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

 描述：在目前的系统中生产的可注射生物药物价格高得令人望而却步，而且对全球大多数人口来说负担不起。最近的研究表明，蛋白质药物在植物细胞中的生物胶囊在胃中具有保护作用。共生微生物在它们释放到肠腔中起着关键作用。生物包裹植物细胞的药物快速进入循环，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着关键作用。我们还识别了独特的标签，可以在穿过肠道上皮后将治疗性蛋白输送到循环、非免疫或免疫调节细胞。这项建议解决了口服人类血液蛋白的一些剩余挑战，以便将这一新概念推向临床。蛋白质药物应该在可食用的植物细胞中表达，而不是在烟草中表达，并且它们的表达水平应该提高(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，将使用尖端的现代工具。在叶绿体中表达的天然人类基因的核糖体图谱将确定核糖体停顿位置。密码子优化将用最优密码子替换稀有密码子。GLP-1类短肽(Exendin)或血管紧张素1-7的表达水平将通过串联重复序列和具有普遍存在的蛋白酶(如Furin)的裂解位点而增强。将对不同的递送标签进行测试，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估其用于治疗高血压/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对能够释放植物细胞中生物包裹的蛋白质药物和宿主吸收的人类小肠细菌的功能鉴定和鉴定来研究。提出了在cGMP设施(美国特拉华州弗劳恩霍夫)生产成功的候选者、常温下长期储存的稳定性评价、有效性和剂量。这些新方法除了通过取消昂贵的发酵罐、纯化、冷藏/运输、当前使用的方法的无菌交付以及延长蛋白质药物的保质期，还应显著降低医疗保健成本，还应提高患者的依从性。这一概念的临床进展将彻底改变大多数代谢和遗传疾病的蛋白质药物的生产和递送。与研究良好的粪便/结肠系统相比，以前还没有人探索过人类小肠微生物区系的功能特征，这项研究将有助于从机制上进一步了解人类肠道以及肠道微生物在处理植物细胞和利用人体最大吸收表面输送蛋白质药物方面的作用。

 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.