PolyCYPs+ kit instructions
PolyCYPs+ screening kit
Kit contains 18 PolyCYPs, AOX and FMO3
PolyCYPs+ Diversification kit
Kit contains 8 PolyCYPs isoforms
PolyCYPs+ Scale-up kit
Resupply of hit isoforms
PolyCYPs+ incubation conditions
Incubation conditions are important when conducting PolyCYPs reactions to ensure the best % conversions. Examples of suitable incubation conditions are given on page 3 of the instructions. We use a standard 150rpm on a shaker with a 5cm orbit for our PolyCYPs incubations. Other shaker types may have a different orbit which will impact oxygen transfer. Use this handy calculator for determining the best rpm to use for your shaker. You will need to know the orbit of your shaker in cm in order to use the calculator. The calculator works for most types of shakers but please contact us for more advice if your shaker has a very low orbit.
Recommendations and tips
Introducing oxygen into a drug candidate late in the optimisation process has several applications including exploration of SAR (structure-activity relationships) and the ability to access derivatives that may possess superior properties such as improved metabolic stability and LLE (ligand-lipophilicity efficiency). Biocatalysis can provide access to chemical space in a complementary manner to chemical synthesis and provide a “one-experiment” solution to accessing multiple derivatives in parallel. This poster illustrates the application of a new biocatalysis kit, PolyCYPs®, to enable parallel synthesis of hydroxylated derivatives of drugs.
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.
Hypha’s PolyCYPs kits are in routine use by pharma and agchem companies for producing human and other mammalian metabolites. One application involves use of PolyCYPs for creating radiolabelled metabolites for direct comparison with the radio profiles from mass balance and distribution study samples, necessary for regulatory filing. PolyCYPs provides a clean route for scalable access to more of the CYP-derived metabolites observed in these matrices, for definitive MetID and any tox studies deemed necessary. This is especially useful where low concentrations or unstable metabolites in the mass balance sample make structural identification difficult.
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Hypha Discovery is a UK-based CRO supporting pharmaceutical and agrochemical companies worldwide through the production of metabolites and new derivatives of drugs and agrochemicals in discovery and development.