治疗用A型肉毒结晶毒素的研制及实验动物模型的建立

A型肉毒梭菌结晶毒素是以酸等电点沉淀,磷酸盐提取,核糖核酸酶处理,DEAE-A_(50)离子交换层析,硫酸铵外液透析制备的。毒力强,性能稳定,冻干制品宜于长期保存。经生化学、免疫学试验表明,毒素系神经毒素和血凝素的复合体,纯度高达2.5～2.6×10~7LD_(50)(小白鼠)/mgpr。给实验动物—恒河猴眼外肌注射能致人工斜视再给猴同眼的拮抗肌注射毒素,斜视得以矫正。可望成为一种治疗眼、面等神经、肌肉痉挛性疾病的安全,有效制剂。

Preparation of Type A Botulinum Crystalline Toxin for Therapeutics and Establishment of an Experimental Animal Model

A protocol for preparing type A botulinum crystalline toxin was described. The procedures include; acid precipitation, extraction with phosphate buffer, RNase treatment, DEAE-A50 ionexchange chromatography and dialysis against concentrated solution of Amso 4. These toxic crystals, which retained hemagglutinin, not only had high purity (2.5~2.6×10~7 LD50/mg. protein, high molecular weight substance account for 98% of total protein), but also showed good stability. After lyophilization, they could be preserved for a long term.Animal model (monkey)experiments indicated that the LD50 of monkey was close to 50 LD50 (mouse)/Kg. However, 0.2 LD50 (mouse)/Kg was effective for making an artificial strabismus and correcting them to normal as well, when it had been exactly injected into antagonistic extraocular muscle in proper order and time. Though an amount of toxin, 10 LD 50(mouse)/Kg, was given to monkeys, they were still healthy and did not occur any systemic symptom of botulism. So it could be concluded that the products were good in quality, if a suitable amount were applied under control, they should be very effective preparation for treating some diseases of muscular spasm in human.