Use of herba epimedii extract in prevention and treatment of sarcopenia

Abstract

A herba epimedii extract used for preventing and / or treating sarcopenia. A preparation method for the herba epimedii extract comprises the following steps: (1) weighing a proper amount of a herba epimedii medicinal material, adding water and performing decoction twice, filtering same, and performing concentration to obtain a first thick paste; (2) adding water to the first thick paste for dissolving same, performing standing treatment and centrifugation to obtain a supernatant, eluting the supernatant successively with a first solvent, a second solvent and a third solvent by means of a macroporous resin, collecting a third eluate, recovering the third solvent, and performing concentration to obtain a second thick paste; and (3) adding water to the second thick paste for diluting same, loading same into a preparative chromatographic column, performing elution by using a mobile phase, collecting a first fraction and / or a fifth fraction in different time periods, and performing concentration, drying and crushing, so as to obtain the herba epimedii extract. The herba epimedii extract has an effect on relieving sarcopenia.

Description

Application of epimedium extract in preventing and treating sarcopenia Technical Field

[0001] The present invention relates to the technical field of medicine, 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 a 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 the prevalence of sarcopenia in Chinese middle-aged and elderly people aged 60 to 70 years to be around 4.1% to 11.5%. The prevalence of sarcopenia in men and women aged 70 years and above was 13.2% and 4.8%, respectively. The incidence rate increased significantly in people over 80 years old, reaching as high as 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. 2019 Jan 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. Moreover, 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 (TCM) has accumulated considerable clinical experience in the treatment of sarcopenia. TCM classifies diseases characterized by thin, weak, and powerless muscles as "paralysis," which is similar to the clinical manifestations of sarcopenia, which are characterized by reduced muscle mass, decreased muscle strength, or decreased physical function. Because sarcopenia and paralysis have highly overlapping connotations, TCM often treats sarcopenia from the perspective of paralysis, with excellent results. Ancient TCM literature records that the onset of paralysis is closely related to the spleen and kidney, with the growth and aging of the human body being 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 of kidney qi. "Suwen: Treatise on Ancient Innocence" states: "At eight years old, a man's kidney qi is strong, his hair grows, and his teeth change. At twenty-eight, his kidney qi is abundant, the celestial essence arrives, and his essence and qi overflow, harmonizing yin and yang, enabling him to have children. At thirty-eight, his kidney qi is balanced, his tendons and bones are strong, and his true teeth grow and reach their full length. At forty-eight, his tendons and bones are thriving, and his muscles are full and strong. At fifty-eight, his kidney qi declines, and his hair falls and his teeth wither. At sixty-eight, his yang qi depletes from above, his face becomes scorched, and his hair and temples turn gray. At seventy-eight, his liver qi declines, and his tendons become unable to move. At eighty-eight, the celestial essence is exhausted, his essence is scarce, his kidneys decline, and his physical form reaches its limit, leading to the loss of hair and teeth." This indicates that the growth and aging of the human body, including the fullness and strength of muscles and their atrophy and shrinkage, changes with the rise and fall of kidney qi. As kidney qi declines, muscle function 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 its 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 men aged 80 years 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.

[0005] Summary of the Invention

[0006] Based on this, the present invention provides an epimedium extract for preventing and / or treating sarcopenia. The preparation method of the epimedium extract comprises the following steps:

[0007] (1) Weighing an appropriate amount of Epimedium brevicornum medicinal material, adding water to decoct, filtering to obtain a first filtrate and 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;

[0008] (2) dissolving the first thick paste in water, allowing it to stand, and 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, and eluting with a third solvent, collecting the third eluate, recovering the third solvent, and concentrating it to obtain a second thick paste; and

[0009] (3) diluting the second thick paste with water, loading the sample onto a preparative chromatography 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;

[0010] The first fraction is a fraction in the time period of 0 to 5 minutes;

[0011] The fifth fraction is a fraction within a time period of 20 to 40 minutes.

[0012] Furthermore, in step (1), the volume / mass (L / kg or ml / g) ratio of the water to the Epimedium medicinal material is 10-50, for example, 20-40.

[0013] Furthermore, the decocting time is 0.5 to 5 hours, for example 1 to 3 hours.

[0014] Furthermore, the concentration is concentration under reduced pressure.

[0015] Furthermore, the relative density of the first thick paste at 55° C. to 70° C. is 1.08 to 1.18.

[0016] Furthermore, in step (2), the first thick paste is cooled and then dissolved in water, wherein the cooling is cooling to room temperature.

[0017] 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.

[0018] Furthermore, the standing time is 1 to 10 hours, for example, 3 to 5 hours.

[0019] Furthermore, the macroporous resin is a D101 macroporous resin.

[0020] Furthermore, the volume / mass (L / kg or ml / g) ratio of the supernatant to the macroporous resin is 0.1-2, for example, about 0.4.

[0021] Furthermore, the first solvent is water, such as distilled water.

[0022] Furthermore, 4 to 10 times the volume of the column bed, for example, 6 to 8 times the volume of the column bed, are used for elution with the first solvent.

[0023] Furthermore, the flow rate of the distilled water is 20-80 ml / min, for example, about 50 ml / min.

[0024] Furthermore, the second solvent is an alcohol, such as ethanol.

[0025] Furthermore, the volume concentration of the second solvent is 10% to 30%, for example, about 20%.

[0026] 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.

[0027] Furthermore, the flow rate of the second solvent is 20-80 ml / min, for example, about 50 ml / min.

[0028] Furthermore, the third solvent is an alcohol, such as ethanol.

[0029] Furthermore, the volume concentration of the third solvent is 70% to 90%, for example, about 80%.

[0030] 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.

[0031] Furthermore, the flow rate of the third solvent is 20-80 ml / min, for example, about 50 ml / min.

[0032] Furthermore, the relative density of the second thick paste at 55° C. to 70° C. is 1.08 to 1.18.

[0033] Furthermore, in step (3), the volume / volume ratio of the second thick paste to the water is 0.1 to 2, for example, about 1.

[0034] Furthermore, the conditions of the preparative chromatography are: using a chromatographic column with octadecylsilane bonded silica gel as the filler, mobile phase A is selected from one or more of acetonitrile, methanol and tetrahydrofuran, 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.

[0035] Furthermore, the specifications of the chromatographic column are: column length 600 mm, inner diameter 250 mm, and particle size 10 μm.

[0036] Furthermore, phase A of the mobile phase is methanol.

[0037] Furthermore, the flow rate is 15-25 ml / min, for example, about 20 ml / min.

[0038] Furthermore, the detection wavelength is 230-270 nm, for example, 254 nm.

[0039] Furthermore, the column pressure is 15-25 MPa, for example, about 20 MPa.

[0040] 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.

[0041] 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.

[0042] 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.

[0043] Furthermore, the pharmaceutical composition further comprises one or more drugs and / or extracts for preventing and / or treating sarcopenia.

[0044] Furthermore, the drug is selected from one or more of the following: testosterone, selective androgen receptor modulators, growth hormone, ghrelin and myostatin inhibitors.

[0045] Furthermore, the pharmaceutical composition further comprises pharmaceutically acceptable excipients.

[0046] 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 aid, lubricant, dispersant, plasticizer, opacifier and antioxidant.

[0047] Furthermore, the dosage form of the pharmaceutical composition is powder, tablet, pill, capsule, film, lozenge, microcapsule, microsphere, granule, injection or oral solution.

[0048] 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 mass, reducing body weight, body fat and bone mineral content, and delaying the aging process.

[0049] Beneficial effects of the present invention:

[0050] 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, without a significant 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

[0051] In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the following briefly introduces the drawings required for use in the description of the embodiments. 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.

[0052] Figure 1 shows the effects of Epimedium Extract V on skeletal muscle in SAMP8 sarcopenia model mice. A shows HE staining of the tibialis anterior muscle; B shows DEXA results of body composition. DETAILED DESCRIPTION

[0053] The following will clearly and completely describe the technical solutions in the embodiments of the present invention in conjunction with the accompanying drawings. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative work shall fall within the scope of protection of the present invention.

[0054] Unless otherwise indicated, all technical and scientific terms and abbreviations used herein have the meanings commonly understood by those skilled in the art in the field of the present invention or the field in which the terms are used. Although any methods, conditions, substances or materials similar or equivalent to those disclosed herein can be used in the practice of the present invention, preferred methods, conditions, substances or materials are described herein.

[0055] The present invention is intended to encompass all alternatives, modifications, and equivalents that may come within the scope of 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 could be used in the practice of the present invention. The present invention is in no way limited to the methods and materials described.

[0056] 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.

[0057] In the present invention, the term "comprising" is synonymous with "including." As used herein, the terms "comprises," "including," "having," "containing," or any other variations thereof, are intended to cover a non-exclusive inclusion. For example, a composition, process, method, article, or apparatus that comprises a list of 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.

[0058] As described in the background technology section, there is currently 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 effectiveness of the epimedium extract claimed in the present invention in 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:

[0059] (1) Weighing an appropriate amount of Epimedium brevicornum medicinal material, adding water to decoct, filtering to obtain a first filtrate and 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;

[0060] (2) dissolving the first thick paste in water, allowing it to stand, and 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, and eluting with a third solvent, collecting the third eluate, recovering the third solvent, and concentrating it to obtain a second thick paste; and

[0061] (3) diluting the second thick paste with water, loading the sample onto a preparative chromatography 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;

[0062] The first fraction is a fraction in the time period of 0 to 5 minutes;

[0063] The fifth fraction is a fraction within a time period of 20 to 40 minutes.

[0064] 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.

[0065] In a preferred embodiment, the decocting time is 0.5 to 5 hours, such as 1 to 3 hours.

[0066] In a preferred embodiment, the concentration is concentration under reduced pressure.

[0067] In a preferred embodiment, the relative density of the first thick paste at 55° C. to 70° C. is 1.08 to 1.18.

[0068] 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.

[0069] 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, for example, about 1.

[0070] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 1" includes ±5% of 1, or from 0.95 to 1.05.

[0071] In a preferred embodiment, the standing time is 1 to 10 hours, such as 3 to 5 hours.

[0072] In a preferred embodiment, the macroporous resin is a D101 macroporous resin.

[0073] 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.

[0074] 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.

[0075] In a preferred embodiment, the first solvent is water, such as distilled water.

[0076] In a preferred embodiment, 4 to 10 times the column bed volume, such as 6 to 8 times the column bed volume, are used for elution with the first solvent.

[0077] In a preferred embodiment, the flow rate of the distilled water is 20-80 ml / min, for example, about 50 ml / min.

[0078] 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.

[0079] In a preferred embodiment, the second solvent is an alcohol, such as ethanol.

[0080] In a preferred embodiment, the volume concentration of the second solvent is 10% to 30%, for example, about 20%.

[0081] 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.

[0082] In a preferred embodiment, the second solvent is used for elution in an amount of 5 to 15 times the column bed volume, for example, 8 to 10 times the column bed volume.

[0083] In a preferred embodiment, the flow rate of the second solvent is 20-80 ml / min, for example, about 50 ml / min.

[0084] 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.

[0085] In a preferred embodiment, the third solvent is an alcohol, such as ethanol.

[0086] In a preferred embodiment, the volume concentration of the third solvent is 70% to 90%, for example, about 80%.

[0087] 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.

[0088] In a preferred embodiment, the third solvent is used for elution in an amount of 5 to 15 times the column bed volume, for example, 8 to 10 times the column bed volume.

[0089] In a preferred embodiment, the flow rate of the third solvent is 20-80 ml / min, for example, about 50 ml / min.

[0090] 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.

[0091] In a preferred embodiment, the relative density of the second thick paste at 55° C. to 70° C. is 1.08 to 1.18.

[0092] 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.

[0093] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 1" includes ±5% of 1, or from 0.95 to 1.05.

[0094] In a preferred embodiment, the preparative chromatography conditions are: using a chromatographic column filled with 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.

[0095] In a preferred embodiment, the specifications of the chromatographic column are: column length 600 mm, inner diameter 250 mm, and particle size 10 μm.

[0096] In a preferred embodiment, phase A of the mobile phase is methanol.

[0097] In a preferred embodiment, the flow rate is 15-25 ml / min, such as about 20 ml / min.

[0098] 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.

[0099] In a preferred embodiment, the detection wavelength is 230-270 nm, for example, 254 nm.

[0100] In a preferred embodiment, the column pressure is 15-25 MPa, such as about 20 MPa.

[0101] 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.

[0102] 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.

[0103] 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.

[0104] 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.

[0105] In a preferred embodiment, the pharmaceutical composition further comprises one or more drugs and / or extracts for preventing and / or treating sarcopenia.

[0106] 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.

[0107] In a preferred embodiment, the pharmaceutical composition further comprises a pharmaceutically acceptable excipient.

[0108] 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.

[0109] 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 a subject.

[0110] In the present invention, the term "pharmaceutically acceptable excipient" refers to carriers and / or excipients that are pharmacologically and / or physiologically compatible with the subject and the active ingredient (i.e., capable of eliciting 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).

[0111] In a preferred embodiment, the excipient is selected from one or more of the following: diluents, excipients, excipients, wetting agents, binders, disintegrants, inclusion agents, flavoring agents, sustained-release agents, retention aids, lubricants, dispersants, plasticizers, opacifiers and antioxidants.

[0112] Those skilled in the art will know how to select the specific chemical substance within the scope of above-mentioned auxiliary material classification.For example, this diluent can be selected from following one or more: Icing Sugar, starch, compressible starch, lactose, dextrin, mannitol, sorbitol, microcrystalline cellulose, calcium sulfate and calcium carbonate.This wetting agent can be selected from following one or more: stearic acid polyoxymethylene, poloxamer, polyoxyethylene sorbitan fatty acid ester, polyoxyethylene hydrogenated castor oil, polyoxyethylene alkyl ether, polysorbate such as polysorbate 80, spermol, glycerol fatty acid ester (such as triacetin, glyceryl monostearate and analogue), 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.This adhesive can be selected from following one or more: 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 pregelatinized corn starch. The plasticizer may be dibutyl sebacate and / or various citrate esters. The sustained-release agent may be selected from one or more of the following: sodium carboxymethylcellulose, low-substituted hydroxypropyl cellulose, hydroxypropyl methylcellulose, microcrystalline cellulose, hydroxyethyl cellulose, gum arabic, gelatin, and shellac. The antioxidant may be selected from one or more of the following: sodium bisulfite, sodium metabisulfite, 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.

[0113] These excipients are preferably pharmaceutically inert, or may have a synergistic or additive effect to enhance the therapeutic activity of the pharmaceutical composition. The above excipients are merely enumerated. The excipients actually used in the present invention are not limited to the above excipients and may be adjusted according to actual conditions to achieve the effects of the present invention.

[0114] 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.

[0115] 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 mass, reducing body weight, body fat and bone mineral content, and delaying the aging process.

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

[0117] According to another aspect of the present invention, provided is a method for preventing and / or treating sarcopenia in a subject, comprising administering to the subject an effective amount of the above-mentioned Epimedium extract or the above-mentioned pharmaceutical composition.

[0118] In the present invention, the term "subject" refers to a mammal. The mammal may be a human, non-human primate, mouse, rat, dog, cat, horse, or 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.

[0119] 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 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., several weeks to several months) or a longer period of time (several months to several years).

[0120] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are intended to illustrate the present invention only and are not intended to limit the scope of the invention. The experimental methods in the following examples, for which specific conditions are not specified, are generally based on conventional conditions or conditions recommended by the manufacturer.

[0121] 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 in any way. 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.

[0122] Example

[0123] Preparation Example

[0124] 1. Experimental Materials

[0125] 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 grade and were purchased from Thermo Fisher Scientific. Trizol reagent (lot number: G826KA6730), DEPC water, and primers were purchased from Shanghai Sangon Biotechnology Co., Ltd. Chloroform, formaldehyde, anhydrous ethanol, and sodium carboxymethylcellulose (CMC-Na) were purchased from Sinopharm Chemical Reagent Co., Ltd. Isopropanol was purchased from Merck, Germany. An RNA reverse transcription kit was purchased from Applied Biological Materials, Canada. The Green Pro Taq HS premixed qPCR kit (including ROX) was purchased from Hunan Aikerui Bioengineering Co., Ltd. TNF-α and IFN-γ were purchased from Shanghai Biyuntian Biotechnology Co., Ltd., and Epimedium herb was purchased from Shanghai Kangqiao Decoction Pieces Factory, conforming to the 2020 edition of the Chinese Pharmacopoeia. Ultrapure water was used, and all other reagents were of analytical grade.

[0126] 2. Experimental methods

[0127] Take the herbal medicine of Epimedium, add 20 to 40 times the volume of the herbal medicine in water and boil 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 boil for 1 to 3 hours, filter and set aside the filtrate; combine the filtrates obtained from the two extractions, concentrate under reduced pressure to a thick paste 1 with a relative density d = 1.08 to 1.18 (55 to 70 ° C), let it cool (to room temperature), add water of equal weight to the herbal medicine 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 50 ml / 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 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%.

[0128] The fractions were collected at the following time periods:

[0129] Fractions F5-8 and F9-12 were combined to obtain a total of 6 fractions, which were concentrated, dried, and pulverized to obtain 6 extract powders, which were named extracts I (F1), II (F2), III (F3), IV (F4), V (F5-8), and VI (F9-12).

[0130] Pharmacodynamic Examples

[0131] Experiment 1: Effects of Epimedium Extract on Myoblasts

[0132] 1. Experimental Purpose

[0133] The effects of different extracts of Epimedium on the myogenic differentiation of C2C12 cells were evaluated by examining the effects of drugs on myoblast differentiation.

[0134] 2. Experimental methods

[0135] 2.1 C2C12 cell culture and induction of myogenic differentiation

[0136] When the C2C12 myoblasts grew to 70% to 80% fusion, 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 myogenic differentiation for 6 days to form myotubes. The differentiation medium was replaced with new one-day medium and culture was continued.

[0137] 2.2 Cell grouping and drug administration

[0138] C2C12 myoblasts were randomly divided into a control group, a TNF-α / IFN-γ stimulation group, or an epimedium extract I-VI (10 and 50 μM) group, with triplicate wells per group. The control group was cultured in growth medium for 24 hours; the stimulation group was cultured in growth medium containing 10 ng / mL TNF-α and 10 ng / mL IFN-γ for 24 hours; and the epimedium extract I-VI groups were cultured in growth medium containing 10 ng / mL TNF-α, 10 ng / mL IFN-γ, and epimedium extracts I-VI (10 and 50 μM) for 24 hours. On day 5 of myogenic differentiation induction, each group was cultured in differentiation medium containing or without the drug for 24 hours. Myotube morphology was observed and photographed in real time under an inverted fluorescence microscope.

[0139] 2.3 CCK8 assay for cell viability

[0140] C2C12 myoblasts in logarithmic growth phase were cultured at 5×10 3 Cells were seeded in 96-well plates at a density of 100 μM and cultured overnight until adhered to the wall. Different concentrations of Epimedium extracts I to 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 a microplate reader to calculate the cell viability. Three replicate wells were set up for each group.

[0141] 2.4 Detection of mRNA expression of MRFs family factors in myotubes differentiated from myoblasts by RT-qPCR

[0142] Cells were treated as described in "2.2." Myoblasts and differentiated myotubes were collected and lysed with RZ lysis buffer. RNA was extracted using a total RNA extraction kit and reverse transcribed to generate cDNA. mRNA expression of inflammatory factors in myoblasts and MRF family factors in differentiated myotubes was assayed. Analysis was performed using the 2-ΔΔCT method, using GAPDH as an internal control. Primer sequences were obtained from PrimerBank (Table 1) and synthesized by Sangon Biotech (Shanghai) Co., Ltd.

[0143] Table 1 Primer sequences

[0144] 3. Experimental results

[0145] 3.1 Effects of different epimedium extracts on C2C12 myoblast viability

[0146] 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.

[0147] Table 2 Effects of different extracts of Epimedium on the viability of C2C12 myoblasts

[0148] 3.2 Effects of different epimedium extracts on differentiated myotube cell morphology and MRFs gene expression

[0149] The results are shown in Table 3. Compared with the control group, the myotube nuclear fusion in the stimulation group was reduced, the morphology was atrophied, and the diameter and length were reduced. The myotube nuclear fusion in the 50 μM epimedium extract I and V group 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.

[0150] 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.

[0151] Table 3 Effects of different extracts of Epimedium on the expression of MyoD, Mrf5, Mygenin and Myf6 mRNA in C2C12 myoblasts Note: * indicates P < 0.05 compared with the stimulation group; ** indicates P < 0.01 compared with the stimulation group; *** indicates P < 0.001 compared with the stimulation group;

[0152] 4. Conclusion

[0153] The results of cell experiments showed that Epimedium Extract V could significantly promote myoblast differentiation, and had the best effect.

[0154] Experiment 2: Effects of Epimedium Extract V on SAMP8 Sarcopenia Model Mice

[0155] 1. Experimental Materials

[0156] 1.1 Experimental animals

[0157] 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.

[0158] The experimental animals were housed in an SPF-grade animal room at the Shanghai Laboratory Animal Center. The temperature was (24 ± 1)°C, the relative humidity was 50%–70%, and the light / dark cycle was 12 h / 12 ​​h. The experimental procedures strictly adhered to the relevant regulations and provisions of the Ethics Committee of the Shanghai Laboratory Animal Center.

[0159] 1.2 Experimental drugs

[0160] The Epimedium extract V used in the embodiment of the present invention is homemade in the laboratory.

[0161] 1.3 Experimental instruments

[0162] Dual-energy X-ray absorptiometry (iNSiGHT VET DXA, OsteoSys, Korea); YLS-13A muscle strength tester, Jinan Yiyan Technology Co., Ltd.

[0163] 2 Methods

[0164] 2.1 Grouping and Dosing

[0165] After adaptive feeding with ordinary feed for one week, 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.

[0166] Epimedium Extract V was dissolved in 0.3% sodium carboxymethylcellulose solution. Each treatment group received different concentrations of Epimedium Extract V solution by gavage once daily for 12 weeks, with a gavage volume of 0.1 mL / 10 g. The negative control group and the model group received 0.3% sodium methylcellulose solution by gavage once daily for 12 weeks, with a gavage volume of 0.1 mL / 10 g.

[0167] 2.2 Detection indicators and methods

[0168] 2.2.1 General determination

[0169] The body weight and food intake of mice were measured according to conventional methods.

[0170] Method for measuring mouse body weight and food intake: Mouse weight was measured according to standard weighing procedures. Experimental equipment: electronic balance (range: 0-200g, sensitivity 0.01g). Operating steps: Select a suitable height and stable position to place the electronic scale; plug in the power supply, turn on the electronic balance, and after it is ready, place an appropriately sized weighing box on it and adjust it to zero. According to the "Operational Procedures for Grasping and Fixing Laboratory Animals", grab the mouse from the feeding box, place it in the weighing box, and read the value after the value stabilizes. If the value fluctuates continuously, select the middle value and record it in a dedicated experimental original record sheet. Place the mouse back in the cage, cover the cage, return it to the cage rack, and insert the water bottle. After weighing, put the mouse cage in order, turn off the balance, disconnect the power supply, wipe the electronic balance clean, and return it to its original position. Fill in the electronic balance usage record.

[0171] The food intake of mice was measured using a 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 housed 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. Data such as food intake (grams) can be monitored through the data dashboard. Mouse feed was purchased from the Shanghai Laboratory Animal Research Center and produced according to the standardized quality requirements of laboratory animal compound feed.

[0172] 2.2.2 Aging score

[0173] The degree of apparent aging was assessed using the Aging Rating Scale, which includes 11 categories, including reactive, passive, skin and hair, eye, and spine-related aging. Each category is scored from 0 to 4 points, and assessments were conducted every 0, 4, 8, and 12 weeks.

[0174] 2.2.3 Mouse muscle strength measurement

[0175] 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. The mice were then gently pulled back by their tails, with their posture parallel to the table surface, until they released the mesh. The peak force exerted by the mouse's limbs was recorded in grams (g). Evaluations were performed every 0, 4, 8, and 12 weeks, with each mouse tested three times, the average value was taken, and the weight was used for statistical analysis.

[0176] 2.2.4 Body composition determination of mice

[0177] Three days before the mice were sacrificed, dual-energy X-ray absorptiometry (DEXA) was used to measure body composition, including body fat content, muscle content, and bone mineral content. The data were analyzed using the software provided by the manufacturer.

[0178] 2.2.5 Skeletal muscle pathology detection

[0179] After the intervention, mice were killed and the tibialis anterior muscle was harvested. The tibialis anterior muscle was fixed with 4% paraformaldehyde for 24 hours, dehydrated, embedded in paraffin, and cut into 5 μm paraffin sections. Skeletal muscle sections were stained with hematoxylin and eosin to examine the morphology and number of skeletal muscle fibers.

[0180] 2.2.6 Statistical methods

[0181] 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 between groups. P < 0.05 was considered statistically significant.

[0182] 3 Results

[0183] 3.1 Effect of Epimedium Extract V on Food Intake in SAMP8 Sarcopenia Model Mice

[0184] 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; eight weeks after administration, the food intake of the model group and the negative control group increased compared with that of the four weeks after administration, while 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, the food intake of the low-dose group increased compared with the model group (P < 0.05), indicating that Epimedium Extract V had no significant effect on the food intake of SAMP8 sarcopenia model mice.

[0185] Table 4 Effects of Epimedium Extract V on food intake in SAMP8 sarcopenia model mice

[0186] 3.2 Effect of Epimedium Extract V on Body Weight in SAMP8 Sarcopenia Model Mice

[0187] The results are shown in Table 5. There was no significant difference in body weight between the groups before administration, indicating that the groups were comparable. There was no significant difference in the body weight of mice between 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, while the body weight of mice in the other groups decreased compared with before. There was no significant difference in the body weight of mice between the groups 8 weeks after administration, and the body weight of mice was basically the same as that 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), suggesting that Epimedium Extract V may have a certain weight loss effect on the body weight of SAMP8 sarcopenia model mice.

[0188] Table 5 Effects of Epimedium Extract V on the body weight of SAMP8 sarcopenia model mice Note: Compared with the model group, *P<0.05, **P<0.01.

[0189] 3.3 Effect of Epimedium Extract V on Aging in SAMP8 Sarcopenia Model Mice

[0190] The results are shown in Table 6. Before administration, there were no significant differences in the aging scores of SAMP8 mice between the groups, indicating that the groups were comparable. After 4 weeks of administration, compared with the model group, the aging scores of the medium-dose and high-dose groups were 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 and high-dose groups were reduced (P < 0.05). After 8 weeks of administration, compared with the model group, the aging scores of the medium-dose and high-dose groups were 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 score of the medium-dose group was reduced (P < 0.05), and the aging score of the high-dose group showed a downward 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 showed a downward trend.

[0191] Table 6 Effects of Epimedium Extract V on the aging degree of SAMP8 sarcopenia model mice Note: Compared with the model group, *P<0.05, **P<0.01; compared with the low-dose group, △ P<0.05, △△ P<0.01.

[0192] 3.4 Effect of Epimedium Extract V on Muscle Strength in SAMP8 Sarcopenia Model Mice

[0193] The results are shown in Table 7. Before administration, muscle strength in all groups was significantly decreased compared with the negative control group (P < 0.01). There were no significant differences in muscle strength among the SAMP8 mice, indicating that the experimental system was reliable and the groups were comparable. After 4 weeks of administration, muscle strength in the medium-dose, high-dose, and negative control groups was significantly increased compared with the model group (P < 0.01); compared with the low-dose group, muscle strength in the medium-dose and high-dose groups was increased (P < 0.05). After 8 weeks of administration, muscle strength in the medium-dose, high-dose, and negative control groups was significantly increased (P < 0.01). After 12 weeks of administration, muscle strength in the high-dose group was increased compared with the model group (P < 0.05), and muscle strength in the medium-dose and negative control groups was significantly increased (P < 0.01). Compared with the low-dose group, muscle strength in the medium-dose group was increased (P < 0.05), and muscle strength in the high-dose group showed a trend toward improvement.

[0194] Table 7 Effects of Epimedium Extract V on Muscle Strength in SAMP8 Sarcopenia Model Mice Note: Compared with the model group, *P<0.05, **P<0.01; compared with the low-dose group, △ P<0.05,△△ P<0.01.

[0195] 3.5 Effects of Epimedium Extract V on Body Fat, Muscle Mass, and Bone Mineral Content (BMC) in SAMP8 Sarcopenia Model Mice

[0196] The results after 12 weeks of administration are shown in Table 8. Compared with the model group, the medium-dose group showed a significant decrease in body fat content, a significant increase in muscle content (P < 0.01), and a decrease in BMC (P < 0.05). In the negative control group, body fat content decreased, muscle content increased (P < 0.05), and BMC showed a decreasing trend. In the high-dose group, BMC and body fat content showed a decreasing trend, while muscle content showed an increasing trend. DEXA images of the body composition of SAMP8 mice are shown in Figure 1B.

[0197] Table 8 Effects of Epimedium Extract V on body fat content, muscle content and bone mineral content in SAMP8 sarcopenia model mice Note: Compared with the model group, *P<0.05, **P<0.01.

[0198] 3.6 Effects of Epimedium Extract V on Skeletal Muscle Sections of SAMP8 Sarcopenia Model Mice

[0199] HE-stained sections were used to analyze the pathological damage of skeletal muscle in 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 helps to restore the pathological damage of skeletal muscle in aging mice, and the performance of the medium-dose group is particularly prominent. Sections of the tibialis anterior muscle of SAMP8 mice are shown in Figure 1A.

[0200] 4 Summary

[0201] Epimedium extract V can increase muscle strength and muscle content in SAMP8 sarcopenia model mice, reduce body weight, body fat and BMC content, and delay the aging process, without significantly affecting food intake, indicating that epimedium extract V can be used as a drug for the treatment of sarcopenia.

[0202] 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 intended to help understand the method of the present invention and its core concept. At the same time, changes or modifications made by those skilled in the art based on the concept of the present invention, the specific implementation methods of the present invention, and the scope of application are all within the scope of protection of the present invention. In summary, the contents 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, allowing it to stand, centrifuging it to obtain a supernatant, passing the supernatant through a macroporous resin, eluting it with a first solvent, discarding the first eluent, eluting it with a second solvent, discarding the second eluent, eluting it with a third solvent, collecting the third eluent, recovering the third solvent and concentrating it to obtain a second thick paste; as well as (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; 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 (L / kg or ml / g) ratio of the water to the Epimedium medicinal material is 10 to 50, for example, 20 to 40; Preferably, the decoction time is 0.5 to 5 hours, for example 1 to 3 hours; More preferably, the concentration is reduced pressure concentration; Also preferably, the relative density of the first thick paste at 55° C. to 70° C. is 1.08 to 1.

18.

3. 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; Preferably, the mass / mass (kg / kg or g / g) ratio of the water to the Epimedium medicinal material is 0.1 to 2, for example, about 1; Preferably, the standing time is 1 to 10 hours, for example 3 to 5 hours; More preferably, the macroporous resin is a D101 macroporous resin; More preferably, the volume / mass (L / kg or ml / g) ratio of the supernatant to the macroporous resin is 0.1 to 2, such as about 0.4; Also preferably, the first solvent is water, such as distilled water; Preferably, 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; Preferably, the flow rate of the distilled water is 20 to 80 ml / min, for example, about 50 ml / min; Still more preferably, the second solvent is an alcohol, such as ethanol; More preferably, the volume percentage concentration of the second solvent is 10% to 30%, for example, about 20%; More preferably, 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; Still more preferably, the flow rate of the second solvent is 20 to 80 ml / min, for example about 50 ml / min; Particularly preferably, the third solvent is an alcohol, such as ethanol; Particularly preferably, the volume concentration percentage of the third solvent is 70% to 90%, for example, about 80%; Particularly preferably, 5 to 15 times the volume of the column bed, for example 8 to 10 times the volume of the column bed, are used for elution with the third solvent; Particularly preferably, the flow rate of the third solvent is 20 to 80 ml / min, for example, about 50 ml / min; Particularly preferably, the relative density of the second thick paste at 55° C. to 70° C. is 1.08 to 1.

18.

4. 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 to 2, for example, about 1; Preferably, 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: 0-20min, 25% A; 20-40min, 40% A; 40-60min, 48% A, the flow rate is 10-30ml / min, the detection wavelength is 150-350nm, and the column pressure is 10-30Mpa; More preferably, the specifications of the chromatographic column are: column length 600 mm, inner diameter 250 mm, particle size 10 μm; Preferably, the mobile phase A is methanol; Still more preferably, the flow rate is 15 to 25 ml / min, for example about 20 ml / min; Still more preferably, the detection wavelength is 230-270 nm, for example 254 nm; Even more preferably, the column pressure is 15-25 MPa, such as about 20 MPa.

5. Use of the epimedium extract according to any one of claims 1 to 4 in the preparation of a medicament for preventing and / or treating sarcopenia.

6. Use of a pharmaceutical composition comprising the Epimedium extract according to any one of claims 1 to 4 in the preparation of a medicament for preventing and / or treating sarcopenia.

7. The use according to claim 6, characterized in that The pharmaceutical composition further comprises one or more drugs and / or extracts for preventing and / or treating sarcopenia; Preferably, the drug is selected from one or more of the following: testosterone, selective androgen receptor modulators, growth hormone, ghrelin and myostatin inhibitors; Also preferably, the pharmaceutical composition further comprises a pharmaceutically acceptable excipient.

8. The use according to claim 7, 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.

9. The use according to claim 7, characterized in that: The dosage form of the pharmaceutical composition is powder, tablet, pill, capsule, film, lozenge, microcapsule, microsphere, granule, injection or oral solution.

10. The use according to any one of claims 5 to 9, characterized in that The prevention and / or treatment of sarcopenia is manifested in 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.

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