Novel Glycosaminoglycan Derivatives for Treatment of Bladder Inflammation

描述（由申请人提供）：膀胱疼痛综合征/间质性膀胱炎（PBS/IC）是一种惰性膀胱疾病，一直是一种使人衰弱的疾病，几乎没有真正有效的治疗选择。影响膀胱的炎症状况可导致盆腔疼痛、衰弱的泌尿系统症状、膀胱纤维化、复发性泌尿系统感染和肾衰竭。我们已经建立了一个小鼠膀胱炎症模型，从酒渣鼻皮肤病变的病理生理推断，表达高水平的抗菌抗菌肽LL-37。这种高度阳离子化的防御素与酒渣鼻的炎症反应相关，并引起炎症反应。重要的是，LL-37在泌尿系统中自然产生，在尿路感染发作期间显着上调，并似乎引发严重的膀胱炎症。目前有两种抗炎硫酸多糖用于治疗，但都不是特别有效。首先，肝素是静脉注射，但抗凝血特性和费用限制了它的常规使用。其次，Elmiron（聚硫酸戊聚糖）是口服给药，起效时间较长，仅对<50%的女性有效，生物利用度差。无硫糖胺聚糖（GAG），透明质酸（HA），在美国以外的地方作为Cystistat可用，但疗效较低。需要更好的治疗方法。在这个I期项目中，GlycoMira将测试使用新型抗炎GAG衍生物减轻ll -37介导的膀胱炎症的可行性。GlycoMira正在开发一种新型的非抗凝血，抗炎硫酸多糖，作为安全有效的炎症调节剂，半合成糖胺聚糖醚（SAGEs）。sage还抑制了许多加剧炎症的途径，包括P-和l -选择素结合、阳离子蛋白酶活性和晚期糖基化终产物（RAGE）受体的激活。具体来说，在这个I期SBIR项目中，GlycoMira将在两个特定目的中探索使用SAGE GM-1111包覆膀胱尿上皮和减少ll -37介导的膀胱炎症的可行性。首先，通过膀胱内滴注一种荧光生物偶联物AlexaFluor-GM-1111来检测sage对膀胱组织的定位、结合和渗透，并与alexafluor标记的肝素和HA进行比较。其次，我们将在膀胱内灌注LL-37的膀胱炎症模型中，通过GM-1111、肝素或HA预处理或后处理来测试GM-1111的治疗潜力。组织将进行组织学分析髓过氧化物酶活性，以量化中性粒细胞浸润。初步数据表明GM-1111包覆尿上皮并减少膀胱炎症。GlycoMira在犹他大学的合作泌尿科医生认识到GM-1111在膀胱炎症临床治疗中的潜力。

DESCRIPTION (provided by applicant): Painful bladder syndrome/interstitial cystitis (PBS/IC) is an indolent bladder disorder that has continued to be a debilitating disease with few truly effective treatment options. Inflammatory conditions that afflict the urinary bladder can lead to pelvic pain, debilitating urinary symptoms, bladder fibrosis, recurring urinary infection, and renal failure. We have developed a murine model of bladder inflammation extrapolated from the pathophysiology of skin lesions in rosacea, which express high levels of the antimicrobial cathelicidin peptide LL-37. This highly cationic defensin is both correlated with, and causative for, the profound inflammatory responses in rosacea. Importantly, LL-37 is naturally produced in the urinary system, is significantly upregulated during urinary tract infection episodes, and appears to trigger profound bladder inflammation. Two anti-inflammatory sulfated polysaccharides are currently used for therapy, but neither is particularly effective. First, heparin is administered intravesically, but the anti-coagulant properties and expense limit its regular usage. Second, Elmiron (pentosan polysulfate) is administered orally, has a long lead time for onset of efficacy, is only effective in <50% of women, and is poorly bioavailable. The unsulfated glycosaminoglycan (GAG), hyaluronan (HA), is available outside the US as Cystistat but has low efficacy. A better treatment is needed. In this Phase I project, GlycoMira will test the feasibility of using novel anti-inflammatory GAG derivatives to mitigate LL-37-mediated bladder inflammation. GlycoMira is developing a new class of non-anticoagulant, anti-inflammatory sulfated polysaccharides as safe and effective inflammation-modulating agents, the semi-synthetic glycosaminoglycan ethers (SAGEs). SAGEs also inhibit numerous pathways that exacerbate inflammation, including P- and L-selectin binding, cationic protease activity, and activation of the receptor for advanced glycation end-products (RAGE). Specifically, in this Phase I SBIR project, GlycoMira will explore the feasibility of using the SAGE GM-1111 to coat bladder uroepithelium and to reduce LL-37-mediated bladder inflammation in two Specific Aims. First, the localization, binding, and penetration of the bladder tissues by SAGEs will be examined by intravesical instillation of a fluorescent bioconjugate, AlexaFluor-GM-1111 and compared with AlexaFluor-labeled heparin and HA. Second, we will test the therapeutic potential of GM-1111 by pre-treatment or post-treatment with GM-1111, heparin, or HA in the model of bladder inflammation by intravesical instillation of LL-37. Tissues will be analyzed histologically for myeloperoxidase activity to quantify neutrophil infiltration. Preliminary data suggest that GM-1111 coats the uroepithelium and reduces bladder inflammation. GlycoMira's collaborating urologists at the University of Utah recognize the potential of GM-1111 for clinical treatment of bladder inflammation. PUBLIC HEALTH RELEVANCE: Inflammatory conditions that afflict the urinary bladder are a significant urologic health concern to many in the United States. Specifically in women afflicted with a debilitating condition known as painful bladder syndrome/interstitial cystitis (PBS/IC), these inflammatory processes can lead to severe symptoms characterized by urinary frequency, bladder pain, nocturia, urgency, and pelvic pain. The proposed studies are innovative and aim to understand the cause of bladder inflammation and to develop a new treatment. A physiologically relevant method to create bladder inflammation will be developed to unravel novel pathways that perpetuate the disease process. In addition, novel therapeutic sulfated polysaccharides will be examined for their efficacy in successfully treating inflammation in this model. The ultimate goal of this proposal is to both gain a better understanding of bladder inflammatory pathogenesis, and to provide a safe and effective new treatment for the many patients who suffer from PBS/IC.