Investigation of tetrazole derivatives synthesized by the new method activities on tyrosinase enzyme, and its use as an active ingredient in cream formulation

In this study, it was aimed to obtain a synthetic skin whitening and anti-blemish compound by a new synthesis method and to use it in cream formulation. The study was carried out in four stages. In the first stage, a molecular docking study was performed. Derivatives with potential to inhibit the tyrosinase enzyme were determined for the 5-substituted-1H-tetrazol derivatives planned to be synthesized. The second stage is the synthesis stage. Benzylidenemalononitrile (BMN) and 5-substituted-1H-tetrazol derivatives were synthesized by developing a new methodology under the influence of aluminum oxy-hydroxide-supported Pd nanoparticles (Pd/AlO(OH) NPs) using aryl aldehyde derivatives. 11 BMN derivatives were synthesized with quantitative yields in the range of 2-24 hours and 12 5-substituted-1H-tetrazol derivatives were synthesized with 60-95% yields in the range of 3-6 hours The structures of the compounds were characterised by spectroscopic methods such as 1H NMR, 13C NMR and FT-IR. In the third stage, the tyrosinase enzyme activities of 5-substituted-1H-tetrazol derivatives were examined in vitro. In silico tests (BOILED-Egg model and bioavailability radar) were performed for the molecule 3-(3,4-dichlorophenyl)-2-(1H-tetrazol-5-yl)acrylonitrile (35), which has the highest enzyme inhibition activity (IC50: 45,00±3,28 µM). In the fourth stage, the cream formulation was created with 3-(3,4-dichlorophenyl)-2-(1H-tetrazol-5-yl)acrylonitrile (35). Cosmetic product tests (microbiological, antimicrobial protective activity, physicochemical tests and antipigmentation in vitro product tests) were performed on the obtained cream formulation. According to the microbiological analysis results, the formulation was determined to comply with the provisions of the "Guide on Microbiological Control of Cosmetic Products". S. aureus, P. aeruginosa, E. coli and C. Albicans microorganisms were not detected in the cream formulation. It was determined that the total number of aerobic mesophilic microorganisms and total mold and yeast did not exceed 102 cfu/g. With the antimicrobial protective effectiveness test, it was determined that the cream formulation content complies with the ISO 11930 acceptance criteria (criterion A) against the growth of microorganisms (S. aureus, P. aeruginosa, E. coli, C. albicans ve A. brasiliesis). In the anti-pigmentation test, the relevant formulation was applied to the human epidermal melanocyte cell line at increasing concentrations (25, 50, 100 and 200 µg/mL). The cream formulation was found to inhibit melanin synthesis compared to the control group.