TWI448250B - Muscadine composition with improved anti-oxidant activity - Google Patents

Description

具有改良的抗氧化劑活性的麝香葡萄組成物Muscat composition with improved antioxidant activity

交互參照相關申請案Cross-reference related application

本申請案主張2008年7月31日申請之美國臨時申請案第61/085,369號及2008年7月31日申請之美國臨時申請案第61/085,371號的權益及優先權，該等申請案之每一者均以全文引用的方式併入本文中。The present application claims the benefit and priority of U.S. Provisional Application No. 61/085,369, filed on Jul. 31, 2008, and U.S. Provisional Application No. 61/085,371, filed on Jul. 31, 2008. Each is incorporated herein by reference in its entirety.

本揭示案係關於一種包括麝香葡萄果漿萃取物及白藜蘆醇之麝香葡萄組成物，其係用作抗氧化劑，諸如用作抗衰老補充劑。The present disclosure relates to a muscat grape composition comprising a muscadine pulp extract and resveratrol, which is used as an antioxidant, such as as an anti-aging supplement.

活性氧物質(reactive oxygen species，ROS)為需氧代謝之專性副產物且因此持續在人類及其他生物體內產生。人類亦接觸來自諸如污染、日光及飲食之外源性/環境來源之ROS。雖然存在ROS之不同化學形式，但其均對細胞組份及大分子之結構及功能產生有害作用。ROS產生/暴露之強度被稱為氧化應力(oxidative stress)。Reactive oxygen species (ROS) are obligate by-products of aerobic metabolism and thus continue to be produced in humans and other organisms. Humans also come into contact with ROS from sources/environmental sources such as pollution, sunlight and diet. Although there are different chemical forms of ROS, they all have harmful effects on the structure and function of cell components and macromolecules. The intensity of ROS production/exposure is called oxidative stress.

氧化應力與諸如糖尿病、癌症、動脈粥樣硬化及其他心血管疾病之慢性發炎性疾病的發病機制以及與諸如阿茲海默氏症(Alzheimer's disease)及帕金森氏病(Parkinson's disease)之退化性疾病有關。此外，多條證據支持氧化應力為正常衰老之主要機制的觀點。因此，對開發抑制氧化應力之治療存在需要。Oxidative stress and pathogenesis of chronic inflammatory diseases such as diabetes, cancer, atherosclerosis and other cardiovascular diseases, and degenerative effects such as Alzheimer's disease and Parkinson's disease Related to the disease. In addition, multiple lines of evidence support the view that oxidative stress is the primary mechanism of normal aging. Therefore, there is a need to develop a treatment for inhibiting oxidative stress.

使氧化應力降至最低被視為健康及長壽之關鍵。流行病學資料顯示富含抗氧化劑營養物之膳食賦予重大益處。除了典型抗氧化維生素(維生素C及E)外，膳食植物還含有許多似乎在食用後短期與長期降低氧化應力之多酚。已發展若干種試管內檢定來量測不同食品之抗氧化能力且多酚含量與抗氧化能力之間存在強相關性。Minimizing oxidative stress is considered a key to health and longevity. Epidemiological data show that diets rich in antioxidant nutrients confer significant benefits. In addition to the typical antioxidant vitamins (vitamins C and E), dietary plants also contain many polyphenols that appear to reduce oxidative stress in the short and long term after consumption. Several in-vitro assays have been developed to measure the antioxidant capacity of different foods and there is a strong correlation between polyphenol content and antioxidant capacity.

儘管包括葡萄之植物提供有前途之抗氧化劑來源，但人類群體廣泛使用植物化學物質存在顯著障礙。舉例而言，食品中活性劑之生體可用率對於用作抗氧化劑而言通常為次最佳的。活性劑亦可集中於植物之在供食用食物之典型製備中被忽視或丟棄之某些部分中。不同種類之相關食品亦可具有生物化學差異，此使得其對於治療用途並非最佳。Although plants including grapes provide a promising source of antioxidants, there are significant barriers to the widespread use of phytochemicals in the human population. For example, the bioavailability of an active agent in a food product is generally sub-optimal for use as an antioxidant. The active agent may also be concentrated in certain parts of the plant that are neglected or discarded in the typical preparation of the food for consumption. Different types of related foods may also have biochemical differences that make them less than optimal for therapeutic use.

本發明者已測定麝香葡萄果漿萃取物及日本蓼(Japanese knotweed)萃取物單獨或組合(呈混合物或膳食補充劑形式)之抗氧化能力，如藉由氧自由基吸收能力(Oxygen Radical Absorbance Capacity，ORAC)檢定所量測。同時量測樣品之親水性抗氧化能力與親脂性抗氧化能力。此等研究表明呈混合物或膳食補充劑形式之麝香葡萄果漿萃取物及日本蓼根萃取物在產生親脂性抗氧化能力方面具有強協同效應。未預料到在親脂性條件下展現選擇性協同作用。此等發現結果表明與任一單獨萃取物相比，含有日本蓼萃取物(至少作為實質上純白藜蘆醇之來源)與麝香葡萄果漿萃取物之混合物及膳食補充劑具有改良的抗氧化能力。The present inventors have determined the antioxidant capacity of muscadine pulp extract and Japanese knotweed extract alone or in combination (in the form of a mixture or dietary supplement), such as by oxygen radical absorption capacity (Oxygen Radical Absorbance Capacity). , ORAC) verification measured. At the same time, the hydrophilic antioxidant capacity and lipophilic antioxidant capacity of the sample were measured. These studies have shown that muscadine pulp extracts and Japanese root extracts in the form of a mixture or dietary supplement have a strong synergistic effect in producing lipophilic antioxidant capacity. It was not expected to exhibit selective synergy under lipophilic conditions. These findings indicate that the Japanese cockroach extract (at least as a source of substantially pure resveratrol) and the muscadine pulp extract mixture and dietary supplement have improved antioxidant capacity compared to either individual extract. .

本文揭示具有改良的抗氧化劑活性的組成物。在一些所揭示之具體實例中，該組成物包括具有至少2%之多酚含量的麝香葡萄(圓葉葡萄(Vitis rotundifolia ))果漿萃取物及來自非麝香葡萄來源(諸如日本蓼根萃取物)之最小純度為至少5%的白藜蘆醇，其中麝香葡萄多酚與反式白藜蘆醇之比率在0.1/1至10/1(重量/重量)之範圍內，由此提供具有抗氧化劑活性之組成物。在一特定實施例中，組成物包括具有至少2%之多酚含量的麝香葡萄(圓葉葡萄)果漿萃取物及來自非麝香葡萄來源(諸如日本蓼根萃取物)之最小純度為至少5%的白藜蘆醇，其中麝香葡萄果漿萃取物與白藜蘆醇之比率在0.2/1至50/1(重量/重量)，諸如5/1至50/1(重量/重量)或20/1至50/1(重量/重量)之範圍內，由此提供具有抗氧化劑活性之組成物。在一些實施例中，組成物具有每毫克多酚至少24微莫耳Trolox當量(微莫耳TE/毫克多酚)之總ORAC。Compositions having improved antioxidant activity are disclosed herein. In some disclosed embodiments, the composition comprises a muscadine ( Vitis rotundifolia ) pulp extract having a polyphenol content of at least 2% and a source derived from a non-musk grape (such as Japanese radix root extract) a minimum purity of at least 5% of resveratrol, wherein the ratio of muscadine polyphenol to trans-resveratrol is in the range of 0.1/1 to 10/1 (weight/weight), thereby providing resistance A composition of oxidant activity. In a particular embodiment, the composition comprises a muscadine (musk) pulp extract having a polyphenol content of at least 2% and a minimum purity of at least 5 from a non-musk source (such as Japanese jaundice extract). % resveratrol, wherein the ratio of muscadine pulp extract to resveratrol is from 0.2/1 to 50/1 (weight/weight), such as from 5/1 to 50/1 (weight/weight) or 20 Within the range of /1 to 50/1 (weight/weight), a composition having antioxidant activity is thus provided. In some embodiments, the composition has a total ORAC of at least 24 micromoles of Trolox equivalent per microgram of polyphenol (micromolar TE per milligram of polyphenol).

在一些實施例中，麝香葡萄(圓葉葡萄)果漿萃取物具有約4%之總多酚含量且組成物具有約18/1(重量/重量)之麝香葡萄果漿萃取物與反式白藜蘆醇的比率。在一實施例中，組成物具有約0.75/1(重量/重量)之麝香葡萄多酚與白藜蘆醇的比率。在某些實施例中，麝香葡萄果漿萃取物可包括於液體中之20%至50%固體。舉例而言，麝香葡萄果漿萃取物包含於液體中之約40%固體。在一些實施例中，白藜蘆醇包括自日本蓼(虎杖(Polygonum cuspidatum ))根萃取之至少98%反式白藜蘆醇。In some embodiments, the muscadine (musk) pulp extract has a total polyphenol content of about 4% and the composition has about 18/1 (weight/weight) of muscadine pulp extract and trans white The ratio of resveratrol. In one embodiment, the composition has a ratio of muscadine polyphenol to resveratrol of about 0.75/1 (weight/weight). In certain embodiments, the muscadine pulp extract may comprise from 20% to 50% solids in the liquid. For example, the muscadine pulp extract contains about 40% solids in the liquid. In some embodiments, the resveratrol comprises at least 98% trans-resveratrol extracted from the roots of Japanese cockroach ( Polygonum cuspidatum ).

在一些實施例中，所揭示之組成物可進一步包括接骨木萃取物、紫胡蘿蔔萃取物、賦形劑(諸如甘油、山梨糖醇、膠態二氧化矽或天然調味添加劑)或其組合。在一特定實施例中，組成物包括佔總組成物之23%至32%的麝香葡萄果漿萃取物、佔總組成物之1.2%至3.0%的白藜蘆醇、佔總組成物之0.12%至0.20%的紫胡蘿蔔萃取物、佔總組成物之0.4%至0.6%的接骨木萃取物、佔總組成物之50%至61%的山梨糖醇、佔總組成物之8%至12%的99.7%甘油組成物及佔總組成物之1%至2%的膠態二氧化矽。In some embodiments, the disclosed compositions can further include elderberry extract, purple carrot extract, excipients (such as glycerin, sorbitol, colloidal ceria or natural flavoring additives), or combinations thereof. In a particular embodiment, the composition comprises from 23% to 32% of the total composition of the muscadine pulp extract, 1.2% to 3.0% of the total composition of resveratrol, 0.12 of the total composition. % to 0.20% of purple carrot extract, 0.4% to 0.6% of total composition of elderberry extract, 50% to 61% of total composition of sorbitol, 8% to 12% of total composition % 99.7% glycerol composition and 1% to 2% of the total composition of colloidal cerium oxide.

在一更特定實施例中，組成物包括佔總組成物之約28.9%的麝香葡萄果漿萃取物(含有約4.1%多酚)、佔總組成物之約1.65%的白藜蘆醇、佔總組成物之約0.16%的紫胡蘿蔔萃取物、佔總組成物之約0.48%的接骨木萃取物、佔總組成物之約54.8%的山梨糖醇、佔總組成物之約10%的99.7%甘油組成物、佔總組成物之約2%的膠態二氧化矽、佔約1.27%之康考特葡萄(Concord grape)萃取物、佔約0.16%之解百納葡萄(Cabernet grape)萃取物及佔約0.48%之紅葡萄粉。In a more specific embodiment, the composition comprises about 28.9% of the total composition of the muscadine pulp extract (containing about 4.1% polyphenol), about 1.65% of the total composition of resveratrol, About 0.16% of the total composition of the purple carrot extract, about 0.48% of the total composition of the elderberry extract, about 54.8% of the total composition of sorbitol, about 10% of the total composition of 99.7 % glycerin composition, about 2% of the total composition of colloidal cerium oxide, about 1.27% of Concord grape extract, about 0.16% of Cabernet grape extract And about 0.48% of red grape powder.

在另一特定實施例中，組成物包括佔總組成物之約23.77%的麝香葡萄果漿萃取物(含有約5%多酚)、佔總組成物之約1.65%的白藜蘆醇、佔總組成物之約0.16%的紫胡蘿蔔萃取物、佔總組成物之約0.48%的接骨木萃取物、佔總組成物之約60%的山梨糖醇、佔總組成物之約10%的99.7%甘油組成物、佔總組成物之約2%的膠態二氧化矽、佔約1.27%之康考特葡萄萃取物、佔約0.16%之解百納葡萄萃取物及佔約0.48%之紅葡萄粉。In another specific embodiment, the composition comprises about 23.77% of the total composition of the muscadine pulp extract (containing about 5% polyphenol), about 1.65% of the total composition of resveratrol, About 0.16% of the total composition of the purple carrot extract, about 0.48% of the total composition of the elderberry extract, about 60% of the total composition of sorbitol, about 10% of the total composition of 99.7 % glycerin composition, about 2% of the total composition of colloidal cerium oxide, about 1.27% of Concord grape extract, about 0.16% of Cabernet grape extract and about 0.48% red Grape powder.

任一所揭示之組成物均可提供於呈液體或固體形式之非飲料食品、飲料或膳食補充劑中。此外，任一所揭示之組成物均可用作抗氧化劑，諸如抗衰老補充劑，包括皮膚改善劑。舉例而言，揭示防止或抑制細胞衰老之方法，其包括向有需要之個體投予足以防止或抑制一或多個與細胞衰老相關之過程之劑量的所揭示之組成物，該劑量諸如防止或抑制攝取該組成物之該個體體內的自由基形成或活性。Any of the disclosed compositions can be provided in a non-beverage food, beverage or dietary supplement in liquid or solid form. In addition, any of the disclosed compositions can be used as an antioxidant, such as an anti-aging supplement, including a skin improving agent. For example, a method of preventing or inhibiting cellular senescence is disclosed comprising administering to a subject in need thereof a disclosed composition sufficient to prevent or inhibit one or more dose-related processes associated with cellular senescence, such as preventing or Free radical formation or activity in the individual ingesting the composition is inhibited.

本文亦提供製備所揭示之抗氧化劑組成物的方法。此等方法可包括將具有至少2%之多酚含量的麝香葡萄(圓葉葡萄)果漿萃取物與來自非麝香葡萄來源之最小純度為至少5%反式白藜蘆醇的白藜蘆醇組合，其中麝香葡萄多酚與反式白藜蘆醇之比率在0.1/1至10/1(重量/重量)之範圍內，由此產生具有抗氧化劑活性之麝香葡萄果漿萃取物與反式白藜蘆醇的混合物。在一些實施例中，該等方法可包括將具有至少2%之多酚含量的麝香葡萄(圓葉葡萄)果漿萃取物與來自非麝香葡萄來源(諸如日本蓼根萃取物)之最小純度為至少5%反式白藜蘆醇的白藜蘆醇組合，其中麝香葡萄果漿萃取物與白藜蘆醇之比率在0.2/1至50/1(重量/重量)，諸如5/1至50/1(重量/重量)或20/1至50/1(重量/重量)之範圍內，包括18/1(重量/重量)，由此提供具有抗氧化劑活性之組成物。Also provided herein are methods of preparing the disclosed antioxidant compositions. Such methods may include muscadine (musk) pulp extract having a polyphenol content of at least 2% and resveratrol having a minimum purity of at least 5% trans-resveratrol from a non-musk source. a combination wherein the ratio of muscadine polyphenol to trans-resveratrol is in the range of 0.1/1 to 10/1 (weight/weight), thereby producing an antioxidant active muscadine pulp extract and trans A mixture of resveratrol. In some embodiments, the methods can include the minimum purity of the muscadine (lof grape) pulp extract having a polyphenol content of at least 2% and the non-musk source (such as Japanese jaundice extract). a resveratrol combination of at least 5% trans-resveratrol, wherein the ratio of muscadine pulp extract to resveratrol is from 0.2/1 to 50/1 (weight/weight), such as from 5/1 to 50 Within the range of /1 (weight/weight) or 20/1 to 50/1 (weight/weight), including 18/1 (weight/weight), thereby providing a composition having antioxidant activity.

所揭示之方法可進一步包括製備該麝香葡萄果漿萃取物，隨後將麝香葡萄果漿萃取物與反式白藜蘆醇組合。在一特定實施例中，製備麝香葡萄果漿萃取物包括將青銅色麝香葡萄果漿萃取物與紫色麝香葡萄果漿萃取物組合。在其他實施例中，製備麝香葡萄果漿萃取物包括自青銅色麝香葡萄果漿與紫色麝香葡萄果漿之混合物製備萃取物。The disclosed method can further comprise preparing the muscadine pulp extract, followed by combining the muscadine pulp extract with trans-resveratrol. In a particular embodiment, preparing the muscadine pulp extract comprises combining the bronze muscadine pulp extract with the purple muscadine pulp extract. In other embodiments, preparing the muscadine pulp extract comprises preparing a extract from a mixture of bronze muscadine pulp and purple muscadine pulp.

在一特定實施例中，該方法包括藉由將青銅色麝香葡萄果漿萃取物與紫色麝香葡萄果漿萃取物組合來製備麝香葡萄果漿萃取物，其中青銅色麝香葡萄果漿萃取物與紫色麝香葡萄果漿萃取物之比率在0.1至10(重量/重量)，諸如0.3至3(重量/重量)之範圍內。在一更特定實施例中，青銅色麝香葡萄果漿萃取物與紫色麝香葡萄果漿萃取物之比率為約2.25/1(重量/重量)。In a particular embodiment, the method comprises preparing a muscadine pulp extract by combining a bronze muscadine pulp extract with a purple muscadine pulp extract, wherein the bronze muscadine pulp extract is purple The ratio of the muscadine pulp extract is in the range of 0.1 to 10 (weight/weight), such as 0.3 to 3 (weight/weight). In a more specific embodiment, the ratio of bronze muscadine pulp extract to purple muscadine pulp extract is about 2.25/1 (weight/weight).

本揭示案之上述及其他特徵將自以下實施方式變得更顯而易見。The above and other features of the present disclosure will become more apparent from the following embodiments.

I.縮寫及術語：I. Abbreviations and terms:

(a)縮寫(a) abbreviation

FRAP： 漿之鐵還原能力 FRAP: Iron reduction ability of pulp

mg： 克 Mg: gram

ml： 升 Ml: liter

ORAC： 自由基吸收能力 ORAC: free radical absorption capacity

ROS： 性氧物質 ROS: Oxygen substances

TE： rolox當量 TE: rolox equivalent

TEAC： rolox當量抗氧化能力 TEAC: rolox equivalent antioxidant capacity

wt： 量 Wt: amount

(b)術語(b) terminology

提供以下對術語及方法之解釋以更好地描述本揭示案且在本揭示案之實施中指導一般技術者。如本文中及隨附申請專利範圍中所使用，除非本文另外明確規定，否則單數形式「一(a/an)」或「該(the)」包括複數個指示物。除非本文另外明確指示，否則術語「或(or)」係指所述二選一要素中之單個要素或兩個或兩個以上要素之組合。如本文中所使用，「包含(comprises)」意謂「包括(includes)」。因此，「包含A或B(comprising A or B)」意謂「包括A、B或A及B(including A,B,or A and B)」，而不排除其他要素。The following explanation of terms and methods are provided to better describe the present disclosure and to guide the general practitioner in the practice of the present disclosure. As used herein and in the appended claims, the singular "a" or "the" The term "or" means a single element or a combination of two or more elements of the two alternative elements, unless the context clearly indicates otherwise. As used herein, "comprises" means "includes". Therefore, "comprising A or B" means "including A, B or A and B" without excluding other elements.

除非另外說明，否則本文中所使用之所有技術及科學術語均具有與一般熟習本揭示案所屬技術者通常所理解相同之含義。儘管在本揭示案之實施或測試中可使用與本文所述類似或等效的方法及材料，但合適之方法及材料在下文中描述。另外應瞭解任何數值均為近似值，無論是否陳述「約(about/approximately)」一字或其類似字。除非另有指示，否則所有百分數及比率均以重量計算。All technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art of the present disclosure. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present disclosure, suitable methods and materials are described below. It should also be understood that any numerical value is an approximation, whether or not the word "about/approximately" or the like is stated. All percentages and ratios are by weight unless otherwise indicated.

投藥(administration)： 藉由任何有效途徑向個體提供或給與諸如包括麝香葡萄果漿萃取物及反式白藜蘆醇之組成物的藥劑。例示性投藥途徑包括(但不限於)口服、注射(諸如皮下、肌肉內、皮內、腹膜內、靜脈內及腫瘤內)、舌下、經皮、鼻內、局部及吸入途徑。 Administration: An agent such as a composition comprising muscadine pulp extract and trans-resveratrol is provided or administered to an individual by any effective means. Exemplary routes of administration include, but are not limited to, oral, injection (such as subcutaneous, intramuscular, intradermal, intraperitoneal, intravenous, and intratumoral), sublingual, transdermal, intranasal, topical, and inhalation routes.

抗氧化劑活性(antioxidant activity)： 包括清除及中和氧化自由基之活性。抗氧化劑活性可使用本文所揭示之方法以及此項技術中已知之方法來量測，該等方法包括氧自由基吸收能力(ORAC)檢定、血漿之鐵還原能力(Ferric Reducing Ability of Plasma，FRAP)檢定及Trolox當量抗氧化能力(Trolox Equivalent Antioxidant Capacity，TEAC)檢定。在一實施例中，若組成物具有每毫克多酚至少24微莫耳Trolox當量(微莫耳TE/毫克多酚)之總ORAC，則其具有抗氧化劑活性且可用作抗氧化劑。 Antioxidant activity: includes the activity of scavenging and neutralizing oxidative free radicals. Antioxidant activity can be measured using the methods disclosed herein and methods known in the art, including oxygen free radical absorption capacity (ORAC) assay, and ferric reducing ability of plasma (FRAP). Verification and Trolox Equivalent Antioxidant Capacity (TEAC) assay. In one embodiment, if the composition has a total ORAC of at least 24 micromoles of Trolox equivalent per microgram of polyphenol (micromolar TE per milligram of polyphenol), it has antioxidant activity and can be used as an antioxidant.

接骨木(elderberry)(西洋接骨木( Sambucus nigra ))： 一種屬於在歐洲及北美洲發現之五福花科(Adoxaceae)的植物，其具有若干區域性品種或亞種。花為平頭傘房花序。漿果為黑色至藍綠色且含有花青素(anthocyanin)及其他多酚系化合物，其中花青素及其他多酚系化合物之量及類型視品種而變化。 Elderberry ( Sambucus nigra ): A plant belonging to the family Adoxaceae found in Europe and North America, having several regional varieties or subspecies. The flower is a flat-headed umbrella inflorescence. Berries are black to blue-green and contain anthocyanins and other polyphenolic compounds, of which the amount and type of anthocyanins and other polyphenolic compounds vary depending on the species.

「接骨木萃取物(elderberry extract)」 可為藉由根據熟習此項技術者已知之任何萃取方法萃取接骨木所獲得之物質，只要其具有所需活性(例如，顏色穩定活性、抗氧化劑活性或其組合)即可。舉例而言，接骨木萃取物可包括藉由壓榨接骨木果實所獲得之果汁，或藉由根據熟習此項技術者習知之可選萃取方法及其類似方法萃取接骨木之完整果實或果實之皮或種子的合適部分所獲得之萃取物。同樣，接骨木果實之壓碎產物或經乾燥之接骨木果實濃縮物可作為「接骨木萃取物」使用。 "Elderberry extract" may be a substance obtained by extracting elderberry according to any extraction method known to those skilled in the art, as long as it has a desired activity (for example, color stabilizing activity, antioxidant activity, or The combination can be. For example, the elderberry extract may include juice obtained by pressing the elderberry fruit, or extracting the whole fruit or fruit of the elderberry by an optional extraction method and the like which are well known to those skilled in the art. Or an extract obtained from a suitable part of the seed. Similarly, crushed products of elderberry fruit or dried elderberry fruit concentrate can be used as "Sambucus extract".

賦形劑(excipient) ：一種用作組成物之活性成份之載劑的非活性物質。賦形劑可包括用於使具有極有效活性成份之調配物體積增大、允許習知及精確劑量、使該等活性成份穩定且使傳遞系統成為視覺及/或器官感覺上可接受的物質。醫藥賦形劑之實例包括澱粉、葡萄糖、乳糖、蔗糖、明膠、麥芽、稻米、麵粉、白堊、矽膠、硬脂酸鈉、單硬脂酸甘油酯、滑石、氯化鈉、脫脂奶粉、甘油、丙烯、乙二醇、水、乙醇及其類似物。在一特定實施例中，所揭示之抗衰老補充劑包括以下賦形劑：甘油、山梨糖醇、膠態二氧化矽及天然調味添加劑。 Excipient : An inactive substance used as a carrier for the active ingredient of a composition. Excipients can include materials for increasing the volume of a formulation having a very effective active ingredient, allowing for conventional and precise dosages, stabilizing the active ingredients, and rendering the delivery system visually and/or organoleptically acceptable. Examples of pharmaceutical excipients include starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, white peony, silicone, sodium stearate, glyceryl monostearate, talc, sodium chloride, skimmed milk powder, glycerin , propylene, ethylene glycol, water, ethanol and the like. In a particular embodiment, the disclosed anti-aging supplements include the following excipients: glycerin, sorbitol, colloidal ceria, and natural flavoring additives.

抑制(包括防止)細胞衰老(inhibiting(including preventing)cellular aging) ：抑制(例如防止)一或多個與細胞衰老相關之過程，諸如抑制攝取組成物之個體體內的自由基形成或活性。防止細胞衰老係指改善細胞衰老之徵象或症狀的干預。防止包括預防以延遲一或多個與細胞衰老相關之過程的發作。細胞衰老之阻止或抑制並不要求完全不存在細胞衰老。在一特定實施例中，所揭示之組成物使與細胞衰老相關之過程減少或延遲至少10%、至少20%、至少50%或甚至至少90%。該等減少可使用本文所揭示之方法以及此項技術中已知之方法來量測。 Inhibiting (including preventing) cellular aging : inhibiting (eg, preventing) one or more processes associated with cellular senescence, such as inhibiting free radical formation or activity in an individual ingesting a composition. Prevention of cellular aging refers to an intervention that improves the signs or symptoms of cellular aging. Prevention includes prevention to delay the onset of one or more processes associated with cellular aging. Blocking or inhibiting cellular senescence does not require complete absence of cellular senescence. In a particular embodiment, the disclosed compositions reduce or delay the process associated with cellular aging by at least 10%, at least 20%, at least 50%, or even at least 90%. Such reductions can be measured using the methods disclosed herein as well as methods known in the art.

麝香葡萄(Muscadine Grape)(圓葉葡萄) ：原產於美國東南部(southeastern United States)，且在自特拉華州(Delaware)至墨西哥灣(Gulf of Mexico)及密蘇里州(Missouri)、堪薩斯州(Kansas)、俄克拉荷馬州(Oklahoma)及德克薩斯州(Texas)以西之野外發現的葡萄。麝香葡萄很好地適應於美國東南部之溫熱、潮濕條件。果實長成3-40顆葡萄之小鬆散果穗，與歐洲及美洲葡萄之大緊密串特徵完全不同。圓形、1至1-1/2吋果實具有厚的硬皮且含有至多5粒硬的橢圓形籽。在顏色方面，果實在青銅綠色至青銅色、品紅色、紫色及近黑色範圍內。 Muscadine Grape (Clover) : native to southeastern United States, and from Delaware to the Gulf of Mexico and Missouri, Kansas Grapes found in the wilderness of the state (Kansas), Oklahoma, and Texas. Muscat is well adapted to the warm, humid conditions of the southeastern United States. The fruit grows into small loose ears of 3-40 grapes, which are completely different from the close string characteristics of European and American grapes. Round, 1 to 1-1/2 吋 fruit has a thick crust and contains at most 5 hard oval seeds. In terms of color, the fruit is in the range of bronze green to bronze, magenta, purple and near black.

可得到許多不同品種之麝香葡萄，包括雌性(只有雌蕊)品種，諸如黑美人(Black Beauty)、黑仔(Black Fry)、達琳(Darlene)、小魚(Fry)、希金斯(Higgins)、珍寶(Jumbo)、斯卡珀農(Scuppernong)、糖門(Sugargate)、高峰(Summit)、皇冠(Supreme)及甜珍妮(Sweet Jenny)；及自花受精品種，諸如卡洛斯(Carlos)、考瓦特(Cowart)、迪克西蘭(Dixieland)、繁星紅(Dixie Red)、小魚無籽(Fry Seedless)、木蘭(Magnolia)、內斯比特(Nesbitt)、貴族(Noble)、紅門(Redgate)、瑞格爾(Regale)及思德琳(Sterling)。Many different varieties of muscat grape are available, including female (only pistil) varieties, such as Black Beauty, Black Fry, Darlene, Fry, Higgins. Jumbo, Scuppernong, Sugargate, Summit, Supreme and Sweet Jenny; and self-cultivating species such as Carlos, Cowart, Dixieland, Dixie Red, Fry Seedless, Magnolia, Nesbitt, Noble, Redgate ), Regale and Sterling.

舉例而言，麝香葡萄之青銅色(bronze colored )品種由熟習此項技術者鑑別為包括卡洛斯、曹旺(Chowan)、多琳(Doreen)、希金斯、木蘭、勿忘(Nevermiss)、帕姆利科(Pamlico)、羅諾克(Roanoke)、斯卡珀農、思德琳及高峰栽培品種。紫色至黑色(purple to black colored) 品種包括阿爾伯馬爾(Albermarle)、豐盛(Bountiful)、考瓦特、GA-1、亨特(Hunt)、NC-1、貴族、瑞格爾、北卡羅萊那(Tarheel)及珍寶。For example, the bronze colored varieties of muscat are identified by those skilled in the art as including Carlos, Chowan, Doreen, Higgins, Magnolia, Nevermiss, Pamlico, Roanoke, Scarperon, Sterling and peak cultivars. Purple to black colored varieties include Albermarle, Bountiful, Cowart, GA-1, Hunt, NC-1, Noble, Regal, North Carolina (Tarheel) and treasures.

完整麝香葡萄之植物化學物組份不同於歐洲葡萄(Vitis vinifera )。麝香葡萄具有特徵為高土耳其鞣酸糖苷、五倍子酸糖苷及類黃酮苷濃度之較高總酚系化合物含量。麝香葡萄中存在土耳其鞣酸(ellagic acid)很獨特且土耳其鞣酸以游離土耳其鞣酸、土耳其鞣酸糖苷、甲氧基化衍生物及並沒食子鞣質(ellagitannin)之形式存在。另一獨特特徵為麝香葡萄中觀察到花青素化學性質。以呈非醯化形式之飛燕草素(delphinidin)、花色素(cyanidin)、牽牛花素(petunidin)、芍藥色素(peonidin)及錦葵色素(malvidin)之3,5-二糖苷(與3-糖苷相對)形式存在的此等化合物及受葡萄中所存在之其他花青素的天然顏色影響使汁液及果漿呈深紫色。紫色果漿萃取物含有花青素，而青銅色果漿萃取物並不含花青素。The phytochemical component of the complete muscat grape is different from the Vitis vinifera . Musk grape has a higher total phenolic compound content characterized by high concentrations of citric acid glycosides, gallic acid glycosides and flavonoid glycosides. The presence of turkey ellagic acid in muscadine is unique and Turkish citric acid is present in the form of free Turkish citric acid, Turkish citrate glycosides, methoxylated derivatives and ellagitannin. Another unique feature is the anthocyanin chemistry observed in muscadine. 3,5-diglucoside (with 3-in a non-deuterated form of delphinidin, cyanidin, petunidin, peonidin, and malvidin) The presence of such compounds in the form of glycosides and the natural color of other anthocyanins present in the grapes render the juice and pulp dark purple. The purple pulp extract contains anthocyanins, while the bronze pulp extract does not contain anthocyanins.

紅色及紫色花青素為具有抗氧化劑性質之多酚化合物。黃酮醇為完整麝香葡萄中所存在之第二最豐富的類黃酮。關於麝香葡萄皮部分所報導之主要酚系化合物(按遞減順序)為土耳其鞣酸、楊梅黃酮(myricetin)、槲皮素(quercetin)及堪非黃酮醇(kaempferol)，而關於於籽所報導之主要酚系化合物為表兒茶素(epicatechin)、兒茶素(catechin)及五倍子酸(gallic acid)(Patrana-Bonilla等人J. Agric. Food Chem . 51:5497-5503,2003)。Red and purple anthocyanins are polyphenolic compounds having antioxidant properties. Flavonol is the second most abundant flavonoid in intact muscat grape. The main phenolic compounds reported in the muscadine section (in descending order) are turmeric, myricetin, quercetin and kaempferol, as reported by the seed. The main phenolic compounds are epicatechin, catechin and gallic acid (Patrana-Bonilla et al . J. Agric. Food Chem . 51: 5497-5503, 2003).

醫藥學上可接受之媒劑(Pharmaceutically Acceptable Vehicles) ：適用於本揭示案之醫藥學上可接受之媒劑(載劑)為習知的。Remington’s Pharmaceutical Sciences ,E. W. Martin,Mack Publishing公司,Easton,PA,第19版(1995)描述適合於醫藥傳遞一或多種組成物(諸如一或多種麝香葡萄組成物)及其他醫藥劑之組成物及調配物。 Pharmaceutically Acceptable Vehicles : Pharmaceutically acceptable vehicles (carriers) suitable for use in the present disclosure are conventional. Remington's Pharmaceutical Sciences , EW Martin, Mack Publishing Company, Easton, PA, 19th edition (1995) describes compositions and formulations suitable for the delivery of one or more compositions (such as one or more muscadine compositions) and other pharmaceutical agents for pharmaceutical delivery. Things.

一般而言，該媒劑之性質將視所使用之特定投藥模式而定。舉例而言，非經腸調配物通常包含作為媒劑之可注射流體，包括醫藥學上及生理學上可接受之流體，諸如水、生理食鹽水、平衡鹽溶液、右旋糖水溶液、甘油或其類似物。對於固體組成物(例如，散劑、丸劑、錠劑或膠囊形式)而言，習知無毒固體媒劑可包括(例如)醫藥級甘露糖醇、乳糖、澱粉或硬脂酸鎂。除生物中性媒劑外，欲投予之醫藥組成物亦可含有少量無毒輔助物質，諸如濕潤劑或乳化劑、防腐劑及pH值緩衝劑及其類似物，例如乙酸鈉或脫水山梨糖醇單月桂酸酯。In general, the nature of the vehicle will depend on the particular mode of administration employed. For example, parenteral formulations typically comprise an injectable fluid as a vehicle, including pharmaceutically and physiologically acceptable fluids such as water, physiological saline, balanced salt solutions, aqueous dextrose, glycerol or Its analogues. For solid compositions (eg, powders, pills, troches, or capsules), conventional non-toxic solid vehicles may include, for example, pharmaceutical grade mannitol, lactose, starch, or magnesium stearate. In addition to the biologically neutral vehicle, the pharmaceutical composition to be administered may also contain small amounts of non-toxic auxiliary substances such as wetting or emulsifying agents, preservatives and pH buffers and the like, such as sodium acetate or sorbitan. Monolaurate.

經純化(purified) ：術語經純化並不要求絕對純度；更確切而言，希望其為相對術語。因此，舉例而言，經純化物質為與該物質在其天然環境中(例如在果實(例如葡萄)中)相比物質更富集之物質。在一具體實例中，製劑經純化以使物質佔製劑總含量之至少約5%(諸如(但不限於)至少10%、20%、30%、40%、50%、70%、80%、90%、95%、98%或99%)。在一實施例中，所揭示之具有抗氧化劑活性的組成物包括最小純度為總白藜蘆醇製劑之至少50%、70%、80%、90%、95%、98%或99%(重量)的反式白藜蘆醇。 Purified : The term purified does not require absolute purity; rather, it is a relative term. Thus, for example, a purified material is one that is more enriched with the substance in its natural environment (eg, in a fruit (eg, grape)). In one embodiment, the formulation is purified such that the material comprises at least about 5% of the total formulation (such as, but not limited to, at least 10%, 20%, 30%, 40%, 50%, 70%, 80%, 90%, 95%, 98% or 99%). In one embodiment, the disclosed composition having antioxidant activity comprises a minimum purity of at least 50%, 70%, 80%, 90%, 95%, 98% or 99% of the total resveratrol formulation (weight Trans-resveratrol.

果漿(pomace)： 在汁液萃取後剩餘之皮、籽及果肉。在一實施例中，果漿萃取物為青銅色麝香葡萄果漿萃取物、紫色麝香葡萄果漿萃取物或其組合。許多不同品種之麝香葡萄果漿可用作起始物質，且其包括雌性(只有雌蕊的)品種，諸如黑美人、黑仔、達琳、小魚、希金斯、珍寶、斯卡珀農、糖門、高峰、皇冠及甜珍妮，及自花受精品種，諸如卡洛斯、考瓦特、迪克西蘭、繁星紅、小魚無籽、木蘭、內斯比特、貴族、紅門、瑞格爾及思德琳。 Pomace: The skin, seeds and pulp remaining after juice extraction. In one embodiment, the pulp extract is a bronze muscadine pulp extract, a purple muscadine pulp extract, or a combination thereof. Many different varieties of muscadine pulp can be used as a starting material, and include female (only pistil) varieties, such as black beauty, black babies, Darin, small fish, Higgins, treasure, Scarperon, Sugar Gate, Peak, Crown and Sweet Jenny, and self-cultivating species such as Carlos, Cowart, Dixieland, Starred Red, Small Fish Seedless, Magnolia, Nesbitt, Noble, Red Gate, Regal and Sidley.

麝香葡萄果漿含有具有抗氧化劑性質之酚系化合物，包括五倍子酸及土耳其鞣酸。Muscat grape pulp contains phenolic compounds with antioxidant properties, including gallic acid and turmeric tannin.

紫胡蘿蔔(purple carrot)(胡蘿蔔( Daucus carota ))： 一種含有花青素色素之胡蘿蔔的栽培品種。「紫胡蘿蔔萃取物(purple carrot extract)」可為藉由根據熟習此項技術者已知之任何萃取方法來萃取紫胡蘿蔔所獲得之物質，只要其具有所需活性(例如，顏色穩定活性、抗氧化劑活性或其組合)即可。在一實施例中，紫胡蘿蔔萃取物具有使麝香葡萄顏色色素穩定之性質且因此可隨後用作顏色穩定添加劑。 Purple carrot ( Daucus carota ): A cultivar of carrots containing anthocyanin pigments. "Purple carrot extract" may be a substance obtained by extracting purple carrots according to any extraction method known to those skilled in the art as long as it has a desired activity (for example, color stabilizing activity, antioxidants) The activity or a combination thereof can be used. In one embodiment, the purple carrot extract has the property of stabilizing the muscat color pigment and thus can be subsequently used as a color stabilizing additive.

白藜蘆醇(resveratrol)： 一種為二苯乙烯類化合物(stilbenoid)(二苯乙烯之衍生物)之植物防禦素(phytoalexin)，且藉助於酶二苯乙烯合成酶在植物體內產生。白藜蘆醇以兩種結構異構體形式存在：順式白藜蘆醇及反式白藜蘆醇。當加熱或曝露於紫外線照射時，反式白藜蘆醇可經歷異構化轉變為順式。 Resveratrol: A phytoalexin which is a stilbenoid (a derivative of stilbene) and is produced in plants by means of an enzyme stilbene synthase. Resveratrol exists in two structural isoforms: cis resveratrol and trans resveratrol. Trans-resveratrol can undergo isomerization to cis when heated or exposed to ultraviolet radiation.

白藜蘆醇以廣泛變化的量存在於葡萄、樹莓、桑椹、李子、花生、越橘屬(Vaccinium)物種之漿果(包括藍莓、歐洲越橘及大果蔓越橘)、一些松類(諸如歐洲赤松(Scots pine)及北美白松(eastern white pine))及虎杖及日本蓼之根及莖中。在葡萄中，白藜蘆醇主要存在於皮及籽中。存在於葡萄皮中之白藜蘆醇的量隨葡萄栽培品種、其地理來源及暴露於真菌感染而變化。Resveratrol is present in widely varying amounts in berries, raspberries, mulberries, plums, peanuts, berries of the Vaccinium species (including blueberries, bilberry and cranberries) and some pines ( Such as the European Scots pine and the eastern white pine, and the roots and stems of Polygonum cuspidatum and Japanese cockroach. In grapes, resveratrol is mainly found in the skin and seeds. The amount of resveratrol present in the grape skin varies with the grape cultivar, its geographic origin, and exposure to fungal infections.

如本文中所使用，術語白藜蘆醇可包括自諸如葡萄之植物萃取的天然反式白藜蘆醇或合成反式白藜蘆醇。如本文中所使用，術語白藜蘆醇可包括反式白藜蘆醇之改質調配物，諸如微囊封或水分散性形式。As used herein, the term resveratrol may include natural trans-resveratrol or synthetic trans-resveratrol extracted from a plant such as grapes. As used herein, the term resveratrol can include a modified formulation of trans-resveratrol, such as a microencapsulated or water-dispersible form.

個體(subject)： 活的多細胞脊椎生物體，包括人類與獸類個體之類別。 Subject: A living multi-cellular vertebrate organism, including the categories of human and veterinary individuals.

治療有效量(Therapeutically Effective Amount)： 組成物單獨或與另一(其他)藥劑(例如其他抗氧化劑)一起誘發所需反應(例如，防止或抑制細胞衰老)之量。本文所揭示之製劑可以治療有效量投予。 Therapeically Effective Amount: The amount of a composition that, alone or in combination with another (other) agent (eg, other antioxidants), induces a desired response (eg, prevents or inhibits cellular senescence). The formulations disclosed herein can be administered in a therapeutically effective amount.

在一實施例中，所需反應為抑制或降低與該組成物所投予之個體體內的一或多個細胞衰老過程相關之自由基產生或活性。關於組成物之有效性，自由基產生或活性並不需要完全被消除。舉例而言，與在組成物不存在情況下之自由基產生或活性相比，組成物可使產生或活性降低所需量，例如至少10%、至少20%、至少50%、至少60%、至少70%、至少80%、至少90%、至少95%或至少98%。In one embodiment, the desired response is to inhibit or reduce free radical production or activity associated with one or more cellular senescence processes in the subject to which the composition is administered. Regarding the effectiveness of the composition, free radical production or activity does not need to be completely eliminated. For example, the composition can reduce the production or activity by a desired amount, such as at least 10%, at least 20%, at least 50%, at least 60%, compared to free radical production or activity in the absence of the composition. At least 70%, at least 80%, at least 90%, at least 95% or at least 98%.

在治療過程中，治療有效量之所揭示麝香葡萄果漿萃取物組成物可以單一劑量或以若干劑量投予，例如每日投予。然而，該治療有效量可視所治療之個體、所治療病狀的嚴重程度及類型及投藥方式而定。舉例而言，若經口投予，則治療有效量之該等藥劑可在每日約1毫克至約500毫克反式白藜蘆醇、麝香葡萄多酚或其組合範圍內變化。During treatment, a therapeutically effective amount of the disclosed muscadine pulp extract composition can be administered in a single dose or in several doses, such as daily administration. However, the therapeutically effective amount will depend on the individual being treated, the severity and type of condition being treated, and the mode of administration. For example, if administered orally, a therapeutically effective amount of such agents can vary from about 1 mg to about 500 mg per day of trans-resveratrol, muscadine polyphenols, or combinations thereof.

單位劑量(unit dose)： 含有經計算個別或共同產生所需作用(諸如治療作用)之預定量之活性物質的物理離散單位。可使用單一單位劑量或複數個單位劑量來提供所需作用或活性，諸如抗氧化劑活性。在一實施例中，單位劑量包括防止或抑制一或多個與細胞衰老相關之過程的所需量之藥劑。 Unit dose: A physical discrete unit containing a predetermined amount of active substance calculated to produce a desired effect, such as a therapeutic effect, individually or collectively. A single unit dose or a plurality of unit doses can be used to provide the desired effect or activity, such as antioxidant activity. In one embodiment, the unit dose includes a desired amount of the agent that prevents or inhibits one or more processes associated with cellular aging.

II.若干具體實例之描述II. Description of several specific examples

已充分研究植物藥劑作為可用於降低氧化應力之營養藥劑的潛在來源。紅葡萄為已成為廣泛研究之對象的許多食品之一。紅葡萄之抗氧化劑作用已廣泛歸因於多酚系化合物，諸如白藜蘆醇及原花色素(procyanidin)，其中後者以實質量出現於葡萄中。Plant agents have been well studied as potential sources of nutrients that can be used to reduce oxidative stress. Red grapes are one of many foods that have been the subject of extensive research. The antioxidant action of red grapes has been widely attributed to polyphenolic compounds such as resveratrol and procyanidin, the latter of which appear in grapes in substantial quality.

本發明者進行一系列研究以開發一種足以提高土耳其鞣酸之最大溶解度的麝香葡萄果漿萃取物。現已發現未經濃縮之65%/35%之青銅色麝香葡萄果漿萃取物與紫色麝香葡萄果漿萃取物(體積/體積)的比率提高土耳其鞣酸在兩種果漿萃取物之間的最大溶解度。The inventors conducted a series of studies to develop a muscadine pulp extract sufficient to increase the maximum solubility of citric acid in Turkey. It has been found that the ratio of unconcentrated 65%/35% bronze muscadine pulp extract to purple muscadine pulp extract (vol/vol) increases the amount of turmeric tannin between the two pulp extracts. Maximum solubility.

土耳其鞣酸(其為麝香葡萄特有的)在麝香葡萄果漿萃取物中之最大溶解度的測定允許製備具有提高之土耳其鞣酸溶解度的萃取物(且因此捕獲果漿之多酚分布)，此又允許製備具有改良的抗氧化劑活性的萃取物。因此，本文揭示具有改良的土耳其鞣酸溶解度之麝香葡萄果漿萃取物及使用該等萃取物之方法。舉例而言，所揭示之麝香葡萄萃取物可以非飲料食品、飲料、膳食補充劑或局部軟膏形式單獨投予或與其他化合物組合投予。亦使製備麝香葡萄果漿萃取物之方法成為可能，該方法包括將青銅色麝香葡萄果漿萃取物與紫色麝香葡萄果漿萃取物組合以產生麝香葡萄果漿萃取物，其中青銅色麝香葡萄果漿萃取物與紫色麝香葡萄果漿萃取物之比率在0.1至10(重量/重量)，諸如0.3至3(重量/重量)之範圍內。The determination of the maximum solubility of citric acid in Turkey (which is unique to muscat) in muscadine pulp extracts allows the preparation of extracts with increased solubility of citric acid in Turkey (and thus the polyphenol distribution of the pulp), which in turn It is allowed to prepare an extract with improved antioxidant activity. Accordingly, the present invention discloses muscadine pulp extracts having improved Turkish citric acid solubility and methods of using such extracts. For example, the disclosed muscadine extract can be administered alone or in combination with other compounds in the form of a non-beverage food, beverage, dietary supplement or topical ointment. A method of preparing a muscadine pulp extract is also possible, the method comprising combining a bronze muscadine pulp extract with a purple muscadine pulp extract to produce a muscadine pulp extract, wherein the bronze muscat fruit The ratio of the pulp extract to the purple muscadine pulp extract is in the range of 0.1 to 10 (weight/weight), such as 0.3 to 3 (weight/weight).

另外，本發明者測定所揭示之麝香葡萄果漿萃取物及日本蓼萃取物(標準化為98%反式白藜蘆醇)單獨或組合(以混合物或膳食補充劑形式)的抗氧化能力，如藉由ORAC檢定所量測。同時量測樣品之親水性抗氧化能力與親脂性抗氧化能力。此等研究表明呈混合物或膳食補充劑形式之麝香葡萄果漿萃取物及日本蓼根萃取物(標準化為98%反式白藜蘆醇)在產生親脂性抗氧化能力方面具有強協同效應。未預料到在親脂性條件下展現選擇性協同作用。同樣，雖然組成物中所用之麝香葡萄萃取物可為天然萃取物，其會因物種、萃取方法等而變化，只要維持所揭示之麝香葡萄多酚與白藜蘆醇的比率即可，但協同親脂性抗氧化劑活性得到保留。In addition, the inventors determined the antioxidant capacity of the disclosed muscadine pulp extract and Japanese cockroach extract (standardized to 98% trans-resveratrol) alone or in combination (in the form of a mixture or a dietary supplement), such as Measured by ORAC test. At the same time, the hydrophilic antioxidant capacity and lipophilic antioxidant capacity of the sample were measured. These studies have shown that muscadine grape fruit extract in the form of a mixture or dietary supplement and Japanese root extract (standardized to 98% trans-resveratrol) have a strong synergistic effect in producing lipophilic antioxidant capacity. It was not expected to exhibit selective synergy under lipophilic conditions. Similarly, although the muscadine extract used in the composition may be a natural extract, it may vary depending on the species, the extraction method, etc., as long as the ratio of the disclosed muscadine polyphenol to resveratrol is maintained, but synergistically Lipophilic antioxidant activity is retained.

此等發現結果表明與任一單獨萃取物相比，含有日本蓼萃取物(標準化為98%反式白藜蘆醇)與麝香葡萄果漿萃取物兩者之混合物及膳食補充劑具有改良的親脂性抗氧化能力。重要的是，發生於親脂性環境中之氧化過程視為造成及/或引發多種疾病病況之發病機制，諸如動脈粥樣硬化中之低密度脂蛋白(low density lipoprotein，LDL)氧化及第II型糖尿病中之肥胖誘發性胰島素抗性。此外，膳食脂質在胃腸道中發生氧化導致諸如丙二醛(malondialdehyde，MDA)之細胞毒性及基因毒性脂質過氧化產物的吸收。因此，所揭示之具有高親脂性抗氧化能力的組成物可在抵消上述脂質氧化之有害後果方面賦予有益作用。These findings indicate that the mixture containing Japanese cockroach extract (standardized as 98% trans-resveratrol) and muscadine pulp extract and dietary supplements has improved progeny compared to either individual extract. Lipid antioxidant capacity. Importantly, the oxidative process occurring in a lipophilic environment is thought to cause and/or trigger the pathogenesis of a variety of disease conditions, such as low density lipoprotein (LDL) oxidation in atherosclerosis and type II Obesity-induced insulin resistance in diabetes. In addition, oxidation of dietary lipids in the gastrointestinal tract results in absorption of cytotoxicity such as malondialdehyde (MDA) and genotoxic lipid peroxidation products. Therefore, the disclosed composition having high lipophilic antioxidant ability can impart a beneficial effect in counteracting the harmful effects of the above lipid oxidation.

此發現使具有改良的抗氧化劑活性之麝香葡萄果漿萃取物組成物及製備該等組成物之方法成為可能。舉例而言，本文揭示具有抗氧化劑活性之麝香葡萄果漿萃取物組成物，其可以非飲料食品、飲料、液體或固體膳食補充劑或局部軟膏形式投予。製備所揭示之組成物的方法可包括將具有至少2%之多酚含量的麝香葡萄(圓葉葡萄)果漿萃取物與來自非麝香葡萄來源之最小純度為至少5%的反式白藜蘆醇組合，其中麝香葡萄多酚與反式白藜蘆醇之比率在0.1/1至10/1(重量/重量)之範圍內，由此產生具有抗氧化劑活性之麝香葡萄果漿萃取物與反式白藜蘆醇的混合物。製備所揭示之組成物的方法亦可包括將具有至少2%之多酚含量的麝香葡萄(圓葉葡萄)果漿萃取物與來自非麝香葡萄來源之最小純度為至少5%的反式白藜蘆醇組合，其中麝香葡萄果漿萃取物與反式白藜蘆醇之比率在0.2/1至50/1(重量/重量)，諸如5/1至50/1(重量/重量)，包括20/1至50/1(重量/重量)之範圍內，諸如18/1(重量/重量)，由此產生具有抗氧化劑活性之麝香葡萄果漿萃取物與反式白藜蘆醇的混合物。This finding made it possible to make muscadine pulp extract compositions with improved antioxidant activity and methods of preparing such compositions. For example, disclosed herein is a muscadine pulp extract composition having antioxidant activity that can be administered in the form of a non-beverage food, beverage, liquid or solid dietary supplement or topical ointment. The method of preparing the disclosed composition may comprise adding a muscadine (circle grape) pulp extract having a polyphenol content of at least 2% to a trans-white cane having a minimum purity of at least 5% from a non-musk source. An alcohol combination in which the ratio of muscadine polyphenol to trans-resveratrol is in the range of 0.1/1 to 10/1 (weight/weight), thereby producing an antioxidant activity of muscadine pulp extract and anti-oxidant a mixture of resveratrol. The method of preparing the disclosed compositions may also comprise extracting a muscadine (lof grape) pulp extract having a polyphenol content of at least 2% with a trans-white peony having a minimum purity of at least 5% from a non-musk source. a rosin combination wherein the ratio of the muscadine pulp extract to trans-resveratrol is from 0.2/1 to 50/1 (weight/weight), such as from 5/1 to 50/1 (weight/weight), including 20 Within the range of /1 to 50/1 (weight/weight), such as 18/1 (weight/weight), thereby producing a mixture of muscadine pulp extract and trans-resveratrol having antioxidant activity.

已發現親脂性抗氧化劑有效防止各種類型之皮膚損傷，諸如(但不限於)藉由抑制脂質過氧化及由脂質過氧化產生之產物(諸如交聯劑)。已知氧化應力為造成正常衰老之主要機制。本文中顯示所揭示之組成物具有改良的親脂性抗氧化劑活性。基於此等觀察結果，揭示抑制細胞衰老(諸如防止或抑制自由基產生或活性)之方法。該等方法包括使用所揭示之具有改良的抗氧化能力的組成物(例如膳食補充劑)作為抗衰老補充劑。舉例而言，使防止或減少攝取混合物或膳食補充劑組成物之個體體內的一或多個與細胞衰老相關之過程(諸如降低或抑制自由基產生或活性)的方法成為可能。Lipophilic antioxidants have been found to be effective in preventing various types of skin damage such as, but not limited to, products that inhibit lipid peroxidation and are produced by lipid peroxidation, such as crosslinkers. Oxidative stress is known to be the primary mechanism responsible for normal aging. The compositions disclosed herein are shown to have improved lipophilic antioxidant activity. Based on these observations, methods for inhibiting cellular senescence, such as preventing or inhibiting free radical production or activity, are disclosed. Such methods include the use of the disclosed compositions having improved antioxidant capacity (e.g., dietary supplements) as an anti-aging supplement. For example, a method of preventing or reducing one or more processes associated with cellular aging, such as reducing or inhibiting free radical production or activity, in an individual ingesting a mixture or dietary supplement composition is made possible.

A. 　麝香葡萄果漿萃取物 A. Muscat grape puree extract

本文揭示來源於青銅色麝香葡萄果漿及紫色麝香葡萄果漿之麝香葡萄果漿萃取物。在一些具體實例中，麝香葡萄果漿萃取物中青銅色麝香葡萄果漿萃取物與紫色麝香葡萄果漿萃取物之比率在0.1至10(重量/重量)，諸如0.3至3(重量/重量)之範圍內。舉例而言，青銅色麝香葡萄果漿萃取物與紫色麝香葡萄果漿萃取物之比率為約2.75/約1(重量/重量)、2.5/約1(重量/重量)、約2.25/約1(重量/重量)、約2/約1(重量/重量)、約1.5/約1(重量/重量)或約1/約1(重量/重量)。如本文中所使用，術語「約(about)」定義為±0.5。在一特定實施例中，青銅色麝香葡萄果漿萃取物與紫色麝香葡萄果漿萃取物之比率為約2.25/約1(重量/重量)。This article discloses extracts of muscadine pulp derived from bronze muscadine pulp and purple musk grape pulp. In some embodiments, the ratio of the bronze muscadine pulp extract to the purple muscadine pulp extract in the muscadine pulp extract is from 0.1 to 10 (weight/weight), such as from 0.3 to 3 (weight/weight). Within the scope. For example, the ratio of bronze muscadine pulp extract to purple muscadine pulp extract is about 2.75 / about 1 (weight / weight), 2.5 / about 1 (weight / weight), about 2.25 / about 1 ( Weight/weight), about 2/about 1 (weight/weight), about 1.5/about 1 (weight/weight) or about 1/about 1 (weight/weight). As used herein, the term "about" is defined as ±0.5. In a particular embodiment, the ratio of bronze muscadine pulp extract to purple muscadine pulp extract is about 2.25 / about 1 (weight / weight).

在某些具體實例中，所揭示之麝香葡萄(圓葉葡萄)果漿萃取物具有至少2%之多酚含量。舉例而言，多酚含量為至少3%、至少3.5%、至少4%、至少4.5%、至少5%、至少6%、至少8%、至少10%、至少12%或至少14%。在一特定實施例中，麝香葡萄(圓葉葡萄)果漿萃取物具有約4%之多酚含量。In certain embodiments, the disclosed muscadine (loft grape) pulp extract has a polyphenol content of at least 2%. For example, the polyphenol content is at least 3%, at least 3.5%, at least 4%, at least 4.5%, at least 5%, at least 6%, at least 8%, at least 10%, at least 12%, or at least 14%. In a particular embodiment, the muscadine (circle grape) pulp extract has a polyphenol content of about 4%.

在一些具體實例中，所揭示之麝香葡萄果漿萃取物包括於液體中之20%至50%固體，諸如至少25%、至少30%、至少35%、至少37%、至少40%、至少42%、至少44%、至少46%或至少48%。在一特定實施例中，萃取物包括於液體中之約40%固體。In some embodiments, the disclosed muscadine pulp extract comprises 20% to 50% solids in a liquid, such as at least 25%, at least 30%, at least 35%, at least 37%, at least 40%, at least 42 %, at least 44%, at least 46% or at least 48%. In a particular embodiment, the extract comprises about 40% solids in the liquid.

B.　製備麝香葡萄果漿萃取物之方法B. Method for preparing muscadine pulp extract

亦揭示製備麝香葡萄果漿萃取物之方法，其中青銅色麝香葡萄果漿萃取物與紫色麝香葡萄果漿萃取物之比率在0.1至10(重量/重量)，諸如0.3至3(重量/重量)之範圍內。在一所揭示之具體實例中，藉由將青銅色麝香葡萄果漿萃取物與紫色麝香葡萄果漿萃取物組合來製備麝香葡萄果漿萃取物。在一特定實施例中，藉由將青銅色麝香葡萄果漿萃取物與紫色麝香葡萄果漿萃取物組合來製備麝香葡萄果漿萃取物，其中青銅色麝香葡萄果漿萃取物與紫色麝香葡萄果漿萃取物之比率在0.1至10(重量/重量)，諸如0.3至3(重量/重量)之範圍內，包括約2.75/約1(重量/重量)、2.5/約1(重量/重量)、約2.25/約1(重量/重量)、約2/約1(重量/重量)、約1.5/約1(重量/重量)或約1/約1(重量/重量)。在一甚至更特定實施例中，藉由將青銅色麝香葡萄果漿萃取物與紫色麝香葡萄果漿萃取物組合來製備麝香葡萄果漿萃取物，其中青銅色麝香葡萄萃取物與紫色麝香葡萄果漿萃取物之比率為約2.25/1(重量/重量)。Also disclosed is a method of preparing a muscadine pulp extract, wherein the ratio of the bronze muscadine pulp extract to the purple muscadine pulp extract is from 0.1 to 10 (weight/weight), such as from 0.3 to 3 (weight/weight). Within the scope. In a disclosed embodiment, the muscadine pulp extract is prepared by combining a bronze muscadine pulp extract with a purple muscadine pulp extract. In a particular embodiment, the muscadine pulp extract is prepared by combining a bronze muscadine pulp extract with a purple muscadine pulp extract, wherein the bronze muscadine pulp extract and the purple muscat fruit The ratio of the pulp extract is in the range of 0.1 to 10 (weight/weight), such as 0.3 to 3 (weight/weight), including about 2.75/about 1 (weight/weight), 2.5/about 1 (weight/weight), About 2.25 / about 1 (weight / weight), about 2 / about 1 (weight / weight), about 1.5 / about 1 (weight / weight) or about 1 / about 1 (weight / weight). In an even more specific embodiment, the muscadine pulp extract is prepared by combining a bronze muscadine pulp extract with a purple muscadine pulp extract, wherein the bronze muscat grape extract and the purple muscat fruit The ratio of the pulp extract was about 2.25/1 (weight/weight).

在一些實施例中，製備麝香葡萄果漿萃取物之方法可包括分別製備青銅色麝香葡萄果漿萃取物及紫色麝香葡萄果漿萃取物。舉例而言，分別用水(較佳為熱水)萃取青銅色麝香葡萄果漿及紫色麝香葡萄果漿。製備青銅色麝香葡萄果漿萃取物及紫色麝香葡萄果漿萃取物可進一步包括使青銅色麝香葡萄果漿萃取物及紫色麝香葡萄果漿萃取物醱酵以移除經萃取之糖。在一實施例中，在萃取青銅色麝香葡萄果漿及紫色麝香葡萄果漿之後，但在將青銅色麝香葡萄果漿萃取物與紫色麝香葡萄果漿萃取物組合以產生所揭示之麝香葡萄果漿萃取物之前，執行醱酵。在其他實施例中，在將青銅色麝香葡萄果漿萃取物與紫色麝香葡萄果漿萃取物以所需萃取後比率(諸如約2:25/1之青銅色與紫色的比率)組合之後，執行醱酵。In some embodiments, a method of preparing a muscadine pulp extract can include separately preparing a bronze muscadine pulp extract and a purple muscadine pulp extract. For example, bronze muscadine pulp and purple musk grape pulp are extracted with water (preferably hot water). The preparation of the bronze muscadine pulp extract and the purple muscadine pulp extract may further comprise fermenting the bronze muscadine pulp extract and the purple muscadine pulp extract to remove the extracted sugar. In one embodiment, after extracting the bronze muscadine pulp and the purple muscadine pulp, but combining the bronze muscadine pulp extract with the purple muscadine pulp extract to produce the disclosed muscadine Fermentation is carried out before the pulp extract. In other embodiments, after combining the bronze muscadine pulp extract with the purple muscadine pulp extract at a desired post-extraction ratio (such as a ratio of bronze to purple of about 2:25/1), Fermented.

可藉由熟習此項技術者已知之任何方法(包括本文所述者)來執行醱酵。舉例而言，醱酵可包括將酵母及酵母營養物添加至果漿中且使萃取物醱酵直至殘餘糖含量轉化為乙醇。典型地，每1000加侖1×(未經濃縮)萃取物使用2磅酵母。在該實施例中，醱酵典型地在3天後完成。在其他實施例中，酵母之量及/或菌株及醱酵之持續時間與溫度可根據熟習此項技術者已知之個別方法而變化。在一些實施例中，可使用酶使殘餘物澄清及/或沈降或改良果漿萃取物之萃取產率。舉例而言，此等酶可包括果膠酶或來自黑麯黴(Aspergillus niger )之酶摻合物且可購自諸如史考特實驗室(Scott Laboratories)之來源。可在醱酵之前或期間將此等酶添加至果漿萃取物中。Fermentation can be carried out by any method known to those skilled in the art, including those described herein. For example, fermentation may include adding yeast and yeast nutrients to the pulp and fermenting the extract until the residual sugar content is converted to ethanol. Typically, 2 pounds of yeast is used per 1000 gallons of 1 x (unconcentrated) extract. In this embodiment, the fermentation is typically completed after 3 days. In other embodiments, the amount of yeast and/or the duration and temperature of the strain and fermentation may vary according to individual methods known to those skilled in the art. In some embodiments, the enzyme may be used to clarify and/or precipitate the residue or to improve the extraction yield of the pulp extract. For example, such enzymes may include pectinase or an enzyme blend from Aspergillus niger and may be purchased from sources such as Scott Laboratories. These enzymes can be added to the pulp extract before or during the fermentation.

在一些具體實例中，製備青銅色麝香葡萄果漿萃取物及紫色麝香葡萄果漿萃取物包括過濾青銅色麝香葡萄果漿萃取物及紫色麝香葡萄果漿萃取物。舉例而言，可在醱酵之前及/或之後過濾青銅色麝香葡萄果漿萃取物及紫色麝香葡萄果漿萃取物。可根據熟習此項技術者已知之通用方法(包括本文所揭示之過濾方法)來執行過濾。在一特定實施例中，經由適當篩孔尺寸之篩，諸如USP網(典型地為120目)或類似布濾器(諸如可購自Millipore公司之過濾器)過濾萃取物。In some embodiments, the preparation of the bronze muscadine pulp extract and the purple muscadine pulp extract comprises filtering the bronze muscadine pulp extract and the purple muscadine pulp extract. For example, bronze muscadine pulp extract and purple musk grape pulp extract can be filtered before and/or after fermentation. Filtration can be performed according to general methods known to those skilled in the art, including the filtration methods disclosed herein. In a particular embodiment, the extract is filtered through a screen of suitable mesh size, such as a USP mesh (typically 120 mesh) or a similar cloth filter (such as a filter available from Millipore Corporation).

在其他具體實例中，製備麝香葡萄果漿萃取物可包括將青銅色麝香葡萄果漿及紫色麝香葡萄果漿以所需比率組合且同時萃取青銅色麝香葡萄果漿及紫色麝香葡萄果漿。舉例而言，同時用水(較佳為熱水)萃取青銅色麝香葡萄果漿及紫色麝香葡萄果漿以產生所揭示之麝香葡萄果漿萃取物。在一些具體實例中，此方法可進一步包括使麝香葡萄果漿萃取物醱酵以移除經萃取之糖。如先前所述，可使用熟習此項技術者已知之任何醱酵方法，包括本文所述者。舉例而言，醱酵可包括將酵母及酵母營養物添加至果漿中且使萃取物醱酵直至殘餘糖含量轉化為乙醇。在一實施例中，醱酵包括每1000加侖1×萃取物添加2磅酵母且使萃取物保持於室溫下約3天或直至殘餘糖含量轉化為乙醇。此所揭示之方法亦可包括過濾麝香葡萄果漿萃取物。舉例而言，可在醱酵之前及/或之後過濾麝香葡萄果漿萃取物。可根據熟習此項技術者已知之通用方法(包括本文所揭示之過濾方法)來執行過濾。In other embodiments, preparing the muscadine pulp extract may comprise combining the bronze muscadine pulp and the purple muscadine pulp in the desired ratio and simultaneously extracting the bronze muscadine pulp and the purple muscadine pulp. For example, the bronze muscadine pulp and the purple muscadine pulp are simultaneously extracted with water (preferably hot water) to produce the disclosed muscadine pulp extract. In some embodiments, the method can further comprise fermenting the muscadine pulp extract to remove the extracted sugar. Any of the methods of fermentation known to those skilled in the art, including those described herein, can be used as previously described. For example, fermentation may include adding yeast and yeast nutrients to the pulp and fermenting the extract until the residual sugar content is converted to ethanol. In one embodiment, the fermentation comprises adding 2 pounds of yeast per 1000 gallons of 1X extract and maintaining the extract at room temperature for about 3 days or until the residual sugar content is converted to ethanol. The disclosed method can also include filtering the muscadine pulp extract. For example, the muscadine pulp extract can be filtered before and/or after fermentation. Filtration can be performed according to general methods known to those skilled in the art, including the filtration methods disclosed herein.

在某些具體實例中，製備麝香葡萄果漿萃取物之方法進一步包括濃縮青銅色麝香葡萄果漿萃取物及紫色麝香葡萄果漿萃取物以使各萃取物包括於液體中之20%至50%固體，諸如至少25%、至少30%、至少35%、至少37%、至少40%、至少42%、至少44%、至少46%或至少48%固體。在一特定實施例中，濃縮萃取物以使各萃取物包括於液體中之約40%固體。通常可使用濃縮樣品之已知方法(包括本文所揭示之濃縮樣品的方法)來濃縮青銅色及紫色萃取物。In some embodiments, the method of preparing the muscadine pulp extract further comprises concentrating the bronze muscadine pulp extract and the purple muscadine pulp extract such that each extract is included in the liquid from 20% to 50% A solid such as at least 25%, at least 30%, at least 35%, at least 37%, at least 40%, at least 42%, at least 44%, at least 46% or at least 48% solids. In a particular embodiment, the extract is concentrated such that each extract comprises about 40% solids in the liquid. The bronze and purple extracts can typically be concentrated using known methods of concentrating the sample, including the methods of concentrating the samples disclosed herein.

在一特定實施例中，為了製備具有40%固體之麝香葡萄果漿萃取物，將麝香葡萄果漿萃取物乾燥為粉末形式且以40%固體含量用水復原。或者，更可接受之商業方法為藉由經由於真空下蒸發移除萃取溶劑來濃縮。此可經由分批或連續方法達成。分批方法包括將萃取物置於容器中，抽20-29''汞柱之真空，同時加熱該容器套管以提供能量來提高溶劑之蒸氣壓。使溶劑蒸氣在容器外冷凝且冷凝速率控制冷凝物之溫度。相同原理適用於連續蒸發方法，但優點在於冷凝物曝露於高溫之時段縮短。兩種方法皆適用於濃縮本文所述之麝香葡萄果漿萃取物。In a particular embodiment, to prepare a 40% solids muscadine pulp extract, the muscadine pulp extract is dried to a powder form and reconstituted with 40% solids water. Alternatively, a more acceptable commercial method is to concentrate by removing the extraction solvent by evaporation under vacuum. This can be achieved via a batch or continuous process. The batch process involves placing the extract in a container and drawing a vacuum of 20-29 ''Hg) while heating the cannula to provide energy to increase the vapor pressure of the solvent. The solvent vapor is allowed to condense outside the vessel and the rate of condensation controls the temperature of the condensate. The same principle applies to the continuous evaporation process, but has the advantage that the time during which the condensate is exposed to high temperatures is shortened. Both methods are suitable for concentrating the muscadine pulp extract described herein.

在特定具體實例中，製備麝香葡萄果漿萃取物之方法包括分別萃取青銅色及紫色果漿，接著過濾各萃取物，隨後將青銅色與紫色果漿萃取物以所需比率組合。在一些實施例中，此方法可進一步包括使經組合之麝香葡萄果漿萃取物醱酵以移除經萃取之糖。如先前所述，可使用熟習此項技術者已知之任何醱酵方法，包括本文所述者。在一實施例中，方法包括一個以上過濾步驟，諸如在醱酵之前及之後過濾萃取物。在一些實施例中，方法可進一步包括如本文中所述濃縮萃取物。舉例而言，方法可包括藉由經由於真空下蒸發移除萃取溶劑來濃縮萃取物。In a particular embodiment, the method of preparing the muscadine pulp extract comprises separately extracting the bronze and purple pulp, followed by filtering the extracts, followed by combining the bronze and purple pulp extracts in the desired ratio. In some embodiments, the method can further comprise fermenting the combined muscadine pulp extract to remove the extracted sugar. Any of the methods of fermentation known to those skilled in the art, including those described herein, can be used as previously described. In one embodiment, the method includes more than one filtration step, such as filtering the extract before and after fermentation. In some embodiments, the method can further comprise concentrating the extract as described herein. For example, the method can include concentrating the extract by removing the extraction solvent by evaporation under vacuum.

C.具有抗氧化劑活性之組成物C. Compositions having antioxidant activity

本文揭示具有改良的抗氧化劑活性的組成物。在一些所揭示之具體實例中，該等組成物包括具有至少2%之多酚含量的麝香葡萄(圓葉葡萄)果漿萃取物及來自非麝香葡萄來源(諸如日本蓼根萃取物)之最小純度為至少5%的反式白藜蘆醇，其中麝香葡萄多酚與反式白藜蘆醇之比率在0.1/1至10/1(重量/重量)之範圍內，由此提供具有抗氧化劑活性之組成物。Compositions having improved antioxidant activity are disclosed herein. In some disclosed embodiments, the compositions include muscadine (musk) pulp extract having a polyphenol content of at least 2% and a minimum of non-musk source (such as Japanese radix extract) Trans resveratrol having a purity of at least 5%, wherein the ratio of muscadine polyphenol to trans-resveratrol is in the range of 0.1/1 to 10/1 (weight/weight), thereby providing an antioxidant Active composition.

在一些實施例中，組成物包括具有至少3%、至少3.5%、至少4%、至少4.5%、至少5%、至少6%、至少8%、至少10%、至少12%或至少14%之總多酚含量的麝香葡萄(圓葉葡萄)果漿萃取物及來自非麝香葡萄來源之最小純度為至少5%、至少10%、至少25%、至少50%、至少60%、至少70%、至少75%、至少80%、至少85%、至少90%、至少95%、至少97%或至少98%的反式白藜蘆醇。在一特定實施例中，組成物包括具有約4%之多酚含量的麝香葡萄(圓葉葡萄)果漿萃取物及來自非麝香葡萄來源之最小純度為至少98%的反式白藜蘆醇。In some embodiments, the composition comprises at least 3%, at least 3.5%, at least 4%, at least 4.5%, at least 5%, at least 6%, at least 8%, at least 10%, at least 12%, or at least 14% The total polyphenol content of the muscadine (musk) pulp extract and the minimum purity from the non-musk source is at least 5%, at least 10%, at least 25%, at least 50%, at least 60%, at least 70%, At least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97% or at least 98% trans-resveratrol. In a particular embodiment, the composition comprises a muscadine (loft grape) pulp extract having a polyphenol content of about 4% and a trans-resveratrol having a minimum purity of at least 98% from a non-musk source. .

在特定具體實例中，組成物包括具有於液體中之20%至50%固體，諸如至少23%、至少25%、至少30%、至少35%、至少37%、至少40%、至少42%、至少44%、至少46%或至少48%固體的麝香葡萄果漿萃取物。在一特定實施例中，萃取物包括於液體中之約40%固體。In a particular embodiment, the composition comprises from 20% to 50% solids in the liquid, such as at least 23%, at least 25%, at least 30%, at least 35%, at least 37%, at least 40%, at least 42%, Musk extract of muscadine at least 44%, at least 46% or at least 48% solids. In a particular embodiment, the extract comprises about 40% solids in the liquid.

在一些具體實例中，白藜蘆醇包括至少5%反式白藜蘆醇，諸如至少10%反式白藜蘆醇、至少20%反式白藜蘆醇、至少30%反式白藜蘆醇、至少40%反式白藜蘆醇、至少50%反式白藜蘆醇、至少55%、至少60%、至少70%、至少75%、至少80%、至少85%、至少90%、至少95%、至少97%或至少98%反式白藜蘆醇。在特定具體實例中，白藜蘆醇包括自日本蓼(虎杖)根萃取之至少50%反式白藜蘆醇，諸如至少55%、至少60%、至少70%、至少75%、至少80%、至少85%、至少90%、至少95%、至少97%或至少98%反式白藜蘆醇。預期所揭示之組成物中可使用其他反式白藜蘆醇來源，包括合成反式白藜蘆醇。In some embodiments, the resveratrol comprises at least 5% trans resveratrol, such as at least 10% trans resveratrol, at least 20% trans resveratrol, at least 30% trans white gourd Alcohol, at least 40% trans-resveratrol, at least 50% trans-resveratrol, at least 55%, at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, At least 95%, at least 97% or at least 98% trans-resveratrol. In a particular embodiment, the resveratrol comprises at least 50% trans-resveratrol extracted from the root of Japanese yam (polygonum cuspidatum), such as at least 55%, at least 60%, at least 70%, at least 75%, at least 80% At least 85%, at least 90%, at least 95%, at least 97% or at least 98% trans-resveratrol. Other trans-resveratrol sources, including synthetic trans-resveratrol, are contemplated for use in the disclosed compositions.

在某些具體實例中，所揭示之抗氧化劑組成物具有至少0.1/1(重量/重量)之麝香葡萄多酚與反式白藜蘆醇的比率，諸如0.25/1、0.5/1、0.6/1、0.65/1、0.7/1、0.75/1、0.8/1、0.9/1或1/1(重量/重量)。在其他具體實例中，麝香葡萄多酚與反式白藜蘆醇之比率可高達10/1(重量/重量)，諸如2/1、3/1、4/1、5/1、7.5/1或9/1(重量/重量)。在一特定實施例中，所揭示之抗氧化劑組成物具有0.75/1(重量/重量)之麝香葡萄多酚與反式白藜蘆醇的比率。In certain embodiments, the disclosed antioxidant composition has a ratio of muscadine polyphenol to trans-resveratrol of at least 0.1/1 (weight/weight), such as 0.25/1, 0.5/1, 0.6/ 1, 0.65 / 1, 0.7 / 1, 0.75 / 1, 0.8 / 1, 0.9 / 1 or 1 / 1 (weight / weight). In other embodiments, the ratio of muscadine polyphenols to trans-resveratrol can be as high as 10/1 (weight/weight), such as 2/1, 3/1, 4/1, 5/1, 7.5/1 Or 9/1 (weight/weight). In a particular embodiment, the disclosed antioxidant composition has a ratio of 0.75/1 (weight/weight) of muscadine polyphenol to trans-resveratrol.

在某些具體實例中，所揭示之抗氧化劑組成物具有0.2/1至50/1(重量/重量)範圍內之麝香葡萄果漿萃取物與反式白藜蘆醇的比率，諸如0.5/1、1/1、5/1、10/1、15/1、20/1、22/1、25/1、30/1、35/1、40/1或45/1(重量/重量)。In certain embodiments, the disclosed antioxidant composition has a ratio of muscadine pulp extract to trans-resveratrol in the range of 0.2/1 to 50/1 (weight/weight), such as 0.5/1 1, 1/1, 5/1, 10/1, 15/1, 20/1, 22/1, 25/1, 30/1, 35/1, 40/1 or 45/1 (weight/weight).

在一些具體實例中，所揭示之具有抗氧化劑活性的組成物具有每毫克多酚至少21微莫耳Trolox當量(微莫耳TE/毫克多酚)之總ORAC，諸如至少22微莫耳TE/毫克多酚、至少24微莫耳TE/毫克多酚、至少26微莫耳TE/毫克多酚、至少28微莫耳TE/毫克多酚或至少30微莫耳TE/毫克多酚。在一實施例中，所揭示之組成物具有24微莫耳TE/毫克多酚之總ORAC。In some embodiments, the disclosed composition having antioxidant activity has a total ORAC of at least 21 micromolar Trolox equivalent per microgram of polyphenol (micromolar TE per milligram of polyphenol), such as at least 22 micromoles TE/ Millions of polyphenols, at least 24 micromoles TE per milligram of polyphenols, at least 26 micromoles TE per milligram of polyphenols, at least 28 micromoles TE per milligram of polyphenols or at least 30 micromoles of TE per milligram of polyphenols. In one embodiment, the disclosed composition has a total ORAC of 24 micromoles TE per milligram of polyphenols.

在一些實施例中，所揭示之組成物可進一步包括接骨木萃取物、紫胡蘿蔔萃取物、賦形劑(諸如甘油、山梨糖醇、膠態二氧化矽或天然調味添加劑)或其組合。舉例而言，可包括接骨木萃取物及紫胡蘿蔔萃取物以提供組成物顏色或提供額外抗氧化劑活性。In some embodiments, the disclosed compositions can further include elderberry extract, purple carrot extract, excipients (such as glycerin, sorbitol, colloidal ceria or natural flavoring additives), or combinations thereof. For example, elderberry extract and purple carrot extract can be included to provide composition color or to provide additional antioxidant activity.

在一特定實施例中，組成物包括佔總組成物之23%至32%(諸如佔23%、24%、25%、26%、27%、28%、29%、30%、31%或32%)的麝香葡萄果漿萃取物(含有約4%至5%多酚含量)、佔總組成物之1.2%至3.0%(諸如佔1.3%、1.5%、1.7%、1.9%、2.1%、2.3%、2.5%、2.7%或2.9%)的反式白藜蘆醇、佔總組成物之0.12%至0.20%(諸如佔0.12%、0.14%、0.16%、0.18%或0.20%)的紫胡蘿蔔萃取物、佔總組成物之0.4%至0.6%(諸如佔0.4%、0.45%、0.5%、0.55%或0.6%)的接骨木萃取物、佔總組成物之50%至61%(諸如佔51%、52%、53%、54%、55%、56%、57%、58%、59%或60%)的山梨糖醇、佔總組成物之8%至12%(諸如佔8%、9%、10%、11%或12%)的99.7%甘油組成物及佔總組成物之1%至2%(諸如佔1.2%、1.5%、1.7%或2%)的膠態二氧化矽。在其他特定實施例中，組成物包括佔總組成物之8%至60%的麝香葡萄果漿萃取物(含有以液體重量計2%至14%麝香葡萄多酚)及佔總組成物之1.2%至3.0%的反式白藜蘆醇。In a particular embodiment, the composition comprises from 23% to 32% of the total composition (such as 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31% or 32%) muscadine pulp extract (containing about 4% to 5% polyphenol content), 1.2% to 3.0% of the total composition (such as 1.3%, 1.5%, 1.7%, 1.9%, 2.1%) , 2.3%, 2.5%, 2.7%, or 2.9%) of trans-resveratrol, from 0.12% to 0.20% of the total composition (such as 0.12%, 0.14%, 0.16%, 0.18%, or 0.20%) Purple carrot extract, from 0.4% to 0.6% of the total composition (such as 0.4%, 0.45%, 0.5%, 0.55% or 0.6%) of elderberry extract, accounting for 50% to 61% of the total composition ( Such as 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59% or 60%) of sorbitol, accounting for 8% to 12% of the total composition (such as accounted for 99.7% glycerin composition of 8%, 9%, 10%, 11% or 12%) and colloidal state of 1% to 2% (such as 1.2%, 1.5%, 1.7% or 2%) of the total composition Ceria. In other specific embodiments, the composition comprises from 8% to 60% of the total composition of the muscadine pulp extract (containing 2% to 14% muscadine polyphenols by weight of the liquid) and 1.2% of the total composition. % to 3.0% trans-resveratrol.

在一更特定實施例中，組成物包括佔總組成物之28%至30%的麝香葡萄果漿萃取物、佔總組成物之1%至2%的白藜蘆醇、佔總組成物之0.1%至0.2%的紫胡蘿蔔萃取物、佔總組成物之0.4%至0.5%的接骨木萃取物、佔總組成物之53%至56%的山梨糖醇、佔總組成物之9%至11%的99.7%甘油組成物、佔總組成物之1.5%至2%的膠態二氧化矽、佔約1%至2%之康考特葡萄萃取物、佔0.1%至0.2%之解百納葡萄萃取物及佔0.4%至0.5%之紅葡萄粉。In a more specific embodiment, the composition comprises from 28% to 30% of the total composition of the muscadine pulp extract, from 1% to 2% of the total composition of resveratrol, of the total composition. 0.1% to 0.2% purple carrot extract, 0.4% to 0.5% of the total composition of the elderberry extract, 53% to 56% of the total composition of sorbitol, 9% of the total composition to 11% of 99.7% glycerin composition, 1.5% to 2% of the total composition of colloidal cerium oxide, about 1% to 2% of Concord grape extract, accounting for 0.1% to 0.2% Nacca grape extract and 0.4% to 0.5% red grape powder.

在一甚至更特定實施例中，組成物包括佔總組成物之約28.9%的麝香葡萄果漿萃取物(含有4.1%多酚含量)、佔總組成物之約1.65%的反式白藜蘆醇、佔總組成物之約0.16%的紫胡蘿蔔萃取物、佔總組成物之約0.48%的接骨木萃取物、佔總組成物之約54.8%的山梨糖醇、佔總組成物之約10%的99.7%甘油組成物、佔總組成物之約2%的膠態二氧化矽、佔約1.27%之康考特葡萄萃取物、佔約0.16%之解百納葡萄萃取物及佔約0.48%之紅葡萄粉。In an even more specific embodiment, the composition comprises about 28.9% of the total composition of the muscadine pulp extract (containing 4.1% polyphenol content), and about 1.65% of the total composition of the trans-cigarette Alcohol, about 0.16% of the total composition of the purple carrot extract, about 0.48% of the total composition of the elderberry extract, about 54.8% of the total composition of sorbitol, about 10% of the total composition % 99.7% glycerol composition, about 2% of the total composition of colloidal cerium oxide, about 1.27% of Concord grape extract, about 0.16% of Cabernet grape extract and about 0.48 % red grape powder.

在另一特定實施例中，組成物包括佔總組成物之23%至25%的麝香葡萄果漿萃取物、佔總組成物之1%至2%的白藜蘆醇、佔總組成物之0.1%至0.2%的紫胡蘿蔔萃取物、佔總組成物之0.4%至0.5%的接骨木萃取物、佔總組成物之58%至61%的山梨糖醇、佔總組成物之9%至11%的99.7%甘油組成物、佔總組成物之1.5%至2%的膠態二氧化矽、佔約1%至2%之康考特葡萄萃取物、佔0.1%至0.2%之解百納葡萄萃取物及佔0.4%至0.5%之紅葡萄粉。In another specific embodiment, the composition comprises from 23% to 25% of the total composition of the muscadine pulp extract, from 1% to 2% of the total composition of resveratrol, of the total composition. 0.1% to 0.2% purple carrot extract, 0.4% to 0.5% of the total composition of the elderberry extract, 58% to 61% of the total composition of sorbitol, 9% of the total composition to 11% of 99.7% glycerin composition, 1.5% to 2% of the total composition of colloidal cerium oxide, about 1% to 2% of Concord grape extract, accounting for 0.1% to 0.2% Nacca grape extract and 0.4% to 0.5% red grape powder.

在另一甚至更特定實施例中，組成物包括佔總組成物之約23.77%的麝香葡萄果漿液體萃取物(含有5%多酚含量)、佔總組成物之約1.65%的反式白藜蘆醇、佔總組成物之約0.16%的紫胡蘿蔔萃取物、佔總組成物之約0.48%的接骨木萃取物、佔總組成物之約60.05%的山梨糖醇、佔總組成物之約10%的99.7%甘油組成物、佔總組成物之約2%的膠態二氧化矽、佔約1.27%之康考特葡萄萃取物、佔約0.16%之解百納葡萄萃取物及佔約0.48%之紅葡萄粉。In another even more specific embodiment, the composition comprises about 23.77% of the total composition of the muscadine pulp liquid extract (containing 5% polyphenol content), about 1.65% of the total composition of the trans white Resveratrol, about 0.16% of the total composition of the purple carrot extract, about 0.48% of the total composition of the elderberry extract, about 60.05% of the total composition of sorbitol, the total composition About 10% of 99.7% glycerin composition, about 2% of the total composition of colloidal cerium oxide, about 1.27% of Concord grape extract, about 0.16% of Cabernet grape extract and About 0.48% of red grape powder.

任一所揭示之組成物均可提供於非飲料食品、飲料、或液體或固體膳食補充劑中。在一些實施例中，以飲料形式提供所揭示之組成物。本文中之組成物(尤其食品、飲料及膳食補充劑組成物)可添加有一或多種營養物，尤其一或多種維生素及/或礦物質。維生素及礦物質之美國每日推薦攝取量(U.S. Recommended Daily Intake，USRDI)在Recommended Daily Dietary Allowance-Food and Nutrition Board,National Academy of Sciences-National Research Council中予以規定及闡明。除非本文另外規定，否則在產品中存在特定礦物質的情況下，產品包含USRDI之至少約1%(諸如至少約5%、至少約10%、至少約20%、至少約50%、至少約75%、至少約100%、至少約150%)的該礦物質。除非本文另外規定，否則在產品中存在特定維生素的情況下，產品包含USRDI之至少約1%(諸如至少約5%、至少約10%、至少約20%、至少約50%、至少約75%、至少約100%、至少約150%)的該維生素。欲添加之維生素及礦物質的量可視加工及調配物中其他成份之濃度而定。Any of the disclosed compositions can be provided in a non-beverage food, beverage, or liquid or solid dietary supplement. In some embodiments, the disclosed compositions are provided in the form of a beverage. The compositions herein (especially food, beverage, and dietary supplement compositions) may be supplemented with one or more nutrients, particularly one or more vitamins and/or minerals. U.S. Recommended Daily Intake (USRDI) is defined and clarified in the Recommended Daily Dietary Allowance-Food and Nutrition Board, National Academy of Sciences-National Research Council. Unless otherwise specified herein, in the case where a particular mineral is present in the product, the product comprises at least about 1% of USRDI (such as at least about 5%, at least about 10%, at least about 20%, at least about 50%, at least about 75). %, at least about 100%, at least about 150%) of the mineral. Unless otherwise specified herein, where a particular vitamin is present in the product, the product comprises at least about 1% of USRDI (such as at least about 5%, at least about 10%, at least about 20%, at least about 50%, at least about 75%). At least about 100%, at least about 150%) of the vitamin. The amount of vitamins and minerals to be added may depend on the concentration of the other ingredients in the processing and formulation.

該等維生素及礦物質之非限制性實例包括鐵、鋅、銅、鈣、磷、菸鹼酸、硫胺素、葉酸、泛酸、碘、維生素A、維生素C、維生素B2、維生素B3、維生素B6、維生素B12、維生素D、維生素E及維生素K。本發明之組成物中亦可包括市售來源之維生素及礦物質。Non-limiting examples of such vitamins and minerals include iron, zinc, copper, calcium, phosphorus, niacin, thiamine, folic acid, pantothenic acid, iodine, vitamin A, vitamin C, vitamin B2, vitamin B3, vitamin B6. , vitamin B12, vitamin D, vitamin E and vitamin K. Vitamins and minerals of commercial origin may also be included in the compositions of the present invention.

舉例而言，可使用適合於包含在食用產品中之此等礦物質的任何可溶性鹽，例如檸檬酸鎂、葡萄糖酸鎂、硫酸鎂、氯化鋅、硫酸鋅、碘化鉀、硫酸銅、葡萄糖酸銅及檸檬酸銅。鈣源可包括例如胺基酸螯合鈣、碳酸鈣、氧化鈣、氫氧化鈣、硫酸鈣、氯化鈣、磷酸鈣、磷酸氫鈣、磷酸二氫鈣、檸檬酸鈣、蘋果酸鈣、滴定酸鈣(calcium titrate)、葡萄糖酸鈣、calcium realate、酒石酸鈣及乳酸鈣，及尤其檸檬酸-蘋果酸鈣。如本文中所使用，「維生素A(vitamin A)」包括(但不限於)視黃醇(retinol)、β-胡蘿蔔素(β-carotene)、視黃醇棕櫚酸酯及視黃醇乙酸酯。維生素A可呈例如油、微型膠囊或囊封物之形式。亦可利用囊封抗壞血酸及可食用抗壞血酸鹽。碘源包括含碘鹽，諸如碘化鈉、碘化鉀、碘酸鉀、碘酸鈉或其混合物。此等鹽可經囊封。鐵之可接受的形式為此項技術中所熟知。併入組成物中之鐵化合物的量將視最終產物中所需之補充程度及目標消費者而廣泛變化。本揭示案之鐵強化組成物典型地含有USRDI之約5%至約100%(諸如約15%、約25%、約40%、約50%、約70%、約80%或約90%)的鐵。For example, any soluble salt suitable for such minerals contained in the edible product may be used, such as magnesium citrate, magnesium gluconate, magnesium sulfate, zinc chloride, zinc sulfate, potassium iodide, copper sulfate, copper gluconate. And copper citrate. Calcium sources may include, for example, amino acid chelated calcium, calcium carbonate, calcium oxide, calcium hydroxide, calcium sulfate, calcium chloride, calcium phosphate, calcium hydrogen phosphate, calcium dihydrogen phosphate, calcium citrate, calcium malate, titration Calcium titrate, calcium gluconate, calcium realate, calcium tartrate and calcium lactate, and especially citric acid-calcium malate. As used herein, "vitamin A" includes, but is not limited to, retinol, beta-carotene, retinyl palmitate, and retinol acetate. . Vitamin A can be in the form of, for example, an oil, a microcapsule or an encapsulate. It can also be used to encapsulate ascorbic acid and edible ascorbate. The source of iodine includes an iodide-containing salt such as sodium iodide, potassium iodide, potassium iodate, sodium iodate or a mixture thereof. These salts can be encapsulated. Acceptable forms of iron are well known in the art. The amount of iron compound incorporated into the composition will vary widely depending on the degree of replenishment desired in the final product and the target consumer. The iron-fortified compositions of the present disclosure typically comprise from about 5% to about 100% (such as about 15%, about 25%, about 40%, about 50%, about 70%, about 80%, or about 90%) of USRDI. Iron.

二價鐵典型地比三價鐵更好地被身體利用。可用於所揭示之可攝取組成物中的高度生體可利用亞鐵鹽為硫酸亞鐵、反丁烯二酸亞鐵、丁二酸亞鐵、葡萄糖酸亞鐵、乳酸亞鐵、酒石酸亞鐵、檸檬酸亞鐵、亞鐵胺基酸螯合物以及此等亞鐵鹽之混合物。雖然二價鐵典型地具有更高生體可用率，但某些鐵鹽亦可提供高度生體可利用鐵源。可用於本揭示案之食品或飲料組成物中的高度生體可利用鐵鹽為葡萄糖二酸鐵、檸檬酸鐵銨、檸檬酸鐵、硫酸鐵以及此等鐵鹽之混合物。高度生體可利用亞鐵鹽及鐵鹽之組合或混合物可用於此等可食用混合物及即食飲料中。除此等高度生體可利用亞鐵鹽及鐵鹽外，本揭示案之食品及飲料飲食組成物中亦可包括其他生體可利用鐵源。尤其適合於強化產品之其他鐵源包括某些鐵-糖-羧酸鹽複合物。Ferrous iron is typically better utilized by the body than ferrous iron. Highly fertile ferrous salts that can be used in the disclosed ingestible compositions are ferrous sulfate, ferrous fumarate, ferrous succinate, ferrous gluconate, ferrous lactate, ferrous tartrate , ferrous citrate, ferrous amine acid chelate and mixtures of such ferrous salts. While ferrous iron typically has a higher bioavailability, certain iron salts can also provide a highly bioavailable source of iron. The highly bioavailable iron salts useful in the food or beverage compositions of the present disclosure are iron digluconate, ferric ammonium citrate, ferric citrate, ferric sulfate, and mixtures of such iron salts. Highly bioavailable combinations or mixtures of ferrous and iron salts can be used in such edible mixes and ready-to-eat beverages. In addition to the use of ferrous salts and iron salts in such high biomass, the food and beverage dietary compositions of the present disclosure may also include other sources of available iron. Other sources of iron that are particularly suitable for strengthening products include certain iron-sugar-carboxylate complexes.

可用於所揭示之組成物中的鋅化合物可呈任一常用形式，諸如硫酸鋅、氯化鋅、乙酸鋅、葡萄糖酸鋅、抗壞血酸鋅、檸檬酸鋅、天冬胺酸鋅、吡啶甲酸鋅、胺基酸螯合鋅及氧化鋅。The zinc compound which can be used in the disclosed composition can be in any conventional form such as zinc sulfate, zinc chloride, zinc acetate, zinc gluconate, zinc ascorbate, zinc citrate, zinc aspartate, zinc picolinate, Amino acid chelated zinc and zinc oxide.

在一些實施例中，食品及飲料組成物亦可包括一或多種膳食纖維。「膳食纖維(dietary fiber)」意謂抗哺乳動物酶消化之複合碳水化合物，諸如植物細胞壁及海藻中所發現之碳水化合物，及由微生物醱酵所產生者。此等複合碳水化合物之實例為麩皮、纖維素、半纖維素、果膠、樹膠及黏質、海藻萃取物及生物合成樹膠。纖維素纖維之來源包括植物、果實、種子、穀物及人造纖維(例如，藉由細菌合成)。亦可使用諸如經純化植物纖維素之商品纖維或纖維素粉。天然存在之纖維包括來自完整柑橘皮、柑橘內果皮、甜菜、柑橘渣及囊固體、蘋果、杏子及西瓜皮之纖維。In some embodiments, the food and beverage composition may also include one or more dietary fibers. "Dietary fiber" means a complex carbohydrate that is resistant to enzymatic digestion by mammals, such as those found in plant cell walls and algae, and those produced by microbial fermentation. Examples of such complex carbohydrates are bran, cellulose, hemicellulose, pectin, gums and viscous, seaweed extracts and biosynthetic gums. Sources of cellulosic fibers include plants, fruits, seeds, grains, and rayon (eg, by bacterial synthesis). Commercial fibers such as purified plant cellulose or cellulose powder can also be used. Naturally occurring fibers include fibers from whole citrus peel, citrus endocarp, sugar beet, citrus pulp and sac solids, apple, apricot and watermelon rind.

可藉由已知及習知方法(例如，使用食品級酸緩衝液)將飲料酸度調整及維持在必需範圍內。典型地，在上述範圍內之飲料酸度為用於微生物抑制之最大酸度與用於所需飲料風味之最佳酸度之間的平衡。在一些實施例中，飲料組成物具有約2至約8(諸如約2至約4.5或約2.7至約4.2)之pH值。The acidity of the beverage can be adjusted and maintained within the necessary range by known and conventional methods (e.g., using a food grade acid buffer). Typically, the acidity of the beverage within the above range is the balance between the maximum acidity for microbial inhibition and the optimum acidity for the desired beverage flavor. In some embodiments, the beverage composition has a pH of from about 2 to about 8, such as from about 2 to about 4.5 or from about 2.7 to about 4.2.

有機以及無機可食用酸可用於調整飲料組成物之pH值。酸可以其未解離形式或者以其各別鹽(例如，磷酸氫二鉀或磷酸氫二鈉、磷酸二氫鉀或磷酸二氫鈉鹽)形式存在。在一些實施例中，酸包括檸檬酸、蘋果酸、反丁烯二酸、己二酸、磷酸、葡萄糖酸、酒石酸、抗壞血酸、乙酸、磷酸、丙酮酸或其混合物。酸化劑亦可用作抗氧化劑來使飲料組份穩定。常用抗氧化劑之實例包括(但不限於)抗壞血酸、乙二胺四乙酸(ethylenediaminetetraacetic acid，EDTA)及其鹽。Organic and inorganic edible acids can be used to adjust the pH of the beverage composition. The acid may be present in its undissociated form or as its individual salt (for example, dipotassium hydrogen phosphate or disodium hydrogen phosphate, potassium dihydrogen phosphate or sodium dihydrogen phosphate). In some embodiments, the acid comprises citric acid, malic acid, fumaric acid, adipic acid, phosphoric acid, gluconic acid, tartaric acid, ascorbic acid, acetic acid, phosphoric acid, pyruvic acid, or mixtures thereof. The acidulant can also be used as an antioxidant to stabilize the beverage component. Examples of commonly used antioxidants include, but are not limited to, ascorbic acid, ethylenediaminetetraacetic acid (EDTA), and salts thereof.

D.　製備具有抗氧化劑活性之麝香葡萄組成物的方法D. Method of preparing a muscat grape composition having antioxidant activity

本文亦揭示製備具有抗氧化劑活性之麝香葡萄果漿萃取物組成物的方法。本發明之組成物可根據一般技術者熟知之方法來製備。一般而言，可藉由將所有組份單獨或以合適組合形式一起溶解、分散或以其他方式混合且適當時溶解、分散或以其他方式混合於水中，用機械攪拌器攪拌直至所有成份已溶解或適當地分散來製備組成物。適當時，可組合所有單獨溶液。在需要存放穩定之組成物之情況下，可視情況在適當加工條件下對最終混合物進行巴氏殺菌或無菌填充。Also disclosed herein is a method of preparing a muscadine pulp extract composition having antioxidant activity. The compositions of the present invention can be prepared according to methods well known to those skilled in the art. In general, all components may be dissolved, dispersed or otherwise mixed, separately or in a suitable combination, and dissolved, dispersed or otherwise mixed in water, and stirred with a mechanical stirrer until all ingredients have dissolved. Or suitably dispersed to prepare a composition. All individual solutions can be combined as appropriate. The final mixture may be pasteurized or aseptically filled under appropriate processing conditions, where storage of the stable composition is desired.

此等方法可包括將具有至少2%之多酚含量的麝香葡萄(圓葉葡萄)果漿萃取物與最小純度為至少5%(諸如至少10%、至少20%、至少30%、至少40%、至少50%、至少60%、至少70%、至少80%、至少90%、至少95%、至少98%或至少99%)之來自非麝香葡萄來源的白藜蘆醇組合，其中麝香葡萄多酚與反式白藜蘆醇之比率在0.1/1至10/1(重量/重量)之範圍內，由此產生具有抗氧化劑活性之麝香葡萄果漿萃取物與反式白藜蘆醇的混合物。所揭示之方法可進一步包括製備麝香葡萄果漿萃取物，隨後將麝香葡萄果漿萃取物與反式白藜蘆醇組合。Such methods can include adding a muscadine (circle grape) pulp extract having a polyphenol content of at least 2% with a minimum purity of at least 5% (such as at least 10%, at least 20%, at least 30%, at least 40%) At least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 98% or at least 99%) of a combination of resveratrol derived from a non-musk source, wherein the muscadine is more The ratio of phenol to trans-resveratrol is in the range of 0.1/1 to 10/1 (weight/weight), thereby producing a mixture of muscadine pulp extract and trans-resveratrol having antioxidant activity. . The disclosed method can further comprise preparing a muscadine pulp extract, followed by combining the muscadine pulp extract with trans-resveratrol.

在一特定實施例中，製備麝香葡萄果漿萃取物包括將青銅色麝香葡萄果漿萃取物與紫色麝香葡萄果漿萃取物組合。在其他實施例中，製備麝香葡萄果漿萃取物包括同時萃取青銅色麝香葡萄果漿與紫色麝香葡萄果漿之混合物。在某些實施例中，根據本文中所揭示之方法製備所揭示之組成物中所包括的麝香葡萄果漿萃取物。然而，預期麝香葡萄果漿萃取物可根據此項技術中之已知方法，包括經由對萃取物進行壓碎、壓榨、萃取、過濾(若干次)及藉由真空蒸發進行濃縮、隨後冷凍來製備。在一實施例中，萃取過程僅利用水且不將例如溶劑、載劑或防腐劑之其他組份添加至萃取物自身中。該過程在保留多酚系化合物、同時減少其他分子(例如大部分糖)之存在的條件下執行。在其他實施例中，萃取過程可利用乙醇或乙醇與水之混合物。在特定實施例中，萃取過程可進一步包括使用酶來進行澄清或促進萃取。舉例而言，來自黑麯黴之酶摻合物或果膠酶可用於此等目的。商業實例包括來自史考特實驗室(Scott Laboratories)之Scottzyme KS及ScottzymePEC5L。In a particular embodiment, preparing the muscadine pulp extract comprises combining the bronze muscadine pulp extract with the purple muscadine pulp extract. In other embodiments, preparing the muscadine pulp extract comprises simultaneously extracting a mixture of bronze muscadine pulp and purple muscadine pulp. In certain embodiments, the muscadine pulp extracts included in the disclosed compositions are prepared according to the methods disclosed herein. However, it is expected that the muscadine pulp extract can be prepared according to methods known in the art, including by crushing, pressing, extracting, filtering (several times) the extract, and concentrating by vacuum evaporation followed by freezing. . In one embodiment, the extraction process utilizes only water and does not add, for example, solvents, carriers, or other components of the preservative to the extract itself. This process is carried out under conditions that retain the polyphenolic compound while reducing the presence of other molecules, such as most sugars. In other embodiments, the extraction process may utilize ethanol or a mixture of ethanol and water. In a particular embodiment, the extraction process can further comprise using an enzyme to clarify or facilitate extraction. For example, an enzyme blend or a pectinase from Aspergillus niger can be used for such purposes. Commercial examples include Scottzyme KS and Scottzyme PEC5L from Scott Laboratories.

在其他實施例中，藉由利用總酚濃度為至少2%之麝香葡萄果漿萃取物來製備所揭示之組成物。舉例而言，可使用總酚濃度為至少2%之市售麝香葡萄萃取物來製備具有抗氧化劑活性之所揭示組成物。In other embodiments, the disclosed compositions are prepared by utilizing a muscadine pulp extract having a total phenol concentration of at least 2%. For example, commercially available muscadine extracts having a total phenol concentration of at least 2% can be used to prepare the disclosed compositions having antioxidant activity.

在一特定實施例中，製備具有抗氧化劑活性之組成物的方法可進一步包括將麝香葡萄果漿萃取物及白藜蘆醇與接骨木萃取物、紫胡蘿蔔萃取物、賦形劑(諸如甘油、山梨糖醇、膠態二氧化矽或天然調味添加劑)或其組合加以組合。在一特定實施例中，方法包括將佔總組成物之23%至32%的麝香葡萄果漿萃取物(含有約4%至約5%多酚含量)與佔總組成物之1.2%至3.0%的白藜蘆醇、佔總組成物之0.12%至0.20%的紫胡蘿蔔萃取物、佔總組成物之0.4%至0.6%的接骨木萃取物、佔總組成物之50%至61%的山梨糖醇、佔總組成物之8%至12%的99.7%甘油組成物及佔總組成物之1%至2%的膠態二氧化矽組合。In a particular embodiment, the method of preparing a composition having antioxidant activity may further comprise extracting the muscadine pulp extract and resveratrol with elderberry extract, purple carrot extract, excipients (such as glycerin, Sorbitol, colloidal cerium oxide or natural flavoring additives) or combinations thereof. In a particular embodiment, the method comprises from 23% to 32% of the total composition of the muscadine pulp extract (containing from about 4% to about 5% polyphenol content) and from 1.2% to 3.0 of the total composition. % of resveratrol, 0.12% to 0.20% of total composition of purple carrot extract, 0.4% to 0.6% of total composition of elderberry extract, 50% to 61% of total composition Sorbitol, a composition of 8% to 12% of the total composition of 99.7% glycerin and a combination of 1% to 2% of the total composition of colloidal cerium oxide.

在一更特定實施例中，組成物包括將佔總組成物之約28.9%的麝香葡萄果漿萃取物(含有約4.1%多酚含量)與佔總組成物之約1.65%的白藜蘆醇、佔總組成物之約0.16%的紫胡蘿蔔萃取物、佔總組成物之約0.48%的接骨木萃取物、佔總組成物之約54.8%的山梨糖醇、佔總組成物之約10%的99.7%甘油組成物、佔總組成物之約2%的膠態二氧化矽、佔約1.27%之康考特葡萄萃取物、佔約0.16%之解百納葡萄萃取物及佔約0.48%之紅葡萄粉組合。In a more specific embodiment, the composition comprises about 28.9% of the total composition of the muscadine pulp extract (containing about 4.1% polyphenol content) and about 1.65% of the total composition of resveratrol. About 0.16% of the total composition of the purple carrot extract, about 0.48% of the total composition of the elderberry extract, about 54.8% of the total composition of sorbitol, about 10% of the total composition 99.7% glycerin composition, about 2% of the total composition of colloidal cerium oxide, about 1.27% of Concord grape extract, about 0.16% of Cabernet grape extract and about 0.48% Red grape powder combination.

在另一更特定實施例中，組成物包括將佔總組成物之約23.77%的麝香葡萄果漿液體萃取物(含有約5%多酚含量)與佔總組成物之約1.65%的反式白藜蘆醇、佔總組成物之約0.16%的紫胡蘿蔔萃取物、佔總組成物之約0.48%的接骨木萃取物、佔總組成物之約60.05%的山梨糖醇、佔總組成物之約10%的99.7%甘油組成物、佔總組成物之約2%的膠態二氧化矽、佔約1.27%之康考特葡萄萃取物、佔約0.16%之解百納葡萄萃取物及佔約0.48%之紅葡萄粉組合。In another more specific embodiment, the composition comprises about 27.67% of the total composition of the muscadine pulp liquid extract (containing about 5% polyphenol content) and about 1.65% of the total composition of the trans Resveratrol, about 0.16% of the total composition of the purple carrot extract, about 0.48% of the total composition of the elderberry extract, about 60.05% of the total composition of sorbitol, the total composition About 10% of the 99.7% glycerin composition, about 2% of the total composition of colloidal cerium oxide, about 1.27% of Concord grape extract, about 0.16% of Cabernet grape extract and Approximately 0.48% of the red grape powder combination.

在製備即飲組成物時，可視情況首先形成飲料濃縮物。一種製備飲料組成物之濃縮物形式的方法為以少於用於製備飲料組成物之所需量的水開始。另一種方法為使最終製備之飲料組成物部分脫水以僅移除一部分水及存在之任何其他揮發性液體。脫水可根據熟知程序(諸如於真空下蒸發)來完成。濃縮物可呈相對稠液體之形式。典型地藉由將合適成份(諸如電解質或乳液)添加至飲料濃縮物中來形成糖漿。接著將該糖漿與水混合形成最終飲料或最終飲料濃縮物。In the preparation of the ready-to-drink composition, the beverage concentrate may first be formed. One method of preparing a concentrate form of a beverage composition begins with less than the amount of water used to prepare the beverage composition. Another method is to partially dehydrate the final prepared beverage composition to remove only a portion of the water and any other volatile liquid present. Dehydration can be accomplished according to well known procedures such as evaporation under vacuum. The concentrate can be in the form of a relatively thick liquid. A syrup is typically formed by adding a suitable ingredient, such as an electrolyte or emulsion, to the beverage concentrate. The syrup is then mixed with water to form a final beverage or final beverage concentrate.

所揭示之組成物的基本上乾燥混合物可藉由將適當量及比率之所有所需乾燥成份摻合在一起來製備。或者，可使最終製備之飲料組成物脫水以產生飲料組成物之基本上乾燥混合物。呈例如散劑、顆粒或錠劑形式之基本上乾燥混合物可稍後溶解於適當量之碳酸或非碳酸水中以產生最終可飲用飲料或結合水服用。The substantially dry mixture of the disclosed compositions can be prepared by blending together all of the desired dry ingredients in the appropriate amounts and ratios. Alternatively, the final prepared beverage composition can be dewatered to produce a substantially dry mixture of the beverage composition. The substantially dry mixture, for example in the form of a powder, granule or lozenge, can be dissolved later in an appropriate amount of carbonated or non-carbonated water to produce the final drinkable beverage or combined with water.

作為食品形式，所揭示之組成物可調配為任何可選形式，例如顆粒態、晶粒態、糊劑態、凝膠態、固態或液態。在此等形式中，可包括各種其他物質，諸如黏合劑、崩解劑、增稠劑、分散劑、再吸收促進劑、調味劑、緩衝劑、界面活性劑、溶解助劑、防腐劑、乳化劑、等滲劑、穩定劑或pH值控制劑。在特定實施例中，當所揭示之組成物用作保持健康(包括(但不限於)改善皮膚品質)之食品、功能性食品等時，較佳以使得表明本揭示案之預定作用足以提供抗氧化劑活性的量含有本發明之組成物及萃取物(例如，麝香葡萄果漿萃取物、白藜蘆醇、紫胡蘿蔔萃取物及接骨木萃取物)之活性成份。As a food form, the disclosed compositions can be formulated in any optional form, such as particulate, crystalline, pasty, gel, solid or liquid. In such forms, various other materials may be included, such as binders, disintegrants, thickeners, dispersants, resorption enhancers, flavoring agents, buffers, surfactants, dissolution aids, preservatives, emulsifying agents. Agent, isotonicity agent, stabilizer or pH control agent. In certain embodiments, when the disclosed compositions are used as foods, functional foods, and the like that maintain health, including but not limited to improving skin quality, preferably, such that the intended effect of the present disclosure is sufficient to provide resistance The amount of oxidant activity comprises the active ingredients of the compositions and extracts of the invention (e.g., muscadine pulp extract, resveratrol, purple carrot extract, and elderberry extract).

E.　用於抑制(例如防止)細胞衰老之方法及套組E. Methods and kits for inhibiting (eg, preventing) cellular aging

本文顯示所揭示之組成物具有改良的親脂性抗氧化劑活性。已知氧化應力為造成正常衰老之主要機制。亦已知親脂性抗氧化劑能夠抑制各種類型之皮膚損傷。基於此等觀察結果，揭示抑制細胞衰老(諸如抑制或降低自由基產生或活性)之方法。在一實施例中，方法包括向有需要之個體投予足以抑制或減少一或多個與細胞衰老相關之過程之劑量的所揭示之具有抗氧化劑活性的組成物。舉例而言，抑制一或多個與細胞衰老相關之過程包括降低或抑制攝取該組成物之該個體體內的自由基形成或活性。The compositions disclosed herein are shown to have improved lipophilic antioxidant activity. Oxidative stress is known to be the primary mechanism responsible for normal aging. Lipophilic antioxidants are also known to inhibit various types of skin damage. Based on these observations, methods for inhibiting cellular senescence, such as inhibiting or reducing free radical production or activity, are disclosed. In one embodiment, the method comprises administering to a subject in need thereof a disclosed composition having antioxidant activity sufficient to inhibit or reduce one or more doses associated with cellular aging. For example, inhibiting one or more processes associated with cellular senescence includes reducing or inhibiting free radical formation or activity in the subject ingesting the composition.

在一些所揭示之具體實例中，方法包括投予包括來源於青銅色麝香葡萄果漿及紫色麝香葡萄果漿之麝香葡萄(圓葉葡萄)果漿萃取物的可攝取組成物(諸如本文所述者)。在一些具體實例中，麝香葡萄果漿萃取物中青銅色麝香葡萄果漿萃取物與紫色麝香葡萄果漿萃取物之比率在0.1至10(重量/重量)，諸如0.3至3(重量/重量)之範圍內。舉例而言，青銅色麝香葡萄果漿萃取物與紫色麝香葡萄果漿萃取物之比率為約2.75/約1(重量/重量)、2.5/約1(重量/重量)、約2.25/約1(重量/重量)、約2/約1(重量/重量)、約1.5/約1(重量/重量)或約1/約1(重量/重量)。如本文中所使用，術語「約」係定義為±0.5。在一特定實施例中，青銅色麝香葡萄果漿萃取物與紫色麝香葡萄果漿萃取物之比率為約2.25/約1(重量/重量)。In some disclosed embodiments, the method comprises administering an ingestible composition (such as described herein) comprising a muscadine (circle grape) pulp extract derived from bronze muscadine pulp and purple muscadine pulp. By). In some embodiments, the ratio of the bronze muscadine pulp extract to the purple muscadine pulp extract in the muscadine pulp extract is from 0.1 to 10 (weight/weight), such as from 0.3 to 3 (weight/weight). Within the scope. For example, the ratio of bronze muscadine pulp extract to purple muscadine pulp extract is about 2.75 / about 1 (weight / weight), 2.5 / about 1 (weight / weight), about 2.25 / about 1 ( Weight/weight), about 2/about 1 (weight/weight), about 1.5/about 1 (weight/weight) or about 1/about 1 (weight/weight). As used herein, the term "about" is defined as ±0.5. In a particular embodiment, the ratio of bronze muscadine pulp extract to purple muscadine pulp extract is about 2.25 / about 1 (weight / weight).

在某些具體實例中，經口投予之麝香葡萄(圓葉葡萄)果漿萃取物具有至少2%之多酚含量。舉例而言，多酚含量為至少3%、至少3.5%、至少4%、至少4.5%、至少5%、至少6%、至少8%、至少10%、至少12%或至少14%。在一特定實施例中，麝香葡萄(圓葉葡萄)果漿萃取物具有約4%之多酚含量。In some embodiments, the orally administered Muscat (Clover) pulp extract has a polyphenol content of at least 2%. For example, the polyphenol content is at least 3%, at least 3.5%, at least 4%, at least 4.5%, at least 5%, at least 6%, at least 8%, at least 10%, at least 12%, or at least 14%. In a particular embodiment, the muscadine (circle grape) pulp extract has a polyphenol content of about 4%.

在一些具體實例中，方法包括投予包括麝香葡萄果漿萃取物以及其他額外成份(包括(但不限於)白藜蘆醇)之可攝取組成物。在一些實施例中，方法包括投予具有約0.75/1(重量/重量)之麝香葡萄多酚與白藜蘆醇的比率之口服組成物。在其他實施例中，方法包括投予至少包括佔總組成物之8%至60%的麝香葡萄果漿萃取物(含有以液體重量計2%至14%，諸如4%、6%、8%、10%或12%麝香葡萄多酚)及佔總組成物之1.2%至3.0%的反式白藜蘆醇的組成物。In some embodiments, the method comprises administering an ingestible composition comprising a muscadine pulp extract and other additional ingredients including, but not limited to, resveratrol. In some embodiments, the method comprises administering an oral composition having a ratio of muscadine polyphenol to resveratrol of about 0.75/1 (weight/weight). In other embodiments, the method comprises administering at least 8% to 60% of the total composition of the muscadine pulp extract (containing 2% to 14% by weight of the liquid, such as 4%, 6%, 8%) , 10% or 12% muscadine polyphenols) and a composition of trans-resveratrol from 1.2% to 3.0% of the total composition.

在另一具體實例中，方法包括投予包括具有至少2%之多酚含量的麝香葡萄果漿萃取物及來自非麝香葡萄來源(諸如日本蓼根萃取物)之最小純度為至少5%的白藜蘆醇之口服組成物，其中麝香葡萄多酚與反式白藜蘆醇之比率在0.1/1至10/1(重量/重量)之範圍內，由此提供具有足以抑制或減少一或多個與細胞衰老相關之過程(包括(但不限於)改善皮膚品質)的抗氧化劑活性之組成物。在一特定實施例中，方法包括投予包括具有至少2%之多酚含量的麝香葡萄(圓葉葡萄)果漿萃取物及來自非麝香葡萄來源(諸如日本蓼根萃取物)之最小純度為至少5%的白藜蘆醇之組成物，其中麝香葡萄果漿萃取物與白藜蘆醇之比率在0.2/1至50/1(重量/重量)，諸如5/1至50/1(重量/重量)或20/1至50/1(重量/重量)之範圍內，由此提供具有抗氧化劑活性之組成物。在一些實施例中，組成物具有每毫克多酚至少24微莫耳Trolox當量(微莫耳TE/毫克多酚)之總ORAC。In another embodiment, the method comprises administering a muscadine pulp extract comprising a polyphenol content of at least 2% and a white color having a minimum purity of at least 5% from a non-musk source (such as Japanese radix root extract) An oral composition of resveratrol wherein the ratio of muscadine polyphenol to trans-resveratrol is in the range of from 0.1/1 to 10/1 (weight/weight), thereby providing sufficient to inhibit or reduce one or more A composition of antioxidant activity associated with cellular aging, including but not limited to improving skin quality. In a particular embodiment, the method comprises administering a muscadine (musk) pulp extract comprising at least 2% polyphenol content and a minimum purity from a non-musk source (such as Japanese radix root extract). At least 5% of the composition of resveratrol, wherein the ratio of the muscadine pulp extract to resveratrol is from 0.2/1 to 50/1 (weight/weight), such as from 5/1 to 50/1 (weight) /weight) or in the range of 20/1 to 50/1 (weight/weight), thereby providing a composition having antioxidant activity. In some embodiments, the composition has a total ORAC of at least 24 micromoles of Trolox equivalent per microgram of polyphenol (micromolar TE per milligram of polyphenol).

在其他具體實例中，該等方法藉由經由塗覆含有單獨麝香葡萄果漿萃取物或其與其他活性成份(包括(但不限於)白藜蘆醇)之組合的溶液或局部軟膏來抑制或降低皮膚細胞中之自由基形成或活性而改善皮膚品質。在一實施例中，溶液或局部軟膏包括所揭示之麝香葡萄果漿萃取物而無白藜蘆醇。在另一實施例中，溶液或局部軟膏包括所揭示之麝香葡萄果漿萃取物與白藜蘆醇兩者，諸如包括麝香葡萄多酚濃縮物之麝香葡萄果漿萃取物以使麝香葡萄多酚與白藜蘆醇之比率為約0.75/1(重量/重量)。該方法可藉由臨床醫師或其他保健提供者來執行。該方法亦經設計供家庭使用。該方法可減少與衰老相關之皮膚變化的出現，明顯減少人類皮膚皺紋，且改善皮膚之組織品質。亦提供用於改善皮膚品質之組成物及套組，其可包括所揭示之具有抗氧化劑活性的組成物(包括口服或局部)或麝香葡萄萃取物及一或多種其他抗衰老組成物(諸如一或多種其他抗氧化劑)。In other embodiments, the methods are inhibited by coating a solution or topical ointment containing a separate muscadine pulp extract or a combination thereof with other active ingredients including, but not limited to, resveratrol. Reduces free radical formation or activity in skin cells to improve skin quality. In one embodiment, the solution or topical ointment comprises the disclosed muscadine pulp extract without resveratrol. In another embodiment, the solution or topical ointment comprises both the disclosed muscadine pulp extract and resveratrol, such as muscadine pulp extract comprising muscadine polyphenol concentrate to make muscadine polyphenols The ratio to resveratrol is about 0.75/1 (weight/weight). The method can be performed by a clinician or other healthcare provider. The method is also designed for home use. This method can reduce the occurrence of skin changes associated with aging, significantly reduce wrinkles in human skin, and improve the tissue quality of the skin. Compositions and kits for improving skin quality are also provided, which may include the disclosed compositions having antioxidant activity (including oral or topical) or muscadine extract and one or more other anti-aging compositions (such as a Or a variety of other antioxidants).

任何皮膚表面(例如皮膚之表皮)均可藉由使用本文提供之方法來治療。可治療之皮膚表面包括(但不限於)眶骨膜、唇、面頰、鼻唇溝、前額、頸部、上唇紋、胃、頸部、背部、胸部、手、腿、腳或其任何組合。在一實施例中，任何面部表面之皮膚可使用本文所提供之方法來治療。該方法可應用於任何面部區域及/或任何體表區域，其他直接應用區域為胸部及頸部。可在同一治療時期內治療一個以上皮膚表面。在特定實施例中，將所揭示之組成物或萃取物的液體或乳膏形式實質上均勻地塗覆於皮膚表面上，於皮膚上形成一層組成物或萃取物。Any skin surface, such as the epidermis of the skin, can be treated by using the methods provided herein. The treatable skin surface includes, but is not limited to, the periosteum, the lips, the cheeks, the nasolabial folds, the forehead, the neck, the upper lip, the stomach, the neck, the back, the chest, the hands, the legs, the feet, or any combination thereof. In one embodiment, the skin of any facial surface can be treated using the methods provided herein. The method can be applied to any facial area and/or any body surface area, and other direct application areas are the chest and neck. More than one skin surface can be treated during the same treatment period. In a particular embodiment, the liquid or cream form of the disclosed composition or extract is applied substantially uniformly to the surface of the skin to form a layer of composition or extract on the skin.

所揭示之組成物或萃取物可含有通常用於化妝品中之其他物質，例如：香料；抗微生物劑；抗菌劑；加脂劑；錯合劑及錯隔劑；珠光劑；植物萃取物；維生素，諸如視黃醇或維生素C；活性劑；防腐劑；殺菌劑；界面活性劑、染料、著色劑、色素或具有著色作用之任何物質；乳化劑；增稠劑；軟化物質、增濕物質及/或保濕物質；或美容或皮膚學調配物之其他常見組份，諸如醇、多元醇、聚合物、泡沫穩定劑、電解質、有機溶劑或聚矽氧衍生物。組成物亦可包含功能性添加劑，諸如去角質劑、氧化劑、防曬劑及皮膚光滑劑。組成物亦可含有認為有益於皮膚及下層皮下組織之中胚層療法(mesotherapy)注射治療的組份，包括抗氧化劑，諸如二甲基胺基乙醇、α硫辛酸及抗壞血酸。組成物中亦可包括可增強組成物或萃取物之功效的藥劑，諸如親脂性及疏脂性滲透增強劑(例如，檸檬酸三乙酯、丙二醇、脂肪酸酯及其他物質)。組成物中亦可包括可增強皮膚之血管灌注的藥劑，諸如胺茶鹼(aminophyllin)或己酮可可鹼(pentoxifylline)。組成物中亦可包括可增強皮膚之膨壓及張力以及使得下層皮下組織結構收縮或皺縮的藥劑，諸如磷脂醯膽鹼(phosphatidyl choline)及去氧膽酸硫酸鹽(deoxycholate sulfate)。組成物中亦可包括可提供激素益處之生理物質，諸如為皮膚之***性(estrogenic)或睾固酮性(testosterrogenic)刺激物的物質，包括雌三醇(estriol)及睾固酮(testosterone)。The disclosed compositions or extracts may contain other substances commonly used in cosmetics, such as: perfumes; antimicrobial agents; antibacterial agents; fatliquors; complexing agents and spacers; pearlizing agents; plant extracts; Such as retinol or vitamin C; active agent; preservative; fungicide; surfactant, dye, colorant, pigment or any substance with coloring; emulsifier; thickener; softening substance, moisturizing substance and / Or a moisturizing substance; or other common components of a cosmetic or dermatological formulation, such as an alcohol, a polyol, a polymer, a foam stabilizer, an electrolyte, an organic solvent or a polyoxyl derivative. The composition may also contain functional additives such as exfoliants, oxidizing agents, sunscreens, and skin smoothing agents. The composition may also contain components believed to be beneficial for skin and subcutaneous tissue mesotherapy injection therapy, including antioxidants such as dimethylaminoethanol, alpha lipoic acid, and ascorbic acid. Agents may also be included in the composition to enhance the efficacy of the composition or extract, such as lipophilic and lipophobic penetration enhancers (e.g., triethyl citrate, propylene glycol, fatty acid esters, and other materials). Agents that enhance vascular perfusion of the skin, such as aminophyllin or pentoxifylline, may also be included in the composition. Agents may also be included in the composition which enhance the swell and tension of the skin and cause the underlying subcutaneous tissue to shrink or shrink, such as phosphatidyl choline and deoxycholate sulfate. Physiological substances that provide hormonal benefits, such as substances that are estrogen or testosterrogenic stimuli of the skin, including estriol and testosterone, may also be included in the composition. ).

所揭示之麝香葡萄組成物可包括一或多種防腐劑。此等防腐劑包括例如Opthiphen^TM (來自International Specialty Products)、GeogardUltra(來自Lonza)、歐盟化妝品指令(European Union Cosmetic Directive)中所列之防腐劑及其他防腐劑，諸如甲醛供體(諸如DMDM乙內醯脲，其可以商品名GLYDAN自Lonza獲得)、丁基胺基甲酸碘丙酯(例如，可以商品名GLYCACIL-STM自LONZATM獲得及/或以商品名DEKABEN LMBTM自Jan Dekker獲得者)、對羥基苯甲酸酯(例如，對羥基苯甲酸烷基酯，諸如對羥基苯甲酸甲酯、對羥基苯甲酸乙酯、對羥基苯甲酸丙酯及/或對羥基苯甲酸丁酯)、苯氧基乙醇、乙醇、苯甲酸及水楊酸。防腐系統可進一步包括防腐助劑，諸如辛氧基甘油(octoxyglycerin)或大豆油(glycine soya)。其他防腐劑或防腐助劑包括二溴氰基丁烷(2-溴-2-溴甲基戊二腈)、丁基胺基甲酸3-碘-2-丙炔酯、2-溴-2-硝基丙-1,3-二醇、咪唑啶基脲、5-氯-2-甲基-4-異噻唑啉-3-酮、2-氯乙醯胺、氯苄烷銨及苄醇。The disclosed muscadine composition can include one or more preservatives. Such preservatives include, for example, Opthiphen ^TM (from International Specialty Products), Geogard Ultra (from Lonza), preservatives listed in the European Union Cosmetic Directive and other preservatives, such as formaldehyde donors (such as DMDM carbendazim, which can be traded under the trade name GLYDAN) Obtained from Lonza), iodopropyl butyl carbamate (for example, available from LONZATM under the trade name GLYCACIL-STM and/or from Jan Dekker under the trade name DEKABEN LMBTM), for example, p-hydroxybenzoate (for example, Alkyl hydroxybenzoate, such as methyl paraben, ethyl p-hydroxybenzoate, propyl p-hydroxybenzoate and/or butyl p-hydroxybenzoate, phenoxyethanol, ethanol, benzoic acid and water Salicylic acid. The preservative system may further comprise an anti-corrosion aid such as octoxyglycerin or glycine soya. Other preservatives or preservatives include dibromocyanobutane (2-bromo-2-bromomethylglutaronitrile), 3-iodo-2-propynyl butylcarbamate, 2-bromo-2- Nitropropane-1,3-diol, imidazolidinyl urea, 5-chloro-2-methyl-4-isothiazolin-3-one, 2-chloroacetamide, benzalkonium chloride and benzyl alcohol.

組成物亦可包括一或多種調節劑，諸如水溶性調節劑。其他調節劑包括例如根據國際化妝品成份命名法(International Nomenclature for Cosmetic Ingredients，INCI)稱為聚四級銨(尤其聚四級銨-1至聚四級銨-56)之化合物。The composition may also include one or more conditioning agents, such as water soluble conditioning agents. Other regulators include, for example, compounds known as polytetra-ammonium (especially polytetra-ammonium-1 to poly-quaternary ammonium-56) according to the International Nomenclature for Cosmetic Ingredients (INCI).

組成物可自軟管、廣口瓶、瓶子、泵、罐、噴霧罐或噴霧瓶或自一些其他已知容器分配。The composition can be dispensed from a hose, jar, bottle, pump, can, spray can or spray bottle or from some other known container.

本文亦揭示用於改善皮膚品質供家庭或臨床醫師使用之麝香葡萄果漿萃取物組成物及套組。在一實施例中，此套組適合於臨床醫師或美容師使用。在該實施例中，套組可進一步包括輔助塗覆組成物之塗覆器。套組通常亦包括使用說明書。此等說明書可為手寫或呈數位格式(諸如錄影帶、DVD或CD)以用於諸如電腦、CD播放機、mp3播放機或DVD播放機及其類似物之電子裝置。在另一實施例中，套組適合於家庭使用。此套組可包括用於塗覆組成物之塗覆器，諸如海綿或布。或者，可使用一或多根手指塗覆組成物。套組通常亦包括使用說明書。此等說明書可為手寫或呈數位格式(諸如錄影帶、DVD或CD)以用於諸如電腦、CD播放機、mp3播放機或DVD播放機及其類似物之電子裝置。Also disclosed herein are muscadine pulp extract compositions and kits for use in improving skin quality for use by a family or clinician. In one embodiment, the kit is suitable for use by a clinician or beautician. In this embodiment, the kit may further include an applicator that assists in coating the composition. The kit usually also includes instructions for use. Such instructions may be handwritten or in digital format (such as videotape, DVD or CD) for use with electronic devices such as computers, CD players, mp3 players or DVD players and the like. In another embodiment, the kit is suitable for home use. This kit may include an applicator for coating the composition, such as a sponge or cloth. Alternatively, the composition can be applied using one or more fingers. The kit usually also includes instructions for use. Such instructions may be handwritten or in digital format (such as videotape, DVD or CD) for use with electronic devices such as computers, CD players, mp3 players or DVD players and the like.

在一些實施例中，套組包括位於兩個單獨容器中或以單一組成物形式位於單一容器中之所揭示之組成物或萃取物及一或多種其他抗衰老化合物(諸如另一抗氧化劑(例如，維生素C、維生素E、硒及/或β-胡蘿蔔素))中的一者。在一實施例中，組成物或萃取物塗覆於目標皮膚區域。在一特定非限制性實施例中，麝香葡萄果漿萃取物抗衰老套組可由家庭人員如下使用。首先，藉由使用塗覆器(諸如海綿或布)或藉由使用一或多根手指將實測量之組成物或萃取物塗覆於皮膚表面，以於皮膚表面上提供一層均勻的組成物或萃取物。讓組成物或萃取物保留某一時間長度以被動吸收至皮膚表面中，或可藉由用指尖輕輕地將組成物摩擦至皮膚表面中來促進皮膚表面對組成物/萃取物之吸收。通常，諸如化妝塗覆及塗覆其他增濕劑之正常皮膚護理程序可在塗覆麝香葡萄果漿組成物/萃取物之後繼續進行。In some embodiments, the kit includes the disclosed composition or extract and one or more other anti-aging compounds (such as another antioxidant) (eg, in two separate containers or in a single container in a single container) One of vitamin C, vitamin E, selenium and/or beta-carotene). In one embodiment, the composition or extract is applied to the target skin area. In a specific, non-limiting embodiment, the muscadine pulp extract anti-aging kit can be used by a household member as follows. First, to provide a uniform composition on the surface of the skin by using an applicator (such as a sponge or cloth) or by applying a measured composition or extract to the surface of the skin using one or more fingers. Extracts. The composition or extract is allowed to remain passively absorbed into the surface of the skin for a certain length of time, or the absorption of the composition/extract by the skin surface can be promoted by gently rubbing the composition into the surface of the skin with a fingertip. In general, normal skin care procedures such as cosmetic coating and application of other moisturizing agents may continue after the muscadine pulp composition/extract is applied.

此過程可如所述每日執行兩次，每日、每隔一天、雙週、每週、每隔一週或每月執行，或歷時一些其他時間間隔，諸如每3至5天一次。改善皮膚品質包括逆轉與天然或先天衰老相關之皮膚變化、減緩該等皮膚變化之進展或防止該等皮膚變化。如本文中所使用，防止其變化係指以任何程度延緩皮膚變化開始。舉例而言，改善皮膚品質包括逆轉與自由基形成及活性相關之皮膚變化、減緩該等皮膚變化之進展或防止該等皮膚變化。在一實施例中，改善皮膚品質包括逆轉與曬傷相關之皮膚變化或與曝露於日光或其他形式之光化輻射(例如，UV輻射及曬黑室)相關的光衰老皮膚變化、減緩該等皮膚變化之進展或防止該等皮膚變化。作為另一實施例，改善皮膚品質亦可包括逆轉由外在因素引起之皮膚變化、減緩該等皮膚變化之進展或防止該等皮膚變化，該等外在因素包括(但不限於)輻射、空氣污染、風、寒冷、潮濕、熱、化學物質、煙霧、香煙煙熏及其組合。改善皮膚品質亦可包括逆轉、防止或減少可例如由某些皮膚病狀(例如痤瘡)、感染(例如利什曼病(leishmaniasis))或損傷(例如擦傷、刺傷、劃傷或手術創口)引起之瘢痕。對皮膚之改善亦可包括以下至少一種：使面線顯得較不明顯，使面線及/或皺紋感覺飽滿，改善眼眶下線紋及/或眼周線紋之外觀，改善魚尾紋之外觀，減少及/或減弱皺紋、尤其面頰、前額上(例如眼睛之間的垂直皺紋、眼睛上方之水平皺紋)及/或口周圍之面部皺紋、及尤其深皺紋、褶皺或皺褶之出現，改善皮膚柔度，減少及/或消除細紋及/或深紋、褶皺及皺褶，且使皮膚光滑。量測改良的皮膚品質之方法為此項技術中所已知。參見例如美國專利第6,866,856號及第6,682,763號。This process can be performed twice daily as described, daily, every other day, biweekly, weekly, every other week or monthly, or for some other time interval, such as every 3 to 5 days. Improving skin quality includes reversing skin changes associated with natural or congenital aging, slowing the progression of such skin changes or preventing such skin changes. As used herein, preventing its change refers to the beginning of delaying skin changes to any degree. For example, improving skin quality includes reversing skin changes associated with free radical formation and activity, slowing the progression of such skin changes, or preventing such skin changes. In one embodiment, improving skin quality includes reversing skin changes associated with sunburn or light aging skin changes associated with exposure to sunlight or other forms of actinic radiation (eg, UV radiation and tanning chambers), slowing down such The progression of skin changes or prevents such skin changes. As another example, improving skin quality may also include reversing skin changes caused by extrinsic factors, slowing the progression of such skin changes, or preventing such skin changes, including but not limited to radiation, air. Pollution, wind, cold, humidity, heat, chemicals, smoke, cigarette smoke, and combinations thereof. Improving skin quality may also include reversing, preventing or reducing, for example, caused by certain skin conditions (eg, acne), infections (eg, leishmaniasis), or injuries (eg, abrasions, stab wounds, scratches, or surgical wounds). Scars. The improvement to the skin may also include at least one of the following: making the upper thread less visible, making the upper thread and/or wrinkles feel full, improving the appearance of the underline and/or the contour of the eye contour, improving the appearance of the crow's feet, and reducing / / to reduce wrinkles, especially on the cheeks, forehead (such as vertical wrinkles between the eyes, horizontal wrinkles above the eyes) and / or facial wrinkles around the mouth, and especially the appearance of deep wrinkles, wrinkles or wrinkles, improve skin softness Degree, reduces and/or eliminates fine lines and/or deep lines, wrinkles and wrinkles, and smoothes the skin. Methods for measuring improved skin quality are known in the art. See, for example, U.S. Patent Nos. 6,866,856 and 6,682,763.

可藉由實施該等方法及使用本文所揭示之套組來治療的皮膚變化包括例如皺紋(包括(但不限於)人類面部皺紋)、皺褶、深皺、褶皺及細紋，皮膚線紋加深，皮膚變薄，防止或減少瘢痕，皮膚變黃，色斑，色素過多，出現色素及/或無色素老人斑，皮革樣皮質(leatheriness)，損失彈性，損失反彈性(recoilability)，損失膠原纖維，彈性纖維變化異常，真皮小血管退化，皮膚表面上形成日光增加可見之血管結構，及其組合。Skin changes that can be treated by performing such methods and using the kits disclosed herein include, for example, wrinkles (including but not limited to human facial wrinkles), wrinkles, deep wrinkles, wrinkles, and fine lines, deepening of skin lines Thinning of the skin, preventing or reducing scarring, yellowing of the skin, pigmentation, hyperpigmentation, pigmentation and/or pigment-free age spots, leatheriness, loss of elasticity, loss of recoilability, loss of collagen fibers The elastic fiber changes abnormally, the small blood vessels of the dermis degenerate, and the surface of the skin forms a vascular structure with increased sunlight, and a combination thereof.

改善皮膚品質包括減少、縮減一或多種上述皮膚變化及/或使該等皮膚變化減至最少。改善皮膚品質會使得皮膚具有更年輕之外觀。改善皮膚品質會使得皮膚具有更光滑、滋潤(較不乾燥)或較少起鱗之外觀。舉例而言，在某些具體實例中，改善皮膚品質可包括粗糙度、乾燥度或起鱗程度降低。改善皮膚品質包括淡化及改善線紋及皺紋，改善膨壓及張力，其中所觀察到之所需效果為提拉及緊致。Improving skin quality includes reducing, reducing, and/or minimizing one or more of the above-described skin changes. Improving skin quality will give the skin a younger look. Improving skin quality will result in a smoother, moisturized (less dry) or less scaly appearance. For example, in certain embodiments, improving skin quality can include a reduction in roughness, dryness, or degree of scaling. Improving skin quality includes desalination and improvement of lines and wrinkles, and improvement of swell pressure and tension. The desired effect is lift and firmness.

皮膚之組織品質可得到改善，包括軟度、柔度及平滑度，致使光澤度、透明度及亮度得到增強。可主觀上及客觀上量測之皮膚的其他及重要品質包括(但不限於)皮膚鬆弛性或相反皮膚緊致性，及皮膚中之組織細紋及粗紋的存在及程度。The texture of the skin can be improved, including softness, softness and smoothness, resulting in enhanced gloss, clarity and brightness. Other and important qualities of the subject that can be subjectively and objectively measured include, but are not limited to, skin sagging or reverse skin firmness, and the presence and extent of fine lines and streaks in the skin.

此等品質為判斷外觀之外部態樣(例如皮膚衰老)的相同品質。藉由本文所揭示之治療方法及套組改善此等品質產生基於視覺外觀判斷之益處。藉由本文所揭示之方法改變皮膚品質減少皮膚衰老之出現。These qualities are the same qualities that determine the external appearance of the appearance (eg skin aging). Improving the quality by the methods and kits disclosed herein produces benefits based on visual appearance judgments. Changing skin quality by the methods disclosed herein reduces the appearance of skin aging.

所需益處可不僅包括對皮膚之生理益處，而且包括治療性及藥理學益處，諸如可能預防及治療惡性疾病，無論藉由化學預防抑或增強光動力療法。益處亦可包括藉由包括抑制皮脂腺活性之組成物來治療及抑制痤瘡。The desired benefit may include not only physiological benefits to the skin, but also therapeutic and pharmacological benefits, such as possible prevention and treatment of malignant diseases, whether by chemoprevention or enhanced photodynamic therapy. Benefits may also include treating and inhibiting acne by including a composition that inhibits sebaceous gland activity.

F.　醫藥組成物F. Pharmaceutical composition

所揭示之麝香葡萄果漿萃取物及組成物可至少適用於抑制(例如防止)一或多個氧化過程，諸如與細胞事件(諸如細胞衰老)相關之自由基形成。因此，本文亦描述包含單獨至少一種所揭示之麝香葡萄果漿萃取物或其與白藜蘆醇之組合的醫藥組成物。The disclosed muscadine pulp extracts and compositions can be at least suitable for inhibiting (e.g., preventing) one or more oxidation processes, such as free radical formation associated with cellular events such as cellular senescence. Accordingly, a pharmaceutical composition comprising at least one of the disclosed muscadine pulp extracts alone or in combination with resveratrol is also described herein.

用於醫藥組成物之調配物為此項技術中所熟知。舉例而言，Remington’s Pharmaceutical Sciences ,E. W. Martin,Mack Publishing公司,Easton,PA,第19版,1995描述適合於醫藥傳遞單獨至少一種所揭示之麝香葡萄果漿萃取物或其與白藜蘆醇之組合的例示性調配物(及其組份)。在一些實施例中，該等組成物亦包括其他藥劑，諸如紫胡蘿蔔根萃取物或接骨木萃取物。包含至少一種此等化合物之醫藥組成物可經調配用於人類醫學或獸醫學。所揭示之醫藥組成物的特定調配物可例如視投藥模式(例如口服、局部或非經腸)及/或欲治療之病狀(例如自由基產生或活性)而定。在一些具體實例中，除諸如麝香葡萄果漿萃取物之至少一種活性成份外，調配物亦包括醫藥學上可接受之載劑。Formulations for use in pharmaceutical compositions are well known in the art. For example, Remington's Pharmaceutical Sciences , EW Martin, Mack Publishing Company, Easton, PA, 19th edition, 1995 describes suitable for the delivery of at least one of the disclosed muscadine pulp extracts alone or in combination with resveratrol. An exemplary formulation (and its components). In some embodiments, the compositions also include other agents, such as purple carrot root extract or elderberry extract. Pharmaceutical compositions comprising at least one such compound can be formulated for use in human medicine or veterinary medicine. Particular formulations of the disclosed pharmaceutical compositions may, for example, depend on the mode of administration (e.g., oral, topical or parenteral) and/or the condition to be treated (e.g., free radical production or activity). In some embodiments, the formulation also includes a pharmaceutically acceptable carrier in addition to at least one active ingredient such as a muscadine pulp extract.

適用於所揭示之方法及組成物的醫藥學上可接受之載劑為此項技術中所習知。醫藥載劑之性質將視所用之特定投藥模式而定。舉例而言，非經腸調配物通常包含作為媒劑之可注射流體，包括醫藥學上及生理學上可接受之流體，諸如水、生理食鹽水、平衡鹽溶液、右旋糖水溶液、甘油或其類似物。對於諸如散劑、丸劑、錠劑或膠囊形式之固體組成物而言，習知無毒固體載劑可包括例如醫藥級甘露糖醇、乳糖、澱粉或硬脂酸鎂。除生物中性學載劑外，欲投予之醫藥組成物可視情況含有少量無毒輔助物質或賦形劑，諸如濕潤劑或乳化劑、防腐劑及pH值緩衝劑及其類似物；例如，乙酸鈉或脫水山梨糖醇單月桂酸酯。其他非限制性賦形劑包括非離子性增溶劑，諸如十六醇聚氧乙烯醚(cremophor)，或蛋白質，諸如人類血清白蛋白。Pharmaceutically acceptable carriers suitable for use in the disclosed methods and compositions are well known in the art. The nature of the pharmaceutical carrier will depend on the particular mode of administration employed. For example, parenteral formulations typically comprise an injectable fluid as a vehicle, including pharmaceutically and physiologically acceptable fluids such as water, physiological saline, balanced salt solutions, aqueous dextrose, glycerol or Its analogues. For solid compositions such as powders, pills, lozenges or capsules, conventional non-toxic solid carriers may include, for example, pharmaceutical grade mannitol, lactose, starch or magnesium stearate. In addition to the biological neutral carrier, the pharmaceutical composition to be administered may optionally contain minor amounts of non-toxic auxiliary substances or excipients such as wetting or emulsifying agents, preservatives and pH buffers and the like; for example, acetic acid Sodium or sorbitan monolaurate. Other non-limiting excipients include nonionic solubilizing agents such as cremote, or proteins such as human serum albumin.

所揭示之醫藥組成物可調配為醫藥學上可接受之鹽。醫藥學上可接受之鹽為具有游離鹼之所需藥理活性之游離鹼形式之化合物的無毒鹽。此等鹽可衍生自無機酸或有機酸。合適無機酸的非限制性實例為鹽酸、硝酸、氫溴酸、硫酸、氫碘酸及磷酸。合適有機酸的非限制性實例為乙酸、丙酸、乙醇酸、乳酸、丙酮酸、丙二酸、丁二酸、蘋果酸、順丁烯二酸、反丁烯二酸、酒石酸、檸檬酸、苯甲酸、肉桂酸、扁桃酸、甲烷磺酸、乙烷磺酸、對甲苯磺酸、甲磺酸、水楊酸、甲酸、三氯乙酸、三氟乙酸、葡萄糖酸、天冬胺酸(asparagic acid/aspartic acid)、苯磺酸、對甲苯磺酸、萘磺酸及其類似酸。其他合適醫藥學上可接受之鹽的清單見於Remington’s Pharmaceutical Sciences ,第19版,Mack Publishing公司,Easton,PA,1995中。醫藥學上可接受之鹽亦可用以調節組成物之滲透壓。The disclosed pharmaceutical compositions can be formulated as pharmaceutically acceptable salts. A pharmaceutically acceptable salt is a non-toxic salt of a compound in free base form having the desired pharmacological activity of the free base. These salts can be derived from inorganic or organic acids. Non-limiting examples of suitable inorganic acids are hydrochloric acid, nitric acid, hydrobromic acid, sulfuric acid, hydroiodic acid, and phosphoric acid. Non-limiting examples of suitable organic acids are acetic acid, propionic acid, glycolic acid, lactic acid, pyruvic acid, malonic acid, succinic acid, malic acid, maleic acid, fumaric acid, tartaric acid, citric acid, Benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, methanesulfonic acid, salicylic acid, formic acid, trichloroacetic acid, trifluoroacetic acid, gluconic acid, aspartic acid (asparagic) Acid/aspartic acid), benzenesulfonic acid, p-toluenesulfonic acid, naphthalenesulfonic acid and the like. A list of other suitable pharmaceutically acceptable salts is found in Remington's Pharmaceutical Sciences , 19th Edition, Mack Publishing Company, Easton, PA, 1995. Pharmaceutically acceptable salts can also be used to adjust the osmotic pressure of the composition.

所揭示之醫藥組成物的劑型將藉由所選投藥模式來確定。舉例而言，可使用口服劑型。口服調配物可為液體，諸如糖漿、溶液或懸浮液；或固體，諸如散劑、丸劑、錠劑或膠囊。製備該等劑型之方法在本文中揭示，或其對於熟習此項技術者將顯而易見。The dosage form of the disclosed pharmaceutical composition will be determined by the mode of administration selected. For example, an oral dosage form can be used. The oral formulation can be a liquid, such as a syrup, solution or suspension; or a solid, such as a powder, pill, lozenge or capsule. Methods of preparing such dosage forms are disclosed herein or as will be apparent to those skilled in the art.

可以適合於個別投予精確劑量之單位劑型調配包含所揭示之化合物的醫藥組成物之某些具體實例。所投予之活性成份的量將視所治療之個體、病症之嚴重程度及投藥方式而定且為熟習此項技術者所已知。在此等範圍內，欲投予之調配物將含有一定量的本文所揭示之萃取物或化合物，其量有效達成所治療之個體體內的所需作用。在特定實施例中，對於經口投藥而言，組成物以含有約75mg麝香葡萄果漿萃取物多酚及100mg反式白藜蘆醇之液體形式提供。在其他實施例中，對於經口投藥而言，組成物以含有約1.0mg至約500mg活性成份、尤其約10.0mg、約50mg、100mg、約150mg、約175mg、約200mg、約210mg、約225mg、約250mg、約300mg、約400mg或約500mg活性成份之錠劑形式提供以用於按症狀調整投予所治療之個體的劑量。在一例示性經口給藥方案中，每日投予含有約1mg至約500mg(諸如約150mg至約225mg)活性成份之錠劑至少一次。在其他實施例中，每週投予含有約1mg至約500mg(諸如約150mg至約225mg)活性成份之錠劑至少一次，諸如每週或每日至少兩次、三次、至少四次、至少五次或至少六次。Certain specific examples of pharmaceutical compositions comprising the disclosed compounds may be formulated in unit dosage forms suitable for the precise administration of the precise dosage. The amount of active ingredient administered will depend on the individual being treated, the severity of the condition, and the mode of administration, and are known to those skilled in the art. Within such ranges, the formulation to be administered will contain an amount of the extract or compound disclosed herein in an amount effective to achieve the desired effect in the subject being treated. In a particular embodiment, for oral administration, the composition is provided as a liquid containing about 75 mg of muscadine pulp extract polyphenol and 100 mg of trans-resveratrol. In other embodiments, for oral administration, the composition contains from about 1.0 mg to about 500 mg of active ingredient, especially about 10.0 mg, about 50 mg, 100 mg, about 150 mg, about 175 mg, about 200 mg, about 210 mg, about 225 mg. A dosage form of about 250 mg, about 300 mg, about 400 mg or about 500 mg of the active ingredient is provided for symptomatic adjustment of the dosage administered to the individual being treated. In one exemplary oral administration regimen, a lozenge containing from about 1 mg to about 500 mg (such as from about 150 mg to about 225 mg) of the active ingredient is administered at least once a day. In other embodiments, a lozenge containing from about 1 mg to about 500 mg (such as from about 150 mg to about 225 mg) of active ingredient is administered at least once a week, such as at least twice, three times, at least four times, at least five per week or daily. Times or at least six times.

G.　所揭示之萃取物及組成物的投藥G. Administration of extracts and compositions disclosed

投予任一所揭示之組成物及萃取物的方法為常規方法。舉例而言，所揭示之方法(諸如包括用於防止或減少一或多個與衰老相關之過程的具有抗氧化劑活性之組成物的方法)可經由注射、腫瘤內、經口、局部、經皮、非經腸或經由吸入或噴霧來投予。在一特定實施例中，向哺乳動物個體(諸如人類)經口投予呈非飲料食品、飲料或膳食補充劑形式之組成物。在另一實施例中，向哺乳動物個體(諸如人類)之皮膚表面局部投予組成物。The method of administering any of the disclosed compositions and extracts is a conventional method. For example, the disclosed methods, such as methods comprising a composition having antioxidant activity for preventing or reducing one or more processes associated with aging, can be via injection, intratumoral, oral, topical, transdermal , administered parenterally or via inhalation or spray. In a particular embodiment, a mammalian individual, such as a human, is orally administered as a composition in the form of a non-beverage food, beverage or dietary supplement. In another embodiment, the composition is topically administered to the surface of the skin of a mammalian subject, such as a human.

所投予之藥劑的治療有效量可視所需作用及欲治療之個體而變化。在一實施例中，該方法包括向個體(諸如人類個體)每日投予至少1mg組成物。舉例而言，可向人類每日投予至少1g或至少10g組成物，諸如每日1g至5g、每日5g至10g，例如每日7g。在一實施例中，向個體投予至少5g包括麝香葡萄果漿萃取物及白藜蘆醇之組成物。在其他實施例中，向個體經口投予至少6.3g該組成物。劑量可以分次給藥形式(諸如每天分2、3或4次給藥)或以每日單一劑量形式投予。The therapeutically effective amount of the agent administered can vary depending on the desired effect and the individual to be treated. In one embodiment, the method comprises administering to the individual (such as a human subject) at least 1 mg of the composition per day. For example, at least 1 g or at least 10 g of the composition can be administered daily to humans, such as 1 g to 5 g per day, 5 g to 10 g per day, for example 7 g per day. In one embodiment, at least 5 grams of the composition comprising the muscadine pulp extract and resveratrol are administered to the individual. In other embodiments, at least 6.3 g of the composition is administered orally to an individual. The dose may be administered in divided doses (such as 2, 3 or 4 doses per day) or in a single daily dose.

在特定實施例中，以多個每日給藥時程(諸如至少連續2天、連續10天等等)向個體投予治療性組成物(諸如所揭示之包括白藜蘆醇及麝香葡萄萃取物的抗衰老補充劑)，例如歷時數週、數月或數年之時間。在一實施例中，向個體投予治療性組成物(諸如所揭示之包括白藜蘆醇及麝香葡萄萃取物的抗衰老補充劑)，包括每日投予，歷時至少30天之時間，諸如至少2個月、至少4個月、至少6個月、至少12個月、至少24個月或至少36個月。In a particular embodiment, the therapeutic composition is administered to the individual in a plurality of daily dosing schedules (such as at least 2 consecutive days, for 10 consecutive days, etc.), such as the resveratrol and muscat grape extracts disclosed. Anti-aging supplements, for example, for weeks, months or years. In one embodiment, the subject is administered a therapeutic composition (such as the disclosed anti-aging supplement comprising resveratrol and muscadine extract), including daily administration for at least 30 days, such as At least 2 months, at least 4 months, at least 6 months, at least 12 months, at least 24 months, or at least 36 months.

諸如所揭示之包括反式白藜蘆醇及麝香葡萄果漿萃取物或單獨麝香葡萄果漿萃取物的抗衰老補充劑之治療性組成物可進一步包括一或多種生物活性或非活性化合物(或兩者)，分別諸如其他抗氧化劑及習知無毒醫藥學上可接受之載劑。在一特定實施例中，治療性組成物進一步包括一或多種生物非活性化合物。該等生物非活性化合物之實例包括(但不限於)：載劑、增稠劑、稀釋劑、緩衝劑、防腐劑及載劑。適用於此等調配物之醫藥學上可接受之載劑為習知的(參見Remington’s Pharmaceutical Sciences,E. W. Martin,Mack Publishing公司,Easton,PA,第19版(1995))。一般而言，載劑之性質將視所用特定投藥模式而定。舉例而言，非經腸調配物可包括作為媒劑之可注射流體，包括醫藥學上及生理學上可接受之流體，諸如水、生理食鹽水、平衡鹽溶液、右旋糖水溶液、甘油或其類似物。對於固體組成物(例如，散劑、丸劑、錠劑或膠囊形式)而言，習知無毒固體載劑可包括例如醫藥級甘露糖醇、乳糖、澱粉或硬脂酸鎂。除生物中性載劑外，欲投予之醫藥組成物亦可包括少量無毒輔助物質，諸如濕潤劑或乳化劑、防腐劑及pH值緩衝劑及其類似物，例如乙酸鈉或脫水山梨糖醇單月桂酸酯。A therapeutic composition such as the disclosed anti-aging supplement comprising trans-resveratrol and muscadine pulp extract or muscadine pulp extract alone may further comprise one or more biologically active or inactive compounds (or Both), such as other antioxidants and conventional non-toxic pharmaceutically acceptable carriers. In a particular embodiment, the therapeutic composition further comprises one or more biologically inactive compounds. Examples of such biologically inactive compounds include, but are not limited to, carriers, thickeners, diluents, buffers, preservatives, and carriers. Pharmaceutically acceptable carriers suitable for use in such formulations are conventional (see Remington's Pharmaceutical Sciences, E. W. Martin, Mack Publishing Company, Easton, PA, 19th Edition (1995)). In general, the nature of the carrier will depend on the particular mode of administration employed. For example, a parenteral formulation can include an injectable fluid as a vehicle, including pharmaceutically and physiologically acceptable fluids such as water, physiological saline, balanced salt solutions, aqueous dextrose, glycerol or Its analogues. For solid compositions (eg, powders, pills, troches, or capsules), conventional non-toxic solid carriers can include, for example, pharmaceutical grade mannitol, lactose, starch, or magnesium stearate. In addition to the biologically neutral carrier, the pharmaceutical composition to be administered may also include minor amounts of non-toxic auxiliary substances such as wetting or emulsifying agents, preservatives and pH buffering agents and the like, such as sodium acetate or sorbitan. Monolaurate.

儘管本揭示案已提供使用任一所揭示之組成物及萃取物來防止或抑制一或多個與細胞衰老相關之過程的詳細說明，但預期所揭示之組成物及萃取物不僅可用作防止或抑制一或多個與細胞衰老相關之過程的抗氧化劑，而且可治療與氧化應力相關之任何病症。舉例而言，預期本發明之萃取物及組成物可用於減少、預防或治療與諸如糖尿病、癌症、動脈粥樣硬化及其他心血管疾病之慢性發炎性疾病的發病機制以及與諸如阿茲海默氏症及帕金森氏病之退化性疾病相關的氧化應力。Although the present disclosure has provided a detailed description of the use of any of the disclosed compositions and extracts to prevent or inhibit one or more processes associated with cellular senescence, it is contemplated that the disclosed compositions and extracts can be used not only as a preventative Or an antioxidant that inhibits one or more processes associated with cellular aging, and can treat any condition associated with oxidative stress. For example, it is contemplated that the extracts and compositions of the present invention are useful for reducing, preventing or treating the pathogenesis of chronic inflammatory diseases such as diabetes, cancer, atherosclerosis, and other cardiovascular diseases, as well as with, for example, Alzheimer's Oxidative stress associated with degenerative diseases of Parkinson's disease.

藉由以下非限制性實施例進一步說明本揭示案之標的物。The subject matter of the present disclosure is further illustrated by the following non-limiting examples.

實施例Example

實施例1Example 1

土耳其鞣酸在麝香葡萄果漿萃取物中之溶解度的測定Determination of Solubility of Turkish Tannin in Musk Grape Fruit Extract

該實施例說明青銅色與紫色果漿萃取物之各種比率對土耳其鞣酸溶解度之影響。This example illustrates the effect of various ratios of bronze and purple pulp extracts on the solubility of citric acid in Turkey.

加熱單獨醱酵之青銅色及紫色果漿萃取物且接著以表1中所提供之比率以100毫升之總體積混合。The separately fermented bronze and purple pulp extracts were heated and then mixed in a total volume of 100 ml at the ratios provided in Table 1.

因為未對單獨青銅色及紫色果漿萃取物進行濃縮，所以所得萃取物混合物為1×濃度。1×濃度典型地含有約2%固體溶液(100公克溶液等於2公克乾萃取物)。將約20毫升之各比率轉移至各別玻璃試管中且用箔覆蓋。將樣品於85℃下加熱30分鐘，接著移離熱源且使其達到室溫。接著經由0.45μm PTFE過濾器w/GMP過濾樣品且經由HPLC/MS分析以測定其土耳其鞣酸含量。亦經由相同方法分析單獨青銅色及紫色麝香葡萄果漿萃取物以測定土耳其鞣酸在青銅色及紫色果漿萃取物兩者中之基線值且測定紫色果漿萃取物中花青素之含量。接著藉由逆相HPLC在254nm下進行UV-VIS偵測，結合離子阱質量偵測器使用萃取離子層析圖(extracted ion chromatogram，EIC)在[M-H]^- =301amu下偵測，來測定樣品中土耳其鞣酸之總範圍。Since the individual bronze and purple pulp extracts were not concentrated, the resulting extract mixture was 1X concentration. The 1X concentration typically contains about 2% solids solution (100 grams of solution equals 2 grams of dry extract). Each of the approximately 20 ml ratios was transferred to individual glass tubes and covered with foil. The sample was heated at 85 ° C for 30 minutes, then removed from the heat source and allowed to reach room temperature. The sample was then filtered through a 0.45 μm PTFE filter w/GMP and analyzed by HPLC/MS to determine its Turkish tannic acid content. The individual bronze and purple muscadine pulp extracts were also analyzed by the same method to determine the baseline value of turmeric tannin in both the bronze and purple pulp extracts and to determine the anthocyanin content of the purple pulp extract. Then, UV-VIS detection was carried out by reverse phase HPLC at 254 nm, and the ion trap mass detector was used to detect the sample by using an extracted ion chromatogram (EIC) at [MH] ^- = 301 amu. The total range of citric acid in Turkey.

為製備具有40%固體之各種比率之青銅色/紫色果漿萃取物，將1×單獨青銅色及紫色麝香葡萄果漿萃取物乾燥為粉末形式且用水復原以產生含有40%固體之液體。接著根據重量(wt)而非體積將此兩種單獨溶液以各種比率混合。表2A及2B中提供所評估之各種比率。To prepare a bronze/purple pulp extract having various ratios of 40% solids, 1 x individual bronze and purple muscadine pulp extracts were dried to a powder form and reconstituted with water to produce a liquid containing 40% solids. The two separate solutions are then mixed in various ratios depending on the weight (wt) rather than the volume. The various ratios evaluated are provided in Tables 2A and 2B.

將約10毫升之各比率轉移至各別玻璃試管中且用箔覆蓋。將樣品於85℃下加熱30分鐘，接著移離熱源且使其達到室溫。接著經由0.45μm PTFE過濾器w/GMP過濾樣品且經由HPLC/MS分析以測定其土耳其鞣酸含量。亦經由相同方法分析單獨青銅色及紫色麝香葡萄果漿萃取物以測定土耳其鞣酸在青銅色及紫色麝香葡萄果漿萃取物兩者中之基線值且測定紫色麝香葡萄果漿萃取物中花青素之含量。結合離子阱質量偵測器，使用萃取離子層析圖(EIC)在[M-H]^- =301amu下偵測，使用驗證測試方法C2505來測定樣品中土耳其鞣酸之總範圍。Each ratio of about 10 ml was transferred to a separate glass test tube and covered with a foil. The sample was heated at 85 ° C for 30 minutes, then removed from the heat source and allowed to reach room temperature. The sample was then filtered through a 0.45 μm PTFE filter w/GMP and analyzed by HPLC/MS to determine its Turkish tannic acid content. The individual bronze and purple musk grape pulp extracts were also analyzed by the same method to determine the baseline value of the Turkish tannic acid in both the bronze and purple musk grape pulp extracts and to determine the purple color in the purple musk grape pulp extract. The content of the prime. Combined with the ion trap mass detector, the extracted ion chromatogram (EIC) was used to detect at [MH] ^- = 301 amu, and the verification test method C2505 was used to determine the total range of citric acid in the sample.

此等研究發現65%/35%之青銅色與紫色果漿萃取物(體積比體積)之比率足以提高土耳其鞣酸在1×濃度值之兩種果漿萃取物之混合物中的最大溶解度。此外，於40%固體含量下，土耳其鞣酸之溶解度繼續隨混合物中紫色麝香葡萄果漿萃取物之含量增加而增加。此等發現表明於40%固體含量下，較大量之土耳其鞣酸以固體形式存在，因此需要較高含量之紫色果漿萃取物以使混合物中所存在之所有土耳其鞣酸完全溶解。此等研究亦可表明紫色麝香葡萄果漿萃取物中所含之花青素影響青銅色麝香葡萄果漿萃取物中所含之土耳其鞣酸的溶解度。These studies found that the ratio of 65%/35% bronze to purple pulp extract (volume by volume) is sufficient to increase the maximum solubility of the Turkish tannic acid in a mixture of the two pulp extracts at a concentration of 1x. In addition, at 40% solids, the solubility of citric acid in Turkey continued to increase with the increase in the content of the purple muscadine pulp extract in the mixture. These findings indicate that at 40% solids, a larger amount of turmeric acid is present in solid form, so a higher level of purple pulp extract is required to completely dissolve all of the turmeric acid present in the mixture. These studies may also indicate that the anthocyanins contained in the purple muscadine pulp extract affect the solubility of the Turkish tannic acid contained in the bronze musk grape pulp extract.

實施例2Example 2

抗衰老膳食補充劑及其製備Anti-aging dietary supplement and preparation thereof

該實施例提供可食用以防止或抑制一或多個與細胞衰老相關之過程的膳食補充劑。This embodiment provides a dietary supplement that is edible to prevent or inhibit one or more processes associated with cellular aging.

根據表3a或表3b中所提供之配方來製備所揭示之抗衰老膳食補充劑。The disclosed anti-aging dietary supplements are prepared according to the formulations provided in Table 3a or Table 3b.

膳食補充劑包括關於劑量之說明書。該等說明書指示可每日在餐前服用5毫升補充劑(約一湯匙)且其用作抗衰老補充劑以例如抑制一或多個與細胞衰老相關之過程。Dietary supplements include instructions for dosage. These instructions indicate that 5 ml of supplement (about one tablespoon) can be taken daily before a meal and used as an anti-aging supplement to, for example, inhibit one or more processes associated with cellular aging.

實施例3Example 3

麝香葡萄果漿萃取物與日本蓼根萃取物之混合物及抗衰老補充劑的抗氧化能力Antioxidant ability of muscadine grape fruit extract and Japanese root extract and anti-aging supplement

該實施例表明含有日本蓼萃取物及麝香葡萄果漿萃取物之混合物及補充劑具有改良的抗氧化能力，如藉由氧自由基吸收能力(ORAC)檢定所量測。This example demonstrates that the mixture and supplement containing the Japanese cockroach extract and the muscadine pulp extract has improved antioxidant capacity, as measured by the Oxygen Free Radical Absorption Capacity (ORAC) assay.

使用ORAC檢定評估兩種植物萃取物單獨或以混合物或補充劑形式組合之抗氧化能力。已使用此檢定量測多種食品及飲料之抗氧化能力且該檢定為USDA ORAC資料庫中所含之資料的基礎。可藉由此測試量測親水性抗氧化能力與親脂性抗氧化能力兩者。The ORAC assay was used to assess the antioxidant capacity of the two plant extracts, either alone or in combination as a mixture or supplement. This assay has been used to quantify the antioxidant capacity of a variety of foods and beverages and this check is based on the information contained in the USDA ORAC database. Both hydrophilic antioxidant capacity and lipophilic antioxidant capacity can be measured by this test.

分析以下樣品：(1)標準化為最小98%反式白藜蘆醇之乾燥/粉末狀日本蓼根萃取物(實際含量為100%反式白藜蘆醇)；(2)含有14.4%總多酚含量之乾燥/粉末狀麝香葡萄果漿萃取物(青銅色與紫色果漿之比率為2:1)；(3)上述乾燥/粉末狀日本蓼與乾燥/粉末狀麝香葡萄果漿萃取物以1:5.36(wt:wt)比率之混合物(總多酚含量為混合物之27.5%且麝香葡萄多酚與反式白藜蘆醇之比率為0.75/1)；及(4)實施例2表3a中所提供之抗衰老補充劑。量測親水性、親脂性及總ORAC值(總ORAC值為親水性值與親脂性值之總和)且結果以每毫克多酚之微莫耳Trolox當量數(微莫耳TE/毫克多酚)表示。該等結果展示於表4中。The following samples were analyzed: (1) normalized to a minimum of 98% trans-resveratrol dry/powdered Japanese root extract (actual content is 100% trans-resveratrol); (2) containing 14.4% total Phenol content dry/powdered muscadine pulp extract (2:1 ratio of bronze to purple pulp); (3) dried/powdered Japanese glutinous rice and dried/powdered muscadine pulp extract a mixture of 1:5.36 (wt:wt) ratio (total polyphenol content is 27.5% of the mixture and the ratio of muscadine polyphenol to trans-resveratrol is 0.75/1); and (4) Table 2a of Example 2 An anti-aging supplement provided in the product. The hydrophilicity, lipophilicity and total ORAC value were measured (the total ORAC value is the sum of the hydrophilic value and the lipophilic value) and the result is the number of micromolar Trolox equivalents per milligram of polyphenol (micromolar TE/mg polyphenol) Said. These results are shown in Table 4.

如表4中所示，混合物的實測親水性ORAC值類似於基於兩種萃取物之相加效應的預測值。然而，混合物的實測親脂性ORAC值為預測相加值之5倍，導致總ORAC值增加20%。對於組成物觀察到類似協同作用，其接近基於相加性之預測ORAC計分的3倍。表5展示以每公克物質(與表4中所示之每毫克多酚相對比)的ORAC值表示之結果。如表5中所示，當以每公克物質的ORAC表示值時，麝香葡萄果漿萃取物與日本蓼根萃取物之混合物及抗衰老補充劑在產生親脂性抗氧化能力方面之協同效應被保持：As shown in Table 4, the measured hydrophilic ORAC value of the mixture was similar to the predicted value based on the additive effect of the two extracts. However, the measured lipophilic ORAC value of the mixture was 5 times the predicted additive value, resulting in a 20% increase in the total ORAC value. A similar synergistic effect was observed for the composition, which is close to three times the predicted ORAC score based on additive. Table 5 shows the results expressed as ORAC values per gram of material (as opposed to each milligram of polyphenol shown in Table 4). As shown in Table 5, when the value is expressed in ORAC per gram of the substance, the synergistic effect of the mixture of the muscadine pulp extract and the Japanese jaundice extract and the anti-aging supplement in the production of lipophilic antioxidant capacity is maintained. :

此等研究表明麝香葡萄果漿萃取物與日本蓼根萃取物之混合物及抗衰老補充劑在產生親脂性抗氧化能力方面之強協同效應。未預料到在親脂性條件下展現選擇性協同作用。因此，本發明之發現結果表明與任一單獨萃取物相比，如藉由ORAC檢定所量測，含有日本蓼萃取物及麝香葡萄果漿萃取物之混合物具有改良的抗氧化能力。These studies have shown a strong synergistic effect of the mixture of muscadine grape fruit extract and Japanese root extract and anti-aging supplements in producing lipophilic antioxidant capacity. It was not expected to exhibit selective synergy under lipophilic conditions. Thus, the findings of the present invention indicate that the mixture containing the Japanese cockroach extract and the muscadine pulp extract has improved antioxidant capacity as measured by the ORAC assay as compared to any of the individual extracts.

實施例4Example 4

抗氧化劑膠囊調配物及製備Antioxidant capsule formulation and preparation

該實施例提供抗氧化劑膠囊調配物及其製備，該調配物可用於抑制一或多個氧化過程，包括防止或抑制自由基形成。This embodiment provides an antioxidant capsule formulation and preparation thereof, which formulation can be used to inhibit one or more oxidation processes, including preventing or inhibiting free radical formation.

如下處理具有40%之固體含量及以固體計10%之多酚含量(以五倍子酸當量(Gallic Acid equivalent，GAE)量測)的比率為2:1之青銅色與紫色果漿之水性萃取物：將1.33Kg麥芽糊精MD100添加至10Kg上文所提及之果漿濃縮物中。摻合該混合物以使麥芽糊精溶解。隨後，冷凍乾燥此液體濃縮物以產生乾物質。如下為乾燥麝香葡萄/麥芽糊精物質之化學值：(1)總多酚(以%GAE計)，NLT 7.0%(乾基重)；五倍子酸(HPLC)，NLT 0.3%(乾基重)；及游離土耳其鞣酸(HPLC)，NLT 0.15%(乾基重)。表6中提供乾燥麝香葡萄/麥芽糊精之特徵。An aqueous extract of bronze and purple pulp with a ratio of solids content of 40% and a polyphenol content of 10% by solids (measured by Gallic Acid equivalent (GAE)) of 2:1 was treated as follows : 1.33 Kg of maltodextrin MD100 was added to 10 Kg of the pulp concentrate mentioned above. The mixture is blended to dissolve the maltodextrin. This liquid concentrate is then lyophilized to produce a dry matter. The chemical values of the dried muscadine/maltodextrin material are as follows: (1) total polyphenol (in % GAE), NLT 7.0% (dry basis weight); gallic acid (HPLC), NLT 0.3% (dry basis weight) And; free Turkish citric acid (HPLC), NLT 0.15% (dry basis weight). The characteristics of dried muscadine/maltodextrin are provided in Table 6.

藉由稱量出分批量之個別原料來製備膠囊。使該等原料穿過Comil研磨機，最後將麥芽糊精傳送穿過該研磨機。摻合經研磨之物質直至均勻且接著填充至囊封器上之「0」號兩片式膠囊中。表7中提供所製造之劑型的一實施例且表8中提供麝香葡萄/白藜蘆醇膠囊分析結果。Capsules are prepared by weighing out individual ingredients in divided batches. Passing these materials through Comil The grinder finally passes the maltodextrin through the grinder. The ground material was blended until uniform and then filled into a two-piece capsule of size "0" on the encapsulator. An example of the dosage form produced is provided in Table 7 and the results of the analysis of the muscadine/resveratrol capsules are provided in Table 8.

實施例5Example 5

人類個體體內之氧化應力的降低Reduction of oxidative stress in human individuals

該實施例表明如安慰劑對照之臨床試驗中所評估，實施例4中所述之抗氧化劑膠囊的活體內功效。This example demonstrates the in vivo efficacy of the antioxidant capsules described in Example 4 as assessed in a placebo-controlled clinical trial.

該等膠囊含有麝香葡萄果漿萃取物與日本蓼萃取物之混合物以提供每3個膠囊劑量75mg麝香葡萄多酚及100mg反式白藜蘆醇(表7及8)。已知膳食消耗短期增加氧化應力且產生發炎性反應。舉例而言，已顯示食用900kcal速食餐引起健康正常體重人類體內之循環單核細胞(mononuclear cell，MNC)及多型核白血球(polymorphonuclear leukocyte，PMN)中的活性氧物質(ROS)產生顯著增加(Aljada等人Am. J. Clin. Nutr. 79:682-690,2004)。此等變化伴隨著為NADPH氧化酶(ROS產生之介體)之次單元的p47phox蛋白質之MNC的表現升高。The capsules contained a mixture of muscadine pulp extract and Japanese cockroach extract to provide 75 mg of muscadine polyphenol and 100 mg of trans-resveratrol per 3 capsules (Tables 7 and 8). Dietary consumption is known to increase oxidative stress in the short term and produce an inflammatory response. For example, consumption of 900kcal fast-food meals has been shown to cause a significant increase in reactive oxygen species (ROS) in circulating mononuclear cells (MNCs) and polymorphonuclear leukocytes (PMNs) in healthy normal-weight humans. (Aljada et al . Am. J. Clin. Nutr. 79:682-690, 2004). These changes are accompanied by an increase in the expression of MNC of the p47phox protein, which is a subunit of NADPH oxidase (mediator of ROS production).

為測試在此餐後氧化應力之模型中抗氧化劑膠囊之作用，提供一組10位健康個體(年齡：37±4歲，BMI 22.6±0.5kg/m2)，在單獨兩天內，在餐前10分鐘食用910kcal速食餐及單一劑量(3個膠囊)之抗氧化劑調配物或安慰劑(3個膠囊)。在基線及在膳食攝取後1、3及5小時收集血樣。在各處理後，葡萄糖、胰島素及脂質之循環濃度類似，表明所揭示之抗氧化劑膠囊並不改變膳食之消化/吸收。然而，在膳食消耗後1小時，所揭示之抗氧化劑膠囊處理大大減弱MNC(15%對比安慰劑情況下之62%)與PMN(8%對比安慰劑情況下之64%)中ROS產生的增加。另外，膳食加安慰劑攝取引起MNC中p47phox之蛋白質含量相對於基線顯著增加148%(P<0.05)，而膳食加抗氧化劑膠囊攝取完全防止MNC p47phox含量之任何顯著變化(P<0.05，利用雙因子RMANOVA)。此外，與抗氧化應力轉錄因子Nrf-2之DNA結合活性實際上下降的安慰劑處理組相比，在膳食加抗氧化劑膠囊攝取後3小時，抗氧化應力轉錄因子Nrf-2之DNA結合活性相對於基線顯著增加150%(P<0.05，根據RMANOVA及雙因子RMANOVA)。Nrf-2與編碼內源性抗氧化酶(例如，麩胱甘肽-S-轉移酶)之多個基因的抗氧化劑反應元件(antioxidant response element，ARE)啟動子序列結合，且因此，在攝取所揭示之抗氧化劑調配物後Nrf-2之DNA結合活性增加表明上調多種細胞防禦之可能性。To test the role of antioxidant capsules in this model of post-prandial oxidative stress, a group of 10 healthy individuals (age: 37 ± 4 years, BMI 22.6 ± 0.5 kg/m2) were provided, in separate two days, before meals. Eat 910kcal fast food meal and single dose (3 capsules) of antioxidant formulation or placebo (3 capsules) for 10 minutes. Blood samples were collected at baseline and at 1, 3, and 5 hours after dietary intake. The circulating concentrations of glucose, insulin and lipid were similar after each treatment, indicating that the disclosed antioxidant capsules did not alter the digestion/absorption of the diet. However, one hour after dietary consumption, the disclosed antioxidant capsule treatment significantly attenuated the increase in ROS production in MNC (15% vs. 62% in placebo) and PMN (8% vs. 64% in placebo). . In addition, dietary plus placebo intake resulted in a significant 148% increase in protein content of p47phox in MNC relative to baseline (P < 0.05), whereas dietary plus antioxidant capsule intake completely prevented any significant change in MNC p47phox content (P < 0.05, using double Factor RMANOVA). In addition, compared with the placebo-treated group in which the DNA-binding activity of the antioxidant stress transcription factor Nrf-2 was actually decreased, the DNA-binding activity of the antioxidant stress transcription factor Nrf-2 was relatively 3 hours after the intake of the dietary antioxidant capsule. Significantly increased by 150% at baseline (P < 0.05, according to RMANOVA and two-factor RMANOVA). Nrf-2 binds to an antioxidant response element (ARE) promoter sequence encoding multiple genes of an endogenous antioxidant enzyme (eg, glutathione-S-transferase) and, therefore, is ingested An increase in the DNA binding activity of Nrf-2 following the disclosed antioxidant formulation indicates the possibility of upregulating multiple cellular defenses.

最後，在膳食及安慰劑攝取後，在3小時時，血漿內毒素(endotoxin)含量相對於基線顯著升高60%(P<0.05)，而當同時食用膳食與所揭示之抗氧化劑膠囊時，在1小時時，內毒素濃度相對於基線顯著下降28%(P<0.05，使用RMANOVA及雙因子RMANOVA)。內毒素為促使多種增強氧化應力之細胞激素活化的有效發炎信號。Finally, at 3 hours, plasma endotoxin levels were significantly increased by 60% (P < 0.05) relative to baseline after dietary and placebo ingestion, while when the diet was combined with the disclosed antioxidant capsules, At 1 hour, the endotoxin concentration decreased significantly by 28% relative to baseline (P < 0.05, using RMANOVA and two-way RMANOVA). Endotoxin is an effective inflammatory signal that promotes the activation of a variety of cytokines that enhance oxidative stress.

此等結果表明麝香葡萄果漿萃取物與日本蓼之混合物(麝香葡萄多酚與反式白藜蘆醇之比率為0.75/1)的新穎活體內抗氧化劑作用且因此延伸出此混合物之協同抗氧化能力的試管內發現(實施例3)。These results indicate a novel in vivo antioxidant effect of the mixture of muscadine grape puree extract and Japanese glutinous rice (the ratio of muscadine polyphenols to trans-resveratrol is 0.75/1) and thus extend the synergistic resistance of this mixture. The oxidizing ability was found in a test tube (Example 3).

鑒於可應用本揭示案之原理的許多可能具體實例，應認識到所說明之實施例僅為所揭示內容之實施例且不應視為限制本揭示案之範疇。更確切而言，本揭示案之範疇由以下申請專利範圍界定。因此，吾等主張吾等之發明全部均在此等申請專利範圍之範疇及精神內。In view of the many possible specific examples to which the principles of the present disclosure may be applied, it is to be understood that the illustrated embodiments are only examples of the disclosed embodiments and should not be construed as limiting the scope of the disclosure. Rather, the scope of the disclosure is defined by the scope of the following claims. Accordingly, we have claimed that all of our inventions are within the scope and spirit of such patent application.

Claims (32)

一種製備抗氧化劑口服膳食補充劑組成物之方法，其包含：將具有至少2%之多酚含量的麝香葡萄(圓葉葡萄(Vitis rotundifolia ))果漿液體溶劑萃取物與來自麝香葡萄以外的來源之具有最小純度為至少5%反式白藜蘆醇(trans-resveratrol)的白藜蘆醇(resveratrol)組合，其中麝香葡萄果漿萃取物多酚與反式白藜蘆醇之比率在0.1/1至10/1(重量/重量)之範圍內，由此產生為口服膳食補充劑組成物之具有抗氧化劑活性之麝香葡萄果漿萃取物與反式白藜蘆醇之混合物。A method of preparing an antioxidant oral dietary supplement composition comprising: a liquid solvent extract of muscadine ( Vitis rotundifolia ) pulp having at least 2% polyphenol content and a source other than muscat a resveratrol combination having a minimum purity of at least 5% trans-resveratrol, wherein the ratio of the polyphenol to trans-resveratrol in the muscadine pulp extract is 0.1/ From 1 to 10/1 (weight/weight), a mixture of muscadine pulp extract and trans-resveratrol having antioxidant activity as an oral dietary supplement composition is thereby produced. 如申請專利範圍第1項之方法，其中白藜蘆醇包含自日本蓼(虎杖(Polygonum cuspidatum )根萃取之至少98%反式白藜蘆醇。The method of claim 1, wherein the resveratrol comprises at least 98% trans-resveratrol extracted from the roots of Polygonum cuspidatum . 如申請專利範圍第1或2項之方法，其中該麝香葡萄(圓葉葡萄)果漿萃取物具有約4%之總多酚含量且該麝香葡萄果漿萃取物與該白藜蘆醇之比率為約18/1(重量/重量)。 The method of claim 1 or 2, wherein the muscadine (bougainvillea) pulp extract has a total polyphenol content of about 4% and the ratio of the muscadine pulp extract to the resveratrol It is about 18/1 (weight/weight). 如申請專利範圍第1項之方法，其進一步包含製備該麝香葡萄果漿萃取物，隨後將該麝香葡萄(圓葉葡萄)果漿萃取物與該白藜蘆醇組合。 The method of claim 1, further comprising preparing the muscadine pulp extract, and subsequently combining the muscadine (musk) pulp extract with the resveratrol. 如申請專利範圍第4項之方法，其中製備該麝香葡萄果漿萃取物包含將青銅色麝香葡萄果漿萃取物與紫色麝香葡萄果漿萃取物組合，且其中青銅色麝香葡萄果漿萃取物 與紫色麝香葡萄果漿萃取物之混合物促進麝香葡萄果漿萃取物中土耳其鞣酸(ellagic acid)的溶解度。 The method of claim 4, wherein the extracting the muscadine pulp extract comprises combining a bronze muscadine pulp extract with a purple muscadine pulp extract, and wherein the bronze muscadine pulp extract is The mixture with the purple muscadine pulp extract promotes the solubility of turkey ellagic acid in the muscadine pulp extract. 如申請專利範圍第4項之方法，其中製備該麝香葡萄果漿萃取物包含同時萃取紫色麝香葡萄果漿與青銅色麝香葡萄果漿之混合物以促進麝香葡萄果漿萃取物中土耳其鞣酸的溶解度。 The method of claim 4, wherein the extracting the muscadine pulp extract comprises simultaneously extracting a mixture of purple muscadine pulp and bronze muscadine pulp to promote the solubility of citric acid in the muscadine pulp extract. . 如申請專利範圍第6項之方法，其中青銅色麝香葡萄果漿萃取物與紫色麝香葡萄果漿萃取物之比率在0.3至3(重量/重量)之範圍內。 The method of claim 6, wherein the ratio of the bronze muscadine pulp extract to the purple muscadine pulp extract is in the range of 0.3 to 3 (weight/weight). 如申請專利範圍第6或7項之方法，其中該麝香葡萄果漿萃取物包含於液體中之20%至50%固體。 The method of claim 6 or 7, wherein the muscadine pulp extract comprises 20% to 50% solids in the liquid. 如申請專利範圍第8項之方法，其中該麝香葡萄果漿萃取物包含於液體中之約40%固體。 The method of claim 8, wherein the muscadine pulp extract comprises about 40% solids in the liquid. 如申請專利範圍第1或2項之方法，其進一步包含將接骨木萃取物、紫胡蘿蔔萃取物或其組合添加至該麝香葡萄果漿萃取物與白藜蘆醇之混合物中。 The method of claim 1 or 2, further comprising adding the elderberry extract, the purple carrot extract or a combination thereof to the mixture of the muscadine pulp extract and resveratrol. 如申請專利範圍第1或2項之方法，其進一步包含將賦形劑添加至該麝香葡萄果漿萃取物與白藜蘆醇之混合物中。 The method of claim 1 or 2, further comprising adding an excipient to the mixture of the muscadine pulp extract and resveratrol. 如申請專利範圍第11項之方法，其中該賦形劑包含甘油、山梨糖醇、膠態二氧化矽、天然調味添加劑或其組合。 The method of claim 11, wherein the excipient comprises glycerin, sorbitol, colloidal cerium oxide, a natural flavoring additive, or a combination thereof. 如申請專利範圍第5或6項之方法，其中該青銅色麝香葡萄果漿萃取物與紫色麝香葡萄果漿萃取物為經過濾 且經發酵之萃取物。 The method of claim 5, wherein the bronze muscadine pulp extract and the purple musk grape pulp extract are filtered. And the fermented extract. 一種用作抗氧化劑之口服膳食補充劑組成物，其包含具有至少2%之多酚含量的麝香葡萄(圓葉葡萄)果漿溶劑萃取物及來自麝香葡萄以外的來源之具有最小純度為至少5%反式白藜蘆醇的白藜蘆醇，其中麝香葡萄果漿萃取物多酚與反式白藜蘆醇之比率在0.1/1至10/1(重量/重量)之範圍內，由此提供具有抗氧化劑活性之口服膳食補充劑組成物。 An oral dietary supplement composition for use as an antioxidant comprising a muscadine (circle grape) pulp extract having at least 2% polyphenol content and a source other than muscadine having a minimum purity of at least 5 Resveratrol of trans-resveratrol, wherein the ratio of polyphenols to trans-resveratrol in the muscadine pulp extract is in the range of 0.1/1 to 10/1 (weight/weight), thereby An oral dietary supplement composition having antioxidant activity is provided. 如申請專利範圍第14項之口服膳食補充劑組成物，其中白藜蘆醇包含自日本蓼(虎杖(Polygonum cuspidatum )根萃取之至少98%反式白藜蘆醇。An oral dietary supplement composition according to claim 14, wherein the resveratrol comprises at least 98% trans-resveratrol extracted from the roots of Polygonum cuspidatum . 如申請專利範圍第14或15項之口服膳食補充劑組成物，其中該麝香葡萄(圓葉葡萄)果漿萃取物具有約4%之總多酚含量且該麝香葡萄果漿萃取物與該白藜蘆醇之比率為約18/1(重量/重量)。 An oral dietary supplement composition according to claim 14 or 15, wherein the muscadine (round leaf grape) pulp extract has a total polyphenol content of about 4% and the muscadine pulp extract and the white The ratio of resveratrol is about 18/1 (weight/weight). 如申請專利範圍第14或15項之口服膳食補充劑組成物，其中該麝香葡萄果漿萃取物包含自青銅色麝香葡萄果漿及紫色麝香葡萄果漿獲得之麝香葡萄果漿萃取物之組合以促進麝香葡萄果漿萃取物中土耳其鞣酸的溶解度。 The oral dietary supplement composition of claim 14 or 15, wherein the muscadine pulp extract comprises a combination of muscadine pulp extract obtained from bronze muscadine pulp and purple muscadine pulp. Promotes the solubility of citric acid in muscadine grape fruit extract. 如申請專利範圍第17項之口服膳食補充劑組成物，其中青銅色麝香葡萄果漿萃取物與紫色麝香葡萄果漿萃取物之比率在0.3至3(重量/重量)之範圍內。 The oral dietary supplement composition of claim 17, wherein the ratio of the bronze muscadine pulp extract to the purple muscadine pulp extract is in the range of 0.3 to 3 (weight/weight). 如申請專利範圍第17項之口服膳食補充劑組成物，其中該麝香葡萄果漿萃取物包含於液體中之20%至50% 固體。 The oral dietary supplement composition of claim 17, wherein the muscadine pulp extract is contained in the liquid in an amount of 20% to 50% solid. 如申請專利範圍第19項之口服膳食補充劑組成物，其中該麝香葡萄果漿萃取物包含於液體中之約40%固體。 The oral dietary supplement composition of claim 19, wherein the muscadine pulp extract comprises about 40% solids in the liquid. 如申請專利範圍第14或15項之口服膳食補充劑組成物，其進一步包含接骨木萃取物、紫胡蘿蔔萃取物或其組合。 The oral dietary supplement composition of claim 14 or 15, further comprising an elderberry extract, a purple carrot extract or a combination thereof. 如申請專利範圍第14或15項之口服膳食補充劑組成物，其進一步包含賦形劑。 The oral dietary supplement composition of claim 14 or 15 further comprising an excipient. 如申請專利範圍第22項之口服膳食補充劑組成物，其中該賦形劑包含甘油、山梨糖醇、膠態二氧化矽、天然調味添加劑或其組合。 The oral dietary supplement composition of claim 22, wherein the excipient comprises glycerin, sorbitol, colloidal cerium oxide, a natural flavoring additive, or a combination thereof. 如申請專利範圍第14或15項之口服膳食補充劑組成物，其中該組成物具有每毫克多酚至少24微莫耳Trolox當量(微莫耳TE/毫克多酚)之總氧自由基吸收能力(Oxygen Radical Absorbance Capacity，ORAC)。 An oral dietary supplement composition according to claim 14 or 15, wherein the composition has a total oxygen radical absorption capacity of at least 24 micromoles of Trolox equivalent per microgram of polyphenol (micromolar TE/mg polyphenol) (Oxygen Radical Absorbance Capacity, ORAC). 如申請專利範圍第14或15項之口服膳食補充劑組成物，其中該組成物提供於非飲料食品、飲料、液體或固體膳食補充劑中。 The oral dietary supplement composition of claim 14 or 15, wherein the composition is provided in a non-beverage food, beverage, liquid or solid dietary supplement. 如申請專利範圍第14或15項之口服膳食補充劑組成物，其係用作抗衰老補充劑。 An oral dietary supplement composition as claimed in claim 14 or 15 which is used as an anti-aging supplement. 如申請專利範圍第14或15項之口服膳食補充劑組成物，其中該組成物具有約0.75/1(重量/重量)之麝香葡萄多酚與白藜蘆醇的比率。 The oral dietary supplement composition of claim 14 or 15, wherein the composition has a ratio of muscadine polyphenol to resveratrol of about 0.75/1 (weight/weight). 如申請專利範圍第17項之口服膳食補充劑組成物，其中該青銅色麝香葡萄果漿萃取物與紫色麝香葡萄果漿萃取物為經過濾且經發酵之萃取物。 The oral dietary supplement composition of claim 17, wherein the bronze muscadine pulp extract and the purple musk grape pulp extract are filtered and fermented extracts. 一種用於抑制細胞衰老之組成物，其包含足以抑制一或多個與細胞衰老相關之過程之劑量的如申請專利範圍第14至28項中任一項之口服膳食補充劑組成物。 A composition for inhibiting cell senescence comprising an oral dietary supplement composition according to any one of claims 14 to 28, which comprises a dose sufficient to inhibit one or more processes associated with cell senescence. 如申請專利範圍第29項之組成物，其中抑制一或多個與細胞衰老相關之過程包含防止或抑制攝取該組成物之個體體內的自由基形成或活性。 The composition of claim 29, wherein the inhibiting one or more of the processes associated with cellular senescence comprises preventing or inhibiting free radical formation or activity in an individual ingesting the composition. 一種如申請專利範圍第14至28項中任一項之口服膳食補充劑組成物的用途，其係用於製造抑制細胞衰老的醫藥品，其中該醫藥品中的組成物之劑量是足以抑制一或多個與細胞衰老相關之過程。 The use of an oral dietary supplement composition according to any one of claims 14 to 28, which is for the manufacture of a medicament for inhibiting cell senescence, wherein the dose of the composition in the medicament is sufficient to inhibit one Or multiple processes associated with cellular aging. 如申請專利範圍第31項之用途，其中抑制一或多個與細胞衰老相關之過程包含防止或抑制攝取該組成物之個體體內的自由基形成或活性。 The use of claim 31, wherein inhibiting one or more of the processes associated with cellular senescence comprises preventing or inhibiting free radical formation or activity in an individual ingesting the composition.