SBIR Phase I: Bioengineered Recombinant Anticoagulant Heparin

这个小企业创新研究（SBIR）一期项目的更广泛的影响/商业潜力将是利用工程培养的哺乳动物细胞开发药物肝素的替代来源。肝素是一种广泛使用的抗凝剂（血液稀释剂），对外科手术和有凝血风险的患者至关重要。仅在美国，每年就有超过30万剂疫苗被注射。目前，肝素是由动物组织制备的，在中国主要是由猪肠制备的，在美国和其他相关国家都有监管机构。一个原因是，肝素的生产在中国很难监管，2008年肝素掺假危机就证明了这一点，该危机导致了过敏反应，全球超过250人死亡。此外，有人担心中国的生猪数量将无法满足日益增长的需求。FDA正在考虑重新引入从肉牛中生产的肝素作为替代来源，然而，牛肝素仍然受到涉及动物组织的并发症的影响，并且它具有不同的抗凝血特性，这使得剂量复杂化。目前的建议是开发基于细胞的生产，其中整个供应链可以在GMP控制下进行，并且可以根据需要进行扩展，而不依赖于动物种群的数量和健康。这个SBIR一期项目的智力优势是通过基因工程细胞来生产可用于制药的肝素产品。细胞生产可以完全在GMP控制下，细胞工程提供了生产具有改进性能的肝素的机会。一项对使用肝素的临床医生的调查压倒性地确定了肝素诱导的血小板减少症（HIT）是一个严重的有害副作用。HIT可导致血栓、中风、心脏病发作以及生命和肢体的丧失。治疗既复杂又昂贵。HIT是一种过敏反应，当肝素与血液中称为血小板因子4 （PF4）的特定蛋白质结合时引起的。PF4的结合取决于肝素结构中不参与肝素抗凝活性的元素。通过细胞工程，将产生结构改变的肝素分子，降低对PF4的亲和力，同时保持有效的抗凝血特性。此外，肝素已被证明对其他领域有效，如炎症和癌症，不涉及血液凝固。通过细胞工程，肝素结构也可以为这些额外的应用量身定制。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

The broader impact/commercial potential of this Small Business Innovation Research (SBIR) Phase I project will be to develop an alternative source of pharmaceutical heparin using engineered cultured mammalian cells. Heparin is a widely prescribed anticoagulant (blood thinner), critical for surgical operations and for patients that otherwise have risks of blood clotting. Over 300,000 doses are administered per year in the US alone. Currently, heparin is prepared from animal tissues, primarily pig intestines in China, which has regulatory agencies in the US and other countries concerned. For one reason, heparin manufacturing is difficult to regulate in China as evidenced by the heparin adulteration crisis in 2008 that led to allergic reactions and over 250 deaths worldwide. In addition, there is concern that the Chinese pig population will not be able to keep up with the increasing demand. The FDA is considering reintroducing heparin produced from beef cattle as an alternative source, however, bovine heparin is still subject to complications involving animal tissues and it has different anticoagulant properties, which complicates dosing. The current proposal develops cell-based production in which the entire supply chain can be under GMP control and scalable as needed with no dependence on the number and health of an animal population.The intellectual merit of this SBIR Phase I project is to genetically engineer cells to produce a heparin product that can be manufactured for pharmaceutical use. Cellular production can be entirely under GMP control, and cell engineering provides opportunities to produce heparin with improved properties. A survey of clinicians that use heparin overwhelmingly identified heparin induced thrombocytopenia (HIT) as a serious detrimental side effect. HIT can lead to blood clots, stroke, heart attack and loss of life and limb. Treatment is complicated and expensive. HIT is an allergic reaction caused when heparin binds to a specific protein in the blood called platelet factor 4 (PF4). PF4 binding depends on elements of heparin structure that are not involved in heparin's anticoagulant activities. Through cell engineering, heparin molecules with an altered structure will be produced that have reduced affinities for PF4, while maintaining potent anticoagulant properties. In addition, heparin has demonstrated efficacy in other areas such as inflammation and cancer that are not involved in blood clotting. Through cell engineering, heparin structures also could be tailored for these additional applications.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.