Work Description

Date:
12 March, 2025

Dataset Title:
Library of Compounds Tested for CYP2W1 Inhibition

Dataset Contact:
Emily Scott [email protected]

Dataset Creators:

Name: Elyse K. Frydendall
Institution: University of Michigan, Ann Arbor
Email: [email protected]
ORCID: https://orcid.org/0000-0002-1862-0529

Name: Emily E. Scott
Institution: University of Michigan, Ann Arbor
Email: [email protected]
ORCID: https://orcid.org/0000-0002-5538-5716

Funding:
Pharmacological Sciences Training Program (PSTP) T32GM007767, R37 GM076343 (NIH)

Key Points:
-Of the 5957 compounds tested for CYP2W1 inhibition, 694 were initially identified as inhibitors.

Research Overview:
Cytochrome P450 2W1 is an orphan P450 enzyme with little known about its endogenous ligands or function. CYP2W1 protein expression has been identified in fetal colon tissue before 18 weeks of gestation and in several cancers but is silenced in healthy adult human tissues. Although the known ligand profile for CYP2W1 is limited, it has been identified as an activator of certain duocarmycin prodrugs in vitro and in vivo for the treatment of colorectal cancers that express CYP2W1 protein. However, CYP1A1 can also activate these prodrugs and is expressed in healthy adult tissue, putting such tissues at risk of the ultrapotent cytotoxic effects of duocarmycins. Therefore, this study aims to identify structural features in the CYP2W1 active site that may be exploited to design a duocarmycin prodrug selectively activated by CYP2W1. In lieu of a protein structure, the CYP2W1 ligand profile was investigated using multiple screens. In total, 694 inhibitors of CYP2W1 metabolism were identified. Furthermore, a more focused azole library identified 67 compounds that coordinate to the CYP2W1 active site heme iron, meaning they are inhibitors. These 67 azoles were overlaid to highlight key features of ligands that interact with CYP2W1 via pharmacophore models. The pharmacophores revealed one hydrophobic and two aromatic ring features consistent among compounds binding with high and low affinity. However, a hydrogen bond acceptor was identified that is positioned in front of or behind the plane of the azole ring depending on ligand affinity. Further investigation into the physiochemical features of the CYP2W1 active site may inform the development of selective duocarmycin prodrugs.

Methodology:
Inhibitors were evaluated based on their ability to inhibit CYP2W1 O-demethylation of Luciferin-1A2. When cysteine is introduced to the product of this reaction, luciferin is formed. Luciferin formation is quantified based on luminescence. Data were collected as raw luminescence values. Inhibitors that resulted in luminescence values that exceeded three times the standard deviation of controls containing no inhibitor were identified as hits. 

Instrument and/or Software specifications:
Perkin Elmer EnVision plate reader, GraphPad Prism

Files contained here:

CYP2W1_high_throughput_inhibition_library.xlsx
-This table contains the names, molecular mass (g/mol), and SMILES structure for all compounds tested for CYP2W1 inhibition in the high throughput screen described in this chapter in the tab titled “Compound Library”. Compounds that were identified as CYP2W1 inhibitors are listed in the tab titled “CYP2W1 inhibitors.” This tab contains compound names, molecular mass (g/mol), average standard deviation, number of experiments in which a compound was classified as a hit, number of experimental replicates, hit rate, and SMILES structure.

CYP2W1_high_throughput_inhibition_library_CompoundLibrary.csv
CYP2W1_high_throughput_inhibition_library_Inhibitors.csv
-.CSV version of the same data.

Use and Access:
This data set is made available under an Attribution-NonCommercial 4.0 International license (CC BY-NC 4.0).

To Cite Data:
Frydendall, E. (2025). Human cytochrome P450 enzymes in drug and fatty acid metabolism [Doctoral dissertation, University of Michigan]. University of Michigan Library Deep Blue Data. https://doi.org/10.7302/81h9-qk39 .