New Peptides for the Treatment of Multiple Sclerosis

治疗多发性硬化症的新肽

• 批准号: 8473290
• 负责人: Christine Beeton
• 金额: $ 35.12万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-01 至 2016-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8473290
• 关键词: Acute; Adverse effects; Affect; Animal Model; Arrhythmia; Arthritis; Binding Sites; Biological; Carbon; Cardiotoxicity; Cell Enlargement; Cells; Central Nervous System Diseases; Cessation of life; Charge; Chlamydia; Chlamydia trachomatis; Chronic; Coupled; Data; Delayed Hypersensitivity; Development; Disease; Docking; Drug Formulations; Drug Kinetics; Experimental Autoimmune Encephalomyelitis; FDA approved; Generations; Goals; Half-Life; Human; Hydrolysis; Immune response; Immunomodulators; Immunosuppression; Immunotherapy; In Vitro; Infection; Inflammatory; Influenza; Injection of therapeutic agent; Intervention; Ion Channel; Kv1.2' channel; Lymphocyte Activation; Malignant Neoplasms; Modeling; Modification; Molecular Models; Multiple Sclerosis; Mus; Pathogenesis; Peptides; Pharmaceutical Preparations; Play; Polyethylene Glycols; Potassium Channel; Pristane; Production; Proteins; Public Health; Rattus; Relapse; Relapsing-Remitting Multiple Sclerosis; Resolution; Role; Safety; Sea Anemones; Seizures; Site; Specificity; Structure; T memory cell; T-Lymphocyte; T-Lymphocyte Subsets; Temperature; Testing; Therapeutic Intervention; Tissues; Toxic effect; United States; Viral; Voltage-Gated Potassium Channel; adduct; analog; base; cell motility; channel blockers; compliance behavior; cost; cytokine; design; disability; high risk; immunogenic; immunogenicity; improved; in vitro testing; in vivo; meetings; molecular dynamics; molecular modeling; nonhuman primate; novel; novel therapeutics; oxidation; patch clamp; pathogen; polypeptide; safety study; terminally differentiated effector memory (TEM) T cells; tool

DESCRIPTION (provided by applicant): Multiple sclerosis (MS) is a chronic inflammatory disease of the central nervous system that leads to severe disability and death. Of the current FDA-approved medications for relapsing forms of MS, all have significant side effects. We propose to use a multi-disciplinary, structure-based approach to developing novel polypeptide blockers of the voltage-gated potassium channel Kv1.3 as new interventions for MS. This channel is highly expressed by terminally-differentiated effector memory T (TEM) lymphocytes that play a major role in MS pathogenesis. Extensive in vitro and in vivo efficacy and safety studies have validated Kv1.3 as a target for immunotherapy and shown that the peptide ShK, originally isolated from a sea anemone, is not only a potent blocker of this channel but also an effective immunomodulator. However, its lack of selectivity for Kv1.3 channels creates a high risk of toxicity through interactions with other potassium channels. We have developed a first generation of synthetic analogs of ShK by modifying its N-terminus with non-protein adducts. These analogs show enhanced specificity for Kv1.3 over other ion channels while retaining picomolar potency, and they selectively suppress cytokine production and proliferation of human TEM cells without affecting other T cell subsets. In rat studies, one of these analogs (ShK-186) suppresses TEM cell enlargement and motility in inflamed tissues, inhibits delayed-type hypersensitivity, and effectively treats chronic-relapsing experimental autoimmune encephalomyelitis (CR-EAE; a model of MS) and pristane-induced arthritis. While ShK-186 and related analogs have an excellent safety profile in rats and do not compromise the protective immune response to acute infection with viral (influenza) or bacterial (chlamydia) pathogens, they suffer from several limitations: (i) they are sensitive to changes in pH and temperature; (ii) they have very short in vivo half-lives; (iii) a phosphorylated residue on ShK-186 can be dephosphorylated; (iv) they contain a Met residue that is susceptible to oxidation; and (v) their non-protein adducts are immunogenic. We now propose to design, generate, and evaluate novel analogs of ShK. Under Specific Aim 1 we will use molecular modeling and high-resolution NMR to determine the docking configuration of ShK analogs on Kv1.3 and thereby to design and synthesize more potent and selective N- and C-terminally extended ShK analogs. Under Specific Aim 2 we will assess ShK analogs for their in vitro potency, selectivity, stability, and effects on T lymphocyte activation, and PEGylate them to increase their circulating half-life. Under Specific Aim 3 we will evaluate the most potent and selective ShK analogs in vivo for pharmacokinetics, immunogenicity, safety, and efficacy. This project will generate novel peptide blockers of Kv1.3, which we believe will be valuable leads in the development of new treatments for MS and other chronic inflammatory diseases.

描述（由申请人提供）：多发性硬化症（MS）是一种中枢神经系统慢性炎症性疾病，可导致严重残疾和死亡。目前 FDA 批准的治疗复发型多发性硬化症的药物均具有显着的副作用。我们建议使用多学科、基于结构的方法来开发电压门控钾通道 Kv1.3 的新型多肽阻断剂，作为 MS 的新干预措施。该通道由终末分化效应记忆 T (TEM) 淋巴细胞高度表达，在 MS 发病机制中发挥重要作用。广泛的体外和体内功效和安全性研究已验证 Kv1.3 作为免疫治疗的靶点，并表明最初从海葵中分离出来的肽 ShK 不仅是该通道的有效阻断剂，而且还是一种有效的免疫调节剂。然而，它对 Kv1.3 通道缺乏选择性，通过与其他钾通道相互作用而产生高毒性风险。我们通过用非蛋白质加合物修饰 ShK 的 N 末端，开发出了第一代 ShK 的合成类似物。这些类似物表现出比其他离子通道更强的 Kv1.3 特异性，同时保留皮摩尔效力，并且它们选择性抑制人 TEM 细胞的细胞因子产生和增殖，而不影响其他 T 细胞亚群。在大鼠研究中，其中一种类似物 (ShK-186) 可抑制发炎组织中 TEM 细胞的增大和运动，抑制迟发型超敏反应，并有效治疗慢性复发性实验性自身免疫性脑脊髓炎（CR-EAE；MS 模型）和降植烷诱发的关节炎。虽然 ShK-186 和相关类似物在大鼠中具有出色的安全性，并且不会损害对病毒（流感）或细菌（衣原体）病原体急性感染的保护性免疫反应，但它们存在一些局限性：（i）它们对 pH 和温度的变化敏感； (ii) 它们的体内半衰期非常短； (iii)ShK-186上的磷酸化残基可以去磷酸化； (iv) 它们含有易被氧化的甲硫氨酸残留物； (v)它们的非蛋白质加合物具有免疫原性。我们现在建议设计、生成和评估 ShK 的新型类似物。在具体目标 1 下，我们将使用分子建模和高分辨率 NMR 来确定 ShK 类似物在 Kv1.3 上的对接构型，从而设计和合成更有效和选择性的 N 端和 C 端延伸的 ShK 类似物。在具体目标 2 下，我们将评估 ShK 类似物的体外效力、选择性、稳定性以及对 T 淋巴细胞活化的影响，并对它们进行聚乙二醇化以延长其循环半衰期。在具体目标 3 下，我们将评估体内最有效和选择性最强的 ShK 类似物的药代动力学、免疫原性、安全性和功效。该项目将产生新型 Kv1.3 肽阻断剂，我们相信这将在开发多发性硬化症和其他慢性炎症性疾病新疗法方面发挥重要作用。