Application of ergothioneine in increasing creatine uptake

Abstract

The present invention discloses a method for enhancing the uptake of creatine in a tissue, comprising the use of ergothioneine and / or an ergothioneine derivative when creatine and / or a creatine derivative is used. The combined use of ergothioneine and creatine is disclosed to enhance athletic performance and / or cognitive function.

Description

Technical Field

[0001] This invention relates to the application of ergothioneine in increasing creatine uptake. Background Technology

[0002] Creatine's primary function in the body is to enhance energy storage by binding with phosphate groups to form phosphocreatine. Phosphocreatine, as an easily mobilized high-energy phosphate reserve, is crucial for the rapid regeneration of ATP during high-intensity exercise. Numerous studies have confirmed the efficacy of creatine; it provides explosive energy, enhances muscle strength, thereby improving athletic performance and results. Creatine also increases lean muscle mass and helps muscles recover faster during exercise, making it popular among athletes and fitness enthusiasts. In addition, creatine is present in the brain, providing energy to neurons. Scientists even believe that creatine is a neurotransmitter that supports brain health. Studies have shown that creatine supplementation may contribute to brain health and cognitive function (including memory, learning ability, concentration, reaction time, alertness, etc.), especially with age. Furthermore, some studies suggest that creatine may be beneficial for heart health because it can lower triglyceride and homocysteine ​​levels in the blood, substances associated with heart attacks and strokes. Creatine can also promote the production of more collagen and other substances in the skin, potentially helping to slow down skin aging.

[0003] Creatine enters the bloodstream after being synthesized by the body or supplemented exogenously, providing nutrition to all somatic cells. However, creatine cannot directly cross the cell membrane and requires a specific transport protein (creatine transporter, CrT) to assist it in crossing the membrane. In humans, high expression of creatine transporter is mainly found in organs that require creatine, such as the brain and skeletal muscle. Defects in creatine transporter can lead to severe muscle dysfunction, such as muscle weakness and atrophy, and can also harm brain health and impair cognitive function, as creatine cannot accumulate effectively in these tissues. Conversely, the activity and level of creatine transporter significantly affect tissue uptake of creatine. Studies have shown that the activity of creatine transporter may be nearing saturation; therefore, altering the level of creatine transporter may be a major determinant of intracellular creatine uptake. Increasing the expression level of creatine transporter would be an effective way to enhance creatine uptake by tissue cells, thereby increasing creatine and phosphocreatine content in tissues and further enhancing the efficacy of creatine.

[0004] Currently, commercially available creatine products include various forms of creatine, such as creatine monohydrate, creatine sulfate, and other creatine derivatives. These products are developed based on different mechanisms to enhance the effects of creatine, such as increasing the water content of muscle cells or increasing creatine absorption. Further enhancing the effects of creatine is a pressing technical challenge that needs to be addressed. Summary of the Invention

[0005] The purpose of this invention is to address the shortcomings of the prior art by providing a method for enhancing the uptake of creatine in tissues, comprising using creatine and / or creatine derivatives while simultaneously using ergothioneine and / or ergothioneine derivatives.

[0006] Furthermore, the use includes one or more of the following methods: oral administration, injection, sublingual absorption, and topical application; preferably oral administration.

[0007] Furthermore, it enhances the uptake of creatine in tissues, including the uptake of creatine by muscle tissue and / or the uptake of creatine by brain tissue. Furthermore, enhanced creatine uptake in tissues manifests as one or more of the following combinations: increased creatine accumulation, increased phosphocreatine content / reserves, increased cellular energy supply, increased muscle vitality, and increased brain energy.

[0008] Furthermore, the dosage of ergothioneine and / or ergothioneine derivatives is 5 mg-1000 mg per day, and the dosage of creatine and / or creatine derivatives is 500 mg-10 g per day; or the dosage of ergothioneine derivatives is the molar dosage of ergothioneine at 5 mg-1000 mg per day; or the dosage of creatine derivatives is the molar dosage of creatine at 500 mg-10 g per day.

[0009] Preferably, the dosage of ergothioneine and / or ergothioneine derivatives is 5 mg-500 mg per day, and the dosage of creatine and / or creatine derivatives is 500 mg-10 g per day; or the dosage of ergothioneine derivatives is the same as the dosage of ergothioneine calculated by molarity, which is 5 mg-500 mg per day; or the dosage of creatine derivatives is the same as the dosage of creatine calculated by molarity, which is 500 mg-10 g per day.

[0010] More preferably, the dosage of ergothioneine and / or ergothioneine derivatives is 10mg-500mg per day, and the dosage of creatine and / or creatine derivatives is 1g-10g per day; or the dosage of ergothioneine derivatives is the same as the dosage of ergothioneine calculated by molarity, which is 10mg-500mg per day; or the dosage of creatine derivatives is the same as the dosage of creatine calculated by molarity, which is 1g-10g per day.

[0011] Furthermore, ergothioneine and / or ergothioneine derivatives, including salts, esters, acids, ketones, polymers, cocrystals, chelates, complexes, glycosides, hydrates, and substances formed therefrom by non-chemical bonding with other substances. and / or Creatine and / or creatine derivatives, including salts, esters, ketones, hydrates, polymers, cocrystals, chelates, complexes, glycosides, and substances formed therefrom by non-chemical bonding with other substances.

[0012] This invention also provides the use of ergothioneine and / or ergothioneine derivatives in enhancing creatine uptake in tissues.

[0013] Furthermore, applications include the use of ergothioneine and / or ergothioneine derivatives when using creatine and / or creatine derivatives.

[0014] The present invention also provides a method for increasing the expression level of creatine transporter protein, characterized in that it includes using creatine and / or creatine derivatives while using ergothioneine and / or ergothioneine derivatives.

[0015] The use includes one or more of the following methods: oral administration, injection, sublingual absorption, and topical application; preferably oral administration.

[0016] Furthermore, it enhances the uptake of creatine in tissues, including the uptake of creatine by muscle tissue and / or the uptake of creatine by brain tissue. Furthermore, enhanced creatine uptake in tissues manifests as one or more of the following combinations: increased creatine accumulation, increased phosphocreatine content / reserves, increased cellular energy supply, increased muscle vitality, and increased brain energy.

[0017] Preferably, the dosage of ergothioneine and / or ergothioneine derivatives is 5 mg-500 mg per day, and the dosage of creatine and / or creatine derivatives is 500 mg-10 g per day; or the dosage of ergothioneine derivatives is the same as the dosage of ergothioneine calculated by molarity, which is 5 mg-500 mg per day; or the dosage of creatine derivatives is the same as the dosage of creatine calculated by molarity, which is 500 mg-10 g per day.

[0018] More preferably, the dosage of ergothioneine and / or ergothioneine derivatives is 10mg-500mg per day, and the dosage of creatine and / or creatine derivatives is 1g-10g per day; or the dosage of ergothioneine derivatives is the same as the dosage of ergothioneine calculated by molarity, which is 10mg-500mg per day; or the dosage of creatine derivatives is the same as the dosage of creatine calculated by molarity, which is 1g-10g per day.

[0019] Furthermore, ergothioneine and / or ergothioneine derivatives, including salts, esters, acids, ketones, polymers, cocrystals, chelates, complexes, glycosides, hydrates, and substances formed therefrom by non-chemical bonding with other substances. and / or Creatine and / or creatine derivatives, including salts, esters, ketones, hydrates, polymers, cocrystals, chelates, complexes, glycosides, and substances formed therefrom by non-chemical bonding with other substances.

[0020] The present invention also provides a composition comprising creatine and / or creatine derivatives, and ergothioneine and / or ergothioneine derivatives.

[0021] Ergothioneine and / or ergothioneine derivatives, including salts, esters, acids, ketones, polymers, cocrystals, chelates, complexes, glycosides, hydrates, and substances formed therefrom by non-chemical bonding with other substances. and / or Creatine and / or creatine derivatives, including salts, esters, ketones, hydrates, polymers, cocrystals, chelates, complexes, glycosides, and substances formed therefrom by non-chemical bonding with other substances.

[0022] Preferably, ergothioneine and / or ergothioneine derivatives are in a ratio of 0.0005:1 – 2:1, 0.0005:1 – 1:1, or 0.001:1 – 0.5:1 to creatine derivatives.

[0023] This invention also provides a combination of ergothioneine and creatine to enhance athletic performance and / or cognitive function.

[0024] Furthermore, applications include the use of ergothioneine and / or ergothioneine derivatives when using creatine and / or creatine derivatives.

[0025] Preferably, the dosage of ergothioneine and / or ergothioneine derivatives is 5 mg-500 mg per day, and the dosage of creatine and / or creatine derivatives is 500 mg-10 g per day; or the dosage of ergothioneine derivatives is the same as the dosage of ergothioneine calculated by molarity, which is 5 mg-500 mg per day; or the dosage of creatine derivatives is the same as the dosage of creatine calculated by molarity, which is 500 mg-10 g per day.

[0026] More preferably, the dosage of ergothioneine and / or ergothioneine derivatives is 10mg-500mg per day, and the dosage of creatine and / or creatine derivatives is 1g-10g per day; or the dosage of ergothioneine derivatives is the same as the dosage of ergothioneine calculated by molarity, which is 10mg-500mg per day; or the dosage of creatine derivatives is the same as the dosage of creatine calculated by molarity, which is 1g-10g per day.

[0027] Furthermore, ergothioneine and / or ergothioneine derivatives, including salts, esters, acids, ketones, polymers, cocrystals, chelates, complexes, glycosides, hydrates, and substances formed therefrom by non-chemical bonding with other substances. and / or Creatine and / or creatine derivatives, including salts, esters, ketones, hydrates, polymers, cocrystals, chelates, complexes, glycosides, and substances formed therefrom by non-chemical bonding with other substances.

[0028] Furthermore, it enhances athletic performance and / or cognitive function by one or more of the following: increased creatine accumulation, increased phosphocreatine content / reserves, increased cellular energy supply, increased muscle vitality, and increased brain energy.

[0029] Furthermore, enhancing athletic performance includes: increasing one or more of the following: muscle strength, muscle endurance, muscle power, and muscle recovery; and / or cognitive function includes: one or more of the following: memory, learning ability, concentration, reaction time, and alertness. Detailed Implementation

[0030] To facilitate understanding by those skilled in the art, the present invention will be further described below with reference to embodiments. The content mentioned in the embodiments is not intended to limit the present invention. Example 1

[0031] A method for enhancing the uptake of creatine in tissues includes using creatine and / or creatine derivatives while simultaneously using ergothioneine and / or ergothioneine derivatives.

[0032] The medication is administered orally.

[0033] The dosage of ergothioneine and / or ergothioneine derivatives is 5 mg per day, and the dosage of creatine and / or creatine derivatives is 500 mg per day. Example 2

[0034] A method for enhancing the uptake of creatine in tissues includes using creatine and / or creatine derivatives while simultaneously using ergothioneine and / or ergothioneine derivatives.

[0035] The medication is administered orally.

[0036] The dosage of ergothioneine and / or ergothioneine derivatives is 1000 mg per day, and the dosage of creatine and / or creatine derivatives is 10 g per day. Example 3

[0037] A method for enhancing the uptake of creatine in tissues includes using creatine and / or creatine derivatives while simultaneously using ergothioneine and / or ergothioneine derivatives.

[0038] The medication is administered orally.

[0039] The dosage of ergothioneine and / or ergothioneine derivatives is 100 mg per day, and the dosage of creatine and / or creatine derivatives is 5 g per day. Example 4

[0040] A method for enhancing the uptake of creatine in tissues includes using creatine and / or creatine derivatives while simultaneously using ergothioneine and / or ergothioneine derivatives.

[0041] The use is by injection.

[0042] The dosage of ergothioneine and / or ergothioneine derivatives is 50 mg per day, and the dosage of creatine and / or creatine derivatives is 3 g per day.

[0043] Experimental Example 1: Ergothioneine increases the expression of creatine transporter protein. Cellular experiments: Mouse myoblasts were cultured at 37°C in 5% CO2 high-glucose DMEM medium supplemented with 10% fetal bovine serum, penicillin, and streptomycin. The medium was changed every 48 hours. Cells were cultured at a density of 2.5 × 10³ cells per well in flat-bottomed 96-well plates, with different doses (0–10 mM) of ergothioneine added for 24 hours. After washing, cells were lysed with cell lysis buffer and then centrifuged at 13,000 rpm for 15 minutes at 4°C to precipitate cell debris. Protein levels were then detected by immunohistochemistry.

[0044] Table 1. Effects of different concentrations of ergothionein on the expression levels of creatine transporter proteins.

[0045] Note: Values ​​are presented as mean ± SEM. As shown in Table 1, ergothioneine increases the expression level of creatine transporter in a dose-dependent manner.

[0046] Experimental Example 2: Ergothioneine increases the uptake of creatine by skeletal muscle and brain tissue. Animal Experiment 1: SD rats were used and randomly divided into 7 groups after the first week of acclimatization. The groups were: control group, ergothioneine group (EGT, gavage dose 90 mg / kg / day, corresponding to a human dose of 1000 mg / day), and creatine monohydrate group (CR, gavage dose 900 mg / day, corresponding to a human dose of 10 g / day). Four groups were administered the following combinations: EC1 (0.45 mg / kg / day ergothioneine + 45 mg / kg / day creatine, equivalent to the human dose of 5 mg / day ergothioneine + 500 mg / day creatine), EC2 (4.5 mg / kg / day ergothioneine + 450 mg / kg / day creatine, equivalent to the human dose of 50 mg / day ergothioneine + 5 g / day creatine), EC3 (90 mg / kg / day ergothioneine + 900 mg / kg / day creatine, equivalent to the human dose of 1000 mg / day ergothioneine + 10 g / day creatine), and EC4 (0.45 mg / kg / day ergothioneine + 900 mg / kg / day creatine, equivalent to the human dose of 5 mg / day ergothioneine + 10 g / day creatine). The rats were fed for four months with free access to water and food. Thirty minutes after gavage on the last day, the rats were anesthetized and sacrificed. After dissection, brain and muscle tissues were cryopreserved in liquid nitrogen for subsequent chemical analysis. Brain and muscle tissues were homogenized in citrate buffer at a ratio of 1.3 g tissue / 7 mL buffer using an electric homogenizer. Then, 10 μg / mL internal standard solution was added to 100 μL of sample, vortexed, and acetonitrile was added. The mixture was centrifuged at 15,000 rpm for 5 minutes at 4 °C, and the supernatant was transferred to clean test tubes and dried using a rapid vacuum concentrator. The creatine concentration in brain and muscle tissues was detected by LC-MS / MS. The results are shown in the table below: Table 2 Creatine content in brain and muscle tissues of each group

[0047] Note: Values ​​are presented as mean ± SEM. The experimental results showed that, except for the EGT group rats supplemented with ergothioneine alone, whose brain and muscle tissue creatine content was not different from the control group, all other experimental groups showed varying degrees of increase, indicating that these groups all increased the uptake of creatine in the brain and muscle tissues of rats. Specifically, compared to the CR rats supplemented with creatine alone, the EC3 and EC4 groups, which were supplemented with high and low doses of ergothioneine (90 mg / kg / day and 0.45 mg / kg / day, respectively) to the same dose of creatine (900 mg / kg / day), both groups showed increased creatine content in brain and muscle tissues. In the EC3 group, the creatine content in both tissues was significantly increased compared to the CR group (p < 0.05), indicating that adding a high dose of ergothioneine can significantly increase the uptake of creatine in brain and muscle tissues. Although there was no significant difference in creatine content in both tissues in the EC4 group compared to the CR group, there was still a slight increase, indicating that a low dose of ergothioneine still increased the uptake of creatine in tissues, showing a dose-dependent trend.

[0048] Experiment 3: Combined use of ergothioneine and creatine enhances muscle strength in mice. Animal Experiment 2: KM mice were randomly divided into the following groups after one week of acclimatization: control group (con), ergothioneine group (EGT-m, 4.5 mg / kg / day), creatine group (CR-m, 450 mg / kg / day), and combination group (EC-m, 4.5 mg / kg / day ergothioneine + 450 mg / kg / day creatine). Mice were administered the food via gavage for 4 months, with free access to water and food. Muscle strength was measured 30 minutes after the last gavage. Muscle strength was measured by grip strength, using a grip strength meter to measure the grip strength of the mice's limbs. The results are shown in the table below: Table 3. Muscle strength of mice in each group

[0049] Note: Data is presented as mean ± SEM. The experimental results showed that, except for the EGT-m group (supplemented with ergothioneine alone), which did not show a significant increase in muscle strength compared to the control group (con), all other experimental groups showed a significant increase in muscle strength compared to the con group (p < 0.05). The EC-m group (combined with ergothioneine) showed a further significant increase in muscle strength compared to the CR-m group (supplemented with creatine alone) (p < 0.05). These results indicate that ergothioneine supplementation can significantly enhance the effect of creatine, confirming the previously verified conclusion that ergothioneine enhances creatine uptake in tissues. Increased tissue uptake of creatine means increased creatine accumulation in tissues, leading to increased creatine and phosphocreatine levels, thus further improving athletic performance. Studies have shown that ergothioneine can regulate the expression of various proteins and modulate cellular homeostasis. Furthermore, ergothioneine can accumulate intracellularly with a half-life of up to one month, meaning that even low doses of ergothioneine can be effective through long-term supplementation.

[0050] It will be understood by those skilled in the art that, unless otherwise defined, all terms used herein (including technical and scientific terms) have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. It should also be understood that terms such as those defined in general dictionaries should be understood to have the same meaning as in the context of the prior art, and should not be interpreted in an idealized or overly formal sense unless specifically defined as herein.

[0051] It should be understood that the above detailed description of the technical solutions of the present invention with reference to preferred embodiments is illustrative and not restrictive. Those skilled in the art can modify the technical solutions described in the embodiments or make equivalent substitutions for some of the technical features based on reading this specification; however, these modifications or substitutions do not cause the essence of the corresponding technical solutions to depart from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A method for enhancing the uptake of creatine in a tissue, characterized in that, The use of ergothioneine and / or ergothioneine derivatives is simultaneous with the use of creatine and / or creatine derivatives.

2. The method of claim 1, wherein, The use includes one or more than two combinations of oral, injection, sublingual absorption, topical external use; preferably oral.

3. The method of claim 1, wherein, The uptake of creatine in tissues, including the uptake of creatine by muscle tissue and / or the uptake of creatine by brain tissue, is enhanced; Further, the enhanced uptake of creatine in tissues is manifested as one or more than two combinations of increased creatine accumulation, increased phosphocreatine content / reserve, increased cell energy supply, increased muscle activity, and increased brain energy.

4. The method of claim 1, wherein, The dosage of ergothioneine and / or ergothioneine derivatives is 5mg-1000mg per day, and the dosage of creatine and / or creatine derivatives is 500mg-10g per day; Or the dosage of ergothioneine derivatives is 5mg-1000mg per day in terms of the dosage of ergothioneine in terms of substance amount; or the dosage of creatine derivatives is 500mg-10g per day in terms of the dosage of creatine in terms of substance amount; And / or Ergothioneine and / or ergothioneine derivatives, including salts, esters, acids, ketones, polymers, co-crystals, chelates, complexes, glycosides, hydrates of ergothioneine, and substances formed by non-chemical bonds with other substances; And / or Creatine and / or creatine derivatives, including salts, esters, ketones, hydrates, polymers, co-crystals, chelates, complexes, glycosides of creatine, and substances formed by non-chemical bonds with other substances.

5. A method for increasing the expression level of a creatine transporter, characterized by, The use of ergothioneine and / or ergothioneine derivatives is simultaneous with the use of creatine and / or creatine derivatives.

6. A composition characterized in that, The use of ergothioneine and / or ergothioneine derivatives is simultaneous with the use of creatine and / or creatine derivatives.

7. The composition of claim 6, wherein, Ergothioneine and / or ergothioneine derivatives, including salts, esters, acids, ketones, polymers, co-crystals, chelates, complexes, glycosides, hydrates of ergothioneine, and substances formed by non-chemical bonds with other substances; And / or Creatine and / or creatine derivatives, including salts, esters, ketones, hydrates, polymers, co-crystals, chelates, complexes, glycosides of creatine, and substances formed by non-chemical bonds with other substances.

8. The composition of claim 6, wherein, The ratio of ergothioneine and / or ergothioneine derivatives to creatine and / or creatine derivatives is 0.0005:1 – 2:1, preferably 0.0005:1 – 1:1, and further preferably 0.001:1 – 0.5:

1.

9. The use of ergothioneine and creatine in combination for enhancing athletic performance and / or cognitive function.

10. The use according to claim 9, characterized in that, The enhanced athletic performance includes one or more than two combinations of increased muscle strength, muscle endurance, muscle explosive force, and muscle recovery; And / or The cognitive function includes one or more than two combinations of memory, learning ability, concentration, reaction, and alertness.

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