Preparation Of MTX-BBR Soluble Microneedles And Study On The Mechanism Of Action Of MTX-BBR Based On Network Pharmacology

ObjectiveRheumatoid arthritis(RA)is an autoimmune disease with complex pathogenesis and can not be completely cured at present.Methotrexate(MTX)can effectively inhibit the progression of RA.However,choosing oral methotrexate makes its bioavailability unstable and subcutaneous injection will cause pain and sharp fear.In order to make up for the shortcomings of traditional drug administration,there is an urgent need for an alternative drug administration.Transdermal administration is a suitable choice.Transdermal drug delivery system is a new way of drug delivery,such as microneedles(MNs)transdermal drug delivery,by which drugs are absorbed into human blood circulation through the skin and reach effective blood concentration to achieve disease treatment.MN is a patch containing micron needle arrays that can physically break the stratum corneum skin barrier.We can control the length of the tip and pierce the MN tips into the epidermis and dermis of the skin.The drug can stimulate the immune response through antigen-presenting cells such as epidermal Langerhans cells and dermal dendritic cells,and can be quickly absorbed into the blood through rich capillaries in the dermis.In this study,drug-loaded soluble MNs were designed and prepared,and RA was treated by percutaneous administration,thus avoiding the pain of subcutaneous injection in patients with oral tolerance.The adjuvant therapeutic effect of BBR on RA was studied by network pharmacology.Construct the link between BBR-related targets and RA-related genes,and provide biological activities and signal pathways related to RA that can be regulated by BBR.MethodKonjac glucomannan(KGM)drug-loaded microneedles,including MTX microneedles,MTX-berberine(BBR)compound microneedles and blank microneedles,were designed and prepared by micromolding method.The shape and mechanical properties of MNs were characterized,including microphotography,needle negative pressure test,in vivo skin dissolution test and tip drug content determination.Based on network pharmacology,the auxiliary role of BBR in the treatment of RA was studied,including the prediction of MTX and BBR target proteins by PharmMapper database,the construction of protein interaction network map and drug-gene-disease network map,and GO functional enrichment analysis and KEGG metabolic pathway enrichment analysis of key genes.ResultsKGM-soluble microneedles prepared by micromolding method:each microneedle has a flat backing and complete needle structure;each microneedle patch can load 0.4243mg ± 0.0553 mg MTX and 0.6056mg ± 0.1001 mg BBR;the microneedle has mechanical strength enough to penetrate the skin;when the microneedle is punctured,the rat shows good compliance and can dissolve more than 90%in 20 min.Based on the research method of network pharmacology,the protein interaction network map of MTX and BBR related targets includes 29 nodes and 220 connections.A total of 33 biological activities were screened by GO functional enrichment analysis,and 13 biological activities were identified by KEGG signal pathway enrichment analysis.ConclusionsThe results showed that the KGM-soluble microneedles prepared in this study had the ability of drug loading and transdermal delivery.Based on the study of network pharmacology,the first five targets with a large number of protein interaction networks related to MTX and BBR are Albumin,Heat Shock Protein 90 Alpha Family Class A Member 1,Estrogen receptor 1,Non-Receptor Tyrosine Kinase and Mitogen-Activated Protein Kinase 14,suggesting that these targets may be the main pharmacological targets of MTX and BBR in anti-rheumatoid arthritis.MTX and BBR may affect the progress of RA activity by jointly regulating steroid binding,nuclear receptor activity,ligandactivated transcription factor activity,nuclear steroid receptor activity,ATPase binding and other biological processes.MTX and BBR may affect the progression of RA activity by jointly regulating Estrogen signal pathway,VEGF signal pathway,Chemical carcinogenesis-receptor activation,prolactin signal pathway,fluid shear stress and atherosclerosis.In summary,this study basically confirmed the feasibility of soluble microneedle treatment of RA based on MTX-KGM,and provided the biological activity and signal pathway that can be regulated by BBR adjuvant therapy of RA.