TARGETED DRUG RELEASE IN VIRUS-INFECTED MACROPHAGES

描述(改编自调查员摘要)：调查员请求 为第一阶段SBIR提供资金，以制定有针对性的交付方法 巨噬细胞的抗逆转录病毒药物。最终，包覆的微粒 含有载脂前体药物的巨噬细胞在体外会被摄取， 微粒包衣会产生不稳定的水解性，使脂类成分 将通过HIV或CMV病毒蛋白水解酶-可消化多肽连接。这个 脂质部分将以神经酰胺为基础，以促进亚细胞定位 与巨噬细胞内富含病毒的高尔基体区域的脂质连接的前药。 如果巨噬细胞感染了HIV或CMV，则相应的病毒蛋白水解酶 会将脂质部分从前药中分离出来并释放出活性形式 感染部位的药物。现拟进行的第一阶段研究 格兰特将使用模型染料-多肽连接和体外模拟。释放 与HIV或HIV孵育后微粒中的荧光染料 CMV蛋白酶将提供概念证明，并为第二阶段铺平道路 后续行动。 AIM 1将开发整合病毒的合成程序 蛋白水解酶特异的、可切割的结合到药物结合化学中以及 脂类染料络合物的高效液相色谱分析方法。最初， 神经酰胺-多肽(HIV)-染料和神经酰胺-多肽(CMV)-染料复合体 做好准备。目标2将确定模型染料从 当与HIV和CMV酶孵育时，脂类染料复合体。重组HIV 将购买蛋白酶，由Scott Wong博士提供重组人巨细胞病毒蛋白酶 (ORPC)分包合同。这两个目标都是为目标3做准备，其中 脂类染料复合体将被用来包裹不同大小的微粒。 最后，在目标4中，调查人员将确定 为微粒提供不稳定的水解性涂层，以防止早产 在吞噬巨噬细胞之前释放染料。 建议的商业应用：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