药物类过敏反应的临床前评价方法研究(Ⅱ)——大鼠皮肤类过敏试验

Methodology for preclinical assay of pseudoallergy of injectable drugs(Ⅱ)——rat model for assay of cutaneous pseudoallergy induced by injections

目的:建立适用于注射剂类过敏评价的动物模型。方法:大鼠静脉注射含0.6%伊文思蓝(EB)的生理盐水溶液,10 min后,背部皮内注射受试物50μL,20 min后,测量皮内蓝斑直径和伊文思蓝渗出量,以评价类过敏反应程度。结果:生理盐水注射液(NS)、5%葡萄糖注射液造成很小的皮内蓝斑。阳性对照物Compound 48/80可导致局部毛细血管通透性显著增高和水肿,从而引起较大的蓝斑和EB大量渗出。采用临床上可发生类过敏反应的多种中、西药注射剂验证了本模型和评价方法的可靠性和敏感性,检测结果与临床一致。结论:大鼠皮肤类过敏试验方法与临床一致性。该方法敏感、可靠,适宜于注射剂临床前类过敏反应评价、注射剂生产过程中的安全性检测、上市产品抽验、致敏原筛选、类过敏反应发生机制研究以及类过敏防治方法研究等。

Objective： To develop animal models and methodologies for assay of pseudoallergy induced by injectable drugs. Method： Rats cutaneous anaphylactoid reaction model was developed by intravenous injection of 0. 6% Evans blue（EB） followed by intracutaneous injection of test substance solutions 50 μL. Diameters of subcutaneous blue spots and EB exudation were assayed. Re- suit： Rat anaphylactoid reaction was characterized as vascular hyperpermeability which was measured by diameters of blue spots inside the skin and the EB exudation of the blue spots. Compound 48/80 caused severe bluing and EB exudation in the skin by inducing obvi- ous vascular hyperpcrmeability which indicated that it can induce rat skin pseudoallergy. Normal saline or 5% glucose injection showed no obvious reactions. The rat pseudoallergy model was validated by intracutaneous injections of western drug injections and Chinese medicine. Conclusion： Rats could be developed into skin pseudoallergy model for preclinical safety evaluation of injectable drugs. The pseudoallergy reaction in this model is of high clinic consistency, sensitivity, reproducibility, and maneuverability. The model is suit- able for the evaluation for pseudoallergy induced by injectable products prepared from Chinese materia medica. This model can also be used for safety assay and quality control in manufacturing process, spot checking of marketed products, screening of allergen as well as studying of pseudoallergy mechanism.