The Establishment Of Anaphylactoid Reaction Animal Models And Application In The Detection Of The Anaphylactoid Reaction Induced By Traditional Chinese Medicine Injection

Traditional Chinese Medicine Injection (TCMI) is a new and specific dosage form of China with high bioavailability and immediate effects. TCMI has been widely used in the therapy of critical emergency, infectious diseases, cardiovascular cancer and other diseases. With TCMI widely used in the clinic, the occurrence of adverse drug reactions increased gradually, especially the immediate hypersensitivity reaction. Immediate hypersensitivity reaction included anaphylaxis and anaphylactoid reaction, both of which present similar symptoms, diagnosis and treatment methods, so, anaphylactoid reaction is diagnosis of anaphylaxis in the clinical usually.At present, there isn't any model and guideline for TCMI is used in drug pre-clinical safety evaluation, the anaphylactoid reaction of TCMI can't be evaluated effectively. So, the establishment of sensitive and reliable anaphylactoid reaction models is very important for the survival and development of TCMI. This work was carried out with in the Major scientific and technological specialized project for Significant New Formulation of New Drugs.Part I. Establishment and application of anaphylactoid reaction models in ratAim:To establish a rat model for evaluation of TCMI anaphylactoid reaction and apply in the detection of anaphylactoid reaction induced by Qingkailing Injection (QKLI) and Gegensu Injection (GGSI). Methods:Detect the sensitivity and reliability of mean arterial blood pressure (MABP) as a indicator of rat model for anaphylactoid reaction by injecting histamine, Compound 48/80, Tween 80, Cremophor EL (CrEL), Radio Contrast Medium (RCM, Ultravist and Diatrizoate), Vancomycin, respectively, intravenous (i.v.); We also determined the feasibility of plasma histamine and Complement Terminatio Complex (SC5b-9) as indicators of anaphylactoid reaction Vancomycin by injecting Compound 48/80, Tween 80 and CrEL; Finally, we applied this rat model in the detection of anaphylactoid reaction induced by QKLI and GGSI. Results:In rat model, MABP decreased significantly after i.v. injection of histamine, Compound 48/80, Ultravist, Diatrizoate and Vancomycin, but not significantly after the injection of Tween 80, CrEL; Plasma histamine level, but not SC5b-9, was increased significantly after i.v. injection of Compound 48/80, Tween 80 and CrEL; MABP of rat model was decreased, with a increasing of plasma histamine level, as a dose and dose rate dependence way while i.v. injection of QKLI; A dose and dose rate dependence MABP decreased with a increasing of SC5b-9 could be caused by GGSI. Conclusion:Rat model for anaphylactoid reaction evaluation has good sensitivity to RCM, Vancomycin and anaphylactoid mediators of TCMI; QKLI and GGSI could cause a dose and dose rate dependence anaphylactoid reaction.Part II. Establishment and application of anaphylactoid reaction models in Beagle dogAim:To establish a Beagle dog model for evaluation of TCMI anaphylactoid reaction and apply in the detection of anaphylactoid reaction induced by QKLI and GGSI. Methods:Detect the sensitivity and reliability of plasma histamine, immunoglobulin E (IgE), SC5b-9, MABP, Respiratory Frequency (RF) and Heart rate (HR) as indicators of Beagle dog model for anaphylactoid reaction by injecting histamine, Compound 48/80, Tween 80, CrEL, RCM, Vancomycin, respectively, i.v.; Finally, we applied this Beagle dog model in the detection of anaphylactoid reaction induced by QKLI and GGSI. Results:In Beagle dog model, plasma histamine level, but not IgE, was increased significantly after i.v. injection of Compound 48/80, Tween 80, CrEL, RCM and Vancomycin; in addition, MABP decreased significantly with a increasing of SC5b-9 after i.v. injection of Compound 48/80, Tween 80, CrEL and Diatrizoate; A fast clinical infusion rate (300 ml/h) of QKLI could cause a MABP significant decrease and plasma histamine level increase; Normal (90 ml/h) and fast infusion rate of GGSI could cause a MABP significant decrease, and plasma histamine and SC5b-9 level increase both. Conclusion:Beagle model for anaphylactoid reaction evaluation has good sensitivity to Tween 80, CrEL, RCM and complement activators of TCMI; QKLI and GGSI could cause a dose rate dependence anaphylactoid reaction; Anaphylactoid reaction caused by GGSI may involve complement activation.Part III. Application of cat safety test models in detection of anaphylactoid reactionAim:To apply a standard safety test guideline of 2005 Pharmacopeia in detection of anaphylactoid mediators of QKLI and GGSI. Methods:In accordance with the standard Pharmacopoeia methods, to detect the anaphylactoid mediators of QKLI and GGSI. Results:QKLI, but not GGSI, got a negative result in anaphylactoid mediators'evaluation. Conclusion:There are some anaphylactoid mediators in GGSI, which could cause anaphylactoid reaction.Part IV. Development and application of cutaneous vascular permeability anaphylactoid reaction models in miceAim:To establish a mice model for evaluation of TCMI anaphylactoid reaction and apply in the detection of anaphylactoid reaction induced by QKLI and GGSI. Methods:To analysis the accuracy of cutaneous vascular permeability test, and establish the minimum error method; To analysis the reasons of the occurrence of false positive results of cutaneous vascular permeability, and exclude the impact of false positive results; Detect the sensitivity and reliability of cutaneous vascular permeability anaphylactoid reaction in mice by injecting Compound 48/80, Tween 80, CrEL, RCM, Vancomycin, respectively, intracutaneous (i.c.) and subcutaneous (s.c); Finally, we applied this Beagle dog model in the detection of anaphylactoid reaction induced by QKLI and GGSI. Results:Tissue Leachate had the maximum absorption and sensitivity at 610 nm; by 15 min after injection of Evans Blue, injected agents s.c. could prevent the occurrence of false positive results; QKLI and GGSI could cause increase of vascular permeability both. Conclusion:Mice cutaneous vascular permeability anaphylactoid reaction model has a good has good sensitivity and the least occurrence of false positive results by a method of 15 min after injection of Evans Blue, injected agents s.c; QKLI and GGSI could cause anaphylactoid reaction.