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

 产品说明：在当前系统中生产的可注射生物药物过于昂贵，全球大多数人口负担不起。最近的研究表明，植物细胞中的蛋白质药物的生物包囊在胃中受到保护。共生微生物在其释放到肠腔中中起关键作用。将药物生物包封的植物细胞快速递送到循环中，组织学证据和抗生素研究表明小肠微生物在这一过程中发挥了关键作用。我们还确定了独特的标签，可以在穿过肠道上皮后将治疗性蛋白质递送到循环、非免疫或免疫调节细胞。 该提案解决了口服递送人血蛋白中的一些剩余挑战，使得这种新概念可以推进到临床。蛋白质药物应在可食用植物细胞中而不是烟草中表达，并应提高其表达水平（特别是对于大蛋白），以促进剂量递增研究。为了实现这一目标，将使用先进的现代工具。在叶绿体中表达的天然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替换稀有密码子。GLP-1样短肽（毒蜥外泌肽）或血管紧张素1-7的表达水平将通过串联重复序列和具有普遍存在的蛋白酶（如弗林蛋白酶）的切割位点而增强。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，并将使用合适的动物模型评价其治疗高血压/糖尿病或诱导血友病口服耐受的功能。肠道微生物的作用将通过人类小肠细菌物种的功能表征和鉴定来研究，这些细菌物种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。提出了在cGMP工厂（Fraunhofer USA，特拉华州）生产成功的候选物、在环境温度下长期储存后的稳定性评价、有效性和剂量。 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递送和延长蛋白质药物的保质期来显著降低医疗保健成本之外，还应提高患者的依从性。这一概念的临床进展将彻底改变蛋白质药物的生产和交付大多数代谢和遗传疾病。与充分研究的粪便/结肠系统相反，人类小肠微生物群的功能表征先前尚未被探索，并且该研究将促进对人类肠道的进一步机械理解以及肠道微生物在植物细胞加工和利用人体中最大吸收表面递送蛋白质药物中的作用。

 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.