Study On The Effect Of Target Tissue Interstitial Fluid Turnover Rate In The Difference Of Efficacy Of Three Anti-TNF-α Biologics Based On MPBPK Model

Objective: On the basis of the second-generation mPBPK model,the study aims to explore the effect of the interstitial fluid turnover rate of the target tissue and the inhibitory effect of the drug on the pathogenic target tumor necrosis factor-α(TNF-α),combined with in vivo optical imaging technology of mice,the elimination of TNF-α at target tissues on the pathological model animals(acute paw edema inflammation model)in vivo to study.Furthermore,we systematically evaluate the role of interstitial fluid turnover rate in the difference of efficacy of three anti-TNF-α biological agents Adalimumab(Ada),Infliximab(Inf)and Etanertcept(Eta)in the treatment of autoimmune diseases.Methods: The foot swelling and inflammation model of C57BL/6 mice induced by carrageenan(CA)was established.The Alexa Fluor 680 labeled bovine serum albumin(BSA)combined with small animal real-time in vivo imaging technology was used to evaluate the difference in lymph flow velocity at the target organs of the model mice.Simultaneously,the elimination of TNF-α labeled by Dylight 680 in vivo was quantitatively analyzed by small animal real-time in vivo imager,and the effect of intercellular fluid renewal rate on the inhibition of three TNF-α antagonists(Ada,Inf and Eta)on TNF-α target in vivo was evaluated.Finally,the pharmacokinetic behavior of three kinds of TNF-α antagonists in mice with foot swelling and inflammation model was evaluated,and the optimized mPBPK model was used to fit the real in vivo data of mice.Results: C57BL/6 mice reached the peak of swelling 4 h after injection of 1% CA,and the inflammatory symptoms lasted until 7 h after modeling.After subcutaneous injection of 1μL Alexa Fluor 680 labeled BSA on the left hind paw of C57BL/6 mice in the model group and control group,the intercellular fluid turnover rate at the foot of the control group was0.32±0.02 and the negative control group was 0.31±0.02,which was not significantly different from the control group.The renewal rate of intercellular fluid in the model group was 0.21±0.02,which was significantly lower than that of the control group(P=0.0009).The results showed that CA modeling significantly reduced the renewal rate of intercellular fluid at the foot.The results of surface plasmonic resonance showed that Dy Light 680 did not affect the affinity of the three drugs to TNF-α.There was no significant difference in AUC between model group and control group after subcutaneous injection of 1 μL Dy Light680 labeled TNF-α in left hind paw paw of C57BL/6 mice and after subcutaneous injection of Ada and Inf.But after subcutaneous injection of Eta,there was a significant difference between the AUC of the model group and the control group(P=0.02),indicating that the reduction of intercellular fluid turnover rate in the model group had a significant effect on the elimination of TNF-α in the Eta administration group.In vivo pharmacokinetic curves of Ada,Inf and Eta were fitted with the redetermined binding kinetic parameters,and the fitting degree was good.Conclusion: Based on the second-generation mPBPK model,this study established a mice foot swelling inflammation model,using Alexa Fluor 680 labeled BSA combined with small animal real-time in vivo imaging technology to systematically evaluate the difference in intercellular fluid turnover rate at target organs of model mice.Through the in vivo elimination of TNF-α in model mice,it is found that when the intercellular fluid turnover rate at the target organ changes significantly,there is no significant difference between the efficacy of Ada and Inf,while the efficacy of Eta can be significant difference.The results of this study further confirmed the applicability of the second-generation mPBPK model in the pharmacodynamic evaluation of macromolecular antibody drugs.