| 题名 | S-DABO 类衍生物及AZT-氟喹喏酮类偶联物体外抗HIV 活性及机制研究 |
| 作者 | 龙晶 |
| 学位类别 | 硕士 |
| 答辩日期 | 2008-06 |
| 授予单位 | 中国科学院研究生院 |
| 授予地点 | 北京 |
| 导师 | 郑永唐 |
| 关键词 | HIV 抗HIV-1 抗菌 化学合成类药物 非核苷类抑制剂 S-DABO类化合物 偶联物 氟喹诺酮 金黄色葡萄球菌 |
| 其他题名 | The Anti-HIV Effects and Mechanism of S-DABO Drivatives and AZT-Fluoroquinolones Conjugates |
| 中文摘要 | 本论文由2个相对独立的部分组成: S-DABO类衍生物体外抗HIV活性及其机制研究和AZT－氟喹喏酮类偶联物体外抗HIV-1活性及其机制研究。 HIV逆转录酶抑制剂一直是抗HIV药物研发的热点。该类抑制剂靶定在病毒复制周期早期，为HAART疗法提供了很多新的药物组合。目前FDA批准上市的逆转录酶抑制剂虽然有很多，但由于较严重的毒副作用、HIV病毒易变异、耐药性的出现等问题还需要开发更多的新的逆转录酶抑制剂。本论文对23个S-DABO类化合物和8个AZT－氟喹喏酮类偶联物的体外抗HIV活性进行检测，并对其中活性较高的化合物进行靶点和机制研究。 23个S-DABO类化合物采用对C8166细胞的毒性试验，对HIV-1ⅢB诱导的合胞体形成的抑制试验和对HIV-1ⅢB急性感染的MT-4细胞的保护试验进行抗HIV-1活性初步筛选。试验结果发现所有化合物均对多种HIV宿主细胞毒性小，其中22个化合物具有抗HIV-1活性，特别是化合物RZK-4和RZK-5，其对 HIV-1ⅢB诱导的合胞体形成的SI值（Selective index）分别为>16666和>38462；RZK-4和RZK-5对HIV-1ⅢB急性感染的MT-4细胞的保护的SI值分别为2666.67和2150.54，与相应的阳性对照药品NVP（Nevirapine）的SI值相接近。以p24抗原水平为指标，对其中20个化合物的抗HIV-1活性进行确证，发现这20个化合物均能抑制 HIV-1ⅢB p24抗原的产生，其中RZK-4和RZK-5的EC50值分别为5.93和5.74ng/ml，比相应的阳性对照药品NVP（Nevirapine）的EC50值要低（27.3ng/ml）。这些化合物对试验株HIV-1MN、临床分离株HIV-1KM018、非核苷类抑制剂耐药株HIV-1ⅢB A17也有较好的抑制效果。但这23个S-DABO类化合物对HIV-2病毒株均无抑制作用。通过检测化合物对感染与未感染细胞的融合的抑制、对HIV-1逆转录酶和蛋白酶活性的抑制、对慢性感染H9细胞（H9/HIV-1ⅢB）中病毒复制的抑制等试验来探讨化合物的抗HIV-1机制。结果显示：有20个化合物对HIV-1蛋白酶（PR）有抑制作用，其中有17个化合物对HIV-1逆转录酶（RT）有抑制作用；但所有化合物均不能抑制感染与未感染细胞的融合，也不能抑制慢性感染H9细胞中病毒的复制。试验结果表明，这23个S-DABO类化合物主要通过抑制HIV-1逆转录酶来发挥作用，它们是典型的非核苷类RT抑制剂。 8个AZT－氟喹喏酮类偶联物采用对C8166细胞的毒性试验，对HIV-1ⅢB诱导的合胞体形成的抑制试验和对HIV-1ⅢB急性感染的MT-4细胞的保护试验进行抗HIV-1活性初步筛选。试验结果发现其中2个化合物SRLZ和SROZ有较显著的抗HIV-1活性，其对HIV-1ⅢB诱导的合胞体形成抑制的SI值分别为>41667和>105263；对HIV-1ⅢB急性感染的MT-4细胞的保护的SI值分别为30162 和 6368，与AZT（Zidothymidine）的SI值相近似。以p24抗原水平为指标，对其抗HIV-1活性进行确证，发现化合物SRLZ和SROZ均能抑制HIV-1ⅢB p24抗原的产生，其EC50值分别为 0.71和2.1ng/ml,比相应的阳性对照药品AZT的EC50值要低（3.5ng/ml）。化合物SRLZ和SROZ对临床分离株HIV-1KM018也有较好的抑制活性，其EC50值分别为1.4和22ng/ml。通过检测化合物对慢性感染H9细胞（H9/HIV-1ⅢB）中病毒复制的抑制试验来探讨化合物的抗HIV-1机制，结果表明化合物SRLZ和SROZ均不能抑制慢性感染H9细胞中病毒的复制。通过检测化合物对金黄色葡萄球菌的抑制作用来检测其抗菌活性，化合物SRLZ和SROZ对金黄色葡萄球菌有较好的抑制作用，其MIC（Minimum inhibitory concentration）值分别为14.65和7.32μg/ml，与其相应的阳性对照药物的MIC值相类似。试验结果表明：药物—药物偶连这种化学修饰方法并没有改变AZT－氟喹喏酮类偶联物的抗HIV作用靶点，但也没有较大地影响到其体外抗病毒活性和抗微生物活性。 |
| 英文摘要 | The thesis is composed of two correspondingly independent parts: the activity and the mechanism of S-DABO derivatives on HIV-1 in vitro; anti-HIV-1 activities and the mechanism of AZT-fluoroquinolones conjugates. The study on the inhibitors of HIV-1 reverse transcriptase is a hot area in anti-HIV drugs research. Because of the severious side-effect, the frequently changing of the HIV virus and the arising of drug resistance et al., we need more compounds of this kind, although there already have several reverse transcriptase inhibitors permitted by FDA. Our study tested the anti-HIV-activities and mechanisms of 23 S-DABO derivatives and 8 AZT-Fluoroquinolones conjugates. Through the primary screen, we found that all the 23 S-DABO derivatives showed low cytotoxicity on different cell lines, and 22 of them had anti-HIV-1 activities. The RZK-4 and RZK-5 showed anti-HIV-activites with the selective index (SI) of syncytium formation inhibition of >16666 and >38462 respectivly; and the selective index of protection on HIV-1 infected MT-4 cell were 2667 and 2150. The values of SIs were similar to the NVP’s. 20 of these compounds could decrease the production of HIV-1 p24 antigen, of which the 50% effective concentration (EC50) of RZK-4 and RZK-5 were 5.93 and 5.74ng/ml, the values were lower than the NVP’s（27.3ng/ml）. The activites against another 3 strains of HIV-1 were also determined. There were laboratory-derived virus (HIV-1MN), low-passage clinical isolated virus (HIV-1KM018) and drug-resistant virus of NNRTIs(HIV-1ⅢB A17). These S-DABO derivatives also showed very good anti-HIV-1 effect on these three strains. But all the 23 S-DABO derivatives had no inhibition effect on HIV-2 replication. We tried to find the anti-HIV-1 mechanism of the 23 compounds. Through fusion inhibition assay, RT and PR assay, and inhibition on HIV replication in persistant infected H9 cell assay et al., we found that: 20 compounds showed inhibition effect on the HIV-1 proteinase(PR), and 17 of them had effect on the reverse transcriptase (RT). Whereas, all the compounds could not inhibit fusion in coculture and they had no effect on the HIV replication in persistant infected H9 cell. All the results suggest that the S-DABO derivatives are classic NNRTIs. In 8 AZT-Fluoroquinolones conjugates, SRLZ and SROZ significantly inhibited HIV-1 syncytium formation. Their SI were >416667 and >105263 respectivly. They also had the ability to protect MT-4 cells from HIV-1 infection, the SI were 30162 and 6368. The values were similar to the AZT’s. Both of the two compounds could decrease the production of HIV-1 p24 antigen, and the 50% effective concentration (EC50) were 0.71 and 2.1ng/ml, which were lower than the value of the AZT(3.5 ng/ml). SRLZ and SROZ were also found to be active against HIV-1KM018 (EC50=1.4 and 22ng/ml) infection. The mechanism study showed that the SRLZ and SROZ could not inhibit the HIV-1 replication in persistant infected H9 cell. However, the SRLZ and SROZ showed inhibition effect on the St. aureus growth, the MIC（Minimum inhibitory concentration）value were 14.65 and 7.32μg/ml, which were equal to the positive control drugs. All the results show that, this chemical modification method will not change the drug target of the AZT-Fluoroquinolones conjugates, but it will also not much affect the anti-HIV activities and anti-microbial activities of AZT-Fluoroquinolones conjugates. |
| 语种 | 中文 |
| 公开日期 | 2010-10-13 |
| 内容类型 | 学位论文 |
| 源URL | [http://159.226.149.42:8088/handle/152453/6072]  |
| 专题 | 昆明动物研究所_分子免疫药理学 |
| 推荐引用方式 GB/T 7714 | 龙晶. S-DABO 类衍生物及AZT-氟喹喏酮类偶联物体外抗HIV 活性及机制研究[D]. 北京. 中国科学院研究生院. 2008. |
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