Silk Fibroin Small-Molecule Drug Carriers for the Treatment of Osteoarthritis

丝素蛋白小分子药物载体治疗骨关节炎

• 批准号: 8400807
• 负责人: Timothy Kariithi Mwangi
• 金额: $ 3.16万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-08-01 至 2015-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8400807
• 关键词: Address; Adopted; Affect; Alkylation; Amines; Amino Acids; Apoptosis; Appearance; Attention; Attenuated; Biocompatible Materials; Biological Availability; Biological Markers; Biomedical Engineering; Biomedical Research; Blood Circulation; Bombyx; Bombyx mori; Carbamates; Cell Line; Cells; Characteristics; Chemicals; Chondrocytes; Clinical; Coupled; Coupling; Curcumin; Data; Degenerative polyarthritis; Development; Disease; Disease Progression; Dose; Drug Carriers; Drug Delivery Systems; Fibroins; Filament; Fluorescence; Goals; Half-Life; High Pressure Liquid Chromatography; Histopathology; In Vitro; Inflammation; Inflammatory; Injectable; Injection of therapeutic agent; Intra-Articular Injections; Joint Instability; Joints; Knee joint; Label; Laboratories; Learning; Limb structure; Lymphocyte; Measures; Mentors; Methods; Modeling; Modification; Molecular Weight; Operative Surgical Procedures; Oral; Outcome; Pain; Particle Size; Pharmaceutical Preparations; Phenols; Phospholipids; Plasma; Pre-Clinical Model; Proteins; Rattus; Recording of previous events; Research; Safety; Science; Serine; Serum; Silk; Solubility; Suspension substance; Suspensions; Symptoms; Synovial Fluid; Therapeutic; Tumor Necrosis Factor Receptor; Tyrosine; Variant; Weight-Bearing state; Work; allodynia; anakinra; base; carboxylate; career; cell type; cost; cytokine; doctoral student; drug efficacy; experience; fluorescence imaging; human subject; in vivo; interest; joint injury; mechanical allodynia; nano; nanoparticle; nanoparticulate; novel; particle; pre-doctoral; response; small molecule; training project

DESCRIPTION (provided by applicant): Osteoarthritis (OA) is a degenerative joint disease associated with impaired activity and pain for the affected joints. Intra-articular injection of smll molecule drugs has great potential to treat both disease progression and symptoms of OA, but fails due to the very rapid clearance of small drugs from the joint space. Previous work in the Sponsor laboratory has advanced use of injectable drug depots for intra-articular delivery of protein drugs that can provided for sustained and local release of drug. The goal of the current project is to develop injectable, intra-articular carriers for small-molecule drugs that have potential to treat OA. I have adopted methods to synthesize nano- and micro-particles from silk fibroin (SF) towards the goal of coupling to small molecule drugs. SF is attractive as a drug depot for its extensive history of safe clinical use, and it is inexpensive to prepare in large quantities. Curcumin is a small di-phenol that inhibits inflammation in many cell types and will be used as a model drug here. A very low solubility and oral bioavailability for curcumin is very limiting, however, as it is difficult to achieve nanomolar curcumin concentrations following oral dosing as high as 10 grams. In this study, we will work with a novel curcumin variant - monofunctional curcumin carbamate (MCC) - that was synthesized at Duke to allow for increased drug solubility and to provide a reactive amine for coupling to silk. The hypothesis of this project and training period is that intra-articular delivery of SF particles conjugated to MCC will enhance drug efficacy in pre-clinical models of OA, by prolonging drug half-lives in the joint space while retaining bioactivity. Work in Aim 1 will optimize SF modification to enable MCC coupling (SF-MCC) and synthesize two groups of drug carriers: (1) nanoparticles (< 1 micron); or (2) microparticles. For each group, I will: (a) study in vitro MCC release and bioactivity against IL-1b, (b) determine in vivo half-life of SF particles following delivery to the joint spac using in vivo fluorescence imaging; and (c) measure in vivo clearance of MCC following intra-articular injection of SF-MCC in plasma via HPLC. This work will demonstrate if nano- or microparticles provide a preferred drug depot for the delivery of a small molecule to the joint space. Work in Aim 2 will evaluate an ability for SF-MCC particles to attenuate disease development and pain-related sensitivity in a rat joint instability model of OA. Rats with OA will be treated with intra-articular injection of curcumin, MCC, SF-MCC, or SF particles alone, and I will measure and compare (a) hind limb weight-bearing; (b) pain-related sensitivity (mechanical allodynia); (c) histopathology grade; and (d) serum and synovial fluid biomarkers of OA. Results from these studies should reveal if local and sustained delivery of a modified curcumin and delivery vehicle has potential to modify disease or symptoms associated with OA. I will work with my Sponsor and interdisciplinary mentoring team to learn fundamentals and applications of materials science, drug delivery, pre-clinical model outcomes that will extend my interest in translational bioengineering and prepare me for an academic career in bioengineering. PUBLIC HEALTH RELEVANCE: This project will advance use of a safe and inexpensive biomaterial carrier for small molecule drug delivery to the joint space. Nano and microparticles of silk fibroin will be conjugated to a small molecule inflammatory antagonist, curcumin, and evaluated as a sustained release drug depot following injection into the knee joint for the treatment of osteoarthritis. A goal of this project is to provide novel learning in translational bioengineering in order to further the applicant's potential for a successful academic career in biomedical research.

描述（由申请人提供）：骨关节炎（OA）是一种与受影响关节的活动受损和疼痛相关的退化性关节疾病。 SMLL分子药物的关节内注射具有治疗疾病进展和OA症状的巨大潜力，但是由于关节空间从小药物中清除了很快，因此失败了。赞助商实验室的先前工作已先进使用可注射药物库，用于蛋白质药物的关节内递送，这些药物可以为持续和局部释放药物提供。当前项目的目的是开发具有有潜力治疗OA的小分子药物的可注射的，关节内载体。我采用了从丝纤维蛋白（SF）合成纳米和微粒的方法，以偶联到小分子药物。 SF作为药物仓库具有吸引力，它具有广泛的安全临床用途历史，并且大量准备很便宜。姜黄素是一种小型二苯酚，抑制许多细胞类型的炎症，将是 在这里用作模型药物。然而，姜黄素的溶解度和口服生物利用度非常低，但是由于口服剂量高达10克，很难实现纳摩尔姜黄素浓度。在这项研究中，我们将使用一种新型的姜黄素变体 - 单官能姜黄素氨基甲酸酯（MCC） - 在DUKE合成以允许增加药物溶解度，并提供可反应性胺与丝绸偶联。该项目和培训期的假设是，与MCC结合的SF颗粒的关节内递送 将通过延长关节中的药物半衰期来增强OA前临床模型的药物疗效 在保持生物活性的同时空间。 AIM 1的工作将优化SF修饰以实现MCC耦合（SF-MCC）并合成两组药物载体：（1）纳米颗粒（<1 micron）；或（2）微粒。对于每组，我将：（a）使用体内荧光成像在输送到关节SPAC后，在体外MCC释放和针对IL-1B的生物活性研究；（b）确定SF颗粒的体内半衰期； （c）测量通过HPLC在等离子体中关节内注射SF-MCC后MCC的体内清除。这项工作将证明纳米或微粒是否提供了首选的药物库，用于将小分子递送到关节空间。 AIM 2的工作将评估SF-MCC颗粒在OA的大鼠关节不稳定性模型中减弱疾病发育和与疼痛相关的敏感性的能力。与OA的大鼠将通过关节内注射姜黄素，MCC，SF-MCC或SF颗粒进行治疗，我将测量和比较（a）后肢重量； （b）与疼痛相关的敏感性（机械异常症）； （c）组织病理学等级； （d）OA的血清和滑液生物标志物。这些研究的结果应揭示改良的姜黄素和递送工具的局部和持续递送是否有可能改变与OA相关的疾病或症状。我将与赞助商和跨学科指导团队合作，学习材料科学，药物输送，临床前模型成果的基础知识和应用，这将扩大我对翻译生物工程的兴趣，并为我为生物工程学的学术生涯做好准备。 公共卫生相关性：该项目将推动使用安全且廉价的生物材料载体将小分子药物输送到联合空间。丝质纤维蛋白的纳米和微粒将与小分子炎症拮抗剂，姜黄素结合，并在注射到膝关节后作为持续释放的药物库评估，以治疗骨关节炎。该项目的一个目标是在翻译生物工程中提供新颖的学习，以进一步申请人在生物医学研究中成功的学术生涯的潜力。