TARGETED DRUG RELEASE IN VIRUS-INFECTED MACROPHAGES

病毒感染的巨噬细胞中的靶向药物释放

• 批准号: 2784262
• 负责人: Takuji Tsukamoto
• 金额: $ 10万
• 依托单位: CHEMICA TECHNOLOGIES, INC.
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-09-30 至 2001-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2784262
• 关键词: antiviral agents; ceramides; cytomegalovirus; dosage forms; drug delivery systems; endopeptidases; fluorescent dye /probe; human immunodeficiency virus; macrophage; microcapsule; prodrugs; proteolysis; technology /technique development; virus infection mechanism; virus protein

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

描述（改编自研究者的摘要）：研究者要求 第一阶段 SBIR 的资金，用于开发有针对性的交付方法 针对巨噬细胞的抗逆转录病毒药物。最终，包被的微粒 含有脂质的前药在体外会被巨噬细胞摄取， 微粒涂层会水解不稳定，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。这 脂质部分将以神经酰胺为基础，以促进亚细胞定位 脂质连接的前药与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，相应的病毒蛋白酶 将从前药中裂解脂质部分并释放活性形式 感染部位的药物。目前提出的第一阶段研究 格兰特将使用模型染料-肽连接和体外模拟。释放 与 HIV 一起孵育后，微粒中的荧光染料或 CMV 蛋白酶将提供概念验证并为第二阶段铺平道路 后续行动。 目标 1 将开发用于掺入病毒的合成程序 蛋白酶特异性、可裂解的键进入药物附着化学物质以及 用于分析脂质-染料复合物的 HPLC 程序。最初， 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 与 HIV 和 CMV 蛋白酶一起孵育时的脂质染料复合物。重组艾滋病病毒 购买蛋白酶，rhCMV 蛋白酶由 Dr. Scott Wong 提供 （ORPC）根据分包合同。这两个目标都是为目标 3 做准备的，其中 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 微粒的水解不稳定涂层可防止过早发生 巨噬细胞摄入前染料释放。 拟议的商业应用：不可用