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: 9340258
• 负责人: HENRY DANIELL
• 金额: $ 68.83万
• 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/9340258
• 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; Drug Utilization; Edible Plants; Encapsulated; Engineering; Enzymes; Epithelial; Evaluation; Excision; Fermentation; Freeze Drying; Genes; Glycosides; Health Care Costs; Hemophilia A; Hereditary Disease; Histologic; Human; Human body; Hypertension; Immune; Immune Tolerance; Immune system; Injectable; Insulin; Intestines; Large Intestine; Lettuce - dietary; Life; Metabolic; Metabolic Diseases; Methods; Microbe; Modernization; Myelogenous; Oral; Patient Care; Peptide Hydrolases; Peptides; Pharmaceutical Preparations; Plants; Play; Polysaccharides; Population; Process; Production; Property; Proteins; Publications; Reporting; Reproducibility; Ribosomes; Role; Sampling; Site; Sterility; Stomach; Surface; System; T-Lymphocyte; Tandem Repeat Sequences; Temperature; Testing; Therapeutic; Tissues; Tobacco; Toxic effect; Transportation; absorption; base; cell type; clinical translation; 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：当前系统生产的注射生物药品价格昂贵，全球大多数人无法承受。最近的研究表明，植物细胞中蛋白质药物的生物封装在胃中受到保护。共生微生物在释放到肠腔中发挥着关键作用。将生物封装的植物细胞药物快速输送到循环系统中，组织学证据和抗生素研究表明小肠微生物在此过程中发挥着关键作用。我们还发现了独特的标签，可以在穿过肠道上皮后将治疗蛋白递送至循环细胞、非免疫细胞或免疫调节细胞。 该提案解决了人类血液蛋白口服输送中剩余的一些挑战，以便将这一新概念推进到临床。蛋白质药物应在可食用植物细胞而不是烟草中表达，并且应提高其表达水平（特别是大蛋白质）以促进剂量递增研究。为了实现这一目标，将利用尖端的现代工具。对叶绿体中表达的天然人类基因进行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替代稀有密码子。 GLP-1（如短肽（exendin）或血管紧张素 1-7）的表达水平将通过串联重复序列和普遍存在的蛋白酶（如弗林蛋白酶）的切割位点得到增强。将测试不同的递送标签以将蛋白质靶向非免疫或免疫调节细胞，并使用合适的动物模型评估其功能以治疗高血压/糖尿病或诱导血友病的口服耐受。将通过功能表征和鉴定能够释放生物封装在植物细胞中的蛋白质药物和宿主吸收的人类小肠细菌种类来研究肠道微生物的作用。提出了在 cGMP 设施（美国特拉华州弗劳恩霍夫）生产成功候选产品的候选方案，并提出了在环境温度下长期储存的稳定性、功效和剂量的评估。 这些新方法除了通过消除昂贵的发酵罐、纯化、冷藏/运输、目前使用的方法的无菌输送以及延长蛋白质药物的保质期来显着降低医疗成本之外，还应该提高患者的依从性。这一概念的临床进步将彻底改变大多数代谢和遗传疾病的蛋白质药物生产和输送。与经过充分研究的粪便/结肠系统相比，人类小肠微生物群的功能特征此前尚未被探索过，这项研究将有助于进一步了解人类肠道的机制以及肠道微生物在植物细胞加工和利用人体最大吸收表面输送蛋白质药物中的作用。