Identifying CXCR4 Receptor Agonists to Improve Diabetic Healing

鉴定 CXCR4 受体激动剂以改善糖尿病愈合

• 批准号: 9294130
• 负责人: KENNETH W LIECHTY
• 金额: $ 44.11万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-06-15 至 2019-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9294130
• 关键词: Agonist; BLR1 gene; Basic Science; Binding; Biological Assay; Biomechanics; CCR6 gene; CD69 antigen; CXCR4 Receptors; CXCR4 gene; CXCR6 gene; Catalogs; Cells; Chemicals; Chemotaxis; Clinical; Collagen; Collection; Complications of Diabetes Mellitus; Cyclic AMP; Development; Diabetes Mellitus; Diabetic Foot Ulcer; Diabetic wound; Dose; Esthesia; Expenditure; Extracellular Matrix; Fibroblasts; Fluorescence Resonance Energy Transfer; Funding; Gene Expression; Goals; Granulation Tissue; Growth Factor; Health Care Costs; Healthcare; Hospitalization; Human; Impaired wound healing; Impairment; Inflammation; Injury; Lead; Libraries; Lower Extremity; Maintenance; Mediating; Metabolic; MicroRNAs; Migration Assay; Mus; Peripheral Nervous System Diseases; Pharmaceutical Chemistry; Predisposition; Production; Property; Protocols documentation; Recruitment Activity; Reporting; Research; Signal Pathway; Skin; Skin wound; Stem cells; Technology; Testing; Time; Topical application; United States National Institutes of Health; Vascular System; Wound Healing; angiogenesis; assay development; base; chemokine; counterscreen; diabetic; diabetic wound healing; healing; improved; innovation; lentiviral-mediated; migration; novel; overexpression; prevent; receptor; receptor binding; response; scaffold; screening; small molecule; small molecule therapeutics; trafficking; wound

Summary: Impaired wound healing following injury in diabetics represents a major clinical problem, resulting in prolonged hospitalizations and significant healthcare costs. Two-thirds of all non-traumatic amputations are preceded by a diabetic wound. The development of diabetic peripheral neuropathy increases the susceptibility of diabetic skin to injury. We have shown that diabetic skin in mice and humans also has impaired skin integrity at baseline which further predisposes diabetic skin to injury. Diabetic wounds are deficient in stromal derived factor-1(SDF-1), a potent chemokine involved in progenitor cell recruitment, angiogenesis, and granulation tissue formation, mediated through binding to the CXCR4 receptor and the establishment of a chemotactic gradient. We have shown overexpression of SDF-1 corrects the diabetic wound healing impairment, and in exciting preliminary studies, can restore the integrity of diabetic skin. Given this important clinical problem, the objective of this proposal is to develop a small molecule therapeutic to target the SDF-1 receptor CXCR4 to improve diabetic skin integrity to prevent injury, and to improve wound healing should injury occur. We have developed and optimized an innovative screening approach to identify new classes of selective CXCR4 receptor agonists that can be applied topically and penetrate the skin to improve skin integrity and wound healing. We propose to screen the NIH SMR library and carry out hit-to-lead studies to identify novel CXCR4 receptor agonists that can be developed for these complications of diabetes. Aim 1. We will screen the NIH compound collection to identify molecules that can agonize the cAMP signaling pathway via the CXCR4 receptor. We have developed and optimized the primary assay to screen for first in class CXCR4 small molecule agonists and expanded our pilot screen to include 6400 compounds from our internal collection to justify screening the entire NIH SMR library using our robust testing funnel. Aim 2. We will implement dose response studies, validate all “Hits” using a counter-screen and secondary chemotaxis/migration assays, confirm direct receptor binding, and optimize our most promising molecules using SAR by catalog and medicinal chemistry approaches. Hits from Aim 1 will be counter-screened for selectivity against the related CXCR5, CCR6, CXCR6 receptors and unrelated APJ receptors. Functional assays using human cells will assess chemotaxis and migration. Cell-based binding studies will confirm direct binding to receptor, binding parameters, and optimize promising candidates by SAR. Aim 3. We will examine the ability of validated target molecules from Aim 2 to correct the abnormal expression of microRNAs that regulate inflammation, angiogenesis, and collagen synthesis in human diabetic fibroblasts, and whose expression is corrected by SDF-1. Target molecules from Aim 2 will be screened for their ability to correct expression of microRNA-146a, 15b, and 29a, which are dysregulated in diabetes, and corrected with SDF-1 treatment.

摘要：糖尿病患者受伤后伤口愈合受损是一个主要的临床问题，导致 长期住院和巨额医疗费用。所有非创伤性截肢的三分之二是 先有糖尿病伤口。糖尿病周围神经病变的发展增加了易感性 糖尿病皮肤损伤。我们已经证明，小鼠和人类的糖尿病皮肤也会损害皮肤完整性 在基线时，这进一步使糖尿病皮肤容易受伤。糖尿病伤口缺乏基质衍生的 因子 1 (SDF-1)，一种参与祖细胞募集、血管生成和肉芽形成的有效趋化因子 通过与 CXCR4 受体结合并建立趋化性介导的组织形成 坡度。我们已经证明 SDF-1α 的过度表达可以纠正糖尿病伤口愈合障碍，并且 令人兴奋的初步研究表明，可以恢复糖尿病皮肤的完整性。鉴于这一重要的临床问题， 该提案的目的是开发一种小分子疗法，以 SDF-1α 受体 CXCR4 为靶点， 改善糖尿病患者皮肤的完整性以防止受伤，并在受伤时改善伤口愈合。我们有 开发并优化了一种创新筛选方法来识别新类别的选择性 CXCR4 受体激动剂，可局部应用并渗透皮肤以改善皮肤完整性和伤口 康复。我们建议筛选 NIH SMR 文库并进行从命中到先导的研究，以鉴定新型 CXCR4 可以针对糖尿病的这些并发症开发受体激动剂。 目标 1. 我们将筛选 NIH 化合物集合，以确定可以激动 cAMP 的分子 通过 CXCR4 受体的信号通路。我们开发并优化了主要筛选方法 寻找一流的 CXCR4 小分子激动剂，并将我们的试点筛选范围扩大到包括 6400 种化合物 从我们的内部收集中证明使用我们强大的测试漏斗筛选整个 NIH SMR 库的合理性。 目标 2. 我们将实施剂量反应研究，使用反筛选验证所有“命中”，并 二次趋化/迁移测定，确认直接受体结合，并优化我们的最 通过目录和药物化学方法使用 SAR 的有前途的分子。目标 1 的命中数将是 反筛选针对相关 CXCR5、CCR6、CXCR6 受体和不相关 APJ 的选择性 受体。使用人体细胞的功能测定将评估趋化性和迁移性。基于细胞的结合 研究将确认与受体的直接结合、结合参数，并通过 SAR 优化有前途的候选药物。 目标 3. 我们将检查目标 2 中经过验证的目标分子纠正异常的能力 调节人类炎症、血管生成和胶原蛋白合成的 microRNA 的表达 糖尿病成纤维细胞，其表达受到 SDF-1α 的校正。 Aim 2 的目标分子将是 筛选了它们纠正 microRNA-146a、15b 和 29a 表达的能力，这些表达在 糖尿病，并通过 SDF-1 治疗进行纠正。