WO2009013723A2

WO2009013723A2 - Ethyl pyruvate alone or in combination with mi cro- clustered water and use of the same

Info

Publication number: WO2009013723A2
Application number: PCT/IB2008/052993
Authority: WO
Inventors: Ranya Alexander; Lee H Lorenzen
Original assignee: Donald, Heather June
Priority date: 2007-07-26
Filing date: 2008-07-25
Publication date: 2009-01-29
Also published as: WO2009013723A3

Abstract

The invention relates to the use of a pyruvate compound in the form of ethyl pyruvate in the preparation of an oral supplement or topical preparation for use in enhancing cell metabolism and absorption of medications in a human or animal body. The invention also includes a composition comprising ethyl pyruvate and micro-clustered water.

Description

ETHYL PYRUVATE AND USE OF THE SAME

BACKGROUND TO THE INVENTION

Pyruvate is the anionic structure of pyruvic acid, which is a naturally occurring acid that contains three carbons. Pyruvate plays an important role in the glycolytic and pyruvate dehydrogenase pathways in a human or animal body, helping the body to convert food into usable energy. It is naturally produced in the human body, but can also be supplemented through intake of fruits and vegetables that contain high levels of pyruvate, such as apples, dark beers and red wine. An average-sized person should take in between 10Omg and 2g of pyruvate daily.

Current studies are showing that pyruvate in supraphysiological amounts could possibly help the cardiovascular system as an isotropic. Supraphysiological levels of pyruvate could also have bariatric and ergogenic functions.

Pyruvate may also be effective in lowering body mass. One study gave 22g - 44g a day of pyruvate to subjects who were trying to lose weight using a low-fat diet. They gave other subjects a placebo. The subjects who took pyruvate iost more weight more quickly and had increased lean body mass.

However, pyruvic acid is a strong and unstable acid and on standing in solution, pyruvic acid and its salts at various pH values, including in the physiological range, are known to form both a stable hydrate and a dimer (para-pyruvate), neither of which react with reactive oxygen species as antioxidants and both of which are known inhibitors of pyruvate utilization as a metabolic fuel, thereby eliminating any of the beneficial effects which might have occurred from pyruvate administration. Furthermore, it has been recognized that traditional pharmacological pyruvate compounds, such as salts of pyruvic acid, are not particularly physiologically suitable. For example, these compounds lead to the accumulation of large concentrations of ions (e.g. calcium or sodium) in a patient's body fluids. Similarly, amino acid compounds containing pyruvate can lead to excessive nitrogen loads.

Ethyl pyruvate (ethyl-2-oxopropanoate) appears to function more effectively in reactions than other pyruvate forms and intravenous use of ethyl pyruvate for cellular energy is well known from the prior art. U.S. Patent No. 5,876,916 pertains to the intravenous use of pyruvate thiolesters and polyol esters for the treatment or prevention of oxidative injury, diabetic effects, cholesterol levels, injured organs, ethanoi intoxication or as a foodstuff. U.S. Patent Nos. 5,633,285; 5,648,380; 5,652,274; and 5,658,957 disclose various compositions, salts, prodrugs and derivatives of pyruvate in mixtures with other antioxidants, fatty acids as antiinflammatory and immuno-stimulating wound healing compositions.

There is, however, a need to provide a potable, easily ingestible form of ethyl pyruvate that will allow for the same cellular and organ benefits that are derived from intravenous ethyl pyruvate.

SUMMARY OF THE INVENTION

According to one aspect of the invention, there is provided a composition comprising pyruvate compound in the form of ethyt pyruvate and micro- clustered water.

The ethyl pyruvate may be in liquid or powder form.

The micro-clustered water may be micro-clustered water produced according to the method as claimed in U.S. Patent No. 6,033,678, which is incorporated herein by reference. For purposes of this specification, "micro-clustered" water (or "terahertz terahertz treated water") will be interpreted to mean water which has been treated to have a more ordered and stable clustering of water molecules when compared to, for example, with "standard" water.

In one form of the invention, the composition may be in the form of an oral supplement. in particular, the composition may be included in a consumable beverage or food stuff, such as an energy or diet bar. More particularly, the composition may be introduced into soft drinks, fruit juices, and coffee- and tea-based drinks. The tea-based drinks may include Camellia sinensis var. sinensis and Camellia sinensis var. assamica, and includes a blend of these teas.

The ethyl pyruvate may be imprinted or infused with far infrared or other electromagnetic radiation before being incorporated into a consumable beverage or food stuff.

Alternatively, the composition may orally be introduced into a human or animal body in the form of a tablet or capsule. In this form of the invention, the ethyl pyruvate may be a liquid or powder and may be included in a granulation process together with other solid nutrients for use in the tablet or capsule.

In another form of the invention, the composition may be in the form a topical preparation, such as an ocular, mucous membrane, otic or transdermal preparation.

The invention includes pyruvate compound in the form of ethyl pyruvate for use in enhancing cell metabolism and absorption of medications in a human or animal body.

According to another aspect of the invention there is provided the use of a pyruvate compound in the form of ethyl pyruvate in the preparation of an oral supplement for use in enhancing cell metabolism and absorption of medications in a human or animal body. According to a further aspect of the invention there is provided the use of a pyruvate compound in the form of ethyl pyruvate in the preparation of topical preparation for use in ocular, mucous membrane, otic or transdermal application in a human or animal body.

The ethyl pyruvate may be used together with micro-clustered water, and in particular micro-clustered water produced according to the method as claimed in U.S. Patent No. 6,033,678.

The composition may include one or more stimulants selected from a group including sugars, taurine, caffeine or ginseng.

The composition aiso may include at least one preservative selected from a group including benzoic acid, sorbic acid or the parabens.

The composition also may include carbohydrates selected from ribose or dihydroxyacetone.

The composition also may include herbs, such as Garcinia Cambogia, or herbal extracts, such as (-) hydroxycitric acid.

The composition also may include essential oils, which may be any volatile plant oils having a flavour or odour of the plant from which it is extracted, and includes one or more components of that oil that contribute to the odour or flavour of that plant.

Moreover, the composition may include one or more of flavourings, colorants, vitamins, minerals, electrolytes, carbonated gas, amino acids, superoxide dismutase, nutritive and/or non-nutritive sweeteners, peptides, proteins, omega fatty acids, fulvic acid, Coenzyme Q10 and/or aloe to facilitate ingestion and increase taste. DETAILED DESCRIPTION OF THE INVENTION

Example 1

A group of 12 athletes (19 - 22 years of age) self-administered oral ethyl pyruvate in solution (about 2 litres per day of a solution of about 0.06 weight-percent ethyl pyruvate in water) during daily workouts (approximately 2 hours duration) for four days. The athletes, having ingested oral ethyl pyruvate, reported less fatigue at the end of each workout, and similar favourable reports continued each day for the four days of practice. Each subject additionally reported better performance in high-activity training regimens (e.g., "wind sprints" as known in the art) relative to performance reported in practice sessions prior to oral ingestion of ethyl pyruvate.

Example 2

A sample of 10 healthy, less than 45 years old, highly physically and mentally fit male and female athletes were recruited from local military, cycling and tri-athlete populations to participate in a 14-day treatment study. These individuals were randomly assigned to one of two groups: (i) one group was the control group, while (ii) the other group consumed terahertz treated water that included ethyl pyruvate ("CPR®").

Each subject performed a baseline maximal exercise test. The exercise test was continued to the limit of tolerance on a cycle ergo meter with a work rate increased in increments of 20 Watts/min (Wasserman). Gas exchange measurements were taken continuously using a Vista Mini CPX O₂ and CO₂ Analyzer (Vacuumed) to estimate anaerobic or ventilation threshold and maximal aerobic power. The rating of perceived exertion, the respiratory exchange ratio and the time taken to complete the VO₂ max test were also determined. Additionally, heart rate response was analyzed at minutes 1, 2 and 5 and O₂ uptake was measured continuously for 15 minutes during recovery. Baseline Testing: No stimulants were taken on the day of testing. At sixty (60) minutes prior to testing, each subject was instructed either to consume 0.5 litre of CPR® {test group) or 0.5 litre of standard bottled water (placebo). Thirty (30) minutes prior to testing the subjects again consumed either 0.5 litre of CPR® or 0.5 litre placebo.

Water consumption after baseline testing: After the completion of the baseline testing, the treatment group began to consume terahertz treated water. The control group continued to consume placebo. The total period of consumption was 14 days. All water (treated or standard) was consumed between 5AM and 5PM each day of the evaluation. The test group subjects were not given terahertz treated water after 5PM, but were allowed to consume standard water during the evening. There were no other dietary or activity restrictions.

At the conclusion of the 14-day treatment period, all subjects underwent exercise testing identical to the baseline measurements.

The results are set out in Tables 1 and 2 and corresponding graphs 1 and 2. Table 1 details the V0₂ml/kg for each test subject, which is the measure of extraction and utilization of oxygen during the stationary bicycle test of these elite athletes. Table 2 details the power or maximum watts generated during testing.

Both sets of measurements were obtained at the anaerobic or ventilation threshold (VT), which is the transition point where the body must begin to utilize anaerobic (non-oxygenated) biochemical energy, having exhausted the aerobic (oxygen dependant) energy. Essentially these measurements were taken after the athletes "hit the wall" physiologically.

Tables 1 and 2 are divided in half and represent the control (placebo) and test (CPR®) consuming groups respectively. Subjects (SID) 1 , 3, 5, 7 and 9 were the control or placebo participants, whereas SID 2, 4, 6, 8 and 12 comprised the test group. The terms "condif and "CPRdif represent the calculated difference in repeat measurements (Test 1 and Test 2) under the same conditions for that subject. A mean value and percent change for each pair is listed.

Graphs 1 and 2, corresponding to Tables 1 and 2 respectively, reflect the mean % changes of either the control (left of midline) or CPR® (right of midline) group. These mean percent changes are listed vertically last under the column "change" and extreme right along the "mean" row. Hence the mean change in the power (Watts) table is 0.58% (contra!) versus 4.93% (placebo). The confidence limits "p" values are listed on the right hand side of each panel below the boxed grid and, for example, is listed below the V0₂ml/kg chart as 0.0027 (thus p < 0.01).

Conclusion

The following conclusions can be drawn from the above results, namely - (i) The pre-exercise consumption of CPR® during the 14-day treatment period had a measurable effect on the oxygen uptake, carbon dioxide excretion and ventilatory threshold as determined during pre- and post-treatment, graded maximal exercise testing; (ii) There was a significant reduction in the time it takes to complete a graded exercise test with CPR® as compared to standard water

(placebo) consumption prior to exercise; and (iii) The pre-exercise consumption of CPR® during the 14-day treatment period resulted in a significant decrease in the initial rate of recovery after maximal exercise testing.

Example 3

Eye lotion: Terahertz treated water, 0.9% sodium chloride, borate buffer system, 0.1% sorbic acid, up to 5% ethyl pyruvate, and 0.025% dipotassium EDTA. Example 4

Topical moisturising lotion: cyclomethicone, cyclomethicone-dimethicone, squalene, tocopherol acetate, grape seed oil, ethyl pyruvate/ester (up to 6%), jojoba oil, and fragrance.

Discussion

In human or animal bodies, pyruvate converts into acetyl-CoA through a process called "pyruvate dehydrogenase reaction". Acetyl-CoA is central to the balance between carbohydrate metabolism and fat metabolism. Normally, acetyl-CoA from fatty acid metabolism feeds into the citric acid cycle, contributing to the cell's energy supply. In the liver, when levels of circulating fatty acids are high, the production of acetyl-CoA from fat breakdown exceeds the cellular energy requirements. To make use of the energy available from the excess acetyl-CoA, ketone bodies are produced which can then circulate in the blood. In some circumstances this can lead to an excess of ketone bodies in the blood, a condition known as "ketoacidosis". This can occur in diabetes, starvation or in people following low-carbohydrate diets, all of which can cause fats to be metabolised as a major source of energy.

in plants, de novo fatty acid synthesis occurs in piastids. Many seeds accumulate large reservoirs of seed oils to support germination and early growth of the seedling before it is a net photosynthetic organism. Fatty acids are incorporated into membrane lipids, which is the major component of most membranes.

Acetoacetyl-CoA is the precursor to HMG-CoA, which is a vital component in cholesterol and ketone synthesis in human or animal bodys. Furthermore, it contributes an acetyl group to choline to produce acetylcholine, in a reaction catalysed by choline acetyltransferase.

In plants and animals, cytosolic acetyl-CoA is synthesized by ATP citrate lyase. When glucose is abundant in the blood of human or animal bodies, it is converted via glycolysis in the cytosol to pyruvate, and hence to acetyl- CoA in the mitochondrion. The excess of acetyl-CoA results in production of excess citrate, which is exported into the cytosol to give rise to cytosolic acety!-CoA.

Acetyl-CoA can be carboxyfated in the cytoso! by acetyl-CoA carboxylase, giving rise to maionyl-CoA, a substrate required for synthesis of flavonones and related polyketides, for elongation of fatty acids to produce waxes, cuticle, seed oils in members of the Brassica family, and for malonation of proteins and other phytochemicals.

Two acetyl-CoA can be condensed to create acetoacetyl-CoA, the first step in the HMG-CoA/ mevalonic acid pathway leading to synthesis of isoprenoids. !n plants these include sesquiterpenes, brassinosteroids (hormones) and membrane sterols.

The pyruvate dehydrogenase reaction is catalyzed by the pyruvate dehydrogenase complex. The enzyme consists of 60 subunits: 24 pyruvate dehydrogenase, 24 dihydrolipoyl transacetylase, and 12 dihydroiipoyl dehydrogenase (commonly denoted E1 , E2 and E3 respectively). The E1 subunits use TPP (thiamin pyrophosphate), the E2 subunits use lipoate and coenzyme A, and the E3 subunits use FAD and NAD⁺ as coenzymes.

The reaction of this complex follows three steps: Initially, pyruvate is bound by pyruvate dehydrogenase (E1) subunits and attacked at C2 by the zwitterionic form (negative charge about C2 of the thiazolium ring) of thiamin pyrophosphate, also bound by the enzyme. This tetrahedral intermediate undergoes decarboxylation resulting in an acyl anion equivalent (see cyanohydrin or aldehyde-dithiane umpolung chemistry, as well as benzoin condensation). This anion attacks the S1 of the oxidized lipoate species in an S_N2-like mechanism that displaces the S2 thiol as a sulfide or sulfhydryl moiety. Subsequent breakdown of the thiazole- hemithioacetal species ejects the TPP cofactor and generates an S1 thioester about the lipoate moiety. At this point, the lipoate-thioester functionality is translocated into the iipoate transacetylase (E2) active site, where it undergoes a transacylation with coenzyme A, generating dihydrolipoate and acetyl-CoA which subsequently enters the citric acid cycle.

The dihydroJipoate moiety then migrates to the dihydrolipoyl dehydrogenase (E3) active site where it undergoes FAD-mediated oxidation (identical in chemistry to disulfide isomerase) which returns lipoate to its resting state and generates FADH₂, which is further oxidized by the bound nicotinamide cofactor producing NADH₂ and regenerated flavin.

Ethyl pyruvate in particular, functions as an antioxidant. Oxidative stress occurs as a result from the production of reactive oxygen species, namely, hydrogen peroxide, hydroxyl radicals and superoxide radicals, among others, which are formed from both extracellular and intracellular sources. Reactive oxygen species are highly reactive species that, under normal conditions, are scavenged by endogenous defence mechanisms. However, under conditions of severe oxidative stress, reactive oxygen species interact with a variety of cellular components, causing peroxidation of lipids, denaturation of proteins, and interstitial matrix damage and resulting in increase of membrane permeability and release of tissue enzymes.

The applicants have found that orally administered ethyl pyruvate allows for the same cellular and organ benefits of intravenous ethyl pyruvate. No substantive change of the ethyl pyruvate molecule is necessary for effective cellular uptake. Also, the use of orally administered ethyl pyruvate compliment and supplement the actions of intravenous ethyl pyruvate for cardiac, muscular, nerve and other tissue resuscitation, and organ function enhancement in healthy and infirmed consumers not fully dependent upon intravenous ethyl pyruvate.

The ethyl pyruvate also functions to promote enhanced cellular function of all cells dependant on mitochondrial generated ATP for optimal function, gnome (DNA) self repair and normal cell life. The function of ethyl pyruvate further extends to promote increased overall energy, enhanced muscie strength endurance, decreased fatigue, increase mental alertness, and promote mood enhancement and a sense of well being.

During trials, the applicants have found that the use of orally administered ethyl pyruvate in combination with micro-clustered water, and particularly micro-clustered water produced according to method as claimed in U.S. Patent No. 6,033,678, produced exceptional results in test subjects. The combined use of the micro-clustered water with the oral ethyl pyruvate reduced the dosage requirements of the ethyl pyruvate with between 60% and 75%. Moreover, amateur athletes reported that the combined use of oral ethyl pyruvate and micro-clustered water significantly improved their strength, endurance and recovery times.

The ethyl pyruvate is also intended to increase cellular metabolism, fat metabolism, fat loss and enhance the actions of other weight loss products. When used specifically for weight loss, water and flavouring may be added to ethyl pyruvate to produce a liquid weight loss supplement.

Other functions of the ethyl pyruvate include promoting the effectiveness of protein powder, and decreasing muscle degradation in HIV/Aids and other muscle wasting and catabolic conditions.

The ethyl pyruvate according to the present invention also helps with decreasing blood glucose levels in diabetics, inhibiting the production of cell and tissue damaging free radicals and can further aid as an effective antioxidant, helping to eliminate formed free radicals. The purpose of the ethyl pyruvate may extend to contributing to the antioxidant activity of food preservatives or serving as an antioxidant itself.

Ethyl pyruvate is believed to improve cognitive, language, behavioural and social skills of autistic children and adults, promotes enhanced mental and physical recovery of cardiac, cancer and other chronically ill children and adults, and promotes enhanced energy recovery from mononucleosis, Chronic Fatigue Syndrome, Epstein Barr, hepatitis viruses and other chronic debilitating conditions.

Finally, this form of oral ethyl pyruvate allows for smaller quantities (by weight) of active pyruvate to be consumed compared to other oral pyruvate compositions for a desired physiologic effect.

Most, if not all of the above benefits are as a result of the ethyl pyruvate facilitating to increase membrane transfer and thus improve cellular absorption/delivery. Alternatively, without being bound by any particular scientific theory, ethyl pyruvate may interact with a specific membrane bound moiety or receptor (e.g. Sirtuin or PPAR gamma) signalling metabolic cascade(s) which end in observed energy dependant cellular response(s).

Tabie 1

Graph 1

Table 2

Graph 2

AVERAGE WATT (POWER) CHANGE AFTER 2 WEEKS OF USE (P<0 05)

■average wattchange

control CPR

Claims

1. A composition comprising pyruvate compound in the form of ethyl pyruvate and micro-clustered water.

2. The composition of claim 1 wherein the micro-clustered water is micro-clustered water produced according to the method as claimed in U.S. Patent No. 6,033,678.

3. The composition of claim 1 wherein the ethyl pyruvate is in liquid or powder form.

4. The composition of claim 1 wherein the composition is in the form of an oral supplement.

5. The composition of claim 4 wherein the oral supplement is included in a consumable beverage or food stuff.

6. The composition of claim 5 wherein the consumable beverage or foodstuff is an energy bar or diet bar, energy drink, soft drink, fruit juice, coffee- or tea-based drinks.

7. The composition of claim 6 wherein the tea-based drinks include Camellia sinensis var. sinensis and Camellia sinensis var. assamica, and include a blend of these teas.

8. The composition of claim 5 wherein the ethyl pyruvate is imprinted or infused with far infrared or other electromagnetic radiation before being incorporated into a consumable beverage or food stuff.

9. The composition of claim 4 wherein the oral supplement is introduced into a human or animal body in the form of a tablet or capsule.

10. The composition of claim 1 wherein the composition is in the form of a topical preparation.

11. The composition of claim 10 wherein the topical preparation is an ocufar, mucous membrane, otic or transdermal preparation.

12. The composition of claim 1 wherein the composition includes one or more stimulants selected from a group including sugars, taurine, caffeine and ginseng.

13. The composition of claim 1 wherein the composition includes at least one preservative selected from a group including benzoic acid, sorbic acid and the parabens.

14. The composition of claim 1 wherein the composition includes a carbohydrate selected from a group including ribose and dihydroxyacetone.

15. The composition of claim 1 wherein the composition includes a herb or herbal extract.

16. The composition of claim 15 wherein the herb is Garcinia Cambogia and the herbal extract is (-) hydroxycitric acid.

17. The composition of claim 1 wherein the composition includes an essential oil.

18. The composition of claim 17 wherein the essential oil is a volatile plant oil having a flavour or odour of the plant from which it is extracted, and includes one or more components of that oil that contribute to the odour or flavour of that plant.

19. The composition of claim 1 wherein the composition includes one or more components selected from a group including flavourings, colorants, vitamins, minerals, electrolytes, carbonated gas, amino acids, superoxide dismutase, nutritive and/or non-nutritive sweeteners, peptides, proteins, omega fatty acids, fulvic acid, Coenzyme Q10 and aloe to facilitate ingestion and increase taste.

20. A pyruvate compound in the form of ethyl pyruvate for use in enhancing cell metabolism and absorption of medications in a human or animal body.

21. The compound of claim 20 wherein the ethyl pyruvate is used together with micro-clustered water produced according to the method as claimed in U.S. Patent No. 6,033,678,

22. Use of a pyruvate compound in the form of ethyl pyruvate in the preparation of an oral supplement for use in enhancing cell metabolism and absorption of medications in a human or animal body.

23. Use of a pyruvate compound in the form of ethyl pyruvate in the preparation of topical preparation for use in ocular, mucous membrane, otic or transdermal application in a human or animal body.

24. The use of claims 22 or 23 wherein the ethyl pyruvate is used together with micro-clustered water produced according to the method as claimed in U.S. Patent No. 6,033,678.

25. A method of treating a human or animal body by administering a pyruvate compound in the form of ethyi pyruvate together with micro-clustered water produced according to the method as claimed in U.S. Patent No. 6,033,678 to the human or animal body to enhance cell metabolism and absorption of medications in the human or anima! body.

26. The composition of claim 1 substantially as herein described and exemplified.

27. The compound of claim 20 substantially as herein described and exemplified.

28. The use of claims 22 or 23 substantially as herein described and exemplified.