title: Buried Hydrogen Bond Interactions Contribute to the High Potency of Complement Factor D Inhibitors.
abstract: Aberrant activation of the complement system is associated with diseases, including paroxysmal nocturnal hemoglobinuria and age-related macular degeneration. Complement factor D is the rate-limiting enzyme for activating the alternative pathway in the complement system. Recent development led to a class of potent amide containing pyrrolidine derived factor D inhibitors. Here, we used biochemical enzymatic and biolayer interferometry assays to demonstrate that the amide group improves the inhibitor potency by more than 80-fold. Our crystal structures revealed buried hydrogen bond interactions are important. Molecular orbital analysis from quantum chemistry calculations dissects the chemical groups participating in these interactions. Free energy calculation supports the differential contributions of the amide group to the binding affinities of these inhibitors. Cell-based hemolysis assay confirmed these compounds inhibit factor D mediated complement activation via the alternative pathway. Our study highlights the important interactions contributing to the high potency of factor D inhibitors reported recently.
authors.affiliation: Departments of Internal Medicine and Biological Chemistry and Life Sciences Institute, University of Michigan , Ann Arbor, Michigan 48109, United States.
authors.forename: Chao-Yie
authors.initials: CY
authors.lastname: Yang
authors.affiliation: Department of Translational Hematology and Oncology Research, Taussig Cancer Institute , Cleveland Clinic, Cleveland, Ohio 44195, United States.
authors.forename: James G
authors.initials: JG
authors.lastname: Phillips
authors.affiliation: Departments of Internal Medicine and Biological Chemistry and Life Sciences Institute, University of Michigan, Ann Arbor, Michigan 48109, United States; Departments of Internal Medicine and Biological Chemistry and Life Sciences Institute, University of Michigan, Ann Arbor, Michigan 48109, United States.
authors.forename: Jeanne A
authors.initials: JA
authors.lastname: Stuckey
authors.affiliation: Departments of Internal Medicine and Biological Chemistry and Life Sciences Institute, University of Michigan , Ann Arbor, Michigan 48109, United States.
authors.forename: Longchuan
authors.initials: L
authors.lastname: Bai
authors.affiliation: Jiansu Key Laboratory of Drug Design and Optimization, Department of Medicinal Chemistry, School of Pharmacy, China Pharmaceutical University , Nanjing 210009, China.
authors.forename: Haiying
authors.initials: H
authors.lastname: Sun
authors.affiliation: Departments of Internal Medicine and Biological Chemistry and Life Sciences Institute, University of Michigan , Ann Arbor, Michigan 48109, United States.
authors.forename: James
authors.initials: J
authors.lastname: Delproposto
authors.affiliation: Departments of Internal Medicine and Biological Chemistry and Life Sciences Institute, University of Michigan , Ann Arbor, Michigan 48109, United States.
authors.forename: William Clay
authors.initials: WC
authors.lastname: Brown
authors.affiliation: Departments of Internal Medicine and Biological Chemistry and Life Sciences Institute, University of Michigan , Ann Arbor, Michigan 48109, United States.
authors.forename: Krishnapriya
authors.initials: K
authors.lastname: Chinnaswamy
keywords: Paroxysmal nocturnal hemoglobinuria; age-related macular degeneration; biolayer interferometry assay; complement factor D inhibitors; complement system; computational docking; crystal structure; enzyme inhibition assay; hemolysis assay; molecular dynamics simulations; quantum chemistry calculations; serine protease; thermodynamic integration method