Phase II Metabolite-Inspired Tripeptide Editing Generates Anti-Inflammatory Candidates for Treatment-Refractory Ulcerative Colitis
4 affiliations
- 1Digestive Disease Research Group, Institute for Biomedical Sciences, Georgia State University, Atlanta, GA 30303, USA
- 2School of Medicine, Wake Forest University, Winston-Salem, NC 27101, USA
- 3Division of Digestive Diseases, Emory University School of Medicine, Atlanta, GA 30307, USA
- 4Atlanta Veterans Affairs Medical Center, Decatur, GA 30302, USA
Abstract
Ulcerative colitis remains a major clinical challenge due to limited availability of safe, orally active anti-inflammatory drugs. Although phase II metabolism is traditionally viewed as a detoxification process, certain metabolites retain or exhibit enhanced pharmacological activity, offering an underexplored drug discovery opportunity. Here, we investigated phase II metabolite-inspired tripeptide editing using M13, a glutathione-conjugated metabolite of the anti-inflammatory natural product 6-shogaol, as a lead scaffold. A focused library of 38 analogs was generated by systematic tripeptide editing while preserving the central cysteine-containing 6-shogaol-derived scaffold. Phenotypic nuclear factor-κB (NF-κB) reporter screening identified 29 of 38 analogs with greater inhibitory activity than M13 at the screening concentration. Two selected leads, MLY2 and MLY8, demonstrated enhanced anti-inflammatory activity in Dextran Sulfate Sodium-induced experimental colitis, including restoration of colon length and reductions in fecal lipocalin-2 and pro-inflammatory cytokines. Both compounds also suppressed inflammatory cytokine production more effectively than M13 in ex vivo colonic biopsies from patients with treatment-refractory ulcerative colitis. In parallel, MLY2 and MLY8 showed no detectable mutagenicity under the Ames assay conditions and were well tolerated in a preliminary single-dose maximum tolerated dose study. Collectively, these findings establish phase II metabolite-inspired tripeptide editing as a productive lead-discovery strategy and identify MLY2 and MLY8 as anti-inflammatory leads warranting further preclinical characterization.
Related
- Exploring the therapeutic potential of Erycibe paniculata (Roxb.): metabolite profiling and antimalarial, anticancer, and modulation of NF-κB signaling pathway for anti-inflammatory activities
- Upadacitinib as a rescue therapy for steroid- and infliximab-refractory acute severe ulcerative colitis: short- and long-term effectiveness and safety in a multicenter cohort study
- Epithelial inflammatory memory in ulcerative colitis: molecular mechanisms and translational implications
- Comparative efficacy and safety of subcutaneous vedolizumab versus other targeted inflammatory bowel therapies in patients with moderate to severe ulcerative colitis: a network meta-analysis update.
- Longitudinal multi-omics profiling of tofacitinib response identifies a pre-existing Janus kinase-tractable inflammatory state in ulcerative colitis
BibTeX
@article{shi2026phase,
title = {{Phase II Metabolite-Inspired Tripeptide Editing Generates Anti-Inflammatory Candidates for Treatment-Refractory Ulcerative Colitis}},
author = {Xiaodi Shi and Rabeya Jafrin Mow and Dingpei Long and Olivier Merlin-Zhang and Emma Xu and Xiaomeng Shi and Pallavi Garg and Shanthi Srinivasan and Chunhua Yang},
journal = {International Journal of Molecular Sciences},
year = {2026},
month = sep,
doi = {10.3390/ijms27188285},
url = {https://www.mdpi.com/1422-0067/27/18/8285},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - Phase II Metabolite-Inspired Tripeptide Editing Generates Anti-Inflammatory Candidates for Treatment-Refractory Ulcerative Colitis
AU - Xiaodi Shi
AU - Rabeya Jafrin Mow
AU - Dingpei Long
AU - Olivier Merlin-Zhang
AU - Emma Xu
AU - Xiaomeng Shi
AU - Pallavi Garg
AU - Shanthi Srinivasan
AU - Chunhua Yang
PY - 2026
DA - 2026/09/17
JO - International Journal of Molecular Sciences
DO - 10.3390/ijms27188285
UR - https://www.mdpi.com/1422-0067/27/18/8285
AB - Ulcerative colitis remains a major clinical challenge due to limited availability of safe, orally active anti-inflammatory drugs. Although phase II metabolism is traditionally viewed as a detoxification process, certain metabolites retain or exhibit enhanced pharmacological activity, offering an underexplored drug discovery opportunity. Here, we investigated phase II metabolite-inspired tripeptide editing using M13, a glutathione-conjugated metabolite of the anti-inflammatory natural product 6-shogaol, as a lead scaffold. A focused library of 38 analogs was generated by systematic tripeptide editing while preserving the central cysteine-containing 6-shogaol-derived scaffold. Phenotypic nuclear factor-κB (NF-κB) reporter screening identified 29 of 38 analogs with greater inhibitory activity than M13 at the screening concentration. Two selected leads, MLY2 and MLY8, demonstrated enhanced anti-inflammatory activity in Dextran Sulfate Sodium-induced experimental colitis, including restoration of colon length and reductions in fecal lipocalin-2 and pro-inflammatory cytokines. Both compounds also suppressed inflammatory cytokine production more effectively than M13 in ex vivo colonic biopsies from patients with treatment-refractory ulcerative colitis. In parallel, MLY2 and MLY8 showed no detectable mutagenicity under the Ames assay conditions and were well tolerated in a preliminary single-dose maximum tolerated dose study. Collectively, these findings establish phase II metabolite-inspired tripeptide editing as a productive lead-discovery strategy and identify MLY2 and MLY8 as anti-inflammatory leads warranting further preclinical characterization.
SN - 1422-0067
ER -