Cushman M, Wang P L, Chang S H, Wild C, De Clercq E, Schols D, Goldman M E, Bowen J A
Department of Medicinal Chemistry and Pharmacognosy, School of Pharmacy and Pharmacal Sciences, Purdue University, West Lafayette, Indiana 47907.
J Med Chem. 1991 Jan;34(1):329-37. doi: 10.1021/jm00105a052.
Aurintricarboxylic acid (ATA) was fractionated by a combination of dialysis, ultrafiltration, and gel permeation chromatography. The number average and weight average molecular weights of the ATA fractions were determined by the universal calibration method. The sulfonic acid analogue of ATA was prepared and separated in high and low molecular weight fractions. The phosphonic acid analogue of ATA was also synthesized. All of the ATA fractions were tested for prevention of the cytopathic effect of HIV-1 and HIV-2 in MT-4 cell culture as well as against HIV-1 in CEM cell culture. The abilities of the fractions and analogues to inhibit syncytium formation between HIV-1- and HIV-2-infected HUT-78 cells and uninfected MOLT-4 cells were evaluated. In addition, the fractions and analogues were tested for cytotoxicity in mock-infected MT-4 cells, prevention of the binding of the OKT4A monoclonal antibody to the CD4 receptor, inhibition of the binding of anti-gp120 monoclonal antibody to gp120, inhibition of attachment of HIV-1 virions to MT-4 cells, and inhibition of HIV-1 reverse transcriptase. In all of these assays except cytotoxicity, there was a correlation of potency with molecular weight. The higher the molecular weight, the higher the activity. Several of the lower molecular weight fractions of ATA, which bound to gp120 but not to CD4, prevented HIV-1 and HIV-2 cytopathicity. A similar profile was observed for the phosphonic acid analogue of ATA and the lower molecular weight fraction of the sulfonic acid analogue. The results on the ATA fractions indicate that the binding of ATA to gp120 in the absence of CD4 binding is sufficient for anti-HIV activity. The active compounds bind more avidly to gp120 than to CD4. The anti-HIV activity of the ATA fractions is due to inhibition of virus binding due to an interference with the gp120-CD4 interaction.
通过透析、超滤和凝胶渗透色谱相结合的方法对金精三羧酸（ATA）进行分级分离。采用通用校准法测定ATA级分的数均分子量和重均分子量。制备了ATA的磺酸类似物，并将其分离为高分子量和低分子量级分。还合成了ATA的膦酸类似物。对所有ATA级分进行了测试，以检测其在MT-4细胞培养中预防HIV-1和HIV-2细胞病变效应的能力，以及在CEM细胞培养中对抗HIV-1的能力。评估了这些级分和类似物抑制HIV-1和HIV-2感染的HUT-78细胞与未感染的MOLT-4细胞之间合胞体形成的能力。此外，还测试了这些级分和类似物在模拟感染的MT-4细胞中的细胞毒性、预防OKT4A单克隆抗体与CD4受体结合的能力、抑制抗gp120单克隆抗体与gp120结合的能力、抑制HIV-1病毒粒子与MT-4细胞附着的能力以及抑制HIV-1逆转录酶的能力。在除细胞毒性外的所有这些试验中，活性与分子量之间存在相关性。分子量越高，活性越高。ATA的几个低分子量级分与gp120结合但不与CD4结合，可预防HIV-1和HIV-2的细胞病变。ATA的膦酸类似物和磺酸类似物的低分子量级分也观察到类似的情况。ATA级分的结果表明，在不结合CD4的情况下ATA与gp120的结合足以产生抗HIV活性。活性化合物与gp120的结合比与CD4的结合更紧密。ATA级分的抗HIV活性是由于干扰了gp120-CD4相互作用从而抑制了病毒结合。
