Novel variant for application as a prolonged release drug delivery system

A dissertation submitted to the Faculty of Health Sciences, University of the Witwatersrand, in fulfilment of the requirements for the degree of Master of Science in Medicine 2015The dissertation aims to discuss the disulphide and thiol chemistry for use in drug delivery. In particular it focuses on the use of the modified native ovalbumin polymer as a vehicle for the thiol containing captopril. The binding capabilities of thiols expand the area in which peptides and proteins can be used as potential therapeutic drug carriers. It is important that drug delivery systems enhance drug storage stability and in vivo particle stability while delivering the drug efficiently. As part of the developing novel drug delivery systems, thiol-based chemical reactions are distinctive role players in stabilizing disulphide bioconjugated nanostructures for use as efficient drug carrier vehicles in vivo. A review of the current approaches for designing, optimizing and functionalizing nanostructures and conjugates by thiol chemistry modifications was explored. Captopril (Cp) is an Angiotensin-Converting Enzyme (ACE) inhibitor, which acts as an anti-hypertensive, structurally contains a free reactive thiol that binds variably via the thiol/disulphide reaction. A single dose of captopril can regulate hypertension for up to eight hours and the duration of the antihypertensive action of a single dose of 35-75 mg would be taken at 8 hour intervals for 24 hours. Hence the necessities in developing a sustained controlled release ovalbumin carrier system to maintain relatively constant blood pressure levels for 24 hours. The research focused on the construction, characterization and optimization of the thiol conjugated complex for sustained oral drug delivery. The thiol/disulphide-functionalized captopril-ovalbumin conjugate complex was assessed in terms of the structural characteristics and the thiol-disulphide covalent substitution reaction. For analysis of the conjugation complex, the Fourier Transmission IR-spectroscopy (FTIR), H+ NMR and Differential Scanning Calorimetry (DSC) was performed and used to confirm conjugation. Preliminary studies focused on a comparative study of sodium alginate, polyvinyl alcohol and hydroxypropylmethylcellulose hydrogel formulations for the release testing and drug entrapment of the ovalbumin-captopril conjugate complex. Utilizing this data, a series of process variables were used to achieve an optimized formulation through a Box- Behnken statistical design. Furthermore the drug release profiles of the optimised formulation were then analyzed in vitro and in vivo. The captopril released from the formulation was high with a cumulative release of 82%. In vivo analysis was the final testing to verify the validity of the ovalbumin-captopril conjugate complex encapsulated in sodium alginate and utilized a pig model. Ultra Performance Liquid Chromatography (UPLC) blood analysis revealed increased blood levels of captopril (Cmax Cp=33.2ng/mL) in relation to conventional dosage forms validating prolonged (24 hour) site-specific release and increased bioavailability. In conclusion, our validated method was successfully applied to the pharmacokinetic studies of captopril in the blood plasma samples