Moeremans M, De Raeymaeker M, Daneels G, De Brabander M, Aerts F, Janssen C, Andries K
Janssen Research Foundation, Beerse, Belgium.
Antimicrob Agents Chemother. 1992 Feb;36(2):417-24. doi: 10.1128/AAC.36.2.417.
The binding of the antiviral compound R 61837 to human rhinovirus 9 (HRV 9) was studied quantitatively and compared with binding of R 61837 to HRV 9H, a semiresistant variant. For both strains, radiolabelled R 61387 bound to native particles only. The Kd values obtained by Scatchard analysis of saturation binding data were 37 nM for HRV 9 and 172 nM for HRV 9H, whereas the concentrations resulting in a 50% reduction of cytopathic effect were 42 nM and 840 nM, respectively. Reversibility experiments showed that 65% of the compound could be extracted with chloroform from HRV 9H but less than 5% could be extracted from HRV 9. Dissociation studies demonstrated that in the presence of excess unlabelled compound, the half-lives of the virus compound complex HRV 9 and HRV 9H were 385 and 15 min, respectively. The effect of this antirhinoviral compound on the formation of subviral particles induced by low pH or heat was also investigated. Rate zonal centrifugation experiments using [35S]methionine-labelled HRV 9 showed that binding of R 61837 protected the virus against heat (56 degrees C) and acid (pH 5.0) and that at the same concentration of R 61837 the semiresistant strain was stabilized to a lesser extent. This observation was confirmed immunochemically with nonneutralizing and neutralizing monoclonal antibodies. Both 80S and 130S subviral particles have C antigenic determinants, whereas native particles (150S) have been designated D. R 61837 prevented the switch from D to C antigenicity which can be induced by exposure of rhinoviruses to mild denaturing conditions. These findings indicate that the compound is able to prevent a conformational change of the capsid which may be a prerequisite for infection.
对抗病毒化合物R 61837与人类鼻病毒9型（HRV 9）的结合进行了定量研究，并与R 61837与半抗性变体HRV 9H的结合进行了比较。对于这两种毒株，放射性标记的R 61387仅与天然颗粒结合。通过对饱和结合数据进行Scatchard分析得到的Kd值，HRV 9为37 nM，HRV 9H为172 nM，而导致细胞病变效应降低50%的浓度分别为42 nM和840 nM。可逆性实验表明，65%的化合物可从HRV 9H中用氯仿提取，但从HRV 9中提取的不到5%。解离研究表明，在存在过量未标记化合物的情况下，病毒-化合物复合物HRV 9和HRV 9H的半衰期分别为385分钟和15分钟。还研究了这种抗鼻病毒化合物对低pH或热诱导的亚病毒颗粒形成的影响。使用[35S]甲硫氨酸标记的HRV 9进行的速率区带离心实验表明，R 61837的结合可保护病毒免受热（56℃）和酸（pH 5.0）的影响，并且在相同浓度的R 61837下，半抗性毒株的稳定性较低。用非中和和中和单克隆抗体进行的免疫化学实验证实了这一观察结果。80S和130S亚病毒颗粒都有C抗原决定簇，而天然颗粒（150S）被指定为D。R 61837可防止鼻病毒暴露于温和变性条件下诱导的从D抗原性向C抗原性的转变。这些发现表明该化合物能够防止衣壳的构象变化，而这可能是感染的先决条件。
