Anti-allergic Mechanism And Transdermal Pharmacokinetics Of THCA354 Gel

Purposes:Allergic dermatosis has a high incidence in the world,and it is easy to relapse when it comes to allergens after cure.Although the drugs used clinically for the treatment of allergic skin diseases have certain curative effects,there are certain side effects in long-term use.(E)-phenoethyl 3-(3,5-two hydroxyl-4-isopropyl phenyl)acrylate(THCA354)is a new polyphenolic acrylic acid derivative synthesized by our research group,and is formulated as external gel by prescription research.In this study,we observed the effects of THCA354 gel on the hypersensitivity of type I and type IV allergic skin diseases,and explored the mechanism of THCA354 gels anti type IV hypersensitivity by using mouse type IV hypersensitivity reaction model.The skin pharmacokinetics of THCA354 gel was studied by microdialysis combined with HPLC-MS/MS,so as to lay the foundation for developing the preparation of allergic dermatosis.Methods:1.Guinea pigs' active skin allergy model,rat passive skin allergic reaction model and allergic contact dermatitis model of mice were established to investigate the antiallergic effect of THCA354 gel.2.RT-PCR,ELISA and flow cytometry were used to detect Th1,Th2,Th17 and regulatory T cell(Tregs)-associated cytokines and cell-level responses in type IV hypersensitivity mice to investigate the anti-allergic immune regulation mechanism of THCA354 gel.3.Establish a HPLC-MS/MS method for the determination of THCA354 concentration in miniature pig skin,and validate the method as a basis for THCA354 in microdialysis samples.4.Microdialysis samples were obtained by microdialysis technique,and the concentration of THCA354 in the skin samples was determined by HPLC-MS/MS,and the concentration-time curve was plotted to calculate the pharmacokinetic parameters.Result:1.THCA354 gel anti allergy research results showed that THCA354 gel can significantly inhibit type I and type IV hypersensitivity induced allergic dermatosis.2.The anti-allergic mechanism of THCA354 gel showed that THCA354 gel can reduce inflammatory infiltration and skin edema of ACD mouse ear tissue induced by DNFB compared with model group.Down-regulate the levels of Th1/Th17-related cytokines and nuclear transcription factors IFN-?,IL-17,IL-6,T-bet and ROR-?t,up-regulate the levels of Th2/Treg-related cytokines and nuclear transcription factors IL-4,Foxp3,IL-10,TGF-? and GATA-3,reduce the number of CD3+CD4+IFN-?+T cells and CD3+CD4+CD4+IL-17A+T cells and induce the production of CD4+CD25+Foxp3+T cells and CD3+CD4+IL-4+T cells.3.Establish and investigate HPLC-MS/MS analytical methods for determining THCA354 concentration.The results show that the method is linear,and the recovery,precision and stability are in line with the requirements of in vivo drug determination,and can be used for the determination of microdialysis samples.4.The percutaneous pharmacokinetic study of THCA354 gel showed that THCA354 was detected after 100~12h of transdermal administration,and the drug stayed in the dermis for a long time and could continue to exert therapeutic effects.Conclusion:THCA354 gel has significant anti allergic effects,and its anti allergic mechanism has found that THCA354 gel can effectively inhibit the inflammatory infiltration and the secretion and expression of various proinflammatory factors,and promote the production of anti inflammatory factors.It can play an immune therapeutic role by changing the immune balance of Th1 / Th2 / Th17 / Tregs.The pharmacokinetic study found that THCA354 gel was absorbed smoothly after transdermal administration.After about 480 min,the concentration of THCA354 in the skin of Bama miniature pigs reached a peak of 436.46±23.21 ng/mL.THCA354 was still detected in the skin after 7 h of drug removal,and stayed in the dermis for a long time to continue to exert therapeutic effects.