TW201406381A - Pharmaceutical composition for anti-exercise fatigue - Google Patents

Abstract

A pharmaceutical composition for anti-exercise fatigue is provided. The pharmaceutical composition comprises a pharmaceutically acceptable carrier and an effective amount of an active component selected from at least one of the group consisting of a compound of formula (I), a pharmaceutically acceptable salt of the compound, and a pharmaceutically acceptable ester of the compound:

Description

Anti-sports fatigue pharmaceutical composition

The present invention relates to a pharmaceutical composition for anti-sports fatigue, and more particularly to an anti-exercise fatigue pharmaceutical composition comprising rutin.

Fatigue is a physiological message that usually refers to a feeling of tiredness, drowsiness, and lack of energy and motivation. According to the factors causing fatigue, fatigue can be roughly classified into two types: mental fatigue and physical fatigue. Possible factors that cause mental fatigue include emotional factors such as stress, depression, and sadness, while possible causes of physical fatigue include diseases (such as colds, anemia, and diabetes), lifestyle (smoking, drinking, and staying up late), and excessive Exercise and so on.

In general, fatigue usually disappears after the factors causing fatigue are removed and rested adequately. However, if the body has persistent or repeated episodes of intense fatigue for more than 6 months, it may have chronic fatigue syndrome (CFS). It is currently believed in medicine that chronic fatigue syndrome may be caused by multiple factors, including factors such as viral infection, immune system diseases, hepatitis, and chronic inflammation.

The fatigue caused by excessive exercise is called exercise fatigue. When the body is exercising, the muscles will consume nutrients such as oxygen, glucose, and electrolytes, and produce lactic acid. When oxygen and nutrients continue to be consumed and lactic acid continues to accumulate, the muscles will become stiff due to the shrinking function, and muscle fibers may become It causes tears due to severe contraction, which causes discomfort such as muscle soreness and muscle weakness. If you are tired of exercise Mildly, it can be recovered by proper physical rest such as rest, nutrient supplementation, massage and hot compress. However, if the fatigue is more serious, it should be treated with drugs such as muscle relaxants or non-steroidal anti-inflammatory drugs.

However, conventional drugs for treating sports fatigue are usually accompanied by different degrees of side effects. For example, muscle relaxants may cause side effects such as diarrhea, dry mouth, and lethargy; non-steroidal anti-inflammatory drugs may cause damage to the mucosa of the digestive tract, abnormal liver function, Side effects such as dizziness, headache, and lethargy. Therefore, there is still a need for a drug that can be used for anti-sports fatigue and has low side effects to effectively prevent and treat related symptoms caused by exercise fatigue.

The present invention is directed to the above-mentioned needs, and the inventors of the present invention found that rutin can effectively inhibit exercise fatigue, and thus can be used for preventing or treating related symptoms caused by exercise fatigue.

It is an object of the present invention to provide a pharmaceutical composition for anti-exercise fatigue comprising an effective amount of an active ingredient and a pharmaceutically acceptable carrier, the active ingredient being selected from at least one of the following groups: (I) Compounds and pharmaceutically acceptable salts and esters thereof:

Another object of the invention is the use of a compound of formula (I), a pharmaceutically acceptable salt thereof, and/or a pharmaceutically acceptable ester thereof, for the manufacture of a medicament, wherein the medicament is for use in the manufacture of a medicament Exercise fatigue.

The detailed description of the present invention and the preferred embodiments thereof will be described in the following description.

The invention will be described in detail below with reference to the specific embodiments of the present invention. The invention may be practiced in various different forms without departing from the spirit and scope of the invention. The person stated. In addition, the terms "a", "an" and "the"

As described above, when a living body moves, it causes muscle soreness, stiffness, weakness, and the like due to nutrient consumption and accumulation of lactic acid. The inventors of the present study have found that rutin can achieve anti-sports fatigue by delaying the occurrence of exercise fatigue and reducing the amount of blood lactic acid.

Accordingly, the present invention provides a pharmaceutical composition for anti-exercise fatigue comprising an effective amount of an active ingredient and a pharmaceutically acceptable carrier selected from at least one of the group consisting of: Compounds of formula (I) and pharmaceutically acceptable salts and esters thereof:

Preferably, the active ingredient is a compound of formula (I).

The compound of formula (I) is rutin, which is a biological flavonoid, a known organism. Flavonoids have the effect of strengthening microvasculature and can therefore be used to treat blood stasis, varicose veins, and acne. Rhodamine is widely found in a variety of vegetables and grains, and has anti-inflammatory, antibacterial, anti-cancer effects, and can slow the release of histamine from mast cells. Therefore, since histamine is a key chemical substance causing allergic reactions, Muscatin can have anti-allergic effects by reducing histamine.

Studies have shown that exercise fatigue is associated with the expression of genes and proteins of the peroxisome proliferator-initiating receptor γ-co-priming factor-1α (PGC-1α) and the silencing regulator T1 (SIRT1). It is known that PGC-1α and SIRT1 are important proteins that promote the mechanism of mitochondrial biogenesis of cells. When the expression of PGC-1α and SIRT1 protein in cells increases, it promotes the synthesis of mitochondria and makes cells The mitochondrial content is increased, thereby supplying more energy to the organism (adenosine triphosphate; ATP), which increases the exercise tolerance and anti-fatigue effect of the organism (see J. Mark Davis et al., Quercetin increases). Brain and muscle mitochondrial biogenesis and exercise tolerance. Am J Physiol Regul Integr Comp Physiol 296: R1071-R1077, 2009, the entire disclosure of which is hereby incorporated by reference. The pharmaceutical composition of the invention can promote the transcription and protein expression of the peroxisome proliferator-initiating receptor γ-co-priming factor-1α and the silencing information regulator T1, and has the effect of resisting exercise fatigue.

On the other hand, as mentioned above, the nutrients consumed during exercise are converted into lactic acid. If the produced lactic acid is not metabolized or excreted in time, it will stimulate the nerve endings in the muscle due to excessive accumulation of lactic acid, or cause osmotic pressure. The change causes the water to increase and penetrate into the muscle fibers, causing the muscles to swell, resulting in muscle soreness. It has been found that the pharmaceutical composition of the present invention can reduce the amount of lactic acid in the blood, thereby achieving the effect of suppressing exercise fatigue.

The pharmaceutical compositions of the present invention can be used in veterinary and human medicine, and can be administered in any form and in any convenient manner. For example, but not limited thereto, the pharmaceutical composition can be administered by oral, subcutaneous, intravenous or intra-articular injection. Depending on the form of use and the use, a pharmaceutically acceptable carrier can be included in the pharmaceutical compositions of the present invention.

For example, in the preparation of a pharmaceutical preparation suitable for oral administration, a pharmaceutical acceptable carrier which does not adversely affect the activity of the aromatin can be contained in the pharmaceutical composition of the present invention, for example, a solvent, an oily solvent, a diluent, a stabilizer, and a absorption delay. Agents, disintegrating agents, emulsifiers, antioxidants, binders, lubricants, moisture absorbents, and the like. The composition may be formulated into a form suitable for oral administration by any convenient method, for example, a tablet, a capsule, a granule, a powder, a flow extract, a solution, a syrup, a suspension, an emulsion, and Tincture and so on.

As for the pharmaceutical form suitable for subcutaneous, intravenous or intra-articular injection, one or more, for example, isotonic solutions, salt buffers (such as phosphate buffer or citrate buffer) may be included in the pharmaceutical composition of the invention. A solubilizing agent, an emulsifier, and other carriers are prepared, for example, as an intravenous infusion solution, an intravenous infusion of an emulsion, a dry powder injection, a suspension injection, or a dry powder suspension injection.

The pharmaceutical composition of the present invention may further contain additives such as a flavoring agent, a toner and a coloring agent as needed to improve the mouthfeel and visual feeling when the obtained medicament is taken; and a reasonable amount of preservative, preservative, antibacterial agent, An antifungal agent or the like to improve the storage property of the resulting agent.

Optionally, the pharmaceutical composition of the present invention may contain one or more other active ingredients to further enhance the efficacy of the pharmaceutical composition of the present invention or to increase the use of the formulation. Flexibility and deployment. For example, the pharmaceutical composition of the present invention may contain one or more of the following active ingredients: vitamins, whey proteins, muscle relaxants, non-steroidal anti-inflammatory drugs, and other active ingredients, as long as the other active ingredients are against the ruthenium There is no adverse effect on the benefits.

The invention also provides the use of a compound of formula (I), a pharmaceutically acceptable salt thereof, and/or a pharmaceutically acceptable ester thereof, for the manufacture of a medicament, wherein the medicament is for use in combat fatigue. Specifically, the agent can be used to promote gene transcription and protein expression of the peroxisome proliferator-initiating receptor γ-co-priming factor-1α and the sirtuin T1, and to reduce the amount of lactic acid in the blood. In addition, the agent can be administered at different dosing times, such as once a day, multiple times a day, or once a day, depending on the needs of the target. For example, when used to inhibit exercise fatigue, the amount of the agent is from about 10 mg/kg body weight to about 80 mg/kg body weight per day, preferably about 15 mg/kg body weight per day, based on the compound of formula (I). Up to about 70 mg / kg body weight, wherein the unit "mg / kg body weight" refers to the amount of drug required per kilogram of body weight.

The invention is further illustrated by the following specific embodiments. The embodiments are provided by way of illustration only and are not intended to limit the scope of the invention.

[Examples] [Example 1] Mouse weight swimming test and blood lactate amount determination

(1) rutin

The mouse weight-bearing swimming experiment mode was used to test whether the rutin was effective against exercise fatigue. Six-week-old healthy adult mice were selected, and the mice were divided into four groups. The initial body weight of each group of mice was measured before the administration of muscidin, and then administered by needle feeding. Mg/kg body weight (control group, only solvent (2.5% DMSO)), 15 mg/kg body weight, 30 mg/kg body weight, and 60 mg/kg body weight of rutin (purchased from Sigma Chemical, Missouri, US), once daily, after 7 days of continuous feeding, the final body weight of each group of mice was measured, and the results are shown in Table 1. (2) Weight-bearing swimming experiment

One week before the weight-bearing swimming experiment, after 30 minutes of feeding, swimming was first adapted. In the 30 to 60 minutes after the last feeding, the mice were tied with lead wire (loading 2% to 5% of the body weight of the mouse) and placed in a glass water having a diameter of 15 cm, a water depth of 20 cm, and a water temperature of 27 ± 1 °C. In the tank, the mice were forced to swim, and the swimming time of each group of mice was recorded until the body's physical exertion was exhausted, and it was not able to surface until the head watered for 8 seconds. The results are shown in Table 1. A related experimental method for a mouse weight-bearing swimming test can be found in Antifatigue Activity of Phenylethanoid-rich Extract from Cistanche Deserticola, PHYTOTHERAPY RESEARCH, 24: 313-315 (2010), which is incorporated herein by reference in its entirety.

The results in Table 1 show that the weight-bearing swimming time of the mice administered with rutin was prolonged, and the final weight of the control group mice not administered with rutin and the mice administered with rutin There was no significant difference, indicating that the prolongation of the weight-bearing swimming time in mice was caused by the administration of muskin, not by the difference in body weight of the mice. In addition, the higher the dose of rutin, the longer the swimming time of the mice. The results of this experiment show that rutin can inhibit exercise fatigue and prolong the exercise time of mice, so it has anti-fatigue effect.

(3) Lactic acid determination experiment

Before the above-mentioned weight-bearing swimming test was performed on the mice, the amount of blood lactate (initial blood lactate amount) was recorded; and after swimming for 10 minutes under weight-bearing, the amount of blood lactate in the mice (the amount of blood lactate after weight-bearing swimming) was recorded immediately, and then After the mice were rested for 20 minutes, the amount of blood lactic acid in the mice (final blood lactic acid amount) was measured again. The lactic acid assay experiment uses the principle of enzyme action and colorimetric assay to add lactate oxidase to the quantitative plasma, followed by 4-aminoantipyrine and 1,7-dihydroxyl. 1,7-dihydroxynaphthalene, after the action of hydrogen peroxide, produces a red compound, and its absorbance is measured at a wavelength of 540 nm, and then the concentration of lactic acid is converted. The average of the experimental results is shown in Table 2.

The lactic acid amount elimination ratios listed in Table 2 are calculated by the following formula: The greater the elimination ratio, the more significant the effect of reducing the amount of blood lactate after 20 minutes of rest after exercise. The results in Table 2 show that the elimination ratios of the experimental mice in Groups 1, 2, and 3 (15, 30, and 60 mg/kg body weight of the rutin doses, respectively) were larger than those of the control group administered with rutin. In rats, it has been shown that rutin can effectively reduce the amount of lactic acid in the blood in a short period of time after exercise, thereby achieving the effect of suppressing exercise fatigue.

[Example 2] Gene transcription test of PGC-1α and SIRT1

A polymerase chain reaction (PCR) test was performed to determine whether rutin could achieve anti-fatigue effects by affecting the degree of gene expression of PGC-1α and SIRT1. First, the mice were divided into three groups before the experiment, and the mice were administered with 0 mg/kg body weight by needle feeding (control group, only solvent (2.5% DMSO), 30 mg/kg body weight, and 60). The mg/kg body weight of rutin, once daily, after 7 days of continuous feeding, was carried out for 10 minutes after the weight-bearing swimming test. The mouse was sacrificed and the gastrocnemius muscle of the hind limb was removed. The surgical procedure was based on the Japanese Physiological Society. The Guiding Principles for the Care and Use of Animals in the Field of Physiological Sciences is conducted by the Society of Principles for the Care and Use of Animals in the Field of Physiological Sciences. Total RNA was then extracted with a TRI reagent (Molecular Research Center, Cincinnati, Ohio, USA). Quantitative polymerase chain reaction (Marligen, Biosciences Ijamsville, Maryland, USA) using the first-strand cDNA synthesis system to convert total DNA treated with DNase into cDNA, followed by PGC-1α and SIRT1 The primer of the gene was subjected to PCR (setting conditions: DNA denaturation at 94 ° C for 30 seconds; primer bonding at 55 ° C for 30 seconds; Extension at 94 ° C for 60 seconds, repeat 30 cycles, maintain at 72 ° C for 10 minutes, then cool to 4 ° C to end the reaction), the product obtained by PCR at 50 volts with 1.5% acacia The gel was subjected to electrophoresis analysis for 20 minutes, and then stained with EtBr dye for 10 minutes, and photographed to analyze the strength of the band on the colloid. The results are shown in Fig. 1, in which glyceraldehyde-3-phosphate dehydrogenase (glyceraldehyde-3) was used. -phosphate dehydrogenase, GADPH) as an internal control group.

The results in Fig. 1 show that the gene expression levels of PGC-1α and SIRT1 in the mice administered with rutin are significantly increased compared with those in the control group without the administration of rutin, indicating that rutin can promote PGC- The genes of 1α and SIRT1 are transcribed and have anti-sports fatigue effects.

[Example 3] Protein expression test of PGC-1α and SIRT1

The protein expression of PGC-1α and SIRT1 was further analyzed by Western blotting. First, the mice were divided into three groups before the experiment, and the mice were administered with 0 mg/kg body weight by needle feeding (control group, only solvent (2.5% DMSO), 30 mg/kg body weight, and 60). The mg/kg body weight of rutin, once daily for 7 days, after the weight-bearing swimming test for 10 minutes, sacrifice the mice and surgically remove the gastrocnemius muscle of the hind limbs of the mice, extract the total protein, and take 50 mg of the obtained protein. The electrophoresis was carried out at a voltage of 120 volts with a 1.5 mm thick discontinuous acrylamide gel (5% colloidal amide amide colloid and 12% acrylamide colloid). Next, the colloid was transferred to a PVDF membrane, and incubated in TBST buffer (24.22 g Tris, 87.75 g NaCl, 10 ml Tween 20, and water adjusted to 1 liter), and washed for 10 minutes 3 times, then the primary antibody was added. The reaction was shaken at 4 ° C for one day, and washed with TBST buffer for 10 minutes 3 times every other day. Then add secondary antibody, shake at room temperature for 1 hour, then shake with TBST buffer for 10 minutes 3 times, then After the PVDF membrane was reacted with enhanced chemi-luminescence (ECL), the cold light intensity was detected by a fully automatic luminescence analyzer. The results are shown in Fig. 2, where β-actin (ß-actin) was used. As an internal control group.

The results in Fig. 2 show that the protein expression levels of PGC-1α and SIRT1 in the mice administered with rutin are significantly increased compared with those in the control group without the administration of rutin, indicating that rutin can promote PGC- The protein of 1α and SIRT1 exhibits anti-sports fatigue effects.

The above embodiments are merely illustrative of the principles and effects of the invention and are not intended to limit the invention. Modifications and variations of the above-described embodiments can be made by those skilled in the art without departing from the spirit and scope of the invention. Therefore, the scope of protection of the present invention should be as set forth in the scope of the patent application described hereinafter.

Figure 1 is an electropherogram showing the transcription of PGC-1α and SIRT1 genes in mice, and Figure 2 is a Western blot analysis showing the protein expression of PGC-1α and SIRT1 in mice. Figure.

Claims (9)

A pharmaceutical composition for anti-sports fatigue comprising an effective amount of an active ingredient and a pharmaceutically acceptable carrier, the active ingredient being selected from at least one of the group consisting of a compound of formula (I) and a medicament thereof Acceptable salts and esters: The pharmaceutical composition according to claim 1, which is for promoting transcription and/or protein expression of a gene selected from at least one of the group consisting of a peroxisome proliferator-priming receptor γ-co-priming factor-1α (PGC-1α) And the silent information adjustment factor T1 (SIRT1). The pharmaceutical composition of claim 1, which is for reducing the amount of lactic acid in the blood. The pharmaceutical composition according to any one of claims 1 to 3, wherein the active ingredient is a compound of formula (I). Use of a compound of formula (I), a pharmaceutically acceptable salt thereof, and/or a pharmaceutically acceptable ester thereof for the manufacture of a medicament: The agent is used to combat exercise fatigue. The use of claim 5, wherein the amount of the agent is from about 10 to 80 mg/kg body weight per day based on the compound of formula (I). The use of claim 5, wherein the amount of the agent is from about 15 to 70 mg/kg body weight per day based on the compound of formula (I). The use of claim 5, wherein the agent is for promoting transcription and/or protein expression of a gene selected from at least one of the group consisting of peroxisome proliferator-priming receptor gamma co-priming factor-1 alpha (PGC-1α) And the silent information adjustment factor T1 (SIRT1). The use of claim 5, wherein the agent is for reducing the amount of lactic acid in the blood.