Use of epimedium extract for preventing and treating sarcopenia

Abstract

The present invention discloses an epimedium extract for preventing and / or treating sarcopenia. The preparation method of the epimedium extract comprises the following steps: (1) weighing an appropriate amount of epimedium medicinal material, decocting twice with water, filtering, concentrating, and obtaining a first thick paste; (2) dissolving the first thick paste with water, standing, centrifuging, obtaining a supernatant, eluting the supernatant with a first solvent, a second solvent, and a third solvent respectively through a macroporous resin, collecting the third eluate, recovering the third solvent and concentrating, and obtaining a second thick paste; and (3) diluting the second thick paste with water, loading it on a preparative chromatographic column, eluting it with a mobile phase, collecting the first fraction and / or the fifth fraction at different time periods, concentrating, drying, and crushing to obtain the epimedium extract. The epimedium extract has a good improvement effect on sarcopenia.

Description

Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to use of an epimedium extract for preventing and treating sarcopenia. Background Art

[0002] Sarcopenia (abbreviated as "sarcopenia") is a common disease in middle-aged and elderly people. Its main clinical manifestation is the chronic and progressive reduction of muscle mass and quantity with aging, leading to loss of muscle strength [Dent E, Morley JE, Cruz-Jentoft AJ, et al. International Clinical Practice Guidelines for Sarcopenia (ICFSR): Screening, Diagnosis and Management. J Nutr Health Aging. 2018; 22 (10): 1148-1161.]. Sarcopenia can cause adverse consequences and is closely related to limb dysfunction, reduced self-care ability and significantly increased risk of fractures in the elderly, bringing great economic and social burdens [Cruz-Jentoft AJ, Sayer AA. Sarcopenia. Lancet. 2019 Jun 29; 393 (10191): 2636-2646.]. Studies have shown that human muscle reaches its peak at the age of 40, and then muscle mass gradually declines.The 2016 Asian Sarcopenia Management Guidelines estimated that the prevalence of sarcopenia in the elderly aged 60 to 70 years in China is around 4.1% to 11.5%. The prevalence of sarcopenia in men and women aged 70 years and above is 13.2% and 4.8%, respectively. The incidence rate increases significantly over 80 years old, up to 50% [Cruz-Jentoft AJ, Bahat G, Bauer J, Boirie Y, Bruyère O, Cederholm T, Cooper C, Landi F, Rolland Y, Sayer AA, Schneider SM, Sieber CC, Topinkova E, Vandewoud M, Visser M, Zamboni M; Writing Group for the European Working Group on Sarcopenia in Older People 2 (EWGSOP2), and the Extended Group for EWGSOP2. Sarcopenia: revised European consensus on definition and diagnosis. Age Ageing. 2019Jan 1; 48(1):16-31. Liu Juan, Ding Qingqing, Zhou Baiyu, Liu Xiang, Liu Jingmin, Liu Yongming, Ding Guoxian, Zhang Cuntai, Wang Jianye, Yu Pulin, Chinese Society of Geriatrics, Editorial Board of Chinese Journal of Geriatrics. Expert consensus on the diagnosis and treatment of sarcopenia in the elderly in China (2021) [J]. Chinese Journal of Geriatrics, 2021, 40(8):943-952.]. With the advent of an aging society, the incidence of sarcopenia is increasing year by year, seriously affecting the quality of life and health of the elderly. Therefore, it has attracted widespread attention from scholars at home and abroad, and extensive research has been carried out in the fields of risk factors, pathogenesis, and treatment of the disease. However, there are still differences in the understanding of the disease at home and abroad. In addition, since the cause of sarcopenia is still unknown, it is related to multiple factors and involves complex molecular mechanisms that are interrelated, which makes the treatment of the disease difficult. Currently, there is still a lack of a single effective therapeutic drug for sarcopenia. Resistance exercise is the main non-drug therapy for improving muscle mass and function. However, in reality, elderly patients have poor compliance with resistance exercise. Therefore, exploring effective treatment methods is an important direction in the current research field of sarcopenia treatment.

[0003] Traditional Chinese medicine has accumulated certain clinical experience in the treatment of sarcopenia. Traditional Chinese medicine classifies the clinical manifestations of thin and weak muscles as atrophy, which is similar to the clinical manifestations of sarcopenia, which are reduced muscle mass, decreased muscle strength or decreased physical function. Since sarcopenia and atrophy have highly overlapping connotations, traditional Chinese medicine often treats sarcopenia from the perspective of atrophy, and the effect is very good. Ancient Chinese medical literature records that the onset of atrophy is closely related to the spleen and kidney, among which the growth and aging of the human body are dominated by the rise and fall of kidney qi. As an aging-related geriatric syndrome, the onset of sarcopenia is more closely related to the deficiency and decline of kidney qi. "Suwen: Ancient Innocence" states: "At the age of eight, the kidney qi is strong, the hair grows and the teeth change; at the age of 28, the kidney qi is strong, the heavenly essence arrives, the essence and qi overflow, the yin and yang are in harmony, so the child can be born; at the age of 38, the kidney qi is balanced, the tendons and bones are strong, so the real teeth grow and grow to the maximum; at the age of 48, the tendons and bones are prosperous, and the muscles are full and strong; at the age of 58, the kidney qi is weak, the hair falls and the teeth become dry; at the age of 68, the yang qi is exhausted, the face becomes burnt, and the hair and temples become white; at the age of 78, the liver qi is weak, and the tendons cannot move; at the age of 88, the heavenly essence is exhausted, the essence is low, the kidney storage is weak, the body is at its extreme, and the teeth and hair fall." This shows that the growth and aging of the human body, including the fullness and strength of the muscles and the decline and atrophy, all change with the rise and fall of the kidney qi. As the kidney qi declines, the function of the muscles progressively declines, and the incidence of sarcopenia increases. According to epidemiological surveys, the prevalence of sarcopenia in the elderly is 10.8%-38.8% [Chen Shu, Yu Jing, Di Wenjuan, et al. Epidemiological survey of sarcopenia in the elderly in Nanjing community and analysis of influencing factors [J]. Practical Geriatrics, 2020, 34(08): 793-796. Che Yajie, Yang Xuanxuan, Zhang Ke, et al. Current status and related factors of sarcopenia in the elderly in Urumqi community [J]. Chinese Journal of Osteoporosis and Bone Mineral Diseases, 2020, 13(04): 304-310.], and the prevalence increases significantly with age, reaching 53.2% in the elderly aged 80 years and above [Meng P, Hu YX, Fan L, et al. Sarcopenia and sarcopenic obesity among menaged 80years and older in Beijing: prevalence and its association with functional performance [J]. Geriatr Gerontol Int. 2014, 14(1): 29-35.]. From this we can see that the "kidney" in traditional Chinese medicine is closely related to the onset of sarcopenia, and treating sarcopenia from the perspective of the kidney is highly feasible in traditional Chinese medicine theory.

[0004] There is no report in the prior art documents on the effect of the epimedium extract of the present invention in preventing and / or treating sarcopenia. Summary of the invention

[0005] Based on this, the present invention provides an epimedium extract for preventing and / or treating sarcopenia, and the preparation method of the epimedium extract comprises the following steps:

[0006] (1) weighing an appropriate amount of Epimedium medicinal material, adding water to decoct, filtering to obtain a first filtrate and a medicinal residue, adding water to decoct the medicinal residue, filtering to obtain a second filtrate, combining the first filtrate and the second filtrate, and concentrating to obtain a first thick paste;

[0007] (2) dissolving the first thick paste in water, allowing it to stand, centrifuging to obtain a supernatant, eluting the supernatant with a first solvent through a macroporous resin, discarding the first eluate, eluting with a second solvent, discarding the second eluate, eluting with a third solvent, collecting the third eluate, recovering the third solvent and concentrating it to obtain a second thick paste; and

[0008] (3) diluting the second thick paste with water, loading the sample onto a preparative chromatographic column, eluting with a mobile phase, collecting the first fraction and / or the fifth fraction at different time periods, concentrating, drying, and pulverizing to obtain the epimedium extract;

[0009] The first flow fraction is a flow fraction in the time period of 0 to 5 minutes;

[0010] The fifth fraction is a fraction in the time period of 20 to 40 minutes.

[0011] Furthermore, in step (1), the volume / mass (L / kg or ml / g) ratio of the water to the Epimedium medicinal material is 10-50, such as 20-40.

[0012] Furthermore, the decocting time is 0.5 to 5 hours, for example 1 to 3 hours.

[0013] Furthermore, the concentration is concentration under reduced pressure.

[0014] Furthermore, the relative density of the first thick paste at 55° C. to 70° C. is 1.08 to 1.18.

[0015] Furthermore, in step (2), the first thick paste is cooled and then dissolved in water, wherein the cooling is cooling to room temperature.

[0016] Furthermore, the mass / mass (kg / kg or g / g) ratio of the water to the Epimedium medicinal material is 0.1-2, for example, about 1.

[0017] Furthermore, the standing time is 1 to 10 hours, such as 3 to 5 hours.

[0018] Furthermore, the macroporous resin is a D101 macroporous resin.

[0019] Furthermore, the volume / mass (L / kg or ml / g) ratio of the supernatant to the macroporous resin is 0.1-2, such as about 0.4.

[0020] Furthermore, the first solvent is water, such as distilled water.

[0021] Furthermore, the first solvent is used for elution in an amount of 4 to 10 times the volume of the column bed, for example, 6 to 8 times the volume of the column bed.

[0022] Furthermore, the flow rate of the distilled water is 20-80 ml / min, for example, about 50 ml / min.

[0023] Furthermore, the second solvent is alcohol, such as ethanol.

[0024] Furthermore, the volume concentration percentage of the second solvent is 10% to 30%, for example, about 20%.

[0025] Furthermore, the second solvent is used for elution in an amount of 5 to 15 times the volume of the column bed, for example, 8 to 10 times the volume of the column bed.

[0026] Furthermore, the flow rate of the second solvent is 20-80 ml / min, for example, about 50 ml / min.

[0027] Furthermore, the third solvent is alcohol, such as ethanol.

[0028] Furthermore, the volume concentration percentage of the third solvent is 70% to 90%, for example, about 80%.

[0029] Furthermore, the third solvent is used for elution in an amount of 5 to 15 times the volume of the column bed, for example, 8 to 10 times the volume of the column bed.

[0030] Furthermore, the flow rate of the third solvent is 20-80 ml / min, for example, about 50 ml / min.

[0031] Furthermore, the relative density of the second thick paste at 55° C. to 70° C. is 1.08 to 1.18.

[0032] Furthermore, in step (3), the volume / volume ratio of the second thick paste to the water is 0.1-2, for example, about 1.

[0033] Furthermore, the conditions of the preparative chromatography are: using a chromatographic column whose filler is octadecylsilane bonded silica gel, mobile phase A is selected from one or more of acetonitrile, methanol and tetrahydrofuran, mobile phase B is water, and the gradient elution program is: 0-20 min, 25% A; 20-40 min, 40% A; 40-60 min, 48% A, the flow rate is 10-30 ml / min, the detection wavelength is 150-350 nm, and the column pressure is 10-30 MPa.

[0034] Furthermore, the specifications of the chromatographic column are: column length 600 mm, inner diameter 250 mm, particle size 10 μm.

[0035] Furthermore, phase A of the mobile phase is methanol.

[0036] Furthermore, the flow rate is 15-25 ml / min, for example, about 20 ml / min.

[0037] Furthermore, the detection wavelength is 230-270 nm, for example, 254 nm.

[0038] Furthermore, the column pressure is 15-25 MPa, for example, about 20 MPa.

[0039] According to another aspect of the present invention, there is provided a use of the above-mentioned Epimedium extract in the preparation of a medicament for preventing and / or treating sarcopenia.

[0040] According to another aspect of the present invention, there is provided a use of the above-mentioned epimedium extract in preparing a method for promoting myoblast differentiation in vitro.

[0041] According to another aspect of the present invention, there is provided a use of a pharmaceutical composition comprising the above-mentioned Epimedium extract in the preparation of a medicament for preventing and / or treating sarcopenia.

[0042] Furthermore, the pharmaceutical composition further comprises one or more drugs and / or extracts for preventing and / or treating sarcopenia.

[0043] Further, the drug is selected from one or more of the following: testosterone, selective androgen receptor modulators, growth hormone, ghrelin and myostatin inhibitors.

[0044] Furthermore, the pharmaceutical composition further comprises pharmaceutically acceptable excipients.

[0045] Furthermore, the auxiliary material is selected from one or more of the following: diluent, wetting agent, binder, disintegrant, inclusion agent, flavoring agent, sustained-release agent, retention agent, lubricant, dispersant, plasticizer, opacifier and antioxidant.

[0046] Furthermore, the dosage form of the pharmaceutical composition is powder, tablet, pill, capsule, film, lozenge, microcapsule, microsphere, granule, injection or oral solution.

[0047] Furthermore, the prevention and / or treatment of sarcopenia is manifested as one or more of the following: promoting myoblast differentiation, increasing muscle strength and muscle content, reducing body weight, body fat and bone mineral content, and delaying the aging process.

[0048] Beneficial effects of the present invention:

[0049] The epimedium extract obtained by the specific preparation method of the present invention can significantly promote myoblast differentiation, increase muscle strength and muscle content of SAMP8 sarcopenia model mice, reduce body weight, body fat and BMC content, and delay the aging process, and has no obvious effect on food intake, indicating that the epimedium extract can be used as a drug for the treatment of sarcopenia. BRIEF DESCRIPTION OF THE DRAWINGS

[0050] In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the drawings required for use in the description of the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without exceeding the scope of protection required by the present invention.

[0051] Figure 1 This is a schematic diagram of the effect of Epimedium Extract V on skeletal muscle of SAMP8 sarcopenia model mice. A is the HE staining result of tibialis anterior muscle; B is the DEXA result of body composition. DETAILED DESCRIPTION

[0052] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative work are within the scope of protection of the present invention.

[0053] Unless otherwise indicated, all technical and scientific terms and abbreviations used herein have the meanings commonly understood by those of ordinary skill in the field of the invention or the field in which the terms are used. Although any methods, conditions, substances or materials similar to or equivalent to those disclosed herein may be used in the practice of the present invention, preferred methods, conditions, substances or materials are described herein.

[0054] The present invention is intended to encompass all alternatives, variations and equivalents that may be included in the present invention as defined by the claims. Those skilled in the art will recognize many methods and materials similar or equivalent to those described herein that can be used in the practice of the present invention. The present invention is in no way limited to the methods and materials described.

[0055] As used in the specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise.

[0056] In the present invention, the term "comprising" is synonymous with "including". As used herein, the terms "comprising", "including", "having", "containing" or any other variation thereof are intended to cover a non-exclusive inclusion. For example, a composition, process, method, article, or apparatus comprising the listed elements is not necessarily limited to only those elements, but may include other elements not expressly listed or inherent to such composition, process, method, article, or apparatus.

[0057] As described in the background technology section, there is still a lack of a single effective therapeutic drug for sarcopenia. Resistance exercise is the main non-drug therapy for improving muscle mass and function, but in reality, elderly patients have poor compliance with resistance exercise, and there are no reports in the prior art documents on the effect of the epimedium extract claimed in the present invention on preventing and / or treating sarcopenia. In order to solve the above problems, the present invention provides an epimedium extract for preventing and / or treating sarcopenia, and the preparation method of the epimedium extract comprises the following steps:

[0058] (1) weighing an appropriate amount of Epimedium medicinal material, adding water to decoct, filtering to obtain a first filtrate and a medicinal residue, adding water to decoct the medicinal residue, filtering to obtain a second filtrate, combining the first filtrate and the second filtrate, and concentrating to obtain a first thick paste;

[0059] (2) dissolving the first thick paste in water, allowing it to stand, centrifuging to obtain a supernatant, eluting the supernatant with a first solvent through a macroporous resin, discarding the first eluate, eluting with a second solvent, discarding the second eluate, eluting with a third solvent, collecting the third eluate, recovering the third solvent and concentrating it to obtain a second thick paste; and

[0060] (3) diluting the second thick paste with water, loading the sample onto a preparative chromatographic column, eluting with a mobile phase, collecting the first fraction and / or the fifth fraction at different time periods, concentrating, drying, and pulverizing to obtain the epimedium extract;

[0061] The first flow fraction is a flow fraction in the time period of 0 to 5 minutes;

[0062] The fifth fraction is a fraction in the time period of 20 to 40 minutes.

[0063] In a preferred embodiment, in step (1), the volume / mass (L / kg or ml / g) ratio of the water to the Epimedium medicinal material is 10-50, such as 20-40.

[0064] In a preferred embodiment, the decocting time is 0.5 to 5 hours, such as 1 to 3 hours.

[0065] In a preferred embodiment, the concentration is concentration under reduced pressure.

[0066] In a preferred embodiment, the relative density of the first thick paste at 55° C. to 70° C. is 1.08 to 1.18.

[0067] In a preferred embodiment, in step (2), the first thick paste is cooled and then dissolved in water, wherein the cooling is cooling to room temperature.

[0068] In a preferred embodiment, the mass / mass (kg / kg or g / g) ratio of the water to the Epimedium medicinal material is 0.1-2, such as about 1.

[0069] In the present invention, "about" refers to a value within the range of ±5% of a specific value. For example, "about 1" includes ±5% of 1, or from 0.95 to 1.05.

[0070] In a preferred embodiment, the standing time is 1 to 10 hours, such as 3 to 5 hours.

[0071] In a preferred embodiment, the macroporous resin is a D101 macroporous resin.

[0072] In a preferred embodiment, the volume / mass (L / kg or ml / g) ratio of the supernatant to the macroporous resin is 0.1-2, such as about 0.4.

[0073] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 0.4" includes ±5% of 0.4, or from 0.38 to 0.42.

[0074] In a preferred embodiment, the first solvent is water, such as distilled water.

[0075] In a preferred embodiment, 4 to 10 times the volume of the column bed, such as 6 to 8 times the volume of the column bed, are used for elution with the first solvent.

[0076] In a preferred embodiment, the flow rate of the distilled water is 20-80 ml / min, such as about 50 ml / min.

[0077] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 50" includes ±5% of 50, or from 47.5 to 52.5.

[0078] In a preferred embodiment, the second solvent is an alcohol, such as ethanol.

[0079] In a preferred embodiment, the volume concentration percentage of the second solvent is 10% to 30%, for example, about 20%.

[0080] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 20" includes ±5% of 20, or from 19 to 21.

[0081] In a preferred embodiment, the second solvent is used for elution in an amount of 5 to 15 times the volume of the column bed, for example, 8 to 10 times the volume of the column bed.

[0082] In a preferred embodiment, the flow rate of the second solvent is 20-80 ml / min, such as about 50 ml / min.

[0083] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 50" includes ±5% of 50, or from 47.5 to 52.5.

[0084] In a preferred embodiment, the third solvent is an alcohol, such as ethanol.

[0085] In a preferred embodiment, the volume concentration percentage of the third solvent is 70% to 90%, for example, about 80%.

[0086] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 80" includes ±5% of 80, or from 76 to 84.

[0087] In a preferred embodiment, the third solvent is used for elution in an amount of 5 to 15 times the volume of the column bed, for example, 8 to 10 times the volume of the column bed.

[0088] In a preferred embodiment, the flow rate of the third solvent is 20-80 ml / min, such as about 50 ml / min.

[0089] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 50" includes ±5% of 50, or from 47.5 to 52.5.

[0090] In a preferred embodiment, the relative density of the second thick paste at 55° C. to 70° C. is 1.08 to 1.18.

[0091] In a preferred embodiment, in step (3), the volume / volume ratio of the second thick paste to the water is 0.1-2, for example, about 1.

[0092] In the present invention, "about" refers to a value within the range of ±5% of a specific value. For example, "about 1" includes ±5% of 1, or from 0.95 to 1.05.

[0093] In a preferred embodiment, the conditions of the preparative chromatography are: a chromatographic column with octadecylsilane bonded silica gel as filler is used, the mobile phase A is selected from one or more of acetonitrile, methanol and tetrahydrofuran, the mobile phase B is water, the gradient elution program is: 0-20 min, 25% A; 20-40 min, 40% A; 40-60 min, 48% A, the flow rate is 10-30 ml / min, the detection wavelength is 150-350 nm, and the column pressure is 10-30 MPa.

[0094] In a preferred embodiment, the specifications of the chromatographic column are: column length 600 mm, inner diameter 250 mm, and particle size 10 μm.

[0095] In a preferred embodiment, phase A of the mobile phase is methanol.

[0096] In a preferred embodiment, the flow rate is 15-25 ml / min, such as about 20 ml / min.

[0097] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 20" includes ±5% of 20, or from 19 to 21.

[0098] In a preferred embodiment, the detection wavelength is 230-270 nm, for example 254 nm.

[0099] In a preferred embodiment, the column pressure is 15-25 MPa, such as about 20 MPa.

[0100] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 20" includes ±5% of 20, or from 19 to 21.

[0101] According to another aspect of the present invention, there is provided a use of the above-mentioned Epimedium extract in the preparation of a medicament for preventing and / or treating sarcopenia.

[0102] According to another aspect of the present invention, there is provided a use of the above-mentioned epimedium extract in preparing a method for promoting myoblast differentiation in vitro.

[0103] According to another aspect of the present invention, there is provided a use of a pharmaceutical composition comprising the above-mentioned Epimedium extract in the preparation of a medicament for preventing and / or treating sarcopenia.

[0104] In a preferred embodiment, the pharmaceutical composition further comprises one or more drugs and / or extracts for preventing and / or treating sarcopenia.

[0105] In a preferred embodiment, the drug is selected from one or more of the following: testosterone, selective androgen receptor modulators, growth hormone, ghrelin and myostatin inhibitors.

[0106] In a preferred embodiment, the pharmaceutical composition further comprises a pharmaceutically acceptable excipient.

[0107] In a preferred embodiment, the pharmaceutical composition of the present invention contains at least one pharmaceutically acceptable excipient in an amount of 0.00001 to 50 wt.%, or 0.0001 to 10 wt.%, or 0.0001 to 5 wt.%, or 0.005 to 1 wt.%, or 0.1 to 20 wt.%, or 0.5 to 15 wt.%, or 1 to 5 wt.%, based on the weight of the pharmaceutical composition.

[0108] In the present invention, the term "pharmaceutically acceptable" refers to a substance, such as a carrier or diluent, that does not abrogate the biological activity or properties of the compound and is relatively non-toxic, i.e., a substance that does not cause undesirable biological effects or interact in a deleterious manner with any of its components when administered to an individual.

[0109] In the present invention, the term "pharmaceutically acceptable excipient" refers to carriers and / or excipients that are pharmacologically and / or physiologically compatible with subjects and active ingredients (i.e., capable of inducing the desired therapeutic effect without causing any undesirable local or systemic effects), which are well known in the art (see, for example, Remington's Pharmaceutical Sciences. Edited by Gennaro AR, 19th ed. Pennsylvania: Mack Publishing Company, 1995).

[0110] In a preferred embodiment, the auxiliary material is selected from one or more of the following: diluents, excipients, excipients, wetting agents, binders, disintegrants, inclusion agents, flavoring agents, sustained-release agents, retention agents, lubricants, dispersants, plasticizers, opacifiers and antioxidants.

[0111] Those skilled in the art will know how to select specific chemical substances within the scope of the above-mentioned auxiliary material category.For example, the diluent can be selected from one or more of the following: icing sugar, starch, compressible starch, lactose, dextrin, mannitol, sorbitol, microcrystalline cellulose, calcium sulfate and calcium carbonate.The wetting agent can be selected from one or more of the following: polyoxymethylene stearate, poloxamer, polyoxyethylene sorbitan fatty acid ester, polyoxyethylene hydrogenated castor oil, polyoxyethylene alkyl ether, polysorbate such as polysorbate 80, cetyl alcohol, glycerol fatty acid ester (such as triacetin, glyceryl monostearate and the like), polyoxyethylene fatty acid ester, polyethylene glycol fatty acid ester, sodium lauryl sulfate, sorbitol fatty acid ester, sucrose fatty acid ester, polyoxyethylene ether, benzalkonium chloride, polyoxyethylene castor oil and docusate sodium.The adhesive can be selected from one or more of the following: polyvinyl pyrrolidone, hydroxypropyl cellulose, polyethylene glycol and methylcellulose. The disintegrant may be selected from one or more of the following: carboxymethylcellulose, carboxymethylcellulose calcium salt and sodium carboxymethylcellulose. The flavoring agent may be selected from one or more of the following: sorbitol, glucose, mannose, sucrose and lactose. The dispersant may be selected from one or more of the following: cross-linked sodium carboxymethylcellulose, sodium starch glycolate and pre-gelatinized corn starch. The plasticizer may be dibutyl sebacate and / or various citrates. The sustained-release agent may be selected from one or more of the following: sodium carboxymethylcellulose, low-substituted hydroxypropylcellulose, hydroxypropyl methylcellulose, microcrystalline cellulose, hydroxyethylcellulose, gum arabic, gelatin and shellac. The antioxidant may be selected from one or more of the following: sodium bisulfite, sodium pyrosulfite, sodium sulfite and sodium thiosulfate. The lubricant may be selected from one or more of the following: calcium stearate, talc, magnesium stearate, stearic acid and colloidal silicon dioxide.

[0112] These excipients are preferably pharmaceutically inert, or may have a synergistic or additive effect to enhance the therapeutic activity of the pharmaceutical composition, and the above-mentioned excipients are only listed. The excipients actually used in the present invention are not limited to the above-mentioned excipients and can be adjusted according to actual conditions to achieve the effects of the present invention.

[0113] In a preferred embodiment, the pharmaceutical composition is in the form of powder, tablet, pill, capsule, film, lozenge, microcapsule, microsphere, granule, injection or oral solution.

[0114] In a preferred embodiment, the prevention and / or treatment of sarcopenia is manifested as one or more of the following: promoting myoblast differentiation, increasing muscle strength and muscle content, reducing body weight, body fat and bone mineral content, and delaying the aging process.

[0115] According to another aspect of the present invention, there is provided the above-mentioned Epimedium extract or the above-mentioned pharmaceutical composition for preventing and / or treating sarcopenia in a subject.

[0116] According to another aspect of the present invention, there is provided a method for preventing and / or treating sarcopenia in a subject, comprising administering an effective amount of the above-mentioned Epimedium extract or the above-mentioned pharmaceutical composition to the subject.

[0117] In the present invention, the term "subject" is a mammal. The mammal may be a human, a non-human primate, a mouse, a rat, a dog, a cat, a horse or a cow, but is not limited to these examples. Mammals other than humans can be advantageously used as subjects representing sarcopenia models. Preferably, the subject is a human.

[0118] The "effective amount" of the pharmaceutical composition or preparation used in the present invention can obtain the desired treatment and / or prevention effect. The amount effective for this purpose will depend on, for example, the pharmaceutical composition, the mode of administration, the stage and severity of the disease to be treated, the individual's body weight and overall health, and the judgment of the prescribing physician. The dosage can be given once a week, or once every two days or every day, or even several times a day. The dosage unit can be given in a short period of time (e.g., weeks to months) or longer time periods (months to years).

[0119] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention. The experimental methods in the following examples without specifying specific conditions are usually carried out according to conventional conditions or conditions recommended by the manufacturer.

[0120] Unless otherwise defined, all professional and scientific terms used herein have the same meanings as those familiar to those skilled in the art. In addition, any methods and materials similar or equivalent to those described herein can be applied to the method of the present invention. The preferred implementation methods and materials described herein are for demonstration purposes only. The above-mentioned features mentioned in the present invention, or the features mentioned in the embodiments, can be combined arbitrarily. All features disclosed in this patent specification can be used in combination with any form of composition, and each feature disclosed in the specification can be replaced by any alternative feature that can provide the same, equal or similar purpose. Therefore, unless otherwise specified, the disclosed features are only general examples of equal or similar features.

[0121] Example

[0122] Preparation Example

[0123] 1. Experimental Materials

[0124] D101 macroporous resin was purchased from Sinopharm Chemical Reagent Co., Ltd. DMEM high glucose medium and horse serum were purchased from Gibco. Methanol and acetonitrile for HPLC were chromatographically pure and were purchased from Thermo Fisher; Trizol reagent (batch number: G826KA6730), DEPC water and primers were purchased from Shanghai Sangon Biotechnology Co., Ltd. Chloroform, formaldehyde, anhydrous ethanol and sodium carboxymethyl cellulose (CMC-Na) were purchased from Sinopharm Chemical Reagent Co., Ltd. Isopropanol was purchased from Merck, Germany. RNA reverse transcription kit was purchased from Applied Biological Materials, Canada. Green ProTaq HS premixed qPCR kit (including ROX) was purchased from Hunan Aikerui Bioengineering Co., Ltd. TNF-α and IFN-γ factors were purchased from Shanghai Biyuntian Biotechnology Co., Ltd., and Epimedium was purchased from Shanghai Kangqiao Decoction Pieces Factory, which met the standards of the 2020 edition of the Chinese Pharmacopoeia. Water was ultrapure water, and other reagents were analytical grade.

[0125] 2. Experimental methods

[0126] Take the herbal medicine of Epimedium, add 20 to 40 times the volume of the herbal medicine in water, decoct for 1 to 3 hours, filter, and set aside the filtrate; add 20 to 40 times the volume of the herbal medicine in water to decoct for 1 to 3 hours, filter, and set aside the filtrate; combine the filtrates obtained from the two extractions, and concentrate under reduced pressure to a thick paste 1 with a relative density of d=1.08 to 1.18 (55 to 70°C), cool (to room temperature), add water of the same mass as the herbal medicine to the thick paste 1 to dissolve it, let it stand at room temperature for about 3 to 5 hours, centrifuge, absorb the supernatant, and pass it through a D101 macroporous resin column (supernatant: resin = 1:2.5), eluted with distilled water (6 to 8 times the volume of the column bed), the flow rate is 50ml / min, the water eluent is discarded, and then eluted with 20% ethanol (8 to 10 times the volume of the column bed), the eluent is collected, and then eluted with 80% ethanol (8 to 10 times the volume of the column bed), the eluent is collected, and the 80% ethanol is recovered and concentrated to a thick paste 2 with a relative density of d = 1.08 to 1.18 (55 to 70°C); the thick paste 2 is diluted with an equal volume of water, loaded, and subjected to preparative chromatography (filler C 18 , 250×600mm, 10μm), the mobile phase A of the preparative chromatography is methanol, the phase B is water, the flow rate is 20mL / min; the detection wavelength is 254nm; the column pressure is 20Mpa; the elution program is: the volume fraction of the mobile phase A from 0 to 20min is 25%; the volume fraction of the mobile phase A from 20min to 40min is 40%; the volume fraction of the mobile phase A from 40min to 60min is 48%.

[0127] The fractions were collected at the following time periods:

[0128] F1: 0-5min; F2: 5-10min; F3: 10-15min; F4: 15-20min; F5: 20-25min; F6: 25-30min; F7: 30-35min; F8: 35-40min; F9: 40-45min; F10: 45-50min; F11: 50-55min; F12: 55-60min. The fractions F5-8 and F9-12 were combined to obtain 6 fractions, which were concentrated, dried and pulverized to obtain 6 kinds of extract powders, which were named extracts I (F1), II (F2), III (F3), IV (F4), V (F5-8) and VI (F9-12) in sequence.

[0129] Pharmacodynamics Examples

[0130] Experiment 1: Effects of Epimedium Extract on Myoblasts

[0131] 1. Experimental purpose

[0132] The effects of different extracts of Epimedium on the myogenic differentiation of C2C12 cells were evaluated by examining the effect of drugs inducing myoblast differentiation.

[0133] 2. Experimental methods

[0134] 2.1 C2C12 cell induction and myogenic differentiation

[0135] When the C2C12 myoblasts grew and merged to 70% to 80%, the growth medium was discarded and serum-free medium was added for starvation overnight. Then, the differentiation medium (DMEM high glucose medium + 2% horse serum + double antibody) was replaced to induce muscle differentiation for 6 days to form myotubes. New differentiation medium was replaced every 1 day and culture was continued.

[0136] 2.2 Cell grouping and drug administration

[0137] C2C12 myoblasts were randomly divided into control group, TNF-α / IFN-γ stimulation group, and epimedium extract I-VI (10, 50μM), with 3 replicates in each group. The control group was cultured with growth medium for 24h; the stimulation group was cultured with growth medium containing 10ng / mL TNF-α and 10ng / mL IFN-γ for 24h; the epimedium extract I-VI group was cultured with growth medium containing 10ng / mL TNF-α, 10ng / mL IFN-γ and epimedium extract I-VI (10, 50μM) for 24h. On the 5th day of induction of myogenic differentiation, each group was cultured with differentiation medium containing or without drug for 24h, and the morphology of myotube cells in each group was observed in real time under a fluorescent inverted microscope and photographed.

[0138] 2.3 CCK8 assay to detect cell viability

[0139] C2C12 myoblasts in logarithmic growth phase were cultured at 5×10 3 The cells were inoculated in a 96-well plate at a density of 1000 μM and cultured overnight until they adhered to the wall. Different concentrations of Epimedium extracts I-VI (0, 10, 20, 50, 100 μM) were added and incubated for 24 h. 20 μL of CCK8 reagent was added to each well and cultured for 1 h. The optical density (OD) value of each well was detected at a wavelength of 450 nm using an ELISA reader, and the cell viability was calculated. Three replicate wells were set up for each group.

[0140] 2.4 RT-qPCR detection of mRNA expression of MRFs family factors in myoblast-differentiated myotubes

[0141] The cells were treated according to the method under "2.2", myoblasts and differentiated myotubes were collected, RZ lysis buffer was added, RNA was extracted using a total RNA extraction kit and reverse transcribed to obtain cDNA, and the mRNA expression of inflammatory factors in myoblasts and the mRNA expression of MRFs family factors in differentiated myotubes were detected. GAPDH was used as an internal reference and the 2-ΔΔCT method was used for analysis. The primer sequences were derived from PrimerBank (as shown in Table 1) and synthesized by Sangon Biotech (Shanghai) Co., Ltd.

[0142] Table 1 Primer sequences

[0143]

[0144]

[0145] 3. Experimental results

[0146] 3.1 Effects of different extracts of Epimedium on the viability of C2C12 myoblasts

[0147] The results are shown in Table 2. Compared with the control group, 100 μM of different extracts of Epimedium had an inhibitory effect on the survival rate of C2C12 myoblasts (P<0.001), so concentrations below 50 μM were selected for subsequent experiments.

[0148] Table 2 Effects of different extracts of Epimedium on the viability of C2C12 myoblasts

[0149]

[0150]

[0151] 3.2 Effects of different extracts of Epimedium on the morphology of differentiated myotubes and the expression of MRFs genes

[0152] The results are shown in Table 3. Compared with the control group, the nuclear fusion of myotubes in the stimulation group was reduced, the morphology was atrophied, and the diameter and length were reduced. The nuclear fusion of myotubes in the 50 μM epimedium extract I and V was increased, the morphology was normal, and the diameter and length did not change significantly. The improvement effect of epimedium extract V was the most significant; epimedium extracts II, III, IV, and VI had no improvement effect.

[0153] Compared with the control group, the mRNA expressions of MyoD, Mrf5, Mygenin and Myf6 in the stimulation group were decreased (P<0.05, P<0.01); compared with the stimulation group, 10μM and 50μM epimedium extract V could significantly increase the mRNA expressions of MyoD, Mrf5, Mygenin and Myf6 (P>0.05), and 50μM epimedium extract I could increase the mRNA expressions of MyoD and Mrf5, while other extracts had no effect on the mRNA expressions of MyoD, Mrf5, Mygenin and Myf6.

[0154] Table 3 Effects of different extracts of Epimedium on the expression of MyoD, Mrf5, Mygenin and Myf6 mRNA in C2C12 myoblasts

[0155]

[0156]

[0157] Note: * indicates that compared with the stimulation group, P<0.05; ** indicates that compared with the stimulation group, P<0.01; *** indicates that compared with the stimulation group, P<0.001;

[0158] 4. Conclusion

[0159] The results of cell experiments showed that Epimedium Extract V could significantly promote myoblast differentiation and had the best effect.

[0160] Experiment 2: Effects of Epimedium Extract V on SAMP8 Sarcopenia Model Mice

[0161] 1. Experimental Materials

[0162] 1.1 Experimental animals

[0163] SAMP8 male mice, 16 weeks old, weighing 30±2 g; SAMR1 male mice, 16 weeks old, weighing 30±2 g were purchased from Jiangsu Ai Ling Fei Biotechnology Co., Ltd.

[0164] The experimental animals were kept in the SPF animal room of Shanghai Experimental Animal Center, with a temperature of (24±1)℃, a relative humidity of 50% to 70%, and a light / dark cycle of 12h / 12h. The experimental operation procedures strictly followed the relevant regulations and provisions of the Ethics Committee of Shanghai Experimental Animal Center.

[0165] 1.2 Experimental drugs

[0166] The epimedium extract V used in the embodiment of the present invention is homemade in the laboratory.

[0167] 1.3 Experimental instruments

[0168] Dual-energy X-ray absorptiometry (iNSiGHT VET DXA, OsteoSys, KOREAN); YLS-13A muscle strength tester, Jinan Yiyan Technology Co., Ltd.

[0169] 2 Methods

[0170] 2.1 Grouping and Dosing

[0171] After 1 week of adaptive feeding with ordinary diet, 32 SAMP8 mice were randomly divided into a model group, a low-dose group (5 mg / kg), a medium-dose group (10 mg / kg), and a high-dose group (20 mg / kg) of Epimedium Extract V, with 8 mice in each group; 8 SAMR1 mice of the same age were used as a negative control group.

[0172] Epimedium extract V was dissolved in 0.3% sodium carboxymethylcellulose solution. Each treatment group was given different concentrations of Epimedium extract V solution by gavage for 12 consecutive weeks, once a day, with a gavage volume of 0.1mL / 10g. The negative control group and the model group were given 0.3% sodium methylcellulose solution by gavage for 12 consecutive weeks, once a day, with a gavage volume of 0.1mL / 10g.

[0173] 2.2 Detection indicators and methods

[0174] 2.2.1 General determination

[0175] The body weight and food intake of mice were measured according to conventional methods.

[0176] Method for determination of mouse weight and food intake: The weight of mice was determined according to the standard operating procedure for weighing. Experimental equipment: electronic balance (range: 0-200g, sensitivity 0.01g). Operation steps: Select a suitable height and stable position to place the electronic scale; plug in the power supply, turn on the machine, and after the electronic balance is ready, put a weighing box of appropriate size on it and adjust it to zero. According to the "Operational Procedures for Grasping and Fixing Experimental Animals", grab the mouse from the feeding box, put it in the weighing box, and read the reading after the value stabilizes. If the value keeps jumping, select the middle value and record it. The read value is recorded in a dedicated original experimental record sheet. Put the mouse back in the cage, cover the cage lid, put it back in the cage rack, and plug in the drinking bottle. After weighing, put the mouse cage neatly, turn off the balance, disconnect the power supply, wipe the electronic balance clean and put it back in place, and fill in the use record of the electronic balance.

[0177] The food intake of mice was measured using the rat and mouse diet and water intake monitoring system. The XR1130 rat and mouse diet and water intake monitoring system is a system used to regularly measure the food intake behavior of rats and mice raised in a single laboratory. It was purchased from Shanghai Xinruan Information Technology Co., Ltd. The measurement time is set to weekly. Through regular measurements, the software records the food intake of each activity cage. Food can be added midway, and the data can automatically accumulate the amount of food in the trough. The data dashboard can monitor data such as the amount of food (grams). Mouse feed was purchased from the Shanghai Laboratory Animal Research Center and produced in accordance with the standardized quality requirements of laboratory animal compound feed.

[0178] 2.2.2 Aging score

[0179] The aging rating scale was used to evaluate the degree of apparent aging, which includes reactive, passive, skin and hair, eye and spine-related aging, and is divided into 11 categories. Each category is scored from 0 to 4 points and is evaluated every 0, 4, 8, and 12 weeks.

[0180] 2.2.3 Mouse muscle strength measurement

[0181] The forelimb grip strength of mice was measured using a YLS-13A muscle strength tester (Jinan Yiyan Technology Co., Ltd., Jinan, China). The mice were lifted and fixed by their tails so that their limbs could grasp a wire mesh. Then, the mice were gently pulled back by their tails, with their posture parallel to the surface of the table, until they released the mesh. The peak force exerted by the mouse limbs was recorded in grams (g). Evaluations were performed every 0, 4, 8, and 12 weeks, and each mouse was tested three times, and the average was taken and weighted for statistical analysis.

[0182] 2.2.4 Body composition of mice

[0183] Three days before the mice were killed, dual-energy X-ray absorptiometry (DEXA) was used to measure body composition, including body fat content, muscle content, bone mineral content, etc. The data were analyzed using the software provided by the manufacturer.

[0184] 2.2.5 Skeletal muscle pathology detection

[0185] After the intervention, the mice were killed and the tibialis anterior muscle was obtained. The tibialis anterior muscle was fixed with 4% paraformaldehyde for 24 hours, dehydrated, paraffin-embedded, and cut into 5 μm paraffin sections. For skeletal muscle tissue sections, HE staining was used to detect the morphology and number of mouse skeletal muscle fibers.

[0186] 2.2.6 Statistical methods

[0187] The experimental data were analyzed using SPSS 21.0 software. The measurement data were expressed as mean ± standard deviation. One-way ANOVA was used for comparison among the groups. P < 0.05 was considered statistically significant.

[0188] 3 Results

[0189] 3.1 Effect of Epimedium Extract V on Food Intake in SAMP8 Sarcopenia Model Mice

[0190] The results are shown in Table 4. Four weeks after administration, the average food intake of the low-dose, medium-dose and high-dose groups of Epimedium Extract V was greater than that of the model group and the negative control group; 8 weeks after administration, the food intake of the model group and the negative control group increased compared with that 4 weeks after administration, and the food intake of the low-dose, medium-dose and high-dose groups of Epimedium Extract V decreased compared with before; 12 weeks after administration, compared with the model group, the food intake of the low-dose group increased (P < 0.05), indicating that Epimedium Extract V had no significant effect on the food intake of SAMP8 sarcopenia model mice.

[0191] Table 4 Effect of Epimedium extract V on food intake in SAMP8 sarcopenia model mice

[0192]

[0193] 3.2 Effect of Epimedium Extract V on Body Weight of SAMP8 Sarcopenia Model Mice

[0194] The results are shown in Table 5. There was no significant difference in body weight among the groups before administration, indicating that the groups were comparable. There was no significant difference in the body weight of mice among the groups 4 weeks after administration. The average body weight of mice in the model group and the low-dose group of Epimedium Extract V increased compared with before, and the body weight of mice in the other groups decreased compared with before. There was no significant difference in the body weight of mice among the groups 8 weeks after administration, and the body weight of mice was basically the same as 4 weeks after administration. After 12 weeks of administration, compared with the model group, the body weight of the medium-dose group and the negative control group was significantly reduced (P < 0.01), and the body weight of the high-dose group was significantly reduced (P < 0.05), indicating that Epimedium Extract V may have a certain weight loss effect on the body weight of SAMP8 sarcopenia model mice.

[0195] Table 5 Effect of Epimedium Extract V on Body Weight of SAMP8 Sarcopenia Model Mice

[0196]

[0197] Note: Compared with the model group, *P<0.05, **P<0.01.

[0198] 3.3 Effect of Epimedium Extract V on Aging in SAMP8 Sarcopenia Model Mice

[0199] The results are shown in Table 6. There was no significant difference in the aging scores of SAMP8 mice among the groups before administration, indicating that the groups were comparable. After 4 weeks of administration, compared with the model group, the aging scores of the medium-dose group and the high-dose group decreased (P < 0.05), and the aging score of the negative control group decreased significantly (P < 0.01); compared with the low-dose group, the aging scores of the medium-dose group and the high-dose group decreased (P < 0.05). After 8 weeks of administration, compared with the model group, the aging scores of the medium-dose group and the high-dose group decreased (P < 0.05), and the aging score of the negative control group decreased significantly (P < 0.01); compared with the low-dose group, the aging score of the medium-dose group decreased (P < 0.05), and the aging score of the high-dose group showed a decreasing trend. After 12 weeks of administration, compared with the model group, the aging score of the medium-dose group was reduced (P < 0.05), and the aging score of the negative control group was significantly reduced (P < 0.01); compared with the low-dose group, the aging scores of the medium-dose group and the high-dose group tended to decrease.

[0200] Table 6 Effect of Epimedium Extract V on the aging degree of SAMP8 sarcopenia model mice

[0201]

[0202]

[0203] Note: Compared with the model group, *P<0.05, **P<0.01; compared with the low-dose group, △ P<0.05, △△ P<0.01.

[0204] 3.4 Effect of Epimedium Extract V on Muscle Strength in SAMP8 Sarcopenia Model Mice

[0205] The results are shown in Table 7. Before administration, compared with the negative control group, the muscle strength of each group decreased significantly (P < 0.01); there was no significant difference in the muscle strength of SAMP8 mice among the groups, indicating that the experimental system was reliable and the groups were comparable. After 4 weeks of administration, compared with the model group, the muscle strength of the medium-dose group, the high-dose group and the negative control group increased significantly (P < 0.01); compared with the low-dose group, the muscle strength of the medium-dose group and the high-dose group increased (P < 0.05). After 8 weeks of administration, the muscle strength of the medium-dose group, the high-dose group and the negative control group increased significantly (P < 0.01). After 12 weeks of administration, compared with the model group, the muscle strength of the high-dose group increased (P < 0.05), and the muscle strength of the medium-dose group and the negative control group increased significantly (P < 0.01); compared with the low-dose group, the muscle strength of the medium-dose group increased (P < 0.05), and the muscle strength of the high-dose group had an increasing trend.

[0206] Table 7 Effects of Epimedium Extract V on Muscle Strength in SAMP8 Sarcopenia Model Mice

[0207]

[0208] Note: Compared with the model group, *P<0.05, **P<0.01; compared with the low-dose group, △ P<0.05, △△ P<0.01.

[0209] 3.5 Effects of Epimedium Extract V on Body Fat, Muscle and Bone Mineral Content (BMC) in SAMP8 Sarcopenia Model Mice

[0210] The results after 12 weeks of administration are shown in Table 8. Compared with the model group, the body fat content of the medium-dose group was significantly reduced, the muscle content was significantly increased (P < 0.01), and the BMC was reduced (P < 0.05); the body fat content of the negative control group was reduced, the muscle content was increased (P < 0.05), and the BMC had a downward trend; the BMC and body fat content of the high-dose group had a downward trend, and the muscle content had an upward trend. Figure 1 As shown in B.

[0211] Table 8 Effects of Epimedium Extract V on body fat content, muscle content and bone mineral content in SAMP8 sarcopenia model mice

[0212]

[0213] Note: Compared with the model group, *P<0.05, **P<0.01.

[0214] 3.6 Effects of Epimedium Extract V on Skeletal Muscle Sections of SAMP8 Sarcopenia Model Mice

[0215] HE-stained sections were used to analyze the pathological damage of the skeletal muscle of SAMP8 mice. The skeletal muscle tissue of normal SAMR1 mice is arranged intact, regular and clear. However, in the SAMP8 mouse model, the skeletal muscle is abnormal and uneven, with a smaller cross-sectional area and wider intermuscular space. However, total flavonoids from Epimedium significantly reversed these changes. After intervention with total flavonoids from Epimedium, the muscle fiber morphology of mice was improved compared with the model group, the arrangement was neater than before, the intermuscular space was smaller than before, and it had a larger cross-sectional area, indicating that total flavonoids from Epimedium can help restore the pathological damage of skeletal muscle in aging mice, and the medium-dose group performed particularly well. SAMP8 mouse tibialis anterior muscle section as shown Figure 1 As shown in A.

[0216] 4 Summary

[0217] Epimedium extract V can increase the muscle strength and muscle content of SAMP8 sarcopenia model mice, reduce body weight, body fat and BMC content, and delay the aging process, without significantly affecting the food intake, indicating that Epimedium extract V can be used as a drug for the treatment of sarcopenia.

[0218] The embodiments of the present invention are described in detail above. Specific examples are used herein to illustrate the principles and implementation methods of the present invention. The description of the above embodiments is only used to help understand the method of the present invention and its core idea. At the same time, changes or deformations made by those skilled in the art based on the ideas of the present invention, the specific implementation methods and application scope of the present invention, all belong to the scope of protection of the present invention. In summary, the content of this specification should not be understood as limiting the present invention.

Claims

1. An epimedium extract for preventing and / or treating sarcopenia, characterized in that: The preparation method of the epimedium extract comprises the following steps: (1) weighing an appropriate amount of Epimedium medicinal material, decocting with water, filtering to obtain a first filtrate and a medicinal residue, decocting the medicinal residue with water, filtering to obtain a second filtrate, combining the first filtrate and the second filtrate, and concentrating to obtain a first thick paste; (2) adding water to dissolve the first thick paste, standing, centrifuging to obtain a supernatant, passing the supernatant through a macroporous resin, eluting with a first solvent for 6 to 8 times the column bed volume, discarding the first eluent, eluting with a second solvent for 8 to 10 times the column bed volume, discarding the second eluent, eluting with a third solvent for 8 to 10 times the column bed volume, collecting the third eluent, recovering the third solvent and concentrating it to obtain a second thick paste, wherein the first solvent is water, the second solvent is 19% to 21% ethanol, and the third solvent is 76% to 84% ethanol; and (3) diluting the second thick paste with water, loading the sample onto a preparative chromatographic column, eluting with a mobile phase, collecting the first fraction and / or the fifth fraction at different time periods, concentrating, drying, and pulverizing to obtain the epimedium extract; The conditions for the preparative chromatography are as follows: a chromatographic column with octadecylsilane bonded silica as filler is used, the specifications of the chromatographic column are as follows: column length 600 mm, inner diameter 250 mm, particle size 10 μm, mobile phase A is methanol, mobile phase B is water, gradient elution program is: 0-20 min, 25% A; 20-40 min, 40% A; 40-60 min, 48% A, flow rate is 19 ml / min-21 ml / min, detection wavelength is 150-350 nm, column pressure is 19 Mpa-21 Mpa; Wherein, the first flow fraction is the flow fraction of the time period of 0 to 5 minutes; Wherein, the fifth fraction is the fraction in the time period of 20 to 40 minutes.

2. The epimedium extract according to claim 1, characterized in that In step (1), the volume / mass ratio of the water to the Epimedium medicinal material is 10 to 50, with the unit of L / kg or ml / g.

3. The epimedium extract according to claim 2, characterized in that The volume / mass ratio of the water to the epimedium medicinal material is 20-40, with the unit of L / kg or ml / g.

4. The epimedium extract according to claim 1, characterized in that In step (1), the decocting time is 0.5 to 5 hours.

5. The epimedium extract according to claim 4, characterized in that The decoction time is 1 to 3 hours.

6. The epimedium extract according to claim 1, characterized in that In step (1), the concentration is concentration under reduced pressure.

7. The epimedium extract according to claim 1, characterized in that In step (1), the relative density of the first thick paste at 55° C. to 70° C. is 1.08 to 1.

18.

8. The epimedium extract according to claim 1, characterized in that In step (2), the first thick paste is cooled and then dissolved in water, wherein the cooling is cooling to room temperature.

9. The epimedium extract according to claim 1, characterized in that In step (2), the mass / mass ratio of the water to the Epimedium medicinal material is 0.1 to 2, with the unit of kg / kg or g / g.

10. The epimedium extract according to claim 9, characterized in that The mass / mass ratio of the water to the Epimedium medicinal material is 0.95-1.05, with the unit of kg / kg or g / g.

11. The epimedium extract according to claim 1, characterized in that In step (2), the standing time is 1 to 10 hours.

12. The epimedium extract according to claim 11, characterized in that The standing time is 3 to 5 hours.

13. The epimedium extract according to claim 1, characterized in that In step (2), the macroporous resin is a D101 macroporous resin.

14. The epimedium extract according to claim 1, characterized in that In step (2), the volume / mass ratio of the supernatant to the macroporous resin is 0.1-2, in units of L / kg or ml / g.

15. The epimedium extract according to claim 14, characterized in that The volume / mass ratio of the supernatant to the macroporous resin is 0.38-0.42, with the unit of L / kg or ml / g.

16. The epimedium extract according to claim 1, characterized in that In step (2), the first solvent is distilled water.

17. The epimedium extract according to claim 16, characterized in that The flow rate of the distilled water is 20-80 ml / min.

18. The epimedium extract according to claim 17, characterized in that The flow rate of the distilled water is 47.5 ml / min to 52.5 ml / min.

19. The epimedium extract according to claim 1, characterized in that In step (2), the flow rate of the second solvent is 20 to 80 ml / min.

20. The epimedium extract according to claim 19, characterized in that The flow rate of the second solvent is 47.5 ml / min to 52.5 ml / min.

21. The epimedium extract according to claim 1, characterized in that In step (2), the flow rate of the third solvent is 20 to 80 ml / min.

22. The epimedium extract according to claim 21, characterized in that The flow rate of the third solvent is 47.5 ml / min to 52.5 ml / min.

23. The epimedium extract according to claim 1, characterized in that In step (2), the relative density of the second thick paste at 55° C. to 70° C. is 1.08 to 1.

18.

24. The epimedium extract according to claim 1, characterized in that In step (3), the volume / volume ratio of the second thick paste to the water is 0.1-2.

25. The epimedium extract according to claim 24, characterized in that The volume / volume ratio of the second thick paste to the water is 0.95-1.

05.

26. The epimedium extract according to claim 1, characterized in that In step (3), the detection wavelength is 230-270 nm.

27. The epimedium extract according to claim 26, characterized in that The detection wavelength is 254 nm.

28. Use of the Epimedium extract according to any one of claims 1 to 27 in the preparation of a medicament for preventing and / or treating sarcopenia.

29. Use of a pharmaceutical composition in the preparation of a medicament for preventing and / or treating sarcopenia, characterized in that: The pharmaceutical composition consists of the epimedium extract according to any one of claims 1 to 27 and pharmaceutically acceptable excipients.

30. The use according to claim 29, characterized in that The auxiliary materials are selected from one or more of the following: diluents, excipients, wetting agents, binders, disintegrants, inclusion agents, flavoring agents, sustained-release agents, retention aids, lubricants, dispersants, plasticizers, opacifiers and antioxidants.

31. The use according to claim 29, characterized in that The dosage form of the pharmaceutical composition is powder, tablet, pill, capsule, film, lozenge, microcapsule, microsphere, granule, injection or oral solution.

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