Catalysis

Hydroxylation

Ecology

Biology

Steroid

Monooxygenase

Telecommunications

Biochemistry

Core (optical fiber)

Computer science

Cytochrome P450

Substrate (aquarium)

Stereochemistry

Combinatorial chemistry

Organic chemistry

Chemical synthesis

In vitro

Enzyme

Pharmacology

Hormone

Medicine

Stereoselectivity

Chemistry

ACS Catalysis

The synthesis of steroid drugs by multistage modifications of the steroidal core is challenging since site-specific and selective modification is essentially required, which is often difficult or complicated for chemocatalysis. For example, the synthesis of Trenbolone (3), a versatile anabolic–androgenic steroid, relies on a four-step chemical procedure on its core modifications of estra-4,9-diene-3,17-dione (1). Here, we have designed a two-step chemoenzymatic strategy that includes a biocatalytic one-pot C11-hydroxylation/17β-ketoreduction of 1 with a computationally designed P450 monooxygenase and an appropriate 17-ketosteroid reductase to generate 11α-OH-9(10)-dehydronandrolone (2a) as an intermediate followed by chemical dehydration to introduce the double bond at carbons 11 and 12 with the formation of Trenbolone (3). To obtain a highly active and C11-selective enzyme, molecular dynamics simulations were performed, uncovering a crucial role of water molecules for substrate recognition and targeted hydroxylation of steroids. Moreover, Trenbolone is further subjected to esterification to produce Trenbolone acetate (9) that has been widely used in veterinary medicine. Finally, our approach enables the regio- and stereoselective synthesis of both steroid drugs 3 and 9 on a (nearly) gram scale with 83–91% isolated yields, showing great potential for industrial applications.

A Chemoenzymatic Strategy for the Synthesis of Steroid Drugs Enabled by P450 Monooxygenase-Mediated Steroidal Core Modification