WO2017085844A1 - Β-secretase inhibitor containing turmerone as active ingredient, and pharmaceutical preparation and food/drink containing said inhibitor - Google Patents
Β-secretase inhibitor containing turmerone as active ingredient, and pharmaceutical preparation and food/drink containing said inhibitor Download PDFInfo
- Publication number
- WO2017085844A1 WO2017085844A1 PCT/JP2015/082611 JP2015082611W WO2017085844A1 WO 2017085844 A1 WO2017085844 A1 WO 2017085844A1 JP 2015082611 W JP2015082611 W JP 2015082611W WO 2017085844 A1 WO2017085844 A1 WO 2017085844A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- turmerone
- secretase
- inhibitor
- secretase inhibitor
- food
- Prior art date
Links
Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
-
- 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
- A61K31/12—Ketones
Definitions
- the present invention relates to a ⁇ -secretase inhibitor containing a rhizome-derived compound of turmeric (Curcuma longa) as an active ingredient, and a pharmaceutical preparation and food and drink containing the inhibitor.
- a ⁇ -secretase inhibitor containing a rhizome-derived compound of turmeric (Curcuma longa) as an active ingredient, and a pharmaceutical preparation and food and drink containing the inhibitor.
- Alzheimer's disease a typical disease of senile dementia, is a degenerative disease characterized by brain atrophy, deposition of senile plaques, and formation of neurofibrillary tangles. Dementia symptoms are caused by neuronal loss. Is considered to be caused (Non-Patent Document 1).
- ⁇ -amyloid which is the main component of senile plaques, has a cytotoxic action and is considered to cause neuronal cell death in Alzheimer's disease (non-patent documents 2 to 5).
- ⁇ -amyloid is produced from the amyloid precursor protein (APP) by the action of an enzyme called ⁇ -secretase. Therefore, compounds having an inhibitory action on this ⁇ -secretase are considered to be useful for the prevention of Alzheimer's disease.
- Patent Document 1 research on compounds having an inhibitory action on ⁇ -secretase has been actively conducted.
- Patent Document 2 As an example of the search for a highly safe ⁇ -secretase inhibitor, the invention described in Patent Document 2 by the present inventors can be mentioned.
- ⁇ -secretase inhibitors are extracted from natural products generally used as spices such as pepper, sesame seeds and turmeric.
- Patent Document 2 further specifies a compound having ⁇ -secretase inhibitory activity among compounds contained in the extract.
- ⁇ -secretase inhibitors that have higher inhibitory activity and are cheaper.
- Patent Document 3 describes a method of treating a patient suffering from Alzheimer's disease using an extract from Spice. Specifically, the use of curcuminoid, turmeric, polysaccharides and / or turmeric extract containing turmerin is described. However, although Patent Document 3 describes that turmeric extract inhibits amyloid aggregation, it has a problem that the specific action mechanism has not been clarified.
- Patent Document 4 describes an acetylcholinesterase inhibitor containing an AR turmerone derivative to be used for the treatment of Alzheimer's disease.
- a substance having acetylcholinesterase inhibitory activity is useful for symptomatic treatment for alleviating the accumulation of ⁇ -amyloid, but has a problem that it is not effective for preventing Alzheimer's disease.
- JP 2008-137914 A JP, 2013-189385, A Japanese Patent Application Publication No. 2009-530305 JP, 2011-225478, A
- the present inventors in Japanese Patent Application No. 2015-146208, sesamolin extracted from sesame, galangal and hijasu respectively, ethyl 4-methoxycinnamate and pipetalin
- the present invention proposes a pharmaceutical preparation and food and drink having a ⁇ -secretase inhibitor containing as an active ingredient.
- the invention described in Japanese Patent Application No. 2015-146208 specifies the above three compounds as compounds having ⁇ -secretase inhibitory activity, and shows specific inhibitory activity by calculating the IC 50 value of each compound.
- ⁇ -secretase inhibitors that have higher inhibitory activity and are cheaper.
- the present invention provides a ⁇ -secretase inhibitor which is useful for the prevention of Alzheimer's disease, is highly safe, and has a further high inhibitory activity on ⁇ -secretase activity, and a pharmaceutical preparation and food and drink containing the inhibitor. It is.
- the invention according to claim 1 relates to a ⁇ -secretase inhibitor containing, as an active ingredient, one or more compounds selected from the group consisting of AR-turmerone, ⁇ -turmerone and ⁇ -turmerone.
- the invention according to claim 2 relates to the ⁇ -secretase inhibitor according to claim 1, characterized in that AR-turmerone, ⁇ -turmerone and ⁇ -turmerone are extracted from turmeric rhizomes.
- the invention according to claim 3 relates to a pharmaceutical preparation comprising the ⁇ -secretase inhibitor according to claim 1 or 2.
- the invention according to claim 4 relates to a food or drink containing the ⁇ -secretase inhibitor according to claim 1 or 2.
- the first aspect of the present invention since it has a still higher inhibitory activity on ⁇ -secretase activity, it is possible to prevent or stop the progression of senile dementia such as Alzheimer's disease.
- the compound obtained from the extract of the rhizome of turmeric (Curcuma longa) is used as an active ingredient, the safety to the human body is high, and senile dementia such as Alzheimer's disease, etc. It is possible to provide ⁇ -secretase inhibitors that can prevent or halt the progression. Furthermore, since the method of extracting a compound from turmeric is simple, it can be manufactured at low cost.
- the pharmaceutical preparation contains a ⁇ -secretase inhibitor having an excellent inhibitory action on ⁇ -secretase activity
- the ⁇ -secretase inhibitor is ingested in accordance with the condition and condition of the user. It is possible to prevent or stop the progression of senile dementia such as Alzheimer's disease.
- the ⁇ -secretase inhibitor since it is a food and drink containing a ⁇ -secretase inhibitor having an excellent inhibitory effect on ⁇ -secretase activity, the ⁇ -secretase inhibitor can be easily taken on a daily basis. And prevent the progression of senile dementia such as Alzheimer's disease.
- ⁇ -secretase inhibitor according to the present invention, and the pharmaceutical preparation and food and drink containing the inhibitor will be described.
- the ⁇ -secretase inhibitor of the present invention contains, as an active ingredient, one or more compounds selected from the group consisting of AR-turmerone, ⁇ -turmerone and ⁇ -turmerone.
- AR-turmerone is CAS number 532-65-0, and has a structure shown in the following formula (Formula 1).
- ⁇ -turmerone is CAS number 82508-15-4 and has a structure shown in the following formula (Formula 2).
- ⁇ -turmerone is CAS No. 82508-14-3 and has a structure shown in the following formula (Formula 3).
- Turmeric (English name: Turmeric, Scientific name: Curcuma longa) is a monocotyledonous plant of the ginger family Turmeric genus. It is a perennial herb native to India and having a developed rhizome, which is used as a raw material and dye for curry powder.
- AR-turmerone, ⁇ -turmerone and ⁇ -turmerone are preferably extracted from the rhizome of turmeric with hexane. The reason is that, by extracting the turmeric rhizome with hexane, it is possible to elute more AR-turmerone, ⁇ -turmerone and ⁇ -turmerone.
- the above compounds may be used singly or in combination of two or more. As described in detail in the following examples, since the above compounds have strong ⁇ -secretase inhibitory activity, the ⁇ -secretase inhibitors of the present invention containing them as an active ingredient can be used for various uses. it can.
- the ⁇ -secretase inhibitor of the present invention when used as a pharmaceutical preparation, it is used as a preventive agent for senile dementia in mammals (particularly human), particularly as a prophylactic agent for Alzheimer's disease.
- the beta-secretase inhibitor of the present invention can be tablet, powder, fine granule, granule, coated tablet, capsule, syrup, troche, inhalant, suppository, injection, ointment according to a conventional method. It can be formulated into pharmaceutical preparations such as eye ointments, eye drops, nasal drops, ear drops, buffs, lotions and the like.
- Excipients binders, lubricants, colorants, flavoring agents, and, if necessary, stabilizers, emulsifiers, absorption accelerators, surfactants, pH adjusters, preservatives, anti-oxidants which are usually used for formulation.
- An oxidizing agent and the like can be used, and in general, components and amounts to be used as raw materials of pharmaceutical preparations are appropriately selected and formulated by a standard method.
- the form thereof is not particularly limited, and it may be a commonly used method, and may be orally or parenterally administered.
- the dose of the pharmaceutical preparation according to the present invention can be appropriately selected as a pharmaceutically effective amount according to the degree of symptoms, age, sex, body weight, administration form, specific type of disease and the like.
- oral administration for example, in the case of oral administration, it is usually appropriate for an active ingredient amount of about 0.001 to 1000 mg / kg in adults, and if it is divided into once a day several times Good.
- the beta-secretase inhibitor of the present invention is a food additive such as various beverages such as soft drinks, dairy products (processed milk, yogurt), confectionery products (jelly, chocolate, biscuit, gum, tablets) or supplements. It can also be blended with various beverages such as soft drinks, dairy products (processed milk, yogurt), confectionery products (jelly, chocolate, biscuit, gum, tablets) or supplements. It can also be blended with various beverages such as soft drinks, dairy products (processed milk, yogurt), confectionery products (jelly, chocolate, biscuit, gum, tablets) or supplements. It can also be blended with various beverages such as soft drinks, dairy products (processed milk, yogurt), confectionery products (jelly, chocolate, biscuit, gum, tablets) or supplements. It can also be blended with various beverages such as soft drinks, dairy products (processed milk, yogurt), confectionery products (jelly, chocolate, biscuit, gum, tablets) or supplements. It can also be blended with various beverages such as soft drinks, dairy products (processed milk, yogurt), confectionery products (jelly, chocolate, biscuit, gum,
- the addition amount thereof is not particularly limited, and may be appropriately determined according to the type of food.
- the dry weight of the above-mentioned extract may be added so that the content is in the range of about 0.0005 to 50% by weight.
- the food and drink contain, in addition to the ⁇ -secretase inhibitor of the present invention, excipients, flavoring agents, coloring agents, preservatives, thickeners, stabilizers, gelling agents, antioxidants, etc. It is also good.
- the excipients include, but are not limited to, for example, powders such as fine particle silicon dioxide, sucrose fatty acid ester, crystalline cellulose, sodium carboxymethylcellulose, calcium hydrogen phosphate, wheat starch, rice starch, corn starch , Potato starch, dextrin, cyclodextrin and other starches, crystalline cellulose, lactose, glucose, sugar, reduced maltose, starch syrup, fructooligosaccharide, galactooligosaccharide, soybean oligosaccharide, isomaltooligosaccharide, xylooligosaccharide, maltoligosaccharide, milk and fruit Examples thereof include saccharides such as oligosaccharides, and sugar
- the flavoring agent examples include, but are not limited to, for example, bontan extract which is a fruit juice extract, lychee extract, apple juice, orange juice, orange juice, yuzu extract, peach flavor, ume flavor, acesulfame K which is a sweetening agent, erythritol, oligosaccharides And mannose, xylitol, isomerized saccharides, green tea which is a tea component, oolong tea, banaba tea, tonka tea, iron kannon tea, castorine tea, blowfish tea, amacha tea, macomo tea, kelp tea, yoghurt flavor and the like.
- bontan extract which is a fruit juice extract, lychee extract, apple juice, orange juice, orange juice, yuzu extract, peach flavor, ume flavor, acesulfame K which is a sweetening agent, erythritol, oligosaccharides And mannose, xylit
- ⁇ -secretase inhibition rate (%) 100 ⁇ [(area value of peak from each extract / peak area value from control) ⁇ 100]
- the hexane extract was analyzed using GCMS under the following conditions. ⁇ Column; Intertcap Pure Wax (0.25 id ⁇ 0.25 ⁇ m ⁇ 60 m) Injection volume: 1.0 ⁇ l -Temperature rising; holding at 50 ° C for 2 minutes, 2.5 ° C heating / minute to 240 ° C-Vaporization room temperature: 250 ° C ⁇ Carry gas flow rate: Helium 1.2 ml / min ⁇ Split ratio: 1/80 From the results of GCMS analysis, the compound was estimated to be AR-turmerone, ⁇ -turmerone and ⁇ -turmerone by attached library search (wiley7n and NIST08). The hexane extract contained 9.9% AR-turmerone, 30.4% ⁇ -turmerone, and 11.5% ⁇ -turmerone.
- the hexane extract was fractionated under the following HPLC conditions. ⁇ Column; L-column ODS (20 id ⁇ 250 mm) ⁇ Mobile phase: water: acetonitrile (20: 80, v / v) ⁇ Flow rate: 18.9 ml / min ⁇ Column temperature; Room temperature ⁇ Detection; UV 243 nm ⁇ Injection volume: 10 ⁇ l As active ingredients of the hexane extract, colorless oily substances were obtained from the fractions of RT 7.5, 10.5 and 11.0 respectively.
- ⁇ -turmerone and ⁇ -turmerone have been known as substances which are difficult to separate, but it should be noted that they could be extracted by such a simple method as described above.
- the enzyme inhibitory activity of the standard products of the above-identified compounds was examined in the same manner as above (measurement of ⁇ -secretase inhibition rate), and each standard product was examined.
- IC50 value it was as follows.
- the above-mentioned IC50 value showed that AR-turmerone, ⁇ -turmerone and ⁇ -turmerone have ⁇ -secretase inhibitory activity.
- This IC50 value is the IC50 value of sesamoline, pipetalin and ethyl 4-methoxycinnamate which the inventors previously found and described in Japanese Patent Application No. 2015-146208 (sesamoline: 0.14 mM, ethyl 4-methoxycinnamate: 0 AR-turmerone, ⁇ -turmerone and ⁇ -turmerone were found to have very high ⁇ -secretase inhibitory activity, which is significantly lower compared to 676 mM and pipetalin: 0.304 mM). Further, since the extraction method of the present invention is simple as compared with the extraction method described in Japanese Patent Application No. 2015-140208, the production cost can be suppressed to 1/10 or less.
- the present invention can be suitably used for a pharmaceutical preparation for preventing Alzheimer's disease, and can be suitably used for additives of various foods and drinks such as soft drinks, dairy products, confectioneries or supplements. it can.
Abstract
[Problem] To provide a pharmaceutical preparation and food/drink containing a β-secretase inhibitor that is highly safe and that exhibits a better inhibitory effect on β-secretase activity. [Solution] Provided is a β-secretase inhibitor that contains one or more compounds selected from a group composed of AR-turmerone, α-turmerone, and β-turmerone as active ingredients.
Description
本発明は、ターメリック（Ｃｕｒｃｕｍａ ｌｏｎｇａ）の根茎由来の化合物を有効成分として含有するβ‐セクレターゼ阻害剤、並びに該阻害剤を含む医薬製剤及び飲食品に関する。
The present invention relates to a β-secretase inhibitor containing a rhizome-derived compound of turmeric (Curcuma longa) as an active ingredient, and a pharmaceutical preparation and food and drink containing the inhibitor.
近年の急速な高齢化社会の到来に伴い、老人性痴呆症は、医学的、社会的にも重大な問題となっており、それにより有効な抗認知症治療薬の開発だけではなく、老人性痴呆症を効果的に予防できる薬の開発が強く望まれている。老人性痴呆症の代表的な疾患であるアルツハイマー型痴呆症（アルツハイマー病）は、脳の萎縮、老人班の沈着及び神経原繊維の形成を特徴とする変性疾患で、神経細胞の脱落によって痴呆症状が引き起こされると考えられている（非特許文献１）。
With the advent of the rapidly aging society in recent years, senile dementia has become a serious medical and social problem, and it is not only the development of effective anti-dementia drugs, but also senile disease. There is a strong demand for the development of drugs that can effectively prevent dementia. Alzheimer's disease (Alzheimer's disease), a typical disease of senile dementia, is a degenerative disease characterized by brain atrophy, deposition of senile plaques, and formation of neurofibrillary tangles. Dementia symptoms are caused by neuronal loss. Is considered to be caused (Non-Patent Document 1).
アルツハイマー型痴呆症の原因について未だ定説はないが、病理組織学的研究により、老人班が沈着しそれにより神経細胞が脱落し脳の萎縮が生じると考えられている。老人班の主成分であるβ‐アミロイドは細胞毒性作用を有しており、アルツハイマー型痴呆症における神経細胞死を引き起こしていると考えられている（非特許文献２～５）。β‐アミロイドは、アミロイド前駆体タンパク質（ＡＰＰ）からβ‐セクレターゼという酵素の作用によって生成される。従って、このβ‐セクレターゼに対し阻害作用を有する化合物は、アルツハイマー型痴呆症の予防に有用であると考えられる。β‐セクレターゼに対し阻害作用を有する化合物に関する研究が、近年活発に行われている（特許文献１）。
Although the cause of Alzheimer's disease has not been determined as yet, it is thought by histopathological studies that senile plaques are deposited, which leads to loss of nerve cells and atrophy of the brain. Β-amyloid, which is the main component of senile plaques, has a cytotoxic action and is considered to cause neuronal cell death in Alzheimer's disease (non-patent documents 2 to 5). β-amyloid is produced from the amyloid precursor protein (APP) by the action of an enzyme called β-secretase. Therefore, compounds having an inhibitory action on this β-secretase are considered to be useful for the prevention of Alzheimer's disease. Recently, research on compounds having an inhibitory action on β-secretase has been actively conducted (Patent Document 1).
このようなβ‐セクレターゼに対して阻害活性を持つ化合物は、β‐アミロイドの生成のみならず、他の生体内での反応をも阻害する可能性がある。他の生体内での反応を阻害するような物質は、β‐アミロイドの生成を阻害したとしても、患者に深刻な副作用をもたらす虞があるため、アルツハイマー型痴呆症等の治療に利用することは困難である。よって、アルツハイマー型痴呆症の治療に使用することができる、安全性の高いβ‐セクレターゼ阻害剤が求められている。
Compounds having inhibitory activity against such β-secretase may inhibit not only the formation of β-amyloid but also other in vivo reactions. Other substances that inhibit the reaction in the living body may cause serious side effects to patients even if they inhibit the formation of β-amyloid, so it may be used for treatment of Alzheimer's disease, etc. Have difficulty. Thus, there is a need for a highly safe β-secretase inhibitor that can be used for the treatment of Alzheimer's disease.
安全性の高いβ－セクレターゼ阻害剤を探索した例として、本発明者らによる特許文献２記載の発明が挙げられる。
特許文献２では、安全性の高いβ－セクレターゼ阻害剤を得るために、例えばコショウ、ゴマ、ターメリック等のスパイスとして一般的に用いられる天然物からβ－セクレターゼ阻害剤が抽出されている。特許文献２ではさらに抽出物に含まれる化合物の内β－セクレターゼ阻害活性を示す化合物が特定されている。
しかし、より阻害活性が高く且つより安価で得られるβ－セクレターゼ阻害剤が求められている。 As an example of the search for a highly safe β-secretase inhibitor, the invention described in Patent Document 2 by the present inventors can be mentioned.
In Patent Document 2, in order to obtain a highly safe β-secretase inhibitor, β-secretase inhibitors are extracted from natural products generally used as spices such as pepper, sesame seeds and turmeric. Patent Document 2 further specifies a compound having β-secretase inhibitory activity among compounds contained in the extract.
However, there is a need for β-secretase inhibitors that have higher inhibitory activity and are cheaper.
特許文献２では、安全性の高いβ－セクレターゼ阻害剤を得るために、例えばコショウ、ゴマ、ターメリック等のスパイスとして一般的に用いられる天然物からβ－セクレターゼ阻害剤が抽出されている。特許文献２ではさらに抽出物に含まれる化合物の内β－セクレターゼ阻害活性を示す化合物が特定されている。
しかし、より阻害活性が高く且つより安価で得られるβ－セクレターゼ阻害剤が求められている。 As an example of the search for a highly safe β-secretase inhibitor, the invention described in Patent Document 2 by the present inventors can be mentioned.
In Patent Document 2, in order to obtain a highly safe β-secretase inhibitor, β-secretase inhibitors are extracted from natural products generally used as spices such as pepper, sesame seeds and turmeric. Patent Document 2 further specifies a compound having β-secretase inhibitory activity among compounds contained in the extract.
However, there is a need for β-secretase inhibitors that have higher inhibitory activity and are cheaper.
特許文献３は、スパイスからの抽出物を利用してアルツハイマー型痴呆症を罹患している患者を処置する方法について記載している。具体的には、クルクミノイド、ターメロン、多糖類及び／又はターメリンを含むウコン抽出物を使用することが記載されている。
しかし、特許文献３はウコン抽出物がアミロイド凝集を阻害することが記載されているものの、具体的な作用機序が明らかにされていないという問題点を有する。 Patent Document 3 describes a method of treating a patient suffering from Alzheimer's disease using an extract from Spice. Specifically, the use of curcuminoid, turmeric, polysaccharides and / or turmeric extract containing turmerin is described.
However, although Patent Document 3 describes that turmeric extract inhibits amyloid aggregation, it has a problem that the specific action mechanism has not been clarified.
しかし、特許文献３はウコン抽出物がアミロイド凝集を阻害することが記載されているものの、具体的な作用機序が明らかにされていないという問題点を有する。 Patent Document 3 describes a method of treating a patient suffering from Alzheimer's disease using an extract from Spice. Specifically, the use of curcuminoid, turmeric, polysaccharides and / or turmeric extract containing turmerin is described.
However, although Patent Document 3 describes that turmeric extract inhibits amyloid aggregation, it has a problem that the specific action mechanism has not been clarified.
特許文献４には、アルツハイマー型痴呆症の治療に用いるＡＲターメロン誘導体を含むアセチルコリンエステラーゼ阻害剤が記載されている。
しかし、アセチルコリンエステラーゼ阻害活性を有する物質はβ－アミロイドの蓄積を緩和する対症療法に役立つが、アルツハイマー型痴呆症の予防に効果的であるとはいえないという問題点を有する。 Patent Document 4 describes an acetylcholinesterase inhibitor containing an AR turmerone derivative to be used for the treatment of Alzheimer's disease.
However, a substance having acetylcholinesterase inhibitory activity is useful for symptomatic treatment for alleviating the accumulation of β-amyloid, but has a problem that it is not effective for preventing Alzheimer's disease.
しかし、アセチルコリンエステラーゼ阻害活性を有する物質はβ－アミロイドの蓄積を緩和する対症療法に役立つが、アルツハイマー型痴呆症の予防に効果的であるとはいえないという問題点を有する。 Patent Document 4 describes an acetylcholinesterase inhibitor containing an AR turmerone derivative to be used for the treatment of Alzheimer's disease.
However, a substance having acetylcholinesterase inhibitory activity is useful for symptomatic treatment for alleviating the accumulation of β-amyloid, but has a problem that it is not effective for preventing Alzheimer's disease.
安全性の高いβ－セクレターゼ阻害剤を探索したもう一つの例として、本発明者らは特願２０１５－１４６２０８において、それぞれゴマ、ガランガル及びヒハツから抽出されたセサモリン、エチル４－メトキシシンナメート及びピパタリンを有効成分として含むβ－セクレターゼ阻害剤を有する医薬製剤及び飲食品を提案している。
特願２０１５－１４６２０８記載の発明は、β－セクレターゼ阻害活性を有する化合物として上記３つの化合物を特定しており、それぞれの化合物のＩＣ５０値を算出することで具体的な阻害活性を示している。
しかし、より阻害活性が高く且つより安価で得られるβ－セクレターゼ阻害剤が求められている。 As another example of the search for a highly safe beta-secretase inhibitor, the present inventors in Japanese Patent Application No. 2015-146208, sesamolin extracted from sesame, galangal and hijasu respectively, ethyl 4-methoxycinnamate and pipetalin The present invention proposes a pharmaceutical preparation and food and drink having a β-secretase inhibitor containing as an active ingredient.
The invention described in Japanese Patent Application No. 2015-146208 specifies the above three compounds as compounds having β-secretase inhibitory activity, and shows specific inhibitory activity by calculating the IC 50 value of each compound.
However, there is a need for β-secretase inhibitors that have higher inhibitory activity and are cheaper.
特願２０１５－１４６２０８記載の発明は、β－セクレターゼ阻害活性を有する化合物として上記３つの化合物を特定しており、それぞれの化合物のＩＣ５０値を算出することで具体的な阻害活性を示している。
しかし、より阻害活性が高く且つより安価で得られるβ－セクレターゼ阻害剤が求められている。 As another example of the search for a highly safe beta-secretase inhibitor, the present inventors in Japanese Patent Application No. 2015-146208, sesamolin extracted from sesame, galangal and hijasu respectively, ethyl 4-methoxycinnamate and pipetalin The present invention proposes a pharmaceutical preparation and food and drink having a β-secretase inhibitor containing as an active ingredient.
The invention described in Japanese Patent Application No. 2015-146208 specifies the above three compounds as compounds having β-secretase inhibitory activity, and shows specific inhibitory activity by calculating the IC 50 value of each compound.
However, there is a need for β-secretase inhibitors that have higher inhibitory activity and are cheaper.
本発明は、アルツハイマー型痴呆症の予防に役立ち、安全性が高く、β‐セクレターゼ活性に対するさらに高い阻害活性を有するβ‐セクレターゼ阻害剤、並びに該阻害剤を含む医薬製剤及び飲食品を提供するものである。
The present invention provides a β-secretase inhibitor which is useful for the prevention of Alzheimer's disease, is highly safe, and has a further high inhibitory activity on β-secretase activity, and a pharmaceutical preparation and food and drink containing the inhibitor. It is.
請求項１に係る発明は、ＡＲ－ターメロン、α－ターメロン及びβ－ターメロンからなる群から選択される１以上の化合物を有効成分として含有するβ‐セクレターゼ阻害剤に関する。
The invention according to claim 1 relates to a β-secretase inhibitor containing, as an active ingredient, one or more compounds selected from the group consisting of AR-turmerone, α-turmerone and β-turmerone.
請求項２に係る発明は、ＡＲ－ターメロン、α－ターメロン及びβ－ターメロンが、ターメリックの根茎より抽出されてなることを特徴とする請求項１記載のβ‐セクレターゼ阻害剤に関する。
The invention according to claim 2 relates to the β-secretase inhibitor according to claim 1, characterized in that AR-turmerone, α-turmerone and β-turmerone are extracted from turmeric rhizomes.
請求項３に係る発明は、請求項１又は２に記載のβ‐セクレターゼ阻害剤を含む医薬製剤に関する。
The invention according to claim 3 relates to a pharmaceutical preparation comprising the β-secretase inhibitor according to claim 1 or 2.
請求項４に係る発明は、請求項１又は２に記載のβ－セクレターゼ阻害剤を含む飲食品に関する。
The invention according to claim 4 relates to a food or drink containing the β-secretase inhibitor according to claim 1 or 2.
請求項１に係る発明によれば、β‐セクレターゼ活性に対するさらに高い阻害活性を有するので、アルツハイマー型痴呆症等の老人性痴呆症を予防又は進行を停止することができる。
According to the first aspect of the present invention, since it has a still higher inhibitory activity on β-secretase activity, it is possible to prevent or stop the progression of senile dementia such as Alzheimer's disease.
請求項２に係る発明によれば、ターメリック（Ｃｕｒｃｕｍａ ｌｏｎｇａ）の根茎の抽出物から得られた化合物を有効成分とするため、人体に対する安全性が高く、アルツハイマー型痴呆症等の老人性痴呆症を予防又は進行を停止することができるβ‐セクレターゼ阻害剤を提供することができる。
さらに、ターメリックから化合物を抽出する方法が簡便であるため、低いコストで製造することができる。 According to the invention of claim 2, since the compound obtained from the extract of the rhizome of turmeric (Curcuma longa) is used as an active ingredient, the safety to the human body is high, and senile dementia such as Alzheimer's disease, etc. It is possible to provide β-secretase inhibitors that can prevent or halt the progression.
Furthermore, since the method of extracting a compound from turmeric is simple, it can be manufactured at low cost.
さらに、ターメリックから化合物を抽出する方法が簡便であるため、低いコストで製造することができる。 According to the invention of claim 2, since the compound obtained from the extract of the rhizome of turmeric (Curcuma longa) is used as an active ingredient, the safety to the human body is high, and senile dementia such as Alzheimer's disease, etc. It is possible to provide β-secretase inhibitors that can prevent or halt the progression.
Furthermore, since the method of extracting a compound from turmeric is simple, it can be manufactured at low cost.
請求項３に係る発明によれば、β‐セクレターゼ活性に対する優れた阻害作用を有するβ‐セクレターゼ阻害剤を含む医薬製剤であるため、使用者の症状や状態に合わせてβ‐セクレターゼ阻害剤を摂取することができ、アルツハイマー型痴呆症等の老人性痴呆症を予防又は進行を停止することができる。
According to the invention of claim 3, since the pharmaceutical preparation contains a β-secretase inhibitor having an excellent inhibitory action on β-secretase activity, the β-secretase inhibitor is ingested in accordance with the condition and condition of the user. It is possible to prevent or stop the progression of senile dementia such as Alzheimer's disease.
請求項４に係る発明によれば、β‐セクレターゼ活性に対する優れた阻害作用を有するβ‐セクレターゼ阻害剤を含む飲食品であるため、β‐セクレターゼ阻害剤を日常的に容易に摂取することができ、アルツハイマー型痴呆症等の老人性痴呆症を予防又は進行を停止することができる。
According to the invention of claim 4, since it is a food and drink containing a β-secretase inhibitor having an excellent inhibitory effect on β-secretase activity, the β-secretase inhibitor can be easily taken on a daily basis. And prevent the progression of senile dementia such as Alzheimer's disease.
以下、本発明に係るβ－セクレターゼ阻害剤、並びに該阻害剤を含む医薬製剤及び飲食品について説明する。
Hereinafter, the β-secretase inhibitor according to the present invention, and the pharmaceutical preparation and food and drink containing the inhibitor will be described.
本発明のβ‐セクレターゼ阻害剤は、ＡＲ－ターメロン、α－ターメロン及びβ－ターメロンからなる群から選択される１以上の化合物を有効成分として含有する。
The β-secretase inhibitor of the present invention contains, as an active ingredient, one or more compounds selected from the group consisting of AR-turmerone, α-turmerone and β-turmerone.
ＡＲ－ターメロンは、ＣＡＳ番号５３２－６５－０であって、下式（化１）に示す構造を有している。
AR-turmerone is CAS number 532-65-0, and has a structure shown in the following formula (Formula 1).
α－ターメロンは、ＣＡＳ番号８２５０８－１５－４であって、下式（化２）に示す構造を有している。
Α-turmerone is CAS number 82508-15-4 and has a structure shown in the following formula (Formula 2).
β－ターメロンは、ＣＡＳ番号８２５０８‐１４‐３であって、下式（化３）に示す構造を有している。
Β-turmerone is CAS No. 82508-14-3 and has a structure shown in the following formula (Formula 3).
本発明において、ＡＲ－ターメロン、α－ターメロン及びβターメロンは標準品を用いることができる。また、カレースパイス等の抽出物から得たものを用いることができ、特にターメリックの抽出物から得たものを用いることができる。
In the present invention, standard products of AR-turmerone, α-turmerone and β-turmerone can be used. Moreover, what was obtained from extracts, such as curry spice, can be used, and in particular, those obtained from extracts of turmeric can be used.
ターメリック（英名：Ｔｕｒｍｅｒｉｃ、学名：Ｃｕｒｃｕｍａ ｌｏｎｇａ）は、ショウガ科ウコン属の単子葉植物である。インド原産で、発達した根茎を有する多年草であり、根茎はカレー粉の原料や染料として用いられる。
Turmeric (English name: Turmeric, Scientific name: Curcuma longa) is a monocotyledonous plant of the ginger family Turmeric genus. It is a perennial herb native to India and having a developed rhizome, which is used as a raw material and dye for curry powder.
本発明のβ‐セクレターゼ阻害剤には、ＡＲターメロン、α－ターメロン及びβ－ターメロンを有効成分とするものを用いることができる。
ＡＲ－ターメロン、α－ターメロン及びβ－ターメロンはターメリックの根茎からヘキサンによって好適に抽出される。その理由は、ターメリックの根茎をヘキサンで抽出することにより、ＡＲ－ターメロン、α－ターメロン及びβターメロンを多く溶出させることができるからである。 As the β-secretase inhibitor of the present invention, those containing AR termeron, α-turmerone and β-turmerone as active ingredients can be used.
AR-turmerone, α-turmerone and β-turmerone are preferably extracted from the rhizome of turmeric with hexane. The reason is that, by extracting the turmeric rhizome with hexane, it is possible to elute more AR-turmerone, α-turmerone and β-turmerone.
ＡＲ－ターメロン、α－ターメロン及びβ－ターメロンはターメリックの根茎からヘキサンによって好適に抽出される。その理由は、ターメリックの根茎をヘキサンで抽出することにより、ＡＲ－ターメロン、α－ターメロン及びβターメロンを多く溶出させることができるからである。 As the β-secretase inhibitor of the present invention, those containing AR termeron, α-turmerone and β-turmerone as active ingredients can be used.
AR-turmerone, α-turmerone and β-turmerone are preferably extracted from the rhizome of turmeric with hexane. The reason is that, by extracting the turmeric rhizome with hexane, it is possible to elute more AR-turmerone, α-turmerone and β-turmerone.
上記化合物は、１種単独で又は２種以上を混合して使用してもよい。下記実施例で詳述するように、上記化合物は強いβ‐セクレターゼ阻害活性を有しているので、これらを有効成分として含有する本発明のβ‐セクレターゼ阻害剤は、各種用途に使用することができる。
The above compounds may be used singly or in combination of two or more. As described in detail in the following examples, since the above compounds have strong β-secretase inhibitory activity, the β-secretase inhibitors of the present invention containing them as an active ingredient can be used for various uses. it can.
例えば、本発明のβ‐セクレターゼ阻害剤を医薬製剤として用いる場合、哺乳動物（特にヒト）における老人性痴呆症の予防薬、特にアルツハイマー型痴呆症の予防薬として用いられる。
For example, when the β-secretase inhibitor of the present invention is used as a pharmaceutical preparation, it is used as a preventive agent for senile dementia in mammals (particularly human), particularly as a prophylactic agent for Alzheimer's disease.
本発明のβ‐セクレターゼ阻害剤は、慣用されている方法により錠剤、散剤、細粒剤、顆粒剤、被覆錠剤、カプセル剤、シロップ剤、トローチ剤、吸入剤、坐剤、注射剤、軟膏剤、眼軟膏剤、点眼剤、点鼻剤、点耳剤、バップ剤、ローション剤等の医薬製剤に製剤化することができる。
The beta-secretase inhibitor of the present invention can be tablet, powder, fine granule, granule, coated tablet, capsule, syrup, troche, inhalant, suppository, injection, ointment according to a conventional method. It can be formulated into pharmaceutical preparations such as eye ointments, eye drops, nasal drops, ear drops, buffs, lotions and the like.
製剤化に通常使用される賦形剤、結合剤、滑沢剤、着色剤、矯味矯臭剤、および必要により安定化剤、乳化剤、吸収促進剤、界面活性剤、ｐＨ調整剤、防腐剤、抗酸化剤などを使用することができ、一般に医薬製剤の原料として使用される成分及び配合量を適宜選択して定法により製剤化される。
Excipients, binders, lubricants, colorants, flavoring agents, and, if necessary, stabilizers, emulsifiers, absorption accelerators, surfactants, pH adjusters, preservatives, anti-oxidants which are usually used for formulation. An oxidizing agent and the like can be used, and in general, components and amounts to be used as raw materials of pharmaceutical preparations are appropriately selected and formulated by a standard method.
本発明の医薬製剤を投与する場合、その形態は特に限定されず、通常使用される方法であればよく、経口投与でも非経口投与でもよい。本発明に係る医薬製剤の投与量は、症状の程度、年齢、性別、体重、投与形態、疾患の具体的な種類等に応じて、製剤学的な有効量を適宜選択することができる。投与量の一例を挙げると、経口投与の場合、通常、成人において、有効成分量として０．００１～１０００ｍｇ／ｋｇ程度が適当であり、これを１日１回～数回に分けて投与すればよい。
When the pharmaceutical preparation of the present invention is administered, the form thereof is not particularly limited, and it may be a commonly used method, and may be orally or parenterally administered. The dose of the pharmaceutical preparation according to the present invention can be appropriately selected as a pharmaceutically effective amount according to the degree of symptoms, age, sex, body weight, administration form, specific type of disease and the like. In the case of oral administration, for example, in the case of oral administration, it is usually appropriate for an active ingredient amount of about 0.001 to 1000 mg / kg in adults, and if it is divided into once a day several times Good.
本発明のβ‐セクレターゼ阻害剤は、食品添加剤として、例えば清涼飲料水、乳製品（加工乳、ヨーグルト）、菓子類（ゼリー、チョコレート、ビスケット、ガム、錠菓）又はサプリメント等の各種飲食品に配合することもできる。
The beta-secretase inhibitor of the present invention is a food additive such as various beverages such as soft drinks, dairy products (processed milk, yogurt), confectionery products (jelly, chocolate, biscuit, gum, tablets) or supplements. It can also be blended with
食品添加剤として使用する場合、その添加量については、特に限定されず、食品の種類に応じて適宜決定すればよい。一例としては、上記した抽出物の乾燥重量として、含有量が０．０００５～５０重量％程度の範囲となるように添加すればよい。
When used as a food additive, the addition amount thereof is not particularly limited, and may be appropriately determined according to the type of food. As an example, the dry weight of the above-mentioned extract may be added so that the content is in the range of about 0.0005 to 50% by weight.
上記飲食品は、本発明のβ‐セクレターゼ阻害剤の他に、賦形剤、呈味剤、着色剤、保存剤、増粘剤、安定剤、ゲル化剤、酸化防止剤等を含有してもよい。
このうち、賦形剤としては、これらに限定されないが例えば、微粒子二酸化ケイ素のような粉末類、ショ糖脂肪酸エステル、結晶セルロース・カルボキシメチルセルロースナトリウム、リン酸水素カルシウム、小麦デンプン，米デンプン，トウモロコシデンプン，バレイショデンプン，デキストリン，シクロデキストリン等のでんぷん類、結晶セルロース類、乳糖，ブドウ糖，砂糖，還元麦芽糖，水飴，フラクトオリゴ糖，ガラクトオリゴ糖，大豆オリゴ糖，イソマルトオリゴ糖，キシロオリゴ糖，マルトオリゴ糖，乳果オリゴ糖などの糖類、ソルビトール，エリストール，キシリトール，ラクチトール，マンニトール等の糖アルコール類が挙げられる。これらの賦形剤は、単独で又は二種以上組み合わせて使用することができる。 The food and drink contain, in addition to the β-secretase inhibitor of the present invention, excipients, flavoring agents, coloring agents, preservatives, thickeners, stabilizers, gelling agents, antioxidants, etc. It is also good.
Among these, the excipients include, but are not limited to, for example, powders such as fine particle silicon dioxide, sucrose fatty acid ester, crystalline cellulose, sodium carboxymethylcellulose, calcium hydrogen phosphate, wheat starch, rice starch, corn starch , Potato starch, dextrin, cyclodextrin and other starches, crystalline cellulose, lactose, glucose, sugar, reduced maltose, starch syrup, fructooligosaccharide, galactooligosaccharide, soybean oligosaccharide, isomaltooligosaccharide, xylooligosaccharide, maltoligosaccharide, milk and fruit Examples thereof include saccharides such as oligosaccharides, and sugar alcohols such as sorbitol, erythritol, xylitol, lactitol and mannitol. These excipients can be used alone or in combination of two or more.
このうち、賦形剤としては、これらに限定されないが例えば、微粒子二酸化ケイ素のような粉末類、ショ糖脂肪酸エステル、結晶セルロース・カルボキシメチルセルロースナトリウム、リン酸水素カルシウム、小麦デンプン，米デンプン，トウモロコシデンプン，バレイショデンプン，デキストリン，シクロデキストリン等のでんぷん類、結晶セルロース類、乳糖，ブドウ糖，砂糖，還元麦芽糖，水飴，フラクトオリゴ糖，ガラクトオリゴ糖，大豆オリゴ糖，イソマルトオリゴ糖，キシロオリゴ糖，マルトオリゴ糖，乳果オリゴ糖などの糖類、ソルビトール，エリストール，キシリトール，ラクチトール，マンニトール等の糖アルコール類が挙げられる。これらの賦形剤は、単独で又は二種以上組み合わせて使用することができる。 The food and drink contain, in addition to the β-secretase inhibitor of the present invention, excipients, flavoring agents, coloring agents, preservatives, thickeners, stabilizers, gelling agents, antioxidants, etc. It is also good.
Among these, the excipients include, but are not limited to, for example, powders such as fine particle silicon dioxide, sucrose fatty acid ester, crystalline cellulose, sodium carboxymethylcellulose, calcium hydrogen phosphate, wheat starch, rice starch, corn starch , Potato starch, dextrin, cyclodextrin and other starches, crystalline cellulose, lactose, glucose, sugar, reduced maltose, starch syrup, fructooligosaccharide, galactooligosaccharide, soybean oligosaccharide, isomaltooligosaccharide, xylooligosaccharide, maltoligosaccharide, milk and fruit Examples thereof include saccharides such as oligosaccharides, and sugar alcohols such as sorbitol, erythritol, xylitol, lactitol and mannitol. These excipients can be used alone or in combination of two or more.
呈味剤としては、これらに限定されないが例えば、果汁エキスであるボンタンエキス、ライチエキス、リンゴ果汁、オレンジ果汁、ゆずエキス、ピーチフレーバー、ウメフレーバー、甘味剤であるアセスルファムＫ、エリストール、オリゴ糖類、マンノース、キシリトール、異性化糖類、茶成分である緑茶、ウーロン茶、バナバ茶、杜仲茶、鉄観音茶、ハトムギ茶、アマチャヅル茶、マコモ茶、昆布茶、及びヨーグルトフレーバー等が挙げられる。
Examples of the flavoring agent include, but are not limited to, for example, bontan extract which is a fruit juice extract, lychee extract, apple juice, orange juice, orange juice, yuzu extract, peach flavor, ume flavor, acesulfame K which is a sweetening agent, erythritol, oligosaccharides And mannose, xylitol, isomerized saccharides, green tea which is a tea component, oolong tea, banaba tea, tonka tea, iron kannon tea, castorine tea, blowfish tea, amacha tea, macomo tea, kelp tea, yoghurt flavor and the like.
以下、本発明の実施例を説明することにより、本発明の効果をより明確なものとする。但し、本発明は以下の実施例には限定されない。
Hereinafter, the effects of the present invention will be made clearer by describing examples of the present invention. However, the present invention is not limited to the following examples.
（ヘキサン抽出物の調製）
ターメリックの根茎１ｋｇを粉砕し、４０℃のヘキサン５０００ｍｌで１時間の攪拌抽出を行い、上澄み液４２００ｍｌをろ紙でろ過した。
次いで、その残渣に４０℃のヘキサン２５００ｍｌで０．５時間の攪拌抽出を行い、ろ紙でろ過した。得られた抽出液に含まれる溶媒をエバポレーター（４０℃）で留去し、ターメリックのヘキサン抽出物を得た。 (Preparation of hexane extract)
1 kg of turmeric rhizome was crushed, extraction with stirring was performed with 5000 ml of hexane at 40 ° C. for 1 hour, and 4200 ml of supernatant was filtered through filter paper.
The residue was then extracted by stirring with 500 ml of hexane at 40 ° C. for 0.5 hours and filtered through a filter paper. The solvent contained in the obtained extract was distilled off with an evaporator (40 ° C.) to obtain a hexane extract of turmeric.
ターメリックの根茎１ｋｇを粉砕し、４０℃のヘキサン５０００ｍｌで１時間の攪拌抽出を行い、上澄み液４２００ｍｌをろ紙でろ過した。
次いで、その残渣に４０℃のヘキサン２５００ｍｌで０．５時間の攪拌抽出を行い、ろ紙でろ過した。得られた抽出液に含まれる溶媒をエバポレーター（４０℃）で留去し、ターメリックのヘキサン抽出物を得た。 (Preparation of hexane extract)
1 kg of turmeric rhizome was crushed, extraction with stirring was performed with 5000 ml of hexane at 40 ° C. for 1 hour, and 4200 ml of supernatant was filtered through filter paper.
The residue was then extracted by stirring with 500 ml of hexane at 40 ° C. for 0.5 hours and filtered through a filter paper. The solvent contained in the obtained extract was distilled off with an evaporator (40 ° C.) to obtain a hexane extract of turmeric.
（酢酸エチル抽出物の調製）
ターメリックのヘキサン抽出後の残渣を乾燥させ、４０℃の酢酸エチル５０００ｍｌで１時間撹拌抽出し、上澄み液４８００ｍｌをろ紙でろ過した。得られた抽出液に含まれる溶媒をエバポレーター（４０℃）で留去し、ターメリックの酢酸エチル抽出物を得た。
ヘキサン抽出物及び酢酸エチル抽出物の収率を以下の表１に示す。 (Preparation of ethyl acetate extract)
The residue after hexane extraction of turmeric was dried, extracted by stirring with 5000 ml of ethyl acetate at 40 ° C. for 1 hour, and 4800 ml of supernatant was filtered through a filter paper. The solvent contained in the obtained extract was distilled off with an evaporator (40 ° C.) to obtain an ethyl acetate extract of turmeric.
The yields of hexane extract and ethyl acetate extract are shown in Table 1 below.
ターメリックのヘキサン抽出後の残渣を乾燥させ、４０℃の酢酸エチル５０００ｍｌで１時間撹拌抽出し、上澄み液４８００ｍｌをろ紙でろ過した。得られた抽出液に含まれる溶媒をエバポレーター（４０℃）で留去し、ターメリックの酢酸エチル抽出物を得た。
ヘキサン抽出物及び酢酸エチル抽出物の収率を以下の表１に示す。 (Preparation of ethyl acetate extract)
The residue after hexane extraction of turmeric was dried, extracted by stirring with 5000 ml of ethyl acetate at 40 ° C. for 1 hour, and 4800 ml of supernatant was filtered through a filter paper. The solvent contained in the obtained extract was distilled off with an evaporator (40 ° C.) to obtain an ethyl acetate extract of turmeric.
The yields of hexane extract and ethyl acetate extract are shown in Table 1 below.
（β‐セクレターゼ阻害率の測定）
上記のように抽出したヘキサン抽出物及び酢酸エチル抽出物の酵素阻害活性を、以下の方法により検討した。また、Lys-Thr-Glu-Glu-Ile-Ser-Glu-Val-Asn-Sta-Val-Ala-Glu-Phe(Sta: (3S,4S)-4-アミノ-3-ヒドロキシ-6-メチル-ヘプタン酸;ペプチド研究所製)を陽性対照として用いた。
具体的には、まず各抽出物及び陽性対照について、それぞれを含むＤＭＳＯ溶液２μｌを、０．０２Ｍ酢酸ナトリウム緩衝液（ｐＨ４．５、０．１％Ｔｒｉｔｏｎ Ｘ－１００）７８μｌとともに０．６ｍｌサンプルチューブに入れた。コントロールとして２μｌのＤＭＳＯを上記の７８μｌの０．０２Ｍ酢酸ナトリウム緩衝液とともに０．６ｍｌサンプルチューブに入れた。
その後、酵素溶液（β－セクレターゼ、17.4 μg protein/ml）10 μlを入れて混合し、37℃で10分間前培養した。
基質としてMOCAc-Ser-Glu-Val-Asn-Leu-Asp-Ala-Glu-Phe-Arg-Lys(Dnp)-Arg-Arg-NH2 （ペプチド研究所製）0.1 mmol/l溶液を10 μl加えて混合し、37℃で1時間培養した。反応後，2.5 M 酢酸ナトリウム溶液50 μlを加え、反応を停止させた。 (Measurement of β-secretase inhibition rate)
The enzyme inhibitory activity of the hexane extract and ethyl acetate extract extracted as described above was examined by the following method. In addition, Lys-Thr-Glu-Glu-Ile-Ser-Glu-Val-Asn-Sta-Val-Ala-Glu-Phe (Sta: (3S, 4S) -4-amino-3-hydroxy-6-methyl- Heptane acid (manufactured by Peptide Laboratories) was used as a positive control.
Specifically, first, for each extract and positive control, 2 μl of each containing DMSO solution, 0.6 ml sample tube together with 78 μl of 0.02 M sodium acetate buffer (pH 4.5, 0.1% Triton X-100) Put in. As a control, 2 μl of DMSO was placed in a 0.6 ml sample tube with 78 μl of 0.02 M sodium acetate buffer as described above.
Thereafter, 10 μl of an enzyme solution (β-secretase, 17.4 μg protein / ml) was added and mixed, and pre-incubated at 37 ° C. for 10 minutes.
Add 10 μl of 0.1 mmol / l solution of MOCAc-Ser-Glu-Val-Asn-Leu-Asp-Ala-Glu-Phe-Arg-Lys (Dnp) -Arg-Arg-NH 2 (manufactured by Peptide Laboratories) The mixture was mixed and cultured at 37 ° C. for 1 hour. After the reaction, 50 μl of 2.5 M sodium acetate solution was added to stop the reaction.
上記のように抽出したヘキサン抽出物及び酢酸エチル抽出物の酵素阻害活性を、以下の方法により検討した。また、Lys-Thr-Glu-Glu-Ile-Ser-Glu-Val-Asn-Sta-Val-Ala-Glu-Phe(Sta: (3S,4S)-4-アミノ-3-ヒドロキシ-6-メチル-ヘプタン酸;ペプチド研究所製)を陽性対照として用いた。
具体的には、まず各抽出物及び陽性対照について、それぞれを含むＤＭＳＯ溶液２μｌを、０．０２Ｍ酢酸ナトリウム緩衝液（ｐＨ４．５、０．１％Ｔｒｉｔｏｎ Ｘ－１００）７８μｌとともに０．６ｍｌサンプルチューブに入れた。コントロールとして２μｌのＤＭＳＯを上記の７８μｌの０．０２Ｍ酢酸ナトリウム緩衝液とともに０．６ｍｌサンプルチューブに入れた。
その後、酵素溶液（β－セクレターゼ、17.4 μg protein/ml）10 μlを入れて混合し、37℃で10分間前培養した。
基質としてMOCAc-Ser-Glu-Val-Asn-Leu-Asp-Ala-Glu-Phe-Arg-Lys(Dnp)-Arg-Arg-NH2 （ペプチド研究所製）0.1 mmol/l溶液を10 μl加えて混合し、37℃で1時間培養した。反応後，2.5 M 酢酸ナトリウム溶液50 μlを加え、反応を停止させた。 (Measurement of β-secretase inhibition rate)
The enzyme inhibitory activity of the hexane extract and ethyl acetate extract extracted as described above was examined by the following method. In addition, Lys-Thr-Glu-Glu-Ile-Ser-Glu-Val-Asn-Sta-Val-Ala-Glu-Phe (Sta: (3S, 4S) -4-amino-3-hydroxy-6-methyl- Heptane acid (manufactured by Peptide Laboratories) was used as a positive control.
Specifically, first, for each extract and positive control, 2 μl of each containing DMSO solution, 0.6 ml sample tube together with 78 μl of 0.02 M sodium acetate buffer (pH 4.5, 0.1% Triton X-100) Put in. As a control, 2 μl of DMSO was placed in a 0.6 ml sample tube with 78 μl of 0.02 M sodium acetate buffer as described above.
Thereafter, 10 μl of an enzyme solution (β-secretase, 17.4 μg protein / ml) was added and mixed, and pre-incubated at 37 ° C. for 10 minutes.
Add 10 μl of 0.1 mmol / l solution of MOCAc-Ser-Glu-Val-Asn-Leu-Asp-Ala-Glu-Phe-Arg-Lys (Dnp) -Arg-Arg-NH 2 (manufactured by Peptide Laboratories) The mixture was mixed and cultured at 37 ° C. for 1 hour. After the reaction, 50 μl of 2.5 M sodium acetate solution was added to stop the reaction.
反応液100 μlを、水900 μlが入ったバイアル瓶に添加し、以下の条件の蛍光HPLC分析に供した。
・カラム；L-column ODS （4.6 id × 250 mm）
・移動相；water / 0.1% formic acid : acetonitrile（9：1 v/v）→17.50 min（11：9，v/v）→17.51 min（1：19，v/v）→22.50 min（1：19，v/v）→22.51 min（9：1，v/v）→27.50 min（STOP）
・カラム温度；40℃
・検出；Ex. 325 nm / Em. 395 nm
・注入量；20 μl
生成した蛍光ペプチド断片のピーク面積値から，下記の式より阻害率（%）を算出した。
β－セクレターゼ阻害率（%）= 100－〔（各抽出物由来のピークの面積値／コントロール由来のピーク面積値）×100〕 100 μl of the reaction solution was added to a vial containing 900 μl of water, and subjected to fluorescence HPLC analysis under the following conditions.
・ Column; L-column ODS (4.6 id × 250 mm)
Mobile phase: water / 0.1% formic acid: acetonitrile (9: 1 v / v) → 17.50 min (11: 9, v / v) → 17.51 min (1:19, v / v) → 22.50 min (1: 19, v / v) → 22.51 min (9: 1, v / v) → 27.50 min (STOP)
・ Column temperature; 40 ° C
Detection: Ex. 325 nm / Em. 395 nm
· Injection volume: 20 μl
From the peak area value of the generated fluorescent peptide fragment, the inhibition rate (%) was calculated by the following equation.
β-secretase inhibition rate (%) = 100 − [(area value of peak from each extract / peak area value from control) × 100]
・カラム；L-column ODS （4.6 id × 250 mm）
・移動相；water / 0.1% formic acid : acetonitrile（9：1 v/v）→17.50 min（11：9，v/v）→17.51 min（1：19，v/v）→22.50 min（1：19，v/v）→22.51 min（9：1，v/v）→27.50 min（STOP）
・カラム温度；40℃
・検出；Ex. 325 nm / Em. 395 nm
・注入量；20 μl
生成した蛍光ペプチド断片のピーク面積値から，下記の式より阻害率（%）を算出した。
β－セクレターゼ阻害率（%）= 100－〔（各抽出物由来のピークの面積値／コントロール由来のピーク面積値）×100〕 100 μl of the reaction solution was added to a vial containing 900 μl of water, and subjected to fluorescence HPLC analysis under the following conditions.
・ Column; L-column ODS (4.6 id × 250 mm)
Mobile phase: water / 0.1% formic acid: acetonitrile (9: 1 v / v) → 17.50 min (11: 9, v / v) → 17.51 min (1:19, v / v) → 22.50 min (1: 19, v / v) → 22.51 min (9: 1, v / v) → 27.50 min (STOP)
・ Column temperature; 40 ° C
Detection: Ex. 325 nm / Em. 395 nm
· Injection volume: 20 μl
From the peak area value of the generated fluorescent peptide fragment, the inhibition rate (%) was calculated by the following equation.
β-secretase inhibition rate (%) = 100 − [(area value of peak from each extract / peak area value from control) × 100]
上記計算より各ヘキサン抽出物及び酢酸エチル抽出物のβ－セクレターゼ阻害率を算出した結果、以下の通りであった。
The β-secretase inhibition ratio of each hexane extract and ethyl acetate extract was calculated from the above calculation, and the results were as follows.
上記阻害率により、ターメリックの抽出物が、非常に高いβ‐セクレターゼ阻害活性を有することがわかった。
From the above inhibition rates, it was found that the extract of turmeric has very high β-secretase inhibitory activity.
ヘキサン抽出物を以下の条件でのＧＣＭＳを用いて分析した。
・カラム；Intertcap Pure Wax (0.25 i.d. × 0.25 μm × 60 m)
・注入量；1.0 μl
・昇温；50℃で2分保持、2.5℃昇温／分→240℃まで
・気化室温；250℃
・キャリーガス流量；ヘリウム 1.2 ml/分
・スプリット比；1/80
ＧＣＭＳ分析の結果から付属のライブラリー検索（wiley7nおよびNIST08）で化合物をＡＲ－ターメロン、α－ターメロン及びβターメロンと推定した。
ヘキサン抽出物はＡＲ－ターメロンを９．９％、α－ターメロンを３０．４％、及びβ－ターメロンを１１．５％含有していた。 The hexane extract was analyzed using GCMS under the following conditions.
・ Column; Intertcap Pure Wax (0.25 id × 0.25 μm × 60 m)
Injection volume: 1.0 μl
-Temperature rising; holding at 50 ° C for 2 minutes, 2.5 ° C heating / minute to 240 ° C-Vaporization room temperature: 250 ° C
・ Carry gas flow rate: Helium 1.2 ml / min ・ Split ratio: 1/80
From the results of GCMS analysis, the compound was estimated to be AR-turmerone, α-turmerone and β-turmerone by attached library search (wiley7n and NIST08).
The hexane extract contained 9.9% AR-turmerone, 30.4% α-turmerone, and 11.5% β-turmerone.
・カラム；Intertcap Pure Wax (0.25 i.d. × 0.25 μm × 60 m)
・注入量；1.0 μl
・昇温；50℃で2分保持、2.5℃昇温／分→240℃まで
・気化室温；250℃
・キャリーガス流量；ヘリウム 1.2 ml/分
・スプリット比；1/80
ＧＣＭＳ分析の結果から付属のライブラリー検索（wiley7nおよびNIST08）で化合物をＡＲ－ターメロン、α－ターメロン及びβターメロンと推定した。
ヘキサン抽出物はＡＲ－ターメロンを９．９％、α－ターメロンを３０．４％、及びβ－ターメロンを１１．５％含有していた。 The hexane extract was analyzed using GCMS under the following conditions.
・ Column; Intertcap Pure Wax (0.25 id × 0.25 μm × 60 m)
Injection volume: 1.0 μl
-Temperature rising; holding at 50 ° C for 2 minutes, 2.5 ° C heating / minute to 240 ° C-Vaporization room temperature: 250 ° C
・ Carry gas flow rate: Helium 1.2 ml / min ・ Split ratio: 1/80
From the results of GCMS analysis, the compound was estimated to be AR-turmerone, α-turmerone and β-turmerone by attached library search (wiley7n and NIST08).
The hexane extract contained 9.9% AR-turmerone, 30.4% α-turmerone, and 11.5% β-turmerone.
ヘキサン抽出物を以下のHPLC条件で分画した。
・カラム；L-column ODS （20 id × 250 mm）
・移動相；water:acetonitrile（20：80, v/v）
・流速：18.9 ml/min
・カラム温度；室温
・検出；UV 243 nm
・注入量；10 μl
ヘキサン抽出物の有効成分としてRT７．５、１０．５及び１１．０の画分よりそれぞれ無色のオイル状物質を得た。従来、α－ターメロン及びβ－ターメロンは分離することが難しい物質として知られていたが、上記のような簡便な方法で抽出できたのは特筆すべきことである。 The hexane extract was fractionated under the following HPLC conditions.
・ Column; L-column ODS (20 id × 250 mm)
・ Mobile phase: water: acetonitrile (20: 80, v / v)
・ Flow rate: 18.9 ml / min
・ Column temperature; Room temperature ・ Detection; UV 243 nm
· Injection volume: 10 μl
As active ingredients of the hexane extract, colorless oily substances were obtained from the fractions of RT 7.5, 10.5 and 11.0 respectively. Heretofore, α-turmerone and β-turmerone have been known as substances which are difficult to separate, but it should be noted that they could be extracted by such a simple method as described above.
・カラム；L-column ODS （20 id × 250 mm）
・移動相；water:acetonitrile（20：80, v/v）
・流速：18.9 ml/min
・カラム温度；室温
・検出；UV 243 nm
・注入量；10 μl
ヘキサン抽出物の有効成分としてRT７．５、１０．５及び１１．０の画分よりそれぞれ無色のオイル状物質を得た。従来、α－ターメロン及びβ－ターメロンは分離することが難しい物質として知られていたが、上記のような簡便な方法で抽出できたのは特筆すべきことである。 The hexane extract was fractionated under the following HPLC conditions.
・ Column; L-column ODS (20 id × 250 mm)
・ Mobile phase: water: acetonitrile (20: 80, v / v)
・ Flow rate: 18.9 ml / min
・ Column temperature; Room temperature ・ Detection; UV 243 nm
· Injection volume: 10 μl
As active ingredients of the hexane extract, colorless oily substances were obtained from the fractions of RT 7.5, 10.5 and 11.0 respectively. Heretofore, α-turmerone and β-turmerone have been known as substances which are difficult to separate, but it should be noted that they could be extracted by such a simple method as described above.
上記無色のオイル状物質について、上記段落［００５０］記載のＧＣＭＳ条件で分析し、その結果から付属のライブラリー検索（wiley7nおよびNIST08）およびＮＭＲ解析を用いて化合物がそれぞれＡＲ－ターメロン、α－ターメロン及びβ－ターメロンであることを確認した（Hideji Itokawa,Fusayoshi Hirayama,Kazuko Funakoshi and Koichi Takeya Chem.Pharm.Bull.33(8)3488-3492(1985)及びShin-ichi Uehara,Ichiro Yasuda,Koichi Takeya,Hideji Itokawa Yakugaku zasshi 112(11),817-823の文献値との比較により確認した）。
The above colorless oily substance is analyzed under the GCMS conditions described in the above paragraph [0050], and the results show that the compounds are AR-turmerone and α-turmerone, respectively, using attached library search (wiley7n and NIST08) and NMR analysis. And β-turmerone (Hideji Itokawa, Fusayoshi Hirayama, Kazuko Funakoshi and Koichi Takeya Chem. Pharm. Bull. 33 (8) 3488-3492 (1985) and Shin-ichi Uehara, Ichiro Yasuda, Koichi Takeya, This is confirmed by comparison with literature values of Hideji Itokawa Yakugaku zasshi 112 (11), 817-823).
上記で同定した化合物（ＡＲ－ターメロン、α－ターメロン及びβ－ターメロン）の標準品の酵素阻害活性を、上記の（β‐セクレターゼ阻害率の測定）と同様の方法で検討し、各標準品についてＩＣ５０値を算出した結果、以下の通りであった。
ＡＲ－ターメロン：９２μＭ
α－ターメロン：３９μＭ
β－ターメロン：６２μＭ
上記ＩＣ５０値により、ＡＲ－ターメロン、α－ターメロン及びβ－ターメロンが、β－セクレターゼ阻害活性を有することがわかった。
このＩＣ５０値は、本発明者らが以前に見出し、特願２０１５－１４６２０８において記載したセサモリン、ピパタリン及びエチル４－メトキシシンナメートのＩＣ５０値（セサモリン：０．１４ｍＭ、エチル４－メトキシシンナメート：０．６７６ｍＭ、ピパタリン：０．３０４ｍＭ）と比較しても顕著に低いものであり、ＡＲ－ターメロン、α－ターメロン及びβ－ターメロンは非常に高いβ‐セクレターゼ阻害活性を有することが明らかになった。
また、本発明の抽出方法は特願２０１５－１４０２０８記載の抽出方法と比較して簡便なものであるため、生産コストを１／１０以下に抑えることができる。 The enzyme inhibitory activity of the standard products of the above-identified compounds (AR-turmerone, α-turmerone and β-turmerone) was examined in the same manner as above (measurement of β-secretase inhibition rate), and each standard product was examined. As a result of calculating IC50 value, it was as follows.
AR-turmeron: 92 μM
α-turmelon: 39 μM
β-turmerone: 62 μM
The above-mentioned IC50 value showed that AR-turmerone, α-turmerone and β-turmerone have β-secretase inhibitory activity.
This IC50 value is the IC50 value of sesamoline, pipetalin and ethyl 4-methoxycinnamate which the inventors previously found and described in Japanese Patent Application No. 2015-146208 (sesamoline: 0.14 mM, ethyl 4-methoxycinnamate: 0 AR-turmerone, α-turmerone and β-turmerone were found to have very high β-secretase inhibitory activity, which is significantly lower compared to 676 mM and pipetalin: 0.304 mM).
Further, since the extraction method of the present invention is simple as compared with the extraction method described in Japanese Patent Application No. 2015-140208, the production cost can be suppressed to 1/10 or less.
ＡＲ－ターメロン：９２μＭ
α－ターメロン：３９μＭ
β－ターメロン：６２μＭ
上記ＩＣ５０値により、ＡＲ－ターメロン、α－ターメロン及びβ－ターメロンが、β－セクレターゼ阻害活性を有することがわかった。
このＩＣ５０値は、本発明者らが以前に見出し、特願２０１５－１４６２０８において記載したセサモリン、ピパタリン及びエチル４－メトキシシンナメートのＩＣ５０値（セサモリン：０．１４ｍＭ、エチル４－メトキシシンナメート：０．６７６ｍＭ、ピパタリン：０．３０４ｍＭ）と比較しても顕著に低いものであり、ＡＲ－ターメロン、α－ターメロン及びβ－ターメロンは非常に高いβ‐セクレターゼ阻害活性を有することが明らかになった。
また、本発明の抽出方法は特願２０１５－１４０２０８記載の抽出方法と比較して簡便なものであるため、生産コストを１／１０以下に抑えることができる。 The enzyme inhibitory activity of the standard products of the above-identified compounds (AR-turmerone, α-turmerone and β-turmerone) was examined in the same manner as above (measurement of β-secretase inhibition rate), and each standard product was examined. As a result of calculating IC50 value, it was as follows.
AR-turmeron: 92 μM
α-turmelon: 39 μM
β-turmerone: 62 μM
The above-mentioned IC50 value showed that AR-turmerone, α-turmerone and β-turmerone have β-secretase inhibitory activity.
This IC50 value is the IC50 value of sesamoline, pipetalin and ethyl 4-methoxycinnamate which the inventors previously found and described in Japanese Patent Application No. 2015-146208 (sesamoline: 0.14 mM, ethyl 4-methoxycinnamate: 0 AR-turmerone, α-turmerone and β-turmerone were found to have very high β-secretase inhibitory activity, which is significantly lower compared to 676 mM and pipetalin: 0.304 mM).
Further, since the extraction method of the present invention is simple as compared with the extraction method described in Japanese Patent Application No. 2015-140208, the production cost can be suppressed to 1/10 or less.
本発明は、アルツハイマー型痴呆症を予防するための医薬製剤に好適に使用することができ、清涼飲料水、乳製品、菓子類又はサプリメント等の各種飲食品の添加剤に好適に使用することができる。
INDUSTRIAL APPLICABILITY The present invention can be suitably used for a pharmaceutical preparation for preventing Alzheimer's disease, and can be suitably used for additives of various foods and drinks such as soft drinks, dairy products, confectioneries or supplements. it can.
Claims (4)
- ＡＲ－ターメロン、α－ターメロン及びβ－ターメロンからなる群から選択される１以上の化合物を有効成分として含有するβ‐セクレターゼ阻害剤。 AR-turmerone, a β-secretase inhibitor comprising one or more compounds selected from the group consisting of α-turmerone and β-turmerone as active ingredients.
- ＡＲ－ターメロン、α－ターメロン及びβ－ターメロンが、ターメリックの根茎より抽出されてなることを特徴とする請求項１記載のβ‐セクレターゼ阻害剤。 The beta-secretase inhibitor according to claim 1, wherein the AR-turmerone, the alpha-turmerone and the beta-turmerone are extracted from the rhizome of turmeric.
- 請求項１又は２に記載のβ‐セクレターゼ阻害剤を含む医薬製剤。 A pharmaceutical preparation comprising the β-secretase inhibitor according to claim 1 or 2.
- 請求項１又は２に記載のβ－セクレターゼ阻害剤を含む飲食品。 Food-drinks containing the (beta) -secretase inhibitor of Claim 1 or 2.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2017551474A JP6998212B2 (en) | 2015-11-19 | 2015-11-19 | A β-secretase inhibitor containing turmeron as an active ingredient, and foods and drinks containing the inhibitor. |
| PCT/JP2015/082611 WO2017085844A1 (en) | 2015-11-19 | 2015-11-19 | Β-secretase inhibitor containing turmerone as active ingredient, and pharmaceutical preparation and food/drink containing said inhibitor |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| PCT/JP2015/082611 WO2017085844A1 (en) | 2015-11-19 | 2015-11-19 | Β-secretase inhibitor containing turmerone as active ingredient, and pharmaceutical preparation and food/drink containing said inhibitor |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| WO2017085844A1 true WO2017085844A1 (en) | 2017-05-26 |
Family
ID=58719235
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| PCT/JP2015/082611 WO2017085844A1 (en) | 2015-11-19 | 2015-11-19 | Β-secretase inhibitor containing turmerone as active ingredient, and pharmaceutical preparation and food/drink containing said inhibitor |
Country Status (2)
| Country | Link |
|---|---|
| JP (1) | JP6998212B2 (en) |
| WO (1) | WO2017085844A1 (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2007109210A2 (en) * | 2006-03-17 | 2007-09-27 | Herbalscience Singapore Pte. Ltd. | Extracts and methods comprising curcuma species |
| JP2013189385A (en) * | 2012-03-12 | 2013-09-26 | Kinki Univ | β-SECRETASE INHIBITOR AND FOOD AND DRINK CONTAINING β-SECRETASE INHIBITOR |
| US8859020B2 (en) * | 2005-05-30 | 2014-10-14 | Benny Antony | Treatment of alzheimer's with a curcuminoid mixture and essential oil of turmeric having 45% Ar-turmerone |
-
2015
- 2015-11-19 WO PCT/JP2015/082611 patent/WO2017085844A1/en active Application Filing
- 2015-11-19 JP JP2017551474A patent/JP6998212B2/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US8859020B2 (en) * | 2005-05-30 | 2014-10-14 | Benny Antony | Treatment of alzheimer's with a curcuminoid mixture and essential oil of turmeric having 45% Ar-turmerone |
| WO2007109210A2 (en) * | 2006-03-17 | 2007-09-27 | Herbalscience Singapore Pte. Ltd. | Extracts and methods comprising curcuma species |
| JP2013189385A (en) * | 2012-03-12 | 2013-09-26 | Kinki Univ | β-SECRETASE INHIBITOR AND FOOD AND DRINK CONTAINING β-SECRETASE INHIBITOR |
Non-Patent Citations (2)
| Title |
|---|
| DOUGLAS R. SHYTLE ET AL.: "Optimized turmeric extracts have potent anti-amyloidogenic effects", CURRENT ALZHEIMER RESEARCH, vol. 6, 2009, pages 564 - 571, ISSN: 1567-2050 * |
| SUN YOUNG PARK ET AL.: "Anti-inflammatory effects of aromatic-turmerone through blocking of NF-KB, JNK, and p38 MAPK signaling pathways in amyloid beta-stimulated microglia", INTERNATIONAL IMMUNOPHARMACOLOGY, vol. 14, no. 1, 20 June 2012 (2012-06-20), pages 13 - 20, XP028425798, ISSN: 1567-5769 * |
Also Published As
| Publication number | Publication date |
|---|---|
| JPWO2017085844A1 (en) | 2018-09-13 |
| JP6998212B2 (en) | 2022-01-18 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| JP6004419B2 (en) | Foods and beverages containing β-secretase inhibitors and β-secretase inhibitors | |
| JP6447677B2 (en) | Composition for preventing and / or treating cognitive impairment | |
| US20130178436A1 (en) | Composition for preventing, improving, or treating renal disease including maillard browning reaction products of panax species plant extract | |
| KR101776001B1 (en) | Composition for treating, preventing or improving born loss containing panduratin or fingerroot(boesenbergia pandurata) extract | |
| JP5275315B2 (en) | Novel compound and β-secretase inhibitor | |
| KR20210089104A (en) | Composition for preventing and treating inflammatory Lung diseases caused by ultrafine dust containing Porphyra tenera extract as an active ingredient | |
| KR102191279B1 (en) | Pharmaceutical composition for prevention or treatment inflammatory diseases comprising comprising root extract Hovenia dulcis, or fractions thereof, or compounds isolated from therefrom | |
| WO2017085844A1 (en) | Β-secretase inhibitor containing turmerone as active ingredient, and pharmaceutical preparation and food/drink containing said inhibitor | |
| JP2020125327A (en) | β-SECRETASE INHIBITOR CONTAINING TURMERONE AS ACTIVE INGREDIENT, AND PHARMACEUTICAL PREPARATION AND FOOD/DRINK CONTAINING SAID INHIBITOR | |
| JP6383334B2 (en) | β-secretase inhibitor, and pharmaceutical preparation and food and drink containing the inhibitor | |
| JP2012131760A (en) | Fatty acid absorption inhibitor | |
| KR20140128710A (en) | Pharmaceutical composition for prevention or treatment of inflammatory or allergic diseases comprising the extract Cinnamomum cambodianum or compound marliolide isolated therefrom as an active ingredient | |
| TW201943427A (en) | Composition for inhibiting proliferation of cancer cells, anticancer composition, composition for preventing cancerization of normal cells, composition for preventing cancer onset, and composition for inducing cancer cell death | |
| KR101454336B1 (en) | Compositions for preventing and treating arthritis | |
| JP2013193997A (en) | Angiotensin ii 1 type receptor antagonist and hypotensive agent | |
| JP4828135B2 (en) | Inhibitor of cholesterol uptake into lipid micelles | |
| JP6142936B2 (en) | β-secretase inhibitor and food and drink for β-secretase inhibition | |
| JP6142935B2 (en) | β-secretase inhibitor and food and drink for β-secretase inhibition | |
| KR101964329B1 (en) | Composition for preventing or treating of cancer comprising saringosterol acetate from brown algae or Hizikia fusiformis | |
| KR20200131209A (en) | Pharmaceutical composition for prevention or treatment inflammatory diseases comprising comprising root extract Hovenia dulcis, or fractions thereof, or compounds isolated from therefrom | |
| KR20230039887A (en) | Composition for preventing or treating inflammatory raspiratory diseases caused by ultrafine dust containing mugwort and lizard’s tail extract as an active ingredient | |
| KR101651106B1 (en) | ISOLATED SINGLE COMPOUND FROM Mori Cortex Radicis ITS APPLICATION IN TREATING AND PREVENTING OBESITY | |
| KR20210087771A (en) | Composition for preventing or treating dementia comprising Houttuynia cordata or isolated compounds of Houttuynia cordata | |
| KR20150103847A (en) | Composition for prevention or treatment of nonalcoholic fatty liver disease containing Puerariae Flos extract | |
| KR20140107992A (en) | Pharmaceutical composition for prevention or treatment of Parkinson's disease comprising Eucommiae ulmoides extract or fraction thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| 121 | Ep: the epo has been informed by wipo that ep was designated in this application |
Ref document number: 15908784 Country of ref document: EP Kind code of ref document: A1 |
|
| DPE1 | Request for preliminary examination filed after expiration of 19th month from priority date (pct application filed from 20040101) | ||
| ENP | Entry into the national phase |
Ref document number: 2017551474 Country of ref document: JP Kind code of ref document: A |
|
| NENP | Non-entry into the national phase |
Ref country code: DE |
|
| 122 | Ep: pct application non-entry in european phase |
Ref document number: 15908784 Country of ref document: EP Kind code of ref document: A1 |