Work Description

Date:
12 March, 2025

Dataset Title:
CYP2W1 Binding Characterization with Azole Library

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:
- The dataset presented herein estimates the binding affinity for 104 imidazole-containing compounds to cytochrome P450 2W1.

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:
The methods for data collection are detailed in the publication cited below. The assay was performed using a plate reader to determine changes in absorbance of the P450 spectra in the UV-visible range. Changes in absorbance are quantified and plotted against concentration of compound in each well. The nonlinear regression one site, specific binding in GraphPad Prism was used to determined dissociation constants and maximum changes in absorbance.
Instrument and/or Software specifications: Perkin Elmer EnVision plate reader, GraphPad Prism

Files contained here:

2W1_compiled_Kd_supplemental.xlsx
-The file deposited here contains each compound included in the azole library along with chemical properties of interest. Dissociation constants, maximum changes in absorbance, and the ratio of maximum change in absorbance to dissociation constant are reported for compounds that cause a spectral shift with cytochrome P450 2W1.

2W1_compiled_Kd_supplemental.csv
-.CSV version of the same data.

Related publication(s):
Frydendall, E. K. and E. E. Scott (2024). "Development of a high throughput cytochrome P450-ligand binding assay." J Biol Chem: 107799.

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/v4nh-hk80.