Campbell K P, Sharp A, Strom M, Kahl S D
Proc Natl Acad Sci U S A. 1986 May;83(9):2792-6. doi: 10.1073/pnas.83.9.2792.
Antibodies with high affinity and specificity for the 1,4-dihydropyridine Ca2+-channel blockers have been produced in rabbits by immunization with dihydropyridine-protein conjugates. Anti-dihydropyridine antibodies were found to specifically bind [3H]nitrendipine, [3H]-nimodipine, [3H]nisoldipine, and [3H]PN 200-110 (all 1,4-dihydropyridine Ca2+-channel blockers) with high affinity, while [3H]verapamil, [3H]diltiazem, and [3H]trifluoperazine were not recognized. The average dissociation constant of the [3H]nitrendipine-antibody complex was 0.06 (+/- 0.02) X 10(-9) M for an antiserum studied in detail and ranged from 0.01 to 0.24 X 10(-9) M for all antisera. Inhibition of [3H]nitrendipine binding was specific for the 1,4-dihydropyridine Ca2+-channel modifiers and the concentrations required for half-maximal inhibition ranged between 0.25 and 0.90 nM. Structurally unrelated Ca2+-channel blockers, calmodulin antagonists, inactive metabolites of nitrendipine, and UV-inactivated nisoldipine did not modify [3H]nitrendipine binding to the anti-dihydropyridine antibodies. Dihydropyridines without a bulky substituent in the 4-position of the heterocycle were able to displace [3H]nitrendipine binding, but the concentrations required for half-maximal inhibition were greater than 800 nM. In summary, anti-dihydropyridine antibodies have been shown to have high affinity and specificity for the 1,4-dihydropyridine Ca2+-channel blockers and to exhibit dihydropyridine binding properties similar to the membrane receptor for the 1,4-dihydropyridine Ca2+-channel blockers.
通过用二氢吡啶 - 蛋白质偶联物免疫兔子，已产生了对1,4 - 二氢吡啶钙通道阻滞剂具有高亲和力和特异性的抗体。发现抗二氢吡啶抗体能以高亲和力特异性结合[3H]尼群地平、[3H]尼莫地平、[3H]尼索地平以及[3H]PN 200 - 110（均为1,4 - 二氢吡啶钙通道阻滞剂），而[3H]维拉帕米、[3H]地尔硫䓬和[3H]三氟拉嗪则未被识别。对于详细研究的一种抗血清，[3H]尼群地平 - 抗体复合物的平均解离常数为0.06（±0.02）×10⁻⁹ M，所有抗血清的解离常数范围为0.01至0.24×10⁻⁹ M。[3H]尼群地平结合的抑制作用对1,4 - 二氢吡啶钙通道调节剂具有特异性，半最大抑制所需浓度范围在0.25至0.90 nM之间。结构不相关的钙通道阻滞剂、钙调蛋白拮抗剂、尼群地平的无活性代谢物以及紫外线灭活的尼索地平均不改变[3H]尼群地平与抗二氢吡啶抗体的结合。杂环4位没有庞大取代基的二氢吡啶能够取代[3H]尼群地平的结合，但半最大抑制所需浓度大于800 nM。总之，已证明抗二氢吡啶抗体对1,4 - 二氢吡啶钙通道阻滞剂具有高亲和力和特异性，并表现出与1,4 - 二氢吡啶钙通道阻滞剂膜受体相似的二氢吡啶结合特性。
