US20090324733A1 - Composition of Whey Growth Factor Extract for Reducing Muscle Inflammation - Google Patents

Composition of Whey Growth Factor Extract for Reducing Muscle Inflammation Download PDF

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US20090324733A1
US20090324733A1 US11/991,696 US99169606A US2009324733A1 US 20090324733 A1 US20090324733 A1 US 20090324733A1 US 99169606 A US99169606 A US 99169606A US 2009324733 A1 US2009324733 A1 US 2009324733A1
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composition
whey
growth factor
subject
factor extract
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Michelle Rowney
David Cameron-Smith
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Murray Goulburn Co Opeartive Co Ltd
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Murray Goulburn Co Opeartive Co Ltd
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Publication of US20090324733A1 publication Critical patent/US20090324733A1/en
Priority to US13/602,864 priority Critical patent/US9480717B2/en
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    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A61K35/20Milk; Whey; Colostrum
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23CDAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
    • A23C21/00Whey; Whey preparations
    • A23C21/06Mixtures of whey with milk products or milk components
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23JPROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
    • A23J1/00Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites
    • A23J1/20Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from milk, e.g. casein; from whey
    • A23J1/205Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from milk, e.g. casein; from whey from whey, e.g. lactalbumine
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23LFOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/17Amino acids, peptides or proteins
    • A23L33/19Dairy proteins
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23LFOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/40Complete food formulations for specific consumer groups or specific purposes, e.g. infant formula
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00Medicinal preparations containing peptides
    • A61K38/16Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/1703Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
    • A61K38/1709Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00Medicinal preparations containing peptides
    • A61K38/16Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/18Growth factors; Growth regulators
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P21/00Drugs for disorders of the muscular or neuromuscular system
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23VINDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00Food compositions, function of food ingredients or processes for food or foodstuffs
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23VINDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2300/00Processes
    • A23V2300/30Ion-exchange

Definitions

  • the invention relates to the production of compositions containing milk products for use as a nutritional supplement. More specifically, it relates to compositions containing whey growth factor extract, as well as their use in methods for reducing muscle inflammation in individuals undertaking resistance exercise training.
  • Nutritional supplements are not intended to provide all the nutrients necessary for a complete diet, but instead are generally intended to complement the dietary intake such that it becomes more nutritionally complete. It is recognised that vitamins, minerals and other substances found in such supplements play important physiological roles and that a deficiency of certain vitamins, minerals and/or other components of supplements has been linked to development of certain diseases, a decrease in general health or lower performance in athletes.
  • nutritional supplements are known to enhance a variety of physiological states, under various conditions.
  • targets for nutritional supplements for example sick patients, convalescing patients, elderly persons and persons undergoing strenuous exercise regimes who wish to improve their performance and/or recovery from such exercise.
  • the nutritional requirements of bodybuilders and persons engaged in strenuous physical exercise are quite particular, whether to decrease body fat and increase lean muscle mass, and or to improve recovery from the strenuous exercise. Recovery includes the ability to overcome exercise-induced inflammation.
  • Inflammation is a localised response to injury or destruction of tissues. It is characterised in the acute form by pain, heat, redness, swelling, and loss of function. Accordingly, reduced muscle inflammation includes, but is not limited to a reduction in any one or more of the aforementioned symptoms in muscle.
  • the inflammatory response antagonises muscle protein synthesis and contributes to much of the physical discomfort experienced by persons 1 to 2 days after engaging in strenuous exercise, including resistance exercise training (Macintyre D L, et al., Sports Med. 1995; 20(1):24-40; MacIntyre D L, et al., Eur J Appl Physiol. 2001; 84(3):180-6). If the inflammation is severe, anti-inflammatory drugs such as corticosteroids may be prescribed. However, the use of such drugs can have deleterious side-effects and, in many cases, is prohibited for athletes in competition.
  • Weight-bearing or resistance training exercise elicits a wide-range of responses in the activated muscle.
  • Modulation of the expression of various genes is an essential process in regulating the sequence of cellular events needed to activate stem cells that are located within the muscle bed (satellite cells) (Anderson & Wozniak, Can J Physiol Pharmacol. 2004; 82(5):300-10) to rapidly proliferate, before maturing and ultimately fusing with existing muscle fibres or joining together to create new muscle fibres.
  • Regulators of the activation of satellite cells include Syndecan-3 (a transmembrane heparin sulphate proteoglycan essential for satellite cell proliferation), and Pax-7 (a protein that is essential for satellite cell activation and necessary for muscle tissue repair) (Seale et al., Dev Biol.
  • a natural food product such as a product derived from milk, could be obtained having beneficial effects on exercise recovery it would be a readily available and safe therapeutic supplement.
  • Colostrum is the pre-milk produced immediately after birth before the breast secretions stabilize into milk.
  • Prime colostrum from cows is obtained within the first six hours after calving and contains four times the protein found in milk from the same cow obtained 48 hours later.
  • the immunoglobulin fraction of colostrum is loaded with antibodies, lactoferrin and immune enhancers. Lactoferrin is thought to enhance the anti-inflammatory effect of colostrum, and is the subject of U.S. Pat. No. 6,475,511 (Gohlke MB). Immunoglobulins are thought to act locally on immune function in the gut.
  • Protein supplementation has been used widely by persons undertaking resistance exercise training to promote muscle protein synthesis in order to repair muscle tissue and facilitate muscle growth.
  • the nutritional supplementation may be provided in the form of a drink or food and includes protein powders to be mixed with liquid for use, nutritional bars and snack foods, tablets, capsules and other preparations.
  • Suitable protein sources commercially available include hydrolysed milk proteins, caseinates, soy protein isolates and milk protein concentrates prepared from ultra-filtrated skim milk.
  • Nutritional supplements which are based on other protein sources, such as whey protein, are also available and can be provided in the form of fruit juices, but are thought to be inadequate because they do not also provide a lipid source (WO 02/15720).
  • it has been considered that some milk-derived proteins are not readily absorbed by the gut, or do not survive the harsh environment of the digestive system to have a therapeutic effect.
  • Whey growth factor extract is one such milk product which was thought to be susceptible to at least partial loss of biological activity once ingested due to fragmentation of molecules such as growth factors. Moreover, by virtue of the process for its isolation, WGFE contains only low levels of immunoglobulins which, as described above, are thought to be important anti-inflammatory factors.
  • WPI Whey protein isolate
  • MPI milk protein isolate
  • WPI WPI
  • whey protein concentrate in combination with the amino acids glutamine, leucine, isoleucine and valine to improve muscular fatigue in a rat model is the subject of WO 2004/049830 A1 (Tsuchita H et al.). The authors measured Tyrosine release from soleus muscle as the indicator of fatigue.
  • whey growth factor extract altered the expression of genes involved in exercise-induced muscle inflammation.
  • trained athletes administered whey growth factor extract reported a reduction in post-exercise soreness.
  • whey growth factor extract may be isolated from normal bulk milk products without a requirement to maintain cows in a hyperimmune state or to harvest colostrum which is in limited supply relative to other milk products.
  • the invention relates to a WGFE composition and method of using the composition which enables persons undergoing resistance exercise training to reduce their level of muscle inflammation over that obtained with compositions of the prior art.
  • a skeletal muscle inflammation-reducing composition comprising whey growth factor extract, isolated from a milk product by cation exchange chromatography.
  • a muscle anti-inflammatory composition comprising whey growth factor extract, isolated from a milk product by a process comprising the steps;
  • a muscle anti-inflammatory composition comprising whey growth factor extract, isolated from a milk product by a process comprising the steps;
  • whey growth factor extract is isolated from a milk product selected from whole milk, cheese whey, rennet casein whey, acid casein whey, or concentrates thereof, or skim milk.
  • composition according to the above when used as a muscle anti-inflammatory agent when used as a muscle anti-inflammatory agent.
  • a method of reducing skeletal muscle inflammation in subjects undertaking resistance exercise training comprising administering to the subjects an efficacious amount of a composition comprising whey growth factor extract.
  • a method of reducing skeletal muscle inflammation in subjects undertaking resistance exercise training comprising administering to the subjects an efficacious amount of a composition comprising whey growth factor extract, isolated from a milk product by a process comprising the steps;
  • a method of reducing skeletal muscle inflammation in subjects undertaking resistance exercise training comprising administering to the subjects an efficacious amount of a composition comprising whey growth factor extract, isolated from a milk product by a process comprising the steps;
  • a method of reducing skeletal muscle inflammation in subjects undertaking resistance exercise training wherein the amount of whey growth factor extract administered, per daily dose, is at least 5 mg/kg body weight to 12.5 mg/kg body weight.
  • the daily dose of whey growth factor extract is at least 25 mg/kg body weight.
  • whey growth factor extract isolated from a milk product by cation exchange chromatography, for the production of a medicament for the treatment of a subject in need of reduced skeletal muscle inflammation.
  • whey growth factor extract isolated from a milk product by a process described above, for the production of a medicament for the treatment of a subject in need of reduced skeletal muscle inflammation.
  • a skeletal muscle inflammation-reducing composition comprising whey growth factor extract, isolated from a milk product by cation exchange chromatography, and an additional protein source.
  • the additional protein source is whey protein, preferably whey protein isolate (WPI), more preferably whey protein isolate wherein the whey protein isolate comprises:
  • a method of reducing skeletal muscle inflammation in a subject undertaking resistance exercise training comprising administering to the subject an efficacious amount of a composition comprising whey growth factor extract and an additional protein source, such as whey, preferably WPI.
  • a method of reducing muscle inflammation in a subject undertaking resistance exercise training comprising administering to the subject a composition comprising an effective amount of whey growth factor extract, isolated from a milk product by cation exchange chromatography, and an additional protein source taken separately.
  • the amount of additional protein source administered, per daily dose is at least 225 mg/kg body weight (dry weight) and preferably at least 435 mg/kg body weight (dry weight).
  • the administration of the composition of the invention is once per two or three days up to at least once per day, preferably before and/or immediately after resistance exercise training, more preferably immediately after resistance exercise training, most preferably between 20 minutes and two hours after exercise.
  • whey growth factor extract isolated from a milk product by cation exchange chromatography, and an additional protein source for the production of a medicament for the treatment of a subject in need of reduced skeletal muscle inflammation.
  • a food or drink comprising the composition of the invention for use in a method to reduce skeletal muscle inflammation in a subject undertaking resistance exercise training.
  • composition of the invention for the manufacture of a food or drink to reduce skeletal muscle inflammation in a subject undertaking resistance exercise training.
  • FIG. 1 Fold change in Myo D, myogenin, and myostatin gene (mRNA) expression in the vastus lateralis muscle of adult subjects, at rest and following 3 hours of resistance exercise training performed at the start of the trial [pre-training] and following 12 weeks of treatment [post-training] with 20 g WPI (Group A), 1 g WGFE+20 g WPI (Group B1), or 2 g WGFE+20 g WPI (Group B2). Results are presented as mean ⁇ SEM.
  • FIG. 2 Fold change in Pax 7 and Syndecan 3 gene (mRNA) expression in the vastus lateralis muscle of adult subjects, at rest and following 3 hours of resistance exercise training performed at the start of the trial [pre-training] and following 12 weeks of treatment [post-training] with 20 g WPI (Group A),1 g WGFE+20 g WPI (Group B1), or 2 g WGFE+20 g WPI (Group B2). Results are presented as mean ⁇ SEM.
  • FIG. 3 Percentage of Pax 7 positive nuclei in tissue sections from the vastus lateralis muscle of adult subjects, following 3 hours of resistance exercise training performed at the start of the trial [pre-training] and following 12 weeks of treatment [post-training] with 20 g WPI (Group A), 1 g WGFE+20 g WPI (Group B1), or 2 g WGFE+20 g WPI (Group B2) as analysed by immunohistochemical staining.
  • FIG. 4 Fold change in IGF-1 Binding Protein 4 and 5 gene (mRNA) expression in the vastus lateralis muscle of adult subjects, at rest and following 3 hours of resistance exercise training performed at the start of the trial [pre-training] and following 12 weeks of treatment [post-training] with 20 g WPI (Group A), 1 g WGFE+20 g WPI (Group B1), or 2 g WGFE+20 g WPI (Group B2). Results are presented as mean ⁇ SEM.
  • FIG. 5 Fold change in Jun B and c-fos gene (mRNA) expression in the vastus lateralis muscle of adult subjects, at rest and following 3 hours of resistance exercise training performed at the start of the trial [pre-training] and following 12 weeks of treatment [post-training] with 20 g WPI (Group A), 1 g WGFE+20 g WPI (Group B1), or 2 g WGFE+20 g WPI (Group B2). Results are presented as mean ⁇ SEM.
  • FIG. 6 Fold change in CCL2, 4 and 7 gene (mRNA) expression in the vastus lateralis muscle of adult subjects, at rest and following 3 hours of resistance exercise training performed at the start of the trial [pre-training] and following 12 weeks of treatment [post-training] with 20 g WPI (Group A), 1 g WGFE+20 g WPI (Group B1), or 2 g WGFE+20 g WPI (Group B2). Results are presented as mean ⁇ SEM.
  • FIG. 7 TNF-alpha production (ng/ml) by RAW cells exposed to lipopolysaccharide (LPS) and cultured in the presence of colostrum (sample 2BC), WPI (sample 4BC) at 100 mg/ml, WGFE (sample 6BC) at 100 mg/ml, various milk fractions, or LPS alone (control). WGFE and LPS were also added to the culture simultaneously (sample WGFE+LPS). The values plotted represent the arithmetic mean ⁇ SEM of the average response obtained in 3 independent experiments.
  • FIG. 8 Comparison of TNF-alpha production (ng/ml) by RAW cells exposed to lipopolysaccharide (LPS) and cultured in the presence of 100 mg/ml WPI or 100 mg/ml WGFE.
  • LPS lipopolysaccharide
  • the invention relates to a composition containing WGFE and uses thereof to enable a subject undergoing resistance exercise training to reduce their level of muscle inflammation over that obtained with compositions of the prior art.
  • a skeletal muscle inflammation-reducing composition comprising whey growth factor extract, isolated from a milk product by cation exchange chromatography.
  • the whey growth factor extract for use in the invention may be isolated from milk, skim milk, milk derivatives, whey, colostrum, and colostrum derivatives by, for example, the method described in Australian Patent No. 645589 (PCT/AU91/00303) which is incorporated herein by reference. This method essentially relies on strong cation exchange chromatography to selectively extract basic proteins from the starting material to constitute whey growth factor extract.
  • a preferred method of producing WGFE for use in the invention is to use a column packed with SP (sulphopropyl) Sepharose To the column a flow of a dairy product, preferably skimmed milk, is applied until the volume of milk applied is up to 1000 times the volume of the resin packed into the column. The milk remaining in the column is removed with a buffer of low ionic strength ( ⁇ 0.008M NaCl or equivalent) for 10 min. The WGFE fraction is eluted from the column with a buffer containing sodium ions equivalent to 0.4-0.5M NaCl (though other cations would be suitable), most preferably 0.4M NaCl.
  • the mobile phase may have a pH within a broad range, such as 4.5-9.0, preferably 5.5-7.5, most preferably about 6.5. At the upper and lower limits both protein stability and the ability of proteins to bind to the cation exchange resin become influenced. A pH in the range 5.5-7.5 provides the highest WGFE yields.
  • the type of cation exchange resin suitable for adsorption of the WGFE components may include resins such as Sepharose cation exchange resin beads.
  • resins such as Sepharose cation exchange resin beads.
  • SP Sepharose Big Beads and CM Sepharose beads products of GE Healthcare which contain sulfopropyl functional groups and carboxymethyl groups, respectively, are suitable.
  • the size of the cation exchange resin beads is preferably in the range from 45-300 ⁇ m. Both SP Sepharose beads in the range 45-165 ⁇ m and in the range 100-300 ⁇ m are suitable for WGFE purification according to the invention.
  • One of the further treatments to which the WGFE fraction can be subjected is desalting by, for example, dialysis or ultrafiltration.
  • whey growth factor extract is isolated from whey or skim milk.
  • the whey used as starting material may be cheese whey, rennet casein whey, acid casein whey, or concentrates thereof.
  • the amounts of whey growth factor extract and protein source to use according to the invention are to be sufficient for reduced muscle inflammation or to have a therapeutic effect.
  • a dosage regime wherein the amount of whey growth factor extract administered, per daily dose, is at least 5 mg/kg body weight to 12.5 mg/kg body weight.
  • the daily dose of whey growth factor extract is at least 25 mg/kg body weight.
  • a method of reducing skeletal muscle inflammation in subjects undertaking resistance exercise training comprising administering to the subjects an efficacious amount of a composition comprising whey growth factor extract.
  • composition includes an additional protein source, which may be any protein source suitable for consumption such as WPI, post-exercise inflammation is reduced compared to subjects administered a protein source alone.
  • the protein source may be obtained from whole milk, preferably whey protein and more preferably whey protein isolate (WPI).
  • WPI whey protein isolate
  • One such whey protein isolate is available commercially under the trade name NatraProTM by Murray Goulburn Co-Op Company Ltd.
  • a typical composition of NatraProTM WPI includes:
  • the composition according to the invention also contains a protein source and, in a dosage regime, the amount of additional protein source per daily dose administered is at least 225 mg/kg body weight (dry weight) and preferably at least 435 mg/kg body weight (dry weight).
  • the additional protein source could be formulated with the WGFE composition or formulated to be administered separately to the WGFE composition.
  • whey growth factor extract for the production of a medicament for the treatment of a subject in need of reduced skeletal muscle inflammation.
  • athletes may require resistance exercise training to build up their muscle strength to enhance their performance, or to overcome a debilitating injury which has caused muscle wasting.
  • the muscle inflammation may be caused by, for example, a ‘corked’ thigh, or a torn muscle.
  • a medicament comprising the composition of the invention may further assist in the subject recovering more rapidly to return to their training regime and/or to compete. Muscle inflammation may also occur in those with a propensity for bruising, such as the elderly.
  • a method of reducing skeletal muscle inflammation in subjects undertaking resistance exercise training comprising administering to the subjects an efficacious amount of a composition comprising whey growth factor extract and an additional protein source.
  • the administration of the composition of the invention may be on the days of training or on the days of training and/or other days, providing the regimen of administration results in reduced skeletal muscle inflammation.
  • administration is on the days of exercise and more preferably administration occurs either just before and/or immediately after the exercise. More preferably, administration occurs between 20 minutes and 2 hours after exercise.
  • a method as described above wherein the administration is immediately after the exercise.
  • Subjects administered 1 g WGFE plus 20 g WPI, or 2 g WGFE plus 20 g WPI immediately after resistance exercise training showed increased modulation of the expression of genes involved in inflammation compared to subjects administered WPI alone (see Examples and Results).
  • subjects administered WGFE reported reduced post-exercise soreness compared to those administered WPI, supporting a role for WGFE in reducing post-exercise muscle inflammation.
  • composition of the invention may be used to reduce muscle inflammation in non-human mammals such as horses, greyhounds and others in which reduced muscle inflammation is desirable.
  • a food or drink comprising the composition of the invention for use in a method to reduce skeletal muscle inflammation in subjects undertaking resistance exercise training.
  • composition of the invention for the manufacture of a food or drink to reduce skeletal muscle inflammation in subjects undertaking resistance exercise training.
  • WPI typically contains about 90% w/v protein; therefore 20 g of WPI contains about 18 g of protein as a nutritional source.
  • Whey growth factor extract typically contains about 85% w/v protein; therefore 2 g of whey growth factor extract contains about 1.7 g of protein as a nutritional source.
  • Whey growth factor extract was prepared according to the method broadly described in Australian Patent No. 645589 (PCT/AU91/00303), more specifically described supra.
  • Each whey protein formulation contained an artificial sweetener (NutrasweetTM, Nutrasweet Company, USA). Whey protein formulations were consumed immediately after each exercise session, with each subject completing three supervised exercise sessions per week.
  • a typical composition of NatraProTM WPI includes:
  • Muscle samples were collected from the vastus lateralis muscle of the right leg using the percutaneous needle biopsy technique. Excised muscle tissue was visually inspected, dissected free of any fat or connective tissue and blotted to remove excess blood and immediately frozen in liquid nitrogen for subsequent analysis. A portion of the muscle tissue was mounted in an aqueous mounting medium and frozen in isopentane cooled in liquid nitrogen for subsequent immunohistochemical analysis.
  • Results are presented as mean ⁇ SEM and significance calculated by two-way ANOVA using Bonforoni post hoc tests. No significant differences in age, weight, height, or BMI values were observed both pre and post training.
  • FIG. 1 The expression of Pax 7 and Syndecan 3, two markers of satellite cell recruitment and proliferation, was increased 2-fold and 7-fold, respectively, following 12 weeks of training and is supported by protein expression data ( FIGS. 2 and 3 ).
  • the expression of IGFBP4 and IGFBP5 ( FIG. 4 ) were markedly increased in group B1 at 12 weeks in response to acute exercise.
  • the normal post-exercise up-regulation of the early response genes jun-B and c-fos which are linked to muscle inflammation (Chen Y W et al., J. App. Physiol.
  • Whey growth factor extract was prepared as for Example 1.
  • the data supports that WGFE reduces the inflammation response in muscle following resistance exercise training.
  • In vitro data suggests that WGFE reduces inflammation by inhibiting TNF-alpha expression.
  • the data also supports that WGFE in combination with an additional protein source such as WPI decreases the post-exercise inflammatory response to a greater extent than observed when WPI, a protein source known to be used by subjects undertaking resistance exercise training, is administered alone.

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US20130195990A1 (en) 2013-08-01
CN101282736B (zh) 2011-12-28
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CN101282736A (zh) 2008-10-08
JP2013166760A (ja) 2013-08-29
EP1940426A1 (en) 2008-07-09
JP5841556B2 (ja) 2016-01-13
KR20080050478A (ko) 2008-06-05
WO2007028211A1 (en) 2007-03-15
CA2621739A1 (en) 2007-03-15
US9480717B2 (en) 2016-11-01
NZ566691A (en) 2011-06-30
JP2009507045A (ja) 2009-02-19

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