Colaborative Research on Anti-HIV Triterpenoids

基于白桦脂酸（1）和二氢白桦脂酸（9）被鉴定为抗HIV药物的事实，合成了一系列羽扇烷型三萜酸衍生物，并评价了它们在急性感染的H9细胞中对HIV-1复制的抑制活性。其中3-O-（3 '，3 '-二甲基琥珀酰基）-白桦脂酸（3）和3-O-（3 '，3 '-二甲基琥珀酰基）-二氢白桦脂酸（11）均表现出极强的抑制活性，其EC<50>值&lt; 0.00035 μ M，并显示出显著的体外治疗指数（TI）。值分别为&gt; 20，000和&gt; 14，000。3-O-（3 ′，3 ′-二甲基戊二酰基）-桦木酸和-二氢桦木酸（12）、3-O-二甘醇酰基-桦木酸和-二氢桦木酸和3-O-戊二酰基-桦木酸也是HIV复制的有效抑制剂，艾德_<50>值范围为0.01 - 0.0023 μ M，T.I.值范围为1，017 - 2，344。此外，化合物11和12也具有抗HLV复制的活性。 ...更多信息 te细胞系和外周血单核细胞（PBMC）。我们的体外试验表明，这些化合物不是HIV- 1逆转录酶的抑制剂，但它们在20 - 40 μ g/mL的浓度范围内完全抑制合胞体的形成。然而，3-O-（2 ′，2 ′-二甲基琥珀酰基）-桦木酸（2）也被发现是HIV-诱导的膜融合的抑制剂，IC_<100>值为20 μ g/mL，尽管它显示出比前述化合物明显更低的抗HIV活性，其艾德_<50>值为2.7 μ M，T.值为5.9。该观察结果表明，这些化合物显示的抗HIV活性可能涉及除抑制合胞体形成以外的另一种作用机制。化合物3在MAGI测定中没有活性，表明它在病毒生命周期的后期阶段抑制HIV复制，例如在病毒蛋白合成之后。另一方面，从几种植物中鉴定出油酸是抗HIV的主要成分，所述植物包括木蔷薇（Rosa woodsii）（叶）、腺牧豆（Prosopis glandulosa（叶和嫩枝）、Phorodendron juniperinum（整株）、Syzygium claviflorum（叶）、Hyptis capitata（整株）和Terniroemia gymnanthera（地上部分）。在急性感染的H9细胞中，它以3.7 μ M的EC_值抑制HIV- 1复制<50>，以47.8 μ M的IC_值抑制H9细胞生长<50>[治疗指数（T.I.）12.8]。从R.woodsii和H.capitata中分离的Pomolic acid也被鉴定为抗HIV剂（EC_<50>3.0 μ M，T.I.16.6）。虽然熊果酸确实显示出抗HIV活性（EC_<50>4.4 μ M），但它有轻微的毒性（IC_<50>14.3 μ M，T.I.3.3）。基于这些结果，我们研究了从几种植物中分离的与油酸或棕榈酸相关的三萜类化合物的抗HLV活性。此外，我们以前证明，白桦酸的衍生物，分离自S.claviflum的叶子作为抗HIV的原则，表现出极强的抗HIV活性。因此，我们制备了油酸衍生物，并评价了它们的抗HIV活性。其中3-O-（3 ′，3 ′-二甲基琥珀酰基）-齐墩果酸的抗HIV活性最强，EC值<50>为0.00086 μ M，T.I.值为22，400。熊果酸对H9细胞有一定的毒性，但其抗HIV活性与齐墩果酸相当。此外，熊果酸的结构与油酸和桦木酸密切相关，因此它被认为是抗HIV药物的潜在先导。因此，对熊果酸进行类似的修饰，并评价其抗HIV活性。3-O-二甘氨酰尿苷酸具有较强的抗HIV活性，EC<50>值为0.31 μ M，T.I.值为155.5。然而，其他衍生物，包括3，3-二甲基琥珀酰基衍生物的抗HJV活性不如相应的桦木酸和油酸衍生物。少

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