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PolyCYPs Kits for Late-Stage Oxidation

PolyCYPs Kits for Late-stage Oxidation
Easy-to-use enzyme system for the rapid oxidation of organic molecules to generate oxidised derivatives of drugs, agrochemicals and other small molecules

Kits containing PolyCYPs enzymes, effective for synthesising oxidised metabolites and  derivatives of drugs, are available for clients to use in their own labs. PolyCYPs enzymes are an effective tool for late-stage oxidation, providing an opportunity to screen for and scale-up series of multiple hydroxylated derivatives in parallel, including identification of active metabolites.

Benefits such as greater polarity, enhanced metabolic stability, improved PK properties, increased potency and greater selectivity are potentially achievable.

PolyCYPs enzymes are cloned from some of Hypha’s microbes talented at oxidising a wide variety of compounds. They are formatted into easy-to-use kits containing all reagents needed, including a 24-well reaction block and air permeable seal.

Screening Kits

There are two screening kit options – a 20 enzyme kit containing 18 PolyCYPs isoforms, aldehyde oxidase (AOX) and flavo-containing monooxygenase 3 (FMO3) and an 8-enzyme “Diversification kit” containing 8 promiscuous PolyCYPs enzymes.

Content of PolyCYPs oxidation screening kit

PolyCYPs screening kit

8 or 20 oxidising enzymes

Cofactor recycling system containing glucose-6-phosphate (G6P), nicotinamide adenine dinucleotide phosphate (NADP+), and glucose-6-phosphate-dehydrogenase (G6PDH)

Formulation reagent to aid the solubilisation of test compounds of poor aqueous solubility

Extra enzyme (PolyCYP 6) for use with bosentan control reaction

Positive control substrate (bosentan)

All reagents are provided as lyophilised powders in sealed vials together with a 24-well reaction plate and air-permeable seal.

Everything is included in the kit that you will need to perform the reaction in your lab, just add your compounds and water!

PolyCYPs scale-up

PolyCYP enzyme reactions are fully scalable from mg to gram quantities through a number of routes, either by resupply of lyophilised enzymes for mg scale production in house, or larger scale production at gram scale at Hypha, with optional purification and structure elucidation.

Multiple 10ml scale-up vials, suitable for converting 1mg of parent compound per vial, can be ordered off-the-shelf for any isoform for in-house scale-up in a kit format. Co-factors and formulant are also included, along with a 24-well block suitable for performing 4 x 2.5 ml reactions. Other incubation formats can also be used, as recommended in the kit instructions.

For larger quantities of any PolyCYPs products, we offer reactions undertaken at Hypha using cell paste from the recombinant E.coli or streptomycete clones containing the enzyme of interest.

Hypha can also undertake screening of drug compounds using PolyCYPs enzymes on behalf of a client in a service called PolarExplorer. We offer a flexible end-to-end service including scale-up, purification and structure elucidation. Alternatively, scale-up reactions can be conducted at Hypha and products provided to the client for purification and/or structure elucidation.

Why use PolyCYPs oxidation kits?

Quick access to multiple oxidised derivatives in drug in parallel

Comes with all reagents needed, including a 24-well reaction block and air permeable seal

Readily scalable - use in-house or outsource to Hypha

Simultaneous access to CYP metabolites

Applicable to broad structural types and complex molecules, including natural products and synthetic compounds


Explore our library of resources comprising brochures, case studies, posters and publications about the work we do.

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.

This poster illustrates the application of a new biocatalysis kit, PolyCYPs®, to enable scalable synthesis of CYP-derived metabolites of drugs. The PolyCYPs platform is comprised of a set of recombinant cytochrome P450 enzymes and redox partners cloned from some of the talented actinomycetes in Hypha’s biotransformation panel. Enzymes in the kit catalyze the oxidation of a wide variety of substrate types to generate multiple mammalian and microbial-derived CYP metabolites.

A cell-free kit of cytochrome P450 enzymes and ferredoxin/ferredoxin reductase redox partners, termed PolyCYPs®, is being developed for generating scalable quantities of oxidised metabolites. P450 cytochromes in the kit have been derived from some of Hypha’s most talented biotransforming bacteria and are capable of generating human and other mammalian metabolites of drug compounds.

Test tubes with dripper
251A3171-DE01-48EB-AA27-BD82BC5AC10DCreated with sketchtool.

“We would like to mention that it has been a pleasure to work with Hypha in this project. We are really satisfied with how the collaboration has progressed, reaching the defined goals and timelines in a very efficient manner. Access of hydroxylated compounds through PolyCYPs has saved our company time and resources. We are sure that we will have more opportunities to work together in the future.”

Program Leader

European Pharma Company

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