JP2005075743A - Volatile fraction from soybean seed and composition - Google Patents
Volatile fraction from soybean seed and composition Download PDFInfo
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- JP2005075743A JP2005075743A JP2003305553A JP2003305553A JP2005075743A JP 2005075743 A JP2005075743 A JP 2005075743A JP 2003305553 A JP2003305553 A JP 2003305553A JP 2003305553 A JP2003305553 A JP 2003305553A JP 2005075743 A JP2005075743 A JP 2005075743A
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Images
Abstract
Description
本発明は、主として、皮膚傷害の治療において、皮膚疾患の治療において、細胞再生の促進において、及び体毛の増殖促進において効果を有するダイズ［Ｇｌｙｃｉｎｅ ｍａｘ（Ｌ）Ｍｅｒｒ．］の種子からの揮発性画分に関する。 The present invention mainly relates to soybeans [Glycine max (L) Merr., Which is effective in the treatment of skin injury, in the treatment of skin diseases, in the promotion of cell regeneration, and in the promotion of hair growth. ] Volatile fraction from seeds.
大豆及び黒大豆を包含する、ダイズ［Ｇｌｙｃｉｎｅ ｍａｘ（Ｌ）Ｍｅｒｒｉｌｌ］は、世界の油及びタンパク質の最も重要な供給源である。例えば、大豆は、加工処理して、サラダオイルとして、又はマーガリン及びショートニングの製造用として使用される食用の半乾性油を得ることができる。大豆油は、又、塗料、リノリウム、オイルクロス、印刷インキ、石鹸、殺虫剤及び消毒薬の製造において使用することができる。なお又、油工業の副産物から得られるレシチンリン脂質は、食品、化粧品、医薬、皮革、塗料、プラスチックス、石鹸及び洗剤において、湿潤剤及び安定化剤として使用することができる。大豆粕は、非常にタンパク質の豊富な家畜用の飼料である。更に、大豆タンパク質は、合成繊維、接着剤、織物の糊づけ、防水加工、消火用泡、その他の製造において使用することができる。 Soybean [Glycine max (L) Merrill], including soy and black soybeans, is the most important source of oil and protein in the world. For example, soybeans can be processed to obtain an edible semi-drying oil that is used as a salad oil or for the production of margarine and shortening. Soybean oil can also be used in the manufacture of paints, linoleum, oil cloth, printing inks, soaps, insecticides and disinfectants. In addition, lecithin phospholipids obtained from by-products of the oil industry can be used as wetting agents and stabilizers in foods, cosmetics, medicines, leather, paints, plastics, soaps and detergents. Soybean meal is a very protein-rich livestock feed. Furthermore, soy protein can be used in the production of synthetic fibers, adhesives, textile gluing, waterproofing, fire-fighting foams, and others.
医学的使用においては、大豆は多くの疾病に効果を有することが報告されている。 In medical use, soy has been reported to be effective against many diseases.
大豆は、腸、心臓、腎臓、肝臓及び胃の機能を調節するための栄養補助食品として使用することができる。大豆油は不飽和脂肪酸を多く含んでいるので、高コレステロール血症と闘うために使用することができる。大豆からの医薬レシチンは、抗脂肝剤として機能する。更に、抗硬直因子として知られるスチグマステロール及びある種の抗高血圧症薬におけるジオスゲニンの代わりとして使用されるシトステロールは大豆から調製された。大豆において発見されたイソフラボン及び植物エストロゲンは、乳癌、前立腺癌及び大腸癌を含む種々の癌に対して予防効果を有することが示唆された（非特許文献１）。植物ステロール添加マーガリンの消費は、又、中高年の高コレステロール血症者の総血漿コレステロール及びＬＤＬ−コレステロール濃度を低減することが見出された（非特許文献２）。 Soy can be used as a dietary supplement to regulate intestinal, heart, kidney, liver and stomach function. Soybean oil is high in unsaturated fatty acids and can be used to combat hypercholesterolemia. Pharmaceutical lecithin from soybeans functions as an anti-fatty liver agent. In addition, stigmasterol, known as an anti-rigor factor, and sitosterol, used as a replacement for diosgenin in certain antihypertensive drugs, were prepared from soybeans. It has been suggested that isoflavones and phytoestrogens found in soybean have a preventive effect against various cancers including breast cancer, prostate cancer and colon cancer (Non-patent Document 1). Consumption of plant sterol-added margarine has also been found to reduce total plasma cholesterol and LDL-cholesterol levels in middle-aged and hypercholesterolemic individuals (Non-Patent Document 2).
大豆からのある種の抽出物は、又、医薬効果を有することが報告されている。茶大豆種、秋田在来の種皮から得た７０％水性アセトン抽出物の、１，１−ジフェニル−２−ピクリルヒドラジル（ＤＰＰＨ）ラジカルに対する捕捉活性が開示されている（非特許文献３）。６０℃未満で粉末鉱石の下でゆっくりと焙煎し、Ａｓｐｅｒｇｉｌｌｕｓ ｏｒｙｚａｅで三日間にわたって発酵して各成分を低分子物質に変換した胚芽抽出物、大豆、米糠、ティア グラス（ｔｅａｒ ｇｒａｓｓ）、ゴマ、小麦、シトロン、緑茶、緑葉抽出物及び発芽米からの抽出物は、抗酸化効果を有することが見出された（非特許文献４）。ニンジン、アブラナの緑葉、及び黒豆、アズキ、緑豆、及び大豆を含む豆類の煮沸抽出物は、高い抗ガン促進及びラジカル捕捉活性を有することが示された（非特許文献５）。黒豆の水抽出物も、又、ラットにおける血清グルタミン酸−オキサロ酢酸トランスアミナーゼ（ｓＧＯＴ）及び血清グルタミン酸−ピルビン酸トランスアミナーゼ（ｓＧＰＴ）活性を測定することによって、アセトアミノフェン誘発肝傷害に対する効果が報告されている（非特許文献６）。低級脂肪族アルコール及び低沸点エステルの混合系からなる抽出液での大豆種子の抽出物が、渇きを癒すためのリフレッシュドリンク剤として示された（特許文献１）。 Certain extracts from soy have also been reported to have medicinal effects. A scavenging activity for 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals of a 70% aqueous acetone extract obtained from seed coats of tea soybean seeds and Akita natives is disclosed (Non-patent Document 3). . Germ extract, soybean, rice bran, tear glass, sesame seeds, roasted slowly under powdered ore below 60 ° C, fermented with Aspergillus oryzae for 3 days to convert each component to low molecular weight material It has been found that extracts from wheat, citron, green tea, green leaf extract and germinated rice have an antioxidant effect (Non-Patent Document 4). It has been shown that boiled extracts of carrots, rape green leaves, and beans including black beans, azuki beans, mung beans, and soybeans have high anti-cancer promoting and radical scavenging activities (Non-Patent Document 5). Black bean water extract has also been reported to have effects on acetaminophen-induced liver injury by measuring serum glutamate-oxaloacetate transaminase (sGOT) and serum glutamate-pyruvate transaminase (sGPT) activities in rats. (Non-patent document 6). An extract of soybean seeds in an extract composed of a mixed system of a lower aliphatic alcohol and a low-boiling ester has been shown as a refreshing drink for healing thirst (Patent Document 1).
大豆からのある種の特別な抽出物が、ある種の皮膚疾患の治療における化粧品又は医薬に適用されることが見出された。 It has been found that certain special extracts from soy apply to cosmetics or medicines in the treatment of certain skin diseases.
一定の比率でスフィンゴミエリン及びリン脂質を含有する大豆抽出物が、乾燥肌の治療用の化粧品に使用されることが開示された（特許文献２）。そのような抽出物は、熟した全粒大豆又は脱脂大豆粉を、脂肪族アルコールのみを使用して又は水との混合物で抽出し、次いで脂肪族炭化水素及び脂肪族ケトンで処理することによって製造された。それ故、抽出物は脂質可溶性である。 It has been disclosed that soybean extract containing sphingomyelin and phospholipid at a certain ratio is used in cosmetics for treating dry skin (Patent Document 2). Such extracts are produced by extracting ripe whole soybeans or defatted soybean flour using only aliphatic alcohols or in a mixture with water and then treating with aliphatic hydrocarbons and aliphatic ketones. It was done. Therefore, the extract is lipid soluble.
乳清（ホエー）及びキハダ抽出物から選択される一つ又はそれ以上の植物抽出物、及び更にコガネバナ、コンフリー及びダイズから選択される一つ又はそれ以上の抽出物を含有するざ瘡薬、面皰産生阻害剤又は化粧料組成物が、ざ瘡又はざ瘡に起因する炎症性のひび割れた皮膚のような皮膚疾患の予防又は治療に効果があることが見出された（特許文献３）。組成物は、一般の飲料、食品、その他の製造に使用することができる。 Acne medicines containing one or more plant extracts selected from whey and yellowfin extracts, and also one or more extracts selected from Scarab, Comfrey and soybeans, It has been found that a comedone production inhibitor or a cosmetic composition is effective in preventing or treating skin diseases such as acne or inflammatory cracked skin caused by acne (Patent Document 3). The composition can be used for production of general beverages, foods, and the like.
大豆又は大豆の水抽出液から得られたマロニルイソフラボングリコシド及び、例えば、マロニルダイジン等は、上皮細胞増殖促進剤として使用された（特許文献４）。グリコシドは、４５〜６５℃で２〜４時間、苛性ソーダでｐＨ７．５〜ｐＨ９．０に調節した水の中に皮をむいた大豆を液浸し、大豆を除去して水抽出溶液として浸漬水を得、限外ろ過膜によって抽出溶液からタンパク質を除去してろ液を得、ろ液を吸着剤と接触させ、そしてアルコールの水性溶液又はアルコールのアルカリ性水性溶液を使用することによって吸着した物質からグリコシドを溶出することによって得ることができる。 Malonyl isoflavone glycoside obtained from soybean or soybean aqueous extract and, for example, malonyl daidzine and the like were used as epithelial cell growth promoters (Patent Document 4). Glycoside is soaked in peeled soybeans in water adjusted to pH 7.5 to pH 9.0 with caustic soda at 45 to 65 ° C. for 2 to 4 hours. Obtain the filtrate from the extraction solution by ultrafiltration membrane to obtain a filtrate, contact the filtrate with an adsorbent, and remove glycosides from the adsorbed material by using an aqueous solution of alcohol or an alkaline aqueous solution of alcohol. It can be obtained by elution.
微生物による発酵大豆製品は、又、抗活性酸素作用組成物、薬剤、食品、化粧品及び医薬品として適用されている（例えば、特許文献５）。 Fermented soy products by microorganisms are also applied as anti-active oxygen action compositions, drugs, foods, cosmetics and pharmaceuticals (for example, Patent Document 5).
しかしながら、いずれの公知技術も、ダイズからの低濃度のアルコールを含む或いは含まない揮発性画分が、皮膚傷害及び皮膚疾患を治療するために容易に調製されることを教示又は示唆していない。
本発明は、皮膚傷害の治療、皮膚疾患の治療、細胞の再生増進及び体毛の増殖促進の劇的な効果を示す、ダイズの種子からの揮発性画分を提供するものである。 The present invention provides a volatile fraction from soybean seeds that exhibits dramatic effects of treating skin injury, treating skin diseases, enhancing cell regeneration and promoting hair growth.
本発明の揮発性画分は、
（ａ）約１５重量％未満の濃度のアルコールを含有するアルコール溶液中、又は水中に種子の粗抽出物を供給し；そして
（ｂ）約１気圧未満の気圧及び約１１０℃未満の温度で粗抽出物を蒸発させて、揮発性画分を得る；
ステップを含むプロセスによって調製される。
The volatile fraction of the present invention is
(A) supplying a crude extract of seeds in an alcohol solution containing alcohol at a concentration of less than about 15% by weight or in water; and (b) crude at a pressure of less than about 1 atmosphere and a temperature of less than about 110 ° C. Evaporate the extract to obtain a volatile fraction;
Prepared by a process comprising steps.
該揮発性画分は、いずれの公知のダイズの種子抽出物とも異なっている。何故なら、高速液体クロマトグラフィー（ＨＰＬＣ）のパターンは独特のプロフィールを示しているからである。特に、内径（Ｉ．Ｄ．）４．６ｍｍ、長さ（Ｌ）２５０ｍｍ及び粒子径５μｍのＷａｔｅｒｓ Ｓｐｈｅｒｓｏｒｂ（ＴＭ）−ＯＤＳ２カラムを用いて２００ｎｍで測定した、揮発性画分５０μｌのＨＰＬＣスペクトルは、変動係数８％で、保持時間２．９１０、５．１９０、１３．１９０、及び５０．８１５分にピークを有する。ただし、この場合の移動相は、０から５分が９５％Ｈ2Ｏ／５％ＣＨ3ＣＮ；５分から３０分が９５％Ｈ2Ｏ／５％ＣＨ3ＣＮの勾配；３０分から８０分が７５％Ｈ2Ｏ／２５％ＣＨ3ＣＮの勾配；８０分から１００分が０％Ｈ2Ｏ／１００％ＣＨ3ＣＮの勾配；１００分から１０１分が０％Ｈ2Ｏ／１００％ＣＨ3ＣＮの勾配；及び１０１分が９５％Ｈ2Ｏ／５％ＣＨ3ＣＮであり、流速は１ｍＬ／分である。 The volatile fraction is different from any known soybean seed extract. This is because the pattern of high performance liquid chromatography (HPLC) shows a unique profile. In particular, the HPLC spectrum of a 50 μl volatile fraction, measured at 200 nm using a Waters Sphersorb (TM) -ODS2 column with an inner diameter (ID) of 4.6 mm, a length (L) of 250 mm and a particle size of 5 μm, It has a coefficient of variation of 8% and peaks at retention times of 2.910, 5.190, 13.190, and 50.815 minutes. However, mobile phase in this case is 5 minutes from 0 95% H 2 O / 5% CH 3 CN; from 30 minutes to 80 minutes; 5 to 30 minutes is 95% H 2 O / 5% CH 3 CN gradient of 75% H 2 O / 25% CH 3 CN gradient; 80 to 100 minutes from 0% H 2 O / 100% CH 3 CN gradient; 100 to 101 minutes from 0% H 2 O / 100% CH 3 CN Gradient; and 101 minutes is 95% H 2 O / 5% CH 3 CN and the flow rate is 1 mL / min.
本発明は、又、本発明の揮発性画分を含む組成物を提供する。特に、該組成物は、医薬組成物、化粧品組成物又は皮膚の洗浄用組成物に使用することができる。 The present invention also provides a composition comprising the volatile fraction of the present invention. In particular, the composition can be used in pharmaceutical compositions, cosmetic compositions or skin cleansing compositions.
本発明によれば、ダイズ［Ｇｌｙｃｉｎｅ ｍａｘ（Ｌ）Ｍｅｒｒ．］の揮発性画分は、皮膚傷害の治療、皮膚障害の治療、細胞の再生増進、及び体毛の増殖促進において劇的な効果を有することが、意外にも見出された。 According to the present invention, soybean [Glycine max (L) Merr. It has been surprisingly found that the volatile fraction of] has dramatic effects in treating skin injury, treating skin disorders, enhancing cell regeneration, and promoting hair growth.
本発明によれば、ダイズの種子からの揮発性画分は、
（ａ）約１５重量％未満の濃度のアルコールを含有するアルコール溶液中、又は水中に種子の粗抽出物を供給し；そして
（ｂ）約１気圧未満の気圧及び約１１０℃未満の温度で粗抽出物を蒸発させて、揮発性画分を得る；
ステップを含むプロセスによって調製される。
According to the present invention, the volatile fraction from soybean seeds is:
(A) supplying a crude extract of seeds in an alcohol solution containing alcohol at a concentration of less than about 15% by weight or in water; and (b) crude at a pressure of less than about 1 atmosphere and a temperature of less than about 110 ° C. Evaporate the extract to obtain a volatile fraction;
Prepared by a process comprising steps.
アルコールを約１５％未満の濃度で含有するアルコール溶液中又は水中の種子の粗抽出物から得られる揮発性画分が、皮膚科学における効果を有することが意外にも発見された。該揮発性画分は容易に調製される。ダイズの種子から調製された医薬又は化粧品の製品は、（１）大豆が長年消費されてきたこと、そして非毒性であると信じられること、及び（２）大豆は高価でないことから有利である。 It has been surprisingly discovered that volatile fractions obtained from crude extracts of seeds in alcoholic solutions or water containing alcohol at a concentration of less than about 15% have an effect in dermatology. The volatile fraction is easily prepared. Pharmaceutical or cosmetic products prepared from soy seeds are advantageous because (1) soy has been consumed for many years and is believed to be non-toxic, and (2) soy is not expensive.
本発明の実施態様において、ダイズの種子からの粗抽出物は、約１５重量％未満の濃度のアルコール溶液又は水を用いた抽出によって得られる。当業者により通常使用される植物種子から粗抽出物を得るいかなる方法も、本発明の実施に使用することができる。例えば、粗抽出物は、種子を、粉砕、撹拌、攪乱、切断又は切り刻みのような、いかなる通常の手段であれ小片に細分化し、約１５重量％未満の濃度のアルコール溶液に浸漬することによって得ることができる。好ましい実施態様において、アルコールの濃度は、約１０重量％未満、より好ましくは約３重量％又は約２重量％である。本発明によれば、粗抽出物は、又、水で抽出することもできる。好ましい実施態様において、種子片は、約５重量部から約１０重量部の範囲のアルコール又は水に浸漬される。当業者によって通常使用される、種子を細分化し細分化した種子から抽出するいかなる方法も、本発明の実施に使用することができる。 In an embodiment of the present invention, the crude extract from soybean seeds is obtained by extraction with an alcohol solution or water having a concentration of less than about 15% by weight. Any method of obtaining a crude extract from plant seeds commonly used by those skilled in the art can be used in the practice of the present invention. For example, the crude extract is obtained by subdividing the seed into small pieces by any conventional means, such as grinding, stirring, perturbing, cutting or chopping, and soaking in an alcohol solution at a concentration of less than about 15% by weight. be able to. In preferred embodiments, the concentration of alcohol is less than about 10% by weight, more preferably about 3% by weight or about 2% by weight. According to the invention, the crude extract can also be extracted with water. In a preferred embodiment, the seed pieces are immersed in a range of about 5 to about 10 parts by weight alcohol or water. Any method of subdividing seeds and extracting from the subdivided seeds commonly used by those skilled in the art can be used in the practice of the present invention.
本発明によれば、粗抽出物は、約１気圧未満の気圧及び約１１０℃未満の温度、好ましくは約６０℃から約１１０℃で、蒸発される。揮発性画分は、蒸気を冷却することによって液体の形で採取することができる。 According to the present invention, the crude extract is evaporated at a pressure of less than about 1 atmosphere and a temperature of less than about 110 ° C., preferably from about 60 ° C. to about 110 ° C. The volatile fraction can be collected in liquid form by cooling the vapor.
本発明の好ましい実施態様において、一定の気圧及び温度で粗抽出物を蒸発し、その蒸気を冷却することによって該揮発性画分を採取するプロセスは、ロータリーエバポレーターで実施することができ、ここで、蒸気は、冷水が供給される凝縮管に蒸発され、そして蒸気は凝縮管を通過して冷却され、揮発性画分が液状で採取される。操作は単純でコストは低い。 In a preferred embodiment of the invention, the process of collecting the volatile fraction by evaporating the crude extract at a constant pressure and temperature and cooling the vapor can be carried out on a rotary evaporator, where The vapor is evaporated into a condenser tube fed with cold water, and the vapor is cooled through the condenser tube and the volatile fraction is collected in liquid form. Operation is simple and low cost.
本発明による揮発性画分は、高速液体クロマトグラフィー（ＨＰＬＣ）で分析され、同定することができる。例えば、内径（Ｉ．Ｄ．）４．６ｍｍ、長さ（Ｌ）２５０ｍｍ及び粒子径５μｍのＷａｔｅｒｓ Ｓｐｈｅｒｓｏｒｂ（ＴＭ）−ＯＤＳ２カラムを用いて２００ｎｍで測定した、揮発性画分５０μｌのＨＰＬＣスペクトルは、変動係数８％で、保持時間２．９１０、５．１９０、１３．１９０、及び５０．８１５分にピークを有する。ただし、この場合の移動相は、０から５分が９５％Ｈ2Ｏ／５％ＣＨ3ＣＮ；５分から３０分が９５％Ｈ2Ｏ／５％ＣＨ3ＣＮの勾配；３０分から８０分が７５％Ｈ2Ｏ／２５％ＣＨ3ＣＮの勾配；８０分から１００分が０％Ｈ2Ｏ／１００％ＣＨ3ＣＮの勾配；１００分から１０１分が０％Ｈ2Ｏ／１００％ＣＨ3ＣＮの勾配；及び１０１分が９５％Ｈ2Ｏ／５％ＣＨ3ＣＮであり、流速は１ｍＬ／分である。スペクトルによれば、揮発性画分中の物質は強い極性を示す。それらは、公知文献に開示された他の成分とは大きく異なっている。例えば、ダイゼイン（ｄａｉｄｚｅｉｎ）及びゲニステイン（ｇｅｎｉｓｔｅｉｎ）は、その構造中に四つの環を有し、弱い極性を有する。 The volatile fraction according to the invention can be analyzed and identified by high performance liquid chromatography (HPLC). For example, the HPLC spectrum of a 50 μl volatile fraction measured at 200 nm using a Waters Sphersorb (TM) -ODS2 column with an inner diameter (ID) of 4.6 mm, a length (L) of 250 mm and a particle size of 5 μm is It has a coefficient of variation of 8% and peaks at retention times of 2.910, 5.190, 13.190, and 50.815 minutes. However, mobile phase in this case is 5 minutes from 0 95% H 2 O / 5% CH 3 CN; from 30 minutes to 80 minutes; 5 to 30 minutes is 95% H 2 O / 5% CH 3 CN gradient of 75% H 2 O / 25% CH 3 CN gradient; 80 to 100 minutes from 0% H 2 O / 100% CH 3 CN gradient; 100 to 101 minutes from 0% H 2 O / 100% CH 3 CN Gradient; and 101 minutes is 95% H 2 O / 5% CH 3 CN and the flow rate is 1 mL / min. According to the spectrum, the substances in the volatile fraction are strongly polar. They are very different from other components disclosed in the known literature. For example, daidzein and genistein have four rings in their structure and are weakly polar.
本発明の一つの好ましい実施態様において、皮膚傷害の治療、皮膚疾患の治療、細胞の再生増進及び体毛の増殖促進において、ダイズの種子を補助するために幾種類かの薬草が加えられる。本発明によれば、揮発性画分は、ダイズの種子、及び、ゴマの種子、リョクトウの種子及び抗菌薬草から成る群から選択される少なくとも一つの材料を含む原料から得られる。該原料からの揮発性画分の調製並びに揮発性画分の分析及び同定は、上記に記載されている。好ましくは、抗菌薬草は、掌葉大黄の乾燥根茎、ダイオウの乾燥根茎、キハダの乾燥樹皮及びオウバクの乾燥樹皮からなる群から選択される。別の態様において、原料は、好ましくは、約４０から約１００％のダイズの種子、０から約５０％のゴマの種子、０から約５０％のリョクトウの種子及び０から約３０％の抗菌薬草を含む。 In one preferred embodiment of the invention, several types of herbs are added to assist soybean seeds in treating skin injury, treating skin diseases, promoting cell regeneration and promoting hair growth. According to the invention, the volatile fraction is obtained from a raw material comprising soybean seeds and at least one material selected from the group consisting of sesame seeds, mungbean seeds and antibacterial herbs. Preparation of the volatile fraction from the raw material and analysis and identification of the volatile fraction are described above. Preferably, the antibacterial medicinal herb is selected from the group consisting of dried rhizomes of palm leaves, yellow rhizomes, dried bark of yellowfin and dried bark of buckwheat. In another embodiment, the ingredient is preferably about 40 to about 100% soybean seed, 0 to about 50% sesame seed, 0 to about 50% mungbean seed and 0 to about 30% antimicrobial herb. including.
本発明の更に好ましい実施態様の一つにおいて、原料は、約７０％のダイズの種子、約２０％のゴマの種子、約５％の掌葉大黄又はダイオウの乾燥根茎、及び約５％のキハダ又はオウバクの乾燥樹皮を含む。該原料は約２％のアルコール中に浸漬される。 In one more preferred embodiment of the present invention, the feedstock is about 70% soybean seed, about 20% sesame seed, about 5% palm foliage or dairy dry rhizome, and about 5% yellowfin. Or it contains dried bark of Aoba. The raw material is immersed in about 2% alcohol.
本発明の別の好ましい実施態様において、原料は、約８０％のダイズの種子及び約２０％のリョクトウの種子を含む。該原料は約２％のアルコール中に浸漬される。 In another preferred embodiment of the invention, the feedstock comprises about 80% soybean seed and about 20% mungbean seed. The raw material is immersed in about 2% alcohol.
本発明の更に別の好ましい実施態様において、原料は、約８０％のダイズの種子及び約２０％のゴマの種子を含む。該原料は約２％のアルコール中に浸漬される。 In yet another preferred embodiment of the invention, the raw material comprises about 80% soybean seed and about 20% sesame seed. The raw material is immersed in about 2% alcohol.
本発明の更に別の好ましい実施態様において、原料は、約９０％のダイズの種子及び約１０％の掌葉大黄又はダイオウの乾燥根茎を含む。該原料は水中に浸漬される。 In yet another preferred embodiment of the present invention, the feedstock comprises about 90% soybean seed and about 10% palm leaf yellow or dairy dried rhizome. The raw material is immersed in water.
本発明の更に別の好ましい実施態様において、原料は、約８０％のダイズの種子、約１０％のゴマの種子及び約１０％の掌葉大黄又はダイオウの乾燥根茎を含む。該原料は水中に浸漬される。 In yet another preferred embodiment of the present invention, the feedstock comprises about 80% soybean seed, about 10% sesame seed and about 10% palm foliage or dairy dried rhizome. The raw material is immersed in water.
本発明の最も好ましい実施態様において、原料は、約５３％のダイズの種子、約２０％のゴマの種子、約１７％のリョクトウの種子、約５％の掌葉大黄又はダイオウの乾燥根茎及び約５％のキハダ又はオウバクの乾燥樹皮を含む。該原料は約３％又は約２％のアルコール中又は水中に浸漬される。更に、内径（Ｉ．Ｄ．）４．６ｍｍ、長さ（Ｌ）２５０ｍｍ及び粒子径５μｍのＷａｔｅｒｓ Ｓｐｈｅｒｓｏｒｂ（ＴＭ）−ＯＤＳ２カラムを用いて２００ｎｍで測定した、揮発性画分５０μｌのＨＰＬＣスペクトルは、変動係数８％で、保持時間２．８９６、５．１１１、１２．９４４、及び４９．６０３分にピークを有する。ただし、この場合の移動相は、０から５分が９５％Ｈ2Ｏ／５％ＣＨ3ＣＮ；５分から３０分が９５％Ｈ2Ｏ／５％ＣＨ3ＣＮの勾配；３０分から８０分が７５％Ｈ2Ｏ／２５％ＣＨ3ＣＮの勾配；８０分から１００分が０％Ｈ2Ｏ／１００％ＣＨ3ＣＮの勾配；１００分から１０１分が０％Ｈ2Ｏ／１００％ＣＨ3ＣＮの勾配；及び１０１分が９５％Ｈ2Ｏ／５％ＣＨ3ＣＮであり、流速は１ｍＬ／分である。 In the most preferred embodiment of the present invention, the feedstock is about 53% soybean seed, about 20% sesame seed, about 17% mungbean seed, about 5% palm foliage or dairy dry rhizome and about Contains 5% yellowfin or agar dry bark. The raw material is immersed in about 3% or about 2% alcohol or water. Furthermore, the HPLC spectrum of 50 μl of volatile fraction, measured at 200 nm using a Waters Sphersorb (TM) -ODS2 column with an inner diameter (ID) of 4.6 mm, a length (L) of 250 mm and a particle diameter of 5 μm, It has a coefficient of variation of 8% and peaks at retention times of 2.896, 5.111, 12.944, and 49.603 minutes. However, mobile phase in this case is 5 minutes from 0 95% H 2 O / 5% CH 3 CN; from 30 minutes to 80 minutes; 5 to 30 minutes is 95% H 2 O / 5% CH 3 CN gradient of 75% H 2 O / 25% CH 3 CN gradient; 80 to 100 minutes from 0% H 2 O / 100% CH 3 CN gradient; 100 to 101 minutes from 0% H 2 O / 100% CH 3 CN Gradient; and 101 minutes is 95% H 2 O / 5% CH 3 CN and the flow rate is 1 mL / min.
本発明は、又、本発明による揮発性画分を含む組成物を提供する。好ましくは、組成物は、医薬組成物、化粧品組成物及び皮膚の洗浄用組成物から成る群から選択される。本発明による好ましい実施態様において、組成物は、更に、ボルネオール等の抗菌剤を含む。好ましくは、本発明による揮発性画分の量は、全組成物重量の約７０％から約８０％の範囲である。 The present invention also provides a composition comprising a volatile fraction according to the present invention. Preferably, the composition is selected from the group consisting of a pharmaceutical composition, a cosmetic composition and a skin cleansing composition. In a preferred embodiment according to the present invention, the composition further comprises an antibacterial agent such as borneol. Preferably, the amount of volatile fraction according to the present invention ranges from about 70% to about 80% of the total composition weight.
本発明の一つの実施態様において、組成物は、人に容易に適用される軟膏を形成するボルネオール、ステアリン酸、ステアリルアルコール、パルミチン酸、セチルアルコール、蜜ろう及びポリオキシエチレンソルビタンモノステアリン酸エステル（ＰＯＥ）から選ばれる一種又は二種以上を更に含む。 In one embodiment of the present invention, the composition comprises borneol, stearic acid, stearyl alcohol, palmitic acid, cetyl alcohol, beeswax and polyoxyethylene sorbitan monostearate (form) that forms an ointment that is easily applied to humans. 1 type or 2 types or more chosen from POE) are further included.
皮膚傷害又は皮膚疾患を治療する方法は、皮膚傷害の治療、皮膚疾患の治療、細胞の再生増進、及び体毛の増殖を促進するために有効な十分な量で、本発明による組成物を適用することを含む。 A method of treating skin injury or skin disease applies a composition according to the invention in a sufficient amount effective to treat skin injury, treatment of skin disease, promote cell regeneration, and hair growth. Including that.
本発明に記載の揮発性画分は、皮膚傷害の治療、皮膚疾患の治療、細胞の再生増進、及び体毛の増殖促進に有用である。例えば、皮膚傷害又は皮膚疾患は、熱傷害、日焼け、照射性傷害、急性及び慢性外傷、斑状出血、皮膚炎、斑、丘疹、小結節、小水疱、気腫性嚢胞、膿疱、膨疹、斑点、嚢胞、鱗屑、痂皮、ひび、潰瘍、ざ瘡、乾皮症、落屑、そう痒、アレルギー性皮膚炎、皮疹及び毛嚢欠損を包含する。好ましくは、本発明による組成物は、熱傷害、照射性傷害、急性及び慢性外傷、皮膚炎、壊疽、慢性うっ血性潰瘍、擦過傷、アトピー性皮膚炎、縫合創傷、糖尿病創傷及び毛嚢欠損を治療するのに効果的である。 The volatile fraction described in the present invention is useful for treating skin injury, treating skin diseases, enhancing cell regeneration, and promoting hair growth. For example, skin injury or skin disease may include heat injury, sunburn, radiation injury, acute and chronic trauma, ecchymosis, dermatitis, plaques, papules, nodules, blisters, emphysematous cysts, pustules, rashes, spots, Includes cysts, scales, crusts, cracks, ulcers, acne, psoriasis, desquamation, pruritus, allergic dermatitis, rashes and hair follicle defects. Preferably, the composition according to the invention treats heat injury, radiation injury, acute and chronic trauma, dermatitis, gangrene, chronic congestive ulcer, abrasion, atopic dermatitis, suture wound, diabetic wound and hair follicle defect It is effective to do.
以下に記載する動物モデルにおいて、本発明による揮発性画分は、肉芽、傷痕形成性及びアレルギーなしに大領域の皮膚欠損を治癒するのに効果的であることが証明された。更に、毛嚢欠損は治癒することができ、体毛は紅斑又は焼痂なしに再び増殖することができた。本発明による揮発性画分で治療された創傷の治癒率及び状態は、従来の医薬よりもよりよいものである。本発明による揮発性画分で治療された創傷は、瘢痕なしに治癒し、治癒した創傷の皮膚の色は、正常な皮膚の色と同じであった。更に、組織学的試験において、本発明による揮発性画分で治療された創傷の皮膚組織は、通常、外観に大きな障害と複雑な再建手術を生ずる原因となる線維性変性を示さなかった。 In the animal model described below, the volatile fraction according to the present invention has proven effective in healing large areas of skin defects without granulation, scar formation and allergies. In addition, hair follicle defects could be healed and hair could grow again without erythema or cauterization. The healing rate and condition of wounds treated with the volatile fraction according to the invention are better than conventional medicines. The wound treated with the volatile fraction according to the invention healed without scarring and the skin color of the healed wound was the same as the normal skin color. Furthermore, in histological examination, the skin tissue of wounds treated with the volatile fraction according to the present invention usually did not show fibrotic degeneration that caused a significant visual impairment and complex reconstructive surgery.
更に、アレルギー反応、炎症、膿疱又は紅疹は、本発明による揮発性画分で治療された創傷には見出されなかった。治療後、創傷の副器官及び細胞は、正常の皮膚と同じに増殖した。対照的に、従来の医薬で治療した創傷は、副器官及び細胞の数はより少なく、創傷の線維幅は正常よりも大きかった。 Furthermore, no allergic reaction, inflammation, pustules or erythema were found in wounds treated with the volatile fraction according to the invention. After treatment, the wound's accessory organs and cells grew the same as normal skin. In contrast, wounds treated with conventional medication had fewer accessory organs and cells, and wound fiber width was greater than normal.
更に、本発明による揮発性画分は、照射傷害の治療にも効果的である。これまで、照射傷害の治療に有効な薬物はなかった。照射傷害にかかった人において、揮発性画分で治療された後の創傷は、体毛にほとんど損傷を示さず、アレルギーは示さないことが、意外にも見出された。 Furthermore, the volatile fraction according to the invention is also effective in the treatment of radiation injury. To date, no drug has been effective in treating radiation injury. It was surprisingly found that in persons suffering from radiation injury, the wound after treatment with the volatile fraction showed little damage to the hair and no allergies.
更に、創傷治癒において通常見られる炎症も組織の萎縮も、本発明による揮発性画分で治療された創傷には見出されなかった。揮発性画分は、皮膚が損傷された創傷の治癒に効果的である。それ故、本発明は、大きな領域と皮膚創傷を治療するのに、よりよい、便利な、そして経済的な方法を提供するものである。 Furthermore, neither inflammation nor tissue atrophy normally seen in wound healing was found in wounds treated with the volatile fraction according to the invention. The volatile fraction is effective in healing wounds with damaged skin. Therefore, the present invention provides a better, convenient and economical method for treating large areas and skin wounds.
以下の実施例は、例示のみの目的のものであり、本発明の範囲を限定することを意図するものではない。 The following examples are for illustrative purposes only and are not intended to limit the scope of the present invention.
実施例１
ダイズ［Ｇｌｙｃｉｎｅ ｍａｘ（Ｌ）Ｍｅｒｒ．］の種子からの揮発性画分の調製
ダイズ［Ｇｌｙｃｉｎｅ ｍａｘ（Ｌ）Ｍｅｒｒ．］の種子、ゴマ（Ｓｅｓａｍｕｍ ｉｎｄｉｃｕｍ）の種子、リョクトウ（Ｐｈａｓｅｏｌｕｓ ｒａｄｉａｔｕｓ Ｌ．）の種子、掌葉大黄（Ｒｈｅｕｍ ｐａｌｍａｔｕｍ）又はダイオウ（大黄、Ｒｈｅｉ Ｒｉｓｏｍａ）の乾燥根茎及びキハダ（Ｐｈｅｌｌｏｄｅｎｄｒｏｎ ａｍｕｒｅｎｓｅ Ｒｕｐｒ．）又はオウバク（黄柏、Ｐｈｅｌｌｏｄｅｎｄｒｉ Ｃｏｒｔｅｘ）の乾燥樹皮を含む原料を粉にし、アルコール又は蒸留水８重量部で抽出して、粗抽出物を得た。含量を表１に示す。揮発性画分は、１気圧未満の圧力、６０℃の温度で、冷水を供給した濃縮管を通過させるロータリーエバポレーター（ＥＹＥＬＡ Ｎ−１０００Ｓ、１０００Ｓ−Ｗ）を用いて、粗抽出物を蒸発させることによって得た。
Example 1
Soybean [Glycine max (L) Merr. Of volatile fraction from seeds of soybean [Glycine max (L) Merr. ], Sesame indicum seed, mungbean (Phaseolus radiatus L.) seed, palm leaf (Rheum palmatum) or dried rhizome (Rhei Rendoma) or Phellodendronum A raw material containing dried bark of (yellow twilight, Phellodendri Cortex) was powdered and extracted with 8 parts by weight of alcohol or distilled water to obtain a crude extract. The contents are shown in Table 1. The volatile fraction is obtained by evaporating the crude extract using a rotary evaporator (EYELA N-1000S, 1000S-W) passing through a concentration tube supplied with cold water at a pressure of less than 1 atm and a temperature of 60 ° C. Obtained by.
画分１の５０μｌのＨＰＬＣの結果を図１に示す。ＨＰＬＣは、内径（Ｉ．Ｄ．）４．６ｍｍ、長さ（Ｌ）２５０ｍｍ及び粒子径５μｍのＷａｔｅｒｓ Ｓｐｈｅｒｓｏｒｂ（ＴＭ）−ＯＤＳ２カラムで、移動相は、０から５分が９５％Ｈ2Ｏ／５％ＣＨ3ＣＮ；５分から３０分が９５％Ｈ2Ｏ／５％ＣＨ3ＣＮの勾配；３０分から８０分が７５％Ｈ2Ｏ／２５％ＣＨ3ＣＮの勾配；８０分から１００分が０％Ｈ2Ｏ／１００％ＣＨ3ＣＮの勾配；１００分から１０１分が０％Ｈ2Ｏ／１００％ＣＨ3ＣＮの勾配；及び１０１分が９５％Ｈ2Ｏ／５％ＣＨ3ＣＮであり、流速は１ｍＬ／分で行われた。ＨＰＬＣは、Ｗａｔｅｒｓ Ａｌｌｉａｎｃｅ２７９５ＨＴで行われ、スペクトル測定は、Ｗａｔｅｒｓ Ｐｈｏｔｏ Ｄｉｏｄｅ Ａｒｒａｙ２９９６によって１９０から４００ｎｍで行われた。２００ｎｍで測定したスペクトルに示すように、変動係数８％で、保持時間２．９１０、５．１９０、１３．１９０、及び５０．８１５分に鋭いピークがあった。
The result of 50 μl HPLC of fraction 1 is shown in FIG. The HPLC was a Waters Sphersorb (TM) -ODS2 column with an inner diameter (ID) of 4.6 mm, a length (L) of 250 mm and a particle size of 5 μm, and the mobile phase was 95% H 2 O / 0 from 0 to 5 minutes. 5% CH 3 CN; 5 to 30 minutes 95% H 2 O / 5% CH 3 CN gradient; 30 to 80 minutes 75% H 2 O / 25% CH 3 CN gradient; 80 to 100
画分３の５０μｌのＨＰＬＣの結果を図２に示す。ＨＰＬＣは、内径（Ｉ．Ｄ．）４．６ｍｍ、長さ（Ｌ）２５０ｍｍ及び粒子径５μｍのＷａｔｅｒｓ Ｓｐｈｅｒｓｏｒｂ（ＴＭ）−ＯＤＳ２カラムで、移動相は、０から５分が９５％Ｈ2Ｏ／５％ＣＨ3ＣＮ；５分から３０分が９５％Ｈ2Ｏ／５％ＣＨ3ＣＮの勾配；３０分から８０分が７５％Ｈ2Ｏ／２５％ＣＨ3ＣＮの勾配；８０分から１００分が０％Ｈ2Ｏ／１００％ＣＨ3ＣＮの勾配；１００分から１０１分が０％Ｈ2Ｏ／１００％ＣＨ3ＣＮの勾配；及び１０１分が９５％Ｈ2Ｏ／５％ＣＨ3ＣＮであり、流速は１ｍＬ／分であった。ＨＰＬＣは、Ｗａｔｅｒｓ Ａｌｌｉａｎｃｅ２７９５ＨＴで行われ、スペクトルは、Ｗａｔｅｒｓ Ｐｈｏｔｏ Ｄｉｏｄｅ Ａｒｒａｙ２９９６によって１９０から４００ｎｍで測定された。２００ｎｍで測定されたスペクトルに示すように、変動係数８％で、保持時間２．８９６、５．１１１、１２．９４４、及び４６．６０３分に鋭いピークがあった。
The result of 50 μl HPLC of fraction 3 is shown in FIG. The HPLC was a Waters Sphersorb (TM) -ODS2 column with an inner diameter (ID) of 4.6 mm, a length (L) of 250 mm and a particle size of 5 μm, and the mobile phase was 95% H 2 O / 5% CH 3 CN; 5 to 30 minutes 95% H 2 O / 5% CH 3 CN gradient; 30 to 80 minutes 75% H 2 O / 25% CH 3 CN gradient; 80 to 100
実施例２
軟膏の調製
本発明による軟膏を表２に例示する。
Example 2
Ointment Preparation Ointments according to the present invention are illustrated in Table 2.
本発明の軟膏は、当業者によって通常使用されるいかなる従来の方法を用いても調製することができる。 The ointment of the present invention can be prepared using any conventional method commonly used by those skilled in the art.
実施例３
ウサギにおける全層皮膚創傷に対する効果
動物の処置：
体重２．９〜３．１ｋｇの雌性成獣ニュージランドホワイトウサギ、５羽、を動物モデルとして用いた。各ウサギは、単独でケージに入れ、固形飼料及び水で飼育した。ウサギは、塩酸ケタミン２５〜４０ｍｇ／ｋｇで麻酔した。各ウサギの背中を入念に毛刈りし、６カ所の全層皮膚（各領域は、１．５×１．５ｃｍ2）を切除した。ここで、全層皮膚は、表皮、真皮及び皮筋層を含む。
Example 3
Effect of animal treatment on full thickness skin wounds in rabbits:
Five adult female New Zealand white rabbits weighing 2.9-3.1 kg were used as animal models. Each rabbit was singly caged and fed with solid feed and water. Rabbits were anesthetized with 25-40 mg / kg ketamine hydrochloride. Each rabbit's back was carefully shaved, and 6 layers of full-thickness skin (each area was 1.5 × 1.5 cm 2 ) were excised. Here, full-thickness skin includes the epidermis, dermis, and skin muscle layer.
投与及び頻度：
各ウサギの創傷６領域を、画分１〜６をそれぞれ含む軟膏で、１日２回治療した。軟膏は、０．３５ｇ／ｃｍ2の量を局所に塗布した。
Administration and frequency:
Six rabbit wounds were treated twice daily with an ointment containing fractions 1-6, respectively. The ointment was applied topically in an amount of 0.35 g / cm 2 .
創傷の臨床的評価：
創傷は、毎日観察し、創傷治癒の過程を記録するため、光、焦点距離及び口径の同一条件下で、週に３回、参照用スケールと共に写真撮影した。これに加えて、創傷の大きさも測定した。その結果を表３に示す。
Clinical evaluation of wounds:
Wounds were observed daily and photographed with a reference scale three times a week under the same conditions of light, focal length and caliber to record the process of wound healing. In addition, the size of the wound was also measured. The results are shown in Table 3.
上記の結果は、創傷が（図３に示すように）全て治癒したことを示している。本発明による賦形剤（画分５を含む軟膏）も、又、全層皮膚創傷の治療に効果を有していた。 The above results indicate that all the wounds were healed (as shown in FIG. 3). The excipient according to the invention (ointment containing fraction 5) also had an effect on the treatment of full thickness skin wounds.
体毛増殖の評価：
新しい体毛の長さを測定した。画分１〜６を含む軟膏で治療した全ての創傷の体毛は、平均約１．７６ｃｍの長さであり（図４に示すように）、治療しない領域のそれは平均０．１〜０．２ｃｍであった。
Evaluation of hair growth:
The length of new hair was measured. The body hair of all wounds treated with an ointment containing fractions 1-6 averages about 1.76 cm long (as shown in FIG. 4), and that of the untreated area averages 0.1-0.2 cm. Met.
結果は、本発明による揮発性画分が毛嚢欠損を修復する能力を有し、体毛増殖を促進した。これに加えて、本発明による賦形剤も、又、体毛増殖の増強に効果的であった。 The results showed that the volatile fraction according to the present invention had the ability to repair hair follicle defects and promoted hair growth. In addition to this, the excipient according to the invention was also effective in enhancing hair growth.
実施例４
ラットにおける全層皮膚創傷に対する効果
動物の処置：
雄性、成獣スプラーグ−ドーリーラットは、台湾国立Ｃｈｅｎｇ Ｋｕｎｇ大学実験動物センターから購入した。体重３００〜３５０ｇのラットは、塩酸ケタミン２５〜４０ｍｇ／ｋｇで麻酔した。各ラットの背中を電気クリッパで入念に毛刈りした。各ラットは、単独でケージに入れ、固形飼料及び水で飼育した。各ラットについて、２．５×２．５ｃｍ2の領域の全層皮膚を切除した。ここで、全層皮膚は、表皮、真皮、及び皮筋層を含む［Ｂ．Ｈａｆｅｍａｎｎら、真皮の組織工学用コラーゲン／エラスチン膜の使用、Ｂｕｒｎｓ．１９９９；２５：３７３−３８４；Ｈ．−Ｊ．Ｗａｎｇら、広範に拡張したメッシュの自己由来の中間層皮膚移植片増殖を増強するブタ真皮鋳型の使用、Ｊ．Ｔｒａｕｍａ．１９９７、２月；４２（２）：１７７−１８２］。
Example 4
Effect of animals on full thickness skin wounds in rats Treatment:
Male, adult Sprague-Dawley rats were purchased from Cheng Kung University Experimental Animal Center, National Taiwan. Rats weighing 300-350 g were anesthetized with 25-40 mg / kg ketamine hydrochloride. Each rat's back was carefully shaved with an electric clipper. Each rat was placed in a cage alone and fed with chow and water. For each rat, full thickness skin of a 2.5 × 2.5 cm 2 area was excised. Here, full-thickness skin includes the epidermis, dermis, and cutaneous muscle layer [B. Hafemann et al., Use of collagen / elastin membranes for tissue engineering of the dermis, Burns. 1999; 25: 373-384; -J. Wang et al., Use of Porcine Dermal Template to Enhance Widely Expanded Mesh Self-Derived Interlayer Skin Graft Growth. Trauma. 1997, February; 42 (2): 177-182].
投与及び頻度：
２実験群のラットを、実施例２で用いられた画分７及び８を含む軟膏で治療し、各群はラット３匹を含んでいた。医薬組成物を、０．３５ｇ／ｃｍ2及び高さ０．１〜０．２ｃｍの量で、創傷に局所塗布した。全ての創傷は、滅菌ガーゼ及び弾性包帯により手当てし、包帯を毎日交換した。
Administration and frequency:
Two experimental groups of rats were treated with the
実施例３と類似の評価方法を用い、創傷の大きさを測定した。結果を表４に示す。 Using an evaluation method similar to that in Example 3, the size of the wound was measured. The results are shown in Table 4.
画分７及び８を含む軟膏で治療した創傷は治癒した。特に、画分７を含む医薬組成物で治療した創傷領域はより速やかに減少した。
Wounds treated with an
実施例５
深部中間層皮膚熱傷の治癒に対する効果
動物の処置：
雌性、成獣スプラーグ−ドーリーラットは、台湾国立Ｃｈｅｎｇ Ｋｕｎｇ大学実験動物センターから購入した。体重２３０〜２８０ｇのラット９匹は、塩酸ケタミン２５〜４０ｍｇ／ｋｇで麻酔した。各ラットの背中を電気クリッパで、背中の中間部の予定した創傷領域を入念に毛刈りした。各ラットは、単独でケージに入れ、固形飼料及び水で飼育した。ラットの熱傷は、Ｗａｌｋｅｒ ａｎｄ Ｍａｓｏｎ［Ｚ−Ｒ．Ｇａｏら、皮膚ドナー部位及び深部中間層皮膚熱傷用の創傷用手当用品としてのブタ真皮コラーゲン、Ｂｕｒｎｓ．１９９２；１８（６）：４９２−４９６］によって記載された方法を用いて、背部皮膚表面を、４×４ｃｍ2の領域で深部中間層皮膚熱傷を生じるように設計された鋳型を通して、７５℃の水に１５秒間露出して生成させた。
Example 5
Effect on the healing of deep interlayer skin burns Animal treatment:
Female, adult Sprague-Dawley rats were purchased from the Cheng Kung University Experimental Animal Center, National Taiwan. Nine rats weighing 230-280 g were anesthetized with 25-40 mg / kg ketamine hydrochloride. Each rat's back was carefully clipped with an electric clipper and a planned wound area in the middle of the back. Each rat was placed in a cage alone and fed with chow and water. Rat burns were caused by Walker and Mason [ZR. Gao et al., Porcine dermal collagen as a wound dressing for skin donor sites and deep interlayer skin burns, Burns. 1992; 18 (6): 492-496], the dorsal skin surface was passed through a mold designed to produce deep interlayer skin burns in the 4 × 4 cm 2 area at 75 ° C. It was formed by exposure to water for 15 seconds.
投与及び頻度：
ラットの実験群（Ｇ３）を、アルコール含量を３％とした以外は実施例２と同様にして、画分３と同じように画分９を含む本発明に記載の医薬組成物で治療した。ラットの、無治療の１つの対照群（Ｇ１）とスルファジアジン銀（ＳＷＩＳＳ ＰＨＡＲＭＡＣＥＵＴＩＣＡＬ ＣＯ．，ＬＴＤ．）で治療した１つの比較群（Ｇ２）を用いた。各群はラット３匹を含んでいた。薬物は、体毛が増殖するまで、１日２回、０．３５ｇ／ｃｍ2の量を創傷に局所塗布した。
Administration and frequency:
The experimental group of rats (G3) was treated with the pharmaceutical composition according to the invention comprising fraction 9 in the same way as fraction 3 in the same manner as in example 2, except that the alcohol content was 3%. One untreated control group (G1) and one comparative group (G2) of rats treated with silver sulfadiazine (SWISS PHARMACEUTICAL CO., LTD.) Were used. Each group contained 3 rats. The drug was topically applied to the wound in an amount of 0.35 g / cm 2 twice a day until the hair grew.
創傷の臨床的評価：
実施例３と類似した評価方法を用い、又、創傷の大きさを測定した。結果を表５に示す。
Clinical evaluation of wounds:
An evaluation method similar to that in Example 3 was used, and the size of the wound was measured. The results are shown in Table 5.
創傷治癒の進行において、本発明による軟膏で治療した創傷は瘢痕なしに治癒し、皮膚の色は正常と同じであった。しかしながら、スルファジアジン銀で治療した創傷には瘢痕が生じた。更に、ラットの創傷は、第３７日目においても創傷領域の２．８６％に瘢痕が残っていた従来のスルファジアジン銀で治療したよりも早く、第１８日目で治癒した。
In the course of wound healing, the wound treated with the ointment according to the invention healed without scarring and the skin color was the same as normal. However, scars occurred in wounds treated with silver sulfadiazine. Furthermore, rat wounds healed on day 18 earlier than on conventional sulfadiazine silver treatment, which remained scarred in 2.86% of the wound area on
新しい体毛増殖の成績（スコア）も評価し、記録した。新しい体毛増殖スコアは、以下のように定義された：「−２」は、幾分痂皮及び紅斑があり、新しい体毛増殖はないもの；「−１」は、幾分紅斑があり、新しい体毛増殖はないもの；「０」は、正常の皮膚で、新しい体毛増殖はないもの；「＋１」は、幾分新しい体毛増殖があるもの；及び「＋２」は、創傷領域のおよそ５０％の新しい体毛増殖があるもの。 New hair growth performance (score) was also evaluated and recorded. The new hair growth score was defined as follows: "-2" is somewhat scab and erythema, no new hair growth; "-1" is somewhat erythema and new hair "0" is normal skin, no new hair growth; "+1" is somewhat new hair growth; and "+2" is approximately 50% new in the wound area Those with hair growth.
新しい体毛増殖スコアを、表６に列挙する。 New hair growth scores are listed in Table 6.
表６に示したように、本発明の軟膏は、従来のスルファジアジン銀よりも新しい体毛増殖により効果的であった。新しい体毛増殖は、本発明による医薬組成物で治療した第１８日目に観察され、新しい体毛増殖の５０％は第３５日目に観察された。本発明による医薬組成物で治療した創傷には、紅斑も痂皮も認められなかった。しかしながら、従来の薬物で治療した場合は、新しい体毛増殖は見られなかった。 As shown in Table 6, the ointment of the present invention was more effective at growing new hair than conventional sulfadiazine silver. New hair growth was observed on day 18 treated with the pharmaceutical composition according to the invention, and 50% of the new hair growth was observed on day 35. No erythema or scab was found in the wound treated with the pharmaceutical composition according to the present invention. However, no new hair growth was seen when treated with conventional drugs.
皮膚の組織学的評価：
創傷が完全に治癒した後に、ラットを麻酔して各ラットの皮膚組織を採取し、Ｂ．Ｈａｆｅｍａｎｎら（Ｂ．Ｈａｆｅｍａｎｎら、真皮の組織工学用コラーゲン／エラスチン膜の使用、Ｂｕｒｎｓ．１９９９；２５：３７３−３８４）によって記載された評価方法を使用して組織学的試験を行った。試料を採取し、ホルムアルデヒド中に貯蔵した。各検体は、パラフィンブロックに包埋し、薄切片を調製し、ヘマトキシリン・エオジン法を用いて染色した。検体は、光学顕微鏡及び透過型電子顕微鏡で観察した。
Histological evaluation of the skin:
After the wound has healed completely, the rats are anesthetized and the skin tissue of each rat is collected; Histological examination was performed using the evaluation method described by Hafemann et al. (B. Hafemann et al., Use of collagen / elastin membrane for tissue engineering of the dermis, Burns. 1999; 25: 373-384). Samples were taken and stored in formaldehyde. Each specimen was embedded in a paraffin block, a thin section was prepared, and stained using the hematoxylin-eosin method. The specimen was observed with an optical microscope and a transmission electron microscope.
組織学的試験の結果は、無治療のラット上皮組織の厚さが（図５に示すように）平らでなく、顆粒層が厚くなっていることを示した。線維層は、深部皮膚に見られた。従来のスルファジアジン銀で治療したラット群でも（図６に示すように）、又、不均一な厚さの上皮組織を有することが示された。線維層は深部皮膚に見られ、毛嚢は崩壊していた。一方、本発明による軟膏で治療した創傷は（図７に示すように）、正常であるように見え、多数の毛嚢が再生していた。線維層は、真皮の表面領域に見られた。 The results of histological examination showed that the untreated rat epithelial tissue was not flat (as shown in FIG. 5) and the granule layer was thickened. A fibrous layer was found in the deep skin. A group of rats treated with conventional silver sulfadiazine (as shown in FIG. 6) was also shown to have non-uniform thickness of epithelial tissue. The fibrous layer was found in the deep skin and the hair follicles were disintegrated. On the other hand, the wound treated with the ointment according to the present invention (as shown in FIG. 7) appeared to be normal and many hair follicles had regenerated. Fibrous layers were found in the surface area of the dermis.
実施例６
ラットにおける大領域の全層皮膚傷害に対する効果
動物の処置：
雄性、成獣スプラーグ−ドーリーラットは、台湾国立Ｃｈｅｎｇ Ｋｕｎｇ大学実験動物センターから購入した。体重３００〜３５０ｇのラットは、塩酸ケタミン２５〜４０ｍｇ／ｋｇで麻酔した。各ラットの背中を電気クリッパで入念に毛刈りした。各ラットは、単独でケージに入れ、固形飼料及び水で飼育した。各ラットについて、４．０×４．０ｃｍ2の領域の全層皮膚を切除した。ここで、全層皮膚は表皮、真皮、及び皮筋層を含む。
Example 6
Effect on large area full thickness skin injury in rats Treatment of animals:
Male, adult Sprague-Dawley rats were purchased from Cheng Kung University Experimental Animal Center, National Taiwan. Rats weighing 300-350 g were anesthetized with 25-40 mg / kg ketamine hydrochloride. Each rat's back was carefully shaved with an electric clipper. Each rat was placed in a cage alone and fed with chow and water. For each rat, the full thickness skin of a 4.0 × 4.0 cm 2 area was excised. Here, full-thickness skin includes the epidermis, dermis, and cutaneous muscle layer.
投与及び頻度：
ラットの実験群は、実施例５の画分９を含む軟膏で治療された。スルファジアジン銀（ＳＷＩＳＳ ＰＨＡＲＭＡＣＥＵＴＩＣＡＬ ＣＯ．，ＬＴＤ．）で治療したラットの１つの比較群（Ｇ２）も、又、使用された。各群は、ラット５匹を含んでいた。軟膏は、０．７５ｇ／ｃｍ2及び高さ０．１〜０．２ｃｍの量で、創傷に局所塗布した。全ての創傷は、滅菌ガーゼ及び弾性包帯により手当てし、包帯を毎日交換した。
Administration and frequency:
An experimental group of rats was treated with an ointment containing fraction 9 of Example 5. One comparative group (G2) of rats treated with silver sulfadiazine (SWISS PHARMACEUTICAL CO., LTD.) Was also used. Each group contained 5 rats. The ointment was topically applied to the wound in an amount of 0.75 g / cm 2 and a height of 0.1-0.2 cm. All wounds were treated with sterile gauze and elastic bandages and the bandages were changed daily.
実施例３と類似の評価方法を用い、創傷の大きさを測定した。結果を表７に示す。 Using an evaluation method similar to that in Example 3, the size of the wound was measured. The results are shown in Table 7.
創傷治癒の進行において、本発明による軟膏で治療した創傷は、アレルギーなしに治癒した。しかしながら、スルファジアジン銀で治療した創傷には瘢痕が生じた。更に、スルファジアジン銀で治療した創傷には、重篤なアレルギーが見られた。本発明による医薬組成物で治療したラットの治癒率（約２５〜３１日）は、スルファジアジン銀で治療した（約３７日）よりも早かった。 In the course of wound healing, the wound treated with the ointment according to the invention healed without allergies. However, scars occurred in wounds treated with silver sulfadiazine. In addition, severe allergies were seen in wounds treated with silver sulfadiazine. The cure rate of rats treated with the pharmaceutical composition according to the invention (about 25-31 days) was faster than that treated with silver sulfadiazine (about 37 days).
皮膚の組織学的評価：
組織学的評価方法は、実施例５に記載された通りである。
Histological evaluation of the skin:
The histological evaluation method is as described in Example 5.
第４５日目に行った組織学的試験の結果は、従来のスルファジアジン銀で治療した創傷は、深部真皮に線維層を有していることが観察されることを示した。いくつかの表皮細胞は死亡し、胼胝を形成していた。慢性炎症及び好酸球浸潤の両者が真皮に観察された。いくらかの線維も、又、創傷から０．５ｃｍ離れたところに形成していた。 The results of a histological study performed on day 45 showed that wounds treated with conventional silver sulfadiazine were observed to have a fibrous layer in the deep dermis. Some epidermal cells died and formed folds. Both chronic inflammation and eosinophil infiltration were observed in the dermis. Some fibers also formed 0.5 cm away from the wound.
本発明による軟膏で治療したラットの群では、新生した上皮は均一で、多数の毛嚢が生成した。ほんの少しの線維が形成したのみであった。 In the group of rats treated with the ointment according to the invention, the new epithelium was uniform and numerous hair follicles formed. Only a few fibers formed.
第９５日目に行った組織学的試験の結果は、従来のスルファジアジン銀で治療した創傷には、不均一な上皮層を有していることが観察される（図８に示すように）ことを示した。線維層が深部真皮に形成された。少量の毛嚢が生成した。 The results of the histological examination performed on day 95 show that the wound treated with conventional silver sulfadiazine has a heterogeneous epithelial layer (as shown in FIG. 8). showed that. A fibrous layer was formed in the deep dermis. A small amount of hair follicles formed.
図９に示すように、本発明による軟膏で治療したラットの群では、新生した上皮は均一で、多数の毛嚢が生成した。 As shown in FIG. 9, in the group of rats treated with the ointment according to the present invention, the newly formed epithelium was uniform and a large number of hair follicles were formed.
実施例７
局所電子照射皮膚傷害に対する効果
動物の処置：
雌性、成獣スプラーグ−ドーリーラットは、台湾国立Ｃｈｅｎｇ Ｋｕｎｇ大学実験動物センターから購入した。体重２５０〜３００ｇのラットは、単独でケージに入れ、固形飼料及び水で飼育した。
Example 7
Effect on local electron irradiation skin injury Animal treatment:
Female, adult Sprague-Dawley rats were purchased from the Cheng Kung University Experimental Animal Center, National Taiwan. Rats weighing 250-300 g were singly caged and fed with solid feed and water.
ラットの照射は、Ｊｏｈｎ Ｅ．Ｍｏｕｌｄｅｒ ａｎｄ Ｊａｍｅｓ Ｊ．Ｆｉｓｈｅｒ［ラット皮膚の放射線に対する反応：各Ｎｏ．の画分の役割と総処置時間、Ｃａｎｃｅｒ、６月、１９７６年、３７（６）：２７６２−２７６７］によって記載されたのと類似の方法を用いて行われた。ラットは、照射前に、４０〜５０ｍｇ／ｋｇのペントバルビタールナトリウム塩（ＴＣＩ．東京）の腹腔内投与により麻酔した。臀筋領域にわたる皮膚は完全に体毛を剃り落とし、照射前に、右と左の臀筋上の所望の２×２ｃｍ2の照射領域に印をつけた。線形加速器（Ｖａｒｉａｎ Ａｓｓｏｃｉａｔｅｓ，Ｉｎｃ．，ＣＡ，Ｕ．Ｓ．Ａ．）によって発生した６ＭｅＶの電子ビームを照射に用いた。 Rats were irradiated by John E. Moulder and James J.M. Fisher [Reaction of rat skin to radiation: No. Of the fractions and total treatment time, Cancer, June, 1976, 37 (6): 2762-2767]. Rats were anesthetized by intraperitoneal administration of 40-50 mg / kg pentobarbital sodium salt (TCI. Tokyo) prior to irradiation. The skin over the gluteal area was completely shaved and the desired 2 × 2 cm 2 irradiated areas on the right and left gluteal muscles were marked before irradiation. A 6 MeV electron beam generated by a linear accelerator (Varian Associates, Inc., CA, USA) was used for irradiation.
分割照射及び単回照射を試験した。分割照射では、ラット３匹に対し、右及び左の臀筋に照射した。照射は、毎週２回、５週間行い、総線量は各領域につき３７．５Ｇｙであった。 Split irradiation and single irradiation were tested. In split irradiation, the right and left gluteal muscles were irradiated to 3 rats. Irradiation was performed twice weekly for 5 weeks, and the total dose was 37.5 Gy for each area.
投与及び頻度：
分割照射における照射傷害は、照射の後に１日１回、投与量０．２５ｇ／ｃｍ2の実施例５で用いられた画分９を含む軟膏で治療し、単回照射における照射傷害は、一日二回治療した。治療なしのラットは、対照（Ｇ２）として使用した。照射しないラットの一つのブランク群（Ｇ３）は、ブランクとした。
Administration and frequency:
Irradiation injury in fractionated irradiation was treated with an ointment containing fraction 9 used in Example 5 at a dose of 0.25 g / cm 2 once a day after irradiation. Treated twice a day. Rats without treatment were used as controls (G2). One blank group (G3) of rats that were not irradiated was blank.
臨床評価
皮膚反応スコア［Ｙｏｓｈｉｎａｏ Ａｂｅ ａｎｄ Ｍｕｎｅｙａｓｕ Ｕｒａｎｏ、多回、一日二回照射後のマウスの分割照射側に依存した急性皮膚反応、Ｊ．Ｒａｄｉａｔｉｏｎ Ｏｎｃｏｌｏｇｙ Ｂｉｏｌ． Ｐｈｙｓ．１９９０年２月；１８（２）：３５９−３６４；Ｙｕ−Ｊｅｎ Ｃｈｅｎら、ラットにおける急性放射線皮膚炎に対するテトランドリン及びツボクサ抽出物の効果、Ｂｉｏｌ．Ｐｈａｒｍ．Ｂｕｌｌ．１９９９年７月；２２（７）：７０３−７０６］を評価し、照射後３０日間記録した。皮膚反応スコアは、以下のように定義された：「０」は正常皮膚を示し；「０．５」は照射した領域の５０％未満に軽い脱毛が起こる、又は体毛の脱色が起こることを示し；「１．０」は照射した領域の約５０％に脱毛が起こることを示し；「１．５」は照射した領域の５０％以上に脱毛が起こり、いくつかの脱毛は紅皮を伴うことを示し；「２．０」は完全な脱毛及び照射した領域の５０％未満に乾燥した落屑が起こることを示し；「２．５」は照射した領域の５０％以上に乾燥した落屑が起こることを示し；「３．０」は幾分湿潤した落屑が起こることを示し；「３．５」は領域の殆どに湿潤した落屑が起こることを示す。
Clinical evaluation Skin reaction score [Yoshina Abe and Muneyasu Urano, acute skin reaction depending on the divided irradiation side of mice after irradiation twice a day, J. Radiation Oncology Biol. Phys. 1990 1990; 18 (2): 359-364; Yu-Jen Chen et al., Effect of tetrandrine and cinnamon extract on acute radiation dermatitis in rats, Biol. Pharm. Bull. 1999 1999; 22 (7): 703-706] and was recorded for 30 days after irradiation. The skin reaction score was defined as follows: “0” indicates normal skin; “0.5” indicates light hair loss or hair loss occurs in less than 50% of the irradiated area. "1.0" indicates that hair loss occurs in about 50% of the irradiated area; "1.5" indicates that hair loss occurs in more than 50% of the irradiated area, and some hair loss is associated with red skin "2.0" indicates complete depilation and dry desquamation occurs in less than 50% of the irradiated area; "2.5" indicates dry desquamation occurs in 50% or more of the irradiated area. “3.0” indicates that some moist desquamation occurs; “3.5” indicates that moist desquamation occurs in most of the area.
皮膚反応スコアを評価した。結果を表８に示す。 Skin reaction score was evaluated. The results are shown in Table 8.
Ｇ１では、裂傷初期と僅かの体毛損傷が創傷に見られ、皮膚はアレルギーを示さなかった。
結果は、分割照射試験の場合と同様であった。
In G1, early wounds and slight hair damage were seen in the wounds, and the skin showed no allergy.
The result was the same as in the case of the split irradiation test.
皮膚の組織学的評価：
実施例５におけると同様の評価方法を使用した。
Histological evaluation of the skin:
The same evaluation method as in Example 5 was used.
図１０に示すように、Ｇ１において、表皮細胞は、多層の上皮細胞を覆っていることが見出された。又、多数の毛嚢が再生された。 As shown in FIG. 10, it was found that in G1, epidermal cells covered multiple layers of epithelial cells. Many hair follicles were regenerated.
図１１に示すように、Ｇ２において、ほんの少量の毛嚢が観察された。 As shown in FIG. 11, only a small amount of hair follicle was observed in G2.
図１２に示すように、Ｇ３において、組織は対照として使用した。 As shown in FIG. 12, in G3, tissue was used as a control.
治療した創傷の組織は密であったが、無治療のものは粗であった。 Treated wound tissue was dense, but untreated was rough.
実施例８
ウサギ耳における皮膚炎に対する効果
ウサギ耳における皮膚炎は、図１３に示すように、発赤、腫れ、炎症、潰瘍及び出血を有することが見出された。
Example 8
Effect on dermatitis in rabbit ears Dermatitis in rabbit ears was found to have redness, swelling, inflammation, ulcers and bleeding as shown in FIG.
アルコール含量を０％とした以外は画分３に類似した画分１０を用いた以外は実施例２と同じようにして、本発明による軟膏を創傷に３日間塗布した。 The ointment according to the present invention was applied to the wound for 3 days in the same manner as in Example 2 except that fraction 10 similar to fraction 3 was used except that the alcohol content was 0%.
第１日目は、潰瘍が消失し、発赤が軽減した。毛嚢が図１４に示すよう現れた。 On the first day, the ulcer disappeared and redness was reduced. A hair follicle appeared as shown in FIG.
第２日目は、皮膚組織が回復し、ほんの軽い発赤が観察された。毛嚢は図１５に示すよう明らかであった。 On the second day, skin tissue recovered and only slight redness was observed. The hair follicle was evident as shown in FIG.
第３日目は、皮膚組織及び色彩は正常であった。体毛が、図１６に示すように毛嚢に生えてきた。 On the third day, skin tissue and color were normal. Body hair has grown in the hair follicle as shown in FIG.
実施例９
糖尿病性創傷に対する効果
６８歳の男性が糖尿病に２０年間かかっていた。彼の左足の足指は切断された。創傷は、図１７に示すように重篤な壊疽変性を示した。彼の右足の足指は切断されたが、しかし創傷治癒は図１８に示すように良好でなかった。
Example 9
Effect on Diabetic Wound A 68 year old man had diabetes for 20 years. His left toe was severed. The wound showed severe gangrene degeneration as shown in FIG. His right toe was severed, but wound healing was not good as shown in FIG.
画分９を含む軟膏が、創傷に１日２〜３回塗布された。 An ointment containing fraction 9 was applied to the wound 2-3 times a day.
図１９に示すように、２ヶ月間左足の治療の後、いくつかの肉芽組織が認められ、そして創傷は、それ以上壊死の進行がなく治癒し始めた。 As shown in FIG. 19, after 2 months of treatment of the left foot, some granulation tissue was observed and the wound began to heal without further progression of necrosis.
５ヶ月間左足の治療の後、図２０に示すように、創傷は瘢痕なしに完全に治癒した。図２１に示すように、右足も又、瘢痕なしに完全に治癒した。 After treatment of the left foot for 5 months, the wound healed completely without scarring, as shown in FIG. As shown in FIG. 21, the right foot also healed completely without scarring.
実施例１０
慢性うっ血性潰瘍に対する効果
８０歳の男性の両下腿は、図２２に示すように色素過剰症及びうっ血性潰瘍に２０年間かかっていた。
Example 10
Effect on Chronic Congestive Ulcers The lower leg of an 80-year-old man suffered from hyperpigmentation and congestive ulcer for 20 years as shown in FIG.
画分９を含む軟膏を、創傷に１日２〜３回塗布した。 An ointment containing fraction 9 was applied to the wound 2-3 times a day.
図２３に示すように、２週間足を治療した後、創傷は治癒し始めた。 As shown in FIG. 23, the wound began to heal after treating the paw for 2 weeks.
図２４に示すように、４週間足を治療した後、創傷は瘢痕なしに完全に治癒した。 As shown in FIG. 24, after treating the foot for 4 weeks, the wound healed completely without scarring.
実施例１１
斑状出血に対する効果
女性患者は、図２５に示すように、事故に起因する斑状出血を有していた。
Example 11
Effect on ecchymosis The female patient had ecchymosis due to an accident as shown in FIG.
創傷は、３日の間、１日１回、冷湿布で予備治療され、その後画分９を含む軟膏を１日２〜３回塗布した。 The wound was pretreated with cold compress once a day for 3 days, after which an ointment containing fraction 9 was applied 2-3 times a day.
斑状出血は消失し、創傷は図２６に示すように、治療後３日目に治癒した。自動車事故の前から既に存在していた瘢痕の色も又、図２７に示すように、７日間の治療の後、平らになり薄くなった。 The ecchymosis disappeared and the wound healed 3 days after treatment as shown in FIG. The scar color already present before the car accident also flattened and faded after 7 days of treatment, as shown in FIG.
実施例１２
擦過傷に対する効果
女性患者は、図２８に示すように事故に起因する擦過傷を有していた。
Example 12
Effect on Abrasion The female patient had an abrasion due to an accident as shown in FIG.
画分９を含む軟膏を、創傷に１日２〜３回塗布した。 An ointment containing fraction 9 was applied to the wound 2-3 times a day.
創傷は、図２９に示すように、７日間の治療の後治癒した。 The wound healed after 7 days of treatment, as shown in FIG.
実施例１３
縫合創傷に対する効果
男性患者は、３ｃｍの裂傷と２０針の縫合の人中の重篤な裂傷、０．８ｃｍの裂傷と４針の縫合の鼻梁、及び１ｃｍの裂傷と６針の縫合及び１．５ｃｍの裂傷と８針の縫合の上唇を有していた。
Example 13
Effect on Suture Wounds Male patients have severe lacerations in humans with 3 cm lacerations and 20 needle sutures, 0.8 cm lacerations and 4 needle suture nasal bridges, and 1 cm lacerations and 6 needle sutures. It had a 5 cm laceration and an upper lip of 8 stitches.
画分９を含む軟膏を、創傷に１日２〜３回塗布した。 An ointment containing fraction 9 was applied to the wound 2-3 times a day.
創傷は、図３０に示すように、瘢痕なしに完全に治癒した。 The wound healed completely without scarring as shown in FIG.
実施例１４
アトピー性皮膚炎に対する効果
４．５歳の女性は、慢性アトピー性皮膚炎を有していた。頚部及び上胸部の皮膚は、図３１に示すように、長期間の局所コルチコステロイド療法の後に、痒み、発赤、腫張、かさかさの状態、湿潤状態、痂皮状態及びかさぶた状態であった。
Example 14
Effect on Atopic Dermatitis A 4.5 year old woman had chronic atopic dermatitis. As shown in FIG. 31, the skin of the neck and upper chest was itchy, reddened, swollen, bulky, wet, crusted, and scabted after long-term topical corticosteroid therapy.
画分１０を含む軟膏を、創傷に１日２〜３回塗布した。 An ointment containing fraction 10 was applied to the wound 2-3 times a day.
図３２に示すように、１週間の治療の後、痒み、発赤、腫張、かさかさの状態、湿潤状態、痂皮状態及びかさぶた状態は、軽減し減少した。 As shown in FIG. 32, after one week of treatment, itching, redness, swelling, bulkiness, wetness, crusting and scab conditions alleviated and decreased.
図３３に示すように、４週間の治療の後、皮膚病変は殆ど沈静化した。 As shown in FIG. 33, the skin lesions almost subsided after 4 weeks of treatment.
図３４に示すように、６週間の治療の後、皮膚病変は、後のリバウンド効果もなく完全に治癒した。 As shown in FIG. 34, after 6 weeks of treatment, the skin lesions were completely healed without a subsequent rebound effect.
本発明の実施態様が例示され記載されているが、種々の変形及び改良は、当業者によって行うことができる。本発明の実施態様は、それ故に、例示的に記載されたものであり、限定的な意味で記載されたものではない。本発明は、例示したような特定の形態に限定されるものではなく、本発明の精神及び範囲から逸脱していない全ての変形は、付属する特許請求の範囲に定義された範囲内にある。 While embodiments of the invention have been illustrated and described, various modifications and improvements can be made by those skilled in the art. Embodiments of the present invention are therefore described by way of example and not in a limiting sense. The invention is not limited to the specific forms as illustrated, and all modifications that do not depart from the spirit and scope of the invention are within the scope defined by the appended claims.
Claims (49)
（ａ）約１５重量％未満の濃度のアルコールを含有するアルコール溶液中、又は水中に種子の粗抽出物を供給し；そして
（ｂ）約１気圧未満の気圧及び約１１０℃未満の温度で、粗抽出物を蒸発させて揮発性画分を得る；
を含むプロセスによって調製され；且つ、
内径（Ｉ．Ｄ．）４．６ｍｍ、長さ（Ｌ）２５０ｍｍ及び粒子径５μｍのＷａｔｅｒｓ Ｓｐｈｅｒｓｏｒｂ（ＴＭ）−ＯＤＳ２カラムを用いた高速液体クロマトグラフィー（ＨＰＣＬ）を通じて２００ｎｍで測定した、該揮発性画分５０μｌのスペクトルが、変動係数８％で、保持時間２．９１０、５．１９０、１３．１９０及び５０．８１５分にピークを有し、ここで、移動相は、０から５分が９５％Ｈ2Ｏ／５％ＣＨ3ＣＮ；５分から３０分が９５％Ｈ2Ｏ／５％ＣＨ3ＣＮの勾配；３０分から８０分が７５％Ｈ2Ｏ／２５％ＣＨ3ＣＮの勾配；８０分から１００分が０％Ｈ2Ｏ／１００％ＣＨ3ＣＮの勾配；１００分から１０１分が０％Ｈ2Ｏ／１００％ＣＨ3ＣＮの勾配；及び１０１分が９５％Ｈ2Ｏ／５％ＣＨ3ＣＮであり、流速が１ｍＬ／分である；
である揮発性画分。 Soybean [Glycine max (L) Merr. The volatile fraction from the seeds of the following steps:
(A) providing a crude extract of seeds in an alcohol solution containing alcohol at a concentration of less than about 15% by weight, or in water; and (b) at a pressure of less than about 1 atmosphere and a temperature of less than about 110 ° C. Evaporate the crude extract to obtain a volatile fraction;
Prepared by a process comprising: and
The volatile fraction measured at 200 nm through high performance liquid chromatography (HPCL) using a Waters Sphersorb (TM) -ODS2 column with an inner diameter (ID) of 4.6 mm, length (L) of 250 mm and particle size of 5 μm. The 50 μl minute spectrum has a coefficient of variation of 8% and peaks at retention times of 2.910, 5.190, 13.190 and 50.815 minutes, where the mobile phase is 95% from 0 to 5 minutes. H 2 O / 5% CH 3 CN; 5 to 30 minutes from 95% H 2 O / 5% CH 3 CN gradient; 30 to 80 minutes from 75% H 2 O / 25% CH 3 CN gradient; from 80 minutes 100 min 0% H 2 O / 100% CH 3 CN gradient of; gradient 100 minutes 101 minutes is 0% H 2 O / 100% CH 3 CN ; and 101 min 95% H 2 O / 5% CH 3 CN, flow velocity A 1 mL / min;
Is a volatile fraction.
（ａ）約１５重量％未満の濃度のアルコールを含有するアルコール溶液中、又は水中に原料の粗抽出物を供給し；そして
（ｂ）約１気圧未満の気圧及び約１１０℃未満の温度で、粗抽出物を蒸発させて揮発性画分を得る；
を含むプロセスによって調製され；且つ、
内径（Ｉ．Ｄ．）４．６ｍｍ、長さ（Ｌ）２５０ｍｍ及び粒子径５μｍのＷａｔｅｒｓ Ｓｐｈｅｒｓｏｒｂ（ＴＭ）−ＯＤＳ２カラムを用いた高速液体クロマトグラフィー（ＨＰＣＬ）を通じて２００ｎｍで測定した、揮発性画分５０μｌのスペクトルが、変動係数８％で、保持時間２．９１０、５．１９０、１３．１９０、及び５０．８１５分にピークを有し、ここで、移動相は、０から５分が９５％Ｈ2Ｏ／５％ＣＨ3ＣＮ；５分から３０分が９５％Ｈ2Ｏ／５％ＣＨ3ＣＮの勾配；３０分から８０分が７５％Ｈ2Ｏ／２５％ＣＨ3ＣＮの勾配；８０分から１００分が０％Ｈ2Ｏ／１００％ＣＨ3ＣＮの勾配；１００分から１０１分が０％Ｈ2Ｏ／１００％ＣＨ3ＣＮの勾配；及び１０１分が９５％Ｈ2Ｏ／５％ＣＨ3ＣＮであり、流速が１ｍＬ／分である；
である揮発性画分。 Soybean [Glycine max (L) Merr. And a volatile fraction from a raw material comprising at least one selected from the group consisting of seeds of sesame (Sesamum indicum), seeds of mungbean (Phaseolus radiatas L.) and antibacterial herbs, Steps:
(A) supplying a crude extract of the raw material in an alcohol solution containing alcohol at a concentration of less than about 15% by weight or in water; and (b) at a pressure of less than about 1 atmosphere and a temperature of less than about 110 ° C. Evaporate the crude extract to obtain a volatile fraction;
Prepared by a process comprising: and
Volatile fraction measured at 200 nm through high performance liquid chromatography (HPCL) using a Waters Sphersorb (TM) -ODS2 column with an inner diameter (ID) of 4.6 mm, length (L) of 250 mm and particle size of 5 μm. The 50 μl spectrum has peaks at retention times 2.910, 5.190, 13.190, and 50.815 minutes with a coefficient of variation of 8%, where the mobile phase is 95% from 0 to 5 minutes. H 2 O / 5% CH 3 CN; 5 to 30 minutes from 95% H 2 O / 5% CH 3 CN gradient; 30 to 80 minutes from 75% H 2 O / 25% CH 3 CN gradient; from 80 minutes 100 min 0% H 2 O / 100% CH 3 CN gradient of; gradient 100 minutes 101 minutes is 0% H 2 O / 100% CH 3 CN ; and 101 min 95% H 2 O / 5% CH 3 CN, flow velocity A 1 mL / min;
Is a volatile fraction.
（ａ）約２％のアルコール中に原料の粗抽出物を供給し；そして
（ｂ）約１気圧未満の気圧及び約１１０℃未満の温度で、粗抽出物を蒸発させて揮発性画分を得る；
ステップを含むプロセスによって調製される方法。 A sufficient amount of about 70% soybean seed, about 20% sesame seed, about 5% effective to promote skin injury treatment, skin disease treatment, enhanced cell regeneration and hair growth. Skin injury or skin disease in a patient comprising applying to the patient's wound a composition comprising a volatile fraction from a raw material comprising dried rhizomes of palm leaves or yellowtail and about 5% yellowfin or dried buckwheat bark Wherein the volatile fraction is:
(A) feed the raw crude extract in about 2% alcohol; and (b) evaporate the crude extract at a pressure of less than about 1 atmosphere and a temperature of less than about 110 ° C. to produce a volatile fraction. obtain;
A method prepared by a process comprising steps.
（ａ）約２％のアルコール中に原料の粗抽出物を供給し；そして
（ｂ）約１気圧未満の気圧及び約１１０℃未満の温度で、粗抽出物を蒸発させて揮発性画分を得る；
ステップを含むプロセスによって調製され；且つ、
内径（Ｉ．Ｄ．）４．６ｍｍ、長さ（Ｌ）２５０ｍｍ及び粒子径５μｍのＷａｔｅｒｓ Ｓｐｈｅｒｓｏｒｂ（ＴＭ）−ＯＤＳ２カラムを用いた高速液体クロマトグラフィー（ＨＰＣＬ）を通じて２００ｎｍで測定した、揮発性画分５０μｌのスペクトルが、変動係数８％で、保持時間２．８９６、５．１１１、１２．９４４及び４６．６０３分にピークを有し、ここで、移動相は、０から５分が９５％Ｈ2Ｏ／５％ＣＨ3ＣＮ；５分から３０分が９５％Ｈ2Ｏ／５％ＣＨ3ＣＮの勾配；３０分から８０分が７５％Ｈ2Ｏ／２５％ＣＨ3ＣＮの勾配；８０分から１００分が０％Ｈ2Ｏ／１００％ＣＨ3ＣＮの勾配；１００分から１０１分が０％Ｈ2Ｏ／１００％ＣＨ3ＣＮの勾配；及び１０１分が９５％Ｈ2Ｏ／５％ＣＨ3ＣＮであり、流速が１ｍＬ／分である；
である揮発性画分。 Sufficient amount of about 53% soybean seed, about 20% sesame seed, about 17% effective to promote skin injury treatment, skin disease treatment, promote cell regeneration and hair growth Volatile fractions from a raw material comprising mungbean seeds, about 5% palm leaf yellow or dairy dry rhizome, and about 5% dried yellowfin or duck bark:
(A) feed the raw crude extract in about 2% alcohol; and (b) evaporate the crude extract at a pressure of less than about 1 atmosphere and a temperature of less than about 110 ° C. to produce a volatile fraction. obtain;
Prepared by a process comprising steps; and
Volatile fraction measured at 200 nm through high performance liquid chromatography (HPCL) using a Waters Sphersorb (TM) -ODS2 column with an inner diameter (ID) of 4.6 mm, length (L) of 250 mm and particle size of 5 μm. A 50 μl spectrum has peaks at retention times 2.896, 5.111, 12.944 and 46.603 minutes with a coefficient of variation of 8%, where the mobile phase is 95% H from 0 to 5 minutes. 2 O / 5% CH 3 CN; gradient from 5 to 30 minutes 95% H 2 O / 5% CH 3 CN; 30 to 80 minutes gradient from 75% H 2 O / 25% CH 3 CN; 80 minutes to 100 min 0% H 2 O / 100% CH 3 CN gradient; 100 min 101 min 0% H 2 O / 100% CH 3 CN gradient; and 101 min 95% H 2 O / 5% CH 3 CN And the flow rate is A mL / min;
Is a volatile fraction.
（ａ）約２％のアルコール中に原料の粗抽出物を供給し；そして
（ｂ）約１気圧未満の気圧及び約１１０℃未満の温度で、粗抽出物を蒸発させて揮発性画分を得る；
ステップを含むプロセスによって調製される方法。 From a source comprising about 80% soybean seed and about 20% mungbean seed in an amount sufficient to promote skin injury treatment, skin disease treatment, enhanced cell regeneration and hair growth. A method of treating a skin injury or skin disorder in a patient comprising applying a composition comprising a volatile fraction to a patient's wound; the volatile fraction comprising:
(A) feed the raw crude extract in about 2% alcohol; and (b) evaporate the crude extract at a pressure of less than about 1 atmosphere and a temperature of less than about 110 ° C. to produce a volatile fraction. obtain;
A method prepared by a process comprising steps.
（ａ）約２％のアルコール中に原料の粗抽出物を供給し；そして
（ｂ）約１気圧未満の気圧及び約１１０℃未満の温度で、粗抽出物を蒸発させて揮発性画分を得る；
ステップを含むプロセスによって調製される方法。 From a source comprising about 80% soybean seed and about 20% sesame seed in a sufficient amount effective to treat skin injury, skin disease, promote cell regeneration and hair growth. A method of treating skin injury or skin disease in a patient comprising applying a composition comprising a volatile fraction to a patient's wound; the volatile fraction comprising:
(A) feed the raw crude extract in about 2% alcohol; and (b) evaporate the crude extract at a pressure of less than about 1 atmosphere and a temperature of less than about 110 ° C. to produce a volatile fraction. obtain;
A method prepared by a process comprising steps.
（ａ）約２％のアルコール中に原料の粗抽出物を供給し；そして
（ｂ）約１気圧未満の気圧及び約１１０℃未満の温度で、粗抽出物を蒸発させて揮発性画分を得る；
ステップを含むプロセスによって調製される方法。 A sufficient amount of about 90% soybean seeds and about 10% palm leaves Daio or Daiou dry rhizome effective to promote skin injury treatment, skin disease treatment, promote cell regeneration and hair growth. A method of treating skin injury or skin disease in a patient comprising applying a composition comprising a volatile fraction from an ingredient comprising to a patient's wound, the volatile fraction comprising:
(A) feed the raw crude extract in about 2% alcohol; and (b) evaporate the crude extract at a pressure of less than about 1 atmosphere and a temperature of less than about 110 ° C. to produce a volatile fraction. obtain;
A method prepared by a process comprising steps.
（ａ）約２％のアルコール中に原料の粗抽出物を供給し；そして
（ｂ）約１気圧未満の気圧及び約１１０℃未満の温度で、粗抽出物を蒸発させて揮発性画分を得る；
ステップを含むプロセスによって調製される方法。 A sufficient amount of about 80% soybean seed, about 10% sesame seed and about 10% effective to promote skin injury treatment, skin disease treatment, enhanced cell regeneration and hair growth. A method of treating skin injury or skin disease in a patient, comprising applying to the patient's wound a composition comprising a volatile fraction from a raw material comprising a dried rhizome of palm leaf yellow or rhubarb, the volatile The fraction is:
(A) feed the raw crude extract in about 2% alcohol; and (b) evaporate the crude extract at a pressure of less than about 1 atmosphere and a temperature of less than about 110 ° C. to produce a volatile fraction. obtain;
A method prepared by a process comprising steps.
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| US11452753B2 (en) * | 2016-06-06 | 2022-09-27 | Charsire Biotechnology Corp. | Soybean seed extract, method for producing the same and uses thereof |
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| CN106442785A (en) * | 2016-09-27 | 2017-02-22 | 华润三九医药股份有限公司 | Establishment method of fingerprint spectrum of dayuan decoction compound and fingerprint spectrum |
| CN106420490A (en) * | 2016-11-18 | 2017-02-22 | 广州环亚化妆品科技有限公司 | Traditional Chinese medicine composition with effect of removing acne and preparation method and application of traditional Chinese medicine composition |
| CN109528506A (en) * | 2018-12-14 | 2019-03-29 | 安徽大学 | A kind of multiple-effect blemish clearing gel |
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