Validation of the joint-homing and drug delivery attributes of novel peptides in a mouse arthritis model

在小鼠关节炎模型中验证新型肽的关节归巢和药物递送特性

• 批准号: 10589192
• 负责人: KAMAL D MOUDGIL
• 金额: --
• 依托单位: BALTIMORE VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-01-01 至 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10589192
• 关键词: Abbreviations; Address; Adjuvant Arthritis; Adverse effects; Adverse reactions; Affect; Aging; American; Amino Acids; Animal Model; Antibodies; Arthritis; Binding; Biodistribution; Bioinformatics; Biological; Blood Vessels; CCL2 gene; CCL3 gene; Characteristics; Chronic; Collagen Arthritis; Collagen Type II; DBA/1 Mouse; Development; Dexamethasone; Disease; Disease susceptibility; Drug Delivery Systems; Drug Targeting; Drug usage; Endothelial Cells; Evaluation; Exposure to; Financial Hardship; Genetic; HLA Antigens; Health; Health Care Costs; Healthcare; Heat shock proteins; Heterogeneity; Home; Homing; Human; IL17 gene; Immune response; Immunity; Inbreeding; Individual; Inflammation Mediators; Injections; Interleukin-1; Interleukin-6; Investigational Drugs; Joints; Laboratories; Laboratory Research; Legal patent; Ligands; Liposomes; Methodology; Methotrexate; Military Personnel; Modality; Modeling; Mus; Names; Operations Research; Oral; Oral Administration; Organ; Pain; Parents; Pathogenesis; Pathogenicity; Pathology; Patients; Peptide Phage Display Library; Peptides; Pharmaceutical Preparations; Poison; Prevalence; Quality of life; RANTES; Rattus; Rehabilitation therapy; Reporting; Research Personnel; Rheumatoid Arthritis; Risk; Stress; Surface; Susceptibility Gene; Synovitis; T-Lymphocyte; TNFRSF10A gene; Technology; Testing; Therapeutic; Therapeutic Index; Tissues; Toxic effect; Trademark; Treatment Efficacy; United States Department of Veterans Affairs; Validation; Vascular Endothelial Cell; Vascular Endothelium; Veterans; Wild Type Mouse; Woman; antiarthritic agent; arthritis therapy; arthropathies; chemokine; combat zone; comparative; comparative efficacy; cyanine; cyanine dye; cytokine; disability; drug metabolism; genetic makeup; human model; humanized mouse; immunoregulation; improved; in vivo; innovation; insight; joint inflammation; joint injury; liposomal formulation; men; migration; military veteran; molecular marker; nanoparticle; near infrared dye; neoplastic cell; novel; novel therapeutics; pharmacokinetics and pharmacodynamics; pilot test; risk/benefit ratio; screening; systemic toxicity; translation to humans; translational applications; translational study

Rheumatoid arthritis (RA) is a chronic debilitating disease affecting over 1% of Americans. Women are affected about 3 times more often than men. RA also constitutes a major health problem among the U.S. Veterans. The Veteran population is aging, and the number of women serving in the military is gradually increasing. In addition, military personnel in combat zones are exposed to extreme stress and toxic substances in different forms. All these factors may directly or indirectly influence host immunity and induction/aggravation of RA. The pain, discomfort, lost working days, and disabilities caused by RA can adversely affect Veterans’ quality of life in many ways, as well as impose heavy cost of healthcare and rehabilitation. Therefore, studies advancing new modalities of treatment of RA are highly relevant to the U.S. Veterans. Using the rat adjuvant-induced arthritis (AA) model of human RA, we developed a peptide-guided targeted liposomal drug delivery system for arthritis therapy. For this purpose, we used a novel joint-homing peptide ART-2 to guide liposomes entrapping dexamethasone (DEX) to arthritic joints of rats. Peptide-guided liposomal DEX was more effective in suppressing arthritis in rats than untargeted liposomal drug or free drug. Importantly, the increased efficacy of the ART-2 liposomal group was not associated with concurrent increase in systemic toxicity. Thus, peptide- guided liposomal drug delivery improved the therapeutic profile (benefit/risk ratio) of DEX over the other two modalities. However, unlike inbred rats (used in the rat AA model), which are genetically homogeneous (same) and generate immune response mainly to heat-shock protein 65 (Hsp65), RA patients have diverse genetic make-up and generate immune response to both Hsp65 and type II collagen (CII). Therefore, we need to validate our current findings from the rat AA model in an independent second model, the mouse collagen- induced arthritis (CIA) model, which differs in genetics and certain disease parameters from the AA model. This validation will be the first step to de-risk the translational application of this technology to RA patients before undertaking further development and evaluation to meet the criteria of an investigational new drug (IND) for the FDA. This is significant because the Veterans Affairs (VA) has filed a non-provisional patent on the use of peptide ART-2 for arthritis therapy with the U.S. Patent and Trademark Office. The AA and CIA models of arthritis share some (but not all) features with human RA, but they also differ in a few critical characteristics: 1) unlike Lewis rats (RT-1l), inbred wild type mice do not develop AA; 2) the predominant target of the T cells/antibodies is CII in CIA, but Hsp65 in AA; 3) antibodies are pathogenic in CIA, but either show no effect or are protective in AA; and 4) drug metabolism is different in rats and mice. We will use two CIA models: wild type DBA/1 (H-2q) mice and “humanized” mice (H-2b) expressing HLA-DR4, one of the disease-susceptibility genes for RA. [We will test methotrexate (MTX), the gold-standard for RA. We have successfully prepared liposomes entrapping MTX, and pilot testing in the parent rat arthritis model (AA) showed that MTX entrapped in liposomes is biologically active and effective in suppressing arthritis. Recently, we have completed setting-up of the CIA model in our laboratory to validate the above findings from AA model.] The specific aims of our study in CIA model based on peptide ART-2 and methotrexate (MTX) are: Aim 1: (a) To validate the preferential in-vivo homing of systemically administered peptide ART-2-displaying liposomes entrapping a near-infrared dye (Cyanine 7) to arthritic joints of mice with CIA; and (b) To examine the pharmacokinetic and pharmacodynamic aspects of liposomal MTX delivery in mice having CIA. Aim 2: (a) To validate the comparative efficacy of MTX delivery via peptide ART-2-displaying liposomes, control liposomes, and free drug in suppressing arthritis in CIA mice; and (b) To determine the relative multiorgan toxicity of these three drug delivery modalities to assess their comparative therapeutic efficacy/toxicity profiles. We believe that with appropriate validation, this technology can be adapted and further refined for use in Veterans with RA.

风湿性关节炎（RA）是一种慢性衰弱性疾病，影响超过1%的美国人。妇女受到影响 是男性的3倍。RA也构成美国退伍军人的一个主要健康问题。的 退伍军人人口正在老龄化，在军队服役的妇女人数正在逐渐增加。在 此外，在战区的军事人员在不同的环境中面临着极端的压力和有毒物质， forms.这些因素都可能直接或间接地影响宿主的免疫功能，从而诱发或加重RA的发生。的 类风湿关节炎引起的疼痛、不适、工作日损失和残疾会对退伍军人的生活质量产生不利影响 以及造成沉重保健和康复费用。因此，新的研究进展 RA的治疗方式与美国退伍军人高度相关。使用大鼠泻药诱导的关节炎 (AA)以人类风湿关节炎为模型，我们开发了一种肽导向的关节炎靶向脂质体给药系统 疗法为此，我们使用了一种新的联合归巢肽ART-2来引导脂质体包封 地塞米松（DEX）对大鼠关节炎关节的影响。肽引导的脂质体DEX在治疗中更有效。 抑制大鼠关节炎的作用优于非靶向脂质体药物或游离药物。重要的是， ART-2脂质体组与全身毒性同时增加无关。因此，肽- 与其他两种药物相比，引导脂质体药物递送改善了DEX的治疗特征（获益/风险比） 方式。然而，与近交系大鼠（用于大鼠AA模型）不同，近交系大鼠具有遗传同质性（相同） 并主要对热休克蛋白65（Hsp 65）产生免疫应答，RA患者具有多种遗传学特征， 补充并产生对Hsp 65和II型胶原（CII）的免疫应答。因此我们需要 验证了我们目前的发现，从大鼠AA模型在一个独立的第二个模型，小鼠胶原蛋白- 诱导性关节炎（CIA）模型，其在遗传学和某些疾病参数上与AA模型不同。 该验证将是降低该技术在RA患者中的转化应用风险的第一步 在进行进一步开发和评估以满足研究性新药（IND）的标准之前 为FDA。这是重要的，因为退伍军人事务部（VA）已经提交了一份非临时专利的使用 用于关节炎治疗的肽ART-2与美国专利和商标局。AA和CIA的模式 关节炎与人类RA有一些（但不是全部）特征，但它们也有一些关键特征不同： 1)与刘易斯大鼠（RT-1 l）不同，近交系野生型小鼠不发生AA; 2）T的主要靶点 细胞/抗体在CIA中是CII，但在AA中是Hsp 65; 3）抗体在CIA中是致病性的，但没有显示出作用， 在AA中具有保护作用; 4）大鼠和小鼠的药物代谢不同。我们将使用两个中情局模型：野生 DBA/1型（H-2 q）小鼠和表达HLA-DR 4（疾病易感性之一）的“人源化”小鼠（H-2b） RA的基因[We将测试甲氨蝶呤（MTX），RA的金标准。我们成功地准备了 脂质体包埋MTX，并在母体大鼠关节炎模型（AA）中进行初步试验，结果表明MTX 包埋在脂质体中的药物具有生物活性，并有效抑制关节炎。最近，我们完成了 在本实验室建立CIA模型以验证AA模型的上述发现。具体 本研究以ART-2肽和甲氨蝶呤（MTX）为基础建立CIA模型，目的1：（a） 验证了全身施用的肽ART-2展示脂质体的优先体内归巢 将近红外染料（花青7）包埋到患有CIA的小鼠的关节炎关节;和（B）为了检查 在患有CIA的小鼠中脂质体MTX递送的药代动力学和药效学方面。目标2：（a） 验证通过肽ART-2展示脂质体、对照脂质体 （B）测定这些化合物的相对多器官毒性， 三种药物递送模式以评估它们的比较治疗功效/毒性特征。我们认为 通过适当的验证，该技术可以适应并进一步改进用于患有RA的退伍军人。