Rational design of potent recrystallization inhibitors

抗冻糖蛋白(AFGP)是一类天然存在的生物防冻剂。AFGP 1是最大的组分(33.7 KDa)，AFGP 8(2.6 KDa，图1)是最低的分子量组分。这些化合物主要存在于硬骨鱼中，通过防止体内种子冰晶的不受控制的生长来保护这些生物免受低温伤害和死亡。在过去的几年里，我们的实验室一直在积极参与合理设计和合成碳连接的(C连接的)AFGP类似物，具有增强的化学和生物稳定性以及增强的抗冻活性。这类化合物非常适合用于冷冻保存应用，因为它们不表现出任何细胞毒性，并且在体外抑制caspase酶。本提案中描述的研究将使用成熟的合成方法来合成一系列C连接的AFGP类似物，当测试活性时，将确定抑制组织中冰重结晶所必需的关键结构属性。有了这些信息，我们将能够设计出适用于医疗、商业和工业应用的最终的C-连接AFGP类似物。在这种跨学科环境中接受培训的研究生和本科生将获得科学实验、有机合成和细胞生物学方面的基本技能，为他们在生物制药和化学制造行业以及学术研究中的成功职业做准备。

Antifreeze glycoproteins (AFGPs) are a sub-class of naturally occurring biological antifreezes. AFGP 1 is the largest fraction (33.7 KDa) and AFGP 8 (2.6 KDa, figure 1) is the lowest molecular weight fraction. These compounds are found predominately in Teleost fish and protect these organisms against cryoinjury and death by preventing the uncontrolled growth of seeded ice crystals in vivo.      During the past several years our laboratory has been actively involved in the rational design and synthesis of carbon-linked (C-linked) AFGP analogues possessing enhanced chemical and biological stability as well as increased antifreeze activity.  This class of compounds is ideally suited for cryopreservation applications as they do not exhibit any cytotoxicity and inhibit caspase enzymes in vitro.  The studies described in this proposal will employ well-developed synthetic methodologies to synthesize a series of C-linked AFGP analogues which when tested for activity, will identify the key structural attributes necessary to inhibit the recrystallization of ice in tissues.  With this information we will be able to design the "ultimate" C-linked AFGP analogue suitable for medical, commercial and industrial applications.     Graduate and undergraduate students training in this interdisciplinary environment will acquire essential skills in scientific experimentation, organic synthesis and cell biology preparing them for successful careers in the biopharmaceutical and chemical manufacturing industries as well as academic research.