The selective functionalization of aliphatic C-H bonds in complex organic molecules remains a most challenging problem in organic chemistry. Development of efficient methods to afford this transformation, in particular in the context of biologically active natural products, is bound to disclose unprecedented opportunities in the transformation of these molecules, thereby accelerating drug discovery efforts. In this application, we propose research aimed at developing powerful new methodologies to exploit P450 C-H oxidation catalysis and P450-mediated synthesis for selective transformation of unreactive aliphatic C-H bonds in complex natural products. To this end, efficient and time-effective strategies will be implemented to enable tailoring of the substrate reactivity, regio- an stereoselectivity of P450 monooxygenases and rapid generation of selective P450 catalysts for mediating functionalization of distinct unactivated aliphatic sites within a complex organic scaffold. At the basis of the proposed strategies there is a new enabling methodology to map the active site configuration in P450 monooxygenases and rapidly acquire information-rich functional profiles of these enzymes ('P450 fingerprints'). Systematic methods will be implemented to predict the reactivity of P450 variants via analysis of their fingerprints and to identify optimal mutagenesis schemes for altering the active site configuration in P450 monooxygenases without disrupting catalytic function. These new tools will be applied to obtain P450 catalysts with tailor-made regio- and stereoselectivity for hydroxylation of multiple aliphati positions in terpenes with clinically relevant anticancer and antimalarial activity. These efforts will unlock new avenues for late-stage transformation of these molecules in order to improve their pharmacological properties. By coupling selective P450-catalyzed aliphatic hydroxylations to chemical synthesis, novel and currently inaccessible derivatives of these natural products will be made available for activity evaluation. Through these studies, comprehensive structure-activity insights will be gleaned on these natural products, providing a basis for development of more potent antileukemic and antimalarial agents. Completion of the primary objectives of this application will result in a set of powerful and general strategies for P450 C-H oxidation catalyst development which will be readily applicable to a variety of other natural products and high- value compounds.

Public Health Relevance

This research will develop new methodologies to obtain selective P450 oxidation catalysts for functionalization of unreactive aliphatic C-H bonds in complex molecules. These methods will be directly relevant toward the transformation and diversification of therapeutically relevant natural products, which represents a challenging problem in medicinal chemistry.

National Institute of Health (NIH)
National Institute of General Medical Sciences (NIGMS)
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Synthetic and Biological Chemistry B Study Section (SBCB)
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Fabian, Miles
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University of Rochester
Schools of Arts and Sciences
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Alwaseem, Hanan; Frisch, Benjamin J; Fasan, Rudi (2018) Anticancer activity profiling of parthenolide analogs generated via P450-mediated chemoenzymatic synthesis. Bioorg Med Chem 26:1365-1373
Saab-Rincón, Gloria; Alwaseem, Hanan; Guzmán-Luna, Valeria et al. (2018) Stabilization of the Reductase Domain in the Catalytically Self-Sufficient Cytochrome P450BM3 by Consensus-Guided Mutagenesis. Chembiochem 19:622-632
Moore, Eric J; Steck, Viktoria; Bajaj, Priyanka et al. (2018) Chemoselective Cyclopropanation over Carbene Y-H Insertion Catalyzed by an Engineered Carbene Transferase. J Org Chem 83:7480-7490
Wei, Yang; Tinoco, Antonio; Steck, Viktoria et al. (2018) Cyclopropanations via Heme Carbenes: Basic Mechanism and Effects of Carbene Substituent, Protein Axial Ligand, and Porphyrin Substitution. J Am Chem Soc 140:1649-1662
Brandenberg, Oliver F; Fasan, Rudi; Arnold, Frances H (2017) Exploiting and engineering hemoproteins for abiological carbene and nitrene transfer reactions. Curr Opin Biotechnol 47:102-111
Sreenilayam, Gopeekrishnan; Moore, Eric J; Steck, Viktoria et al. (2017) Metal Substitution Modulates the Reactivity and Extends the Reaction Scope of Myoglobin Carbene Transfer Catalysts. Adv Synth Catal 359:2076-2089
Sreenilayam, Gopeekrishnan; Moore, Eric J; Steck, Viktoria et al. (2017) Stereoselective olefin cyclopropanation under aerobic conditions with an artificial enzyme incorporating an iron-chlorin e6 cofactor. ACS Catal 7:7629-7633
Moore, Eric J; Zorine, Dmitri; Hansen, William A et al. (2017) Enzyme stabilization via computationally guided protein stapling. Proc Natl Acad Sci U S A 114:12472-12477
Tinoco, Antonio; Steck, Viktoria; Tyagi, Vikas et al. (2017) Highly Diastereo- and Enantioselective Synthesis of Trifluoromethyl-Substituted Cyclopropanes via Myoglobin-Catalyzed Transfer of Trifluoromethylcarbene. J Am Chem Soc 139:5293-5296
Tyagi, Vikas; Alwaseem, Hanan; O'Dwyer, Kristen M et al. (2016) Chemoenzymatic synthesis and antileukemic activity of novel C9- and C14-functionalized parthenolide analogs. Bioorg Med Chem 24:3876-3886

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