Control of Drug Delivery to the CNS: Modulating P-glycoprotein Activity

描述（由申请人提供）： 项目研究和开发的概要说明（项目的主要开发商Banks，William A Title）（不超过72个字符空间）控制药物向CNS的递送：调节P-糖蛋白活性关键词（仅限MeSH术语;最少三种）药物、血脑屏障、脑、CNS、p-糖蛋白、先天免疫系统、细胞因子、肿瘤坏死因子-α，一氧化氮研究进展简述（不要使用续页）在开发有效的脑治疗方法中的一个主要问题是血脑屏障（BBB），它将许多药物排除在中枢神经系统（CNS）之外。BBB排除药物的一个主要机制是通过P-糖蛋白（P-gp）。P-gp是位于BBB的管腔侧的脑-血（流出）泵，其防止其配体进入CNS。P-gp对大量临床使用的药物具有亲和力。增加P-gp活性进一步限制其配体进入CNS，从而使许多CNS药物无效，而降低P-gp活性允许更多药物进入CNS，从而增加作用。我们的长期目标是了解如何控制P-gp活性，从而控制其配体向CNS的递送。最近在体外脑内皮细胞的工作已经证明了级联的事件，调节P-gp活性。脑内皮细胞上的P-gp被脂多糖（LPS）和肿瘤坏死因子α（TNF）抑制。这些依次刺激内皮素-1（ET-1），内皮素-1通过其B受体（ETR B）起作用以增加一氧化氮合酶（NOS）活性，所述一氧化氮合酶（NOS）活性在数分钟内起作用以降低P-gp活性达min-h：LPS -> TNF -> ET-1（ETR B）->NOS ->降低的P-gp活性。该途径还通过ET-1受体ETRA和ETRB刺激NF κ B以增加P-gp的表达，在数小时内产生P-gp的长期刺激：TNF -> ET-1（ETRA/ETRB）-> NOS -> NF κ B-> mRNA（P-gp）->增加的P-gp活性。 我们的目标是确定这些机制是否在体内运作。我们的工作假设是：P-gp活性在体内受TNF、ET-1和NOS顺序激活的调节。我们的具体目标是：SA 1：在小鼠体内确定TNF对P-gp的短期和长期影响。SA 2：在小鼠体内确定ET-1对LPS诱导的P-gp变化的作用。SA 3：在小鼠体内测定NOS对P-gp的短期和长期影响。对退伍军人医疗保健的潜在影响：如果该级联的全部或部分在体内具有活性，则P-gp活性可能在许多疾病中发生改变，从而导致CNS药物活性改变。这些疾病不仅包括细菌性败血症，还包括诸如类风湿性关节炎、糖尿病、中风、CNS癌症、甚至肥胖症和阿尔茨海默病等疾病，其中存在促炎性和神经炎性病症。这项工作将对医疗保健产生3个主要影响：1）它将有助于确定药物和疾病，其中修饰的P-gp活性导致CNS药物谱改变; 2）它将确定可以逆转这些疾病状态下CNS药物谱改变的策略和药物; 3）这将为开发靶向治疗中枢神经系统疾病的药物和开发副作用小的药物提供一种新的策略，即调控P-gp活性。VA表10-1313-2 VA表10-1313包装第2页1990年6月（R） 公共卫生相关性： 对退伍军人医疗保健的潜在影响：如果该级联的全部或部分在体内是活性的，那么P-gp活性可能在许多疾病中被改变，这将导致CNS药物活性改变。这些疾病不仅包括细菌性败血症，还包括诸如类风湿性关节炎、糖尿病、中风、CNS癌症、甚至肥胖症和阿尔茨海默病等疾病，其中存在促炎性和神经炎性病症。这项工作将对医疗保健产生3个主要影响：1）它将有助于确定药物和疾病，其中修饰的P-gp活性导致CNS药物谱改变; 2）它将确定可以逆转这些疾病状态下CNS药物谱改变的策略和药物; 3）这将为开发靶向治疗中枢神经系统疾病的药物和开发副作用小的药物提供一种新的策略，即调控P-gp活性。

DESCRIPTION (provided by applicant): SUMMARY DESCRIPTION OF PROGRAM SUMMARY DESCRIPTION OF PROJECT RESEARCH AND DEVELOPMENT PROGRAM PRINCIPAL INVESTIGATOR(S) Banks, William A TITLE OF PROGRAM PROJECT (Not to exceed 72 character spaces) Control of Drug Delivery to the CNS: Modulating P-glycoprotein Activity KEYWORDS (MeSH terms only; minimum three) Drug, blood-brain barrier, Brain, CNS, p-glycoprotein, innate immune system, cytokines, tumor necrosis factor-alpha, nitric oxide BRIEF STATEMENT OF RESEARCH OBJECTIVES (Do not use continuation sheets) A major problem in developing effective therapeutics for the brain is the blood-brain barrier (BBB), which excludes many drugs from the central nervous system (CNS). One major mechanism by which the BBB excludes drugs is through P- glycorprotein (P-gp). P-gp is a brain-to-blood (efflux) pump located in the luminal side of the BBB that prevents its ligands from entering the CNS. P-gp has an affinity for an incredibly large number of clinically used drugs. Increasing P- gp activity further limits the access of its ligands to the CNS, thus making many CNS drugs ineffective, whereas decreasing P-gp activity allows more drug into the CNS, thus increasing effects. Our long-term goal is to learn how to control P-gp activity and so control the delivery of its ligands to the CNS. Recent in vitro work with brain endothelial cells has demonstrated a cascade of events that regulate P-gp activity. P-gp at the brain endothelial cell is inhibited by lipopolysaccharide (LPS) and tumor necrosis factor alpha (TNF). These sequentially stimulate endothelin-1 (ET-1), which acts through its B receptor (ETRB) to increase nitric oxide synthase (NOS) activity that acts within minutes to decrease P- gp activity for min-h: LPS -> TNF -> ET-1 (ETRB) ->NOS -> decreased P-gp activity. This pathway also stimulates NFkB through the ET-1 receptors ETRA and ETRB to increase expression of P-gp, producing within hours a long-term stimulation of P-gp: TNF -> ET-1 (ETRA/ETRB) -> NOS -> NFkB -> mRNA (P-gp) -> increased P-gp activity. Our goal is to determine whether these mechanisms are operational in vivo. Our working hypothesis is: P-gp activity is regulated in vivo by the sequential activation of TNF, ET-1, and NOS. Our specific aims are: SA 1: To determine in vivo in mice the short- and long-term effects of TNF on P-gp. SA2 : To determine in vivo in mice the role of ET-1 on LPS-induced change in P-gp. SA 3: To determine in vivo in mice the short- and long-term effects of NOS on P-gp. Potential Impact on Veteran's Health Care: If all or part of this cascade is active in vivo, then P-gp activity is likely modified in a host of diseases that would result in altered CNS drug activities. These diseases would include not only bacterial sepsis, but diseases such as rheumatoid arthritis, diabetes, stroke, CNS cancers, and even obesity and Alzheimer's disease in which proinflammatory and neuroinflammatory conditions exist. The proposed work will have 3 major impacts on health care: 1) it will help to identify drugs and diseases in which a modified P-gp activity results in altered CNS drug profiles; 2) it will identify stategies and agents which can reverse those altered CNS drug profiles in those disease states; 3) it will lead to a new strategy, that of manipulation of P-gp activity, in the development of drugs both for targeting CNS diseases and for developing drugs with fewer CNS side effects. VA FORM 10-1313-2 Page 2 of VA Form 10-1313 package JUN 1990(R) PUBLIC HEALTH RELEVANCE: Potential Impact on Veterans' Health Care: If all or part of this cascade is active in vivo, then P-gp activity is likely modified in a host of diseases that would result in altered CNS drug activities. These diseases would include not only bacterial sepsis, but diseases such as rheumatoid arthritis, diabetes, stroke, CNS cancers, and even obesity and Alzheimer's disease in which proinflammatory and neuroinflammatory conditions exist. The proposed work will have 3 major impacts on health care: 1) it will help to identify drugs and diseases in which a modified P-gp activity results in altered CNS drug profiles; 2) it will identify stategies and agents which can reverse those altered CNS drug profiles in those disease states; 3) it will lead to a new strategy, that of manipulation of P-gp activity, in the development of drugs both for targeting CNS diseases and for developing drugs with fewer CNS side effects.