Studies On The Huachansu Parenteral Emulsion

Huachansu is prepared by the traditional Chinese drug—the desiccated skin of toad(Bufo bufo gargarizans Cantor or Bufo melanostictus Schneid).The pharmacological experiments have testified that it has many pharmacological actions,such as antitumor effects,analgesic effects, antivirus effects,et al.And it has been applied in clinic generally.The component of Huachansu is very complex,hydrosoluble and liposoluble constituents all have some pharmacologic actions. According to the Chinese traditional medicine recipe of Huachansu injection,Huachansu emulsion would be prepared by the optimization of Chinese crud drug extraction processes and screening of emulsion formula and preparation techniques.We expect that prepared Huachansu emulsion could elevate the stability of drug,degrade the adverse reaction,and enhance the therapeutic effect.TLC method was used to identify 5-HT,and the accurate,stable and convenient method of the content determination was built on the method by which the content of 5-HT in injection was determined(WS₃-B-3045-98).The best extraction techniques of toad skin were gained by orthogonal experiment,when contents of 5-HT were as detection index.The result of verification experiment indicated it was efficient and reproducible.The influences of fatty oil,emulsifier,viscosity-increasing agent on the emulsion were studied by single factor test,appearance of emulsion,appearance after centrifugation and Ke as index.Then based on these we found the best formula by orthogonal design when refining soybean oil,emulsifier and 1,2-propylene glycol were as consideration factors,Ke as investigation index.Effects of emulsification temperature and time,ultrasonic intensity and time, nanomizer working pressure and cycle times were also investigated by single factor test,based on which the optimized preparation procedure was obtained.The microphotograph showed that most of emulsion drops were globular in shape.The physicochemical properties of HCSPE such as particle size distribution,Zeta potential,pH value and viscosity is(77.0±9.9)nm,-23.7 mV,5.31,3.922×10^-3Pa·s respectively,and the osmotic pressure was slightly higher than the osmotic pressure of human plasma.There was no creaming after HCSPE was centrifugated at speed of 4000 r·min^-1for 15 min. The results of influencing factor test showed that HCSPE is stable at 40℃,but it should be tightly preserved from light.The results of accelerating and long-term tests indicated that the physical and chemical stability of HCSPE were fine and in compliance with the requirement of preparation stability.HCSPE combinated with physiologic saline were stable for 2 hours,and with 5%glucose were stable for 4 hours.The acute toxicity test demonstrated that LD₅₀of HCSPE was 209.2μg/kg,95%confidence interval was 206.7～211.8μg/kg.The security experiment indicated that blood vessel irritant reaction was not obvious,systemic anaphylactic and hemolysis reaction was negative.The HCSPE(20,40,80μg/kg)had obvious antitumor effects on H₂₂,and HCSPE(40,80μg/kg)had activities of immunoregulation in test of Clearance of charcoal particles.It was shown that the writhing response induced by acetic acid was remarkably inhibited by HCSPE (20,40,80μg/kg).The pain thresholds in the hot-plate test were increased by HCSPE(40μg/kg) in mice.Taking one with another,the HCSPE pharmacodynamic action of antitumor effects in vivo,immunoregulation activities and analgesic effects is better than Huachansu injection.