Acetazolamide Inhibits the Level of Tyrosinase and Melanin: An Enzyme Kinetic, In Vitro, In Vivo, and In Silico Studies

Abstract

Melanin is the major factor that determines skin color and protects from ultraviolet radiation. In present study we evaluated the anti-melanogenesis effect of acetazolamide (ACZ) using four different approaches: enzyme kinetic, in vitro, in vivo and in silico. ACZ demonstrated significant inhibitory activity (IC 7.895 ± 0.24 μm) against tyrosinase as compared to the standard drug kojic acid (IC 16.84 ± 0.64 μm) and kinetic analyses showed that ACZ is a non-competitive inhibitor without cytotoxic effect. In in vitro experiments, A375 human melanoma cells were treated with 20 or 40 μm of ACZ with or without 50 μm of l-DOPA. Western blot results showed that ACZ significantly (P < 0.05) decreased the expression level of tyrosinase at 40 μm. Zebrafish embryos were treated with 10, 20 or 40 μm of ACZ and of positive control kojic acid. At 72 h of treatment with ACZ and kojic acid, ACZ significantly (P < 0.001) decreased the embryos pigmentation to 40.8% of untreated embryos at the dose of 40 μm of ACZ while kojic acid decreased only 25.0% of pigmentation at the same dose of kojic acid. In silico docking were performed against tyrosinase using PyRx tool. Docking studies suggested that His244, Asn260 and His85 are the major interacting residues in the binding site of the protein. In conclusion, our results suggest that ACZ is a good candidate for the inhibition of melanin and it could be used as a lead for developing the drugs for hyperpigmentary disorders and skin whitening.

Novel 1,3‐oxazine‐tetrazole hybrids as mushroom tyrosinase inhibitors and free radical scavengers: Synthesis, kinetic mechanism, and molecular docking studies

“…A series of experiments were performed to determine the inhibition kinetics of 3d by following the already reported method. [ ] The inhibitor concentrations for 3d were 0.00, 0.019, and 0.038 μM. Substrate L‐DOPA concentration was between 0.125 and 2 mM in all kinetic studies.…”

Exploration of Mechanistic Insights of Acemetacin in Melanogenesis Through Zebrafish Model, Enzyme Kinetics, Molecular Docking and Simulation Approaches

“…The potency of ACE against tyrosinase was investigated by exactly following our published method in various articles [19][20][21]. In the first step, 20 nM of phosphate buffer at pH 6.8 was prepared, and 140 µL of the solution was transferred in each well of the assay plate.…”

Triclosan induces ROS‑dependent cell death and autophagy in A375 melanoma cells

“…Melanoma only represents a small subset of these tumors, yet it is the most common skin tumor type, with increasing incidence and mortality rates worldwide ( 1 , 2 ). Currently, the primary treatments of malignant melanoma are surgical excision, immunotherapy, adjuvant chemotherapy, targeted therapy drugs ( 3 ) and radiotherapies ( 2 , 4 ). However, these therapeutic strategies do not facilitate the current clinical practice requirements due to the high metastatic potential and drug resistance ( 5 , 6 ).…”