Evaluation of Novel Phytobiologics as Potential Therapeutic Agents for Rheumatoid Arthritis

Abstract

Rheumatoid arthritis is an autoimmune, chronic inflammatory condition of the joints characterized by articular destruction and related extraarticular manifestations. Rheumatoid arthritis fibroblast like synoviocytes cells (RAFLS) mediate bone and cartilage destruction. Fibroblast like synoviocytes in RA exhibit a higher invasive, migrative and proliferative potential as compared to healthy fibroblasts as a consequence of metabolic reprogramming to withstand the augmented demands of these cells in terms of energy. Enrichment of glutaminolysis and glycolysis are two frequent modalities to the pathology RA. With no cure at the moment, for the autoimmune musculoskeletal disorder, several therapeutic strategies including steroidal, non-steroidal (NSAIDs), and disease modifying drug (DMARDs) classes are being used for disease management depending upon severity and stage of the disease. Thus, plant derived compounds have been explored as potential alternative therapeutic regimens. Plants provide an abundant and cheap source of anti-rheumatic, anti-inflammatory and analgesic compounds, all of which can be processed into a potent therapeutic for RA. Dracaena (Sansevieria) trifasciata is a common ethnomedicinal plant that has been linked with the treatment of numerous inflammatory diseases. However, its therapeutic effects on the underlying metabolic reprogramming in RA principally remains untapped. Hence, the current thesis dissertation was undertaken to explore if Dracaena (Sansevieria) trifasciata derived biologics may prove as a promising treatment option against RA. In the first phase of the study computer aided drug designing (CADD) was used to discover extremely efficient inhibitors against both HK2 and GLS (also known as GLS1) that obeyed the rules of drug likeness and pharmacokinetics. The identified inhibitors were xxvi a subset of the library of phytobiologics, that was obtained from the analysis conducted using GC-MS on ethanolic and methanolic extracts of Dracaena (Sansevieria) trifasciata. Docking and molecular dynamics simulations elucidated five compounds had the synergistic potential of regulating the pathologically escalated glycolysis and glutaminolysis. Out of the five potential leads, phytobiologic 28MS (c28MS) had a similar proclivity for targeting both HK2 and GLS, thus it was ensued for further testing to verify its therapeutic efficacy. In the second phase of our study, findings from RNA seq study delineated that fibroblast were a potential reliable target for the synergistic suppression of glutaminolysis and glycolysis. Henceforth, c28MS was subjected to in-vitro testing on RAFLS. The metabolic profile of RAFLS cells determined by 1H magnetic resonance spectroscopy under glycolytic and glutaminolysis suppressive conditions revealed that c28MS effectively decreased the elevated glucose/lactate and glutamine/glutamate ratio. Functional analysis of RAFLS unraveled that the c28MS lessened the aggressiveness of the RAFLS by checking their soaring invasion, migration, and proliferation capacity. In the third phase of the study, c28MS significantly reduced the intensity of the induced arthritis in the K/BxN model post-treatment. Undertaken study revealed that c28MS had the potential for successful glycolytic and glutaminolytic inhibition. The novel phytobiological inhibitor can successfully complement the existing treatments for RA for an improved management of the disorder. Additionally, all the identified inhibitors must undergo structural optimization for efficacy and enhanced selectivity before future clinical studies.