JP2009067749A - Matrix metalloprotease-1(mmp-1) inhibitor, matrix metalloprotease-2(mmp-2) inhibitor, estrogen-like action agent, profilaggrin production promoter, filaggrin production promoter, antiobese agent and cyclic amp phosphodiesterase activity inhibitor - Google Patents
Matrix metalloprotease-1(mmp-1) inhibitor, matrix metalloprotease-2(mmp-2) inhibitor, estrogen-like action agent, profilaggrin production promoter, filaggrin production promoter, antiobese agent and cyclic amp phosphodiesterase activity inhibitor Download PDFInfo
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- JP2009067749A JP2009067749A JP2007239859A JP2007239859A JP2009067749A JP 2009067749 A JP2009067749 A JP 2009067749A JP 2007239859 A JP2007239859 A JP 2007239859A JP 2007239859 A JP2007239859 A JP 2007239859A JP 2009067749 A JP2009067749 A JP 2009067749A
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- inhibitor
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- acteoside
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Abstract
Description
本発明は、マトリックスメタロプロテアーゼ−１（ＭＭＰ−１）阻害剤、マトリックスメタロプロテアーゼ−２（ＭＭＰ−２）阻害剤、エストロゲン様作用剤、プロフィラグリン産生促進剤、フィラグリン産生促進剤、抗肥満剤及びサイクリックＡＭＰホスホジエステラーゼ活性阻害剤に関するものである。 The present invention relates to a matrix metalloproteinase-1 (MMP-1) inhibitor, a matrix metalloproteinase-2 (MMP-2) inhibitor, an estrogen-like agent, a profilagrin production promoter, a filaggrin production promoter, an antiobesity agent, and The present invention relates to a cyclic AMP phosphodiesterase activity inhibitor.
近年、皮膚の老化に伴う変化を誘導する因子として、マトリックスメタロプロテアーゼ（ＭＭＰｓ；Matrix metalloproteinases）の関与が指摘されている。このＭＭＰｓの中でも、マトリックスメタロプロテアーゼ−１（ＭＭＰ−１）は、皮膚の真皮細胞外マトリックスの主要構成成分であるコラーゲンを分解する酵素として知られているが、その発現は紫外線の照射により大きく増加し、コラーゲンの減少・変性の一因となり、皮膚のシワの形成、弾力性の低下等の大きな要因となると考えられている。したがって、ＭＭＰ−１の活性を阻害することは、皮膚の老化症状を予防・改善する上で重要である。このようなＭＭＰ−１阻害作用を有するものとしては、例えば、マツ科マツ属二葉松類の樹皮の抽出物等が知られている（特許文献１参照）。 In recent years, the involvement of matrix metalloproteinases (MMPs) has been pointed out as factors that induce changes associated with skin aging. Among these MMPs, matrix metalloproteinase-1 (MMP-1) is known as an enzyme that degrades collagen, which is a major component of the dermal extracellular matrix of skin, but its expression is greatly increased by irradiation with ultraviolet rays. However, it is thought to contribute to the decrease / denaturation of collagen, and to be a major factor such as the formation of wrinkles on the skin and the decrease in elasticity. Therefore, inhibiting the activity of MMP-1 is important in preventing and improving skin aging symptoms. For example, an extract of the bark of the Pinus pine genus Futaba is known as one having such an MMP-1 inhibitory effect (see Patent Document 1).
また、ＭＭＰｓの中でも、ゼラチナーゼ群に属する酵素であるマトリックスメタロプロテアーゼ−２（ＭＭＰ−２）は、基底膜の主要構成成分であるＩＶ型コラーゲンやラミニン５を分解する酵素として知られているが、その発現及び活性は紫外線の照射により大きく増加し、紫外線による基底膜成分の減少、基底膜の構造変化の原因となり、皮膚におけるシワやたるみの形成等の大きな要因となることが明らかとなっている（非特許文献１参照）。 Among MMPs, matrix metalloproteinase-2 (MMP-2), an enzyme belonging to the gelatinase group, is known as an enzyme that degrades type IV collagen and laminin 5, which are the main components of the basement membrane. Its expression and activity are greatly increased by ultraviolet irradiation, and it has been clarified that it causes a decrease in basement membrane components and structural changes of the basement membrane due to ultraviolet rays, and it is a major factor in the formation of wrinkles and sagging in the skin. (Refer nonpatent literature 1).
さらに、マトリックスメタロプロテアーゼ−２（ＭＭＰ−２）は、血管内皮細胞下に存在する基底膜を構成するＩＶ型コラーゲン等を分解し、分解された血管内皮細胞は間質へ遊走していき、間質中で増殖し、管腔を形成し、新生血管を構築していく。そして、この新生された血管が腫瘍細胞に到達して、栄養源と酸素とを腫瘍細胞に供給し、腫瘍が大きくなっていくことが知られている（非特許文献２参照）。 Furthermore, matrix metalloproteinase-2 (MMP-2) degrades type IV collagen and the like that make up the basement membrane present under vascular endothelial cells, and the degraded vascular endothelial cells migrate to the stroma. Proliferates in the fluid, forms lumens, and builds new blood vessels. Then, it is known that the newly formed blood vessels reach the tumor cells, supply nutrient sources and oxygen to the tumor cells, and the tumor grows (see Non-Patent Document 2).
したがって、ＭＭＰ−２の活性を阻害することにより、基底膜成分の減少、基底膜の構造変化を抑制し、皮膚機能を改善することができるとともに、血管新生を抑制し、腫瘍細胞の増殖を抑制することができると考えられる。このようなＭＭＰ−２阻害作用を有するものとしては、例えば、黒霊芝抽出物（特許文献２参照）、クリ渋皮発酵物からの抽出物（特許文献３参照）等が知られている。 Therefore, by inhibiting the activity of MMP-2, it is possible to suppress the decrease in basement membrane components and structural changes of the basement membrane, improve the skin function, suppress angiogenesis, and suppress the growth of tumor cells. I think it can be done. As what has such an MMP-2 inhibitory effect, the extract from a black reishi extract (refer patent document 2), an extract from chestnut astringent skin fermented product (refer patent document 3), etc. are known, for example.
加齢に伴う皮膚老化の一因は、女性ホルモンの一種であるエストロゲンの分泌が減退することにある。すなわち、エストロゲンは成人女性の健康維持に深く関わっており、その分泌不足は種々の内科的疾患を招くほか、肌の過敏症、弾力性の低下、潤いの減少等、好ましくない肌の変化の原因となったり、閉経後の女性等におけるエストロゲンの欠乏は、冠動脈性心臓疾患や骨粗鬆症の原因となったりすることが知られている。 One cause of skin aging with aging is a decrease in the secretion of estrogen, a female hormone. In other words, estrogen is deeply involved in maintaining the health of adult women, and its lack of secretion leads to various medical illnesses, as well as causes of unfavorable skin changes such as skin hypersensitivity, reduced elasticity, reduced moisture, etc. It is known that estrogen deficiency in postmenopausal women and the like causes coronary heart disease and osteoporosis.
そこで、エストロゲンの分泌が衰える更年期以降の女性に対して、エストロゲンと同様の作用を有する物質を配合した薬剤を、経皮的又は経口的に投与することが行われている。このようなエストロゲン様作用を有するものとしては、例えば、五斂子の葉部からの抽出物（特許文献４参照）等が知られている。 Therefore, a drug containing a substance having the same action as estrogen is transdermally or orally administered to women after menopause, whose estrogen secretion declines. As what has such an estrogen-like effect | action, the extract (refer patent document 4) etc. from the leaf part of a quince is known, for example.
天然保湿因子（Natural Moisturizing Factors；ＮＭＦ）の主成分であるアミノ酸は、ケラトヒアリン顆粒に由来するフィラグリンが角質層内で分解されて産生される。このフィラグリンは、角質層直下の顆粒層に存在する表皮ケラチノサイトでプロフィラグリンとして発現する。その後、直ちにリン酸化し、ケラトヒアリン顆粒に蓄積され、脱リン酸，加水分解を経てフィラグリンへと分解され、角質層に移行して、ケラチンフィラメントの凝集効率を高め、角質細胞の内部構築に関与することが知られている（非特許文献３参照）。 An amino acid which is a main component of natural moisturizing factors (NMF) is produced by degrading filaggrin derived from keratohyalin granules in the stratum corneum. This filaggrin is expressed as profilagrin in epidermal keratinocytes existing in the granule layer immediately below the stratum corneum. It is then phosphorylated immediately, accumulated in keratohyalin granules, decomposed to filaggrin through dephosphorylation and hydrolysis, transferred to the stratum corneum, increased the efficiency of keratin filament aggregation, and is involved in the internal construction of keratinocytes It is known (see Non-Patent Document 3).
近年、このフィラグリンが、皮膚の水分保持に非常に重要かつ必要不可欠であること、及び乾燥等の条件によってフィラグリンの合成力が低下し、角質層におけるアミノ酸量が低下することが知られている（非特許文献４参照）。 In recent years, it has been known that this filaggrin is very important and indispensable for moisture retention of the skin, and that the synthetic power of filaggrin is reduced by conditions such as drying, and the amount of amino acids in the stratum corneum is reduced ( Non-patent document 4).
したがって、表皮ケラチノサイトにおいて、プロフィラグリンｍＲＮＡの発現促進を通じて、フィラグリンの産生を促進し、それにより角質層内のアミノ酸量を増大させることで、角質層の水分環境を本質的に改善することができると期待されている。 Therefore, in epidermal keratinocytes, it is possible to essentially improve the water environment of the stratum corneum by promoting filaggrin production through promoting the expression of profilagrin mRNA, thereby increasing the amount of amino acids in the stratum corneum. Expected.
このようなプロフィラグリン産生促進作用及びフィラグリン産生促進作用を有するものとしては、例えば、カンゾウ抽出物（特許文献５参照）、天然植物中に含まれるフラバノン配糖体として知られるリクイリチン（特許文献６参照）等が知られており、プロフィラグリン及びフィラグリン蛋白産生促進作用を有するものとしては、例えば、Citrus属に属する植物エキス又は酵母エキス（特許文献７参照）等が知られている。 As what has such a profilagulin production promotion effect and a filaggrin production promotion effect, for example, a liquorice extract (refer patent document 5), the liquiritin known as flavanone glycoside contained in a natural plant (refer patent document 6) For example, a plant extract or yeast extract belonging to the genus Citrus (see Patent Document 7) or the like is known as one having profilaggrin and filaggrin protein production promoting action.
近年、飽食や運動不足等の生活習慣が原因となって体脂肪が増加し、肥満が増えている。このような肥満の増加は、人間ばかりでなく、ペットや家畜においても見られる。肥満は、高脂血症や動脈硬化等の成人病の原因になるため、美容の面で問題となるばかりでなく、健康の面でも大きな問題となる。 In recent years, body fat has increased due to lifestyle habits such as satiety and lack of exercise, and obesity has increased. Such an increase in obesity is seen not only in humans but also in pets and livestock. Obesity is a cause of adult diseases such as hyperlipidemia and arteriosclerosis, which is not only a cosmetic problem but also a major problem in health.
生体内の脂肪を分解するためには、サイクリックＡＭＰの役割が重要となる。サイクリックＡＭＰは生体内に存在するリパーゼを活性化し、活性化されたリパーゼによって脂肪が脂肪酸とグリセロールとに分解される。しかし、サイクリックＡＭＰホスホジエステラーゼが活性化されるとサイクリックＡＭＰの分解が誘発され、リパーゼの活性化が阻害される。そのため、サイクリックＡＭＰホスホジエステラーゼの活性を阻害することにより細胞内におけるサイクリックＡＭＰが増量し、脂肪の分解を促進することができるものと考えられる。このようなサイクリックＡＭＰホスホジエステラーゼ活性阻害作用を有するものとして、ピーナッツ渋皮抽出物（特許文献８参照）等が知られている。
本発明は第一に、安全性の高い天然物の中からマトリックスメタロプロテアーゼ−１活性阻害作用を有するものを見出し、それを有効成分とするマトリックスメタロプロテアーゼ−１阻害剤を提供することを目的とする。 It is an object of the present invention to provide a matrix metalloproteinase-1 inhibitor having an active ingredient as a matrix metalloprotease-1 inhibitory substance, which is a highly safe natural product. To do.
本発明は第二に、安全性の高い天然物の中からマトリックスメタロプロテアーゼ−２活性阻害作用を有するものを見出し、それを有効成分とするマトリックスメタロプロテアーゼ−２阻害剤を提供することを目的とする。 A second object of the present invention is to provide a matrix metalloproteinase-2 inhibitor having an active ingredient as a matrix metalloprotease-2 inhibitory activity from among highly safe natural products. To do.
本発明は第三に、安全性の高い天然物の中からエストロゲン様作用を有するものを見出し、それを有効成分とするエストロゲン様作用剤を提供することを目的とする。 A third object of the present invention is to find an estrogen-like agent having an estrogen-like action from highly safe natural products, and to provide an estrogen-like agent containing the same as an active ingredient.
本発明は第四に、安全性の高い天然物の中からプロフィラグリン産生促進作用を有するものを見出し、それを有効成分とするプロフィラグリン産生促進剤を提供することを目的とする。 A fourth object of the present invention is to find a profilagline production promoting agent from among highly safe natural products, and to provide a profilagrin production promoter containing the same as an active ingredient.
本発明は第五に、安全性の高い天然物の中からフィラグリン産生促進作用を有するものを見出し、それを有効成分とするフィラグリン産生促進剤を提供することを目的とする。 The fifth object of the present invention is to find a filaggrin production promoting action from highly safe natural products, and to provide a filaggrin production promoter containing the same as an active ingredient.
本発明は第六に、安全性の高い天然物の中からサイクリックＡＭＰホスホジエステラーゼ阻害作用を有するものを見出し、それを有効成分とする抗肥満剤及びサイクリックＡＭＰホスホジエステラーゼ阻害剤を提供することを目的とする。 A sixth object of the present invention is to find a highly safe natural product having a cyclic AMP phosphodiesterase inhibitory activity, and to provide an anti-obesity agent and a cyclic AMP phosphodiesterase inhibitor containing the same as an active ingredient. And
上記課題を解決するため、本発明のマトリックスメタロプロテアーゼ−１阻害剤、マトリックスメタロプロテアーゼ−２阻害剤、エストロゲン様作用剤、プロフィラグリン産生促進剤、フィラグリン産生促進剤、抗肥満剤及びサイクリックＡＭＰホスホジエステラーゼ阻害剤は、キンモクセイからの抽出物及び／又はアクテオシド（Acteoside）を有効成分として含有することを特徴とする。 In order to solve the above problems, the matrix metalloproteinase-1 inhibitor, matrix metalloproteinase-2 inhibitor, estrogen-like agent, profilagrin production promoter, filaggrin production promoter, anti-obesity agent and cyclic AMP phosphodiesterase of the present invention The inhibitor is characterized by containing, as an active ingredient, an extract from Buttercup and / or Acteoside.
本発明によれば、天然物であるキンモクセイからの抽出物及び／又はアクテオシド（Acteoside）を有効成分として含有し、安全性に優れたマトリックスメタロプロテアーゼ−１阻害剤、マトリックスメタロプロテアーゼ−２阻害剤、エストロゲン様作用剤、プロフィラグリン産生促進剤、フィラグリン産生促進剤、抗肥満剤及びサイクリックＡＭＰホスホジエステラーゼ阻害剤を提供することができる。 According to the present invention, a matrix metalloproteinase-1 inhibitor, a matrix metalloprotease-2 inhibitor, which contains an extract from a natural venom and / or Acteoside as an active ingredient and is excellent in safety, An estrogen-like agent, a profilagrin production promoter, a filaggrin production promoter, an anti-obesity agent, and a cyclic AMP phosphodiesterase inhibitor can be provided.
以下、本発明について説明する。
本発明のマトリックスメタロプロテアーゼ−１阻害剤、マトリックスメタロプロテアーゼ−２阻害剤、エストロゲン様作用剤、プロフィラグリン産生促進剤、フィラグリン産生促進剤、抗肥満剤又はサイクリックＡＭＰホスホジエステラーゼ阻害剤は、キンモクセイからの抽出物及び／又はアクテオシド（Acteoside）を有効成分として含有する。
The present invention will be described below.
Matrix metalloprotease-1 inhibitor, matrix metalloproteinase-2 inhibitor, estrogen-like agent, profilagrin production promoter, filaggrin production promoter, anti-obesity agent or cyclic AMP phosphodiesterase inhibitor of the present invention It contains an extract and / or Acteoside as an active ingredient.
ここで、本発明において「キンモクセイからの抽出物」には、キンモクセイを抽出原料として得られる抽出液、当該抽出液の希釈液若しくは濃縮液、当該抽出液を乾燥して得られる乾燥物、又はこれらの粗精製物若しくは精製物のいずれもが含まれる。 Here, in the present invention, the “extract from Buttercup” refers to an extract obtained by using a Snapper as an extraction raw material, a diluted or concentrated solution of the extract, a dried product obtained by drying the extract, or these Both of these crudely purified products and purified products are included.
アクテオシド（Acteoside）は、下記式（Ｉ）で表される化学構造を有するポリフェノールである。 Acteoside is a polyphenol having a chemical structure represented by the following formula (I).
アクテオシドは、アクテオシドを含有する植物抽出物から単離・精製することにより製造することができる。このようなアクテオシドを含有する植物抽出物は、植物の抽出に一般に用いられている方法によって得ることができる。アクテオシドを含有する植物としては、例えば、キンモクセイ（学名：Osmanthus fragrans var. aurantiacus）等が挙げられる。 Acteoside can be produced by isolation and purification from a plant extract containing acteoside. Such a plant extract containing acteoside can be obtained by a method generally used for plant extraction. Examples of the plant containing acteoside include, for example, ummingei (scientific name: Osmanthus fragrans var. Aurantiacus).
キンモクセイ（Osmanthus fragrans var. aurantiacus）は、モクセイ科モクセイ属に属する常緑小高木であり、別名、桂花とも呼ばれ、中国南部が原産地であり、このような地域から容易に入手することができる。抽出原料として使用し得るキンモクセイの構成部位としては、例えば、葉部、枝部、樹皮部、幹部、茎部、果実部、花部等の地上部、根部又はこれらの部位の混合物等が挙げられるが、好ましくは花部である。 Buttercup (Osmanthus fragrans var. Aurantiacus) is an evergreen small tree belonging to the genus Moxae, also known as Katsura, which is native to southern China and can be easily obtained from such areas. Examples of the constituent parts of the cypress that can be used as the raw material for extraction include the aerial parts such as leaves, branches, bark parts, trunk parts, stem parts, fruit parts, flower parts, root parts, or a mixture of these parts. However, it is preferably a flower part.
ここで、「花」とは、一般に、種子植物の有性生殖にかかわる器官の総体をいい、葉の変形である花葉と茎の変形である花軸とから構成され、花葉には、萼、花弁、雄しべ、心皮等の器官が含まれる。本発明において抽出原料として使用する「花部」には、種子植物の有性生殖にかかわる器官の総体の他、その一部、例えば、花葉、花被（萼と花冠）、花冠、花弁等も含まれる。 Here, the "flower" generally refers to the whole organ involved in sexual reproduction of a seed plant, and is composed of a flower leaf that is a leaf deformation and a flower axis that is a stem deformation. This includes organs such as pupae, petals, stamens, and heart skin. The “floral part” used as an extraction raw material in the present invention includes all of the organs involved in sexual reproduction of seed plants, as well as parts thereof, such as flower leaves, flower coats (buds and corolla), corolla, petals, etc. Is also included.
アクテオシドを含有する植物抽出物は、抽出原料を乾燥した後、そのまま又は粗砕機を用いて粉砕し、抽出溶媒による抽出に供することにより得ることができる。乾燥は天日で行ってもよいし、通常使用される乾燥機を用いて行ってもよい。また、ヘキサン等の非極性溶媒によって脱脂等の前処理を施してから抽出原料として使用してもよい。脱脂等の前処理を行うことにより、キンモクセイの極性溶媒による抽出処理を効率よく行うことができる。 A plant extract containing acteoside can be obtained by drying the raw material for extraction and then pulverizing the raw material as it is or using a crusher and subjecting it to extraction with an extraction solvent. Drying may be performed in the sun or using a commonly used dryer. Moreover, after performing pretreatment, such as degreasing, with a nonpolar solvent such as hexane, it may be used as an extraction raw material. By performing pretreatment such as degreasing, extraction treatment with a polar solvent of cinnabar can be performed efficiently.
抽出溶媒としては、極性溶媒を用いるのが好ましく、例えば、水、親水性有機溶媒等が挙げられ、これらを単独で又は２種以上を組み合わせて、室温又は溶媒の沸点以下の温度で使用することが好ましい。 As the extraction solvent, it is preferable to use a polar solvent, and examples thereof include water and hydrophilic organic solvents. These may be used alone or in combination of two or more at room temperature or a temperature below the boiling point of the solvent. Is preferred.
抽出溶媒として使用し得る水としては、純水、水道水、井戸水、鉱泉水、鉱水、温泉水、湧水、淡水等のほか、これらに各種処理を施したものが含まれる。水に施す処理としては、例えば、精製、加熱、殺菌、濾過、イオン交換、浸透圧調整、緩衝化等が含まれる。したがって、本発明において抽出溶媒として使用し得る水には、精製水、熱水、イオン交換水、生理食塩水、リン酸緩衝液、リン酸緩衝生理食塩水等も含まれる。 Examples of water that can be used as the extraction solvent include pure water, tap water, well water, mineral spring water, mineral water, hot spring water, spring water, fresh water, and the like, and those subjected to various treatments. Examples of the treatment applied to water include purification, heating, sterilization, filtration, ion exchange, osmotic pressure adjustment, buffering, and the like. Therefore, the water that can be used as the extraction solvent in the present invention includes purified water, hot water, ion-exchanged water, physiological saline, phosphate buffer, phosphate buffered saline, and the like.
抽出溶媒として使用することのできる親水性有機溶媒としては、メタノール、エタノール、プロピルアルコール、イソプロピルアルコール等の炭素数１〜５の低級脂肪族アルコール；アセトン、メチルエチルケトン等の低級脂肪族ケトン；１，３−ブチレングリコール、プロピレングリコール、グリセリン等の炭素数２〜５の多価アルコール等が挙げられる。 Examples of hydrophilic organic solvents that can be used as the extraction solvent include lower aliphatic alcohols having 1 to 5 carbon atoms such as methanol, ethanol, propyl alcohol, and isopropyl alcohol; lower aliphatic ketones such as acetone and methyl ethyl ketone; -C2-C5 polyhydric alcohols, such as butylene glycol, propylene glycol, and glycerol, etc. are mentioned.
２種以上の極性溶媒の混合液を抽出溶媒として使用する場合、その混合比は適宜調整することができる。例えば、水と低級脂肪族アルコールとの混合液を使用する場合には、水１０質量部に対して低級脂肪族アルコール１〜９０質量部を混合することが好ましく、水と低級脂肪族ケトンとの混合液を使用する場合には、水１０質量部に対して低級脂肪族ケトン１〜４０質量部を混合することが好ましく、水と多価アルコールとの混合液を使用する場合には、水１０質量部に対して多価アルコール１０〜９０質量部を混合することが好ましい。 When using the liquid mixture of 2 or more types of polar solvents as an extraction solvent, the mixing ratio can be adjusted suitably. For example, when using a liquid mixture of water and a lower aliphatic alcohol, it is preferable to mix 1 to 90 parts by weight of a lower aliphatic alcohol with respect to 10 parts by weight of water. When using a mixed solution, it is preferable to mix 1 to 40 parts by mass of a lower aliphatic ketone with 10 parts by mass of water, and when using a mixed solution of water and a polyhydric alcohol, water 10 It is preferable to mix 10 to 90 parts by mass of polyhydric alcohol with respect to parts by mass.
抽出処理は、抽出原料に含まれる可溶性成分を抽出溶媒に溶出させ得る限り特に限定はされず、常法に従って行うことができる。例えば、抽出原料の５〜１５倍量（質量比）の抽出溶媒に、抽出原料を浸漬し、常温又は還流加熱下で可溶性成分を抽出させた後、濾過して抽出残渣を除去することにより抽出液を得ることができる。得られた抽出液から溶媒を留去するとペースト状の濃縮物が得られ、この濃縮物をさらに乾燥すると乾燥物が得られる。 The extraction treatment is not particularly limited as long as the soluble component contained in the extraction raw material can be eluted in the extraction solvent, and can be performed according to a conventional method. For example, the extraction raw material is immersed in an extraction solvent 5 to 15 times (mass ratio) of the extraction raw material, the soluble components are extracted at room temperature or under reflux, and then filtered to remove the extraction residue. A liquid can be obtained. When the solvent is distilled off from the obtained extract, a paste-like concentrate is obtained, and when this concentrate is further dried, a dried product is obtained.
得られた抽出液はそのままでもマトリックスメタロプロテアーゼ−１阻害剤、マトリックスメタロプロテアーゼ−２阻害剤、エストロゲン様作用剤、プロフィラグリン産生促進剤、フィラグリン産生促進剤、抗肥満剤又はサイクリックＡＭＰホスホジエステラーゼ阻害剤の有効成分として使用することができるが、濃縮液又は乾燥物としたものの方が使用しやすい。 Even if the obtained extract is used as it is, it is a matrix metalloproteinase-1 inhibitor, matrix metalloproteinase-2 inhibitor, estrogen-like agent, profilagrin production promoter, filaggrin production promoter, anti-obesity agent or cyclic AMP phosphodiesterase inhibitor. However, it is easier to use a concentrated or dried product.
キンモクセイからの抽出物は特有の匂いを有しているため、その生理活性の低下を招かない範囲で脱色、脱臭等を目的とする精製を行うことも可能であるが、飲食品等に配合する場合には大量に使用するものではないから、未精製のままでも実用上支障はない。 Since the extract from the buttercup has a peculiar odor, it can be purified for the purpose of decoloring, deodorizing, etc. within a range that does not cause a decrease in its physiological activity, but it is blended in foods and drinks etc. In some cases, it is not used in large quantities, so there is no practical problem even if it is not purified.
以上のようにして得られた抽出液、当該抽出液の濃縮物又は当該抽出液の乾燥物からアクテオシドを単離・精製する方法は、特に限定されるものではなく、常法により行うことができる。例えば、植物抽出物を、シリカゲルやアルミナ等の多孔質物質、スチレン−ジビニルベンゼン共重合体やポリメタクリレート等の多孔性樹脂等を用いたカラムクロマトグラフィーに付して、水、アルコールの順で溶出させることで、アルコールで溶出される画分としてアクテオシドを得ることができる。カラムクロマトグラフィーにて溶出液として用いられるアルコールは、特に限定されるものではなく、例えば、メタノール、エタノール、プロピルアルコール、イソプロピルアルコール等の炭素数１〜５の低級脂肪族アルコール又はそれらの水溶液等が挙げられる。さらに、カラムクロマトグラフィーにより得られたアルコール画分を、ＯＤＳを用いた逆相シリカゲルクロマトグラフィー、再結晶、液−液向流抽出、イオン交換樹脂を用いたカラムクロマトグラフィー等の任意の有機化合物精製手段を用いて精製してもよい。 The method for isolating and purifying acteoside from the extract obtained as described above, the concentrate of the extract or the dried product of the extract is not particularly limited, and can be performed by a conventional method. . For example, a plant extract is subjected to column chromatography using a porous material such as silica gel or alumina, a porous resin such as styrene-divinylbenzene copolymer or polymethacrylate, and eluted in the order of water and alcohol. Thus, acteoside can be obtained as a fraction eluted with alcohol. The alcohol used as the eluent in the column chromatography is not particularly limited, and examples thereof include lower aliphatic alcohols having 1 to 5 carbon atoms such as methanol, ethanol, propyl alcohol, isopropyl alcohol, and their aqueous solutions. Can be mentioned. Further, the alcohol fraction obtained by column chromatography can be purified by any organic compound such as reverse phase silica gel chromatography using ODS, recrystallization, liquid-liquid countercurrent extraction, column chromatography using ion exchange resin, etc. It may be purified using means.
上記のようにして得られるキンモクセイからの抽出物又はアクテオシドは、マトリックスメタロプロテアーゼ−１阻害作用、マトリックスメタロプロテアーゼ−２阻害作用、エストロゲン様作用、プロフィラグリン産生促進作用、フィラグリン産生促進作用、抗肥満作用及びサイクリックＡＭＰホスホジエステラーゼ阻害作用を有しているため、それぞれの作用を利用してマトリックスメタロプロテアーゼ−１阻害剤、マトリックスメタロプロテアーゼ−２阻害剤、エストロゲン様作用剤、プロフィラグリン産生促進剤、フィラグリン産生促進剤、抗肥満剤又はサイクリックＡＭＰホスホジエステラーゼ阻害剤の有効成分として用いることができる。 The extract or acteoside obtained from the above-mentioned beetle is as follows: matrix metalloproteinase-1 inhibitory action, matrix metalloproteinase-2 inhibitory action, estrogen-like action, profilagrin production promoting action, filaggrin production promoting action, anti-obesity action And a cyclic AMP phosphodiesterase inhibitory action, the matrix metalloprotease-1 inhibitor, matrix metalloprotease-2 inhibitor, estrogen-like agent, profilaggrin production promoter, filaggrin production It can be used as an active ingredient of an accelerator, an anti-obesity agent or a cyclic AMP phosphodiesterase inhibitor.
また、キンモクセイからの抽出物又はアクテオシドは、マトリックスメタロプロテアーゼ−１阻害作用、マトリックスメタロプロテアーゼ−２阻害作用、エストロゲン様作用、プロフィラグリン産生促進作用、フィラグリン産生促進作用及びサイクリックＡＭＰホスホジエステラーゼ阻害作用を有しているため、マトリックスメタロプロテアーゼ−１の発現上昇に起因する疾患の予防・治療剤、マトリックスメタロプロテアーゼ−２の発現上昇に起因する疾患の予防・治療剤、エストロゲンの欠乏に起因する疾患の予防・治療剤、プロフィラグリンの産生量低下に起因する疾患の予防・治療剤、フィラグリンの産生量低下に起因する疾患の予防・治療剤、又はサイクリックＡＭＰの産生量低下に起因する疾患の予防・治療剤の有効成分として使用することができる。 In addition, the extract or acteoside from the venom has a matrix metalloproteinase-1 inhibitory action, a matrix metalloproteinase-2 inhibitory action, an estrogen-like action, a profilagulin production promoting action, a filaggrin production promoting action and a cyclic AMP phosphodiesterase inhibiting action. Therefore, a preventive / therapeutic agent for diseases caused by increased expression of matrix metalloproteinase-1, a preventive / therapeutic agent for diseases caused by increased expression of matrix metalloproteinase-2, and a prevention of diseases caused by deficiency of estrogen・ Therapeutic agent, prevention / treatment agent for diseases caused by decreased production of profilagrin, prevention / treatment agent for diseases caused by reduced production of filaggrin, or prevention of diseases caused by reduced production of cyclic AMP ・As an active ingredient of a therapeutic agent It is possible to use.
マトリックスメタロプロテアーゼ−１阻害剤、マトリックスメタロプロテアーゼ−２阻害剤、エストロゲン様作用剤、プロフィラグリン産生促進剤、フィラグリン産生促進剤、抗肥満剤又はサイクリックＡＭＰホスホジエステラーゼ阻害剤は、有効成分としてキンモクセイからの抽出物のみを含有していてもよいし、アクテオシドのみを含有していてもよいし、キンモクセイからの抽出物とアクテオシドとの混合物を含有していてもよい。キンモクセイからの抽出物とアクテオシドとの混合物を有効成分として含有せしめる場合、その配合割合は、キンモクセイからの抽出物やアクテオシドの有する作用の程度に応じて適宜決定すればよい。 Matrix metalloprotease-1 inhibitor, matrix metalloprotease-2 inhibitor, estrogen-like agent, profilaggrin production promoter, filaggrin production promoter, anti-obesity agent or cyclic AMP phosphodiesterase inhibitor is an active ingredient from Kinmokusei It may contain only an extract, may contain only acteoside, or may contain a mixture of an extract from cinnamon and acteoside. In the case where a mixture of an extract from cinnamon and acteoside is included as an active ingredient, the blending ratio may be appropriately determined according to the degree of action of the extract from antelope and acteoside.
ここで、キンモクセイからの抽出物又はアクテオシドが有する抗肥満作用は、例えば、サイクリックＡＭＰホスホジエステラーゼ阻害作用に基づいて発揮される。ただし、キンモクセイからの抽出物又はアクテオシドが有する抗肥満作用は、上記作用に基づいて発揮される抗肥満作用に限定されるものではない。 Here, the anti-obesity action possessed by the extract or acteoside from Buttercup is exhibited based on, for example, a cyclic AMP phosphodiesterase inhibitory action. However, the anti-obesity action of the extract from Snapper or Acteoside is not limited to the anti-obesity action exhibited based on the above action.
本発明のマトリックスメタロプロテアーゼ−１阻害剤、マトリックスメタロプロテアーゼ−２阻害剤、エストロゲン様作用剤、プロフィラグリン産生促進剤、フィラグリン産生促進剤、抗肥満剤又はサイクリックＡＭＰホスホジエステラーゼ阻害剤は、キンモクセイからの抽出物及び／又はアクテオシドのみからなるものであってもよいし、上記抽出物及び／又はアクテオシドを製剤化したものであってもよい。 Matrix metalloprotease-1 inhibitor, matrix metalloproteinase-2 inhibitor, estrogen-like agent, profilagrin production promoter, filaggrin production promoter, anti-obesity agent or cyclic AMP phosphodiesterase inhibitor of the present invention It may consist only of an extract and / or acteoside, or may be a formulation of the extract and / or acteoside.
キンモクセイからの抽出物及び／又はアクテオシドは、デキストリン、シクロデキストリン等の薬学的に許容し得るキャリアーその他任意の助剤を用いて、常法に従い、粉末状、顆粒状、錠剤状、液状等の任意の剤形に製剤化することができる。この際、助剤としては、例えば、賦形剤、結合剤、崩壊剤、滑沢剤、安定剤、矯臭剤等を用いることができる。また、キンモクセイからの抽出物及び／又はアクテオシドは、他の組成物（例えば、飲食品等）に配合して使用することができる。 The extract from cinnamon moss and / or acteoside may be any powder, granule, tablet, liquid, etc., using a pharmaceutically acceptable carrier such as dextrin and cyclodextrin and other optional auxiliaries according to conventional methods. It can be formulated into a dosage form. In this case, as an auxiliary agent, for example, an excipient, a binder, a disintegrant, a lubricant, a stabilizer, a flavoring agent and the like can be used. Moreover, the extract and / or acteoside from Buttercups can be used by blending with other compositions (for example, food and drink).
なお、本発明のマトリックスメタロプロテアーゼ−１阻害剤、マトリックスメタロプロテアーゼ−２阻害剤、エストロゲン様作用剤、プロフィラグリン産生促進剤、フィラグリン産生促進剤、抗肥満剤又はサイクリックＡＭＰホスホジエステラーゼ阻害剤は、必要に応じて、マトリックスメタロプロテアーゼ−１阻害作用、マトリックスメタロプロテアーゼ−２阻害作用、エストロゲン様作用、プロフィラグリン産生促進作用、フィラグリン産生促進作用、抗肥満作用又はサイクリックＡＭＰホスホジエステラーゼ阻害作用を有する他の天然抽出物等を配合して有効成分として用いることができる。 In addition, the matrix metalloproteinase-1 inhibitor, matrix metalloproteinase-2 inhibitor, estrogen-like agent, profilagrin production promoter, filaggrin production promoter, anti-obesity agent or cyclic AMP phosphodiesterase inhibitor of the present invention are necessary. Depending on the matrix metalloproteinase-1 inhibitory action, matrix metalloprotease-2 inhibitory action, estrogen-like action, profilagulin production promoting action, filaggrin production promoting action, anti-obesity action or cyclic AMP phosphodiesterase inhibitory action An extract or the like can be blended and used as an active ingredient.
本発明のマトリックスメタロプロテアーゼ−１阻害剤、マトリックスメタロプロテアーゼ−２阻害剤、エストロゲン様作用剤、プロフィラグリン産生促進剤、フィラグリン産生促進剤、抗肥満剤又はサイクリックＡＭＰホスホジエステラーゼ阻害剤の投与方法としては、一般に経口投与が挙げられるが、疾患の種類に応じて、その予防・治療等に好適な方法を適宜選択すればよい。 As a method of administering the matrix metalloproteinase-1 inhibitor, matrix metalloproteinase-2 inhibitor, estrogen-like agent, profilagrin production promoter, filaggrin production promoter, anti-obesity agent or cyclic AMP phosphodiesterase inhibitor of the present invention In general, oral administration can be mentioned, but a suitable method for the prevention / treatment or the like may be appropriately selected depending on the type of the disease.
また、本発明のマトリックスメタロプロテアーゼ−１阻害剤、マトリックスメタロプロテアーゼ−２阻害剤、エストロゲン様作用剤、プロフィラグリン産生促進剤、フィラグリン産生促進剤、抗肥満剤又はサイクリックＡＭＰホスホジエステラーゼ阻害剤の投与量も、疾患の種類、重症度、患者の個人差、投与方法、投与期間等によって適宜増減すればよい。 The dose of the matrix metalloproteinase-1 inhibitor, matrix metalloproteinase-2 inhibitor, estrogen-like agent, profilagrin production promoter, filaggrin production promoter, anti-obesity agent or cyclic AMP phosphodiesterase inhibitor of the present invention In addition, it may be appropriately increased or decreased depending on the type of disease, severity, individual differences among patients, administration method, administration period, and the like.
本発明のマトリックスメタロプロテアーゼ−１阻害剤は、キンモクセイからの抽出物及び／又はアクテオシドが有するマトリックスメタロプロテアーゼ−１阻害作用を通じて、皮膚の老化症状を予防・改善することができるとともに、例えば、癌細胞の浸潤、転移等をも予防することができる。ただし、本発明のマトリックスメタロプロテアーゼ−１阻害剤は、これらの用途以外にもマトリックスメタロプロテアーゼ−１阻害作用を発揮することに意義のあるすべての用途に用いることができる。 The matrix metalloproteinase-1 inhibitor of the present invention can prevent and ameliorate skin aging symptoms through the matrix metalloproteinase-1 inhibitory action of an extract from Buttercup and / or acteoside, for example, cancer cells Infiltration, metastasis, etc. can be prevented. However, the matrix metalloprotease-1 inhibitor of the present invention can be used for all purposes other than these uses that are meaningful for exhibiting the matrix metalloproteinase-1 inhibitory action.
本発明のマトリックスメタロプロテアーゼ−２阻害剤は、キンモクセイからの抽出物及び／又はアクテオシドが有するマトリックスメタロプロテアーゼ−２阻害作用を通じて、皮膚の老化症状を予防・改善することができるとともに、血管新生を抑制し、腫瘍細胞への血流を抑制し、腫瘍細胞の増殖を抑制することができ、マトリックスメタロプロテアーゼ−２の発現上昇に起因する疾患（例えば、悪性腫瘍、がん等）を予防、治療又は改善することができる。ただし、本発明のマトリックスメタロプロテアーゼ−２阻害剤は、これらの用途以外にもマトリックスメタロプロテアーゼ−２阻害作用を発揮することに意義のあるすべての用途に用いることができる。 The matrix metalloproteinase-2 inhibitor of the present invention can prevent and ameliorate skin aging symptoms and inhibit angiogenesis through the matrix metalloproteinase-2 inhibitory action of extracts from cynomolgus and / or acteoside. In addition, the blood flow to tumor cells can be suppressed, the proliferation of tumor cells can be suppressed, and diseases (eg, malignant tumors, cancers, etc.) caused by increased expression of matrix metalloproteinase-2 can be prevented, treated or Can be improved. However, the matrix metalloprotease-2 inhibitor of the present invention can be used for all purposes that are meaningful in exhibiting the matrix metalloprotease-2 inhibitory action in addition to these uses.
本発明のエストロゲン様作用剤は、キンモクセイからの抽出物及び／又はアクテオシドが有するエストロゲン様作用を通じて、皮膚の老化症状を予防・改善することができるとともに、エストロゲンの欠乏に起因する疾患（例えば、冠動脈性心臓疾患（心筋梗塞等）、骨粗鬆症等）を予防、治療又は改善することができる。ただし、本発明のエストロゲン様作用剤は、これらの用途以外にもエストロゲン様作用を発揮することに意義のあるすべての用途に用いることができる。 The estrogen-like agent of the present invention can prevent and ameliorate skin aging symptoms through estrogen-like action possessed by an extract and / or acteoside from Buttercup, and diseases caused by deficiency of estrogen (for example, coronary arteries) Can prevent, treat or ameliorate congenital heart diseases (such as myocardial infarction) and osteoporosis). However, the estrogen-like agent of the present invention can be used for all uses other than these uses that are meaningful for exerting an estrogen-like action.
本発明のプロフィラグリン産生促進剤は、キンモクセイからの抽出物及び／又はアクテオシドが有するプロフィラグリン産生促進作用を通じて、細胞内でのプロフィラグリンの産生を促進し、プロフィラグリンの加水分解により得られるフィラグリン量を増加させ、皮膚の保湿能力を改善することができ、これにより、皮膚の弾力性を維持し、皮膚の老化、乾燥肌、しわ、肌荒れ等を予防・改善することができる。ただし、本発明のプロフィラグリン産生促進剤は、これらの用途以外にもプロフィラグリン産生促進作用を発揮することに意義のあるすべての用途に用いることができる。 The profilaggrin production promoter of the present invention promotes the production of profilaggrin in the cell through the profilaggrin production promoting action of the extract from cynomolgus and / or acteoside, and the amount of filaggrin obtained by hydrolysis of profilagrin , And the moisture retention ability of the skin can be improved, whereby the elasticity of the skin can be maintained, and skin aging, dry skin, wrinkles, rough skin, etc. can be prevented and improved. However, the profilaggrin production promoter of the present invention can be used for all purposes that are meaningful for exerting a profilagrin production promoting action in addition to these uses.
本発明のフィラグリン産生促進剤は、キンモクセイからの抽出物及び／又はアクテオシドが有するフィラグリン産生促進作用を通じて、細胞内でのフィラグリンの産生を促進し、皮膚の保湿能力を改善することができ、これにより、皮膚の弾力性を維持し、皮膚の老化、乾燥肌、しわ、肌荒れ等を予防・改善することができる。ただし、本発明のフィラグリン産生促進剤は、これらの用途以外にもフィラグリン産生促進作用を発揮することに意義のあるすべての用途に用いることができる。 The filaggrin production promoter of the present invention can promote the production of filaggrin in cells through the filaggrin production promoting action possessed by the extract and / or acteoside from Buttercups, thereby improving the moisturizing ability of the skin. It can maintain the elasticity of the skin and prevent / improve skin aging, dry skin, wrinkles, rough skin and the like. However, the filaggrin production promoter of the present invention can be used for all purposes that are meaningful for exerting the filaggrin production promoting action in addition to these uses.
本発明の抗肥満剤は、キンモクセイからの抽出物及び／又はアクテオシドが有するサイクリックＡＭＰホスホジエステラーゼ阻害作用を通じて、サイクリックＡＭＰの産生を促進し、脂肪細胞の分解をすることができ、この結果、肥満症を改善することができる。ただし、本発明の抗肥満剤は、これらの用途以外にもサイクリックＡＭＰホスホジエステラーゼ阻害作用を発揮することに意義のあるすべての用途に用いることができる。 The anti-obesity agent of the present invention can promote the production of cyclic AMP and decompose adipocytes through the cyclic AMP phosphodiesterase inhibitory action of the extract from cynomolgus and / or acteoside. Can improve symptoms. However, the anti-obesity agent of the present invention can be used for all purposes other than these uses that are meaningful for exerting a cyclic AMP phosphodiesterase inhibitory action.
本発明のサイクリックＡＭＰホスホジエステラーゼ阻害剤は、キンモクセイからの抽出物及び／又はアクテオシドが有するサイクリックＡＭＰホスホジエステラーゼ阻害作用を通じて、サイクリックＡＭＰの産生を促進し、脂肪細胞の分解を促進することができ、この結果、肥満症を改善することができる。ただし、本発明のサイクリックＡＭＰホスホジエステラーゼ阻害剤は、これらの用途以外にもサイクリックＡＭＰホスホジエステラーゼ阻害作用を発揮することに意義のあるすべての用途に用いることができる。 The cyclic AMP phosphodiesterase inhibitor of the present invention can promote cyclic AMP production and promote adipocyte degradation through the cyclic AMP phosphodiesterase inhibitory action possessed by the extract and / or acteoside from Buttercups, As a result, obesity can be improved. However, the cyclic AMP phosphodiesterase inhibitor of the present invention can be used for all purposes other than these uses that are meaningful for exerting a cyclic AMP phosphodiesterase inhibitory action.
なお、本発明のマトリックスメタロプロテアーゼ−１阻害剤、マトリックスメタロプロテアーゼ−２阻害剤、エストロゲン様作用剤、プロフィラグリン産生促進剤、フィラグリン産生促進剤、抗肥満剤又はサイクリックＡＭＰホスホジエステラーゼ阻害剤は、ヒトに対して好適に適用されるものであるが、それぞれの作用効果が奏される限り、ヒト以外の動物に対して適用することもできる。 The matrix metalloproteinase-1 inhibitor, matrix metalloproteinase-2 inhibitor, estrogen-like agent, profilagrin production promoter, filaggrin production promoter, anti-obesity agent or cyclic AMP phosphodiesterase inhibitor of the present invention is human. However, it can also be applied to animals other than humans as long as each effect is exhibited.
以下、製造例及び試験例を示し、本発明を具体的に説明するが、本発明は下記の各例に何ら制限されるものではない。 Hereinafter, although a manufacture example and a test example are shown and this invention is demonstrated concretely, this invention is not restrict | limited to each following example at all.
〔製造例１〕キンモクセイ花部抽出物の製造
キンモクセイの花部の粗粉砕物１５０ｇに５０質量％エタノール（水とエタノールとの質量比＝１：１）１５００ｍＬを加え、穏やかに攪拌しながら８０℃にて２時間保ち、熱時濾過した。続いて、残渣に５０質量％エタノール１５００ｍＬを加え、穏やかに攪拌しながら８０℃にて２時間保ち、熱時濾過した。得られた濾液を合わせて減圧下にて濃縮し、減圧乾燥機で乾燥してキンモクセイ花部抽出物（７１．４ｇ，試料１）を得た。
[Production Example 1] Manufacture of an extract of flower portion of cinnamomum 1500 ml of 50% by weight ethanol (mass ratio of water to ethanol = 1: 1) is added to 150 g of a roughly pulverized flower portion of an antrum, and the mixture is gently stirred at 80 ° C. And filtered for 2 hours. Subsequently, 1500 mL of 50% by mass ethanol was added to the residue, kept at 80 ° C. for 2 hours with gentle stirring, and filtered while hot. The obtained filtrates were combined, concentrated under reduced pressure, and dried with a vacuum dryer to obtain an extract of flowering florets (71.4 g, sample 1).
〔製造例２〕アクテオシドの製造
製造例１により得られたキンモクセイ花部抽出物（試料１）８．９ｇをクロロホルム／メタノール／水＝１０：５：１の混合溶液に溶解し、シリカゲル（商品名：シリカゲル６０，メルク社製）を充填したガラス製のカラム上部より流入して、シリカゲルに吸着させた。ガラス製のカラムに移動層としてクロロホルム／メタノール／水＝１０：５：１を流し、その溶出液を集め、溶媒を留去して、精製物（２．５ｇ，試料２）を得た。
[Production Example 2] Manufacture of Acteoside 8.9 g of the flower extract (sample 1) obtained from Production Example 1 was dissolved in a mixed solution of chloroform / methanol / water = 10: 5: 1 and silica gel (trade name) : Silica gel 60, manufactured by Merck & Co., Inc.) and was adsorbed on silica gel. Chloroform / methanol / water = 10: 5: 1 was passed as a moving bed through a glass column, the eluate was collected, and the solvent was distilled off to obtain a purified product (2.5 g, sample 2).
上記のようにして精製して得られた精製物について、マススペクトル分析、１Ｈ−ＮＭＲ分析及び１３Ｃ−ＮＭＲ分析をした。かかる分析結果を下記に示す。 The purified product obtained by purification as described above was subjected to mass spectral analysis, 1 H-NMR analysis and 13 C-NMR analysis. The analysis results are shown below.
＜マススペクトル（ＥＳＩ−ＭＳ）＞
[Ｍ−Ｈ]− m/z 623（理論値：Ｃ２９Ｈ３６Ｏ１５−Ｈ＝623）
[Ｍ＋Ｎａ]＋ m/z 647（理論値：Ｃ２９Ｈ３６Ｏ１５＋Ｎａ＝647）
<Mass spectrum (ESI-MS)>
[M-H] - m / z 623 ( theoretical value: C 29 H 36 O 15 -H = 623)
[M + Na] + m / z 647 (theoretical value: C 29 H 36 O 15 + Na = 647)
＜１Ｈ−ＮＭＲケミカルシフトδ（帰属水素）：＞
6.69 (1H, d, J=2.0 Hz, H-2), 6.67 (1H, d, J=8.1 Hz, H-5), 6.55 (1H, dd, J=2.0, 8.1Hz, H-6), 2.78 (2H, t-like, H-7), 4.01 (1H, overlapped, H-8a), 3.70 (1H, overlapped, H-8b), 7.05 (1H, d, J=2.2 Hz, H-2’), 6.77 (1H, d, J=8.3 Hz, H-5’), 6.94 (1H, dd, J=2.2, 8.3Hz, H-6’), 7.58 (1H, d, J=15.9 Hz, H-7’), 6.27 (1H, d, J=15.9 Hz, H-8’), 4.36 (1H, d, J=7.8Hz, GlcH-1), 3.39 (1H, dd, J=7.8, 8.1Hz, GlcH-2), 3.80(1H, br.t, J=9.0 Hz, Glc H-3), 4.88 (1H, br.t, J=9.2 Hz, Glc H-4), 3.58 (1H, overlapped, Glc H-5), 3.48 (2H, overlapped, Glc H-6), 5.18( 1H, br.s, Rha H-1), 3.90 (1H, m, Rha H-2), 3.58 (2H, overlapped, Rha-H-3 and Rha H-5), 3.30(1H, overlapped, Rha-H-4), 1.08.(3H, d, J=6.1 Hz, Rha H-6)
< 1 H-NMR chemical shift δ (assigned hydrogen):>
6.69 (1H, d, J = 2.0 Hz, H-2), 6.67 (1H, d, J = 8.1 Hz, H-5), 6.55 (1H, dd, J = 2.0, 8.1Hz, H-6), 2.78 (2H, t-like, H-7), 4.01 (1H, overlapped, H-8a), 3.70 (1H, overlapped, H-8b), 7.05 (1H, d, J = 2.2 Hz, H-2 ' ), 6.77 (1H, d, J = 8.3 Hz, H-5 '), 6.94 (1H, dd, J = 2.2, 8.3Hz, H-6'), 7.58 (1H, d, J = 15.9 Hz, H -7 '), 6.27 (1H, d, J = 15.9 Hz, H-8'), 4.36 (1H, d, J = 7.8Hz, GlcH-1), 3.39 (1H, dd, J = 7.8, 8.1Hz , GlcH-2), 3.80 (1H, br.t, J = 9.0 Hz, Glc H-3), 4.88 (1H, br.t, J = 9.2 Hz, Glc H-4), 3.58 (1H, overlapped, Glc H-5), 3.48 (2H, overlapped, Glc H-6), 5.18 (1H, br.s, Rha H-1), 3.90 (1H, m, Rha H-2), 3.58 (2H, overlapped, Rha-H-3 and Rha H-5), 3.30 (1H, overlapped, Rha-H-4), 1.08. (3H, d, J = 6.1 Hz, Rha H-6)
＜１３Ｃ−ＮＭＲケミカルシフトδ（帰属炭素）：＞
131.1(s, C-1), 116.1(d, C-2), 146.5(s, C-3), 144.3(s, C-4), 116.8(d, C-5), 121.0(d, C-6), 36.4(t, C-7), 72.0(t, C-8), 127.3(s, C-1’), 114.9(d, C-2’), 145.8(s, C-3’), 149.4(s, C-4’), 116.2(d, C-5’), 122.9(d, C-6’), 147.7(d, C-7’), 114.4(d, C-8’), 167.9(s, C-9’), 103.9(d, Glc C-1), 75.9(d, Glc C-2), 81.4(d, Glc C-3), 70.7(d, Glc C-4), 75.8(d, Glc C-5), 62.1(t, Glc C-6), 102.7(d, Rha C-1), 72.1(d, Rha C-2), 71.8(d, Rha C-3), 73.5(d, Rha C-4), 70.2(d, RhaC-5), 18.3(q, Rha C-6)
< 13 C-NMR chemical shift δ (assigned carbon):>
131.1 (s, C-1), 116.1 (d, C-2), 146.5 (s, C-3), 144.3 (s, C-4), 116.8 (d, C-5), 121.0 (d, C -6), 36.4 (t, C-7), 72.0 (t, C-8), 127.3 (s, C-1 '), 114.9 (d, C-2'), 145.8 (s, C-3 ' ), 149.4 (s, C-4 '), 116.2 (d, C-5'), 122.9 (d, C-6 '), 147.7 (d, C-7'), 114.4 (d, C-8 ' ), 167.9 (s, C-9 '), 103.9 (d, Glc C-1), 75.9 (d, Glc C-2), 81.4 (d, Glc C-3), 70.7 (d, Glc C-4 ), 75.8 (d, Glc C-5), 62.1 (t, Glc C-6), 102.7 (d, Rha C-1), 72.1 (d, Rha C-2), 71.8 (d, Rha C-3 ), 73.5 (d, Rha C-4), 70.2 (d, RhaC-5), 18.3 (q, Rha C-6)
以上の分析結果から、キンモクセイ花部抽出物から得られた化合物が、下記式（Ｉ）で表されるアクテオシド（Acteoside）であることが確認された。 From the above analysis results, it was confirmed that the compound obtained from the extract of the flowering part of the beetle was Acteoside represented by the following formula (I).
〔試験例１〕マトリックスメタロプロテアーゼ−１（ＭＭＰ−１）阻害作用試験
製造例１にて得られたキンモクセイ花部抽出物（試料１）及び製造例２にて得られたアクテオシド（Acteoside，試料２）について、以下のようにして間オリックスメタロプロテアーゼ−１（ＭＭＰ−１）阻害作用を試験した。
[Test Example 1] Matrix metalloprotease-1 (MMP-1) inhibitory action test Kintosei flower part extract obtained in Production Example 1 (Sample 1) and Acteoside obtained in Production Example 2 (Acteoside, Sample 2) ) Was tested for interorix metalloprotease-1 (MMP-1) inhibitory action as follows.
蓋付試験管にて２０ｍｍｏｌ／Ｌの塩化カルシウムを含有する０．１ｍｏｌ／ＬのＴｒｉｓ−ＨＣＬ緩衝液（ｐＨ７．１）に溶解した試料溶液（試料濃度は表１を参照）５０μＬ、ＭＭＰ−１溶液（COLLAGENASE Type IV from Clostridium histolyticum，Sigma社製）５０μＬ及びPz-peptide溶液（Pz-Pro-Leu-Gly-Pro-D-Arg-OH，BACHEM Feinchemikalien AG社製）４００μＬを混合し、３７℃にて３０分反応させた後、２５ｍｍｏｌ／Ｌのクエン酸溶液１ｍＬを加え反応を停止した。その後、酢酸エチル５ｍＬを加え、激しく振とうした。これを遠心（１６００×ｇ，１０分）し、酢酸エチル層の波長２３０ｎｍにおける吸光度を測定した。また、同様にして空試験を行い補正した。得られた結果から、下記式により、ＭＭＰ−１阻害率（％）を算出した。 Sample solution (see Table 1 for sample concentration) dissolved in 0.1 mol / L Tris-HCL buffer (pH 7.1) containing 20 mmol / L calcium chloride in a test tube with a lid, MMP-1 50 μL of the solution (COLLAGENASE Type IV from Clostridium histolyticum, manufactured by Sigma) and 400 μL of the Pz-peptide solution (Pz-Pro-Leu-Gly-Pro-D-Arg-OH, manufactured by BACHEM Feinchemikalien AG) were mixed at 37 ° C. Then, the reaction was stopped by adding 1 mL of a 25 mmol / L citric acid solution. Thereafter, 5 mL of ethyl acetate was added and shaken vigorously. This was centrifuged (1600 × g, 10 minutes), and the absorbance of the ethyl acetate layer at a wavelength of 230 nm was measured. Similarly, a blank test was performed and corrected. From the obtained results, the MMP-1 inhibition rate (%) was calculated by the following formula.
ＭＭＰ−１阻害率(％)={１−(Ｃ−Ｄ)／(Ａ−Ｂ)}×１００
式中、Ａは「試料無添加、酵素添加での波長３２０ｎｍにおける吸光度」を表し、Ｂは「試料無添加、酵素無添加での波長３２０ｎｍにおける吸光度」を表し、Ｃは「試料添加、酵素添加での波長３２０ｎｍにおける吸光度」を表し、Ｄは、「試料添加、酵素無添加での波長３２０ｎｍにおける吸光度」を表す。
MMP-1 inhibition rate (%) = {1- (C−D) / (A−B)} × 100
In the formula, A represents “absorbance at a wavelength of 320 nm when no sample is added and an enzyme is added”, B represents “absorbance at a wavelength of 320 nm when no sample is added and an enzyme is not added”, and C is “addition of sample and enzyme is added” And “D” represents “absorbance at a wavelength of 320 nm when a sample is added and no enzyme is added”.
また、試料濃度を段階的に減少させて上記ＭＭＰ−１阻害率の測定を行い、ＭＭＰ−１阻害率が５０％になる試料濃度ＩＣ５０（μｇ／ｍＬ）を内挿法により求めた。
結果を表１に示す。
Further, the MMP-1 inhibition rate was measured by gradually reducing the sample concentration, and the sample concentration IC 50 (μg / mL) at which the MMP-1 inhibition rate was 50% was determined by interpolation.
The results are shown in Table 1.
表１に示すように、キンモクセイ花部抽出物及びアクテオシドは、ＭＭＰ−１阻害作用を有することが確認された。また、キンモクセイ花部抽出物及びアクテオシドのＭＭＰ−１阻害作用の程度は、キンモクセイ花部抽出物及びアクテオシドの濃度により調節可能であることが確認された。 As shown in Table 1, it was confirmed that the extract of the flowering quince flower part and acteoside has an MMP-1 inhibitory action. In addition, it was confirmed that the degree of the MMP-1 inhibitory action of the flower extract and acteoside can be adjusted by the concentration of the flower extract and acteoside.
〔試験例２〕マトリックスメタロプロテアーゼ−２（ＭＭＰ−２）阻害作用試験
製造例１にて得られたキンモクセイ花部抽出物（試料１）及び製造例２にて得られたアクテオシド（Acteoside，試料２）について、以下のようにしてマトリックスメタロプロテアーゼ−２（ＭＭＰ−２）阻害作用を試験した。
[Test Example 2] Matrix metalloprotease-2 (MMP-2) inhibitory action test Kintosei flower part extract obtained in Production Example 1 (Sample 1) and Acteoside obtained in Production Example 2 (Acteoside, Sample 2) ) Was tested for matrix metalloproteinase-2 (MMP-2) inhibitory action as follows.
（１）ＭＭＰ−２の調製
ヒトＭＭＰ−２ ｃＤＮＡを、下記の５'ＰＣＲプライマー及び３'ＰＣＲプライマーを用い、ＭＭＰ−２の３．３ｋｂ ｃＤＮＡ断片を含むｐＳＧ−ＧｅｌＡを鋳型としてＰＣＲ反応を行った。得られた３０Ａｌａから４７４Ｖａｌまでをコードする１．３ｋｂ ＰＣＲ断片をＢａｍ Ｈｌ／Ｓａｌｌで消化し、ｐＴＨ−７２発現ベクター（ｐＴＨ−ＭＭＰ２−ＰＣ）のＢａｍ Ｈｌ／Ｓａｌｌ部位にクローン化した。
５'ＰＣＲプライマー：5'-ggcggatccatggcgccgtcgcccatcatc-3'
３'ＰＣＲプライマー：3'-gccgtcgactacaatgtcctgtttgcagat-5'
(1) Preparation of MMP-2 A human MMP-2 cDNA was subjected to a PCR reaction using the following 5 ′ PCR primer and 3 ′ PCR primer, and pSG-GelA containing a 3.3 kb cDNA fragment of MMP-2 as a template. It was. The resulting 1.3 kb PCR fragment encoding from 30 Ala to 474 Val was digested with Bam H1 / Sall and cloned into the Bam H1 / Sall site of the pTH-72 expression vector (pTH-MMP2-PC).
5 'PCR primer: 5'-ggcggatccatggcgccgtcgcccatcatc-3'
3 'PCR primer: 3'-gccgtcgactacaatgtcctgtttgcagat-5'
ヒトＭＭＰ−２の発現は、「Itoh, M. et al.; J. Biolchem., Vol.119, pp667-673, 1996」に準じて、精製及び巻き戻し（以下「リフォールディング」という。）は、「西村義文、大野茂雄 監修，タンパク実験プロトコール 細胞工学別冊 実験プロトコールシリーズ２ 構造解析編，１９９７」に準じて行った。また、ｃＤＮＡは、「Collier, I. E. et al., J. Biol. Chem., Vol.263, No.14, pp6579-6587, 1998」に記載されているものを使用した。 The expression of human MMP-2 was purified and unwound (hereinafter referred to as “refolding”) according to “Itoh, M. et al .; J. Biolchem., Vol.119, pp667-673, 1996”. , “Supervised by Yoshifumi Nishimura, Shigeo Ohno, Protein Experiment Protocol, Cell Engineering Separate Volume, Experiment Protocol Series 2 Structural Analysis, 1997”. The cDNA described in "Collier, IE et al., J. Biol. Chem., Vol. 263, No. 14, pp6579-6587, 1998" was used.
すなわち、ＭＭＰ−２のｃＤＮＡを含む発現ベクター（ｐＴＨ−ＭＭＰ−２）を、大腸菌ＢＬ２１（ＤＥ３）株にトランスフェクトし、ＩＰＴＧで発現誘導した。発現タンパクは、Ｎｉ−ＮＴＡ樹脂（QUIAGEN INC.社製）を用いてアフィニティー精製後、リフォールディングを行い、酢酸４−アミノフェニル水銀と３７℃で６０分間反応を行うことで活性型へ移行させた後、ＥＤＴＡを加えた。これを酵素標本とし、蛍光性ペプチド基質（MOCAc/DNP peptide）切断活性反応を行い、高速液体クロマトグラフィー（カラム：Wakosil 5C18，溶離液：３０％アセトニトリル＋０．１％ＴＨＦ，流速：１．０ｍＬ／ｍｉｎ，検出：励起波長３２５ｎｍ，蛍光波長４１０ｎｍ）による生成物のピーク面積を測定し、これを酵素活性の指標とした。 That is, an expression vector (pTH-MMP-2) containing MMP-2 cDNA was transfected into Escherichia coli BL21 (DE3) strain, and expression was induced with IPTG. The expressed protein was affinity-purified using Ni-NTA resin (manufactured by QUIAGEN INC.), Refolded, and reacted with 4-aminophenylmercuric acetate at 37 ° C. for 60 minutes to shift to the active form. Later, EDTA was added. Using this as an enzyme sample, a fluorescent peptide substrate (MOCAc / DNP peptide) cleavage activity reaction was performed, and high performance liquid chromatography (column: Wakosil 5C18, eluent: 30% acetonitrile + 0.1% THF, flow rate: 1.0 mL / min, detection: excitation wavelength 325 nm, fluorescence wavelength 410 nm) was measured, and this was used as an index of enzyme activity.
（２）ＭＭＰ−２阻害活性の測定
試料を蒸留水に溶解させて８．０ｍｇ／ｍＬとした後、蒸留水にて４．０ｍｇ／ｍＬ、２．０ｍｇ／ｍＬに希釈し、懸濁物を除くため濾過した。ＭＭＰ−２阻害活性の測定は、活性型ＭＭＰ−２ ４０μＬ、試料溶液（試料濃度は表２を参照）２０μＬ、アッセイバッファー（５００ｍＭのトリス−塩酸緩衝液（ｐＨ７．５）、１．５Ｍの塩化ナトリウム、１００ｍＭの塩化カルシウム、５００μＭの硫酸亜鉛、３０ｍＭのアジ化ナトリウム、０．０５％のＢｒｉｊ３５）２０μＬを、３７℃で１５分間プレインキュベーションした後、MOCAc/DNP peptide１２０μＬ（４．１６μＭ）を添加し、３７℃で２時間反応させ、その後ＥＤＴＡ１０μＬ（２００ｍＭ）を添加した。反応液中の生成物について高速液体クロマトグラフィー分析によるピーク面積を測定した。試料溶液の代わりに蒸留水を加えた反応液の生成物を１００％として試料のＭＭＰ−２阻害率（％）を算出した。
(2) Measurement of MMP-2 inhibitory activity The sample was dissolved in distilled water to 8.0 mg / mL, diluted with distilled water to 4.0 mg / mL and 2.0 mg / mL, and the suspension was Filtered to remove. MMP-2 inhibitory activity was measured by 40 μL of active MMP-2, 20 μL of sample solution (see Table 2 for sample concentration), assay buffer (500 mM Tris-HCl buffer (pH 7.5), 1.5 M chloride) 20 μL of sodium, 100 mM calcium chloride, 500 μM zinc sulfate, 30 mM sodium azide, 0.05% Brij 35) is preincubated for 15 minutes at 37 ° C., and then 120 μL of MOCAc / DNP peptide (4.16 μM) is added. The mixture was reacted at 37 ° C. for 2 hours, and then 10 μL of EDTA (200 mM) was added. The peak area by the high performance liquid chromatography analysis was measured about the product in a reaction liquid. The MMP-2 inhibition rate (%) of the sample was calculated with the product of the reaction solution with distilled water added instead of the sample solution as 100%.
また、試料濃度を段階的に減少させて上記ＭＭＰ−２阻害率の測定を行い、ＭＭＰ−２阻害率が５０％になる試料濃度ＩＣ５０（μｇ／ｍＬ）を内挿法により求めた。
結果を表２に示す。
Further, the MMP-2 inhibition rate was measured by gradually reducing the sample concentration, and the sample concentration IC 50 (μg / mL) at which the MMP-2 inhibition rate was 50% was determined by interpolation.
The results are shown in Table 2.
表２に示すように、キンモクセイ花部抽出物及びアクテオシドは、ＭＭＰ−２阻害作用を有することが確認された。また、キンモクセイ花部抽出物及びアクテオシドのＭＭＰ−２阻害作用の程度は、キンモクセイ花部抽出物及びアクテオシドの濃度により調節可能であることが確認された。 As shown in Table 2, it was confirmed that the extract of the flowering quince flower part and acteoside has an MMP-2 inhibitory action. In addition, it was confirmed that the degree of the MMP-2 inhibitory action of the beetle flower part extract and acteoside can be adjusted by the concentration of the beetle flower part extract and acteoside.
〔試験例３〕エストロゲン様作用試験
製造例１にて得られたキンモクセイ花部抽出物（試料１）及び製造例２にて得られたアクテオシド（Acteoside，試料２）について、以下のようにしてエストロゲン様作用を試験した。
[Test Example 3] Estrogen-like action test The estrogen was extracted from the flower extract (sample 1) obtained in Production Example 1 and the acteoside obtained in Production Example 2 as follows. The effect was tested.
ヒト乳癌由来細胞（ＭＣＦ−７）を１０％ＦＢＳ、１％ＮＥＡＡ及び１ｍｍｏｌ／Ｌのピルビン酸ナトリウムを含有するＭＥＭを用いて培養した後、トリプシン処理により細胞を回収した。回収した細胞を、活性炭処理した１０％ＦＢＳ、１％ＮＥＡＡ及び１ｍｍｏｌ／Ｌのピルビン酸ナトリウムを含有しフェノールレッドを含有しないＭＥＭ（Ｔ−ＭＥＭ）を用いて、３．０×１０４ｃｅｌｌｓ／ｍＬの細胞密度に調整した後、４８ウェルプレートに１ウェルあたり４５０μＬずつ播種し、細胞を定着させるため培養した。６時間後（０日目）にＴ−ＭＥＭ培地で終濃度の１０倍に調整した試料溶液（試料濃度は表３を参照）を各ウェルに５０μＬずつ添加し培養を続けた。３日目に培地を抜き、Ｔ−ＭＥＭで終濃度に調整した試料溶液を各ウェルに０．５ｍＬずつ添加し、さらに培養を続けた。 Human breast cancer-derived cells (MCF-7) were cultured using MEM containing 10% FBS, 1% NEAA and 1 mmol / L sodium pyruvate, and then cells were collected by trypsin treatment. The collected cells were treated with activated carbon-treated 10% FBS, 1% NEAA, and 1 mmol / L sodium pyruvate and no phenol red (T-MEM) at 3.0 × 10 4 cells / mL. The cell density was adjusted to 450 μL per well in a 48-well plate and cultured to establish the cells. Six hours later (day 0), 50 μL of a sample solution adjusted to 10 times the final concentration in T-MEM medium (see Table 3 for the sample concentration) was added to each well, and the culture was continued. On the third day, the medium was removed, 0.5 mL of the sample solution adjusted to the final concentration with T-MEM was added to each well, and the culture was further continued.
エストロゲン様作用は、ＭＴＴアッセイ法を用いて測定した。培養終了後、培地を抜き、１％ＮＥＡＡ及び１ｍｍｏｌ／Ｌのピルビン酸ナトリウムを含有するＭＥＭに終濃度０．４ｍｇ／ｍＬで溶解したＭＴＴを各ウェルに２００μＬずつ添加した。２時間培養した後に、細胞内に生成したブルーホルマザンを２−プロパノール２００μＬで抽出した。抽出後、波長５７０ｎｍにおける吸光度を測定した。同時に濁度として波長６５０ｎｍにおける吸光度を測定し、両者の差をもってブルーホルマザン生成量とした。ポジティブコントロールとして、１×１０−９Ｍのエストラジオールを使用した。得られた結果から、下記式により、試料溶液添加時のエストロゲン様作用率（％）を算出した。 Estrogen-like effects were measured using the MTT assay. After completion of the culture, the medium was removed, and 200 μL of MTT dissolved in MEM containing 1% NEAA and 1 mmol / L sodium pyruvate at a final concentration of 0.4 mg / mL was added to each well. After culturing for 2 hours, blue formazan produced in the cells was extracted with 200 μL of 2-propanol. After extraction, the absorbance at a wavelength of 570 nm was measured. At the same time, the absorbance at a wavelength of 650 nm was measured as turbidity, and the difference between the two was used as the amount of blue formazan produced. As a positive control, 1 × 10 −9 M estradiol was used. From the obtained results, the estrogen-like action rate (%) at the time of adding the sample solution was calculated by the following formula.
エストロゲン様作用率(％)＝Ａ／Ｂ×１００
式中、Ａは「試料溶液添加時の吸光度」を表し、Ｂは「試料溶液無添加時の吸光度」を表す。
結果を表３に示す。
Estrogen-like action rate (%) = A / B × 100
In the formula, A represents “absorbance when a sample solution is added”, and B represents “absorbance when no sample solution is added”.
The results are shown in Table 3.
表３に示すように、キンモクセイ花部抽出物及びアクテオシドは、エストロゲン様作用を有することが確認された。 As shown in Table 3, it was confirmed that the extract of the floret flower part and acteoside has an estrogen-like action.
〔試験例４〕プロフィラグリン／フィラグリン産生促進作用試験
製造例１にて得られたキンモクセイ花部抽出物（試料１）及び製造例２にて得られたアクテオシド（Acteoside，試料２）について、以下のようにしてプロフィラグリン産生促進作用及びフィラグリン産生促進作用を試験した。
[Test Example 4] Profilaggrin / filaggrin production promoting action test For the extract of the floret flower part (Sample 1) obtained in Production Example 1 and Acteoside (Acteoside, Sample 2) obtained in Production Example 2, the following Thus, the profilaggrin production promoting effect and the filaggrin production promoting effect were tested.
正常ヒト新生児皮膚表皮角化細胞（ＮＨＥＫ）を８０ｃｍ２のフラスコで正常ヒト表皮角化細胞培地（ＫＧＭ）にて３７℃、５％ＣＯ２−９５％ａｉｒの条件下で培養し、常法により細胞を集めた。得られた細胞を上記培地にて１．５×１０５個／ｍＬの細胞密度に調整し、２ｍＬずつ６穴コラーゲンコートプレートに播種して３７℃、５％ＣＯ２−９５％ａｉｒの条件下にて３日間培養した。培養後、０．５％ＤＭＳＯに溶解した試料を含むＫＧＭ２ｍＬに交換し、３７℃、５％ＣＯ２−９５％ａｉｒの条件下にて５日間培養した。培養終了後、常法により総タンパクの調整を行った。 Normal human newborn skin epidermal keratinocytes (NHEK) were cultured in normal human epidermal keratinocyte medium (KGM) in an 80 cm 2 flask under conditions of 37 ° C. and 5% CO 2 -95% air. Cells were collected. The obtained cells were adjusted to a cell density of 1.5 × 10 5 cells / mL with the above medium, seeded on a 6-well collagen-coated plate in 2 mL increments at 37 ° C., 5% CO 2 -95% air. For 3 days. After culturing, the sample was exchanged for 2 mL of KGM containing a sample dissolved in 0.5% DMSO, and cultured for 5 days under conditions of 37 ° C. and 5% CO 2 -95% air. After completion of the culture, total protein was prepared by a conventional method.
＜ウェスタンブロッティング＞
１０μｇ／列に調整したサンプルをＳＤＳ−ＰＡＧＥにより展開し、ＰＶＤＦ膜に転写した。５％スキムミルクを含むＰＢＳ（−）でブロッキングを行った後、抗ヒトフィラグリンモノクローナル抗体（Harbor Bio-Products）、ビオチン標識抗マウスＩｇ（Whole Ab，Amersham Biosciences社製）及びストレプトアビジンーペルオキシダーゼ複合体（CALBIOCHEM社製）を、０．１％Ｔｗｅｅｎ２０、０．３％スキムミルクを含むＰＢＳ（−）で１０００倍に希釈して順次反応させ、ECL Western blotting detection reagents and analysis system（Amersham Biosciences社製）の発光によりプロフィラグリン及びフィラグリンを検出した。検出したバンドをKODAK 1D Image Analysis Software EDAS290 Version3.5にて定量的に測定した。
<Western blotting>
The sample adjusted to 10 μg / row was developed by SDS-PAGE and transferred to a PVDF membrane. After blocking with PBS (−) containing 5% skim milk, anti-human filaggrin monoclonal antibody (Harbor Bio-Products), biotin-labeled anti-mouse Ig (Whole Ab, manufactured by Amersham Biosciences) and streptavidin-peroxidase complex ( CALBIOCHEM) was diluted 1000-fold with PBS (-) containing 0.1% Tween20 and 0.3% skim milk, and allowed to react sequentially, and luminescence from ECL Western blotting detection reagents and analysis system (Amersham Biosciences) Was used to detect profilaggrin and filaggrin. The detected band was quantitatively measured with KODAK 1D Image Analysis Software EDAS290 Version3.5.
結果は、試料添加及び無添加で培養した細胞のそれぞれから調製したタンパク１０μｇ中のプロフィラグリン及びフィラグリンのNet intensity（バンド強度）を合算した値を用いて、試料のプロフィラグリン産生促進作用を評価し、プロフィラグリン産生促進率（％）を下記式に基づいて算出した。 As a result, the profilaggrin production promoting effect of the sample was evaluated using the value obtained by adding the net intensity (band intensity) of profilagrin and filaggrin in 10 μg of the protein prepared from each of the cells cultured with and without the sample added. The profilaggrin production promotion rate (%) was calculated based on the following formula.
プロフィラグリン産生促進率(％)＝Ａ／Ｂ×１００
上記式において、Ａは「試料添加時のNet intensity（プロフィラグリン及びフィラグリンの合計値）」を、Ｂは「試料無添加時（コントロール）のNet intensity」を表す。
結果を表４に示す。
Profilaggrin production promotion rate (%) = A / B × 100
In the above formula, A represents “Net intensity at the time of sample addition (total value of profilagrin and filaggrin)” and B represents “Net intensity at the time of no sample addition (control)”.
The results are shown in Table 4.
表４に示すように、キンモクセイ花部抽出物及びアクテオシドは、プロフィラグリン産生促進作用及びフィラグリン産生促進作用を有することが確認された。 As shown in Table 4, it was confirmed that the extract of floret flowers and acteoside has a profilagrine production promoting action and a filaggrin production promoting action.
〔試験例５〕サイクリックＡＭＰホスホジエステラーゼ阻害試験
製造例１にて得られたキンモクセイ花部抽出物（試料１）及び製造例２にて得られたアクテオシド（Acteoside，試料２）について、以下のようにしてサイクリックＡＭＰホスホジエステラーゼ阻害作用を試験した。
[Test Example 5] Cyclic AMP phosphodiesterase inhibition test For the extract of the flower part of cypress flower obtained in Production Example 1 (Sample 1) and Acteoside obtained in Production Example 2 (Acteoside, Sample 2), the following procedure was performed. The cyclic AMP phosphodiesterase inhibitory action was tested.
５ｍＭ塩化マグネシウムを含有する５０ｍＭトリス塩酸緩衝液（ｐＨ７．５）０．２ｍＬに２．５ｍｇ／ｍＬウシ血清アルブミン溶液０．１ｍＬおよび０．１ｍｇ／ｍＬサイクリックＡＭＰホスホジエステラーゼ溶液０．１ｍＬを加え、さらに試料溶液（試料濃度は表７を参照）を加え、３７℃で５分間予備反応した。これに０．５ｍｇ／ｍＬ ｃＡＭＰ溶液０．０５ｍＬを加え、３７℃で３０分間反応させた。反応後、沸騰水上で３分間煮沸して反応を停止させ、遠心（２２６０×ｇ、１０分、４℃）し、上清を試料反応液とし、下記の条件で高速液体クロマトグラフィー分析をした。また、同様の方法で空試験を行い補正した。 Add 0.2 mL of 2.5 mg / mL bovine serum albumin solution and 0.1 mL of 0.1 mg / mL cyclic AMP phosphodiesterase solution to 0.2 mL of 50 mM Tris-HCl buffer (pH 7.5) containing 5 mM magnesium chloride, and Sample solution (see Table 7 for sample concentration) was added and pre-reacted at 37 ° C. for 5 minutes. To this, 0.05 mL of a 0.5 mg / mL cAMP solution was added and reacted at 37 ° C. for 30 minutes. After the reaction, the reaction was stopped by boiling for 3 minutes on boiling water, centrifuged (2260 × g, 10 minutes, 4 ° C.), the supernatant was used as a sample reaction solution, and high performance liquid chromatography analysis was performed under the following conditions. In addition, a blank test was performed and corrected in the same manner.
＜高速液低クロマトグラフィー条件＞
Column : Wakosil C18-ODS 5μm（和光純薬工業社製）
Mobil phase : 1mM TBAP in 25mM KH2PO4 : CH3CN = 90 : 10
Flow rate : 1.0mL/min
Detector : 260nm
Atten : 32〜64
<High performance liquid low chromatography conditions>
Column: Wakosil C18-ODS 5μm (Wako Pure Chemical Industries)
Mobil phase: 1mM TBAP in 25mM KH 2 PO 4 : CH 3 CN = 90: 10
Flow rate: 1.0mL / min
Detector: 260nm
Atten: 32-64
このようにして測定されたｃＡＭＰのピーク面積から、サイクリックＡＭＰホスホジエステラーゼ阻害率（％）を下記式により算出した。 From the cAMP peak area thus measured, the cyclic AMP phosphodiesterase inhibition rate (%) was calculated by the following formula.
サイクリックＡＭＰホスホジエステラーゼ阻害率(％)＝(１−Ａ／Ｂ)×１００
式中、Ａは「試料添加時のｃＡＭＰのピーク面積」を表し、Ｂは「試料無添加時のｃＡＭＰのピーク面積」を表す。
結果を表５に示す。
Cyclic AMP phosphodiesterase inhibition rate (%) = (1−A / B) × 100
In the formula, A represents “cAMP peak area at the time of sample addition”, and B represents “cAMP peak area at the time of no sample addition”.
The results are shown in Table 5.
表５に示すように、キンモクセイ花部抽出物及びアクテオシドは、サイクリックＡＭＰホスホジエステラーゼ阻害作用を有することが確認された。また、キンモクセイ花部抽出物及びアクテオシドのサイクリックＡＭＰホスホジエステラーゼ阻害作用の程度は、キンモクセイ花部抽出物及びアクテオシドの濃度により調節可能であることが確認された。 As shown in Table 5, it was confirmed that the extract of floret flowers and acteoside has a cyclic AMP phosphodiesterase inhibitory action. In addition, it was confirmed that the degree of the inhibitory action of cynomolgus floret extract and acteoside on cyclic AMP phosphodiesterase can be adjusted by the concentration of cypress floret extract and acteoside.
本発明のマトリックスメタロプロテアーゼ−１阻害剤、マトリックスメタロプロテアーゼ−２阻害剤、エストロゲン様作用剤、プロフィラグリン産生促進剤及びフィラグリン産生促進剤は、皮膚の老化症状の予防・改善に有用であり、本発明の抗肥満剤及びサイクリックＡＭＰホスホジエステラーゼ阻害剤は、肥満症の予防・改善に有用である。 The matrix metalloproteinase-1 inhibitor, the matrix metalloproteinase-2 inhibitor, the estrogen-like agent, the profilagrin production promoter and the filaggrin production promoter of the present invention are useful for the prevention and improvement of skin aging symptoms. The anti-obesity agent and the cyclic AMP phosphodiesterase inhibitor of the invention are useful for the prevention / amelioration of obesity.
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Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2009196908A (en) * | 2008-02-20 | 2009-09-03 | Yomeishu Seizo Co Ltd | Lipase inhibitor, and food and drink |
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| JP2013023487A (en) * | 2011-07-25 | 2013-02-04 | Maruzen Pharmaceut Co Ltd | Anti-glycation agent |
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| JP2020180070A (en) * | 2019-04-25 | 2020-11-05 | サンスター株式会社 | Anti-skin aging composition for external use |
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH06128167A (en) * | 1991-07-16 | 1994-05-10 | Shintoku Rin | Pharmaceutical composition |
| JPH0761918A (en) * | 1993-08-23 | 1995-03-07 | Tsuneo Nanba | Cosmetic |
| JP2001220313A (en) * | 2000-02-09 | 2001-08-14 | Ichimaru Pharcos Co Ltd | Cosmetic composition containing steam distillate of plant |
| JP2005082546A (en) * | 2003-09-10 | 2005-03-31 | Kanebo Cosmetics Inc | alpha-GLUCOSIDASE INHIBITOR |
| JP2006306804A (en) * | 2005-04-28 | 2006-11-09 | Kanebo Cosmetics Inc | Wrinkle formation inhibitor |
-
2007
- 2007-09-14 JP JP2007239859A patent/JP5230152B2/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH06128167A (en) * | 1991-07-16 | 1994-05-10 | Shintoku Rin | Pharmaceutical composition |
| JPH0761918A (en) * | 1993-08-23 | 1995-03-07 | Tsuneo Nanba | Cosmetic |
| JP2001220313A (en) * | 2000-02-09 | 2001-08-14 | Ichimaru Pharcos Co Ltd | Cosmetic composition containing steam distillate of plant |
| JP2005082546A (en) * | 2003-09-10 | 2005-03-31 | Kanebo Cosmetics Inc | alpha-GLUCOSIDASE INHIBITOR |
| JP2006306804A (en) * | 2005-04-28 | 2006-11-09 | Kanebo Cosmetics Inc | Wrinkle formation inhibitor |
Non-Patent Citations (2)
| Title |
|---|
| JPN6012049652; Ｉｎｏｕｙｅ， Ｈｉｒｏｙｕｋｉ ｅｔ ａｌ: 'Ｔｗｏ ｎｅｗ ｉｒｉｄｏｉｄ ｇｌｕｃｏｓｉｄｅｓ ｆｒｏｍ Ｏｓｍａｎｔｈｕｓ ｆｒａｇｒａｎｓ' Ｐｈｙｔｏｃｈｅｍｉｓｔｒｙ １４（９）, 1975, ｐｐ．２０２９-２０３２ * |
| JPN6012049653; ＰＡＰＯＵＴＳＩ Ｚｏｉ ｅｔ ａｌ: 'Ａｃｔｅｏｓｉｄｅ ａｎｄ ｍａｒｔｙｎｏｓｉｄｅ ｅｘｈｉｂｉｔ ｅｓｔｒｏｇｅｎｉｃ／ａｎｔｉｅｓｔｒｏｇｅｎｉｃ ｐｒｏｐｅｒｔｉｅｓ' Ｊ Ｓｔｅｒｏｉｄ Ｂｉｏｃｈｅｍ Ｍｏｌ Ｂｉｏｌ Ｖｏｌ．９８，Ｎｏ．１, 2006, Ｐａｇｅ．６３-７１ * |
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| CN103767975A (en) * | 2014-01-29 | 2014-05-07 | 浙江大学 | Application of osmanthus phenylethanoid glycoside extract in preparing whitening cosmetics |
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