# SMALL MOLECULE BACTERIAL LECTIN ANTAGONISTS FOR UTI TREATMENT AND PREVENTION

> **NIH NIH R01** · WASHINGTON UNIVERSITY · 2020 · $549,282

## Abstract

Gram-negative bacteria utilize extracellular fibers called chaperone-usher pathway (CUP) pili to mediate adhesion
to host and environmental surfaces, facilitate invasion into host tissues, and promote interaction with other bacteria
to form biofilms. Uropathogenic E. coli (UPEC) use a CUP adhesion protein (lectin) called FimH on the type 1 pilus
to bind to mannosylated glycoproteins on bladder epithelial cells to mediate the onset and progression of urinary
tract infections (UTIs). This binding event initiates bacterial invasion and formation of intracellular bacterial 
communities (IBCs) in the eukaryotic cell. Using rational drug design, mannoside antagonists of FimH have been developed as orally bioavailable therapeutics for the treatment and prevention of UTIs. Acute infection can either 
self-resolve or develop into chronic cystitis, which is characterized by: i) persistent, high titer bacteriuria and 
bacterial bladder burdens at sacrifice >4 weeks post- infection; ii) chronic inflammation and urothelial necrosis;
iii) lymphonodular hyperplasia in the bladder submucosa and; iv) urothelial hyperplasia with a lack of uroplakin expression, which is a marker for terminal differentiation in superficial facet cells. Similar histological findings
have been observed in humans suffering persistent bacteriuria and recurrent UTI. FmlH, the tip-associated 
adhesin of Fml/F9/Yde pili (previously denoted FmlD) functions in UPEC pathogenesis by providing a fitness
advantage during chronic cystitis. FmlH specifically binds to Gal(β1-3)GalNac epitopes which appear as part of 
a remodeled glycan/galactose profile of the mouse bladder during chronic cystitis. FmlH is also upregulated in
urines directly isolated from patients with UTI compared to expression during in vitro growth in media or normal 
urine, suggesting a host-specific induction. In this proposal, innovative strategies will be taken to rationally 
develop βGal and βGalNAc ligands as antagonists of FmlH, alone or in combination with FimH inhibitors, as
new preclinical therapeutics for treatment of acute and chronic cystitis. Using an X-ray structure of FmlH and
virtual screening, new FmlH ligands, including O-nitrophenyl-β-galactoside (ONPG) were identified. 
Subsequently, the X-ray structures of Galβ-1-3-GalNAc (TF) and ONPG bound to FmlH were used to design 
improved FmlH ligands, with binding affinities several times higher than ONPG, as determined by an ELISA 
binding assay. This structure-based design strategy will be used to optimize new ligands with higher affinity
and good drug-like properties. This project's aims will be accomplished by integrating: i) X-ray structure-based drug design with medicinal chemistry (Aim 1); ii) biochemical screening, functional cell and tissue binding assays 
Aim 2) and; rigorous pharmacokinetic (PK) evaluation and pharmacodynamic (PD) efficacy testing of the most
promising FmlH ligands, in murine animal models of chronic cystitis, urosepsis/kidney infection, ...

## Key facts

- **NIH application ID:** 9872163
- **Project number:** 5R01DK108840-04
- **Recipient organization:** WASHINGTON UNIVERSITY
- **Principal Investigator:** SCOTT J. HULTGREN
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $549,282
- **Award type:** 5
- **Project period:** 2017-02-01 → 2021-07-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9872163

## Citation

> US National Institutes of Health, RePORTER application 9872163, SMALL MOLECULE BACTERIAL LECTIN ANTAGONISTS FOR UTI TREATMENT AND PREVENTION (5R01DK108840-04). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/9872163. Licensed CC0.

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