CN107050013A - 一种褐藻多酚用于制备治疗阿尔茨海默病的药物的用途 - Google Patents

一种褐藻多酚用于制备治疗阿尔茨海默病的药物的用途 Download PDF

Info

Publication number
CN107050013A
CN107050013A CN201710011185.XA CN201710011185A CN107050013A CN 107050013 A CN107050013 A CN 107050013A CN 201710011185 A CN201710011185 A CN 201710011185A CN 107050013 A CN107050013 A CN 107050013A
Authority
CN
China
Prior art keywords
rwe
brown algae
medicine
purposes
algae polyphenols
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Granted
Application number
CN201710011185.XA
Other languages
English (en)
Other versions
CN107050013B (zh
Inventor
崔巍
何山
王钦文
陈海敏
张金荣
严小军
王嘉玲
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Ningbo University
Original Assignee
Ningbo University
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Ningbo University filed Critical Ningbo University
Priority to CN201710011185.XA priority Critical patent/CN107050013B/zh
Publication of CN107050013A publication Critical patent/CN107050013A/zh
Application granted granted Critical
Publication of CN107050013B publication Critical patent/CN107050013B/zh
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/357—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having two or more oxygen atoms in the same ring, e.g. crown ethers, guanadrel
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/02—Algae
    • A61K36/03—Phaeophycota or phaeophyta (brown algae), e.g. Fucus

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Epidemiology (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

本发明公开了下式的褐藻多酚用于制备治疗阿尔茨海默病的药物的用途。该褐藻多酚有极强的抗氧化作用，对Aβ纤维化和寡聚化具有抑制作用，有抗AD神经保护活性，安全性好。

Description

一种褐藻多酚用于制备治疗阿尔茨海默病的药物的用途
技术领域
本发明涉及一种褐藻多酚(RWE)的用途，尤其是涉及该褐藻多酚用于制备治疗阿尔茨海默病的药物的用途。
背景技术
褐藻多酚即RWE是从褐藻门翅藻科昆布属的腔昆布Ecklonia cava中提取的一种多酚类化合物，具有极强的抗氧化性，可有效清除自由基和单线态氧，预防自由基损伤。研究发现，腔昆布Ecklonia cava具有多种食用和药用价值，在人体中可有效对抗氧化应激，具有抗肿瘤、消炎抗菌、抗过敏等多种活性。
阿尔兹海默病(Alzheimer’s diseases,AD)是一种以β淀粉样蛋白(β-amyloid，Aβ)聚集为主要特征的神经退行性疾病。目前，AD的发病机制仍不明确，存在多种学说，如Aβ沉积说、tau蛋白理论、炎性损伤说、金属离子中毒说、氧化应激学说等。研究表明，异常沉积Aβ诱导的氧化应激导致的神经元代谢障碍和突触功能丧失在AD发病中处于关键地位。氧化应激不仅在AD早期起作用，还激活信号转导通路引起级联酶促反应，推动AD病程进展。过多的活性氧自由基还可诱导β淀粉样蛋白聚集和tau蛋白的磷酸化。因此，降低氧化应激水平，可防止或减少Aβ在脑内聚集，是防治AD的重要靶点和治疗策略。
目前应用的抗氧化剂包括生理性、天然和化学合成三类。天然抗氧化剂主要有黄酮类、异黄酮类、多酚类、芪类化合物、姜黄素、烟碱等，由于其易于摄取且副作用少，并且在自然界大量存在而备受关注。
发明内容
本发明提供一种具有如下结构的褐藻多酚用于制备治疗阿尔茨海默病的药物的用途：
与现有技术相比，本发明的优点在于：褐藻多酚RWE可以有效清除活性氧自由基(ROS)生成，并且具有安全性好的优势，可多靶点协同对抗AD。同时，研究表明褐藻多酚RWE能够有效地抑制Aβ寡聚化、纤维化，并能降低神经毒性，安全性好，适合作为治疗阿尔茨海默病的药物。
附图说明
图1为体外检测RWE对自由基的清除能力；
图2为体外检测RWE对Aβ纤维化的阻止能力；
图3为体外检测RWE对Aβ寡聚化的阻止能力；
图4为细胞活性实验；
图5为2',7'-二氯荧光黄双乙酸盐(DCF-DA)细胞染色实验；
图6为荧光素双醋酸酯(FDA)/碘化丙啶(PI)细胞染色实验
图7为Hoechst细胞染色；
图8为动物旷场试验5分钟内站立次数；
图9为动物旷场试验5分钟内跨线次数。
具体实施方式
以下结合附图和实施例对本发明作进一步详细描述。
实施例一
RWE从腔昆布中提取纯化获得，其化学结构式如下：
本发明所涉及化合物RWE片剂的制备：
取5克化合物RWE加入糊精195克，混匀，常规压片制成1000片。
实施例二
本发明所涉及化合物RWE胶囊剂的制备：
取5克化合物RWE加入淀粉195克，混匀，装胶囊制成1000粒。
实施例三
体外检测RWE对自由基的清除能力。DPPH(1,1-Diphenyl-2-picrylhydrazylradical)即1,1-二苯基-2-苦基肼基自由基购自sigma公司，溶于甲醇中配成1mg/ml溶液，避光保存，现配现用。96孔板中，设置0.1-100μMRWE为实验组，30μM维生素C(VitC)为阳性对照组，每孔加入药物10μl，再加入90μl的DPPH溶液，避光室温反应15min，酶标仪419nm下读数。RWE加入后读数降低(图1)，说明RWE能够在体外清除自由基。
实施例四
体外检测RWE对Aβ纤维形成的阻碍能力。Aβ粉末(Sigma公司)在六氟异丙醇(HFIP)中溶解，至终浓度2.5mg/ml。储存于-80℃冰箱中待用，使用时将100μl Aβ溶液挥发，加入60μl 20mM氢氧化钠溶液，氮吹15分钟。检测Aβ纤维化时，10μM Aβ与0.1-10μM的RWE以及5μM硫磺素-T混合，37℃孵育3天，使用荧光酶标仪读数(激发波长440nm，发射波长485nm)。RWE加入后酶标仪读数降低(图2)，说明RWE能够体外降低Aβ纤维生成，显示RWE可对AD过程中的毒性Aβ纤维形成有阻止作用。
实施例五
体外检测RWE对Aβ寡聚体形成的阻碍能力。Aβ粉末在六氟异丙醇中溶解，至终浓度2.5mg/ml。储存于-80℃冰箱中待用，使用时加入900μl双蒸水，氮吹15分钟。离心15000rpm15分钟后，取上清。震荡48小时后即获得60μM的Aβ寡聚体(Aβoligomers)。震荡前，加入10μMRWE，震荡48小时。RWE处理与未处理的Aβ寡聚体经醋酸铀染色后，透射电镜观察。RWE处理组Aβ未明显形成寡聚体(图3)，说明RWE能够体外降低Aβ寡聚体生成，显示RWE可对AD过程中的毒性Aβ寡聚体形成有阻止作用。
实施例六
检测RWE对Aβ寡聚体细胞毒性的阻止能力。SH-SY5Y细胞(来源于中国科学院上海生命科学研究院)，加入如前述1.5μMAβ寡聚体的细胞为模型组。提前0.5小时加入1-20μM的RWE为实验组，用MTT比色法检测细胞存活率。由图5可知，SH-SY5Y细胞中加入Aβ寡聚体，可导致细胞死亡，提前0.5小时加入RWE，可降低Aβ寡聚体导致的细胞死亡率，得出结论RWE可对抗Aβ寡聚体造成的细胞毒性，有神经保护作用。由于Aβ可通过在细胞内生成双氧水，产生氧化应激导致细胞死亡，这从一个角度说明RWE可对抗氧化应激产生神经保护。
DCF-DA购自sigma公司,溶于二甲亚砜(DMSO)中配成100mM即10mg/0.188ml DCF-DA储存液，避光-20℃保存，使用时用DMEM(Dulbecco minimum essential medium)培养基稀释至合适配比。
对照组为未加处理的SH-SY5Y细胞，加入1.5μM Aβ寡聚体的细胞为模型组，提前0.5小时加入20μM RWE为实验组。用DMEM换液后，加入10μM DCF-DA溶液，37℃孵育5分钟，磷酸缓冲液润洗，留少量磷酸缓冲液于荧光显微镜下拍摄。DCF-DA染色显示绿色荧光，且绿色荧光越深表明ROS生成越多。从图5可以看到RWE可以有效降低Aβ寡聚体诱导的细胞内ROS生成，能够对抗氧化应激导致的细胞死亡。
DCF-DA(carboxy-2’,7’-dichiorodihydroflurescein diacetate)即2',7'-二氯荧光黄双乙酸盐购自sigma公司,溶于二甲亚砜(DMSO)中配成100mM即10mg/0.188ml DCF-DA储存液，避光-20℃保存，使用时用DMEM(Dulbecco minimum essential medium)培养基稀释至合适配比。
FDA和PI购自Sigma公司。两者溶于磷酸缓冲液(FDA 10μg/ml，PI 5μg/ml),配置为FDA/PI工作液。对照组为未加处理的SH-SY5Y细胞，加入1.5μM Aβ寡聚体的细胞为模型组，提前0.5小时加入20μMRWE为实验组。用DMEM换液后，加入FDA/PI工作液，37℃孵育5分钟，磷酸缓冲液润洗，留少量磷酸缓冲液于荧光显微镜下拍摄。FDA染色显示活细胞，PI染色显示死细胞。从图6中可以看到RWE可以对抗Aβ寡聚体造成的细胞毒性，有神经保护作用。
Hoechst 33342购自Sigma公司，Hoechst 33342 5μg/ml溶于磷酸缓冲液。对照组为未加处理的SH-SY5Y细胞，加入1.5μM Aβ寡聚体的细胞为模型组，提前0.5小时加入20μMRWE为实验组。用DMEM换液后，加入Hoechst 33342溶液，37℃孵育10分钟，磷酸缓冲液润洗，留少量磷酸缓冲液于荧光显微镜下拍摄。Hoechst染色显示细胞核，较亮较小的点显示凋亡的细胞核。从图7中可以看到RWE可以对抗Aβ寡聚体造成的细胞凋亡。
综上所述，RWE能够抑制Aβ寡聚体的细胞毒性，并能降低细胞内氧化应激水平，有抗AD神经保护作用。
实施例七
图8、9中动物为ICR小鼠(来源于浙江省动物中心)，腹腔注射5、10mg/kgRWE溶液为实验组，注射溶液后1小时后旷场实验检测5分钟内动物站立次数和跨线次数。由图8可知，RWE 5mg/kg和10mg/kg注射一小时后，动物站立次数不发生显著改变，由图9可知，动物跨线次数不发生显著改变。说明RWE有很好的安全性。
上述说明并非对本发明的限制，本发明也并不限于上述举例。本技术领域的普通技术人员在本发明的实质范围内，作出的变化、改型、添加或替换，也应属于本发明的保护范围。

Claims (1)

1.下式的褐藻多酚用于制备治疗阿尔茨海默病的药物的用途
CN201710011185.XA 2017-01-06 2017-01-06 一种褐藻多酚用于制备治疗阿尔茨海默病的药物的用途 Active CN107050013B (zh)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201710011185.XA CN107050013B (zh) 2017-01-06 2017-01-06 一种褐藻多酚用于制备治疗阿尔茨海默病的药物的用途

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201710011185.XA CN107050013B (zh) 2017-01-06 2017-01-06 一种褐藻多酚用于制备治疗阿尔茨海默病的药物的用途

Publications (2)

Publication Number Publication Date
CN107050013A true CN107050013A (zh) 2017-08-18
CN107050013B CN107050013B (zh) 2020-03-03

Family

ID=59624094

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201710011185.XA Active CN107050013B (zh) 2017-01-06 2017-01-06 一种褐藻多酚用于制备治疗阿尔茨海默病的药物的用途

Country Status (1)

Country Link
CN (1) CN107050013B (zh)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108434178A (zh) * 2018-04-13 2018-08-24 桂林医学院 褐藻多酚在制备防治帕金森病药物中的应用
CN115813957A (zh) * 2022-11-17 2023-03-21 江苏海洋大学 海蒿子多酚在制备治疗阿尔兹海默病药物中的应用

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2013049639A (ja) * 2011-08-30 2013-03-14 Fisheries Research Agency クロメ由来のフロロタンニン類を有効成分とする紫外線照射障害保護剤

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2013049639A (ja) * 2011-08-30 2013-03-14 Fisheries Research Agency クロメ由来のフロロタンニン類を有効成分とする紫外線照射障害保護剤

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
BYOUNG WOOK CHOI ET AL: "Multifunctional Activity of Polyphenolic Compounds Associated with a Potential for Alzheimer’s Disease Therapy from Ecklonia cava", 《PHYTOTHER. RES》 *
IL-JUN KANG ET AL: "Butanol extract of Ecklonia cava prevents production and aggregation", 《FOOD AND CHEMICAL TOXICOLOGY》 *

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108434178A (zh) * 2018-04-13 2018-08-24 桂林医学院 褐藻多酚在制备防治帕金森病药物中的应用
CN115813957A (zh) * 2022-11-17 2023-03-21 江苏海洋大学 海蒿子多酚在制备治疗阿尔兹海默病药物中的应用
CN115813957B (zh) * 2022-11-17 2023-10-27 江苏海洋大学 海蒿子多酚在制备治疗阿尔兹海默病药物中的应用

Also Published As

Publication number Publication date
CN107050013B (zh) 2020-03-03

Similar Documents

Publication Publication Date Title
Kumar et al. Antioxidant and antipyretic properties of methanolic extract of Amaranthus spinosus leaves
Wang et al. Gallic acid induces apoptosis and enhances the anticancer effects of cisplatin in human small cell lung cancer H446 cell line via the ROS-dependent mitochondrial apoptotic pathway
Zhang et al. Tetramethylpyrazine scavenges superoxide anion and decreases nitric oxide production in human polymorphonuclear leukocytes
Scholz et al. Cardiovascular ion channels as a molecular target of flavonoids
Punithavathi et al. Pretreatment with a combination of quercetin and α-tocopherol ameliorates adenosine triphosphatases and lysosomal enzymes in myocardial infarcted rats
Liang et al. Ganoderma lucidum polysaccharides target a Fas/caspase dependent pathway to induce apoptosis in human colon cancer cells
Tang et al. Houttuynia cordata Thunb extract induces apoptosis through mitochondrial-dependent pathway in HT-29 human colon adenocarcinoma cells
CN101869299B (zh) 栽培菊苣提取物及其应用
CN107050013A (zh) 一种褐藻多酚用于制备治疗阿尔茨海默病的药物的用途
Li et al. Myricetin exhibits anti-glioma potential by inducing mitochondrial-mediated apoptosis, cell cycle arrest, inhibition of cell migration and ROS generation
Jit et al. Phytochemicals: A potential next generation agent for radioprotection
Namchaiw et al. The leaf extract of Coccinia grandis (L.) Voigt accelerated in vitro wound healing by reducing oxidative stress injury
CN104958288A (zh) 一种血小板抑制剂及其在制备抗血小板疾病药物中的应用
Zhao et al. Reactive oxygen species-responsive theranostic nanoparticles for enhanced hypoxic tumor photodynamic therapy via synchronous HIF-1α inhibition and ATP depletion
Oliveira et al. Reestablishment of Redox Homeostasis in the Nociceptive Primary Afferent as a Mechanism of Antinociception Promoted by Mesenchymal Stem/Stromal Cells in Oxaliplatin‐Induced Chronic Peripheral Neuropathy
Zhou et al. FV‐429 Induced Apoptosis Through ROS‐Mediated ERK2 Nuclear Translocation and p53 Activation in Gastric Cancer Cells
Yun et al. Isoquercitrin, isolated from Aster yomena triggers ROS-mediated apoptosis in Candida albicans
Shahnazi et al. Protective effects of methanolic extract of Vicia cracca against hypoxia-induced lethality in mice
Tabrizi Fabrication of folic acid-conjugated chitosan-coated PLGA nanoparticles for targeted delivery of Peganum harmala smoke extract to breast cancer cells
Tian et al. A H2S‐Generated Supramolecular Photosensitizer for Enhanced Photodynamic Antibacterial Infection and Relieving Inflammation
Rashid et al. Membrane stabilizing activity-a possible mechanism of action for the anti-inflammatory activity of two Bangladeshi medicinal plants: Mesua nagassarium (Burm. F.) and Kigelia pinnata (Jack) Dc
CN102219772A (zh) 具有生物活性的土槿皮酸复合物及其制备方法
Lin et al. The ethanolic extract of Taiwanofungus camphoratus (Antrodia camphorata) induces cell cycle arrest and enhances cytotoxicity of cisplatin and doxorubicin on human hepatocellular carcinoma cells
Qi et al. Visnagin inhibits cervical cancer cells proliferation through the induction of apoptosis and modulation of PI3K/AKT/mTOR and MAPK signaling pathway
CN103040854A (zh) 獐牙菜苦苷及其苷元的医药用途

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant
EE01 Entry into force of recordation of patent licensing contract
EE01 Entry into force of recordation of patent licensing contract

Application publication date: 20170818

Assignee: Ningbo Science and Technology Innovation Association

Assignor: Ningbo University

Contract record no.: X2023980033633

Denomination of invention: Use of a brown algae polyphenol in the preparation of drugs for the treatment of Alzheimer's disease

Granted publication date: 20200303

License type: Common License

Record date: 20230317