Preformulation studies of thymopentin: analytical method development, physicochemical properties, kinetic degradation...

Thymopentin (TP5) is a synthetic pentapeptide with immunomodulatory properties. Given the previously described poor absorption of TP5, preformulation data is required to support effective formulation development. In this manuscript, an analytical method of TP5 was developed and validated to determine the aqueous solubility, stability, and Log P of TP5. Thermal properties were investigated, and chemical, physical and enzymatic degradation were evaluated. TP5 was informed to load in a microemulsion (ME) system according to the preformulation parameters and characterized for rheological behavior, droplet size, morphology and in vitro drug release. TP5 displayed high aqueous solubility (294.3 mg/mL), low Log P (-4.2) and 2% water content with a melting temperature of 193 °C. TP5 degraded rapidly in alkaline conditions, at elevated temperature, in oxidizing agents, and with UV exposure, however TP5 had a longer half-life in acidic conditions. The fastest enzymatic degradation was with Trypsin (half-life 6.3 h) compared with other digestive enzymes. The different degradation pathways followed first-order kinetics, and half-lives were obtained from the kinetic studies. The TP5 loaded ME exhibited a droplet size of 143 ± 35 nm with a Higuchi-model fitted sustained release profile for 24 h. These data justify and support the design of formulations to stabilize and enhance the absorption of TP5, with a ME formulation demonstrated.