Molecular Pharmacy (Ricklin)

The complement system is an important innate immune pathway that plays critical roles as a first line of defense against invading organisms, damaged tissues, and the maintenance of healthy cell populations. However, its uncontrolled activation or insufficient regulation contributes to diverse clinical conditions affecting millions of patients worldwide, such as ischemia-reperfusion injury (IRI; during myocardial infarction, stroke, or organ transplantation), age-related macular degeneration, and several neurodegenerative disorders. Therefore, complement has steadily moved into the focus of drug development efforts. Whereas most therapeutic strategies focus on blocking individual effector functions, preventing the initial activation of complement could offer distinct advantages.

Collectin-11 (CL-11), a soluble C-type lectin that recognizes mannose (Man)- and fucose (Fuc)-containing glycans at the surface of pathogens or damaged cells is one such target of interest. CL-11 was recently implicated in the pathology of both acute and chronic kidney injury. In IRI-mediated acute kidney injury associated with transplantation, the expression of CL-11 is significantly elevated and co-localizes with Fuc-rich cell-surface glycans to initiate complement-dependent destruction of the kidney and loss of organ function. In chronic kidney injury, CL-11 binds to Man-based structures and acts as a leukocyte chemoattractant and enhancer of fibroblast proliferation, resulting in renal fibrosis. CL-11 also activates complement in retinal stem cells, restricting the use of stem cell transplantation for patients with age-related macular degeneration, the leading cause of blindness in the developed world. In both cell and animal models, these harmful immune responses were suppressed: (i) in CL-11 knockout mice; (ii) through enzymatic cleavage of cell-surface Man/Fuc; or (iii) by treatment with high concentrations of soluble Man/Fuc. While high-dose Fuc therapy largely prevented complement-mediated tissue damage in mice, this treatment option has real-world limitations concerning specificity, dose requirements, and pharmacokinetic (PK) profile. Therefore, the availability of CL-11-specific inhibitors with improved efficacy and drug-like properties is of therapeutic interest and would provide an important tool for delineating CL-11’s (patho)physiological role in disease.

This joint proposal between lead investigators Dr. Rachel Hevey and Prof. Martin Smieško combines their strong and synergistic expertises in computational modeling, glycomimetic drug design, chemical synthesis, and functional evaluation, to develop a novel class of specific CL-11 inhibitors. Based on the emerging role of CL-11 in renal disease and IRI, we envision that the development of carbohydrate-based, selective inhibitors for CL-11 could eventually provide a promising therapy for preventing or reducing tissue damage. To our knowledge, there are no active development programs for glycomimetic inhibitors of CL-11, and therefore the development of molecules with high affinity, selectivity, and suitable PK properties constitutes an innovative project with a potential for real-world impact.

Current estimates suggest that nearly 1.3 million lives are lost annually to antibiotic-resistant infections, with this number rising to 10 million by 2050. Antibiotic resistance results in failed treatment, prolonged hospital stays, and increases the financial burden placed on medical systems and society. Therefore, novel approaches to antibiotic use are urgently needed.

Recently, a novel class of anti-virulence compounds based on mucosal glycans were discovered, which are active against a range of pathogenic cross-kingdom species, including fungal, Gram-positive, and Gram-negative bacterial pathogens. These mucin O-glycans do not directly affect pathogen survival but instead repress virulence, thereby increasing susceptibility to host immune pathways and indirectly reducing infection load. By exerting their activity through virulence attenuation instead of broad-spectrum elimination, these compounds can promote the reestablishment of a healthy microbiome and are expected to be at a reduced risk of developing drug resistance. Although this exciting new class of compounds has demonstrated broad activity, the mechanism(s) by which these glycans act has not yet been elucidated. By identifying their discrete molecular target(s), specific binding interactions can be analyzed and more drug-like glycomimetic therapies can be developed.

Mucins contain hundreds of different O-glycans which are not commercially available, cannot be purified as single structures from native sources, and are not readily amenable to solid-phase synthesis. Therefore, in ongoing work we have been developing a platform to generate individual mucin glycan structures and evaluate their ability to regulate pathogen virulence. In the further development and application of this platform, specific aims of the current proposal are focused on: (i) the recombinant expression of predicted virulence-regulating proteins; (ii) the investigation of mucin glycan binding to virulence-regulating proteins through parallel techniques; and (iii) further expansion of the mucin glycan library and the development of next-generation glycomimetic molecules.

The proposed project will improve our understanding of mucus–pathogen relationships, provide insights into virulence-attenuating mechanism(s), and provide the next steps to development of an innovative therapeutic.