Therapeutic Interventions for HIV-1 CNS Sequestration.

描述（由申请人提供）：HIV-1在初次感染后进入中枢神经系统（CNS），并导致患者观察到的神经功能缺损。由于大多数抗逆转录病毒药物无法在大脑中达到足够的水平，因此在中枢神经系统中发现了中等到高的病毒滴度。血脑屏障（BBB）具有紧密连接和多种多药耐药（MDR）转运蛋白，如p -糖蛋白（Pgp）和多药耐药相关蛋白（MRP-2），可显著减少脑内抗逆转录病毒药物向大脑的输送。因此，这项治疗干预建议的假设是，通过可逆地打开紧密连接和调节血脑屏障中p -糖蛋白（MDR）介导的外排，可以增强向大脑的抗逆转录病毒药物递送，从而控制hiv -1相关的神经侵入。为了验证这一假设，我们将追求以下三个特定目的：利用细胞培养技术和SCID小鼠研究闭塞带毒素（Zot）增强血脑屏障运输、脑摄取和两种“模型”抗逆转录病毒药物（AZT, zidovudine）活性的能力。初步研究表明，Zot能够可逆地打开血脑屏障中的紧密连接，并显著（p < 0.05）增加脑内亲疏水药物（如紫杉醇、阿霉素）的水平。将使用未感染的SCID小鼠和在脑尾状核和壳核中接种了HIV-1感染单核细胞的SCID小鼠。脑部抗逆转录病毒水平和病毒滴度将被测定。利用细胞培养技术和SCID小鼠模型研究MDR对两种“模型”抗逆转录病毒药物AZT和沙奎那韦（MRP-1和Pgp的底物）的血脑屏障转运、脑摄取和活性的影响。采用细胞培养技术和SCID小鼠SA3，研究MDR对AZT和沙奎那韦（MRp-2和Pgp的底物）体外血脑屏障转运和脑分布的影响。研究Zot和MDR抑制剂对SCID小鼠血脑屏障转运、AZT和沙奎那韦的脑摄取和活性的伴随作用。获得治疗性中枢神经系统的抗逆转录病毒药物水平对于最小化中枢神经系统中的HIV-1至关重要。因此，这项建议为解决艾滋病毒/艾滋病这一并发症提供了一个可行的办法。

DESCRIPTION (provided by applicant): HIV-1 enters the Central Nervous System (CNS) after the primary infection and contributes to the neurological deficits observed in patients. Moderate to high viral titers are found in the CNS due to the fact that the majority of anti-retroviral agents are unable to achieve sufficient levels in the brain. The Blood Brain Barrier (BBB) possesses tight junctions and various Multi-Drug Resistance (MDR) transporters such as P-glycoprotein (Pgp) and Multi-drug Resistance Associated Protein (MRP-2), which serve to significantly minimize brain anti-retroviral drug delivery to the brain. Thus, the hypothesis of this therapeutic intervention proposal is that anti-retroviral drug delivery to the brain can be enhanced in order to manageHIV-1 related neuroinvasion by reversibly opening tight junctions and by modulation of MDR (P-glycoprotein,MRP) mediated efflux at the BBB. In order to test this hypothesis, the following three specific aims (SA) will be pursued SA1 To examine the ability of Zonula Occludens Toxin (Zot) to enhance the BBB transport, brain uptake and activity of two "model" antiretroviral agents (AZT, zidovudine) using cell culture techniques and the SCID mice. Preliminary studies have shown that Zot is capable of reversibly opening tight junctions in the BBB, as well as significantly (p < 0 05) increasing brain levels of hydrophilic and hydrophobic agents (e g, paclitaxel, doxorubicin).Uninfected SCID mice and SCID mice inoculated with HIV-1 infected monocytes in the brain caudate and putamen will be used. Antiretroviral brain levels and viral titers will be determined. SA 2 To examine the effect of MDR modulation on the BBB transport, brain uptake and activity of AZT and saquinavir (substrates of MRP-1 and Pgp) two "model" anti-retroviral agents using cell culture techniques and the SCID mouse model. To examine the effect of MDR modulation on the in vitro BBB transport and brain distribution of on AZT and saquinavir (substrates of MRp-2 and Pgp)using cell culture techniques and the SCID mice SA3.To examine the concomitant effect of Zot and the MDR inhibitors on the BBB transport, brain uptake and activity of AZT and saquinavir using SCID mice. Obtaining therapeutic CNS levels of anti-retroviral agents is essential to minimizing HIV-1 in the CNS. As such, this proposal offers a viable approach to address this complication of HIV/AIDS.