Therapeutic Interventions for HIV-1 CNS Sequestration.

描述(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障碍。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，在中枢神经系统中发现了中到高的病毒滴度。血脑屏障(BBB)具有紧密的连接和多种多药耐药(MDR)转运体，如P-糖蛋白(Pgp)和多药耐药相关蛋白(MRP-2)，可显著减少脑部抗逆转录病毒药物的脑内转运。因此，这一治疗干预方案的假设是，通过可逆地开放紧密连接和调节MDR(P-糖蛋白，MRP)介导的血脑屏障外流，可以增强抗逆转录病毒药物对大脑的输送，从而管理HIV-1相关的神经侵袭。为了验证这一假设，以下三个特定目标(SA)将被追求SA1，以检查闭锁小带毒素(Zot)是否能够利用细胞培养技术和SCID小鼠来增强两种“模型”抗逆转录病毒药物(AZT，Zidovudine)的血脑屏障转运、脑摄取和活性。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&lt；005)增加大脑中亲水性和疏水性物质(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HIV-1感染单核细胞的SCID小鼠。将确定抗逆转录病毒脑水平和病毒滴度。采用细胞培养技术和SCID小鼠模型，观察MDR调节对AZT和沙奎那韦(MRP-1和PGP的底物)两种“模型”抗逆转录病毒药物的血脑屏障转运、脑摄取和活性的影响。采用细胞培养技术和SCID小鼠SA3模型，观察MDR调控对AZT和沙奎那韦(MRP-2和Pgp的底物)血脑屏障转运和脑内分布的影响。获得中枢神经系统治疗水平的抗逆转录病毒药物对于最大限度地减少中枢神经系统中的艾滋病毒-1至关重要。因此，这项提议为解决艾滋病毒/艾滋病的这一复杂问题提供了一种可行的办法。

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.