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KEY POINTS * Serine hydrolases are one of the largest and most diverse classes of enzymes found in eukaryotes and prokaryotes, including ∼240 members in humans. * Several clinically approved

drugs target serine hydrolases. Prominent among these therapeutics are inhibitors of thrombin, acetylcholinesterase and dipeptidyl peptidase 4 that are used to treat clotting disorders,

Alzheimer's disease-associated dementia and diabetes, respectively. * Many serine hydrolases have recently emerged as enzymes with therapeutic potential and are the focus of intense

inhibitor discovery efforts. * Compounds that act through covalent mechanisms have proved to be especially effective at selectively inhibiting serine hydrolases. Here, we highlight the

mechanism-based electrophiles that have successfully formed the basis of selective, _in vivo_-active inhibitors (including several approved drugs) and also review promising new chemotypes

that have recently been discovered. * Activity-based protein profiling has facilitated the discovery of dysregulated serine hydrolases in disease and has enabled the rapid development of

selective inhibitors for the functional characterization of these enzymes. ABSTRACT Serine hydrolases perform crucial roles in many biological processes, and several of these enzymes are

targets of approved drugs for indications such as type 2 diabetes, Alzheimer's disease and infectious diseases. Despite this, most of the human serine hydrolases (of which there are

more than 200) remain poorly characterized with respect to their physiological substrates and functions, and the vast majority lack selective, _in vivo_-active inhibitors. Here, we review

the current state of pharmacology for mammalian serine hydrolases, including marketed drugs, compounds that are under clinical investigation and selective inhibitors emerging from academic

probe development efforts. We also highlight recent methodological advances that have accelerated the rate of inhibitor discovery and optimization for serine hydrolases, which we anticipate

will aid in their biological characterization and, in some cases, therapeutic validation. Access through your institution Buy or subscribe This is a preview of subscription content, access

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UNDERSTANDINGS AND IMPLICATIONS FOR DRUG DISCOVERY Article Open access 07 May 2025 STRUCTURAL INSIGHTS INTO THE INHIBITION OF GLYCINE REUPTAKE Article 03 March 2021 IN SILICO DESIGN OF NOVEL

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thank the Cravatt laboratory for helpful discussions. This work was supported by grants from the US National Institutes of Health (DA025285, GM090294, CA132630, DA017259, DA009789 and

CA087660), the National Science Foundation (predoctoral fellowship to D.A.B.), the California Breast Cancer Research Program (predoctoral fellowship to D.A.B.), and The Skaggs Institute for

Chemical Biology. AUTHOR INFORMATION AUTHORS AND AFFILIATIONS * The Skaggs Institute for Chemical Biology and Department of Chemical Physiology, The Scripps Research Institute, 10550 N.

Torrey Pines Road, La Jolla, 92037, California, USA Daniel A. Bachovchin & Benjamin F. Cravatt Authors * Daniel A. Bachovchin View author publications You can also search for this author

inPubMed Google Scholar * Benjamin F. Cravatt View author publications You can also search for this author inPubMed Google Scholar CORRESPONDING AUTHOR Correspondence to Benjamin F.

Cravatt. ETHICS DECLARATIONS COMPETING INTERESTS Benjamin F. Cravatt is a co-founder and advisor for a biotechnology company interested in developing inhibitors for serine hydrolase as

therapeutic targets. SUPPLEMENTARY INFORMATION SUPPLEMENTARY INFORMATION S1 (TABLE) Drugs that target viral and bacterial serine hydrolases. (PDF 283 kb) SUPPLEMENTARY INFORMATION S2 (TABLE)

The human serine hydrolases. (PDF 188 kb) RELATED LINKS RELATED LINKS FURTHER INFORMATION The Cravatt Laboratory GLOSSARY * Warheads Reactive chemical groups that covalently bind to

specific amino acid residues of the target enzyme. * Zymogens Inactive enzyme precursors, or pro-enzymes, that require a biochemical event (for example, a hydrolysis reaction) to convert

them into active enzymes. * Thrombi Aggregations of platelets, fibrin and cells. * Coagulation cascade A stepwise process involving the sequential activation of several serine protease

zymogens by limited proteolysis that results in the formation of fibrin blood clots. * Prodrug A pharmacological entity administered in a largely inactive form that is metabolized _in vivo_

into an active drug. * Cachexia A wasting syndrome characterized by the uncontrolled loss of muscle and adipose tissue. * Fluorescence polarization A measure of the apparent size of a

fluorophore; it is widely used to study molecular interactions. Briefly, a fluorophore excited with plane-polarized light will emit polarized light parallel to the plane of excitation unless

it rotates in the excited state. As the speed of rotational diffusion is inversely proportional to molecular volume, the resulting extent of depolarization gives a relative estimate of the

size of the fluorophore. RIGHTS AND PERMISSIONS Reprints and permissions ABOUT THIS ARTICLE CITE THIS ARTICLE Bachovchin, D., Cravatt, B. The pharmacological landscape and therapeutic

potential of serine hydrolases. _Nat Rev Drug Discov_ 11, 52–68 (2012). https://doi.org/10.1038/nrd3620 Download citation * Published: 03 January 2012 * Issue Date: January 2012 * DOI:

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