Production of Human and Novel Non-mammalian Metabolites of Cyclosporin A
It is well known that metabolites of drug compounds may have different efficacy and properties to that of the parent compound. Investigation and production of these metabolites is critical for exploration and understanding of SAR, and to ensure thorough patent coverage. Metabolites can also be sourced to create antibodies used in development of assays for therapeutic drug monitoring.
Hypha has the ability to create both mammalian and microbial metabolites of drug candidates at scale, in order to permit characterization and assessment towards full SAR and patent coverage. All of the main human metabolites of Cyclosporin A were observed through biocatalysis using Hypha’s microbial panel, which resulted in selective hydroxylation as illustrated. Additionally, novel microbial-derived metabolites were observed and isolated. Selected PolyCYPs enzymes are also capable of generating metabolites of cyclosporin.
Hypha’s microbial biocatalysis process is effective at generating metabolites at up to gram scale. Through Hypha and Selcia’s partnership, [13C], [14C], [2H], [3H] and [15N]-labelled metabolites can be accessed to support regulatory, development or research projects in the pharma and crop protection industries. Hypha establishes optimized processes using unlabelled or stable labelled parent substrates, which can then be transferred to Selcia’s state-of-the-art radiochemistry labs for the production of radiolabelled metabolites.
In this case study at least 2 mg of a monohydroxylated metabolite (M4), originally observed in rat liver microsomes, was required by a US pharma company. Screening of the parent compound against 22 PolyCYPs enzymes and 23 microbes revealed the production of two main monohydroxylated products, one of which was M4. M4 was best produced by PolyCYPs 152 and 359, as well as by bacterial species 45. The other monohydroxylated metabolite was produced by a different PolyCYPs isoform, (PolyCYP 350) and bacterial species 1.
Metabolite identification is an integral part of both preclinical and clinical drug discovery and development. Synthesis of drug metabolites is often required to support definitive identification, preclinical safety studies and clinical trials.