WO2020199144A1 - 阿尔兹海默症治疗靶点 - Google Patents

阿尔兹海默症治疗靶点 Download PDF

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WO2020199144A1
WO2020199144A1 PCT/CN2019/081167 CN2019081167W WO2020199144A1 WO 2020199144 A1 WO2020199144 A1 WO 2020199144A1 CN 2019081167 W CN2019081167 W CN 2019081167W WO 2020199144 A1 WO2020199144 A1 WO 2020199144A1
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receptors
alzheimer
receptor
chemokine
disease
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李翔
冯博
陈瑜
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Shenzhen Institute of Advanced Technology of CAS
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
    • C07K14/715—Receptors; Cell surface antigens; Cell surface determinants for cytokines; for lymphokines; for interferons

Definitions

  • the invention belongs to the technical field of biomedicine, and specifically relates to a therapeutic target for Alzheimer's disease.
  • AD Alzheimer's Disease
  • AD is a degenerative central nervous system degenerative disease with insidious onset and progressive development, characterized by progressive memory loss and dementia.
  • AD has become the fourth killer of human beings after heart, cerebrovascular disease, and cancer, and it seriously affects more and more elderly people.
  • AD International Federation (Alzheimer’s Association) survey report shows that the total number of AD patients worldwide currently exceeds 35 million, and it is expected that this number will increase to 115 million by 2050.
  • my country’s sixth national census in 2010, China’s population of 60 years and older has reached 178 million, accounting for 13.3% of the country’s total population. It is estimated that by 2050, the aging population will reach 430 million. How to detect and diagnose the disease early is the key to effectively reduce the occurrence of AD.
  • the pathological features of AD include: First, the appearance of ⁇ -amyloid protein outside the neuron cell ( ⁇ -amyloid peptide, A ⁇ ) senile plaques formed by excessive aggregation ( senile plaques, SP); the second is neurofibrillary tangles (neurofibrillary tangles) formed by the aggregation of highly phosphorylated TAU protein in neuronal cells. tangle, NFT). As the most important component of age spots, A ⁇ plays an extremely important role in the occurrence and development of Alzheimer's.
  • a ⁇ is a normal product of a variety of cells. Under pathological conditions, ⁇ -amyloid precursor protein is hydrolyzed by ⁇ - and ⁇ -secretase to produce A ⁇ peptide fragments of 40 amino acid residues or 42 amino acid residues.
  • the oligomers formed by A ⁇ 42 can directly produce toxic effects on neurons, cause mitochondrial damage, induce neuronal apoptosis, accelerate the appearance of neurofibrillary tangles, and further cause spatial memory loss.
  • a ⁇ activates astrocytes and microglia by acting on multiple receptors on blood monocytes or glial cells in the brain, and the activated astrocytes and microglia produce a variety of inflammatory reactions Complement, inflammatory cytokines, induce inflammation in the brain and aggravate the deterioration of AD patients. Therefore, the discovery of A ⁇ 42-related functional receptors is of great significance to promote the development of AD research. Studies have reported that there are several endogenous A ⁇ receptors on microglia, astrocytes or neurons, such as scavenger receptor (SR), receptor for glycosylation end products (RAGE), and G protein coupling Receptor (FPR2) and so on.
  • SR scavenger receptor
  • RAGE receptor for glycosylation end products
  • FPR2 G protein coupling Receptor
  • the purpose of the present invention is to overcome the above-mentioned shortcomings of the prior art and provide a specific receptor for ⁇ -amyloid oligomers, a therapeutic target for Alzheimer's disease.
  • a specific receptor for ⁇ -amyloid oligomers a therapeutic target for Alzheimer's disease.
  • a ⁇ receptors especially A ⁇ oligomers.
  • one aspect of the present invention provides a specific receptor for ⁇ -amyloid oligomer.
  • the specific receptor of the ⁇ -amyloid oligomer is at least one chemokine receptor including CC receptors, CXC receptors, C receptors and CX3C receptors.
  • the CC receptor includes at least one of CCR2, CCR3, CCR4, and CCR1.
  • the CXC receptor includes at least one of CXCR1 and CXCR2.
  • the class C receptor includes XCR1.
  • the CX3C receptor includes CX3CR1.
  • the specific receptor includes at least one of CCR2, CCR3, CCR4, and CX3CR1 chemokine receptor.
  • the Alzheimer's disease treatment target is at least one chemokine receptor including CC receptors, CXC receptors, C receptors and CX3C receptors.
  • the specific receptor includes at least one of CCR2, CCR3, CCR4, and CX3CR1 chemokine receptor.
  • the specific receptor of the ⁇ -amyloid oligomer of the present invention uses at least one chemokine including CC receptors, CXC receptors, C receptors and CX3C receptors Receptor.
  • This type of chemokine receptor can specifically bind to A ⁇ oligomers, and is activated by A ⁇ oligomers to initiate downstream signaling pathways.
  • the Alzheimer’s disease treatment target of the present invention utilizes at least one chemokine receptor including CC receptors, CXC receptors, C receptors and CX3C receptors to have specific binding properties to A ⁇ oligomers, At least one chemokine receptor including CC receptors, CXC receptors, C receptors and CX3C receptors is used as a therapeutic target for Alzheimer's disease, which can significantly improve Alzheimer's disease star Symptoms such as glial cell activation and spatial memory impairment.
  • Figure 1 is a schematic flow chart of a method for screening A ⁇ oligomer-specific receptors using a cell model in an embodiment
  • Figure 2 is a histogram of the related performance of using specific chemokine receptors as targets to improve the activation of astrocytes in Alzheimer’s transgenic mice in Example 2.
  • Figure 2-A is a wild-type mouse The morphology of astrocytes in the hippocampus of transgenic mice with Alzheimer's disease
  • Figure 2-B shows the percentage of astrocyte area
  • Figure 2-C shows wild-type mice and Alzheimer's transgenic mice The movement distance of the mice in a certain period of time in the mine experiment.
  • Figure 2-D shows the movement time of wild-type mice and Alzheimer's transgenic mice in the starting arm and other arms in the Y maze experiment.
  • Figure 2-E shows the shuttle probability of wild-type mice and Alzheimer's transgenic mice in the Y maze experiment in the starting arm.
  • AD Alzheimer’s Disease
  • ⁇ -amyloid peptide, A ⁇ is composed of amyloid precursor protein (amyloid precursor protein) protein, APP) A polypeptide containing 39 to 43 amino acids produced by the proteolysis of ⁇ - and ⁇ -secretase.
  • a ⁇ oligomer It is formed by the aggregation of A ⁇ protein and spontaneous conversion, including low molecular weight oligomers below 10 mers and high molecular weight oligomers with a degree of polymerization of 20-40. It and fibrous aggregates are the main pathological changes in the brain of AD patients.
  • CC receptors CC-chemokine Receptor (CCR).
  • CXC receptors CXC chemokine receptor (CXCR).
  • Class C receptor C chemokines receptor (CR).
  • CX3C receptors CX3C chemokine receptor (CX3CR).
  • CCR2 CC-chemokine Receptor2 (CCR2).
  • CCR3 CC-chemokine Receptor3 (CCR3).
  • CCR4 CC-chemokine Receptor4 (CCR4).
  • CX3CR1 is a specific receptor for Fractalkine, belonging to the chemokine receptor superfamily, with 7 transmembrane G-protein coupling domains. Its gene is located at 3p21-3pter, close to the CCR gene family. CX3CR1 is mainly expressed on cytotoxic effector lymphocytes (including NK cells, cytotoxic T lymphocytes and macrophages). It is a highly selective chemokine receptor and surface marker on cytotoxic effector lymphocytes. The cells highly express cell particles such as perforin and granzyme B.
  • Chemokine receptors belong to a superfamily of G protein-coupled receptors that span 7 times. They are usually expressed on cell membranes such as immune cells and endothelial cells. After binding to chemokines, they activate downstream effectors to participate in inflammation. Based on a large number of research findings, the inventors of the present invention specifically screened A ⁇ oligomer-specific receptors according to the cell model shown in Figure 1 and found that A ⁇ oligomers are used as ligands, which can interact with CC receptors and CXC receptors. At least one chemokine receptor among receptor-like receptors, C receptors and CX3C receptors specifically binds.
  • chemokine receptors After these chemokine receptors are combined with A ⁇ oligomer ligands, they activate different downstream signaling pathways to produce multiple biological functions. Its main biological functions include causing chemotaxis, generating calcium flow, and regulating inflammation, thereby causing adverse stimulation to the central nervous system.
  • inventions of the present invention provide specific receptors for ⁇ -amyloid oligomers.
  • the specific receptor of the ⁇ -amyloid oligomer is at least one chemokine receptor including CC receptors, CXC receptors, C receptors and CX3C receptors.
  • the CC receptor includes at least one of CCR1, CCR2, CCR3, and CCR4, the CXC receptor includes at least one of CXCR1 and CXCR2, and the C receptor includes XCR1
  • the CX3C receptor includes CX3CR1.
  • the specific receptor of the ⁇ -amyloid oligomer includes at least one of CCR2, CCR3, CCR4, and CX3CR1 chemokine receptor.
  • chemokine receptors specifically bind to ⁇ -amyloid oligomers, and are activated by A ⁇ oligomers to initiate downstream signaling pathways, thereby generating a variety of biological functions, such as causing chemotaxis and generating calcium flux. Regulate inflammation, as shown in Figure 1.
  • chemokine receptors on immune cells such as macrophages and dendritic cells (the common denominator of these different types of cells is the expression of chemokine receptors)
  • the activation of calcium flow signaling pathways causes internal The release of calcium simultaneously activates NF-kB and Erk
  • Different signaling pathways such as macrophages promote the migration of immune cells such as macrophages, and regulate the inflammatory response in the body, resulting in adverse effects on the central nervous system. Therefore, the above-mentioned chemokine receptors, especially at least one of the above-mentioned preferred chemokine receptors including CCR2, CCR3, CCR4, and CX3CR1
  • CCR2, CCR3, CCR4, and CX3CR1 can be used as specific receptors for A ⁇ oligomers. Based on the above-mentioned specific binding characteristics of chemokine receptors and A ⁇ oligomers, it provides a new therapeutic direction for improving and treating Alzheimer’s disease, as well as a new method for
  • the embodiments of the present invention provide new targets for the treatment of Alzheimer's disease.
  • the Alzheimer's disease treatment target is at least one chemokine receptor including CC receptors, CXC receptors, C receptors and CX3C receptors.
  • these types of chemokine receptors can be used as new targets for the treatment of Alzheimer’s disease, which can effectively prevent these types of chemokine receptors from specifically binding with A ⁇ oligomers, thereby avoiding being affected by A ⁇ oligomers.
  • the CC receptors, CXC receptors, C receptors and CX3C receptors are respectively the CC receptors, CXC receptors, C receptors and CX3C receptors described above.
  • the chemokine receptor is at least one of CCR2, CCR3, CCR4, and CX3CR1.
  • Example 1 Using cell models to screen models of A ⁇ oligomer-specific receptors (experimental methods):
  • mouse primary microglia were cultured, and A ⁇ oligomers were added to the culture medium to observe the movement tendency of microglia in real time. Knock out the CCR2, CCR3, CCR4 or CX3CR1 receptors in primary microglia, and observe the movement tendency of microglia after adding A ⁇ oligomers.
  • a confocal microscope was used to dynamically observe the morphological changes and trajectories of microglia in real time.
  • the morphology of microglia changed: the cell body became larger, the protrusions retracted, and moved to the position where the A ⁇ oligomer was added. After knocking out the CCR2, CCR3, CCR4 or CX3CR1 receptor, the microglia no longer move towards the position where A ⁇ oligomers are added.
  • Example 2 Effect experiment of chemokine receptor as a therapeutic target for Alzheimer's disease:
  • Figure 2-A shows the morphology of astrocytes in the hippocampus of wild-type mice and Alzheimer's transgenic mice
  • Figure 2-B shows the star The percentage of glial cell area
  • Figure 2-C shows the movement distance of wild-type mice and Alzheimer’s transgenic mice in a certain period of time in the mine experiment.
  • Figure 2-D shows the Y maze experiment. The activity time of wild-type mice and Alzheimer's transgenic mice in the starting arm and other arms.
  • Figure 2-E shows wild-type mice and Alzheimer's transgenic mice in the Y maze experiment. The shuttle probability of the beginning arm.
  • the astrocytes in the hippocampus of the 5XFAD transgenic mice are activated, showing that the cell body becomes larger, the protrusions retract, and the area of astrocytes increases. After knocking out the chemokine receptor, the 5XFAD transgenic mice returned to a resting state.
  • 5XFAD transgenic mice have a shorter movement distance than wild-type mice in a certain period of time in the mine experiment, and the movement distance of 5XFAD transgenic mice after chemokine receptor knockout is equivalent to that of wild-type mice.
  • the residence time of 5XFAD transgenic mice in the starting arm and the number of shuttles in the starting arm were shortened, which was manifested by impaired working memory.
  • the residence time of 5XFAD transgenic mice in the starting arm and the number of shuttles in the starting arm are equivalent to those of wild-type mice.
  • chemokine receptors as targets for the treatment of Alzheimer’s disease can effectively prevent the specific binding of these chemokine receptors to A ⁇ oligomers, thereby avoiding or reducing the initiation of downstream signals
  • the pathway produces a variety of biological functions that are unfavorable to the central nervous system, which can significantly improve the symptoms of Alzheimer’s astrocyte activation and spatial memory impairment, so as to effectively improve and treat AD.

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Abstract

β-淀粉样蛋白寡聚体的特异性受体和阿尔兹海默症治疗靶点。所述β-淀粉样蛋白寡聚体的特异性受体和阿尔兹海默症治疗靶点为包括CC类受体、CXC类受体、C受体和CX3C受体中的至少一种趋化因子受体。该类型的趋化因子受体能够特异性与Aβ寡聚体结合，并被Aβ寡聚体激活后启动下游信号通路，以该类型的趋化因子受体作为阿尔兹海默症治疗靶点，可显著性改善阿尔兹海默症星形胶质细胞激活及空间记忆损伤等症状。

Description

阿尔兹海默症治疗靶点 技术领域
本发明属于生物医药技术领域，具体的是涉及一种阿尔兹海默症治疗靶点。
 背景技术
阿尔茨海默病(Alzheimer’s Disease，AD)是一种起病隐匿的进行性发展的中枢神经系统退行性疾病，以进行性记忆缺失和痴呆为临床特征。AD已成为继心、脑血管病、癌症后的人类第四号杀手，其严重影响着越来越多的老年人。根据2010年阿尔茨海默病国际联合会（Alzheimer’s Association）调查报告显示，目前全球AD患者总数已经超过3500万，预计到2050年该数目将增至1.15亿。我国2010年第六次全国人口普查数据显示，中国60岁及以上人口已经达到1.78亿，占全国总人口的13.3%，预计到2050年，老龄化人口将达到4.3亿。如何能够早期发现和诊断该疾病，是有效降低AD发生的关键。
AD病理特征包括：一是神经元细胞外出现以β-淀粉样蛋白 (β-amyloid peptide，Aβ)过度聚集形成的老年斑( senile plaques, SP)；二是神经元胞体内出现高度磷酸化TAU 蛋白聚集所形成的神经纤维缠结(neurofibrillary tangle, NFT)。Aβ作为老年斑最为重要的成分，在阿尔茨海默的发生发展中发挥着极为重要的作用。
Aβ是多种细胞的正常产物。在病理状态下，β淀粉样前体蛋白被β-和γ-分泌酶水解产生40个氨基酸残基或42个氨基酸残基的Aβ肽段。Aβ42 形成的寡聚体可直接对神经元产生毒性作用，引起线粒体损伤，诱导神经元凋亡，加速神经纤维缠结的出现，并进一步造成空间记忆缺失。Aβ 通过作用于血液单核细胞或者脑内胶质细胞上的多种受体，激活星型胶质细胞与小胶质细胞，活化的星型胶质细胞与小胶质细胞产生多种炎症反应补体，炎症细胞因子，诱发脑内炎症反应，加重AD 病人的恶化。因此发现Aβ42 相关的功能性受体对推动AD研究发展具有重要意义。研究报道小胶质细胞，星型胶质细胞或神经元上存在数种内源性Aβ受体，如清道夫受体(SR)、糖基化终产物受体(RAGE)、G蛋白偶联受体（FPR2）等。但是这些受体能否作为检测Aβ42寡聚体依旧存在争议，而且临床已发现的这些受体特异性不高，并没有特异性针对有神经毒性的Aβ寡聚体。因此，基于已发现的受体治疗不能改善阿尔兹海默症的多种症状。
 技术问题
本发明的目的在于克服现有技术的上述不足，提供一种β-淀粉样蛋白寡聚体的特异性受体，阿尔兹海默症治疗靶点。以解决目前针对Aβ受体特别是Aβ寡聚体存在争议，特异性不高而导致的技术问题。
 技术解决方案
为了实现上述发明目的，本发明的一方面，提供一种β-淀粉样蛋白寡聚体的特异性受体。所述β-淀粉样蛋白寡聚体的特异性受体为包括CC类受体、CXC类受体、C受体和CX3C受体中的至少一种趋化因子受体。
优选地，所述CC类受体包括CCR2、CCR3、CCR4、CCR1中的至少一种。
优选地，所述CXC类受体包括CXCR1、CXCR2中的至少一种。
优选地，优选地，所述C类受体包括XCR1。
优选地，所述CX3C类受体包括CX3CR1。
优选地，所述特异性受体为包括CCR2、CCR3、CCR4、CX3CR1趋化因子受体中的至少一种。
本发明的另一方面，提供了一种阿尔兹海默症治疗靶点。所述阿尔兹海默症治疗靶点为包括CC类受体、CXC类受体、C受体和CX3C受体中的至少一种趋化因子受体。
优选地，所述特异性受体为包括CCR2、CCR3、CCR4、CX3CR1趋化因子受体中的至少一种。
 有益效果
与现有技术相比，本发明β-淀粉样蛋白寡聚体的特异性受体是采用包括CC类受体、CXC类受体、C受体和CX3C受体中的至少一种趋化因子受体。该类型的趋化因子受体能够特异性与Aβ寡聚体结合，并被Aβ寡聚体激活后启动下游信号通路。
本发明阿尔兹海默症治疗靶点利用包括CC类受体、CXC类受体、C受体和CX3C受体中的至少一种趋化因子受体具有与Aβ寡聚体特异性结合特性，将包括CC类受体、CXC类受体、C受体和CX3C受体中的至少一种趋化因子受体作为阿尔兹海默症治疗靶点，可显著性改善阿尔兹海默症星形胶质细胞激活及空间记忆损伤等症状。
 附图说明
下面将结合附图及实施例对本发明作进一步说明，附图中：
图1为实施例采用细胞模型筛选Aβ寡聚体特异性受体的方法流程示意图；
图2为实施例2中以特异性趋化因子受体为靶点改善阿尔兹海默症转基因小鼠星形胶质细胞激活的相关性能柱状图；其中，图2-A为野生型小鼠和阿尔兹海默症转基因小鼠海马区域星形胶质细胞形态，图2-B为星形胶质细胞面积所占的百分比，图2-C为野生型小鼠和阿尔兹海默症转基因小鼠在矿场实验中一定时间内运动的距离，图2-D为在Y迷宫实验中，野生型小鼠和阿尔兹海默症转基因小鼠在起始臂和其他臂中的活动时间，图2-E为野生型小鼠和阿尔兹海默症转基因小鼠在Y迷宫实验中在起始臂的穿梭几率。
 本发明的最佳实施方式
为了使本发明要解决的技术问题、技术方案及有益效果更加清楚明白，以下结合实施例与附表，对本发明进行进一步详细说明。应当理解，此处所描述的具体实施例仅仅用以解释本发明，并不用于限定本发明。
本文中涉及的相关专用名称的解释：
阿尔茨海默病(Alzheimer’s Disease，AD) ：是一种起病隐匿的进行性发展的中枢神经系统退行性疾病，以进行性记忆缺失和痴呆为临床特征，又名老年痴呆。
β淀粉样蛋白(β-amyloid peptide，Aβ)：是由淀粉样前体蛋白(amyloid precursor protein，APP) 经 β-和γ-分泌酶的蛋白水解作用而产生的含有39～43个氨基酸的多肽。
Aβ寡聚体：是由Aβ蛋白聚集而自发转换形成的，包括10聚以下的低分子量的寡聚体和聚合度为20-40的高分子量的寡聚体。其与纤维状聚集物都是AD患者脑内主要的病理改变。
CC类受体：CC-chemokine Receptor (CCR)。
CXC类受体：CXC chemokine receptor (CXCR)。
C类受体：C chemokines receptor (CR)。
CX3C类受体：CX3C chemokine receptor (CX3CR)。
CCR2：CC趋化因子受体2（CC-chemokine Receptor2 (CCR2)）。
CCR3：CC趋化因子受体3（CC-chemokine Receptor3 (CCR3)）。
CCR4：CC趋化因子受体4（CC-chemokine Receptor4 (CCR4)）。
CX3CR1：CX3CR1是Fractalkine的特异性受体，属于趋化因子受体超家族，具有7次跨膜G-蛋白偶联结构域。其基因定位于3p21 - 3pter ，临近CCR基因族。CX3CR1主要在细胞毒性效应淋巴细胞（包括NK细胞、细胞毒T淋巴细胞和巨噬细胞）上表达，是细胞毒性效应淋巴细胞上一种高选择性的趋化因子受体和表面标志，而这些细胞高表达穿孔素和颗粒酶B等细胞颗粒。
趋化因子受体属于7次跨膜的G蛋白耦联受体超家族，通常表达于免疫细胞、内皮细胞等细胞膜上，与趋化因子结合后活化下游效应物参与炎症反应。本发明的发明人基于大量研究发现，具体的按照图1所示的细胞模型筛选Aβ寡聚体特异性受体的方法发现Aβ寡聚体作为配体，其能够与包括CC类受体、CXC类受体、C受体和CX3C受体中的至少一种趋化因子受体特异性结合。该些趋化因子受体与Aβ寡聚体配体结合后，通过激活下游不同的信号通路，产生多种生物学功能。其主要的生物学功能包括引起趋化，产生钙流，调节炎症反应，从而造成对中枢神经系统不良刺激。
基于此，本发明实施例提供β-淀粉样蛋白寡聚体的特异性受体。所述β-淀粉样蛋白寡聚体的特异性受体为包括CC类受体、CXC类受体、C类受体和CX3C类受体中的至少一种趋化因子受体。
在具体实施例中，所述CC类受体包括CCR1、CCR2、CCR3、CCR4中的至少一种，所述CXC类受体包括CXCR1、CXCR2中的至少一种，所述C类受体包括XCR1，所述CX3C类受体包括CX3CR1。在优选实施例中，所述所述β-淀粉样蛋白寡聚体的特异性受体包括CCR2、CCR3、CCR4、CX3CR1趋化因子受体中的至少一种。
上述趋化因子受体与β-淀粉样蛋白寡聚体特异性结合，并被Aβ寡聚体激活后启动下游信号通路，从而产生多种生物学功能，如包括引起趋化，产生钙流，调节炎症反应，具体的如图1所示。具体如免疫细胞如巨噬细胞，树突细胞（这些不同类型的细胞的共同点是表达有趋化因子受体）上的趋化因子受体被激活后，通过激活钙流信号通路，引起内钙的释放，同时激活NF-kB，Erk 等不同的信号通路，促进巨噬细胞等免疫细胞迁移，调节体内炎症反应，从而导致对中枢神经系统的不利作用。因此，上文所述的趋化因子受体，特别是上文优选的包括CCR2、CCR3、CCR4、CX3CR1中的至少一种趋化因子受体能够作为Aβ寡聚体的特异性受体。基于上文所述的趋化因子受体与Aβ寡聚体特异性结合特性，为改善和治疗阿尔兹海默病提供了新的治疗方向，同时也为检测Aβ寡聚体提供了一种新的方法。
基于上文所述的趋化因子受体与Aβ寡聚体之间具有的特异性结合关系，本发明实施例提供了阿尔兹海默症治疗新靶点。所述阿尔兹海默症治疗靶点为包括CC类受体、CXC类受体、C受体和CX3C受体中的至少一种趋化因子受体。这样将该些类的趋化因子受体作为阿尔兹海默症治疗新靶点，能够有效避免该些类趋化因子受体与Aβ寡聚体发生特异性结合，从而避免被Aβ寡聚体激活后启动下游信号通路而产生多种对中枢神经系统不利的如图1所示的生物学功能，以显著性改善阿尔兹海默症星形胶质细胞激活及空间记忆损伤等症状，达到有效改善和治疗AD的效果。在具体实施例中，所述CC类受体、CXC类受体、C受体和CX3C受体分别如上文中所述的CC类受体、CXC类受体、C受体和CX3C受体，在优选实施例中，所述趋化因子受体为CCR2、CCR3、CCR4、CX3CR1中的至少一种。
现结合具体实例，对本发明进行进一步详细说明。
实施例1. 采用细胞模型筛选Aβ寡聚体特异性受体的模型（实验方法）：
（1）实验方法如下：
参照图1所示的筛选Aβ寡聚体特异性受体的模型，培养小鼠原代小胶质细胞，在培养基中加入Aβ寡聚体，实时观察小胶质细胞的运动趋向。分别敲除原代小胶质细胞中的CCR2、CCR3、CCR4或者CX3CR1受体，观察加入Aβ寡聚体后小胶质细胞的运动趋向。
（2）实验检测方法：
采用共聚焦显微镜实时动态观察小胶质细胞的形态变化及运动轨迹。
（3）实验结果：
培养基中加入Aβ寡聚体后，小胶质细胞形态发生改变：细胞胞体变大，突起回缩，并向Aβ寡聚体加入的位置移动。敲除CCR2、CCR3、CCR4或者CX3CR1受体后，小胶质细胞不再趋向Aβ寡聚体加入的位置移动。
实施例2. 趋化因子受体为阿尔兹海默症治疗靶点的效果实验：
（1）实验方法如下：
1）免疫化学实验：取野生型小鼠、5XFAD转基因小鼠及趋化因子受体敲除的5XFAD转基因小鼠的大脑海马组织进行冰冻切片，使用星形胶质细胞特异性抗体进行免疫组织化学染色，观察星形胶质细胞的形态；
2）行为学实验：选取矿场实验、Y迷宫行为学范式，观察野生型小鼠、5XFAD转基因小鼠及趋化因子受体敲除的5XFAD转基因小鼠在上述行为学范式中的运动能力及工作记忆能力。
（2）实验检测方法：
1）免疫化学检测：采用星形胶质细胞特异性抗体GFAP对不同小鼠的海马冰冻切片进行免疫化染色，采用共聚焦显微镜观察星形胶质细胞的形态：细胞胞体大小，突起长短。
2）行为学检测：矿场实验中，记录不同小鼠在矿场实验中一定时间内的运动距离。Y迷宫中，记录不同小鼠在Y迷宫起始臂和其他臂中停留的时间，以及在起始臂中穿梭的次数占在各个臂中穿梭次数的百分比。
（3）实验结果：实验结果如图2所示，其中，图2-A为野生型小鼠和阿尔兹海默症转基因小鼠海马区域星形胶质细胞形态，图2-B为星形胶质细胞面积所占的百分比，图2-C为野生型小鼠和阿尔兹海默症转基因小鼠在矿场实验中一定时间内运动的距离，图2-D为在Y迷宫实验中，野生型小鼠和阿尔兹海默症转基因小鼠在起始臂和其他臂中的活动时间，图2-E为野生型小鼠和阿尔兹海默症转基因小鼠在Y迷宫实验中在起始臂的穿梭几率。
1）5XFAD转基因小鼠海马中星形胶质细胞为激活状态，表现为细胞胞体变大，突起回缩，星形胶质细胞的面积占比增多。趋化因子受体敲除后5XFAD转基因小鼠恢复至静息状态。
2）5XFAD转基因小鼠在矿场实验中一定时间内运动距离较野生型小鼠变短，趋化因子受体敲除后5XFAD转基因小鼠的运动距离与野生型小鼠相当。在Y迷宫实验中，5XFAD转基因小鼠在起始臂中停留时间以及在起始臂穿梭次数变短，表现为工作记忆受损。趋化因子受体敲除后5XFAD转基因小鼠在起始臂中的停留时间以及在起始臂穿梭次数与野生型小鼠相当。
结论：干预Aβ寡聚体的特异性受体能够改善5XFAD转基因小鼠的病理改变及行为学损伤。
结合上述研究实验结果可知，CC类受体、CXC类受体、C受体和CX3C受体中的至少一种趋化因子受体，具体为包括CCR2、CCR3、CCR4、CX3CR1趋化因子受体中的至少一种能够与β-淀粉样蛋白寡聚体特异性结合，结合后启动下游信号通路，从而产生对中枢神经系统不利的多种生物学功能。基于此，将该些类趋化因子受体作为阿尔兹海默症治疗靶点，能够有效避免该些类趋化因子受体与Aβ寡聚体发生特异性结合，从而避免或降低启动下游信号通路而产生多种对中枢神经系统不利的生物学功能，以显著性改善阿尔兹海默症星形胶质细胞激活及空间记忆损伤等症状，达到有效改善和治疗AD的效果。
以上所述的实施例仅为本发明的较佳实施例而已，并不用以限制本发明，凡在本发明的精神和原则之内所作的任何修改、等同替换和改进等，均应包含在本发明的保护范围之内。

Claims (8)

  1. β-淀粉样蛋白寡聚体的特异性受体，其特征在于：所述特异性受体为包括CC类受体、CXC类受体、C类受体和CX3C类受体中的至少一种趋化因子受体。
  2. 根据权利要求1所述的特异性受体，其特征在于：所述CC类受体包括CCR2、CCR3、CCR4、CCR1中的至少一种。
  3. 根据权利要求1所述的特异性受体，其特征在于：所述CXC类受体包括CXCR1、CXCR2中的至少一种。
  4. 根据权利要求1所述的特异性受体，其特征在于：所述C类受体包括XCR1。
  5. 根据权利要求1所述的特异性受体，其特征在于：所述CX3C类受体包括CX3CR1。
  6. 根据权利要求1所述的特异性受体，其特征在于：所述特异性受体为包括CCR2、CCR3、CCR4、CX3CR1趋化因子受体中的至少一种。
  7. 阿尔兹海默症治疗靶点，其特征在于：所述阿尔兹海默症治疗靶点为包括CC类受体、CXC类受体、C受体和CX3C受体中的至少一种趋化因子受体。
  8. 根据权利要求7所述的阿尔兹海默症治疗靶点，其特征在于：所述特异性受体为包括CCR2、CCR3、CCR4、CX3CR1趋化因子受体中的至少一种。
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MENG QI XIA, ET AL.: "Chemokines/chemokine receptors in the central nervous system and Alzheimer's disease", JOURNAL OF NEUROVIROLOGY, vol. 5, 31 December 1999 (1999-12-31), XP055193965, DOI: 20190509124706X *

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