Late Stage Functionalisation
Generation of multiple hydroxylated derivatives of Alkermes’ lead compounds using PolarExplorer
A set of 7 lead compounds were screened blind against 12 PolyCYPs isoforms and the reaction products analysed by LC-MS to identify multiple oxidised derivatives, with 3 to 7 +16 Da products generated per compound. Derivatives of compounds 1 and 5 were prioritised for scale-up.
For compound 1, four +16 Da derivatives were observed in the initial screen and PolyCYP isoforms 359 and 484 were selected as the best to scale-up to capture these derivatives. Following purification of the scaled-up reaction, 5 different products were separated at a 33% isolated yield, ranging in weight from 2.46 mg to 28.05 mg. Alkermes opted to perform structure elucidation themselves on the purified derivatives provided by Hypha, revealing two derivatives with a secondary aliphatic hydroxylation, one derivative with a tertiary aliphatic hydroxylation, one derivative with aromatic hydroxylation and one rearranged structure from a tertiary aliphatic hydroxylation.
The work done to create oxidised derivatives using PolarExplorer allowed Alkermes to enhance their late-stage functionalisation efforts on compounds containing a complex molecular core and to rapidly expand their SAR understanding within projects.
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.
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.
In this paper, authors from Hypha and Incyte Corporation discuss the impact and application of biotransformation of drugs by mammalian systems, microorganisms, and recombinant enzymes, covering active and reactive metabolites, the impact of the gut microbiome on metabolism, and how insights gained from biotransformation studies can influence drug design.
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The services conducted by Hypha allowed Alkermes to enhance our late stage functionalization efforts to rapidly expand our SAR understanding inside projects. In particular, the PolarExplorer work allowed facile generation of hydroxylated compounds possessing a complex molecular core. We look forward to engaging Hypha in the future because their team is professional, and their services complement our internal synthetic efforts for delivering new molecules.
Brian Aquila, Director of Medicinal Chemistry
Alkermes, MA, USA
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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.