TW201701890A - Yeast extract having vasorelaxing effect - Google Patents
Yeast extract having vasorelaxing effect Download PDFInfo
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- TW201701890A TW201701890A TW105116361A TW105116361A TW201701890A TW 201701890 A TW201701890 A TW 201701890A TW 105116361 A TW105116361 A TW 105116361A TW 105116361 A TW105116361 A TW 105116361A TW 201701890 A TW201701890 A TW 201701890A
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- yeast extract
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- yeast
- glutathione
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- 239000012138 yeast extract Substances 0.000 title claims abstract description 49
- 229940041514 candida albicans extract Drugs 0.000 title claims abstract description 48
- 230000001196 vasorelaxation Effects 0.000 title abstract 2
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims abstract description 19
- 230000002883 vasorelaxation effect Effects 0.000 claims abstract description 7
- 239000004480 active ingredient Substances 0.000 claims abstract description 6
- RWSXRVCMGQZWBV-WDSKDSINSA-N glutathione Chemical compound OC(=O)[C@@H](N)CCC(=O)N[C@@H](CS)C(=O)NCC(O)=O RWSXRVCMGQZWBV-WDSKDSINSA-N 0.000 claims description 44
- OIRDTQYFTABQOQ-KQYNXXCUSA-N adenosine Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](CO)[C@@H](O)[C@H]1O OIRDTQYFTABQOQ-KQYNXXCUSA-N 0.000 claims description 28
- 108010024636 Glutathione Proteins 0.000 claims description 23
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Classifications
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- A—HUMAN NECESSITIES
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- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
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- A61K31/33—Heterocyclic compounds
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- A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
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- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7042—Compounds having saccharide radicals and heterocyclic rings
- A61K31/7052—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides
- A61K31/706—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom
- A61K31/7064—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom containing condensed or non-condensed pyrimidines
- A61K31/7076—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom containing condensed or non-condensed pyrimidines containing purines, e.g. adenosine, adenylic acid
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/06—Fungi, e.g. yeasts
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- A—HUMAN NECESSITIES
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- A61K38/00—Medicinal preparations containing peptides
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- A—HUMAN NECESSITIES
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- A61K38/04—Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
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- A—HUMAN NECESSITIES
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- A—HUMAN NECESSITIES
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- A61P9/00—Drugs for disorders of the cardiovascular system
- A61P9/12—Antihypertensives
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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Abstract
Description
本發明提供含有來自圓酵母之水性溶劑萃取物作為有效成分，且血管鬆弛作用優良之食品組成物及醫藥組成物。The present invention provides a food composition and a pharmaceutical composition comprising an aqueous solvent extract derived from round yeast as an active ingredient and having excellent vascular relaxation effects.
高血壓症因為缺乏自覺症狀，會在未察覺時惡化，有時也會對其他臟器造成不好的影響，作為併發症併發各種血管性疾病。就主要的併發症而言可列舉心絞痛、心肌梗塞、腦梗塞、腎功能低落、剝離性主動脈瘤等。此等中，也有若惡化嚴重時會致命者。Because of the lack of self-conscious symptoms, hypertension can worsen when it is not noticed, and sometimes it can cause adverse effects on other organs, as a complication and various vascular diseases. Examples of major complications include angina pectoris, myocardial infarction, cerebral infarction, renal dysfunction, and dissected aortic aneurysm. In this case, there are also those who are fatal if they deteriorate seriously.
就高血壓症之治療方法而言，有生活習慣的改善及藥物治療。就生活習慣的改善一般而言可列舉飲食控制、減量、酒精的控制、適度的運動、禁菸等。此等係因為不使用藥物而有副作用的隱憂少或經濟上負擔少的優點，但如食物鹽分限制之類的飲食控制或減量係需要長時間才能使症狀有所改善，有時會成為患者的負擔。As far as the treatment of hypertension is concerned, there are improvements in lifestyle and medication. The improvement of living habits generally includes diet control, reduction, alcohol control, moderate exercise, and smoking ban. These are advantages of less side effects or less economic burden because they do not use drugs, but diet control or reduction such as food salt restriction takes a long time to improve symptoms, sometimes it becomes a patient's burden.
另一方面，就藥物療法而言，係依據對象之血壓高低或併發症的有無來選擇適當之藥劑，可列舉利尿劑、血管張力素轉化酶抑制劑、鈣拮抗劑、交感神經阻斷藥等。此等醫藥品主要為醫療機關中於重症之高血壓患者上所使用者，雖然有效性高，但必須長期持續服用，且不能夠自行判斷而去服用或中止服用。另外，也有著副作用多的課題存在。On the other hand, in the case of drug therapy, an appropriate agent is selected depending on the blood pressure level of the subject or the presence or absence of a complication, and examples thereof include diuretics, angiotensin converting enzyme inhibitors, calcium antagonists, sympatholytic inhibitors, and the like. . These medicines are mainly used by patients with severe hypertension in medical institutions. Although they are highly effective, they must be taken continuously for a long time and cannot be taken or suspended at their own discretion. In addition, there are also problems with many side effects.
因此，為了高血壓患者、高血壓危險群之高血壓症的預防、改善，而有副作用少、安全且可持續地實施之方法的需求，到目前為止已有許多的健康食品開發出來。例如已知有裙帶菜肽(專利文獻1)、杜仲葉配糖體(專利文獻2)、鰹魚乾肽(專利文獻3)、乳三肽(專利文獻4)、沙丁魚肽(專利文獻5)、γ-胺基丁酸摻合食品等，但鮮少有人知道酵母萃取物具有血管鬆弛作用。 [先前技術文獻] [專利文獻]Therefore, in order to prevent and improve hypertension caused by hypertensive patients and high blood pressure risk groups, there has been a demand for a method for implementing safe and sustainable methods with few side effects, and many health foods have been developed so far. For example, wakame peptide (Patent Document 1), Eucommia ulmoides (Patent Document 2), squid peptide (Patent Document 3), lactotripeptide (Patent Document 4), and sardine peptide (Patent Document 5) are known. Γ-aminobutyric acid is blended with foods, etc., but few people know that yeast extract has a vasoactive effect. [Prior Technical Literature] [Patent Literature]
[專利文獻1]日本特開2002-138100號公報 [專利文獻2]日本特開2008-106008號公報 [專利文獻3]日本特開2014-5243號公報 [專利文獻4]日本特開平11-98978號公報 [專利文獻5]日本特開2006-56805號公報[Patent Document 1] JP-A-2008-106008 [Patent Document 3] JP-A-2014-5243 [Patent Document 4] Japanese Patent Laid-Open No. 11-98978 Japanese Patent Publication No. 2006-56805
[發明所欲解決之課題] 本發明提供對於高血壓患者、高血壓危險群副作用少，安全且低廉，在食品領域或醫藥品領域中之有效用之來自食物的血管鬆弛劑。 [解決課題之手段][Problems to be Solved by the Invention] The present invention provides a vascular relaxing agent derived from food which is effective for use in a food field or a pharmaceutical field, and which has few side effects and is safe and inexpensive for hypertensive patients and high blood pressure risk groups. [Means for solving the problem]
本發明中，對於從大鼠摘出之胸部主動脈血管確認了血管鬆弛作用，發現了藉由將酵母進行熱水處理、酸或鹼處理、酵素處理之簡單的操作而獲得之酵母萃取物具有血管鬆弛作用。 [發明之效果]In the present invention, the vasorelaxation effect is confirmed for the thoracic aorta blood vessels taken from the rat, and the yeast extract obtained by the simple operation of hot water treatment, acid or alkali treatment, and enzyme treatment of the yeast is found to have blood vessels. Relaxation effect. [Effects of the Invention]
本發明之血管鬆弛劑因為是從已知為食品的酵母所萃取而得之萃取物，不僅生產較容易且價格低廉，而且高血壓患者、高血壓危險群為了高血壓症的預防、改善，可安全且長期持續地將本發明之血管鬆弛劑作為食品來攝取。Since the vascular relaxant of the present invention is an extract obtained by extracting from a yeast known as a food, it is not only easy to produce but also inexpensive, and the hypertensive patient and the high blood pressure risk group can prevent and improve hypertension. The vascular relaxing agent of the present invention is ingested as a food safely and continuously for a long period of time.
製造本發明之酵母萃取物所使用之酵母只要是一般使用之酵母即可使用。具體而言可列舉麵包酵母、啤酒酵母、圓酵母等。其中，尤其期望為圓酵母(Candida utilis)。 本發明中使用之酵母萃取物係麩胱甘肽含量為15質量%以上、腺嘌呤含量為0.05質量%以上、腺苷含量為0.05質量%以上、甲基硫代腺苷含量為0.1質量%以上之酵母萃取物。更期望為麩胱甘肽含量為15質量%以上、腺嘌呤含量為0.1質量%以上、腺苷含量為0.1質量%以上、甲基硫代腺苷含量為0.2質量%以上之酵母萃取物。The yeast used for producing the yeast extract of the present invention can be used as long as it is a yeast which is generally used. Specific examples include baker's yeast, brewer's yeast, and round yeast. Among them, it is particularly desirable to be a Candida utilis. The yeast extract used in the present invention has a glutathione content of 15% by mass or more, an adenine content of 0.05% by mass or more, an adenosine content of 0.05% by mass or more, and a methylthio adenosine content of 0.1% by mass or more. Yeast extract. A yeast extract having a glutathione content of 15% by mass or more, an adenine content of 0.1% by mass or more, an adenosine content of 0.1% by mass or more, and a methylthio adenosine content of 0.2% by mass or more is more desirable.
如此之酵母萃取物係藉由從含有麩胱甘肽之酵母菌體萃取出酵母萃取物，而獲得麩胱甘肽含量為15質量%以上之酵母萃取物。酵母之培養形式雖然沒有特別之限制，一般而言可使用批式培養或連續培養之任一者。培養基也可使用一般所使用者。例如，就碳源而言使用葡萄糖、乙酸、乙醇、甘油、糖蜜、亞硫酸紙漿廢液等，就氮源而言使用尿素、氨、硫酸銨、氯化銨、硝酸鹽等。就磷酸、鉀、鎂源而言例如可使用過磷酸鈣、磷酸銨、氯化鉀、氫氧化鉀、硫酸鎂、氯化鎂等，其他還可添加鋅、銅、錳、鐵離子等無機鹽。除此之外，還可添加維生素、胺基酸、核酸相關物質等，或可添加酪蛋白、酵母萃取物、肉精、蛋白腖等有機物。培養溫度係21~37℃，宜為25~34℃，pH係3.0~8.0，尤其宜為3.5~7.0。Such a yeast extract is obtained by extracting a yeast extract from a yeast cell containing glutathione to obtain a yeast extract having a glutathione content of 15% by mass or more. The culture form of the yeast is not particularly limited, and generally, either batch culture or continuous culture can be used. The medium can also be used by a general user. For example, as the carbon source, glucose, acetic acid, ethanol, glycerin, molasses, sulfite pulp waste liquid or the like is used, and as the nitrogen source, urea, ammonia, ammonium sulfate, ammonium chloride, nitrate or the like is used. Examples of the phosphoric acid, potassium, and magnesium sources include superphosphate, ammonium phosphate, potassium chloride, potassium hydroxide, magnesium sulfate, and magnesium chloride. Other inorganic salts such as zinc, copper, manganese, and iron ions may be added. In addition, vitamins, amino acids, nucleic acid-related substances, and the like may be added, or organic substances such as casein, yeast extract, meat, and peptone may be added. The culture temperature is 21 to 37 ° C, preferably 25 to 34 ° C, and the pH is 3.0 to 8.0, particularly preferably 3.5 to 7.0.
在酵母菌體培養後進行本發明之酵母萃取物的萃取。酵母萃取物之萃取法係沒有特別之限制，一般而言，可為自消化法、熱水萃取法、酵素萃取法、酸或鹼萃取法，或是可藉由此等之組合進行萃取。Extraction of the yeast extract of the present invention is carried out after the yeast body culture. The extraction method of the yeast extract is not particularly limited, and generally, it may be a self-digestion method, a hot water extraction method, an enzyme extraction method, an acid or alkali extraction method, or may be extracted by a combination thereof.
藉由自消化來萃取酵母萃取物時，例如在55℃攪拌4小時。若是酵素萃取法，則以細胞壁溶解酵素或蛋白酶等進行攪拌萃取。若是酸萃取法，則以硫酸等調整至酸性後進行萃取。若是鹼萃取法，則調整至鹼性後進行萃取。或者，也可在自消化後，進行酵素萃取等組合此等方法來進行。When the yeast extract is extracted by self-digestion, for example, stirring at 55 ° C for 4 hours. In the case of the enzyme extraction method, the mixture is stirred and extracted by cell wall lysin or protease. In the case of the acid extraction method, the mixture is acidified with sulfuric acid or the like and then extracted. In the case of the alkali extraction method, the mixture is adjusted to be alkaline and then extracted. Alternatively, it may be carried out by a combination of enzyme extraction or the like after self-digestion.
萃取出酵母萃取物後藉由離心分離而分離出酵母殘渣，濃縮後藉由冷凍乾燥或熱風乾燥可獲得含有麩胱甘肽之酵母萃取物。酵母萃取物中所含有之麩胱甘肽係指還原型麩胱甘肽。After extracting the yeast extract, the yeast residue is separated by centrifugation, and after concentration, the yeast extract containing glutathione can be obtained by freeze-drying or hot air drying. The glutathione contained in the yeast extract means reduced glutathione.
本發明之酵母萃取物係酵母萃取物中麩胱甘肽含量為15質量%以上者。欲使酵母萃取物中麩胱甘肽含量為15%以上，盡可能地使用培養酵母中之麩胱甘肽含量高者，從該酵母菌體藉由上述之萃取法萃取出酵母萃取物則效率較良好。就使酵母中之麩胱甘肽含量提高的方法而言，可為公知方法。例如有在培養基中添加鋅離子之方法(日本特開2000-279164號公報)、以鎘耐性、巨環內酯系抗生素耐性作為指標之麩胱甘肽高含量酵母株的取得方法(日本特開2006-42637號公報、日本特開2006-42638號公報)。也可如日本特開2011-103789號公報，從變異酵母菌體進行萃取來獲得。此外，若為一般販售之酵母萃取物，例如有興人生命科學公司製之「HITHION™」YH-15。The yeast extract of the present invention has a glutathione content of 15% by mass or more in the yeast extract. If the content of glutathione in the yeast extract is 15% or more, and the content of glutathione in the cultured yeast is as high as possible, the efficiency of extracting the yeast extract from the yeast by the above extraction method is efficient. Better. A method for increasing the glutathione content in yeast can be a known method. For example, there is a method of adding zinc ions to a culture medium (JP-A-2000-279164), and a method for obtaining a high-content glutathione yeast strain using cadmium tolerance and macrolide antibiotic resistance as an index (Japanese Patent Co., Ltd.) Japanese Patent Publication No. 2006-42637 and Japanese Patent Laid-Open No. 2006-42638. It can also be obtained by extracting from a mutant yeast cell as disclosed in JP-A-2011-103789. In addition, if it is a yeast extract which is generally sold, for example, "HITHIONTM" YH-15 manufactured by Hiroshi Life Science Co., Ltd.
本案發明之麩胱甘肽濃度之測定方法係進行DTNB-HPLC法(Journal of Chromatography, 194(1980)424-428)。其中，本案之麩胱甘肽含量係指還原型麩胱甘肽含量。The method for measuring the concentration of glutathione in the present invention is subjected to DTNB-HPLC method (Journal of Chromatography, 194 (1980) 424-428). Among them, the glutathione content in the present case refers to the reduced glutathione content.
腺嘌呤、腺苷、甲基硫代腺苷濃度係可藉由下述之測定方法、條件進行定量。也就是說，將試料20mg以0.1w/w%甲酸混合定量至100mL，藉由液相層析串聯質譜儀分析法(LC-MS/MS)進行分析。將腺嘌呤、腺苷、甲基硫代腺苷(皆為自由態)之各試藥作為標準品，在相同條件下於LC-MS/MS進行測定，藉由該峰之面積製作檢量線，算出試料中之腺嘌呤、腺苷、甲基硫代腺苷之含量。 分析條件可如同以下設定。 ＜LC條件＞ ・管柱:Inertsil ODS-3(2.1mm×150mm) ・流動相:A:0.1w/w%甲酸、B:0.1w/w%甲酸/乙腈、A→B:20分鐘線性梯度 ・流量:0.2mL/分鐘 ・管柱溫度:45℃ ＜MS/MS條件＞ ・測定機器:amazon(Bruker daltonics公司) ・離子化方法:ESI-Positive ・離子監測:腺嘌呤:m/z136→136、腺苷:m/z268→136、甲基硫代腺苷:m/z298→136The concentration of adenine, adenosine, and methylthioadenosine can be quantified by the following measurement methods and conditions. That is, 20 mg of the sample was mixed with 0.1 w/w% formic acid to 100 mL, and analyzed by liquid chromatography tandem mass spectrometry (LC-MS/MS). Each of the reagents of adenine, adenosine, and methylthioadenosine (all free states) was used as a standard, and was subjected to LC-MS/MS under the same conditions, and a calibration curve was prepared by the area of the peak. The content of adenine, adenosine, and methylthioadenosine in the sample was calculated. The analysis conditions can be set as follows. <LC conditions> • Column: Inertsil ODS-3 (2.1 mm × 150 mm) ・ Mobile phase: A: 0.1 w/w% formic acid, B: 0.1 w/w% formic acid/acetonitrile, A→B: 20 minute linear gradient・Flow rate: 0.2 mL/min, column temperature: 45 °C <MS/MS conditions> ・Measuring equipment: amazon (Bruker daltonics) ・Ionization method: ESI-Positive ・Ion monitoring: Adenine: m/z 136→136 , adenosine: m/z268→136, methylthioadenosine: m/z298→136
本發明中，酵母萃取物係使用麩胱甘肽15質量%以上，且腺嘌呤含量為0.05質量%以上、腺苷含量為0.05質量%以上、甲基硫代腺苷含量為0.1質量%以上者。只要是該含量，使用之酵母萃取物並沒有限制，例如也可將兩種以上之酵母萃取物混合成為麩胱甘肽含量15質量%以上、腺嘌呤含量0.05質量%以上、腺苷含量0.05質量%以上、甲基硫代腺苷含量0.1質量%以上之酵母萃取物。更期望酵母萃取物為麩胱甘肽含量15質量%以上、腺嘌呤含量0.1質量%以上、腺苷含量0.1質量%以上、甲基硫代腺苷含量0.2質量%以上之酵母萃取物。In the present invention, the yeast extract is 15% by mass or more of glutathione, and the adenine content is 0.05% by mass or more, the adenosine content is 0.05% by mass or more, and the methylthio adenosine content is 0.1% by mass or more. . The yeast extract to be used is not limited as long as it is contained. For example, two or more yeast extracts may be mixed to have a glutathione content of 15% by mass or more, an adenine content of 0.05% by mass or more, and adenosine content of 0.05% by mass. A yeast extract having a % or more methyl thio adenosine content of 0.1% by mass or more. More preferably, the yeast extract is a yeast extract having a glutathione content of 15% by mass or more, an adenine content of 0.1% by mass or more, an adenosine content of 0.1% by mass or more, and a methylthio adenosine content of 0.2% by mass or more.
本發明之血管鬆弛劑之有效成分係酵母萃取物，故可直接攝取酵母萃取物作為血管鬆弛劑，此外也可製成與其他素材的混合物後攝取。Since the active ingredient of the vascular relaxant of the present invention is a yeast extract, the yeast extract can be directly ingested as a vascular relaxant, and a mixture with other materials can be used for ingestion.
本發明之血管鬆弛劑含有上述酵母萃取物作為有效成分，且指以食品、飲品、嗜好品、營養補充品、醫藥品等的方式經口攝取者。其形態沒有特別之限定。此外，就營養補充品或醫藥品之形態也沒有特別之限定，也可為錠劑、膠囊、軟膠囊、營養飲料狀之形態來攝取。The vascular relaxing agent of the present invention contains the above yeast extract as an active ingredient, and refers to a person who orally ingests foods, drinks, hobbies, nutritional supplements, pharmaceuticals, and the like. The form is not particularly limited. Further, the form of the nutritional supplement or the pharmaceutical is not particularly limited, and may be ingested in the form of a tablet, a capsule, a soft capsule, or a nutritious beverage.
在飲食品中上述組成物之摻合量係只要具有血管鬆弛作用便沒有特別之限定，例如本發明之酵母萃取物係相對於飲食品之重量100g含有10μg~20g即可。其中適宜為100μg~2g，更適宜為1mg~1g。The blending amount of the above-mentioned composition in the food or drink is not particularly limited as long as it has a vasoactive effect. For example, the yeast extract of the present invention may contain 10 μg to 20 g per 100 g of the weight of the food or beverage. Among them, it is preferably 100 μg to 2 g, more preferably 1 mg to 1 g.
醫藥品中組成物之摻合量沒有特別之限定，例如本發明之酵母萃取物係相對於組成物之重量100g含有10μg~20g即可。其中適宜為100μg~2g，更適宜為1mg~1g。 [實施例]The blending amount of the composition in the pharmaceutical product is not particularly limited. For example, the yeast extract of the present invention may contain 10 μg to 20 g per 100 g of the weight of the composition. Among them, it is preferably 100 μg to 2 g, more preferably 1 mg to 1 g. [Examples]
以下將列舉實施例來更詳細說明本發明，但本發明並非僅限定於此等實施例。The invention will be described in more detail below by way of examples, but the invention is not limited thereto.
(實施例1) 1.血管張力測定方法 a.試藥、材料 ・大鼠:雄性SD大鼠(8~11週) ・緩衝液:PSS(pH7.4)[NaCl 145、KCl 5、Na2 HPO4 1、CaCl2 2.5、MgSO4 ・7H2 O 0.5、葡萄糖 10、HEPES 5(mM)] ・收縮刺激劑:1.0μM脫羥腎上腺素(PE) b.製備 ・PE:溶解於PSS並使其成為10μM ・「HITHION™」YH-15:溶解於製備而得之10μM PE。使用之「HITHION™」YH-15係麩胱甘肽含量為19.0質量%、腺嘌呤含量為0.2質量%、腺苷含量為0.1質量%、甲基硫代腺苷含量為0.4質量%。(Example 1) 1. Method for measuring vascular tone a. Test drug, material, rat: male SD rat (8 to 11 weeks) • Buffer: PSS (pH 7.4) [NaCl 145, KCl 5, Na 2 HPO 4 1 , CaCl 2 2.5, MgSO 4 · 7H 2 O 0.5, glucose 10, HEPES 5 (mM)] • Contraction stimulating agent: 1.0 μM phenylephrine (PE) b. Preparation • PE: dissolved in PSS It became 10 μM ・"HITHIONTM" YH-15: 10 μM PE dissolved in the preparation. The "HITHIONTM" YH-15 system used had a glutathione content of 19.0% by mass, an adenine content of 0.2% by mass, an adenosine content of 0.1% by mass, and a methylthioadenosine content of 0.4% by mass.
c.測定流程 摘出大鼠胸部主動脈血管後，在37℃之PPS中，邊藉由混合氣體(95%O2 、5%CO2 )起泡邊進行45分鐘達到了平衡化。之後，除去脂肪組織並切斷為2~3mm後，作為血管環標本。於以4.5mL之PSS裝滿之器官浴槽(Organ Bath)中的掛鉤對上放置血管環標本，給予2g之負荷45分鐘使其平衡化。其中，平衡化中每15分鐘以經起泡處理之PSS更換器官浴槽中之溶液。平衡化後添加0.5mL之10μM PE(最終濃度:1μM)，使血管收縮。在收縮後之張力安定了以後，藉由將器官浴槽中之溶液以一定量逐次地等量置換為「HITHION™」YH-15溶液，開始進行張力變化之測定。結果如圖1所示。血管鬆弛率定義為令添加1μM PE後之張力增加量(ΔTc)為100時之添加「HITHION™」YH-15溶液後之張力減少量(ΔTs)的比率，藉由下式算出。 血管鬆弛率(%)=ΔTs/ΔTc×100 「HITHION™」YH-15之濃度與血管鬆弛率的關係表示於圖2。c. Measurement procedure After the rat thoracic aorta was removed, it was equilibrated in a PPS at 37 ° C for 45 minutes by bubbling with a mixed gas (95% O 2 , 5% CO 2 ). Thereafter, the adipose tissue was removed and cut into 2 to 3 mm, and then used as a blood vessel ring specimen. A vascular ring specimen was placed on the hook pair in an organ bath filled with 4.5 mL of PSS, and a load of 2 g was applied for 45 minutes to equilibrate it. Among them, the solution in the organ bath was replaced with the foamed PSS every 15 minutes in the equilibration. After equilibration, 0.5 mL of 10 μM PE (final concentration: 1 μM) was added to contract the blood vessels. After the tension after contraction was stabilized, the change in the tension was started by replacing the solution in the organ bath with a certain amount of the same amount of "HITHIONTM" YH-15 solution. The result is shown in Figure 1. The vascular relaxation rate is defined as the ratio of the amount of decrease in tension (ΔTs) after adding the "HITHIONTM" YH-15 solution when the amount of increase in tension (ΔTc) after adding 1 μM PE is 100, and is calculated by the following formula. Vascular relaxation rate (%) = ΔTs / ΔTc × 100 The relationship between the concentration of "HITHIONTM" YH-15 and the vascular relaxation rate is shown in Fig. 2 .
2.血管張力測定結果 確認了酵母萃取物「HITHION™」YH-15有濃度依存性的血管鬆弛作用 (圖1及圖2)。此外，獲得就抑制1.0μM PE之作用50%之濃度的EC50 值而言係7.16±5.48mg/mL。也就是說，可獲得具有血管鬆弛作用之酵母萃取物。而，針對作為陽性對照也同樣進行評價的血管鬆弛藥劑維拉帕米(Verapamil)，也確認有血管鬆弛效果(圖3及圖4)。EC50 值係1.66±0.74μg/mL。2. Vascular tension measurement results confirmed that the yeast extract "HITHIONTM" YH-15 has a concentration-dependent vasorelaxation effect (Fig. 1 and Fig. 2). Further, the EC 50 value obtained by suppressing the concentration of 50% of the action of 1.0 μM PE was 7.16 ± 5.48 mg / mL. That is, a yeast extract having a blood vessel relaxation effect can be obtained. On the other hand, Verapamil, a vasoactive agent which was also evaluated as a positive control, was also confirmed to have a vasoactive effect (Fig. 3 and Fig. 4). The EC 50 value was 1.66 ± 0.74 μg / mL.
(比較例1) 使用「NUCLEAMINE™」(興人生命科學(股)公司製)替代作為實施例1之酵母萃取物之「HITHION™」YH-15以外，與實施例1進行同樣之試驗。其中酵母萃取物「NUCLEAMINE™」係麩胱甘肽含量為0.8質量%、腺嘌呤含量為0.1質量%、腺苷含量為0.6質量%、甲基硫代腺苷含量為0.2質量%。對於「NUCLEAMINE™」所致之血管鬆弛作用進行評價，結果認定其不具有血管鬆弛作用。(Comparative Example 1) The same test as in Example 1 was carried out except that "HITHIONTM" YH-15, which is the yeast extract of Example 1, was replaced by "NUCLEAMINETM" (manufactured by Xingren Life Science Co., Ltd.). The yeast extract "NUCLEAMINETM" had a glutathione content of 0.8% by mass, an adenine content of 0.1% by mass, an adenosine content of 0.6% by mass, and a methylthioadenosine content of 0.2% by mass. The evaluation of the vasorelaxation effect by "NUCLEAMINETM" was carried out, and it was found that it did not have a vascular relaxation effect.
(比較例2) 使用「AROMILD™ G」(興人生命科學(股)公司製)替代作為實施例1之酵母萃取物之「HITHION™」YH-15以外，與實施例1進行同樣之試驗。其中，酵母萃取物「AROMILD™ G」係麩胱甘肽含量為0.1質量%、腺嘌呤含量為0.1%、腺苷含量為0.2%、未檢出有甲基硫代腺苷。對於「AROMILD™ G」所致之血管鬆弛作用進行評價，結果認定其不具有血管鬆弛作用。 [產業上利用性](Comparative Example 2) The same test as in Example 1 was carried out except that "HITHIONTM" YH-15, which is the yeast extract of Example 1, was replaced by "AROMILDTM G" (manufactured by Xingren Life Science Co., Ltd.). Among them, the yeast extract "AROMILDTM G" had a glutathione content of 0.1% by mass, an adenine content of 0.1%, an adenosine content of 0.2%, and no methylthioadenosine was detected. The evaluation of the vasorelaxation effect by "AROMILDTM G" was carried out, and it was found that it did not have a vascular relaxation effect. [Industrial use]
本發明之酵母萃取物具有主動脈鬆弛作用，除此之外本發明者們深入研究之結果得知，本酵母萃取物中被吸附於ODS管柱之分子量500以下的低分子級分具有強力之作用。血管平滑肌之緊張性與血管病性之疾病(例如血管萎縮、高血壓、動脈硬化等)或狀態的關係已為周知，從具有血管鬆弛作用之藥劑對於高血壓症、心臟衰竭、心絞痛、缺血性心臟病或心肌梗塞之治療及預防有效果，可以設想本發明之酵母萃取物也對於此等之疾病或狀態有效果。The yeast extract of the present invention has aortic relaxation effect, and in addition, as a result of intensive studies by the present inventors, it has been found that the low molecular fraction of the yeast extract which is adsorbed on the ODS column having a molecular weight of 500 or less is strong. effect. The relationship between vascular smooth muscle tone and vascular disease (such as vascular atrophy, hypertension, arteriosclerosis, etc.) or status is well known, from agents with vasorelaxic effects to hypertension, heart failure, angina pectoris, ischemia The treatment and prevention of sexual heart disease or myocardial infarction is effective, and it is conceivable that the yeast extract of the present invention is also effective for such diseases or conditions.
無no
[圖1]圖1係添加「HITHION™」YH-15時之血管張力層析圖。 [圖2]圖2係添加「HITHION™」YH-15時之血管鬆弛率(%)。 [圖3]圖3係添加維拉帕米(Verapamil)時之血管張力層析圖。 [圖4]圖4係表示添加維拉帕米時之血管鬆弛率(%)。[Fig. 1] Fig. 1 is a blood vessel tension chromatogram when "HITHIONTM" YH-15 was added. Fig. 2 is a graph showing the rate of vascular relaxation (%) when "HITHIONTM" YH-15 was added. [Fig. 3] Fig. 3 is a blood vessel tension chromatogram when Verapamil is added. Fig. 4 is a graph showing the rate of vascular relaxation (%) when verapamil was added.
無no
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