Therapeutic Interventions for HIV-1 CNS Sequestration.

描述（由申请方提供）：HIV-1在原发性感染后进入中枢神经系统（CNS），并导致在患者中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CNS中发现中度至高度病毒滴度。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），其用于显著地最小化脑抗逆转录病毒药物递送至脑。因此，该治疗干预建议的假设是，可以增强抗逆转录病毒药物向脑的递送，以便通过可逆地打开紧密连接和通过调节MDR（P-糖蛋白，MRP）介导的BBB流出来管理HIV-1相关的神经侵袭。为了验证这一假设，将追求以下三个具体目标（SA）SA 1使用细胞培养技术和SCID小鼠来检查闭锁小带毒素（Zot）增强BBB转运、脑摄取和两种“模型”抗逆转录病毒药物（AZT、齐多夫定）活性的能力。初步研究表明，Zot能可逆性地开放血脑屏障的紧密连接，并显著（p <0.05）增加脑内亲水性和疏水性物质（如紫杉醇、阿霉素）的水平。将测定抗逆转录病毒脑水平和病毒滴度。SA 2利用细胞培养技术和SCID小鼠模型，研究MDR调节对AZT和沙奎那韦（MRP-1和Pgp的底物）两种抗逆转录病毒药物的BBB转运、脑摄取和活性的影响。采用细胞培养技术和SCID小鼠SA 3模型，观察MDR对AZT和Saquinavir（MRp-2和Pgp的底物）体外血脑屏障转运和脑分布的影响。采用SCID小鼠，观察Zot和MDR抑制剂对AZT和Saquinavir血脑屏障转运、脑摄取和活性的协同作用。获得治疗性CNS水平的抗逆转录病毒药物对于最大限度地减少CNS中的HIV-1至关重要。因此，这项建议为解决艾滋病毒/艾滋病的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.