Application of alpha-ketobutyrate in preparation of medicine for preventing and / or treating skeletal muscle atrophy

Abstract

The invention provides application of alpha-ketobutyrate in preparation of drugs for prevention and / or treatment of skeletal muscle atrophy, and belongs to the technical field of biomedicine.The human endogenous metabolite alpha-ketobutyrate is applied to prevention and treatment of skeletal muscle atrophy, the side effect is small, the safety is high after long-term use, and the effect is obvious. Serious immunoreaction or toxic reaction can be avoided, and skeletal muscle senescence and muscular atrophy can be effectively prevented and treated. The alpha-ketobutyrate can improve the muscle index of skeletal muscles of the old mice, increase the number of muscle fibers, increase the cross sectional area, increase the number of muscle cell nucleuses and the expression of nuclear fiber layer protein LamB1 of the muscle cell nucleuses, and can also increase the capillary angiogenesis rate in muscle tissues of the old mice. Alpha-ketobutyrate has good pharmacological characteristics and low side effects, and provides an important basis for developing clinical drugs for treating amyotrophy.

Description

Technical Field

[0001] The present invention belongs to the technical field of biomedicine, and particularly relates to the application of α-ketobutyrate in the preparation of drugs for preventing and / or treating skeletal muscle atrophy. Background Art

[0002] Skeletal muscle aging refers to the degenerative changes in muscle mass, strength, and function with age, and muscle atrophy is an obvious sign of skeletal muscle aging. After the age of 50, ordinary people lose about 1% of muscle mass and 3% of muscle strength every year, and the cumulative net loss of muscle mass is at least 30% during the aging process. This loss may lead to inconvenience in movement for the elderly population and even loss of the ability to live independently. In addition, skeletal muscle atrophy is a key link in various chronic diseases of the elderly population (such as chronic obstructive pulmonary disease, heart failure, chronic kidney disease, cachexia, Duchenne muscular dystrophy, etc.), seriously threatening the health of the elderly and affecting the quality of life of the elderly. With the increasing aging of the world population, how to effectively prevent and treat skeletal muscle atrophy in the elderly population has become one of the important issues in the field of global public health.

[0003] Although a large number of studies have been carried out in recent years to clarify the pathogenesis of muscle atrophy, identify treatment targets, and promote drug development, and steady progress has been made, there are still many challenges in developing effective treatment methods and drugs for improving skeletal muscle atrophy. At present, the drug development for treating skeletal muscle atrophy mainly focuses on the following aspects: regulating skeletal muscle protein metabolism, such as Myostatin inhibitors; or metabolic regulators that improve energy metabolism and mitochondrial function, such as fibrates; or hormones that enhance muscle mass and strength, such as growth hormone, insulin-like growth factor-1, and testosterone, etc.; in addition, regenerative medicine centered on stem cell therapy promotes muscle regeneration by transplanting myoblasts or regulating satellite cell activation. The above treatment methods show certain effects in animal models, but the results in human trials are still not ideal. Hormone therapy is limited in clinical application due to its large side effects, and in addition, the related research on stem cell therapy is still in the preclinical or early trial stage. Summary of the Invention

[0004] In order to solve the problems of skeletal muscle aging and muscle atrophy, the present invention provides the application of α-ketobutyrate in the preparation of drugs for preventing and / or treating skeletal muscle atrophy. Applying the endogenous metabolite α-ketobutyrate in the human body to prevent and treat skeletal muscle atrophy has less side effects and higher safety for long-term use, can avoid triggering serious immune reactions or toxic reactions, and can effectively prevent and treat skeletal muscle aging and muscle atrophy.

[0005] The present invention is achieved through the following technical solutions:

[0006] The present invention provides the application of α-ketobutyrate in the preparation of drugs for preventing and / or treating skeletal muscle atrophy, and the structural formula of the α-ketobutyrate is:

[0007]

[0008] Based on the same inventive concept, the present invention provides the use of α-ketobutyrate in the preparation of a medicament for preventing and / or treating skeletal muscle aging.

[0009] Based on the same inventive concept, the present invention provides the use of α-ketobutyrate in the preparation of a medicament for improving the exhaustion performance of aged animals.

[0010] Based on the same inventive concept, the present invention provides the use of α-ketobutyrate in the preparation of a medicament for improving the skeletal muscle performance of aged mice, wherein the skeletal muscle performance of the aged mice includes the maintenance time of the aged mice on a rotarod.

[0011] Based on the same inventive concept, the present invention provides the use of α-ketobutyrate in the preparation of a medicament for improving the skeletal muscle physiological characteristics of aged animals, wherein the skeletal muscle physiological characteristics include at least one of the anterior tibial muscle index, the cross-sectional area of muscle fibers, the capillary formation rate in muscle tissue, the number of myocyte cell nuclei, and the expression of lamin B1 of the muscle cell nuclear lamina.

[0012] Based on the same inventive concept, the present invention provides a medicament for preventing and / or treating skeletal muscle atrophy, and the active ingredient of the medicament includes α-ketobutyrate.

[0013] Furthermore, the dosage form of the medicament includes any one of tablets, capsules, oral liquids, granules, effervescent tablets, and sustained-release agents.

[0014] Based on the same inventive concept, the present invention provides a medicament for preventing and / or treating skeletal muscle aging, and the active ingredient of the medicament includes α-ketobutyrate.

[0015] One or more technical solutions in the embodiments of the present invention have at least the following technical effects or advantages:

[0016] 1. Use of α-ketobutyrate in the preparation of a medicament for preventing and / or treating skeletal muscle atrophy. By intervening with α-ketobutyrate in aged mice, the present invention shows that it can significantly improve the exhaustion performance of aged mice and increase the maintenance time of mice on the rotarod. Further research finds that α-ketobutyrate can significantly improve the pathological phenotypes related to muscle atrophy in aged mice, including decreased muscle fiber number, reduced cross-sectional area, and decreased capillary angiogenesis rate, etc., improve the muscle index of skeletal muscle in aged mice, increase the number and cross-sectional area of muscle fibers, increase the number of muscle cell nuclei, and the expression of nuclear laminin LaminB1 in muscle cell nuclei, and can also increase the capillary angiogenesis rate in the muscle tissue of aged mice. In addition, as an endogenous metabolite, α-ketobutyrate has good pharmacological properties and low side effects, providing an important basis for the development of clinical drugs for treating muscle atrophy.

[0017] 2. Use of α-ketobutyrate in the preparation of a medicament for preventing and / or treating skeletal muscle atrophy. Since α-ketobutyric acid is an endogenous metabolite, compared with synthetic drugs, it has fewer side effects and higher safety during long-term use, and can avoid triggering serious immune reactions or toxic reactions. In addition, endogenous metabolites have good pharmacokinetic properties in the body and have multi-target effects, and can play a role by regulating multiple signaling pathways. This multiple targeting mechanism makes endogenous metabolites have great potential in the treatment of complex diseases. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] To more clearly illustrate the technical solutions in the embodiments of the present invention, the following will briefly introduce the drawings required for the description of the embodiments. Obviously, the drawings in the following description are some embodiments of the present invention. For those of ordinary skill in the art, without creative efforts, other drawings can also be obtained based on these drawings.

[0019] Figure 1 Effect of α-ketobutyrate on the maximum movement distance and longest movement time of aged mice.

[0020] Figure 2 Effect of α-ketobutyrate on the falling time of aged mice on the rotarod.

[0021] Figure 3 Effect of α-ketobutyrate on the muscle index of aged mice.

[0022] Figure 4 Double immunofluorescence staining of Laminin and Cd31 shows the effect of α-ketobutyrate on the morphology of aged muscle tissue.

[0023] Figure 5 Effect of α-ketobutyrate on the cross-sectional area of muscle fibers in aged mice.

[0024] Figure 6 Effect of α-ketobutyrate on capillary formation rate in muscle tissue of aged mice.

[0025] Figure 7 Effect of α-ketobutyrate on the number of myonuclei in muscle tissue of aged mice.

[0026] Figure 8 Immunofluorescence staining of lamin B1 to show the effect of α-ketobutyrate on muscle cell nuclei in aged muscle tissue. Detailed implementation manners

[0027] The present invention will be specifically described below in combination with specific implementation manners and examples, and the advantages and various effects of the present invention will be presented more clearly therefrom. Those skilled in the art should understand that these specific implementation manners and examples are used to illustrate the present invention, rather than limiting the present invention.

[0028] Throughout the specification, unless otherwise specifically stated, the terms used herein should be understood as having the meanings commonly used in the art. Therefore, unless otherwise defined, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art to which the present invention belongs. In case of contradiction, this specification shall prevail.

[0029] Unless otherwise specifically stated, various raw materials, reagents, instruments, and equipment used in the present invention can be obtained through market purchases or can be prepared by existing methods.

[0030] Unless otherwise specifically stated, the experimental methods used in the following examples are all conventional methods.

[0031] The application of α-ketobutyrate of the present invention in the preparation of drugs for preventing and / or treating skeletal muscle atrophy will be described in detail below in combination with examples and experimental data.

[0032] The molecular formula of α-ketobutyrate of the present invention is CH3CH2COCOONa, and the molecular weight is 124.07. The present invention has no special limitation on the source of α-ketobutyrate, and the sources of α-ketobutyrate well-known in the art can be used. In the examples of the present invention, the α-ketobutyrate was purchased from Shanghai Macklin Biochemical Co., Ltd., and the product number was S844402.

[0033] Example 1

[0034] Method for treating muscle atrophy in aged mice by administering α-ketobutyrate (α-KB).

[0035] 1. Method for raising mice

[0036] C57BL / 6J mice, 24 months old, half male and half female, SPF grade, weighing 28 ± 5 g, were purchased from the Experimental Animal Department of Kunming Medical University (SCXK(Dian)K2020-0004). Five mice were housed in each cage and kept in a 12-hour light-dark cycle and temperature-controlled environment. The temperature was 24 ± 2 °C, and the relative humidity was 50% ± 5%. Food and water were provided ad libitum. All animal experiments in this example were carried out in accordance with the regulations of animal experiments. The Animal Ethics Committee of Kunming Medical University reviewed and approved the animal ethics of this study.

[0037] 2. Administration dosage and method

[0038] Experimental group: α-ketobutyrate was dissolved in the drinking water of mice to a concentration of 0.2%, and the mice drank the water ad libitum. The administration time was 6 months.

[0039] Control group: Normal drinking water.

[0040] 3. Exhaustion test of mice

[0041] (1) Equipment used for the test: Electric roller running device (#YLS-10B, Yiyan Technology Development Co., Ltd., Jinan)

[0042] (2) Pre-adaptation: Before the formal test, 2-3 days of adaptive training was carried out. Mice ran on the treadmill every day, usually for 20-30 minutes, gradually adapting to the exercise intensity.

[0043] (3) Experimental parameter settings: The rotation speed was 13 rpm, and the electric shock current was 20 mA. After the mice were shocked, if they rested continuously 5 times within 10 minutes and each rest lasted for 30 seconds, it was regarded as exhaustion.

[0044] (4) Experimental results: Please refer to Figure 1 , treatment with 0.2% α-ketobutyrate could significantly improve the exhaustion performance of mice, mainly manifested as significant improvement in the maximum exercise distance and the longest exercise time.

[0045] 4. Rotarod test

[0046] (1) Equipment used for the test: Rotarod fatigue tester (#YLS-4D, Yiyan Technology Development Co., Ltd., Jinan)

[0047] (2) Pre-adaptation: Before the formal test, animals usually needed 1-2 days of adaptive training. During the adaptation period, mice were placed on the rotarod with a relatively low rotation speed to gradually adapt to the movement of the rotarod. Each training lasted about 5-10 minutes and was carried out 3 times a day.

[0048] (3) Experimental parameter settings: The rotation speed of the rotarod accelerated to 30 rpm within 5 minutes, and the time when the animals fell off the rotarod was observed and recorded.

[0049] (4) Experimental results: Please refer to Figure 2 , the use of 0.2% α-ketobutyrate treatment can significantly improve the maintenance time of mice on the rotarod.

[0050] 5. Detection of indicators related to skeletal muscle atrophy in mice

[0051] Weigh the body weight of the mice, sacrifice them after anesthesia, take the tibialis anterior muscle of the mice, weigh the above two muscles, fix them with 4% paraformaldehyde for 24 hours, dehydrate them with ethanol, clear them with xylene, embed them in paraffin, and section them. Use double immunofluorescence staining of Laminin and CD31 and LaminB1 immunofluorescence staining method to detect the indicators related to skeletal muscle atrophy.

[0052] Please refer to Figure 3 , the use of 0.2% α-ketobutyrate treatment can significantly increase the muscle index of mice.

[0053] Please refer to Figures 4-5 , the use of 0.2% α-ketobutyrate treatment can significantly increase the number and cross-sectional area of muscle fibers in the skeletal muscle of mice.

[0054] Please refer to Figure 4 ,6, the use of 0.2% α-ketobutyrate treatment can significantly increase the angiogenesis rate in the muscle tissue of old mice.

[0055] Please refer to Figure 4 ,7, the use of 0.2% α-ketobutyrate treatment can significantly increase the number of muscle nuclei.

[0056] Please refer to Figure 8 , the use of 0.2% α-ketobutyrate treatment can significantly increase the expression of laminin B1 in the nuclear lamina of muscle nuclei.

[0057] In summary, this invention uses C57BL / 6J naturally aged mice (24 months old) as a model, adds α-ketobutyrate to the drinking water of the mice to make its concentration 0.2%, the mice drink water freely, and the administration time is 6 months. The test results show that α-ketobutyrate treatment can significantly improve the exhaustion performance of old mice and increase the maintenance time of mice on the rotarod. α-ketobutyrate treatment also significantly improves the muscle index of the skeletal muscle of old mice, increases the number and cross-sectional area of muscle fibers, increases the number of muscle nuclei, and the expression of laminin B1 in the nuclear lamina of muscle nuclei. α-ketobutyrate treatment also increases the angiogenesis rate in the muscle tissue of old mice. Therefore, the endogenous small molecule metabolite α-ketobutyrate has good prospects in the preparation of drugs for preventing and / or treating skeletal muscle atrophy.

[0058] Finally, it should also be noted that the term "comprising", "including" or any other variation thereof is intended to cover non-exclusive inclusion, so that a process, method, article or device comprising a series of elements not only includes those elements but also includes other elements not expressly listed, or elements inherent to such process, method, article or device.

[0059] Although the preferred embodiments of the present invention have been described, additional changes and modifications can be made to these embodiments by those skilled in the art once they learn the basic creative concept. Therefore, the appended claims are intended to be construed as including the preferred embodiments as well as all changes and modifications falling within the scope of the present invention.

[0060] Obviously, those skilled in the art can make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, if these modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalent technologies, the present invention is also intended to include these modifications and variations.

Claims

1. Use of α-ketobutyrate in the preparation of a drug for preventing and / or treating skeletal muscle atrophy, characterized in that: The structural formula of the α-ketobutyrate is:

2. Use of α-ketobutyrate in the preparation of drugs for preventing and / or treating skeletal muscle aging.

3. Application of α-ketobutyrate in the preparation of drugs for improving exhaustion performance in elderly animals.

4. The use of α-ketobutyrate in the preparation of a drug for improving skeletal muscle performance in aged mice, characterized in that: The skeletal muscle performance of the aged mice includes the maintenance time of the aged mice on the rotarod.

5. The use of α-ketobutyrate in the preparation of a drug for improving the physiological characteristics of skeletal muscle in elderly animals, characterized in that: The skeletal muscle physiological characteristics include at least one of the tibialis anterior muscle index, the cross-sectional area of ​​muscle fibers, the rate of capillary formation in muscle tissue, the number of myocyte nuclei, and the expression of myocyte nuclear lamin LaminB1.

6. A drug for preventing and / or treating skeletal muscle atrophy, characterized in that: The active ingredient of the drug includes α-ketobutyrate.

7. A drug for preventing and / or treating skeletal muscle atrophy according to claim 6, characterized in that: The dosage form of the drug includes any one of tablets, capsules, oral liquids, granules, effervescent tablets and sustained-release preparations.

8. A drug for preventing and / or treating skeletal muscle aging, characterized in that: The active ingredient of the drug includes α-ketobutyrate.

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