TARGETED DRUG RELEASE IN VIRUS-INFECTED MACROPHAGES

描述（改编自研究者摘要）：研究者要求 为第一阶段SBIR提供资金，以制定有针对性的交付方法， 抗逆转录病毒药物给巨噬细胞。最终， 含有脂质的前药将在体外被巨噬细胞摄取， 微粒涂层将是水解不稳定的，并且脂质部分 将通过HIV或CMV病毒蛋白酶可消化的肽键连接。的 脂质部分将是基于神经酰胺的，以促进 将脂质连接的前体药物转移到巨噬细胞内富含病毒的高尔基体区域。 如果巨噬细胞感染了HIV或CMV， 将从前药上裂解脂质部分并释放活性形式的 药物在感染部位。目前提出的I期研究 格兰特将使用模型染料肽连接和体外模拟。释放 与HIV孵育后来自微粒的荧光染料，或 CMV蛋白酶将提供概念验证并为II期铺平道路 跟进。 目标1将开发用于掺入病毒的合成程序， 蛋白酶特异性的、可裂解的键结合成药物附着化学物质， 用于分析脂质-染料-复合物的HPLC程序。起初， 神经酰胺-多肽（HIV）-染料和神经酰胺-多肽（CMV）-染料复合物将 做好准备目标2将确定模型染料从聚合物中的释放速率。 当与HIV和CMV蛋白酶孵育时，脂质-染料复合物。重组HIV 蛋白酶将购买，rhCMV蛋白酶将由Scott Wong博士提供 （ORPC）根据《公约》。这两个目标都是目标3的预备性目标， 脂质-染料-复合物将用于涂覆不同尺寸的微粒。 最后，在目标4中，研究人员将确定 用于微粒的水解不稳定涂层， 在巨噬细胞摄取之前释放染料。 拟议商业应用：不可用

DESCRIPTION (adapted from investigator's abstract): The investigators request funds for a Phase I SBIR to develop targeted delivery methods for anti-retroviral drugs to macrophages. Eventually, coated microparticles which contain lipid-bearing prodrugs will be ingested by macrophages in vitro, the microparticle coating will be hydrolytically unstable, and the lipid moieties will be linked via a HIV or CMV viral protease-digestible peptide linkage. The lipid moiety will be ceramide-based to facilitate subcellular localization of the lipid-linked prodrug to the virus-rich Golgi region within the macrophage. If the macrophages are infected with HIV or CMV, the respective viral proteases will cleave the lipid moiety from the prodrug and release the active form of the drug at the site of infection. The Phase I studies proposed in the present grant will use a model dye-peptide linkage and in vitro simulation. Release of the fluorescent dye from the microparticles following incubation with HIV or CMV proteases will provide proof of concept and pave the way for Phase II followup. Aim 1 will develop synthetic procedures for incorporation of viral protease-specific, cleavable bonds into drug attachment chemistries as well as HPLC procedures for analysis of the lipid-dye-complexes. Initially, ceramide-polypeptide(HIV)-dye and ceramide-polypeptide(CMV)-dye complexes will be prepared. Aim 2 will determine the rate of release of the model dye from the lipid-dye complex when incubated with HIV and CMV proteases. Recombinant HIV protease will be purchased, rhCMV protease will be provided by Dr. Scott Wong (ORPC) under subcontract. Both of these aims are preparatory to Aim 3 in which the lipid-dye-complexes will be used to coat microparticles of varying sizes. Finally, in Aim 4, the investigators will determine the best properties of the hydrolytically unstable coating for the microparticles to preclude premature dye release prior to macrophage ingestion. PROPOSED COMMERCIAL APPLICATION: NOT AVAILABLE