Study About The Effect Of FK506 And MMF Combined Self-assembled Immunosuppressant Cocktails In Acute Rejection Of Liver Transplantation

After decades of development,liver transplantation has been widely used in the treatment of various end-stage hepatic and biliary diseases.The widespread application of liver transplantation is not only due to the advancement of surgical techniques,but also the development of immunosuppressive agents which are routine drugs for prevention and treatment of postoperative acute rejection.At present,the main immunosuppressant for post liver transplantation are: calcineurin inhibitors,metabolic inhibitors,mammalian rapamycin target inhibitors,glucocorticoids and various monoclonal or polyclonal antibodies.Among these immunosuppressants,tacrolimus(FK506),a calcineurin inhibitor,is the cornerstone of immunosuppressive therapy after liver transplantation.In order to prolong the life of transplanted liver,FK506 must be taken orally for a long term after liver transplantation.However,long-term use of FK506 can also bring various adverse reactions.For the purpose of reducing its adverse reactions and playing a synergistic role in treatment,FK506 is often used in combination with the antimetabolite drug Mycophenolate Mofetil(MMF)in clinical practice.But,these two immunosuppressive agents are low water solubility,poor bioavailability and targeting.Because of the low water solubility,they are mainly taken orally,and it is difficult to prepare injectable drugs for routine use,which also limits their clinical application.To address these issues,we used the amphiphilic nano drug carrier based on the prodrug to load both drugs for the first time in this study.In addition,we established the Self-Assembled Immunosuppressant Cocktails(SAIC)based on the strategy of polyunsaturated fatty acid synthesis prodrugs in previous research.At the beginning of this study,we first selected MMF as a prodrug based on the structural feature of FK506 and MMF molecules.Moreover,linolenic acid(LA),docosahexaenoic acid(DHA)and heptanoic acid(Hep)were selected to conjugate with MMF,according to our previous investigation.All of these materials can be used to synthesize prodrugs with MMF,and the three prodrug MMF can be self-assembled into immunosuppressant cocktails with FK506 in water.Next,we compared the distribution of the three SAIC in vivo.The comparison results showed that LA-SAIC were most distributed in liver,spleen,and lymph nodes,which are correlated to acute rejection after liver transplantation.Consequently,we chose LA-MMF and FK506 to self-assemble into a immunosuppressant cocktails using on model of acute rejection after liver transplantation in rats in the following study.In the comparison of LA-SAIC with conventional FK506 combined with MMF orally,the LA-SAIC can significantly prolong the survival time of rats after allograft liver transplantation.It was also found that the LA-SAIC significantly inhibited the exudation of lymphocytes and the proliferation of exudative cells around the vessels in transplanted livers.Principal component analysis and clustering analysis suggested that LA-SAIC system group profiles were homogeneous and stable.Moreover,comparison for every chemokines and cytokines revealed that LA-SAIC can significantly inhibit multiple chemokines and promoting T cell proliferation cytokines,and notablely increase cytokines derived from Th2 cells in transplanted liver.Principal component analysis for liver function profiles of each group demonstrated that the profiles of the LA-SAIC did not change significantly,and clustering analysis point out that liver synthesis function was more homogeneous among three group liver function profiles.After comparing all liver function individually,we found that liver synthesis function of the LA-SAIC group was evidently better than the other two groups.And the damage of hepatic and biliary duct in the LA-SAIC group were obviously less than the rejection group.To sum up,we used a fatty acid-modified prodrug strategy to construct a self-assembling FK506 and MMF combined SAIC in this study.And LA was selected as an ideal MMF prodrug synthesis material.After synthesis of LA-MMF,it was self-assembled with FK506 to compose a combined LA-SAIC.The drug-loading system can significantly prolong the survival time of allograft liver tranplantation rats,and inhibit the acute rejection in transplanted liver.These results show that the LA-SAIC has a good therapeutic effect and clinical practice prospects for acute rejection after liver transplantation.