Biotransformation of bromhexine by Cunninghamella elegans , C. echinulata and C. blakesleeana

Abstract

Fungi is a well-known model used to study drug metabolism and its production in in vitro condition. We aim to screen the most efficient strain of Cunninghamella sp. among C. elegans, C. echinulata and C. blakesleeana for bromhexine metabolites production. We characterized the metabolites produced using various analytical tools and compared them with mammalian metabolites in Rat liver microsomes (RLM). The metabolites were collected by two-stage fermentation of bromhexine with different strains of Cunninghamella sp. followed by extraction. Analysis was done by thin layer chromatography, high performance thin layer chromatography, Fourier transform infrared spectroscopy, high performance liquid chromatography and Liquid chromatography–mass spectrometry. The role of Cytochrome P3A4 (CYP3A4) enzymes in bromhexine metabolism was studied. Fungal incubates were spiked with reference standard – clarithromycin to confirm the role of CYP3A4 enzyme in bromhexine metabolism. Three metabolites appeared at 4.7, 5.5 and 6.4 min retention time in HPLC. Metabolites produced by C. elegans and RLM were concluded to be similar based on their retention time, peak area and peak response of 30.05%, 21.06%, 1.34%, and 47.66% of three metabolites and bromhexine in HPLC. The role of CYP3A4 enzyme in metabolism of bromhexine and the presence of these enzymes in Cunninghamella species was confirmed due to absence of peaks at 4.7, 5.4 and 6.7 min when RLM were incubated with a CYP3A4 enzyme inhibitor – clarithromycin.

Biotransformation of 4‐fluoro‐N‐(1‐{2‐[(propan‐2‐yl)phenoxy]ethyl}‐8‐azabicyclo[3.2.1]octan‐3‐yl)‐benzenesulfonamide, a novel potent 5‐HT7 receptor antagonist with antidepressant‐like and anxiolytic properties: In vitro and in silico approach

“…This is consistent with previous reports comparing xenobiotics biotransformation by Cunninghamella spp. and microsomes, which indicated that the generated metabolites are comparable . Moreover, our in silico data coincide with in vitro experiments, as three MetaSite metabolites of PZ‐1150 matched compounds identified in microsomal samples.…”

“…Additionally, various microbial systems employing bacteria, yeast, and fungi may constitute an alternative to the use of animal models in the study of NCEs metabolism . The zygomycete fungus Cunninghamella can perform both phase I and phase II biotransformation reactions and was shown to metabolize chemical compounds in a way similar to mammals …”

Investigating the ability of the microbial model Cunninghamella elegans for the metabolism of synthetic tryptamines

“…[2][3][4] Metabolism studies can involve different in vivo and in vitro methods. Synthetic tryptamines are sold as new psychoactive substances (NPS) because of their hallucinogenic effects.…”

“…It is capable of metabolizing phase I and phase II biotransformation reactions. 4,17,18 Using microbial models makes it possible to avoid animal and human studies in drug discovery and development processes. 15 It has the cytochrome P450 CYP509A1 enzyme 16 and can facilitate reactions catalyzed by the human CYP1A2, CYP2C9, CYP2C19, CYP2D6, and CYP3A4.…”

“…2 Research has shown that C. elegans can produce the mammalian metabolites of xenobiotic drugs such as meloxicam, 13 naproxen, 14 or amitriptyline. 4 Other advantages include low costs, easy handling, and scale-up capability. 4,17,18 Using microbial models makes it possible to avoid animal and human studies in drug discovery and development processes.…”