有合理性与优越性， 主要由于此结构有以下特征： 1)， 结构
的特殊性： 区别于普遍存在的双链 DNA, 特殊的结构才能为有选择 性的小分子化合物提供设计平台； ， 作用的特殊性： 有特殊的生物 学功能， 小分子化合物通过稳定、 拆散或改变它的这种二级结构， 可 以达到特异性抑制癌细胞的目的； 3)， 存在于体内重要基因区域。 随 着分子生物学和结构生物学的发展， G-四链体的结构多样性和生物功 能性得到不断阐明。 因此， 以各种类型的 G-四链体为作用靶点的药 物研宄引起了人们广泛的兴趣和重视。 配合物诱导形成 G-四链体结 构并有效稳定该结构， 进而抑制端粒酶的活性， 破坏癌细胞的端粒维 持机制， 加速癌细胞的衰老， 进而导致癌细胞的死亡， 同时由于端粒 酶在正常细胞中是没有活性的， 只有在癌细胞中才具有高的活性， 因 此这有助于找到显著的选择性对癌细胞增殖具有强的抑制作用的抗 由于金属配合物具有很多有机小分子难以比拟的优点，如丰富多 样的配位结构及电子性能， 更重要的是它们往往具有一些有趣的特 性， 例如光学、 磁学、 催化功能等等， 这些为设计成为优秀的 G-四 链体稳定剂带来很大的潜力与可能性。 目前报道的稳定 G-四链体的 金属配合物中金属离子的种类已越来越多，到现在为止以贵金属铂为 中心构建的配合物来稳定 G-四链体还不是很多。 铂类配合物作为抗 肿瘤药物的研宄开始于二十世纪六十年代， 目前使用于临床药物的几 种铂配合物 (顺铂、 卡铂、 奥沙利铂、 奈达铂以及乐铂等等：)都是以靶 向双链结构的核酸为主。 尽管以 G-四链体为靶的选择性抗癌药物的 研宄也只有 10多年， 但已陆续开始有新的小分子化合物进入临床实
验， 不过这些化合物中还没有金属配合物。 Nucleic acid-targeted anticancer drugs should be the most promising anticancer drugs because they can fundamentally inhibit the production, development and appreciation of cancer cells. G-quadruplex DNA is a special secondary structure with important biological functions. The design of anticancer drugs targeting it is reasonable and superior, mainly due to the following characteristics of the structure: 1), structure Specificity: Different from the ubiquitous double-stranded DNA, special structure can provide a design platform for selective small molecule compounds; , the specificity of the function: special biological function, small molecule compound through stability, disassembly or By changing its secondary structure, it can achieve the purpose of specifically inhibiting cancer cells; 3) It exists in important gene regions in the body. With the development of molecular biology and structural biology, the structural diversity and biological functionality of G-quadruplexes have been continuously elucidated. Therefore, drug research with various types of G-quadruplex as a target has attracted wide interest and attention. The complex induces the formation of a G-quadruplex structure and effectively stabilizes the structure, thereby inhibiting the activity of telomerase, destroying the telomere maintenance mechanism of cancer cells, accelerating the senescence of cancer cells, thereby leading to the death of cancer cells, and at the same time due to telomeres. The enzyme is inactive in normal cells and has high activity only in cancer cells, so this helps to find significant selectivity against cancer cell proliferation with strong inhibition due to metal complexes with many organic small Molecularly incomparable advantages, such as rich and diverse coordination structures and electronic properties, more importantly, they often have some interesting characteristics, such as optical, magnetic, catalytic functions, etc. These are excellent G-four chains for design. Body stabilizers bring great potential and possibilities. There are more and more kinds of metal ions in the metal complexes of the stabilized G-quadruplex which have been reported so far, and the complexes which have been constructed centering on the noble metal platinum to stabilize the G-quadruplex are not many. Platinum complexes as antitumor drugs began in the 1960s, and several platinum complexes (cisplatin, carboplatin, oxaliplatin, nedaplatin, and leplatin, etc.) currently used in clinical drugs. Etc.:) are mainly nucleic acids targeting a double-stranded structure. Although the research on selective anticancer drugs targeting G-quadruplex has only been for more than 10 years, new small molecule compounds have begun to enter clinical practice. However, there are no metal complexes in these compounds.
考虑到 G-四链体的芳香平面结构特点， 比较而言由铂离子配位 构建的金属铂 (II)配合物具有其独特的优越性。 例如本专利所涉及的 三核铂配合物 (ZD028)。 Considering the aromatic planar structure of G-quadruplex, the metal platinum (II) complex constructed by platinum ion coordination has its unique advantages. For example, the trinuclear platinum complex (ZD028) referred to in this patent.
桥联配体是一系列含氮的杂环， 使配合物更容易和 G-四链体发 生 π-π堆积作用， 进而能高效的稳定此结构， 进而提高端粒酶活性的 抑制能力。 The bridging ligand is a series of nitrogen-containing heterocycles, which makes the complex easier to π-π stack with the G-quadruplex, thereby efficiently stabilizing the structure and thereby improving the inhibition of telomerase activity.
本发明通过超分子自组装有机杂化三核铂配合物， 简化其合成， 降低生产成本， 并通过引入特定的有机功能基团使其具有高的 G-四 链体靶向性、 高的端粒酶活性抑制能力以及抗癌活性。 The present invention simplifies the synthesis, reduces the production cost, and has a high G-quadruplex targeting, high end by introducing specific organic functional groups by supramolecular self-assembly of organic hybrid trinuclear platinum complexes. Granzyme activity inhibition ability and anticancer activity.
发明内容 Summary of the invention
本发明目的是设计出一种涉及能高效抑制端粒酶活性的三核铂 配合物 (ZD028), 及其制备方法和作为抗肿瘤药物方面的应用。 The object of the present invention is to design a trinuclear platinum complex (ZD028) which is highly effective in inhibiting telomerase activity, a preparation method thereof and an application as an antitumor drug.
本发明提供的抗癌药物，是利用超分子自组装而成的有机杂化三 核铂配合物， 结构式如下： The anticancer drug provided by the invention is an organic hybrid trinuclear platinum complex which is self-assembled by supramolecular, and has the following structural formula:
本发明具有优异端粒酶活性抑制能力的三核铂配合物的合成方 法， 利用超分子自组装和 Pt(II)的平面正方形的配位构型以及中间桥 联配体独特的几何构型而制得的。进一步说， 是利用超分子自组装将 辅基配体与桥联配体组装成三核结构， 得到具有高的 G-四链体靶向 性、 高的端粒酶活性抑制能力以及抗癌活性的三核铂配合物。 The invention discloses a method for synthesizing a trinuclear platinum complex having excellent telomerase activity inhibiting ability, which utilizes supramolecular self-assembly and a planar square coordination configuration of Pt(II) and a unique geometric configuration of the intermediate bridging ligand. Made. Furthermore, the supramolecular ligand and the bridging ligand are assembled into a trinuclear structure by supramolecular self-assembly, which has high G-quadruplex targeting, high telomerase activity inhibition ability and anticancer activity. Trinuclear platinum complex.
本发明所述的能高效抑制端粒酶活性的三核铂配合物 （在本专 利中命名为 ZD028), 更进一步说， 其合成是将氯离子被硝酸根取代 的侧臂单齿铂配体 [Pt(dien)(N03)](N03)与相应的桥联配体通过自组 装而成， 使得该配合物具有高的端粒酶抑制能力以及抗癌活性。 The trinuclear platinum complex (referred to as ZD028 in this patent) capable of efficiently inhibiting telomerase activity according to the present invention, and further, the synthesis thereof is a lateral arm monodentate platinum ligand in which chloride ions are replaced by nitrate. [Pt(dien)(N0 3 )](N0 3 ) is self-assembled with the corresponding bridging ligand, so that the complex has high telomerase inhibition ability and anticancer activity.
该三核铂配合物 (ZD028)的合成方法， 反应式如下： The synthesis method of the trinuclear platinum complex (ZD028) has the following reaction formula:
反应式中， （1)是氯离子被硝酸根取代的侧臂单齿铂配体桥联配 体 [Pt(dien)(N03)](N03)的结构式，（2)是桥联配体的结构式，（3)是最终 产物三核铂配合物 (ZD028)的结构式。 In the reaction formula, (1) is a structural formula of a side-arm monodentate platinum ligand bridging ligand [Pt(dien)(N0 3 )](N0 3 ) in which chloride ions are replaced by nitrate, and (2) is a bridged ligand. The structural formula of the bulk, (3) is the structural formula of the final product trinuclear platinum complex (ZD028).
具体来说， 本发明提供的具有高的 G-四链体靶向性、高的端粒酶 抑制能力以及抗癌活性的三核铂配合物 (ZD028)的合成方法， 包括如
下步骤： Specifically, the method for synthesizing a trinuclear platinum complex (ZD028) having high G-quadruplex targeting, high telomerase inhibition ability and anticancer activity provided by the present invention includes, for example, Next steps:
单齿铂配体脱氯：将所述的单齿铂配体溶于水中，再加入硝酸银， 在惰性气体保护下避光反应， 离心后保留清夜。 Monodentate platinum ligand dechlorination: The monodentate platinum ligand is dissolved in water, silver nitrate is added, and the reaction is protected from light under the protection of an inert gas, and the night is left after centrifugation.
三核铂配合物合成： 往上述脱氯后的清夜中加入所述的桥联配 体， 在惰性气体保护下避光反应， 反应完毕后， 加入无水乙醇洗涤， 离心得到固体物质， 即目标三核铂配合物。 Synthesis of trinuclear platinum complex: The bridged ligand is added to the night after dechlorination, and is protected from light under the protection of inert gas. After the reaction is completed, it is washed with absolute ethanol and centrifuged to obtain a solid substance, that is, a target. Trinuclear platinum complex.
上述的惰性气体保护及避光均为反应的关键步骤。其中惰性气体 保护的作用，是在于隔绝空气或其他具有氧化性的气体。应当理解为， 任何一种可以实现上述目的的手段， 都可以作为替换手段用于本发 明， 并且这种替换并不脱离本发明的保护范围。 The above inert gas protection and protection from light are all key steps in the reaction. The role of inert gas protection is to isolate air or other oxidizing gases. It is to be understood that any means for achieving the above objects may be used as an alternative to the invention, and such substitutions do not depart from the scope of the invention.
作为优选的方案， 所述的惰性气体为氮气； 单齿铂配体脱氯步骤 的避光反应条件为： 45度反应 36小时； 所述的三核铂配合物合成步骤 的避光反应条件为： 95度避光反应 3天。 Preferably, the inert gas is nitrogen; the light-protecting reaction condition of the monodentate platinum ligand dechlorination step is: 45 degree reaction for 36 hours; and the light-protecting reaction condition of the trinuclear platinum complex synthesis step is : 95 degrees in the dark for 3 days.
可以采用以下步骤来合成所述三核铂配合物 (ZD028)。 The trinuclear platinum complex (ZD028) can be synthesized by the following procedure.
单齿铂配体 [Pt(dien)Cl]Cl_HCl脱氯： 将单齿铂配体溶于适量的水 中再加入其 3倍摩尔量的硝酸银， 于 45度在氮气保护下避光反应 36小 时， 用低温离心机离心， 弃去沉淀， 保留清夜。 Dechlorination of monodentate platinum ligand [Pt(dien)Cl]Cl_HCl: Dissolve monodentate platinum ligand in an appropriate amount of water and add 3 times the molar amount of silver nitrate, and protect it from light at 45 °C for 36 hours under nitrogen protection. , centrifuge with a low temperature centrifuge, discard the precipitate, and keep it clear.
三核铂配合物 (ZD028) : 往上述脱氯后的清夜中加入摩尔比为 0.3:1的桥联配体。 整个反应在氮气的保护下， 于 90度避光反应 3天， 反应完毕后， 加入适量的无水乙醇， 再用该溶剂洗涤 3次， 离心得到 固体物质， 即三核铂配合物 (ZD028)。 Trinuclear platinum complex (ZD028): A bridged ligand having a molar ratio of 0.3:1 was added to the night after dechlorination. The whole reaction was protected by nitrogen at a temperature of 90 ° for 3 days. After the reaction was completed, an appropriate amount of absolute ethanol was added, and the solvent was washed three times, and centrifuged to obtain a solid substance, that is, a trinuclear platinum complex (ZD028). .
本发明所述的单齿铂配体为氯离子被硝酸根离子取代的。
本发明发现，所述的三核铂配合物在抑制端粒酶方面具有突出的 效果， 可以作为端粒酶抑制剂。进一步地说， 本发明的三核铂配合物 可以被用于制备抗肿瘤药物。 The monodentate platinum ligand of the present invention is a chloride ion substituted with a nitrate ion. The present inventors have found that the trinuclear platinum complex has an outstanding effect in inhibiting telomerase and can be used as a telomerase inhibitor. Further, the trinuclear platinum complex of the present invention can be used for the preparation of an antitumor drug.
本发明在研宄三核铂配合物 (ZD028)作为端粒酶抑制剂来达到抗 肿瘤效果时， 发现该配合物对一系列端粒酶呈高表达的肿瘤细胞 [例 如: A549(人肺癌细胞)， HepG2(人肝癌细胞)， HeLa(人宫颈癌细胞)， CNE-2(人鼻咽癌细胞：)， MCF-7(人乳腺癌细胞:)等]均具有优异的端粒 酶抑制能力， 但是由于宫颈癌居妇女癌症发病率之首且 HeLa细胞中 端粒酶的活性是非常的高并数量也非常的多，所以以下我们的实施例 主要是以 HeLa细胞为试验对象。 但这并不意味着本发明所述配合物 仅能作用于所列举的肿瘤细胞， 本领域技术人员应当能够理解， 所列 举的肿瘤细胞不能作为本发明保护范围的限制。 In the present invention, when the trinuclear platinum complex (ZD028) is used as a telomerase inhibitor to achieve an antitumor effect, the complex is found to be a series of tumor cells with high expression of telomerase [eg, A549 (human lung cancer cell). ), HepG2 (human liver cancer cells), HeLa (human cervical cancer cells), CNE-2 (human nasopharyngeal carcinoma cells:), MCF-7 (human breast cancer cells: etc.) have excellent telomerase inhibition ability However, because cervical cancer is the leading cause of cancer among women and the activity of telomerase in HeLa cells is very high and the number is very high, the following examples are mainly based on HeLa cells. However, this does not mean that the complex of the present invention can only act on the listed tumor cells, and those skilled in the art should be able to understand that the listed tumor cells are not limited as the scope of protection of the present invention.
本发明具有以下的技术效果： The invention has the following technical effects:
本发明以简单易合的单齿铂配体为辅基配体与相应的桥联配体， 通过超分子自组装的方法合成和表征了一种具有高的 G-四链体靶向 性、 高的端粒酶抑制能力以及抗癌活性的三核铂配合物 (ZD028)。 通 过荧光能量共振转移 (FRET)实验、 端粒酶抑制 (TRAP)实验、 长期细 胞增殖实验和 β-半乳糖苷酶染色实验证明了该化合物具有优异的端 粒酶活性抑制能力以及抗癌活性。 The invention synthesizes and characterizes a high G-quadruplex targeting by a supramolecular self-assembly method by using a simple and easy monodentate platinum ligand as a prosthetic ligand and a corresponding bridging ligand. High telomerase inhibitory ability and anti-cancer activity of the trinuclear platinum complex (ZD028). The compound has excellent telomerase activity inhibition and anticancer activity by fluorescence energy resonance transfer (FRET) assay, telomerase inhibition (TRAP) assay, long-term cell proliferation assay and β-galactosidase staining assay.
附图说明 DRAWINGS
图 1为三核铂配合物 (ZD028)的结构示意图； Figure 1 is a schematic view showing the structure of a trinuclear platinum complex (ZD028);
图 2显示了三核铂配合物 (ZD028)对端粒酶活性的抑制能力；
图 3显示了三核铂配合物 (ZD028)对 HeLa细胞长期增殖的影响 图 4为 HeLa细胞中衰老相关的 β -半乳糖苷酶染色实验， 三核铂配 合物 (ZD028)浓度为 15 μΜ, Α为对照组， B为加药组。 Figure 2 shows the ability of the trinuclear platinum complex (ZD028) to inhibit telomerase activity; Figure 3 shows the effect of trinuclear platinum complex (ZD028) on long-term proliferation of HeLa cells. Figure 4 shows the senescence-associated β-galactosidase staining assay in HeLa cells. The concentration of trinuclear platinum complex (ZD028) is 15 μΜ. Α is the control group and B is the medicated group.
具体实施方式 detailed description
以下结合实施例对本发明进行详细的描述。 The invention will now be described in detail in connection with the embodiments.
实施例一 Embodiment 1
有机杂化三核铂配合物的合成 (ZD028): 将 0.5毫摩尔量的单齿铂 配体 [Pt(dien)Cl]Cl*HCl溶于 6毫升水中， 在氮气保护下于暗处加入 1.5 毫摩尔量的硝酸银， 45度搅拌 36小时， 反应完毕后， 离心弃去沉淀， 保留清夜； 往上述的清液中加入 0.15毫摩尔的桥联配体， 整个反应在 氮气保护下于 90度避光反应 3天， 反应结束后， 往反应液中加入适量 的无水乙醇， 析出浅黄色固体， 离心得浅黄色固体， 产物真空干燥。 产率: 83%。 元素分析(％)， Synthesis of organic hybrid trinuclear platinum complex (ZD028): Dissolve 0.5 mmol of monodentate platinum ligand [Pt(dien)Cl]Cl*HCl in 6 ml of water and add 1.5 in the dark under nitrogen protection. A millimolar amount of silver nitrate was stirred at 45 degrees for 36 hours. After the reaction was completed, the precipitate was discarded by centrifugation and left at night; 0.15 mmol of the bridging ligand was added to the above supernatant, and the whole reaction was 90 degree under nitrogen protection. After the reaction was completed for 3 days, after the reaction was completed, an appropriate amount of absolute ethanol was added to the reaction mixture to precipitate a pale yellow solid, which was evaporated to give a pale yellow solid. Yield: 83%. Elemental analysis(%),
理论值： C30H51N21O18Pt3 6H2O: C， 21.36; H， 3.76; N, 17.43. 实验值： C, 21.20; H， 3.76; N, 17.52. 195Pt NMR (D20, δ/ppm): -1222.20, and Theoretical value: C 30 H 51 N 21 O 18 Pt 3 6H 2 O: C, 21.36; H, 3.76; N, 17.43. Found: C, 21.20; H, 3.76; N, 17.52. 195 Pt NMR (D 2 0, δ/ppm): -1222.20, and
K2PtCl4被用作内标物 (δ=0). K 2 PtCl 4 was used as an internal standard (δ = 0).
该化合物的结构式如图 1所示。 The structural formula of this compound is shown in Figure 1.
实施例二 Embodiment 2
荧光能量共振转移 (FRET)实验： 将 10 μΜ人类端粒 DNA (F21T)、 启动子 DNA (c-kit)或双链 DNA (dsDNA)与 10 不同浓度的配合物 (ZD028)溶液混合。将所得的 20μί样品溶液加入 LightCycler 毛细管中 并插入 Roche LightCycler 2型荧光定量 PCR仪。 固定激发波长为 更正页 （细则第 91条)
470nm， 监测 530nm的发射荧光强度。 温度变化范围为 37 -99 QC,升温 间隔为 1 QC， 平衡 30 s 后采样， m值计算用 Origin拟合。 数据结果见 表 1。实验结果表明该三核铂化合物能特异性的高效的稳定人体端粒 G-四链体 DNA。 Fluorescence Energy Resonance Transfer (FRET) Experiment: 10 μΜ human telomeric DNA (F21T), promoter DNA (c-kit) or double-stranded DNA (dsDNA) was mixed with 10 different concentrations of complex (ZD028) solution. The resulting 20 μί sample solution was added to a LightCycler capillary and inserted into a Roche LightCycler 2 type PCR instrument. Fixed excitation wavelength is corrected page (Article 91) At 470 nm, the emission fluorescence intensity at 530 nm was monitored. The temperature range is 37 -99 Q C, the temperature rise interval is 1 Q C, the sample is balanced after 30 s, and the m value is calculated using Origin. The data results are shown in Table 1. The experimental results show that the trinuclear platinum compound can specifically and efficiently stabilize human telomere G-quadruplex DNA.
核铂配合物 (ZD028)对不同 DNA热稳定能力 (Arm)。
实施例三 The nuclear stabilizing capacity (Ar m ) of the nuclear platinum complex (ZD028) for different DNAs. Embodiment 3
端粒酶抑制 CTRAP)实验:收集 HeLa细胞的裂解液， 加入到含有相 应浓度配合物的 TRAP反应液中， 进行 PCR扩增， 用 0.8%的聚丙烯酰 胺凝胶分析其扩增产物。 数据结果见图 2。 实验结果表明， 该配合物 是一个优异的端粒酶抑制剂 （Td/C5。=30 nM)。 Telomerase inhibition CTRAP): The lysate of HeLa cells was collected, added to the TRAP reaction solution containing the corresponding concentration of the complex, and subjected to PCR amplification, and the amplified product was analyzed with a 0.8% polyacrylamide gel. The data results are shown in Figure 2. The experimental results show that the complex is an excellent telomerase inhibitor ( Td / C 5 = 30 nM).
实施例四 Embodiment 4
长期细胞增殖实验： 将 HeLa细胞接种于 T25组织培养瓶中， 每四 天加入含有相应浓度配合物的培养基或含 0.1%DMSO的培养基。数据 结果见图 3。细胞长期增值实验表明， 该配合物能够降低细胞的生存 率， 进而达到抗肿瘤的效果。 Long-term cell proliferation assay: HeLa cells were seeded in T25 tissue culture flasks, and medium containing the corresponding concentration of complex or medium containing 0.1% DMSO was added every four days. The data results are shown in Figure 3. Long-term cell proliferation experiments show that the complex can reduce the survival rate of the cells and achieve anti-tumor effects.
实施例五 Embodiment 5
β-半乳糖苷酶染色实验： 对培养终止后的细胞进行固定、 染色， 置于 40x显微镜下观察、 照相。 数据结果见图 4。 因为 β-半乳糖苷酶 是细胞衰老的一个重要特征， 通过图 4可知， 该配合物能有效的加速 细胞的衰老， 进而达到抗肿瘤的效果。
以上综合的生测实验结果表明， 三核铂配合物 (ZD028)通过作用 端粒 G-四链体 DNA抑制了端粒酶的活性， 干扰了癌细胞端粒的维 机制， 从而加速了癌细胞端粒长度的縮短， 并进而加速了细胞的衰 ， 最终导致了癌细胞的死亡， 并对正常细胞的毒性非常低， 因此该 核铂配合物 (ZD028)可作为一种潜在的抗肿瘤药物。
Β-galactosidase staining experiment: The cells after termination of the culture were fixed, stained, and observed under a 40x microscope. The data results are shown in Figure 4. Since β-galactosidase is an important feature of cell senescence, it can be seen from Fig. 4 that the complex can effectively accelerate the senescence of cells, thereby achieving an anti-tumor effect. The above comprehensive bioassay results show that the trinuclear platinum complex (ZD028) inhibits telomerase activity by acting on telomere G-quadruplex DNA, and interferes with the telomere mechanism of cancer cells, thereby accelerating cancer cells. The shortening of telomere length, which in turn accelerates cell decay, ultimately leads to cancer cell death and is very toxic to normal cells, so the nuclear platinum complex (ZD028) can be used as a potential antitumor drug.
Claims
1、 一种三核铂配合物， 其特征在于结构式如下: 1. A trinuclear platinum complex characterized by the following structural formula:
2、根据权利要求 1所述的三核铂配合物的制备方法， 其特征在于 反应式如下: 2. The preparation method of the trinuclear platinum complex according to claim 1, characterized in that the reaction formula is as follows:
反应式中，（1)是氯离子被硝酸根取代的单齿铂配体的结构式， (2) 是桥联配体的结构式， （3)是最终产物三核铂配合物的结构式。 In the reaction formula, (1) is the structural formula of a monodentate platinum ligand in which chloride ion is replaced by nitrate, (2) is the structural formula of the bridging ligand, and (3) is the structural formula of the final product trinuclear platinum complex.
3、根据权利要求 2所述的该三核铂配合物的制备方法， 其特征在 于反应步骤如下： 3. The preparation method of the trinuclear platinum complex according to claim 2, characterized in that the reaction steps are as follows:
单齿铂配体脱氯：将所述的单齿铂配体溶于水中，再加入硝酸银，
在惰性气体保护下避光反应， 离心后保留清夜； Dechlorination of single-dentate platinum ligand: Dissolve the single-dentate platinum ligand in water, then add silver nitrate, React in the dark under the protection of inert gas, and keep it clear after centrifugation;
三核铂配合物合成： 往上述脱氯后的清夜中加入所述的桥联配 体， 在惰性气体保护下避光反应， 反应完毕后， 加入无水乙醇洗涤， 离心得到固体物质， 即目标三核铂配合物。 Synthesis of trinuclear platinum complex: Add the bridging ligand to the above-mentioned dechlorinated clear water, and react in the dark under the protection of inert gas. After the reaction is completed, add absolute ethanol for washing, and centrifuge to obtain a solid substance, which is the target. Trinuclear platinum complex.
4、根据权利要求 3所述的三核铂配合物的合成方法， 其特征在于 所述的惰性气体为氮气。 4. The method for synthesizing a trinuclear platinum complex according to claim 3, characterized in that the inert gas is nitrogen.
5、根据权利要求 3所述的三核铂配合物的合成方法， 其特征在于 所述的单齿铂配体脱氯步骤的避光反应条件为： 45度反应 36小时。 5. The synthesis method of trinuclear platinum complex according to claim 3, characterized in that the light-protected reaction conditions of the dechlorination step of the monodentate platinum ligand are: reaction at 45 degrees for 36 hours.
6、根据权利要求 3所述的三核铂配合物的合成方法， 其特征在于 所述的三核铂配合物合成步骤的避光反应条件为： 95度避光反应 3天。 6. The synthesis method of trinuclear platinum complex according to claim 3, characterized in that the light-protected reaction conditions of the trinuclear platinum complex synthesis step are: 95 degrees light-protected reaction for 3 days.
7、根据权利要求 3所述的三核铂配合物的合成方法， 其特征在于 反应步骤如下： 7. The synthesis method of trinuclear platinum complex according to claim 3, characterized in that the reaction steps are as follows:
单齿铂配体脱氯： 将所述单齿铂配体溶于适量的水中再加入 3倍 摩尔量的硝酸银，在氮气保护下避光反应 36小时，用低温离心机离心， 弃去沉淀， 保留清夜； Dechlorination of single-dentate platinum ligand: Dissolve the single-dentate platinum ligand in an appropriate amount of water, add 3 times the molar amount of silver nitrate, react in the dark under nitrogen protection for 36 hours, centrifuge with a low-temperature centrifuge, and discard the precipitate. , keep the clear night;
三核铂配合物合成： 往上述脱氯后的清夜中加入摩尔比为 0.3:1 的所述的桥联配体， 氮气的保护下， 于 95度避光反应 3天， 反应完毕 后， 加入无水乙醇， 离心得到固体物质， 即目标三核铂配合物。 Synthesis of trinuclear platinum complex: Add the bridging ligand with a molar ratio of 0.3:1 to the above-mentioned dechlorinated clear water, and react under the protection of nitrogen for 3 days in the dark at 95 degrees. After the reaction is completed, add Absolute ethanol, and centrifuge to obtain a solid substance, which is the target trinuclear platinum complex.
8、根据权利要求 1所述的三核铂配合物在作为端粒酶抑制剂方面 的应用。 8. Use of the trinuclear platinum complex as a telomerase inhibitor according to claim 1.
9、根据权利要求 1所述的三核铂配合物在制备抗肿瘤药物中的应 用。
9. Use of the trinuclear platinum complex according to claim 1 in the preparation of anti-tumor drugs.
10、 根据权利要求 1所述的三核铂配合物作为高效的端粒酶抑审 L 剂在抗肿瘤治疗联合用药方面的应用
10. Application of the trinuclear platinum complex according to claim 1 as an efficient telomerase inhibitor in combination anti-tumor therapy
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| CN102250150A (en) * | 2011-05-17 | 2011-11-23 | 中山大学 | Organic hydridized tetra-core platinum complex and preparation method thereof as well as application technology field of complex in antitumor medicament preparation |
| CN102408452A (en) * | 2011-12-13 | 2012-04-11 | 中山大学 | Tetrapyridylporphine bridged crossed tetra-palladium complexes, and preparation method and antitumor activity thereof |
| CN102516315A (en) * | 2011-12-12 | 2012-06-27 | 中山大学 | Trinuclear platinum complex possessing Y type structure and its targeting for gastric adenocarcinoma cells |
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| CN102250150A (en) * | 2011-05-17 | 2011-11-23 | 中山大学 | Organic hydridized tetra-core platinum complex and preparation method thereof as well as application technology field of complex in antitumor medicament preparation |
| CN102516315A (en) * | 2011-12-12 | 2012-06-27 | 中山大学 | Trinuclear platinum complex possessing Y type structure and its targeting for gastric adenocarcinoma cells |
| CN102408452A (en) * | 2011-12-13 | 2012-04-11 | 中山大学 | Tetrapyridylporphine bridged crossed tetra-palladium complexes, and preparation method and antitumor activity thereof |
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