WO2020118576A1 - 胚芽乳酸杆菌twk10在制备抗运动后发炎或降体脂组合物的用途 - Google Patents

胚芽乳酸杆菌twk10在制备抗运动后发炎或降体脂组合物的用途 Download PDF

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WO2020118576A1
WO2020118576A1 PCT/CN2018/120722 CN2018120722W WO2020118576A1 WO 2020118576 A1 WO2020118576 A1 WO 2020118576A1 CN 2018120722 W CN2018120722 W CN 2018120722W WO 2020118576 A1 WO2020118576 A1 WO 2020118576A1
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twk10
lactobacillus
probiotic composition
use according
body fat
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PCT/CN2018/120722
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French (fr)
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黄启彰
黄文经
林金生
李孟谦
黄可沁
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生合生物科技股份有限公司
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Priority to EP18942982.2A priority Critical patent/EP3896151A4/en
Priority to JP2021526475A priority patent/JP7130287B2/ja
Priority to KR1020217014456A priority patent/KR102606952B1/ko
Priority to PCT/CN2018/120722 priority patent/WO2020118576A1/zh
Publication of WO2020118576A1 publication Critical patent/WO2020118576A1/zh

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    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
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    • A61K35/66—Microorganisms or materials therefrom
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    • A61K35/741—Probiotics
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    • A61K35/74—Bacteria
    • A61K35/741—Probiotics
    • A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
    • A61K35/747—Lactobacilli, e.g. L. acidophilus or L. brevis
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    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
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    • A—HUMAN NECESSITIES
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    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/04—Anorexiants; Antiobesity agents
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    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
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    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • C12N1/205—Bacterial isolates
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2200/00—Function of food ingredients
    • A23V2200/30—Foods, ingredients or supplements having a functional effect on health
    • A23V2200/324—Foods, ingredients or supplements having a functional effect on health having an effect on the immune system
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2200/00—Function of food ingredients
    • A23V2200/30—Foods, ingredients or supplements having a functional effect on health
    • A23V2200/332—Promoters of weight control and weight loss
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/11—Lactobacillus
    • A23V2400/169—Plantarum
    • A—HUMAN NECESSITIES
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    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K2035/11—Medicinal preparations comprising living procariotic cells
    • A61K2035/115—Probiotics
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    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C12R2001/225—Lactobacillus
    • C12R2001/25—Lactobacillus plantarum

Definitions

  • the present invention is an application field of probiotics, in particular to describe that the Lactobacillus germ can be used to resist inflammation or reduce body fat after exercise.
  • Probiotics are natural sources, no harm to the human body, low intake threshold, and have a variety of body health, regulating effects, so it is very popular in the market of healthy food.
  • Lactobacillus plantarum is a common probiotic. It is a gram-positive bacterium that can be found in fermented food or animal saliva. It has the largest gene body in lactic acid bacteria and has many variations, so it is often due to the degree of variation. Different from adaptation conditions, the same strains will show different characteristics and efficacy.
  • Lactobacillus germ has multiple medical uses, including viral infections to protect the respiratory tract, and mixing with chemotherapy drugs can increase the sensitivity of cancer cells to drugs, and there are many uses of food additives, such as to prevent growth A poisonous fungus used in corn.
  • Inflammation refers to the defensive response of living tissue with vascular system to inflammatory factors and local injury, which can be divided into acute inflammation and chronic inflammation.
  • Acute inflammation is the initial reaction of an organism in the face of harmful stimuli. More plasma and white blood cells (especially granulocytes) move from the blood to the damaged tissue to carry out a series of chain reactions that involve the local vascular system and immune system. And cells in the damaged tissue.
  • Chronic inflammation leads to changes in cell types at the site of inflammation, which simultaneously destroys and repairs the tissue.
  • human fat has the ability to secrete inflammatory hormones, so excessive body fat is also a factor that causes chronic inflammation in the body.
  • inflammation can even cause allergies in the body's own immune system, which in turn attacks its own tissues and cells, such as rheumatoid arthritis and lupus erythematosus.
  • Long-term chronic inflammation can cause a series of diseases, which will gradually make organs and tissues lose function, not only easily cause fatigue, aging, hormonal imbalance, but also cause various serious diseases, including hepatitis, pneumonia, nephritis, autoimmune diseases, arthritis , Diabetes, Alzheimer's disease, cardiovascular disease, and even cancer.
  • the present invention provides the use of a Lactobacillus germ TWK10 for the preparation of a probiotic composition for improving inflammation after exercise.
  • the Lactobacillus germ TWK10 is deposited at the Taiwan Food Industry Development Institute under the deposit number BCRC910734, and is deposited in the Chinese general microorganism Deposit Management Center (CGMCC), the deposit number is CGMCC 13008.
  • CGMCC Chinese general microorganism Deposit Management Center
  • the Lactobacillus germ TWK10 improves inflammation after exercise by reducing the ratio of neutrophil/lymphocyte ratio (NLR) and platelet/lymphocyte (PLR).
  • NLR neutrophil/lymphocyte ratio
  • PLR platelet/lymphocyte
  • the Lactobacillus germ TWK10 is a live or non-activated bacterium, and the non-activated bacterium is heat-killed or other physical or chemical treatments.
  • the dose is greater than or equal to 3x10 10 CFU/day; wherein, when the dose is greater than 1x10 11 CFU/day, it has a better effect of improving inflammation after exercise.
  • the present invention also provides a method for improving inflammation after exercise by taking an effective amount of a probiotic composition comprising the Lactobacillus germ TWK10 with the deposit number CGMCC 13008.
  • the present invention further provides a method for improving diseases caused by excessive neutrophil/lymphocyte ratio and platelet/lymphocyte ratio, which is to take an effective amount of a probiotic composition, which contains the deposit number Lactobacillus germ TWK10 of CGMCC 13008.
  • the present invention provides the use of a Lactobacillus germ TWK10 for the preparation of a probiotic composition for lowering body fat.
  • the Lactobacillus germ TWK10 is deposited at the Taiwan Food Industry Development Institute under the deposit number BCRC910734, and is deposited in the Chinese common microbial strains Deposit Management Center (CGMCC), deposit number is CGMCC 13008.
  • CGMCC Chinese common microbial strains Deposit Management Center
  • the Lactobacillus germ TWK10 lowers body fat to reduce whole body fat.
  • the probiotic bacteria included in the probiotic composition are live bacteria, non-activated bacteria, or a combination of the two; the dosage of the Lactobacillus germ TWK10 is greater than or equal to 3x10 10 CFU/day.
  • the present invention further provides a method of lowering body fat by taking an effective amount of a probiotic composition, the probiotic composition comprising Lactobacillus germ TWK10 with the deposit number CGMCC 13008.
  • the present invention further provides a method for improving diseases caused by excessive body fat by taking an effective amount of a probiotic composition comprising the Lactobacillus germ TWK10 with the deposit number CGMCC 13008.
  • the aforementioned probiotic compositions can all include pharmaceutically acceptable carriers, excipients, diluents, and the dosage form is selected from solutions, suspensions, emulsions, powders, lozenges, pills, syrups, buccal tablets, tablets , Chewing gum, thick pulp, and capsules.
  • the aforementioned probiotic composition can be further made into fluid dairy products, concentrated milk, yogurt, yogurt, frozen yogurt, lactobacillus fermented beverages, milk powder, ice cream, cheese, cheese, soy milk, fermented soy milk, vegetable juices, fruit juices, sports Beverage, dessert, jelly, candy, baby food, health food, animal feed, Chinese herbal medicine composition, dietary supplement.
  • the aforementioned probiotic composition includes one or any combination of the following probiotic groups: Lactobacillus species, Lactococcus species, Pediococcus species, Leuconostoc species, Enterococcus species, Streptococcus species , Bifidobacterium species, yeast.
  • An example of the present invention shows that both the TWK10 live bacteria high-dose group (3x10 11 CFU/day) and the heat-killed bacteria group can reduce the inflammation index values (NLR and PLR), which can help improve inflammation after exercise and can be applied Due to diseases or symptoms caused by high NLR and PLR; another example shows that TWK10 has the effect of lowering body fat regardless of the form of live bacteria or non-activated bacteria, and the body fat of the live bacteria group decreases with increasing dose The more significant, it can be applied to diseases or symptoms caused by excessive body fat, and has the potential to improve chronic inflammation.
  • NLR and PLR inflammation index values
  • TWK10 for the first time can improve the symptoms of inflammation after exercise, which is different from other probiotics for inhibiting the diseases caused by inflammation, and provides rigorous human clinical data as evidence. In addition, it also provides the effect of reducing body fat. For athletes, it can be combined Anti-inflammation and body fat reduction after exercise, in all aspects to help improve body function.
  • the probiotic use of the present invention will contribute to the improvement of diseases and symptoms caused by inflammation or excessive body fat.
  • TWK10 can reduce body fat, and because fat releases inflammatory hormones, long-term use can reduce the body fat continuously under the original effect of improving inflammation after exercise, and thus achieve a synergistic effect to improve chronic inflammation.
  • Figure 1 The change of neutrophil/lymphocyte ratio after taking the probiotic composition of Lactobacillus plantarum TWK10 after exercise.
  • Figure 3 Changes in body fat after exercising with a probiotic composition containing Lactobacillus plantarum TWK10.
  • probiotics refers to microorganisms that can provide physiological health benefits to the user when properly administered.
  • the microorganisms have no toxicity, intestinal adhesion, and gain effect; colony forming unit (CFU) )
  • CFU colony forming unit
  • composition refers to a product that comes from mixing or combining more than one active ingredient;
  • carrier refers to a non-toxic compound or agent, which has the function of assisting cells or tissues to absorb drugs, and can be aromatics, buffers, binders, colorants, disintegrants Dissolving agents, thinners, emulsifiers, extenders, odor improvers, gelling agents, glidants, preservatives, skin penetration enhancers, cosolvents, stabilizers, suspending agents, sweeteners , Tonicity agents, viscosity enhancers, or any combination of the above;
  • the present invention is used to dissolve buffer solutions of salts and regulate pH value, including but not limited to phosphate buffer solution (PBS); the term “effective amount” refers to a sufficient amount of compound or medicine, which can be taken by the user Reduce one or more disease symptoms or physiological conditions after eating.
  • PBS phosphate buffer solution
  • the "effective amount” of treatment is a dose containing a compound provided by the present invention that can significantly reduce certain physiological indicators in the clinic;
  • maximum oxygen uptake (abbreviated as VO 2 max)” refers to a person in the sea On the plane, the highest value of oxygen that can be consumed or utilized by tissue cells when engaged in the most intense exercise;
  • endurance exercise ability test with fixed intensity and time refers to the intensity of 60% VO 2 max or more on the treadmill Or (or bicycle) exercise test for at least 30 minutes.
  • probiotic bacteria may be live bacteria or non-activated bacteria, the non-activated is treated by thermal lethal or other physical or chemical methods.
  • the Lactobacillus germ used in the present invention is Lactobacillus plantarum TWK10 (hereinafter referred to as TWK10), which was isolated by Dr. Cai Zongyou from the cabbage after fermentation in Taiwan, and is deposited at the China General Microbial Culture Collection Management Center (Address: Chaoyang, Beijing No. 3, No. 1, Yard Beichen West Road, Zip Code: 100101), its deposit number is CGMCC13008, and the deposit date is September 13, 2016; and the Taiwan Food Industry Development Institute, its deposit number is BCRC910734.
  • the classification of the embryo Lactobacillus TWK10 is named Lactobacillus plantarum.
  • the experiment uses a double-blind crossover design.
  • the subjects are between 20 and 40 years old. They need to take a two-week washing period, followed by six weeks of probiotics, and a fixed-intensity endurance exercise test on the beginning and end of the day. , Divided into 5 groups according to the maximum oxygen uptake (VO 2 max, mL/kg/min), each group of 16 people, and blood test to determine the changes of physiological factors; each group of probiotic composition taken three times a day, one capsule at a time, The dosage and strains given in each group are listed below:
  • Placebo group Placebo capsules without TWK10 (containing maltodextrin, lactose, microcrystalline ⁇ -cellulose), take 3 capsules daily, and take warm water after three meals.
  • TWK10-L Low-dose group-TWK10 live bacteria recommended double-dose group: 1 TWK10 capsule contains 1x10 10 CFU, 3 capsules per day, 3x10 10 CFU/day, after three meals with warm water .
  • TWK10-M Medium dose group-TWK10 live bacteria recommended 3 times dose group (TWK10-M): 1 TWK10 capsule containing 3x10 10 CFU, 3 capsules per day, 9x10 10 CFU/day, taken with warm water after three meals .
  • TWK10-H High-dose group-TWK10 live bacteria recommended 10-fold dose group: 1 TWK10 capsule contains 1x10 11 CFU, 3 capsules per day, 3x10 11 CFU/day, taken with warm water after three meals .
  • TWK10 heat-killing bacteria group (TWK10-HK): 1 TWK10 heat-killing bacteria capsule contains 1x10 11 CFU, which is taken 3 times a day, taken with warm water after three meals, 3x10 11 CFU/day.
  • NLR neutrophil/lymphocyte ratio
  • PLR platelet/lymphocyte
  • TWK10 in the high-, medium-, and low-dose and heat-killing bacteria groups could reduce NLR, of which the low-dose and heat-killing bacteria group decreased by about 30-40 times compared with the placebo group; It can be 1.2-1.8 times lower than the placebo group; in general, taking TWK10 dead bacteria can improve inflammation after exercise, and taking 3x10 10 CFU or more of TWK10 live bacteria daily can also improve inflammation after exercise. Dose, viable bacteria exceeding 9x10 10 CFU, such as 1x10 11 or 3x10 11 CFU, can achieve better results.
  • Example 2 Body fat lowering effect of Lactobacillus germ TWK10
  • TWK10 has the effect of reducing body fat in addition to improving the symptoms of inflammation after exercise.
  • the experiment adopts a double-blind crossover design.
  • the subjects are between 20 and 40 years old.
  • the probiotic composition is given for six weeks, and a fixed-strength is administered on the day of starting and ending Endurance exercise test, divided into 5 groups according to the maximum oxygen uptake, each group of 16 people, and determine the change in total body fat; each group of probiotic composition is taken three times a day, one capsule at a time, each group is given doses and strains and examples One is the same.
  • TWK10 has the effect of reducing inflammation and body fat after exercise.
  • TWK10 can reduce NLR or (and) PLR, NLR and PLR can also be used as diagnostic indicators for many cancers, disease metastases, and infections.
  • the cause, and excessive body fat is one of the causes of systemic chronic inflammation, chronic disease, cardiovascular disease, etc., so any use of the results of the present invention to improve related diseases or symptoms caused by NLR, PLR, or excessive body fat
  • the implementation of should be deemed to be included in the scope of the present invention.
  • probiotic composition of the present invention Due to the synergistic effect of reducing body fat and improving inflammation, long-term use of the probiotic composition of the present invention can not only reduce body fat, but also simultaneously reduce systemic chronic inflammation, achieving a double and additive body improvement effect.
  • the probiotic composition used in the present invention includes but is not limited to solution, emulsion, suspension, powder, tablet, pill, lozenge, troche, chewing gum ( chewing gums, capsules, and other dosage forms similar or applicable to the present invention.
  • the probiotic composition used in the present invention may optionally include other strains taught by the general knowledge of the technical field to which the present invention belongs, Lactobacillus sp., Lactococcus sp., Pediococcus (Pediococcus sp.), Leuconostoc sp., Enterococcus sp., Streptococcus sp., Bifidobacterium sp., yeasts, yeasts, Saccharomyces sp.).
  • the probiotic composition used in the present invention may be a food composition, which is further prepared into capsules, lozenges, beverages, powders or dairy products; the aforementioned composition used in the present invention may be further added with an edible material to prepare as A food product or health care product.
  • the edible material includes, but is not limited to: water, fluid dairy products, milk, concentrated milk; fermented dairy products, such as yogurt, yogurt, frozen yogurt, lactobacillus fermented beverages; Milk powder; ice cream; cheese; cheese; soy milk; fermented soy milk; vegetable juice; fruit juice; sports drink; dessert; jelly; candy; baby food; health food; animal feed; Chinese herbal medicines; dietary supplements, etc.
  • composition used in the present invention may be a dietary supplement and may be administered to the user in the following manner: mixed with a suitable drinkable liquid, such as water, yogurt, milk, or fruit juice; or may be mixed with solid or liquid food.
  • Dietary supplements can be in the form of lozenges, pills, capsules, lozenge, granules, powders, suspensions, sachets, pastilles, candies, sticks, syrups, and corresponding forms of administration, usually in units Dosage form, and manufactured in the usual way to prepare dietary supplements.
  • the effective dose varies according to the combination therapy.
  • Combination therapy may further include periodic therapy, which varies according to factors such as co-administered compositions, diseases, physiological abnormalities, physiological conditions, etc.

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Abstract

胚芽乳酸杆菌TWK10用于制备改善运动后发炎或降体脂的益生菌组合物的用途，该胚芽乳酸杆菌TWK10保藏于中国普通微生物菌种保藏管理中心，保藏编号为CGMCC 13008；前述益生菌组合物，包括药学上可接受的载剂、赋形剂、稀释剂，构成溶液、悬浮液、乳剂、粉末、锭剂、丸剂、糖浆、口含锭、片剂、口嚼胶、浓浆、和胶囊等剂型，并可进一步制成乳制品、豆乳制品、果汁饮料、糖果甜点、婴儿食品、健康食品、动物饲料、中草药组合物、膳食补充品等。

Description

[根据细则37.2由ISA制定的发明名称]　胚芽乳酸杆菌TWK10在制备抗运动后发炎或降体脂组合物的用途 技术领域
本发明为益生菌的应用领域，特别在于描述胚芽乳酸杆菌可用于抗运动后发炎或降体脂。
背景技术
益生菌因来源天然，对人体无危害，服用门坎低，且具有各种身体保健、调节功效，故极受健康食品的市场欢迎。其中，胚芽乳酸杆菌(Lactobacillus plantarum)为常见益生菌，是一种革兰氏阳性细菌，可见于发酵食物、或动物唾液中，其具有乳酸菌中最大的基因体，且变异繁多，故常因变异程度与适应条件不同，同种菌株间会表现出不同特征与功效。
先前报导指出胚芽乳酸杆菌具有多种医疗用途，包括保护呼吸道的病毒感染、以及混合于化疗药物后可增加癌细胞对药物的敏感性，另有多种食品添加物的用途，例如可用于防止生长于玉米的有毒真菌。
发炎指具有血管系统的活体组织对致炎因子及局部损伤所发生的防御性为主的反应，又可分为急性发炎和慢性发炎。急性发炎是生物体面对有害刺激的初步反应，更多的血浆和白血球(特别是粒细胞)从血液移往受损组织，进行一连串的连锁性的反应，当中牵涉局部的血管系统、免疫系统及受损组织内的各个细胞。慢性发炎导致发炎部位的细胞类型改变，令组织的毁灭与修复同步进行。此外，人类脂肪具有分泌发炎激素的能力，故体脂肪过高也是造成身体慢性发炎的因素。在一些特殊情形下，发炎甚至会引起人体自身免疫系统的过敏，进而攻击自身的组织及细胞，如：类风湿性关节炎和红斑 狼疮症等免疫系统过敏病症。长期慢性发炎可引起一系列疾病，会使得器官与组织逐渐失去功能，不但容易造成疲倦、老化、荷尔蒙失调，还会导致各种严重疾病，包括肝炎、肺炎、肾脏炎、自体免疫疾病、关节炎、糖尿病、阿兹海默症、心血管疾病、甚至癌症。
运动后造成肌肉纤维或是结缔组织上的伤害，会造成肌肉发炎，伴随而致的酸、疼痛、各种不适，及长期积累引发的慢性发炎，不仅会降低人们的运动意愿与生活质量，甚至会引发运动伤害、姿势改变。特别对于运动员而言，如何能有效降低运动后的发炎状况，更是提升练习及比赛效率、降低运动伤害的关键因素。
虽然市面上健康食品琳琅满目，但在台湾，却仅有五项经过台湾卫福部认证具有抗发炎功效(表1)，目的均在于调整过敏体质、抗过敏。然而，过敏的机制为过敏原的诱发、导致免疫球蛋白过度刺激肥大细胞的致敏化作用，与运动后发炎(其主要是肌腱或是韧带受到伤害)所对应的急性或慢性发炎不尽相同。目前，针对益生菌改善运动后发炎的应用上，仍待相关研究。
表1-卫生福利部食品药物管理署审核通过的抗发炎健康食品一览表
Figure PCTCN2018120722-appb-000001
先前相关文献公开，胚芽乳酸杆菌可用于改善抗发炎相关疾病，然而， 却未有可改善运动后发炎的益生菌的相关发表。
因此，如何能提供一种改善运动后发炎的益生菌新用途，是本发明所欲解决的问题。
发明内容
本发明提供一种胚芽乳酸杆菌TWK10用于制备改善运动后发炎的益生菌组合物的用途，该胚芽乳酸杆菌TWK10保藏于台湾食品工业发展研究所，保藏编号为BCRC910734，以及保藏于中国普通微生物菌种保藏管理中心(CGMCC)，保藏编号为CGMCC 13008。
为达到前述的发明目的，该胚芽乳酸杆菌TWK10改善运动后发炎是降低中性粒细胞/淋巴细胞比值(NLR)和血小板/淋巴细胞(PLR)比值。
为达前述发明的目的，该胚芽乳酸杆菌TWK10为活菌或不活化菌，不活化菌为经热致死或其他物理或化学处理。该胚芽乳酸杆菌TWK10为活菌时，剂量大于或等于3x10 10CFU/天；其中，剂量大于1x10 11CFU/天时，具有较佳的改善运动后发炎的功效。
本发明同时提供一改善运动后发炎的方法，为服用有效量的一益生菌组合物，该益生菌组合物包含保藏编号为CGMCC 13008的胚芽乳酸杆菌TWK10。
本发明进一步提供一种改善中性粒细胞/淋巴细胞比值和血小板/淋巴细胞比值过高所引发的疾病的方法，为服用有效量的一益生菌组合物，该益生菌组合物包含保藏编号为CGMCC 13008的胚芽乳酸杆菌TWK10。
本发明提供一种胚芽乳酸杆菌TWK10用于制备降体脂的益生菌组合物的用途，该胚芽乳酸杆菌TWK10保藏于台湾食品工业发展研究所，保藏编号为 BCRC910734，以及保藏于中国普通微生物菌种保藏管理中心(CGMCC)，保藏编号为CGMCC 13008。
为达前述发明的目的，该胚芽乳酸杆菌TWK10降体脂为降低全身体脂肪。
为达前述发明的目的，该益生菌组合物所包括的益生菌为活菌、不活化菌或是前述两者的组合；该胚芽乳酸杆菌TWK10服用剂量大于或等于3x10 10CFU/天。
本发明更提供一种降体脂的方法，系服用有效量的一益生菌组合物，该益生菌组合物包含保藏编号为CGMCC 13008的胚芽乳酸杆菌TWK10。
本发明进一步提供一种改善体脂过高所引发的疾病的方法，为服用有效量的一益生菌组合物，该益生菌组合物包含保藏编号为CGMCC 13008的胚芽乳酸杆菌TWK10。
前述的益生菌组合物，均可包括药学上可接受的载剂、赋形剂、稀释剂，剂型选自溶液、悬浮液、乳剂、粉末、锭剂、丸剂、糖浆、口含锭、片剂、口嚼胶、浓浆、和胶囊所构成的群组。
前述的益生菌组合物，可进一步制成流体乳品、浓缩牛奶、酸奶、酸乳、冷冻优格、乳杆菌发酵饮料、奶粉、冰淇淋、奶酪、干酪、豆浆、发酵豆浆、蔬果汁、果汁、运动饮料、甜点、果冻、糖果、婴儿食品、健康食品、动物饲料、中草药组合物、膳食补充品。
前述的益生菌组合物，包括下列益生菌群组其中之一或任意组合：乳杆菌属物种、乳球菌属物种、片球菌属物种、明串珠菌属物种、肠球菌属物种、链球菌属物种、双叉杆菌属物种、酵母菌。
本发明的一个实施例证明，TWK10活菌高剂量组(3x10 11CFU/天)及热致死菌组均可降低发炎指标的数值(NLR及PLR)，有助于改善运动后发炎，且可应用于NLR、PLR过高所导致的疾病或症状；另一实施例证明，TWK10不论是活菌或是非活化菌形态，均具有降体脂的功效，且活菌组随着剂量提升，体脂肪下降越显著，具有可应用于体脂过高所导致的疾病或症状，并具有改善慢性发炎的潜力。
本发明所提供的益生菌的新用途所具有的优势，如下所述：
(1)目标明确且新颖
首次提供TWK10可改善运动后发炎的症状，不同于其他益生菌针对抑制发炎引发的疾病，并提供严谨的人体临床数据佐证；此外，更同时提供降体脂的效果，对于运动员而言，可结合抗运动后发炎与降体脂，全方面地辅助增进身体机能。
(2)应用范围广泛
因发炎、体脂均为常见疾病或症状的成因或评估因子，故本发明益生菌用途将有助于发炎或体脂过高所引发的疾病、症状的改善。
(3)协同功效
TWK10可降体脂，而因脂肪会释放发炎激素，长期服用，可在原有的改善运动后发炎效果之下，因体脂持续降低，进而取得协同性地改善慢性发炎的功效。
附图说明
图1服用Lactobacillus plantarum TWK10的益生菌组合物，在运动后，中性粒细胞/淋巴细胞比值的变化。
图2服用含Lactobacillus plantarum TWK10的益生菌组合物，在运动后，血小板/淋巴细胞比值的变化。
图3服用含Lactobacillus plantarum TWK10的益生菌组合物，在运动后，体脂肪的变化。
具体实施方式
本发明将就下列实施例作进一步说明，然这些实施例仅为表现与叙述本发明的发明涵义与其精神，故所述实施例等用于例示说明，而不应作为限缩或被解释为实施本发明的限制或仅限于这些所述实施例或方式。
本发明说明书及申请专利范围中所述的所有技术性及科学用语，除非另有所定义，都依据以下叙述定义。其中单数用语“一”、“一个”、“该”，除非另有说明，皆可指代多于一个对象；除非另有说明，实施本发明采用的通常技术可包括临床检体采集与分析、高效液相层析法、蛋白质化学、生物化学、重组DNA技术、药理学技术；此外，用语“包含”、“包括”皆为开放式连接词；除非另有说明，本发明所用的材料皆市售易于取得。
用语“益生菌(probiotics)”，是指适当给予时可以给服用者提供生理健康的益处的微生物，该微生物具有无毒性、肠道附着力、及增益效果；菌落形成单位(CFU，colony forming unit)指单位体积中的细菌、霉菌、酵母等微生物的群落总数用语；用语“组合物”指代一产物，该产物来自混合或组合多于一种活性成分；“载剂(carrier)”、“赋形剂(vehicle)”，是指代不具毒性的化合物或药剂，其具有协助细胞或组织吸收药物的功能，可为芳香剂(aromatics)、缓冲剂、黏着剂(binders)、着色剂、崩解剂、稀释剂、乳化剂、增量剂(extenders)、气味改善剂、胶凝剂、滑动剂(glidants)、防腐剂、皮肤渗入促进剂、助溶剂、稳 定剂、悬浮剂、甜味剂、张力剂(tonicity agents)、黏度增强剂或上述的任意组合；用语“稀释剂”，是指代给予组合物前用以稀释的溶剂，可用于稳定化合物。本发明用于溶解盐类的缓冲溶液并调控酸碱值，包括但不限于磷酸盐缓冲液(PBS)；用语“有效量”，是指代化合物或药物的一足够量，可于服用者服食后减轻一项或多项疾病症状或生理状况。举例而言，治疗的“有效量」是包含一本发明提供化合物的可于临床上显著降低特定生理指标的剂量；用语“最大摄氧量(缩写为VO 2max)”，指一个人在海平面上，从事最激烈的运动时，组织细胞所能消耗或利用的氧的最高值；用语“固定强度与时间的耐力运动能力测试”，是指以60％VO 2max以上的强度在跑步机或(或脚踏车)进行运动测试至少30分钟。
以下实施仅为例示，本发明所使用的益生菌组合物，其中该益生菌可为活菌或不活化菌，该不活化经由热致死或其他物理或化学方式处理。
本发明所使用的胚芽乳酸杆菌菌株为Lactobacillus plantarum TWK10(以下简称TWK10)，为蔡宗佑博士于台湾发酵后的甘蓝菜中分离而得，保藏于中国普通微生物菌种保藏管理中心(地址：北京市朝阳区北辰西路1号院3号，邮政编码：100101)，其保藏编号为CGMCC13008，保藏日期为2016年9月13日；及台湾财团法人食品工业发展研究所，其保藏编号为BCRC910734。该胚芽乳酸杆菌TWK10的分类命名为植物乳杆菌(Lactobacillus plantarum)。
实施例一、胚芽乳酸杆菌TWK10的抗发炎功效
(一)实验设计与流程
实验采用双盲交叉设计，受试者介于20-40岁间，需经两周的洗涤期，接续六周益生菌的服用，并于服用起始与结束当天施予固定强度的耐力运动测 试，依最大摄氧量(VO 2max,mL/kg/min)分作5组，每组16人，并验血测定生理因子变化；各组益生菌组合物一天服用三次，一次一颗胶囊，每组给予剂量及菌株列举如下：
(1)安慰剂组：不含TWK10的安慰剂胶囊(内含麦芽糊精、乳糖、微结晶状α-纤维素)，每日食用3颗，三餐饭后配温开水服用。
(2)低剂量组-TWK10活菌建议1倍剂量组(TWK10-L)：1颗TWK10胶囊含有1x10 10CFU，每日食用3颗，3x10 10CFU/天，三餐饭后配温开水服用。
(3)中剂量组-TWK10活菌建议3倍剂量组(TWK10-M)：1颗TWK10胶囊含有3x10 10CFU，每日食用3颗，9x10 10CFU/天，三餐饭后配温开水服用。
(4)高剂量组-TWK10活菌建议10倍剂量组(TWK10-H)：1颗TWK10胶囊含有1x10 11CFU，每日食用3颗，3x10 11CFU/天，三餐饭后配温开水服用。
(5)TWK10热致死菌组(TWK10-HK)：1颗TWK10热致死菌胶囊含有1x10 11CFU，每日食用3颗，三餐饭后配温开水服用，3x10 11CFU/天。
(二)结果
分析六周后，每人在固定强度与时间(120分钟)的耐力运动测试后，各组的中性粒细胞/淋巴细胞比值(NLR)和血小板/淋巴细胞(PLR)比值的变化，两者代表体内淋巴系统的状态，均为检测体内发炎的指标，其一比值下降即表示发炎改善，甚至报导指出，可作为癌症转移或其他疾病的观测因子，结果如图1、图2，纵轴表示相对于试验起始所测得的NLR或PLR的变化量。
结果显示，高、中、低剂量和热致死菌组TWK10可降低NLR，其中低剂量和热致死菌组较安慰剂组下降约30-40倍；而PLR的部分，高剂量及热致死菌组可较安慰剂组下降1.2-1.8倍；总体而言，服用TWK10死菌可改善运动 后发炎，而每日服用3x10 10CFU以上TWK10活菌也具有改善运动后发炎功效，若每日服用更高剂量，即超过9x10 10CFU的活菌，例如1x10 11或3x10 11CFU，可达到更佳的效果。
实施例二、胚芽乳酸杆菌TWK10的降体脂功效
因脂肪细胞会分泌发炎激素，导致全身性发炎，故体脂高与慢性发炎也有密切的关系。接下来，将探讨TWK10除了可改善运动后发炎的症状外，是否也具有降体脂的功效。
(一)实验设计与流程同实施例一。
实验采双盲交叉设计，受试者介于20-40岁间，经两周的洗涤期及一周休息后，给予益生菌组合物六周，并于服用起始与结束当天施予固定强度的耐力运动测试，依最大摄氧量分作5组，每组16人，并测定全身体脂肪变化；各组益生菌组合物一天服用三次，一次一颗胶囊，每组给予剂量及菌株与实施例一相同。
(二)结果
服用益生菌组合物六周后，分析每人在固定强度与时间的耐力运动测试后，体脂肪的变化，结果如图3，纵轴表示服用结束相对于服用起始，所测得的体脂肪的变化百分比率。
结果显示，活菌及热致死菌组均可降低体脂肪，其中低剂量和热致死菌组较安慰剂组下降约0.5％，活菌组随着剂量提升，体脂肪下降越显著，高剂量组可令体脂肪下降达1.5％，表示TWK10不论是活菌或是非活化菌形态，均具有降体脂的功效。
综上所述，TWK10具有降低运动后发炎及降体脂功效，此外，因TWK10 可令NLR或(及)PLR下降，而NLR及PLR还可作为许多癌症、疾病的转移、感染的诊断指标与成因，且体脂过高是全身性的慢性发炎、慢性病、心血管疾病等的成因之一，故任何利用本发明结果以改善因NLR、PLR、或体脂过高所导致的相关疾病或症状的实施，均应视为包括于本发明请求范围内。
因降体脂与改善发炎具有协同效果，长期服用本发明益生菌组合物，不仅可降体脂，且可同步降低全身性慢性发炎，达到双重且加成的身体改善功效。
本发明所使用的益生菌组合物，其剂型包含但不限于溶液、乳剂、悬浮液、粉末、锭剂(tablet)、丸剂、口含锭(lozenge)、片剂(troche)、口嚼胶(chewing gum)、胶囊以及其他类似或适用本发明的剂型。
本发明所使用的益生菌组合物，可选择性地包括本发明所属技术领域的通常知识所教导的其他菌种，乳杆菌属(Lactobacillus sp.)、乳球菌属(Lactococcus sp.)、片球菌属(Pediococcus sp.)、明串珠菌属(Leuconostoc sp.)、肠球菌属(Enterococcus sp.)、链球菌属(Streptococcus sp.)、双叉杆菌属(Bifidobacterium sp.)、酵母菌(yeasts,Saccharomyces sp.)。
本发明所使用的益生菌组合物可为食品组合物，进一步制成胶囊、锭剂、饮料、粉末或乳制品；本发明所使用的前述组合物，可进一步添加一可食性材料，以制备为一种食品产品或保健产品。其中该可食性材料包含，但不限于：水、流体乳品、牛奶、浓缩牛奶；发酵乳品，诸如酸奶(yogurt)、酸乳(sour milk)、冷冻优格(frozen yogurt)、乳杆菌发酵饮料；奶粉；冰淇淋；奶酪；干酪；豆奶；发酵豆奶；蔬果汁；果汁；运动饮料；甜点；果冻；糖果；婴儿食品；健康食品；动物饲料；中草药材；膳食补充品等。
本发明所使用的前述组合物可为膳食补充品，可以下列方式给予服用者：与合适的可饮用液体混合，例如水、酸奶酪、牛奶或果汁；或可与固体或液体食品进行混合。膳食补充品的形式可为锭剂、丸剂、胶囊、药锭糖(lozenge)、颗粒、粉剂、悬浮剂、小药囊、软锭剂、糖果、棒、糖浆及相应的给予形式，通常为单位剂量的形式，并以制备膳食补充品的常规方法制造。
在本发明实施例中，有效剂量依据组合治疗而变动。组合治疗可进一步包括周期性治疗，依据共同给予的组合物、疾病、生理异常、生理状况等因子而变动。
以上所述的实施例仅为说明本发明的技术思想及特点，其目的使本领域技术人员能够了解本发明的内容并据以实施，但不能以此限定本发明的专利范围，即凡依本发明所揭示的精神所作的均等变化或修饰，仍应涵盖在本发明的专利范围内。

Claims (21)

  1. 一种胚芽乳酸杆菌TWK10用于制备改善运动后发炎的益生菌组合物的用途，所述胚芽乳酸杆菌TWK10保藏于中国普通微生物菌种保藏管理中心，保藏编号为CGMCC 13008。
  2. 如权利要求1所述的用途，所述改善运动后发炎为降低中性粒细胞/淋巴细胞比值或血小板/淋巴细胞比值。
  3. 如权利要求1所述的用途，所述胚芽乳酸杆菌TWK10为活菌或不活化菌，所述不活化菌经物理或化学处理，其中，物理处理包含热致死。
  4. 如权利要求3所述的用途，所述胚芽乳酸杆菌TWK10为活菌时，剂量大于或等于3x10 10CFU/天。
  5. 如权利要求4所述的用途，剂量大于1x10 11CFU/天。
  6. 如权利要求1所述的用途，所述益生菌组合物包括药学上可接受的载剂、赋形剂、稀释剂。
  7. 如权利要求1所述的用途，所述益生菌组合物的剂型选自溶液、悬浮液、乳剂、粉末、锭剂、丸剂、糖浆、口含锭、片剂、口嚼胶、浓浆、和胶囊所构成的群组。
  8. 如权利要求1所述的用途，所述益生菌组合物进一步制成流体乳品、浓缩牛奶、酸奶、酸乳、冷冻优格、乳杆菌发酵饮料、奶粉、冰淇淋、奶酪、干酪、豆浆、发酵豆浆、蔬果汁、果汁、运动饮料、甜点、果冻、糖果、婴儿食品、健康食品、动物饲料、中草药组合物、膳食补充品。
  9. 如权利要求1所述的用途，所述益生菌组合物包括下列益生菌群组其中之一或任意的组合：乳杆菌属、乳球菌属、片球菌属、明串珠菌属、肠球菌属、链球菌属、双叉杆菌属、酵母菌。
  10. 一种用于制备改善运动后发炎的醫藥或食品组合物的用途，包含保藏编号为CGMCC 13008的胚芽乳酸杆菌TWK10。
  11. 一种用于制备改善中性粒细胞/淋巴细胞比值和血小板/淋巴细胞比值过高所引发的疾病或症状的益生菌组合物的用途，包含保藏编号为CGMCC 13008的胚芽乳酸杆菌TWK10。
  12. 一种胚芽乳酸杆菌用于制备降体脂的益生菌组合物的用途，所述胚芽乳酸杆菌是保藏编号为CGMCC 13008的胚芽乳酸杆菌TWK10。
  13. 如权利要求12所述的用途，所述降体脂为降低全身体脂肪。
  14. 如权利要求12所述的用途，所述益生菌组合物所包括的益生菌为活菌、不活化菌或是前述两者的组合。
  15. 如权利要求12所述的用途，所述胚芽乳酸杆菌TWK10服用剂量大于或等于3x10 10CFU/天。
  16. 如权利要求12所述的用途，所述益生菌组合物包括药学上可接受的载剂、赋形剂、稀释剂。
  17. 如权利要求12所述的用途，所述益生菌组合物的剂型选自溶液、悬浮液、乳剂、粉末、锭剂、丸剂、糖浆、口含锭、片剂、口嚼胶、浓浆、和胶囊所构成的群组。
  18. 如权利要求12所述的用途，所述益生菌组合物进一步制成流体乳品、浓缩牛奶、酸奶、酸乳、冷冻优格、乳杆菌发酵饮料、奶粉、冰淇淋、奶 酪、干酪、豆浆、发酵豆浆、蔬果汁、果汁、运动饮料、甜点、果冻、糖果、婴儿食品、健康食品、动物饲料、中草药组合物、膳食补充品。
  19. 如权利要求12所述的用途，所述益生菌组合物包括下列益生菌群组其中之一或任意的组合：乳杆菌属物种、链球菌属物种、双叉杆菌属物种、酵母菌。
  20. 一种用于制备降体脂的醫藥或食品组合物的用途，包含保藏编号为CGMCC 13008的胚芽乳酸杆菌TWK10。
  21. 一种用于制备改善体脂过高所引发的疾病或症状的益生菌组合物的用途，包含保藏编号为CGMCC 13008的胚芽乳酸杆菌TWK10。
PCT/CN2018/120722 2018-12-12 2018-12-12 胚芽乳酸杆菌twk10在制备抗运动后发炎或降体脂组合物的用途 WO2020118576A1 (zh)

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