Peptide Therapeutics for Inhbiting Abdominal Adhesions

DESCRIPTION (provided by applicant): Adhesions, fibrous scars abnormally conjoining adjacent tissues, are primarily caused by surgical procedures. They can arise in many areas of the body as common sequelae of abdominal, gynecological, thoracic, and cardia c procedures [4, 5, 8-10]. Adhesions can cause not only pain and discomfort, but also loss of organ function and even death. While the technology described herein will effect prevention of all adhesion types, the high-need, high-burden abdominal (specifica lly small bowel) adhesion prevention application will be the focus of this grant submission. Burden of disease: Abdominal adhesions occur at high frequency and have tremendous associated burden of disease. Adhesions form in conjunction with virtually every abdominal surgery. Multiple studies cite that, of those patients who have abdominal surgery, up to 93% will develop adhesions [1, 2]. Transforming Growth Factor-21 and other cytokines are known to initiate adhesion formation through activation of mitogen activated protein kinase activated kinase, (MK2). MK2 is implicated in connective tissue growth factor (CTGF) and collagen type I up regulation. The proposed MK2 inhibitor peptide therapeutic will inhibit the scar inducing activities thus offering the pote ntial of being a rapid, targeted therapy to prevent adhesions [7]. Through this phase I SBIR plan final optimization and selection of the therapeutic peptide, in addition to initial purity, solubility and stability testing will be achieved. The following a ims will be competed: 1) Optimization and specificity testing of MK2 inhibitors will be completed. Based upon knowledge gained in preliminary experiments, peptide variants will be synthesized to improve activity while maintaining specificity. Activ ity will be evaluated using an in vitro kinase assay. Specificity of the four top drug candidates will be tested in a kinase profiler assay to determine if enhancing MK2 inhibition activity affects specificity. Finally, evaluation of MK2i inhibition of CTG F expression in a human fibroblast cell line will be performed. 2) Preliminary biophysical evaluation: purity, solubility, stability. Purity will be determined by elemental analysis and quantitative amino acid analysis. Maximum solubility testing w ill be performed in PBS and confirmed using quantitative amino acid analysis. To predict drug product stability, we will perform preliminary aqueous-based stability testing. Candidates will be solubilized in buffer, stored at 4, 25 or 40 C for four weeks a nd degree of degradation will be evaluated using an HPLC assay.