Affordable oral delivery of human blood protein drugs bioencapsulated in plant cells

以经济实惠的方式口服生物封装在植物细胞中的人血蛋白药物

• 批准号: 9107034
• 负责人: HENRY DANIELL
• 金额: $ 63.39万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-07-15 至 2020-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9107034
• 关键词: Acids; Address; Angiotensins; Animal Model; Antibiotic Therapy; Antibiotics; Antigen-Presenting Cells; Autoimmune Diseases; B-Lymphocytes; Binding; Binding Proteins; Biological Products; Blood Circulation; Blood Glucose; Blood Proteins; Carbohydrates; Cardiovascular system; Cell Wall; Cells; Chloroplasts; Cholera Toxin; Cleaved cell; Clinic; Clinical; Codon Nucleotides; Country; Cryopreservation; Cyclic GMP; Data; Delaware; Dendritic Cells; Diabetes Mellitus; Dose; Drug Delivery Systems; Edible Plants; Encapsulated; Engineering; Enzymes; Epithelial; Evaluation; Excision; Fermentation; Genes; Health Care Costs; Hemophilia A; Hereditary Disease; Human; Human body; Hypertension; Immune; Immune Tolerance; Immune system; Injectable; Insulin; Intestines; Large Intestine; Lettuce - dietary; Life; Metabolic; Metabolic Diseases; Methods; Microbe; Myeloid Cells; Oral; Patient Care; Peptide Hydrolases; Peptides; Pharmaceutical Preparations; Plants; Play; Polysaccharides; Population; Process; Production; Property; Proteins; Publications; Reporting; Ribosomes; Role; Sampling; Site; Sterility; Stomach; Surface; System; T-Lymphocyte; Tandem Repeat Sequences; Temperature; Testing; Therapeutic; Tissues; Tobacco; Toxic effect; Translations; Transportation; Villus; absorption; base; cell type; commensal microbes; compliance behavior; cost; dosage; drug production; gastrointestinal epithelium; gastrointestinal system; glucagon-like peptide 1; gut microbiota; histological studies; hypertension treatment; ileum; improved; intestinal epithelium; microbiota; novel; novel strategies; oral tolerance; public health relevance; receptor; receptor binding; ribosome profiling; scale up; targeted delivery; therapeutic protein; tool; vector

 DESCRIPTION: Injectable biopharmaceuticals produced in current systems are prohibitively expensive and are not affordable for a large majority of the global population. Recent studies have shown that bioencapsulation of protein drugs in plant cells are protected in the stomach. Commensal microbes play a critical role in their release into the gut lumen. Rapid delivery of drugs bioencapsulated plant cells into circulation, histological evidence and studies with antibiotics show the critical role played by small intestinal microbes in this process. We have also identified unique tags that could deliver therapeutic proteins to the circulation, non-immune or immune modulatory cells after crossing the gut epithelium. This proposal addresses some of the remaining challenges in oral delivery of human blood proteins so that this novel concept could be advanced to the clinic. Protein drugs should be expressed in edible plant cells instead of tobacco and their expression level should be enhanced (especially for large proteins) to facilitate dose escalation studies. In order to achieve this, cutting edge modern tools will be utilized. Ribosome profiling of native human genes expressed in chloroplasts will identify ribosome pause sites. Codon optimization will substitute rare codons with optimal codons. Level of expression of GLP-1 like short peptides (exendin) or angiotensin 1-7 will be enhanced with tandem repeats and cleavage sites with ubiquitous proteases like furin. Different delivery tags will be tested to target proteins to non-immune or immune modulatory cells and their functionality will be evaluated for treatment of hypertension/diabetes or induction of oral tolerance in hemophilia using suitable animal models. The role of gut microbes will be studied by functional characterization and identification of human small intestinal bacterial species capable of releasing protein drugs bio encapsulated in plant cells and host absorption. Production successful candidates in cGMP facility (Fraunhofer USA, Delaware), evaluation of stability upon prolonged storage at ambient temperature, efficacy and dosage are proposed. These novel approaches should improve patient compliance in addition to significantly lowering the cost of healthcare by elimination of prohibitively expensive fermenters, purification, cold storage/ transportation, sterile delivery of currently used methods and extending shelf life of protein drugs. Clinical advancement of this concept would revolutionize protein drug production and delivery for most metabolic and genetic disorders. In contrast to the well-studied fecal/colonic system, functional characterization of the human small intestinal microbiota has not been previously explored and this study would facilitate further mechanistic understanding of the human gut and the role of intestinal microbes in processing of plant cells and delivery of protein drugs utilizing the largest absorption surface in the human body.

 描述：在当前系统中生产的可注射的生物药物的昂贵，对于大多数全球人口来说都是不起作用的。最近的研究表明，植物细胞中蛋白质药物的生物塑造受到胃中的保护。共生微生物在将其释放到肠道内发挥着关键作用。药物的快速递送生物封装的植物细胞进入循环，组织学证据和抗生素研究表明，小肠道微生物在此过程中所起的关键作用。我们还确定了可以将治疗蛋白传递给循环，非免疫或免疫调节性细胞后的独特标签。该建议涉及人类血液蛋白口服递送的剩余挑战，以便可以将这种新颖的概念提升到诊所。蛋白质药物应在可食用的植物细胞而不是烟草中表达，应增强其表达水平（尤其是大蛋白），以促进剂量升级研究。为了实现这一目标，将利用最先进的现代工具。在叶绿体中表达的本地人类基因的核糖体分析将识别核糖体暂停位点。密码子优化将用最佳密码子代替稀有密码子。 GLP-1的表达水平，例如短肽（Exendin）或血管紧张素1-7，串联重复序列和裂解位点将增强，并具有无处不在的蛋白（如脂蛋白）。将测试不同的递送标签，以靶向蛋白质，以靶向非免疫或免疫调节细胞，其功能将被评估用于治疗高血压/糖尿病或使用合​​适的动物模型在血友病中诱导口服耐受性。肠道微生物的作用将通过功能表征和鉴定能够释放植物细胞中封装的蛋白质药物的人类小肠细菌物种并在CGMP设施（特拉华州Fraunhofer）中成功生产候选者，在储存量的稳定性中，以稳定的稳定性和宿主生产成功，并在环境温度下，有效，有效性，有效性。这些新颖的方法还应通过消除违禁昂贵的发酵罐，纯化，冷藏/运输，当前使用的方法的无菌递送以及延长蛋白质药物的保质期除外，还应提高患者依从性。该概念的临床发展将彻底改变大多数代谢和遗传疾病的蛋白质药物的产生和递送。与研究良好的粪便/结肠系统相反，以前尚未探索人类小肠道菌群的功能表征，这项研究将有助于进一步了解人类肠道的机械理解以及肠道植物细胞加工的作用以及利用人体中最大吸收表面的蛋白质药物的递送。