TWI520741B - 含有鮭魚魚精萃取物、啤酒酵母萃取物、大麥嫩葉萃取物及雞軟骨膠原蛋白之組合物用於製造口服用脂肪分解促進劑的用途 - Google Patents
含有鮭魚魚精萃取物、啤酒酵母萃取物、大麥嫩葉萃取物及雞軟骨膠原蛋白之組合物用於製造口服用脂肪分解促進劑的用途 Download PDFInfo
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- Physiology (AREA)
Description
本發明係關於一種具有脂肪分解促進作用且其胃蛋白酶及胰蛋白酶分解物具有血管收縮素轉化酶(ACE,Angiotensin Converting Enzyme)抑制作用之組合物,更詳細而言,本發明係關於一種包含鮭魚魚精萃取物、啤酒酵母萃取物、大麥嫩葉萃取物及雞軟骨膠原蛋白(Chicken Collagen)之組合物,含有上述組合物之口服用之脂肪分解促進劑及口服用之ACE抑制劑。
脂肪係以食物形式攝取之剩餘能量堆積於白色脂肪組織而成者,過剩之白色脂肪組織之堆積會導致肥胖,其結果,不僅會引起各種生活習慣疾病,在美容方面亦成為較大問題。尤其是最近科學上已明確,形成於內臟周圍(腸間膜脂肪)之白色脂肪組織之增大會誘發高血壓、胰島素阻抗、葡萄糖失耐、高脂血症,由此引起新陳代謝症候群,社會上迫切期望其預防及改善。
若脂肪細胞中堆積脂肪而肥大,則來自脂肪細胞之良性脂肪素之分泌減少,而分泌各種惡性脂肪素(例如TNFα(tumor necrosis factor α,腫瘤壞死因子α)、抗胰島素激素(resistin)等),引起胰島素阻抗(即胰島素敏感性降低)。其結果,無法充分降低血糖值,故為控制血糖值而分泌過剩之胰島素,導致高胰島素血症。若形成高胰島素血症,則由於過剩之胰島素對脂質代謝等之作用而引起新陳代謝症候群。
脂聯素(adiponectin)係良性脂肪素之一,其促進脂肪酸之燃燒及糖之攝取,改善胰島素阻抗(非專利文獻1)。脂聯素引起之脂肪酸之燃燒作用並非於脂肪細胞中產生,而係由肝臟及骨骼肌中之AMP(adenosine monophosphate,腺苷酸)活化蛋白激酶(AMPK,AMP-activated protein kinase)之活化所引起。於肝臟中,抑制糖之新生而燃燒脂肪酸,而且於骨骼肌中,攝取糖而使脂肪酸燃燒。
脂聯素之表現係隨著由前驅脂肪細胞向脂肪細胞之分化而被誘導。而且,脂肪細胞未肥大時係自脂肪細胞活躍地分泌。另一方面,於脂肪細胞已肥大之情形時,由於上述TNFα等而脂聯素之作用減弱。又,若脂肪細胞肥大,則脂聯素之轉錄受到抑制,脂聯素缺乏而引起代謝異常(非專利文獻1)。
有報告指出,來自蔬菜或果實之類胡蘿蔔素抑制胰島素誘導時之由前驅脂肪細胞向脂肪細胞之分化(專利文獻1),但如上所述,若抑制由前驅脂肪細胞向脂肪細胞之分化,則脂聯素之表現亦受到抑制,因此懷疑抑制向脂肪細胞之分化是否直接與抗肥胖作用有關。
先前以來,關於肥胖之預防及改善,已進行了節食、過剩能量之吸收之延遲及抑制、或消化管中之抑制糖質吸收之物質之探索等限制攝取能量之類的各種研究。然而,能量攝取之限制有時亦成為降低基礎代謝量之主要原因,未必改善肥胖。因此,為解決肥胖,較理想的是將已堆積之脂肪積極地代謝、分解並以熱能之形式發散。由此,近年來,自食品素材中活躍地進行具有脂肪分解促進作用之功能性成分之探索,提出了大量之脂肪分解促進劑及飲食品。
作為已知之源自天然之脂肪分解促進劑之有效成分,有芸香科植物(專利文獻2),薊族植物(專利文獻3),胡椒科植物(專利文獻4),橙葉、橙花、款冬葉及菖蒲根(專利文獻5),薏苡、大麥、決明子、蕃石榴及普洱茶(專利文獻6)等。進而,最近可列舉:將蓮或其萃取物作為有效成分之脂肪分解促進劑(專利文獻7)、含有樺木科白樺之萃取液及禾本科熊笹草之萃取物之至少一者之脂肪分解促進劑(專利文獻8)、含有小麥蛋白質之水解物之脂肪堆積抑制及促進劑(專利文獻9)等。
另一方面,高血壓係具代表性的新陳代謝症候群之症狀,其患者數逐年增加。已知高血壓會引起腦出血、蛛網膜下出血、腦梗塞、心肌梗塞、心絞痛、腎硬化症等各種併發症,對高血壓之發病機制已進行了各種研究,已知參與血壓升高之腎素-血管緊縮素系統(renin-angiotensin system)及參與降血壓之血管舒緩素-激肽系統(kallikrein-kinin system)發揮重要作用。腎素-血管緊縮素系統中,主要由肝臟分泌之血管緊縮素原(Angiotensinogen)藉由腎臟中所產生之腎素而形成血管緊縮素I,進而藉由血管收縮素轉化酶(ACE)而轉化為血管緊縮素II。該血管緊縮素II使血管平滑肌收縮、使血壓上升。另一方面,降血壓系統之血管舒緩素作用於激肽原而產生緩激肽。該緩激肽有擴張血管降低血壓之效果,而ACE有分解該緩激肽之作用。如此表明,ACE藉由作為升壓肽之血管緊縮素II之產生及作為降壓肽之緩激肽之失活的2種作用而參與血壓之上升。因此,可藉由抑制該ACE之酵素活性而抑制血壓之上升。作為ACE抑制活性物質而開發出之脯胺酸衍生物即卡托普利(captopril)或依那普利(enalapril)等被廣泛用於治療高血壓。
又,最近報告,作為食品素材蛋白質之酵素分解物之肽具有ACE抑制活性。例如大量報告有:明膠之膠原酶分解物(專利文獻10)、酪蛋白之胰蛋白酶分解物(專利文獻11~16)、γ-玉米醇溶蛋白(γ-zein)之嗜熱菌蛋白酶分解物(專利文獻17)、沙丁魚肌肉之胃蛋白酶分解物(專利文獻18)、柴魚之嗜熱菌蛋白酶分解物(專利文獻19)、芝麻蛋白之嗜熱菌蛋白酶分解物(專利文獻20)、κ-酪蛋白之胃蛋白酶等之分解物(專利文獻21)等。
專利文獻1:日本專利特開2003-95930號公報
專利文獻2:日本專利特開平8-81382號公報
專利文獻3:日本專利特開平8-301780號公報
專利文獻4:日本專利特開平8-245410號公報
專利文獻5:日本專利特開平11-228431號公報
專利文獻6:日本專利特開2002-275078號公報
專利文獻7:日本專利特開2004-307365號公報
專利文獻8:日本專利特開2006-045120號公報
專利文獻9:日本專利特開2007-106683號公報
專利文獻10:日本專利特開昭52-148631號公報
專利文獻11:日本專利特開昭58-109425號公報
專利文獻12:日本專利特開昭59-44323號公報
專利文獻13:日本專利特開昭60-23086號公報
專利文獻14:日本專利特開昭60-23087號公報
專利文獻15:日本專利特開昭61-36226號公報
專利文獻16:日本專利特開昭61-36227號公報
專利文獻17:日本專利特開平2-32127號公報
專利文獻18:日本專利特開平3-11097號公報
專利文獻19:日本專利特開平4-144696號公報
專利文獻20:日本專利特開平8-231588號公報
專利文獻21:日本專利特開平8-269088號公報
非專利文獻1:門脅孝等人,「脂聯素與糖尿病‧心血管病之分子機制」,第128次日本醫學會研討會記錄集「糖尿病與動脈硬化」2004年12月2日,34-45頁
本發明之目的在於提供一種具有脂肪分解促進作用及ACE抑制作用、且作為食品在副作用或毒性等安全性方面不存在問題、可口服之組合物。
本發明者對各種食材就其脂肪分解促進作用及ACE抑制活性進行了銳意研究,結果發現,將鮭魚魚精萃取物、啤酒酵母萃取物、大麥嫩葉萃取物及雞軟骨膠原蛋白(Chicken Collagen)組合製備之混合物具有優異之脂肪分解促進作用,進而,使用作為消化酵素之胃蛋白酶及胰蛋白酶對大麥嫩葉、鮭魚魚精萃取物、啤酒酵母萃取物、雞軟骨膠原蛋白之混合物進行處理之消化混合物表現出ACE抑制活性,從而完成了本發明。
即,本發明係關於:
(1)一種組合物,其特徵在於:其係含有鮭魚魚精萃取物、啤酒酵母萃取物、大麥嫩葉萃取物及雞軟骨膠原蛋白者,具有脂肪分解促進作用,且其胃蛋白酶及胰蛋白酶分解物具有血管收縮素轉化酶(ACE)抑制作用;
(2)如上述(1)之組合物,其中相對於大麥嫩葉萃取物100重量份,含有鮭魚魚精萃取物14.0~14.5重量份、啤酒酵母萃取物2.8~3.2重量份及雞軟骨膠原蛋白5.7~6.2重量份;
(3)如上述(1)或(2)之組合物,其中鮭魚魚精萃取物係藉由對鮭魚魚精進行酵素處理而獲得之包含分解成寡核苷酸(oligonucleotide)及寡肽之低分子成分之萃取物;
(4)如上述(1)或(2)之組合物,其中啤酒酵母萃取物係藉由對啤酒酵母進行酵素處理而獲得之含有低分子化之RNA(Ribonucleic Acid,核糖核酸)成分之萃取物;
(5)如上述(1)或(2)之組合物,其中大麥嫩葉萃取物含有30%之難消化性糊精;
(6)一種口服用脂肪分解促進劑,其特徵在於含有如上述(1)至(5)中任一項之組合物;
(7)一種口服用血管收縮素轉化酶(ACE)抑制劑,其特徵在於含有如上述(1)至(5)中任一項之組合物;以及
(8)一種飲食品,其特徵在於含有如上述(1)至(5)中任一項之組合物。
根據本發明,可提供一種具有優異之脂肪分解促進作用及其消化物具有ACE抑制作用之組合物。本發明之組合物由於作為食品不存在副作用或毒性等問題,故可長時間連續地簡便攝取,對肥胖或高血壓等成人病預防‧改善極為有用。
作為本發明之具有脂肪分解促進作用且藉由胃蛋白酶及胰蛋白酶進行分解後具有血管收縮素轉化酶(ACE)抑制作用之組合物,只要為含有鮭魚魚精萃取物、啤酒酵母萃取物、大麥嫩葉萃取物及雞軟骨膠原蛋白(Chicken Collagen)者則並無特別限制,特別好的是相對於大麥嫩葉萃取物100重量份而含有鮭魚魚精萃取物14.0~14.5重量份、啤酒酵母萃取物2.8~3.2重量份及雞軟骨膠原蛋白5.7~6.2重量份之組合物。
作為上述鮭魚魚精萃取物,只要為可作為食品用而使用者則並無特別限制,較好的是藉由對鮭魚魚精進行酵素處理而含有分解成寡核苷酸及寡肽之低分子成分之鮭魚魚精萃取物。特別好的是含有20~50%之分子量為1000~3000之寡核苷酸、寡肽之鮭魚魚精萃取物。此種鮭魚魚精萃取物例如可自日生生物股份有限公司等獲取。
作為上述啤酒酵母萃取物,只要為可作為食品用而使用者則並無特別限制,較好的是藉由對啤酒酵母進行酵素處理而含有低分子化之RNA之啤酒酵母萃取物。特別好的是含有20~50%之分子量為1000~3000之RNA之啤酒酵母萃取物。此種啤酒酵母萃取物例如可自日生生物股份有限公司等獲取。
作為上述大麥嫩葉萃取物,並無特別限制,可合適地使用食品用中於低溫下經噴霧乾燥之大麥嫩葉萃取物。亦可使用於大麥嫩葉萃取物中含有難消化性糊精者。作為含有難消化性糊精之大麥嫩葉萃取物,可合適地列舉含有20~30%之難消化性糊精之Luxon公司製造之大麥嫩葉萃取物。
作為上述雞軟骨膠原蛋白並無特別限制,可合適地使用食品用之雞軟骨膠原蛋白,例如可較佳地列舉LS Factory股份有限公司製造之雞軟骨膠原蛋白。
本發明之組合物除了口服用之脂肪分解促進劑使用以外,亦可用作口服用之脂肪堆積抑制劑、肥胖預防劑、肥胖改善劑,又,除口服用ACE抑制劑之外,可用作與卡托普利等合成醫藥品不同、攝取後無副作用或副作用極少之血壓上升抑制劑、高血壓之預防或改善劑。進而,藉由將本發明之組合物調配至飲食品中使用,可對飲食品賦予脂肪分解促進作用及ACE抑制作用。如下述實施例中所記載般,本發明之組合物於經作為消化酵素之胃蛋白酶及胰蛋白酶消化之後,表現出優異之ACE抑制作用,因此可藉由以食品之形式口服而於高血壓之預防‧改善方面發揮效果。為將本發明之組合物調配至飲食品中使用,將其有效成分之有效量於飲食品之製造原料階段或製造出之產品之階段等中添加、調配。此處,所謂「有效成分之有效量」,係指將各飲食品攝取通常食用之量之情形時攝取有效成分之含量,可依據接受者之年齡及體重、症狀、投予時間、劑形、投予方法、藥劑之組合等決定。
本發明之飲食品只要為含有本發明之組合物者則並無特別限制,具體可列舉以下者為例:將本發明之組合物直接製備成飲食品或補充品者,進而調配有各種蛋白質、糖類、脂肪、微量元素、維生素類等者,製成液狀、半液體狀或固體狀者,或添加、調配至普通之飲食品中而成者。
進而,本發明之飲食品亦可為健康食品、功能性食品、特定保健用食品或病人用食品。關於本發明之口服用脂肪分解促進劑、口服用ACE抑制劑及飲食品之製造,可使用通常所用之添加物或調配劑,例如賦形劑、增量劑、黏合劑、崩解劑、潤滑劑、分散劑、保存劑、濕潤劑、溶解助劑、防腐劑、穩定材、膠囊基劑、食品素材等助劑。作為該助劑之成分,例如可具體列舉:乳糖、果糖、葡萄糖、澱粉、明膠、碳酸鎂、合成矽酸鎂、滑石、硬脂酸鎂、碳酸鈣、甲基纖維素、羧甲基纖維素或其鹽、阿拉伯膠、聚乙二醇、糖漿、凡士林、甘油、乙醇、丙二醇、檸檬酸、氯化鈉、亞硫酸鈉、磷酸鈉、普魯蘭多糖(pullulan)、鹿角菜膠(carrageenan)、糊精、還原性異麥芽酮糖醇、山梨糖醇、木糖醇、甜菊甙、合成甜味劑、檸檬酸、抗壞血酸、酸味劑、碳酸氫鈉、蔗糖酯、植物氫化油脂、氯化鉀、紅花油、蜂蠟、大豆卵磷脂、香料等。
以下,根據實施例對本發明加以更具體說明,但本發明之技術範圍不限定於該等例示。
於本發明之實施例中,評價中所用之試樣係將鮭魚魚精萃取物(日生生物股份有限公司製造)、啤酒酵母萃取物(日生生物股份有限公司製造)、大麥嫩葉萃取物(Luxon公司製造,含有30%之難消化性糊精)、雞軟骨膠原蛋白(Chicken Collagen,LS Factory股份有限公司製造)單獨或組合製備。將用於製備試樣之各成分之濃度示於表1。
將小鼠前驅脂肪細胞株3T3-L1細胞於96well培養板上以成為3×104 cells/250 μl/well之方式播種,使用含有10% CS(calf serum,小牛血清)之DMEM(Dulbecco's Modified Eagle's Medium,達爾伯克氏改良伊格爾培養基)培養基(10% CS/DMEM)預培養2天,培養至變融合(day0)為止。將脂肪細胞分化誘導劑[0.5 mM異丁基甲基黃嘌呤(IBMX,isobutyl-methylxanthine)、1 μM地塞米松(DEX,Dexamethasone)、10 μg/ml胰島素]添加至培養基中,進而培養3天,藉此進行向脂肪細胞之分化誘導。將分化誘導後之細胞於含有10 μg/ml胰島素之10%FBS(Fetal Bovine Serum,胎牛血清)/DMEM培養基中培養3天之後,於基本培養基中培養6天。於該6天之培養中,每隔2天更換培養基使脂肪滴堆積。其後,單獨添加表1所記載之混合物及鮭魚魚精萃取物、啤酒酵母萃取物、大麥嫩葉萃取物、雞軟骨膠原蛋白,進而進行48小時之培養。培養結束後回收上清液,使用Adipolysis assay kit測定培養上清液中之甘油量。將該培養上清液中之甘油量作為脂肪分解之指標。甘油游離率係將對照值設為100%之情形時之相對值。
脂肪分解促進率%=[A/B]×100
A:添加萃取物時之游離甘油量
B:未添加萃取物時之游離甘油量
如上所述,脂肪分解促進作用之判定係將藉由脂肪分解而生成之甘油量之測定值作為指標求出。將經圖表化之結果示於圖1,將經數值化之結果示於圖2。於添加有含有大麥嫩葉萃取物之混合物(試樣)之區中,於1 mg/ml及5 mg/ml濃度下,分別顯示control(對照值)之208.67%、504.17%之值。另一方面,關於單獨添加各成分之區中之脂肪分解率,於大麥萃取物810 μg/ml添加區中顯示141.57%,於4.05 mg/ml添加區中顯示255.22%;於鮭魚魚精萃取物116 μg/ml添加區中顯示95%,於580 μg/ml添加區中顯示95.44%;於啤酒酵母萃取物24 μg/ml添加區中顯示95%,於120 μg/ml添加區中顯示81.6%;於雞軟骨膠原蛋白48 μg/ml添加區中顯示87.67%,於240 μg/ml添加區中顯示116.7%。如上所述表明,將大麥嫩葉萃取物、鮭魚魚精萃取物、啤酒酵母萃取物、雞軟骨膠原蛋白成分組合製備之試樣所表現出之顯著之脂肪分解促進作用於單獨添加各成分時未表現出,且顯示,藉由將大麥嫩葉萃取物、鮭魚魚精萃取物、啤酒酵母萃取物、雞軟骨膠原蛋白成分組合,可表現出顯著之協同效果。
進而,使用Cell counting Kit-8(Dojindo公司製造)對回收上清液之後之細胞生存率進行測定。如圖2所示之結果般,可確認混合物及各成分之添加區中生存率未見差異。
使用0.1M HCl‧KCl(pH值2.5)製備表1中所示之混合物及各成分之10%溶液,添加胃蛋白酶,於37℃下酵素處理2.5小時。處理後,於90℃沸騰水浴中加熱10分鐘,使胃蛋白酶失活。使用0.1M磷酸鉀緩衝液(pH值7.5)對該處理液進行5倍稀釋,進而添加胰蛋白酶,於37℃下酵素處理24小時。處理後,於90℃沸騰水浴中加熱10分鐘,使胰蛋白酶失活。對該處理液進行離心,冷凍乾燥上清液,用於以下之實驗。
ACE抑制活性之測定係根據Lieberman測定法之變形方法進行。將如上述般經酵素處理之冷凍乾燥試樣以與酵素處理前之濃度同樣地達到表1中所記載之濃度之方式進行製備。於該製備液(30 μL)中添加以含有400 mM NaCl之硼酸緩衝液(pH值8.3)稀釋至5 mM之Hip-His-Leu(Hippuryl-histidyl-leucine,馬尿酸-組胺酸-亮胺酸)基質溶液(250 μl),於37℃之恆溫水槽中保溫5分鐘。添加600 mu/ml(6 mU)之ACE溶液(100 μl),立即攪拌後,於37℃下反應60分鐘。添加1 N鹽酸(250 μl),攪拌而使反應停止之後,添加乙酸乙酯(1.5 ml)並充分攪拌,藉此使馬尿酸游離。於3,000 rpm、10分鐘之離心分離後回收上層之乙酸乙酯層1.0 ml,利用蒸發器蒸乾。進而於減壓乾燥器內使其乾燥60分鐘後,添加3 ml超純水,測定該溶液之吸光值(228 nm)。再者,抑制活性係根據下式作為抑制率(%)而算出:
抑制率(%)={(EC-ES)/(EC-EB)}×100
此處之ES係表示添加試樣溶液時之吸光值,EC係表示代替試樣溶液而添加超純水時之吸光值,EB係表示預先添加1 N鹽酸使其反應時之吸光值。
如圖3所示之結果表明,混合物1 mg/ml、10 mg/ml之情形與對應濃度之各組合物單獨之情形相比,ACE抑制活性明顯較高,且可見協同效果。
製備棒型之補充品,其係於每一支中調配有大麥嫩葉萃取物1687 mg、鮭魚魚精萃取物240 mg、啤酒酵母萃取物50 mg、雞軟骨膠原蛋白100 mg、難消化性糊精723 mg、糊精500 mg。
圖1係表示於本發明之組合物之存在下培養的脂肪細胞之培養上清液中的甘油量與於鮭魚魚精萃取物、啤酒酵母萃取物、大麥嫩葉萃取物或雞軟骨膠原蛋白之單獨存在下培養的脂肪細胞之培養上清液中的甘油量相比顯著增加的圖。圖之縱軸表示將無添加區之培養上清液中的甘油量作為基準之百分率之值;
圖2係表示於本發明之組合物之存在下培養的脂肪細胞之生存率與於鮭魚魚精萃取物、啤酒酵母萃取物、大麥嫩葉萃取物或雞軟骨膠原蛋白之單獨存在下培養的脂肪細胞生存率之間未見顯著差異的圖。圖之縱軸表示將無添加區之脂肪細胞之生存率作為基準之百分率之值;及
圖3係表示本發明之組合物之胃蛋白酶及胰蛋白酶處理物之ACE抑制活性與經胃蛋白酶及胰蛋白酶處理之鮭魚魚精萃取物、啤酒酵母萃取物、大麥嫩葉萃取物或雞軟骨膠原蛋白處理物單獨之ACE抑制活性相比明顯優異之圖。
(無元件符號說明)
Claims (4)
- 一種含有鮭魚魚精萃取物、啤酒酵母萃取物、大麥嫩葉萃取物及雞軟骨膠原蛋白之組合物的用途,於該組合物中,相對於大麥嫩葉萃取物100重量份,含有鮭魚魚精萃取物14.0~14.5重量份、啤酒酵母萃取物2.8~3.2重量份及雞軟骨膠原蛋白5.7~6.2重量份,該用途之特徵在於:其係用於製造口服用脂肪分解促進劑者;上述組合物具有脂肪分解促進作用,且其胃蛋白酶及胰蛋白酶分解物具有血管收縮素轉化酶(ACE)抑制作用。
- 如請求項1之用途,其中鮭魚魚精萃取物係藉由對鮭魚魚精進行酵素處理而獲得之含有分解成寡核苷酸及寡肽之低分子成分之萃取物。
- 如請求項1之用途,其中啤酒酵母萃取物係藉由對啤酒酵母進行酵素處理而獲得之含有低分子化之RNA成分之萃取物,上述RNA之分子量為1000~3000。
- 如請求項1之用途,其中大麥嫩葉萃取物含有30%之難消化性糊精。
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