DEVELOPMENT OF DRUG BIOCONJUGATE USING LIGHT SUBUNIT MUSHROOM TYROSINASE (LSMT) FOR ORAL DRUG DELIVERY

• Main Author: Diana
• Format: Theses
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/44433
• Institution: Institut Teknologi Bandung

Peroral routes for systemic therapy provides ease and convenience of drug administration for patients. However, preparation for peroral therapy provides challenges in terms of manufacturing technology. To ensure the active compound reaches its receptor, the drug must be absorbed into the blood vessels. Drug absorbtion becomes constrained if the permeability is low leading to limited bioavailability. The use of lectin for glycotargeting is an approach to overcome problems in the delivery of compounds with low permeability. Interaction between lectin with several types of oligosaccharides present in cells on the surface of gastrointestinal wall could caused the lectin to be absorbed. Vast glycosylated areas within gastrointestinal tracts can be targeted for this purpose. LSMT (light chain subunit in the tetramer complex of tyrosinase enzyme Agaricus bisporus) has the ability to recognize specific group of sugar moieties, non-toxic, and nonimmunogenic. Formation of LSMT-drug bioconjugate was explored in this study to asses the ability of LSMT as a drug carrier using captopril as a drug model. The study begins with assessing solvent accessibility of cysteine residue (functional target candidate for bioconjugation) using ASAView and NetSurfP programs. In vitro accessibility of cystein was performed to determine free sulfhydryl using DTNB reagent. From the study, lysine is chosen as an active side of the reaction. Conjugate is formed with SMPT as a linker utilizing a reduced disulphide bond to release the bound drug. Optimum conditions currently found for conjugate formation is at 4 ° C for 24 hours for protein activation stage with SMPT and 48 hours for captopril binding stage with ratio of protein: SMPT = 1:10 and activated protein:captopril = 1:100. Conjugate substitution obtained under these conditions is between 1-2 mol of captopril per mole of LSMT. Conjugate formed was stable in simulated gastric and intestinal solutions. Furthermore, preliminary in vitro permeability study using Caco2 cells and ex vivo with non-everted gut sac method showed intact ability of LSMT to penetrate gastrointestinal wall.