Colaborative Research on Anti-HIV Triterpenoids

抗HIV三萜类化合物的合作研究

基本信息

批准号：
09044339
负责人：
KASHIWADA Yoshiki
金额：
$ 1.09万
依托单位：
Niigata College of Pharmacy
依托单位国家：
日本
项目类别：
Grant-in-Aid for international Scientific Research
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1998
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09044339/
关键词：
Anti-HIV Triterpenoids Betulinic acid Oleanolic acid Ursolic acid ベツリン酸

项目摘要

A series of lupane-type triterpenoic acid derivatives were synthesized and evaluated for their inhibitory activity against HIV-1 replication in acutely infected H9 cells, based on the fact that betulinic acid (1) and dihydrobetulinic acid (9) were identified as anti-HIV agents. Among the derivatives, 3-O-(3 ', 3 '-dimethylsuccinyl)-betulinic acid (3) and 3-O-(3 ', 3 '-dimethyl-succinyl)-dihydrobetulinic acid (11) both demonstrated extremely potent inhibitory activity with EC_<50> values of< 0.00035 muM.They exhibited remarkable in vitro therapeutic index (TI.) values of >20,000 and >14,000, respectively. 3-O-(3', 3'-Dimethylglutaryl)-betulinic acid and -dihydrobetulinic acid(12), 3-O-diglycolyl-betulinic acid and -dihydrobetulinic acid and 3-O-glutaryl- betulinic acid were also potent inhibitors of HIV replication with ED_<50> values ranging from 0.01 to 0.0023 muM and T.I.values from 1,017 to 2,344. In addition, compounds 1 1 and 12 are also active against HLV replica-tion in a monocy … More te cell line and n peripheral blood mononuclear cells (PBMCs). Our in vitro assay indicated that these compounds are not inhibitors on HIV- 1 reverse transcriptase, whereas they inhibited syncytia formation completely in a concentration range of 20 - 40 mug/mL.However, 3-O-(2', 2'-dimethylsuccinyl)-betulinic acid (2) was also found to be an inhibitor of HIV-induced membrane fusion with IC_<100> value of 20mutg/mL, though it displayed significantly lower anti-HIV activity than foregoing compounds with ED_<50> value of 2.7 muM and T.l. values of 5.9. This observation indicated that another mechanism(s) of action other than inhibition of syncytia formation could be involved in the anti-HIV activity shown by these compounds. Compound 3 was not active in the MAGI assay, suggesting that it inhibit HIV replication at a late stage of the viral life cycle, such as after viral protein synthesis.On the other hand, oleanolic acid was identified as an anti-HIV principle from several plants, including Rosa woodsii (leaves), Prosopis glandulosa (leaves and twigs), Phorodendron juniperinum (whole plant), Syzygium claviflorum (leaves), Hyptis capitata (whole plant), and Terniroemia gymnanthera (aerial part). It inhibited HIV- 1 replication in acutely infected H9 cells with an EC_<50> value of 3.7 muM, and inhibited H9 cell growth with an IC_<50> value of 47.8 muM [therapeutic index (T.I.) 12.8]. Pomolic acid, isolated from R.woodsii and H.capitata, was also identified as an anti-HIV agent (EC_<50> 3.0 muM, T.I.16.6). Although ursolic acid did show anti-HIV activity (EC_<50> 4.4 muM), it was slightly toxic (IC_<50> 14.3 muM, T.I.3.3). Based on these results, we examined anti-HLV the activity of oleanolic acid- or pomolic acid-related triterpenes isolated from several plants. In addition, we previously demonstrated that derivatives of betulinic acid, isolated from the leaves of S.claviflum as an anti-HIV principle, exhibited extreme potent anti-HIV activity. Accordingly, we prepared derivatives of oleanolic acid, and evaluated their anti-HIV activity. Among the oleanolic acid derivatives, 3-O-(3', 3'-dimethylsuccinyl)-oleanolic acid demonstrated most potent anti-HIV activity, with an EC_<50> value of 0.00086 muM and a T.I.value of 22,400.Though ursolic acid is slightly toxic against H9 cells, it shows similar level of anti-HIV acitivity to that of oleanolic acid. In addition, the structure of ursolic acid is closely correlated with oleanolic acid and betulinic acid, hence it is regarded as a potential lead for anti-HIV agent. Therefore, similar modification of ursolic acid, and evaluated their anti-HIV activities. 3-O-diglycoryl-uroslic acid demonstarated relatively potent anti-HIV activity with an EC_<50> value of 0.31 muM and a T.I.value of 155.5. Hoever, anti-HJV activities for other derivatives, including 3,3-dimethysuccinyl derivative, were not potent as corresponding betulinic acid and oleanolic acid derivatives. Less

基于Betulinic Acid（1）和二氢苯甲酸（9）被鉴定为抗Hiv-Hiv Agents，对一系列卢潘烷型三萜酸衍生物进行了合成并评估其对急性感染的H9细胞中HIV-1复制的抑制活性。在衍生物中，3-o-（3'，3'二甲基二甲基二甲基） - β-竞争酸（3）和3-o-（3'，3'，3'-二甲基 - 核酸） - 二羟基苯甲酸（11）都表现出具有极具潜在抑制活性，具有EC_ <0.000035 mum.thebore comportive ec_ <0.00035 mum bum theecried except in。 >> 20,000和> 14,000。 3-o-（3'，3'二甲基戊二烯酸和 - 二氢苯甲酸（12），3-O-二甲基丙基 - 不能乙酸和-Dihydrobetulinic酸和-Dihyrobetulinic酸，以及3-o-甲状腺素酸也可能抑制0.0.0.0.0.0. 000均可抑制0. 50. 50. 50. 50. 50. 50. 50. 50. 502妈妈和T.I.Values从1,017到2,344。此外，化合物1 1和12在单核中也对HLV复制也有效……更多的TE细胞系和N外周血单核细胞（PBMC）。我们的体外测定法表明，这些化合物不是HIV -1逆转录酶的抑制剂，而它们完全抑制合成症的浓度范围为20-40 mug/ml。然而，还发现3-O-（2'，2'，2'二甲基二甲基） - 葡萄酸（2）是HIV诱导的膜融合的抑制剂，IC_ <100>值为20mmutg/ml，尽管它显示出比ED_ <50> <50> <50> 7 mum和T.7 mum和T.7 mum and t.7 mumm and and t.7 mum and t.7 mum and t.7 mum and and and t.7 mumm and t.7 mumm and t.7 mumm and t.7 mumm and t.7 amm and。值5.9。该观察结果表明，除了抑制合胞体形成以外的其他作用机制可能参与了这些化合物所示的抗HIV活性。 Compound 3 was not active in the MAGI assay, suggesting that it inhibit HIV replication at a late stage of the viral life cycle, such as after viral protein synthesis.On the other hand, oleanolic acid was identified as an anti-HIV principle from several plants, including Rosa woodsii (leaves), Prosopis glandulosa (leaves and twigs), Phorodendron juniperinum (whole plant), Syzygium claviflorum（叶子），Hyptis Capitata（整个植物）和Ternioemia Gymnanthera（空中零件）。它抑制了EC_ <50>值为3.7 MUM的急性感染的H9细胞中的HIV-1复制，并以IC_ <50>值为47.8 MUM [治疗指数（T.I.）12.8]抑制H9细胞生长。从r.woodii和H. capitata中分离出的荷酸也被确定为抗HIV剂（EC_ <50> 3.0 Mum，T.I.16.6）。尽管ursolic酸确实表现出抗HIV活性（EC_ <50> 4.4 MUM），但它有点毒（IC_ <50> 14.3 Mum，T.I.3.3）。基于这些结果，我们检查了抗HLV，从几个植物中分离出的灰甲醇酸或氯酸相关的三萜的活性。此外，我们先前证明了从链球菌的叶片中分离出的β叶酸的衍生物作为抗HIV原理，暴露了极端势抗HIV活性。根据链球菌作为抗HIV原理的叶子，暴露了极端的潜在抗HIV活性。在丁香酸衍生物中，3-O-（3'，3'-二甲基酸氨基甲醇） - 稀醇表现出最有效的抗HIV活性，EC_ <50>值为0.00086 MUM，T.I.VALUE和22,400的VALUE，URSOCIC酸略有毒性。另外，尿酸的结构与丁香酸和β酸性密切相关，因此被认为是抗HIV剂的潜在铅。因此，类似的ursolic酸修饰，并评估了它们的抗HIV活性。 3-O-diglycoryl-脉络酸分别表现出潜在的抗HIV活性，其EC_ <50>值为0.31 MUM和155.5的T.I. Value。 hoever，其他衍生物的抗HJV活性，包括3,3-二甲基二甲基衍生物，无潜力作为相应的β酸和稀醇衍生物。较少的