Use of mulberry extract in preparation of drug for treating and / or preventing male infertility

Abstract

Use of a mulberry extract in the preparation of a drug for treating and / or preventing male infertility. The extract comprises an alkaloid with a weight content of 3% or more, and / or a polysaccharide with a weight content of not more than 70%, and / or flavone with a weight content of not more than 10%, and / or an amino acid with a weight content of not more than 50%, and / or other components. The extract can be used for male infertility related to male sexual dysfunction and male reproductive dysfunction.

Description

Use of mulberry extract in preparing medicine for treating and / or preventing male infertility Technical Field

[0001] The present invention relates to the field of biomedicine, and in particular to use of a mulberry extract in preparing a medicine for treating and / or preventing male infertility. Technical Background

[0002] Male infertility is defined as male-factor-related infertility when couples of childbearing age fail to conceive naturally after having normal sexual intercourse for more than one year without taking contraceptive measures. It is estimated that 15% of couples of childbearing age worldwide experience fertility problems, with male-factor-related factors accounting for approximately 50%. Male infertility is not a single disease; it is often the result of multiple causes and factors. Key risk factors include age, unhealthy habits (smoking, drinking), obesity, unhealthy lifestyles (staying up late, lack of exercise), pathogen infection, varicocele, and adverse psychological factors.

[0003] Clinically, male infertility is categorized into two types: sexual dysfunction and normal sexual function. The latter is often referred to as "reproductive dysfunction." Clinically, sexual dysfunction and reproductive dysfunction interact and reinforce each other, often not existing independently. Sexual dysfunction often manifests as sexual desire disorder, erectile dysfunction, ejaculation disorder, and anorgasmia: sexual desire disorder is primarily associated with decreased testosterone levels or altered testosterone receptor function; erectile dysfunction is primarily related to damage to the corpus cavernosum smooth muscle or endothelial cell dysfunction; ejaculation disorder is primarily related to damage to smooth muscle and autonomic nerves, as well as decreased testosterone levels; and anorgasmia is primarily associated with androgen and pudendal sensory nerve disorders. Erectile dysfunction and ejaculation disorder severely affect the fertilization of sperm and fertilized egg, and are closely related to male infertility. Reproductive dysfunction is mostly caused by abnormal sperm parameters, such as decreased sperm count and poor sperm motility. Clinical manifestations are mostly manifested as vas deferens obstruction, abnormal sperm quality, semen abnormalities, varicocele and inflammation. Among them, sperm quality is an important indicator for evaluating male infertility. According to the results of semen analysis, it can be further divided into azoospermia, oligospermia, asthenozoospermia, asthenozoospermia and infertility with normal sperm count.

[0004] Male reproductive and sexual dysfunction seriously impacts people's quality of life and fertility, but existing treatments often fall short of expectations and have poor clinical efficacy. Especially in the face of a severely aging population and the national policy of promoting two and three children, developing safe and effective medications that can both improve sperm quality and motility and alleviate sexual dysfunction is not only of great scientific and medical significance, but also of significant social value. Summary of the Invention

[0005] The present invention aims to provide a use of a mulberry extract in the preparation of a medicament for treating and / or preventing male infertility; on the other hand, to provide a use of a mulberry extract in the treatment and / or prevention of male infertility; on the other hand, to provide a method for treating male infertility, comprising administering a therapeutically or preventively effective amount of a mulberry extract to a patient in need thereof; the mulberry extract has a significant therapeutic effect on male infertility including those associated with sexual dysfunction and reproductive dysfunction.

[0006] On the other hand, the present invention provides the use of mulberry extract in preparing a product for inhibiting high glucose-induced apoptosis of Leydig cells in vitro; on the other hand, the present invention provides the use of mulberry extract in inhibiting high glucose-induced apoptosis of Leydig cells in vitro; on the other hand, the present invention provides a method for inhibiting high glucose-induced apoptosis of Leydig cells in vitro, comprising administering a therapeutically or preventively effective amount of mulberry extract to a patient in need thereof.

[0007] On the other hand, the present invention provides the use of mulberry extract in preparing a product for improving high glucose-induced endothelial cell damage in vitro; on the other hand, the present invention provides the use of mulberry extract in improving high glucose-induced endothelial cell damage in vitro; on the other hand, the present invention provides a method for improving high glucose-induced endothelial cell damage in vitro, comprising administering a therapeutically or preventively effective amount of mulberry extract to a patient in need thereof.

[0008] In the present invention, based on the weight percentage of the sum of the components of the mulberry extract as 100%, the mulberry extract contains 3% or more of alkaloids by weight (optionally containing 3-99% of alkaloids by weight, further optionally containing 15-99% of alkaloids by weight, further optionally containing 30-99% of alkaloids by weight, further optionally containing 40-99% of alkaloids by weight, further optionally containing 50-99% of alkaloids by weight, further optionally containing 60-99% of alkaloids by weight, further optionally containing 35-70% of alkaloids by weight, further optionally containing 60-75% of alkaloids by weight);

[0009] and / or contains no more than 70% polysaccharide by weight (optionally containing 0.2-70% polysaccharide by weight, further optionally containing 0.2-50% polysaccharide by weight, further optionally containing 0.2-35% polysaccharide by weight, further optionally containing 0.2-25% polysaccharide by weight, further optionally containing 0.2-23% polysaccharide by weight, further optionally containing 20-25% polysaccharide by weight),

[0010] and / or contains flavonoids in an amount not higher than 10% by weight (optionally containing flavonoids in an amount of 0.05-5% by weight, further optionally containing flavonoids in an amount of 0-2% by weight, further optionally containing flavonoids in an amount of 0.05-2% by weight, further optionally containing flavonoids in an amount of 0.5-1.5% by weight, further optionally containing flavonoids in an amount of 0-1% by weight, further optionally containing flavonoids in an amount of 0.05-1% by weight),

[0011] and / or contains no more than 50% by weight of amino acids (optionally containing 0-30% by weight of amino acids, further optionally containing 0-25% by weight of amino acids, further optionally containing 0-20% by weight of amino acids, further optionally containing 0-5% by weight of amino acids, further optionally containing 3-25% by weight of amino acids or further optionally containing 5-20% by weight of amino acids),

[0012] and / or other components (the weight content is optionally 0-25%, further optionally 0-20%, further optionally 0-15%, further optionally 0-11%, further optionally 2-20%, further optionally 4-8% by weight.

[0013] Preferably, male infertility is male sexual dysfunction and / or male reproductive dysfunction.

[0014] Preferably, male sexual dysfunction and / or male reproductive dysfunction is caused by endocrine abnormalities and / or metabolic abnormalities; preferably, the metabolic abnormality is abnormal glucose and lipid metabolism; preferably, the endocrine abnormality is abnormal hormone levels.

[0015] Preferably, the abnormal glucose and lipid metabolism is selected from at least one of diabetes, hyperglycemia, overweight, obesity, and hyperlipidemia; preferably, the abnormal hormone level is abnormal sex hormone level, preferably abnormal androgen level.

[0016] Preferably, the male sexual dysfunction is selected from at least one of sexual desire disorder, erectile dysfunction, ejaculation disorder, and anorgasmia; preferably, the male sexual dysfunction is erectile dysfunction and / or ejaculation disorder.

[0017] Preferably, the male reproductive dysfunction is selected from at least one of vas deferens obstruction, sperm quality abnormalities, semen abnormalities, varicocele, and inflammation; further preferably, the sperm quality abnormalities are selected from at least one of azoospermia, oligospermia, asthenospermia, and asthenospermia. Preferably, the male reproductive dysfunction is oligospermia, asthenospermia, and / or oligoasthenospermia.

[0018] Preferably, the treatment and / or prevention of male infertility is specifically embodied in at least one of the following:

[0019] 1) Promote spermatogenesis;

[0020] 2) Improve sperm motility;

[0021] 3) Improve sperm quality;

[0022] 4) Improve testicular index;

[0023] 5) Improve the structure of testicular seminiferous tubules and the morphology and number of spermatogenic cells;

[0024] 6) Improving relevant hormone levels; preferably, the sex hormone is androgen; further preferably, the androgen is testosterone (T);

[0025] 7) Improving the expression level of androgen synthase; the androgen synthase includes at least one of the testosterone synthesis rate-limiting enzyme steroidogenic acute regulatory protein (STAR), the testosterone synthesis important enzyme cytochrome P450 17α-hydroxylase (CYP17A1), and 17β-hydroxysteroid dehydrogenase (17β-HSD);

[0026] 8) Improve androgen receptor expression levels;

[0027] 9) Improving penile tissue damage and / or cell damage; preferably, the penile cells are penile endothelial cells; preferably, improving the NO content in the penile endothelial cells; preferably, improving penile tissue damage specifically includes improving at least one of the average thickness of the spongy scaffold, the connection density, the separation degree, and the average intercept length of the sinusoidal space.

[0028] Further preferably, the treatment and / or prevention of male reproductive dysfunction is specifically embodied in at least one of the following:

[0029] 1) Promote spermatogenesis;

[0030] 2) Improve sperm motility;

[0031] 3) Improve sperm quality;

[0032] 4) Improve testicular index;

[0033] 5) improving the structure of the seminiferous tubules of testicular tissue and the morphology and number of spermatogenic cells; preferably, the spermatogenic cells include at least one of spermatogonia, primary spermatocytes, secondary spermatocytes and spermatids;

[0034] 6) Improving relevant hormone levels; preferably, the sex hormone is androgen; further preferably, the androgen is testosterone (T);

[0035] 7) Improving the expression level of androgen synthase; preferably, the androgen synthase includes at least one of the testosterone synthesis rate-limiting enzyme steroidogenic acute phase regulatory protein STAR, the testosterone synthesis important enzyme cytochrome P45017α-hydroxylase CYP17A1, and 17β-hydroxysteroid dehydrogenase 17β-HSD

[0036] 8) Improve androgen receptor expression levels;

[0037] 9) Improving testicular cell damage; preferably, the testicular cells are Leydig cells;

[0038] Further preferably, the treatment and / or prevention of male sexual dysfunction is specifically embodied in at least one of the following:

[0039] 1) Improving the level of relevant hormones; preferably, the sex hormone is androgen; further preferably, the androgen is testosterone (T);

[0040] 2) Improving the expression level of androgen synthase; preferably, the androgen synthase includes at least one of the testosterone synthesis rate-limiting enzyme steroidogenic acute phase regulatory protein STAR, the testosterone synthesis important enzyme cytochrome P45017α-hydroxylase CYP17A1, and 17β-hydroxysteroid dehydrogenase 17β-HSD;

[0041] 3) Improve androgen receptor expression levels;

[0042] 4) improving penile tissue and / or cell damage; preferably, the penile cells are penile endothelial cells; preferably, improving the NO content in the penile endothelial cells; preferably, improving penile tissue damage includes improving at least one of the average thickness of the spongy scaffold, the connection density, the separation degree, and the average intercept length of the sinusoidal space;

[0043] 5) Improvement of IIEF-15 score; preferably, the IIEF-15 score includes at least one of erectile function, satisfaction with penetration, orgasm, sexual desire and overall satisfaction.

[0044] In an optional embodiment of the present invention, taking the sum of the weight percentages of the components of the mulberry extract as 100%, the weight contents of the components in the mulberry extract are:

[0045] Optionally, taking the sum of the weight percentages of the components of the mulberry extract as 100%, the weight percentages of the components in the mulberry extract are:

[0046] Optionally, taking the sum of the weight percentages of the components of the mulberry extract as 100%, the weight percentages of the components in the mulberry extract are:

[0047] Optionally, taking the sum of the weight percentages of the components of the mulberry extract as 100%, the weight contents of the components in the mulberry extract are:

[0048] Optionally, taking the sum of the weight percentages of the components of the mulberry extract as 100%, the weight contents of the components in the mulberry extract are:

[0049] Optionally, taking the sum of the weight percentages of the components of the mulberry extract as 100%, the weight percentages of the components in the mulberry extract are:

[0050] Optionally, taking the sum of the weight percentages of the components of the mulberry extract as 100%, the weight percentages of the components in the mulberry extract are:

[0051] Further optionally, taking the sum of the weight percentages of the components of the mulberry extract as 100%, the weight contents of the components in the mulberry extract are:

[0052] Further optionally, taking the sum of the weight percentages of the components of the mulberry extract as 100%, the weight contents of the components in the mulberry extract are:

[0053] Further optionally, taking the sum of the weight percentages of the components of the mulberry extract as 100%, the weight contents of the components in the mulberry extract are:

[0054] More optionally, based on the mulberry extract, the weight content of each component is:

[0055] Optionally, based on the mulberry extract, the weight content of each component is:

[0056] Optionally, based on the mulberry extract, the weight content of each component is:

[0057] Optionally, based on the mulberry extract, the weight content of each component is:

[0058] Optionally, based on the mulberry extract, the weight content of each component is:

[0059] Optionally, based on the mulberry extract, the weight content of each component is:

[0060] Optionally, based on the mulberry extract, the weight content of each component is:

[0061] Optionally, based on the mulberry extract, the weight content of each component is:

[0062] Optionally, based on the mulberry extract, the weight content of each component is:

[0063] Optionally, based on the mulberry extract, the weight content of each component is:

[0064] Optionally, based on the mulberry extract, the weight content of each component is:

[0065] Optionally, based on the mulberry extract, the weight content of each component is:

[0066] Optionally, based on the mulberry extract, the weight content of each component is:

[0067] Optionally, based on the mulberry extract, the weight content of each component is:

[0068] Optionally, based on the mulberry extract, the weight content of each component is:

[0069] Optionally, based on the mulberry extract, the weight content of each component is:

[0070] Optionally, based on the mulberry extract, the weight content of each component is:

[0071] Optionally, based on the mulberry extract, the weight content of each component is:

[0072] Optionally, based on the mulberry extract, the weight content of each component is:

[0073] The weight content of each component in the mulberry extract is:

[0074] The weight content of each component in the mulberry extract is:

[0075] The weight content of each component in the mulberry extract is:

[0076] The weight content of each component in the mulberry extract is:

[0077] The weight content of each component in the mulberry extract is:

[0078] The weight content of each component in the mulberry extract is:

[0079] The weight content of each component in the mulberry extract is:

[0080] The weight content of each component in the mulberry extract is:

[0081] The weight content of each component in the mulberry extract is:

[0082] The weight content of each component in the mulberry extract is:

[0083] The weight content of each component in the mulberry extract is:

[0084] The weight content of each component in the mulberry extract is:

[0085] The weight content of each component can be any combination of the above values.

[0086] Optionally, the alkaloids include 1-deoxynojirimycin (DNJ), N-methyl-1-deoxynojirimycin (N-methly-1-deoxynojirimycin), fagomine (FAG), 3-epi-fagomine, 1,4-dideoxy-1,4-imino-D-arabinitol (DAB), calystegin B2, calystegin C1, 2-oxy-(α-D- One or more of 2-O-(α-D-galactopyranosyl)-1-deoxynojirimycin, 6-O-(β-D-glucopyranosyl)-1-deoxynojirimycin, and 1,4-dideoxy-1,4-imino-(2-O-β-D-glucopyranosyl)-D-arabinitol.

[0087] Wherein, optionally, the weight percentage of DNJ is not less than 50% (optionally 60-99%) of the total alkaloids.

[0088] Optionally, the heavy metal content of the mulberry extract does not exceed 10 ppm.

[0089] The drug is in the form of an oral dosage form; optionally, the drug is in the form of a tablet, a capsule, an oral solution, an oral emulsion, a pill, a granule, a syrup, and a powder.

[0090] In the present invention, the mulberry extract can be provided in the form of commercially available Morus alkaline tablets (National Medicine Standard No. Z20200002).

[0091] Alternatively, the mulberry extract can be prepared according to the method described in CN 110393738A. In one embodiment, the preparation of the mulberry extract comprises the following steps: preparing a crude extract of a moraceae plant; optionally, separating the extract with a cationic resin and / or an anionic resin; optionally, subjecting the resin effluent to alcohol precipitation and collecting the supernatant; and optionally, concentrating and / or drying the supernatant.

[0092] Optionally, in the present invention, the preparation of the mulberry extract comprises the following steps: 1) preparing a crude extract of a moraceae plant; 2) separating the crude extract through a cationic resin and / or an optional anionic resin to obtain a resin effluent; and optionally, step 3) subjecting the resin effluent of step 2) to alcohol precipitation and collecting the supernatant; and 4) concentrating and / or drying the supernatant. Optionally, the resin effluent of step 2) may be concentrated and / or dried before being subjected to alcohol precipitation.

[0093] Optionally, the plant of the Moraceae family is Morus multicaulis Perrott., Morus alba L., Morus atropurpurea Roxb., Morus mizuho Hotta, Morus wittiorum Hand Mazz., Morus laevigata Wall, Morus nigra Linn., Morus cathayana Hemsi., Morus serrata Roxb., Morus mongolica Schneid., Morus bombycis Koidz., Morus notabilis Schneid., Morus nigriformis Koidz., Morus yunnanensis Koidz., Morus australis Poir., Morus mongolica (Bur.) Schneid var. diabolica Koidz.), Morus alba Var. Pendula Dippel, Morus alba Var. Pendula Dippel, Morus alba, and mulberry varieties bred from the above mulberry species, or a combination of one or more of the following:

[0094] Optionally, the moraceae plant is one or more selected from the group consisting of Guangdong mulberry, Shandong mulberry, white mulberry, fine-toothed mulberry, mountain mulberry, or hybrid mulberry, and the hybrid mulberry can be Yuesang No. 11, Guisang You No. 62, or Sang Te You No. 2. Any part of the moraceae plant, including leaves, roots, branches, bark, buds, stems, and fruits, can be used, and mulberry branches, mulberry leaves, or white mulberry bark can be used.

[0095] In the present invention, the mulberry extract can be selected from mulberry branch extract, white mulberry bark extract, mulberry leaf extract or a mixed extract thereof.

[0096] In one embodiment, the mulberry extract is prepared according to the following steps: crushing the moraceae plant, heating and refluxing the extract with water and / or alcohol solution or acid water, the amount of solvent is 3-20 times that of the original medicinal material, repeating the extraction 1-3 times, combining the extracts, concentrating, applying a cation exchange resin, eluting with 0.2-3N ammonia water, applying the eluate to an anion exchange resin, collecting the non-adsorbed portion, adding ethanol, precipitating to remove impurities, concentrating and / or drying to obtain the extract.

[0097] In one embodiment, the mulberry extract is prepared according to the following steps: crushing the moraceae plant, heating and refluxing the extract with water and / or alcohol solution or acid water, the amount of solvent is 3-20 times that of the original medicinal material, repeating the extraction 1-3 times, combining the extracts, concentrating, applying a cation exchange resin, eluting with 0.2-3N ammonia water, applying the eluate to an anion exchange resin, collecting the non-adsorbed portion, concentrating and / or drying to obtain the extract.

[0098] In one embodiment, the mulberry extract is prepared according to the following steps: crushing the moraceae plant, heating and refluxing the extract with water and / or alcohol solution or acid water, the amount of solvent is 3-20 times that of the original medicinal material, repeating the extraction 1-3 times, combining the extracts, concentrating, applying a cation exchange resin, eluting with 0.2-3N ammonia water, concentrating and / or drying the eluate to obtain the extract.

[0099] In one embodiment, the mulberry extract is prepared according to the following steps: crushing the moraceae plant, heating and refluxing the extract with water, the amount of solvent is 3-20 times (optionally 4-15 times, further optionally 4-12 times) of the original medicinal material, repeating the extraction 1-3 times (the extraction time can be 0.5-3 hours each time, further optionally 1-2 hours each time), combining the extracts, concentrating, applying a cation exchange resin, eluting with 0.2-3N ammonia water, applying the eluate to an anion exchange resin, collecting the non-adsorbed portion (i.e., the anion resin effluent), adding ethanol, precipitating to remove impurities, concentrating and / or drying to obtain an extract.

[0100] Optionally, the crude extract after the concentration treatment can also be subjected to alcohol precipitation treatment before the resin separation treatment in step 2). During the alcohol precipitation treatment, ethanol is added to the crude extract, stirred and mixed, and the stirring is stopped and allowed to stand for a certain time to precipitate the insoluble matter therein. Optionally, the volume mass ratio of the added ethanol to the plant raw material is 0.2-20 times, optionally 0.4-10 times, in terms of L / kg. Further optionally, an alcohol precipitation tank is used for alcohol precipitation treatment. Optionally, the stirring speed in the alcohol precipitation treatment is 10-600rpm, optionally 40-500rpm, further optionally 80-400rpm or 300rpm.

[0101] Optionally, after the cationic resin is loaded into the column, it is activated in the order of washing with an acidic solution, washing with an alkaline solution, and washing with an acidic solution. Optionally, the alkaline solution is washed until the pH of the eluate is 8.0-9.5, optionally 8.5-9.5; optionally, the alkaline solution is selected from aqueous ammonia solution, sodium hydroxide solution, potassium hydroxide solution, or sodium carbonate solution; optionally, the concentration of the alkaline solution is 0.5-4 mol / L, optionally 1-2 mol / L. Optionally, the acidic solution is washed until the pH of the eluate is 3.0-7.0, optionally 4.5-6.5. Optionally, the acidic solution is selected from hydrochloric acid solution, phosphoric acid solution, or sodium hydrogen phosphate-citrate buffer; optionally, the concentration of the acidic solution is 0.5-4 mol / L, optionally 1.5-2 mol / L. Optionally, after the final acidic solution wash, the cationic resin can be rinsed with 3-5 column volumes of deionized water.

[0102] Optionally, the cationic resin is a combination of one or more of 732 strong acid styrene cation exchange resin, 002SC strong acid styrene cation exchange resin, 734 strong acid styrene cation exchange resin, D001 macroporous strong acid styrene cation exchange resin or D113 macroporous weak acid cation exchange resin and D254 macroporous strong basic quaternary ammonium cation exchange resin.

[0103] Optionally, the weight ratio of the cationic resin to the plant raw material is 1:1-30 (optionally, 1:1-25, 1:2-20, 1:2-15, 1:2-10, 1:2-7, 1:2-3).

[0104] After the crude plant extract is loaded onto the cationic resin, the loaded cationic resin is eluted with an eluent. Optionally, the eluent is a salt solution or alkaline solution containing cations, and can be selected from one or more of sodium chloride, ammonium chloride, ammonium sulfate, ammonium nitrate, ammonia water, potassium chloride, and sodium hydroxide.

[0105] Optionally, the concentration of the eluent is 0.04-5 mol / L (optionally 0.5-2.5 mol / L, 0.2-3 mol / L, further optionally 0.5-2.5 mol / L).

[0106] Optionally, the eluent flow rate is 1-15 BV / h (optionally 5-10 BV / h, further optionally 5-6 BV / h).

[0107] Optionally, the weight of the eluent used for cationic resin separation is 0.1-30 times the weight of the plant raw material input. Optionally, the eluent is used in an amount of 0.5-10 times the weight of the plant raw material input for elution.

[0108] The collection starting point can be determined according to the pH of the cationic resin effluent. For example, when an alkaline solution such as aqueous ammonia is used for elution, the eluate is collected when the pH of the cationic column effluent is detected to be greater than 7, or the collection starting point of the effluent is determined based on a color development or precipitation reaction. Optionally, when the volume of the collected liquid reaches 0.1-10 times (further optionally, 0.2-5 times) the weight of the plant raw material fed, collection is stopped, and the collected liquid is optionally purified by an anion column.

[0109] When purifying by anion column, optionally, after the anion resin is loaded into the column, activation is performed in the order of washing with alkaline solution, washing with acidic solution, and washing with alkaline solution.

[0110] Optionally, washing with an alkaline solution until the pH of the eluate is 8.0-9.5, optionally 8.5-9.5;

[0111] Optionally, the alkaline solution is selected from ammonia solution, sodium hydroxide solution, potassium hydroxide solution or sodium carbonate solution; optionally, the concentration of the alkaline solution is 0.5-4 mol / L, optionally 1-2 mol / L.

[0112] Optionally, the acidic solution is washed until the pH of the eluate is 3.0-7.0, optionally 4.5-6.5. Optionally, the acidic solution is selected from hydrochloric acid solution, phosphoric acid solution, disodium hydrogen phosphate-citric acid buffer, and optionally, the concentration of the acidic solution is 0.5-4 mol / L, optionally 1-2 mol / L.

[0113] Optionally, the anion resin is a combination of one or more of 711 type strong basic styrene anion resin, 717 type strong basic styrene anion exchange resin, D201 type macroporous strong basic styrene anion exchange resin or D218 type macroporous strong basic acrylic anion exchange resin, D301-G type macroporous weak acid styrene anion exchange resin and D301 type macroporous weak basic styrene anion exchange resin.

[0114] Optionally, the weight ratio of the anion resin to the plant raw material is 1:1-80 (optionally, 1:1-64, 1:1-32, 1:1-24, 1:5-16, 1:3).

[0115] When the liquid flows out of the anion resin, collection begins. Optionally, collection is stopped when the volume of the collected liquid reaches 0.05-10 times (optionally, 0.1-5 times) the weight of the plant raw material input.

[0116] Optionally, the weight ratio of ethanol used in the alcohol precipitation treatment to the plant raw material is 1:4-600 (optionally 1:20-300, further optionally 1:20-50, 1:40, 1:80, 1:22). During the alcohol precipitation treatment, the stirring speed is 10-600 rpm (optionally 40-500 rpm, 80-400 rpm). The alcohol precipitation treatment time is 12-24 hours.

[0117] Furthermore, before the alcohol precipitation treatment, the anion resin effluent is subjected to a step of centrifugal impurity removal or microfiltration membrane filtration, followed by concentration by a reverse ion osmosis membrane. The specific gravity of the concentrated liquid can be 1.0-1.3, optionally 1.1-1.25.

[0118] Optionally, the drug further comprises a pharmaceutically acceptable carrier. The carrier is an inactive ingredient that is non-toxic to the human body and is consistent with the route of administration or mode of administration. The carrier can be a solid or liquid excipient. Solid excipients include, for example, microcrystalline cellulose, mannitol, lactose, pregelatinized starch, low-substituted hydroxypropyl cellulose, cross-linked polyvinylpyrrolidone, sodium carboxymethyl starch, aspartame, calcium hydrogen phosphate, sodium lactate, poloxamer, sodium lauryl sulfate, sodium carboxymethyl cellulose, gelatin, xanthan gum, povidone, starch, magnesium stearate, sodium carboxymethyl starch, and talc; liquid excipients include, for example, water, ethanol, syrup, and glycerol.

[0119] In the present invention, the mulberry extract is referred to as SZ-A. Beneficial effects:

[0120] Mulberry extract can improve infertility, including male sexual dysfunction and male reproductive disorders. Mulberry extract can increase total sperm count, motile sperm count, and sperm motility, improve penile tissue damage, and significantly increase testosterone levels in testicular tissue. It also significantly increases the expression of the rate-limiting enzyme STAR, the key enzyme in testosterone synthesis CYP17A1, and the androgen receptor AR. It also significantly improves the structure of the seminiferous tubules and the state of interstitial cells in the testicles, reduces inflammatory cell infiltration, and increases the number and number of interlayers between spermatogenic cells at all levels, resulting in tighter connections and more orderly arrangement. BRIEF DESCRIPTION OF THE DRAWINGS

[0121] FIG1 is the results of (A) total sperm count and (B) motile sperm count of male SD rats in each test group of Experimental Example 1;

[0122] Figure 2 shows the sperm count, motile sperm count, and sperm motility of GAN-induced male mice in each experimental group of Experimental Example 2 (n=5, data are "mean ± standard error", and the differences between the groups were compared using one-way analysis of variance. Compared with the Model group, *P<0.05, **P<0.01);

[0123] Figure 3 shows the testosterone levels in testicular tissue of GAN-induced male mice in each experimental group of Experimental Example 2 (n=10, data are "mean ± standard error", and the differences between the groups were compared using one-way analysis of variance. Compared with the Model group, *P<0.05, **P<0.01);

[0124] Figure 4 shows the STAR levels in the testicular tissue of GAN-induced male mice in each experimental group of Experimental Example 2 (n=3, data are "mean ± standard error", and the differences between the groups were compared using one-way analysis of variance. Compared with the Model group, *P<0.05, **P<0.01);

[0125] Figure 5 shows the testicular CYP17A1 levels of GAN-induced male mice in each experimental group of Experimental Example 2 (n=3, data are "mean ± standard error", and the differences between the groups were compared using one-way analysis of variance. Compared with the Model group, *P<0.05, **P<0.01);

[0126] Figure 6 shows the AR levels in testicular tissue of GAN-induced male mice in each experimental group of Experimental Example 2 (n=3, data are "mean ± standard error", and the differences between the groups were compared using one-way analysis of variance. Compared with the Model group, *P<0.05, **P<0.01);

[0127] FIG7 is a HE staining morphology of testicular tissue of GAN-induced male mice in each experimental group of Experimental Example 2;

[0128] Figure 8 shows the number of spermatogenic cells at all levels in the testicular tissue of GAN-induced male mice in each experimental group of Experimental Example 2 (data are "mean ± standard error", and the differences between the groups were compared using one-way analysis of variance. Compared with the Model group, *P<0.05, **P<0.01);

[0129] FIG9 shows the body weight results of HFD-induced obese C57BL / 6J mice in each experimental group of Experimental Example 3;

[0130] Figure 10 shows the sperm count, motile sperm count, and sperm motility of HFD-induced obese C57BL / 6J mice in each experimental group of Experimental Example 3; compared with the Model group, *P<0.05, **P<0.01;

[0131] FIG11 shows the testicular index results of HFD-induced obese C57BL / 6J mice in each experimental group of Experimental Example 3; compared with the Model group, *P<0.05, **P<0.01;

[0132] Figure 12 is a HE staining image of the corpus cavernosum structure of the penile tissue in the HFD diet-induced C57BL / 6J obese reproductive disorder mouse model in Experimental Example 3; compared with the Model group, *P<0.05, **P<0.01, ***P<0.001;

[0133] Figure 13 shows the sperm count, motile sperm count, and sperm motility of mice in each experimental group of Experimental Example 4; compared with the Model group, *P<0.05, **P<0.01;

[0134] 14 is a HE staining of the seminiferous tubule structure of the testicular tissue of mice in each experimental group of Experimental Example 4;

[0135] Figure 15 is a graph showing the number of spermatogenic cells at each level and the total number of spermatogenic cells in mice in each experimental group of Experimental Example 4; compared with the Model group, *P<0.05, **P<0.01;

[0136] Figure 16 is a HE staining image of the corpus cavernosum structure of the penile tissue of mice in each experimental group of Experimental Example 4; compared with the Model group, *P<0.05, **P<0.01, ***P<0.001;

[0137] FIG17 is the result of Experimental Example 5: the effect of SZ-A on PA (palmitic acid)-induced Leydig cell damage;

[0138] Figure 18 shows the results of Experiment 6: Effect of SZ-A on high glucose-induced cavernous endothelial cell injury; compared with the high glucose group, *P<0.05, **P<0.01, ***P<0.001;

[0139] Figure 19 is the result of Experimental Example 7: Effect of SZ-A on high glucose-induced cavernous endothelial cell injury 2; compared with the high glucose group, *P<0.05, **P<0.01, ***P<0.001;

[0140] FIG20 is the result of observing endothelial cell mitochondria under a fluorescence microscope in Experimental Example 8;

[0141] FIG21 is a graph showing the IIEF-15 scoring results of the clinical experiment of Experimental Example 9. DETAILED DESCRIPTION

[0142] The present invention is further illustrated below through specific experimental examples. It should be understood that the following experiments are only used to illustrate the technical solutions of the present invention and are not intended to limit the scope of protection of the present invention.

[0143] The mulberry extract of the present invention can be prepared by the following method:

[0144] 1. Preparation Example of Mulberry Extract

[0145] Preparation Example 1 of Mulberry Extract

[0146] 1000 kg of fresh mulberry branches (fine-toothed mulberry, Yuesang No. 11) were crushed and added to 4000 L of water. The extracts were then heated and refluxed for 2 hours. The combined extracts were filtered to remove insoluble matter to obtain a crude extract. The crude extract was heat-concentrated to a solids content of 4% by weight and then heated to 50°C to serve as the loading solution for a cationic resin column.

[0147] A 150 kg column of D113 macroporous weakly acidic phenyl propylene-based cationic resin was loaded and washed with 2 mol / L hydrochloric acid until the eluate pH reached 4.5; then with 1 mol / L sodium hydroxide until the eluate pH reached 8.5; and finally with 2 mol / L hydrochloric acid until the eluate pH reached 4.5. The column was then rinsed with 5 column volumes of deionized water to complete activation. The concentrated extract was loaded and then eluted with 1000 L of 2.5 mol / L ammonia at a rate of 6 BV / h. The eluate from the cation column was collected when the pH was >7. When the collected solution reached 900 L, the collection was stopped and the collected solution was directly passed through an anion column for purification.

[0148] Activate the column using 62.5 kg of D218 macroporous, strongly basic acrylic anion resin. Elute with 1.5 mol / L sodium hydroxide solution until the eluate has a pH of 9.0. Elute with 1.5 mol / L hydrochloric acid solution until the eluate has a pH of 3.5. Finally, elute with 1.5 mol / L sodium hydroxide solution until the eluate has a pH of 9.0. The collected eluate from the cationic resin is loaded onto the anionic resin, and the effluent is collected until 920 L of effluent is reached.

[0149] The collected liquid was centrifuged to remove impurities and then concentrated using a reverse ion osmosis membrane. The concentrated liquid had a specific gravity of 1.25 and was transferred to an alcohol precipitation tank. 25 L of anhydrous ethanol was added with a stirring paddle at 500 rpm. After the ethanol addition was complete, stirring was stopped and the mixture was allowed to settle for 24 hours. The supernatant was collected and concentrated under reduced pressure to obtain a mulberry branch extract (i.e., mulberry extract SZ-A).

[0150] The mulberry branch extract contains 52% by weight of alkaloids, 22% by weight of polysaccharides, 0.8% by weight of flavonoids, and 20% by weight of amino acids. Among the alkaloids, the content of 1-DNJ is 60%, FAG is 17%, and DAB is 15%.

[0151] Preparation Example 2 of Mulberry Extract

[0152] Take 10kg of fresh mulberry branches (Sang Teyou No. 2), crush them, add 150L of water, add them twice, extract them by decoction for 3h each time, combine the extracts, and filter to remove insoluble matter. The extract is hot concentrated until the solid content reaches 8%, then transferred to an alcohol precipitation tank, and 2367.9g of absolute ethanol (3L) is added under a stirring paddle at 300rpm. After the ethanol addition is complete, stop stirring, precipitate with alcohol for 24h, and take the supernatant as the loading liquid for a cationic resin column. Use 5kg of 002SC type strong acid styrene cationic resin to fill the column, and activate the cationic resin according to the method of Preparation Example 1. Load the extract after concentrated alcohol precipitation, then use 100L of 5mol / L potassium chloride to elute, with an elution rate of 5BV / h, and detect the effluent with 20% silicotungstic acid. Start collecting when a white precipitate is generated. Stop collecting when the collected liquid reaches 25L, and the collected liquid is directly purified by anion column.

[0153] A 10 kg column of 711 strong base styrene anion resin was loaded and activated according to the method of Preparation Example 1. The collected cationic resin eluate was loaded onto the anion resin, and the effluent was collected until the effluent reached 15 L. The collected eluate was reloaded onto the cationic resin and separated twice more using the cationic resin and anion resin, respectively, according to the above method.

[0154] The collected liquid obtained after three column separations was centrifuged for impurity removal and then concentrated using a reverse ion osmosis membrane. The concentrated liquid had a specific gravity of 1.25 and was transferred to an alcohol precipitation tank. 125 g of anhydrous ethanol was added under a stirring paddle at 1000 rpm. After the ethanol addition was complete, stirring was stopped and the mixture was allowed to precipitate with alcohol for 24 hours. The supernatant was then collected and concentrated under reduced pressure to obtain an extract. Separately, fresh Morus alba bark and mulberry leaf (Sang Teyou No. 2) were extracted using the same extraction method and parameters as described above.

[0155] The obtained mulberry branch extract has an alkaloid content of 98%, a polysaccharide content of 0.2%, a flavonoid content of 0.05%, and an amino acid content of 0. Among the alkaloids, the content of 1-DNJ is 99%, FAG is 0.5%, and DAB is 0.4%.

[0156] The obtained Morus alba bark extract contains 95% alkaloids, 2% polysaccharides, 0.1% flavonoids, and 1% amino acids. Among the alkaloids, the content of 1-DNJ is 96%, FAG is 1.5%, and DAB is 1.4%.

[0157] The mulberry leaf extract contains 90% alkaloids, 4% polysaccharides, 0.1% flavonoids, and 3% amino acids. Among the alkaloids, the content of 1-DNJ is 91%, FAG is 3.1%, and DAB is 2.8%.

[0158] Preparation Example 3 of Mulberry Extract

[0159] 1000 kg of fresh mulberry branches (Mulberry twigs) were crushed, added to 11,500 L of water, and heated under reflux for 2 hours. The extracts were combined and filtered to remove insoluble matter to obtain a crude extract. This crude extract was first centrifuged to remove impurities and then concentrated using a counter-ion permeation membrane to a solids content of 1% by weight. This was then used as the loading solution for the cationic resin column.

[0160] A column was loaded with 300 kg of D001 macroporous, strongly acidic styrene-based cationic resin, which was activated according to the method of Preparation Example 1. The concentrated crude extract was loaded and eluted with 5000 L of 0.04 mol / L ammonium nitrate at a rate of 5 BV / h. The effluent was tested with 20% silicotungstic acid. Collection began when a white precipitate formed and stopped when the collected solution reached 1000 L.

[0161] The collected liquid obtained after the cationic column separation is concentrated by nanofiltration membrane and concentrated under reduced pressure to obtain an extract concentrate.

[0162] The obtained mulberry branch extract has an alkaloid content of 15%, a polysaccharide content of 20%, a flavonoid content of 7%, and an amino acid content of 45%. Among the alkaloids, the content of 1-DNJ is 55%, FAG is 23%, and DAB is 10%.

[0163] Preparation Example 4 of Mulberry Extract

[0164] Take 333 kg of dry mulberry branches (Yue Sang No. 11), crush them, add 4000 L of water, and extract them twice by heating reflux method, each time reflux for 1 hour, combine the extracts, filter, and concentrate the extracts to 1 kg of crude drug / L.

[0165] A 150 kg column of D113 macroporous weakly acidic phenyl propylene-based cationic resin was loaded and washed with 2 mol / L hydrochloric acid until the eluate pH reached 4.5; then with 1 mol / L sodium hydroxide until the eluate pH reached 8.5; and finally with 2 mol / L hydrochloric acid until the eluate pH reached 4.5. The column was then rinsed with 5 column volumes of deionized water to complete activation. The concentrated extract was loaded and then eluted with 1000 L of 2.5 mol / L ammonia at a rate of 6 BV / h. The eluate from the cation column was collected when the pH was >7. When the collected solution reached 900 L, the collection was stopped and the collected solution was directly passed through an anion column for purification.

[0166] Activate the column using 125 kg of D218 macroporous, strongly basic acrylic anion resin. Elute with 1.5 mol / L sodium hydroxide solution until the eluate has a pH of 9.0. Elute with 1.5 mol / L hydrochloric acid solution until the eluate has a pH of 3.5. Finally, elute with 1.5 mol / L sodium hydroxide solution until the eluate has a pH of 9.0. The collected cationic resin eluate is loaded onto the anion resin, and the effluent with a pH greater than 8 is collected until the effluent reaches 870 L.

[0167] The collected liquid after anion column separation was filtered through a microfiltration membrane to remove impurities and then concentrated using a counter-ion osmosis membrane. The concentrated liquid had a specific gravity of 1.1 and was transferred to an alcohol precipitation tank. 15 kg of anhydrous ethanol was added with a stirring paddle at 400 rpm. Stirring was stopped after the ethanol addition, and the mixture was allowed to settle for 24 hours. The supernatant was collected and concentrated under reduced pressure to obtain a mulberry branch extract. The sample contained 80% alkaloids by weight, 5% polysaccharides by weight, 0.1% flavonoids by weight, and 4% amino acids by weight. Among the alkaloids, 1-DNJ accounted for 75%, FAG for 12%, and DAB for 10%.

[0168] Preparation Example 5 of Mulberry Extract

[0169] Take 400 kg of dry mulberry branches (Yue Sang No. 11), crush them, add 4000 L of water, and extract them twice by heating reflux method, each time reflux for 1 hour, combine the extracts, filter, and concentrate the extracts to 1 kg of crude drug / L.

[0170] Activate the column using 62.5 kg of D218 macroporous, strongly basic acrylic anion resin. Elute with 1.5 mol / L sodium hydroxide solution until the eluate has a pH of 9.0; then with 1.5 mol / L hydrochloric acid solution until the eluate has a pH of 3.5; and finally with 1.5 mol / L sodium hydroxide solution until the eluate has a pH of 9.0. The collected extract concentrate was loaded onto the anion resin, and the effluent was collected.

[0171] The collected solution after anion column separation was filtered through a microfiltration membrane to remove impurities, then concentrated using a counter-ion permeation membrane. Further vacuum concentration and drying yielded a mulberry twig extract. The sample contained 3% by weight of alkaloids, 70% by weight of polysaccharides, 10% by weight of flavonoids, and 10% by weight of amino acids. Among the alkaloids, the content of 1-DNJ was 68%, FAG was 17%, and DAB was 8%.

[0172] Preparation Example 6 of Mulberry Extract

[0173] 1500 kg of fresh mulberry branches (fine-toothed mulberry, Yuesang No. 11) were crushed and added to 6000 L of water. The extracts were heated and refluxed for 2 hours. The combined extracts were filtered to remove insoluble matter to obtain a crude extract. The crude extract was heat-concentrated to a solids content of 4% by weight and then heated to 50°C as the loading solution for a cationic resin column.

[0174] A 100 kg column of D113 macroporous weakly acidic phenyl propylene-based cationic resin was loaded and washed with 2 mol / L hydrochloric acid until the eluate pH reached 4.5; then with 1 mol / L sodium hydroxide until the eluate pH reached 8.5; and finally with 2 mol / L hydrochloric acid until the eluate pH reached 4.5. The column was then rinsed with 5 column volumes of deionized water to complete activation. The concentrated extract was loaded and then eluted with 1000 L of 2.5 mol / L ammonia at a rate of 6 BV / h. The eluate from the cation column was collected when the pH was >7. When the collected solution reached 900 L, the collection was stopped and the collected solution was directly passed through an anion column for purification.

[0175] A column was loaded with 62.5 kg of D218 macroporous, strongly alkaline acrylic anion resin. The column was washed with a 1.5 mol / L sodium hydroxide solution until the eluate had a pH of 9.0; then washed with a 1.5 mol / L hydrochloric acid solution until the eluate had a pH of 3.5; and finally washed with a 1.5 mol / L sodium hydroxide solution until the eluate had a pH of 9.0. Activation was completed. The collected cationic resin eluate was loaded onto an anionic resin, and the effluent was collected until the effluent reached 870 L. The effluent was concentrated under reduced pressure to obtain a mulberry branch extract concentrate, which contained 30% by weight alkaloids, 35% by weight polysaccharides, 2% by weight flavonoids, and 25% by weight amino acids. Among the alkaloids, the content of 1-DNJ was 62%, FAG was 20%, and DAB was 13%.

[0176] Preparation Example 7 of Mulberry Extract

[0177] 1000 kg of fresh mulberry branches (fine-toothed mulberry, Yuesang No. 11) were crushed and added to 4000 L of water. The extracts were then heated and refluxed for 2 hours. The combined extracts were filtered to remove insoluble matter to obtain a crude extract. The crude extract was heat-concentrated to a solids content of 4% by weight and then heated to 50°C to serve as the loading solution for a cationic resin column.

[0178] A 100 kg column of D113 macroporous weakly acidic phenyl propylene-based cationic resin was loaded and washed with 2 mol / L hydrochloric acid until the eluate pH reached 4.5; then with 1 mol / L sodium hydroxide until the eluate pH reached 8.5; and finally with 2 mol / L hydrochloric acid until the eluate pH reached 4.5. The column was then rinsed with 5 column volumes of deionized water to complete activation. The concentrated extract was loaded and then eluted with 1000 L of 2.5 mol / L ammonia at a rate of 6 BV / h. The eluate from the cation column was collected when the pH was >7. When the collected solution reached 900 L, the collection was stopped and the collected solution was directly passed through an anion column for purification.

[0179] A 62.5 kg column of D218 macroporous, strongly alkaline acrylic anion resin was loaded and washed with a 1.5 mol / L sodium hydroxide solution until the eluate had a pH of 9.0; then with a 1.5 mol / L hydrochloric acid solution until the eluate had a pH of 3.5; and finally with a 1.5 mol / L sodium hydroxide solution until the eluate had a pH of 9.0. Activation was completed. The collected cationic resin eluate was loaded onto an anionic resin, and the effluent was collected until the effluent reached 870 L. The effluent was concentrated under reduced pressure to obtain a mulberry branch extract concentrate, which contained 40% by weight alkaloids, 25% by weight polysaccharides, 0.5% by weight flavonoids, and 25% by weight amino acids. Among the alkaloids, the content of 1-DNJ was 57%, FAG was 24%, and DAB was 16%.

[0180] Preparation Example 8 of Mulberry Extract

[0181] Take 333 kg of dry mulberry branches (Yue Sang No. 11), crush them, add 4000 L of water, and extract them twice by heating reflux method, each time reflux for 1 hour, combine the extracts, filter, and concentrate the extracts to 1 kg of crude drug / L.

[0182] A 150 kg column of D113 macroporous weakly acidic phenyl propylene-based cationic resin was loaded and washed with 2 mol / L hydrochloric acid until the eluate pH reached 4.5; then with 1 mol / L sodium hydroxide until the eluate pH reached 8.5; and finally with 2 mol / L hydrochloric acid until the eluate pH reached 4.5. The column was then rinsed with 5 column volumes of deionized water to complete activation. The concentrated extract was loaded and then eluted with 1000 L of 2.5 mol / L ammonia at a rate of 6 BV / h. The eluate from the cation column was collected when the pH was >7. When the collected solution reached 900 L, the collection was stopped and the collected solution was directly passed through an anion column for purification.

[0183] Activate the column using 62.5 kg of D218 macroporous, strongly basic acrylic anion resin. Elute with 1.5 mol / L sodium hydroxide solution until the eluate has a pH of 9.0. Elute with 1.5 mol / L hydrochloric acid solution until the eluate has a pH of 3.5. Finally, elute with 1.5 mol / L sodium hydroxide solution until the eluate has a pH of 9.0. The collected cationic resin eluate is loaded onto the anion resin, and the effluent with a pH greater than 8 is collected until the effluent reaches 870 L.

[0184] The collected liquid after anion column separation was filtered through a microfiltration membrane to remove impurities and then concentrated using a counter-ion permeation membrane. The concentrated liquid had a specific gravity of 1.1 and was transferred to an alcohol precipitation tank. 15 kg of anhydrous ethanol was added with a stirring paddle at 400 rpm. Stirring was stopped after the ethanol addition, and the mixture was allowed to settle for 24 hours. The supernatant was collected and concentrated under reduced pressure to obtain a mulberry twig extract. The sample contained 63% alkaloids by weight, 23% polysaccharides by weight, 1% flavonoids by weight, and 5% amino acids by weight. Among the alkaloids, the content of 1-DNJ was 61.9%, FAG was 16.6%, and DAB was 11.1%.

[0185] Preparation Example 9 of Mulberry Extract

[0186] 1000 kg of fresh mulberry branches (Yue Sang No. 11) were crushed and added to 4000 L of water. The extracts were then heated and refluxed for 2 hours. The combined extracts were filtered to remove insoluble matter to obtain a crude extract. The crude extract was then heat-concentrated to a solids content of 4% and maintained at 50°C as the loading solution for a cationic resin column.

[0187] A 120 kg column of D113 macroporous weakly acidic phenyl propylene-based cationic resin was loaded and washed with 2 mol / L hydrochloric acid until the eluate pH reached 4.5; then with 1 mol / L sodium hydroxide solution until the eluate pH reached 8.5; then with 2 mol / L hydrochloric acid until the eluate pH reached 4.5; and finally with 5 column volumes of deionized water to complete the activation. The concentrated extract was loaded and then eluted with 1000 L of 2.5 mol / L ammonia at a rate of 6 BV / h. The eluate from the cation column was collected when the pH was > 7. Collection was stopped when 900 L of the collected solution reached and the collected solution was directly passed through an anion column for purification.

[0188] Activate the column using a 45 kg column of D218 macroporous, strongly basic acrylic anion resin. Elute with 1.5 mol / L sodium hydroxide solution until the eluate has a pH of 9.0. Elute with 1.5 mol / L hydrochloric acid solution until the eluate has a pH of 3.5. Elute with 1.5 mol / L sodium hydroxide solution until the eluate has a pH of 9.0. The collected cation resin eluate is loaded onto the anion resin, and the effluent is collected until 870 L of effluent are reached.

[0189] After anion column separation, the collected liquid was filtered through a microfiltration membrane to remove impurities and then concentrated using a counter-ion permeation membrane. The concentrated liquid had a specific gravity of 1.1 and was transferred to an alcohol precipitation tank. 15 kg of anhydrous ethanol was added with a stirring paddle at 300 rpm. After the ethanol addition was complete, stirring was stopped. The alcohol precipitation was allowed to proceed for 24 hours. The supernatant was collected and concentrated under reduced pressure to obtain an extract. The sample contained 70% alkaloids by weight, 20% polysaccharides by weight, 0.6% flavonoids by weight, and 5% amino acids by weight. Among the alkaloids, the content of 1-DNJ was 70%, FAG was 13%, and DAB was 10%.

[0190] 2. Efficacy test of mulberry extract

[0191] Experimental Example 1: Effect of mulberry extract on spermatogenesis in normal rats

[0192] 1. Test samples

[0193] The mulberry extract of Preparation Example 1 was prepared.

[0194] 2. Preparation of dosing solution

[0195] First, weigh a certain amount of the test sample, place it in an appropriate beaker, add an appropriate amount of solvent, and ultrasonically (300W, 40KHZ) for about 10 minutes until it is completely dissolved. After standing at room temperature, transfer it into a measuring cylinder to the required volume to obtain the preparation solution of the required concentration.

[0196] 3. Animal Medication and Material Collection

[0197] Forty 5- to 7-week-old male Sprague-Dawley rats (138-183 g) were randomly divided into a vehicle control group and three mulberry extract treatment groups (dosages based on total alkaloids: 100 mg / kg / day, 200 mg / kg / day, and 500 mg / kg / day, respectively). The rats were gavage-administered once daily at a dose of 10 mL / kg. After 4 weeks of treatment, the rats were sacrificed, and the testes and epididymis were harvested.

[0198] 4. Detection Methods

[0199] Sperm count:

[0200] One side of the epididymis was placed in 1 mL of 0.9% sodium chloride injection and the epididymal tissue was minced. The cells were incubated at 37°C to prepare a sperm suspension. The sperm suspension was flushed into a blood cell counting chamber, and the number of sperm under the microscope was counted. The total sperm count was then calculated according to the dilution multiple.

[0201] Sperm motility test:

[0202] Preparation: Cut open the cauda epididymis with ophthalmic scissors. Mix the contents of the cauda epididymis with 0.9% sodium chloride injection to create a sperm suspension. Place two drops of the sperm suspension on a glass slide. This procedure should be performed at 37°C.

[0203] Observation: Record the motility of approximately 200 random sperm under a high-power microscope. Criteria for determining sperm motility: I. Good motility, with sperm swimming rapidly and in a straight line; II. Fair motility, with relatively active sperm; III. Poor motility, with sperm moving slowly and spinning in circles; IV. Dead sperm, with sperm morphology but no motility.

[0204] 5. Test results

[0205] Compared with the vehicle control group, the sperm count and motile sperm count of male rats in the mulberry extract groups at all doses increased (see Table 1 and Figure 1).

[0206] Table 1 Sperm function Note: Dilution method: Take one epididymis, mince it, add 1 mL of normal saline, remove 50 μL and dilute to 1 mL. Take another 100 μL and dilute to 1 mL again before counting under a microscope. Count the sperm (M) in the four large squares surrounding the hemocytometer. Calculate the formula: M / 0.4 × dilution factor × 1000 = sperm count / mL. Sperm motility = (grade I + grade II + grade III) / (grade I + grade II + grade III + grade IV) × 100%.

[0207] Table 1 and Figure 1 show that when mulberry extract was administered orally to healthy male SD rats for 4 consecutive weeks, the total sperm count and motile sperm count increased, indicating that mulberry extract can promote spermatogenesis in male rats.

[0208] Experimental Example 2: GAN-induced male mouse reproductive dysfunction model

[0209] 1. Test samples

[0210] Preparation Example 8 Mulberry Branch Extract (SZ-A)

[0211] 2. The experimental groups are as follows:

[0212] Table 2 Note: Gubra-Amylin NASH (GAN) is a high-fat, high-cholesterol, high-fructose feed containing 40% saturated fat, 22% fructose and 2% cholesterol.

[0213] 3. Experimental Methods

[0214] Fifty male C57BL / 6J mice in good health and with shiny fur were selected and fed a maintenance diet. They were then observed for one week of acclimatization. After ensuring that the animals were well adapted, modeling began. The model group was given a GAN (Research Diets D09100310) diet, while the control group continued to receive the maintenance diet for 20 weeks to establish a mouse model of male reproductive dysfunction. The mice were divided into five groups: a control group, a model group, and a low-, medium-, and high-dose group of mulberry twig extract (respectively designated SZ-AL, SZ-AM, and SZ-AH), with 10 mice in each group. Each experimental group was administered the drug according to the dosage and administration method in the table. The blank group and the model group were given an equal amount of normal saline. Samples were collected 21 days after administration for efficacy evaluation.

[0215] 4. Detection Methods

[0216] 1) Sperm count test:

[0217] After dissecting the mice, the epididymal tissue was taken from one side and placed in physiological saline (1 mL). The epididymis was minced and sperm was collected. The cells were then incubated in a shaking incubator at 37.6°C for 5 min, and the sperm count was immediately detected using a hemocytometer.

[0218] 2) Motile sperm count test:

[0219] Use ophthalmic scissors to cut open the cauda epididymis. Mix the contents of the cauda epididymis with 0.9% sodium chloride injection to prepare a sperm suspension. Place two drops of the sperm suspension on a glass slide. This procedure is performed at 37°C. Observe under a high-power microscope and record the swimming patterns of 200 random sperm. Calculate the number of motile sperm and sperm motility (sperm motility = number of motile sperm / number of sperm).

[0220] 3) Elisa was used to detect testosterone (T) levels.

[0221] 4) Western blot was used to detect the levels of androgen synthase (STAR, CYP17A1) and androgen receptor (AR).

[0222] 5) Pathology: HE staining was used to observe the structure of the seminiferous tubules, the morphology and number of spermatogenic cells in the testicular tissue, and the sperm maturation and spermatocyte grading.

[0223] 5. Results Analysis

[0224] (1) Total sperm count, motile sperm count, and sperm motility

[0225] The results in Figure 2 show that compared with the blank group, the total sperm count, motile sperm count and sperm motility in the model group were significantly reduced. After treatment with SZ-A, the low, medium and high doses of SZ-A groups could significantly increase the sperm count, motile sperm count and sperm motility, especially the improvement effect on the total sperm count, which basically returned to normal.

[0226] (2) Testosterone levels

[0227] The results in Figure 3 show that compared with the blank group, the testosterone level in the model group was significantly decreased. After treatment with SZ-A, the testosterone levels in the low, medium and high dose groups of SZ-A were significantly increased.

[0228] (3) STAR, CYP17A1, and AR levels

[0229] The results in Figures 4-6 show that the high-dose SZ-A group significantly increased the expression of the rate-limiting enzyme STAR in testosterone synthesis, the important enzyme CYP17A1 in testosterone synthesis, and the androgen receptor AR.

[0230] (4) Structure of seminiferous tubules in testicular tissue

[0231] The results in Figure 7 show that in the Blank group, the seminiferous tubules were tightly connected, spermatogenic cells at all levels were neatly arranged, and no obvious abnormalities were observed in the morphology of the cells. In the Model group, the seminiferous tubules were loosely connected, with interstitial cell vacuolation and inflammatory cell infiltration. Within the seminiferous tubules, the connections between spermatogenic cells at all levels were loose and disordered, with a reduced number of spermatogenic cell layers and some spermatogenic cells falling off. Compared with the Model group, all doses of SZ-A significantly improved the structure of the seminiferous tubules and the state of interstitial cells, reduced inflammatory cell infiltration, and increased the number and number of spermatogenic cells at all levels, with tight connections and neat arrangement.

[0232] (5) Number of spermatogenic cells

[0233] The results in Figure 8 show that compared with the blank group, the number of spermatogonia, primary spermatocytes, secondary spermatocytes, spermatids, and all spermatogenic cells in individual seminiferous tubules of mice in the Model group was significantly reduced; after treatment with SZ-A, the number of spermatogenic cells in each dose group was significantly increased in a dose-effect relationship. Among them, the improvement effect of the high-dose SZ-A group on the number of spermatogenic cells was close to that of the normal group.

[0234] Experimental Example 3: HFD diet-induced C57BL / 6J obese reproductive disorder mouse model. 45 healthy male C57BL / 6J mice aged 6 weeks were randomly divided into a normal group (Chow), a high-fat model group (HFD), and a mulberry extract (SZ-A), with 9 mice in each group. The mulberry extract was the mulberry extract prepared in Preparation Example 9. The mice in the normal group were fed with a basal feed, and the model group and the SZ-A group were fed with a high-fat diet (Research Diet, D12492, 60kcal% Fat); after feeding for 8 weeks, each experimental group was treated with the corresponding dose of the test drug for 6 consecutive weeks. During the treatment period, the high-fat diet was continued, and the body weight and food and water intake of the animals were monitored. The model group was given normal saline. After the experiment, samples were collected for efficacy evaluation.

[0235] Table 3 Experimental groups and drug administration methods

[0236] Detection indicators:

[0237] (1) The total sperm count, motile sperm count, and sperm motility of mice were determined by referring to Experimental Example 2.

[0238] (2) Mouse model penile tissue pathology: HE staining was used to observe the cavernous structure of the penile tissue and evaluate the improvement of the average thickness, connection density, separation degree and average intercept length of the sinusoidal space of the cavernous scaffold.

[0239] Results in Figures 9-12 show that in the model group, total sperm count, motile sperm count, and sperm motility decreased, while body weight increased significantly. Testicular index decreased, and the structure of the corpus cavernosum deteriorated, with numerous free-standing sponge scaffolds visible in the central region. The number of branches in the corpus cavernosum scaffolds was significantly reduced, and local sinusoidal spaces were fused and their area significantly increased. Compared with the model group, SZ-A significantly increased total sperm count, motile sperm count, and sperm motility, reduced body weight, and increased testicular index, which almost returned to normal levels. The sponge scaffolds within the corpus cavernosum showed a reticular distribution, with a slightly reduced number of branches, thicker and more compact scaffolds, and smaller interstitial spaces.

[0240] Experimental Example 4 STZ combined with high fat and high cholesterol induced Apoe diabetic sexual dysfunction model in mice

[0241] Thirty 8-week-old Apoe mice were adaptively fed for one week and then fed a high-fat, high-cholesterol diet (HFHC) for 5 weeks. On the 6th week, they were intraperitoneally injected with STZ (100 mg / kg / d) to establish a mouse model of lipid metabolism disorder. At the same time, 10 8-week-old C57 mice were adaptively fed for one week and then fed a normal diet for 5 weeks. On the 6th week, they were intraperitoneally injected with an equal amount of solvent once to serve as the normal control group.

[0242] After successful modeling, the model group was randomly divided into 3 groups, including the model group, the low (150 mg / kg / d) and high (300 mg / kg / d) dose groups of SZ-A of Preparation Example 8, and the drugs were administered according to the following table for 6 weeks, and then the samples were collected for efficacy evaluation.

[0243] Table 4 Animal grouping and drug administration

[0244] Detection indicators:

[0245] (1) The method for detecting the total sperm count, motile sperm count and sperm motility of mice was as described in Experimental Example 2.

[0246] pathology:

[0247] (2) Effects of SZ-A on testicular pathology: HE staining was used to observe the structure of the seminiferous tubules and the morphology and number of spermatogenic cells in the testicular tissue, and to observe sperm maturation and spermatocyte grading.

[0248] (3) HE staining was used to observe the results of the cavernous structure of the penile tissue and evaluate the improvement of the average thickness, connection density, separation degree and average intercept length of the spongy scaffold.

[0249] Result analysis:

[0250] (1) The results in Figure 13 show that the total sperm count, motile sperm count, and sperm motility of mice in the model group decreased. Compared with the model group, SZ-A significantly increased the total sperm count, motile sperm count, and sperm motility of mice.

[0251] (2) The results in Figure 14 show that the seminiferous tubules of the Blank group mice were tightly connected, the spermatogenic cells at all levels were neatly arranged, and no obvious abnormalities were observed in the morphology of the cells. The seminiferous tubules of the Model group mice were loosely connected, and the interstitial cells showed vacuolar degeneration and inflammatory cell infiltration. The spermatogenic cells at all levels in the seminiferous tubules were loosely connected and arranged in disorder, the number of spermatogenic cell layers decreased, and some spermatogenic cells fell off. Compared with the Model group, all doses of SZ-A significantly improved the structure of the seminiferous tubules and the state of the interstitial cells, reduced inflammatory cell infiltration, and increased the number and number of spermatogenic cells at all levels, with tight connections and neat arrangement.

[0252] The results in Figure 15 show that compared with the Blank group, the number of spermatogonia, primary spermatocytes, secondary spermatocytes, spermatids, and total spermatogenic cells in individual seminiferous tubules of mice in the Model group was significantly reduced. After treatment with SZ-A, the number of spermatogenic cells in each dose group was significantly increased in a dose-response relationship. Among them, the improvement effect of the high-dose SZ-A group on the number of spermatogenic cells was close to that of the normal group.

[0253] (3) HE staining to observe the structure of the corpus cavernosum of the penis. The results of pathological observation are shown in Figure 16. In the model group, the structure of the corpus cavernosum of the penis deteriorated, with a large number of free-like sponge scaffolds visible in the central part. The number of branches of the corpus cavernosum scaffolds was significantly reduced, and the local sinusoidal spaces were fused and the area was significantly increased. Compared with the model group, the sponge scaffolds in the corpus cavernosum of the penis in the SZ-A group were distributed in a reticular pattern. The number of branches of the corpus cavernosum scaffolds was slightly reduced, the scaffolds were thick and compact, and the scaffold gaps were smaller. The average thickness, connection density, separation degree and average intercept length of the sponge scaffolds were improved.

[0254] Experimental Example 5: Effect of SZ-A on PA (palmitic acid)-induced Leydig cell damage

[0255] 5*10 5 / mLTM3 cells (normal mouse testicular Leydig cells) were seeded in 6-well plates and cultured overnight. The cells were then divided into a blank group, a model group, and an SZ-A dosage group (specific groupings are as follows). 400 μmol / l PA was added to the model group and the SZ-A administration group, and the SZ-A group was given the drug. After incubation for 24 hours, the efficacy was evaluated.

[0256] Group 1: control group;

[0257] Group 2: Model group (PA group);

[0258] Group 3: drug administration group, the drug was 200 μg / mL of the mulberry branch extract in Preparation Example 1;

[0259] Group 4: drug administration group, the drug was 200 μg / mL of the mulberry branch extract prepared in Example 2;

[0260] Group 5: drug administration group, the drug was 200 μg / mL of the mulberry branch extract in Preparation Example 3;

[0261] Group 6: drug administration group, the drug was 200 μg / mL of the mulberry branch extract prepared in Example 4;

[0262] Group 7: drug administration group, the drug was 200 μg / mL of the mulberry branch extract in Preparation Example 5;

[0263] Group 8: drug administration group, the drug was 200 μg / mL of the mulberry branch extract in Preparation Example 7.

[0264] 1) Western blotting was used to detect the rate-limiting enzyme STAR in SZ-A testosterone synthesis, as well as the important enzymes CYP17A1 and 17β-HSD in testosterone synthesis.

[0265] The results in Figure 17 show that after PA induced Leydig cells, the expression of multiple androgen synthesis-related enzymes CYP17A1, STAR, and 17β-HSD in the model group was reduced. After intervention with different levels of SZ-A, the expression of androgen synthesis-related enzymes was increased.

[0266] Experimental Example 6 Effect of SZ-A on High Glucose-Induced Endothelial Cell Injury

[0267] 5000 / well RCCEC cells (cavernous endothelial cells) were seeded in 96-well plates and cultured overnight without serum. They were divided into control group, isotonic control group, model group and SZ-A dosage group (specific groups are as follows). The control group used endothelial cell-specific culture medium containing 5.55 mM glucose, the isotonic control group used endothelial cell-specific culture medium containing 5.55 mM glucose and supplemented with 24.45 mM mannitol, the model group and SZ-A treatment group used endothelial cell-specific culture medium containing 5.55 mM glucose and supplemented with 24.45 mM glucose (i.e., 30 mM glucose), and the SZ-A group was given drugs. After incubation for 72 h, cell viability was detected using CCK-8 reagent and the cell survival rate was calculated.

[0268] Group 1: Control group: 5.55 mM glucose

[0269] Group 2: isotonic control group: 5.55mM glucose + 24.45mM mannitol

[0270] Group 3: Model group: 30 mM glucose

[0271] Group 4: 30 mM glucose + 150 μg / ml SZ-A (based on the extract); SZ-A is the extract in Preparation Example 1;

[0272] Group 5: 30 mM glucose + 150 μg / ml SZ-A (based on the extract); SZ-A is the extract in Preparation Example 8;

[0273] Group 6: 30 mM glucose + 150 μg / ml SZ-A (based on the extract); SZ-A is the extract in Preparation Example 4;

[0274] Group 7: 30 mM glucose + 150 μg / ml SZ-A (based on the extract); SZ-A is the mulberry branch extract in Preparation Example 2;

[0275] The results in Figure 18 show that SZ-A can inhibit high glucose-induced cavernous endothelial cell damage and improve cell survival rate.

[0276] Experimental Example 7 Effect of SZ-A on High Glucose-Induced Endothelial Cell Injury

[0277] 3*10 5 / mL RCCEC cells (cavernous endothelial cells) were seeded in 6-well plates and cultured overnight without serum. They were then divided into a control group, an isotonic control group, a model group, and an SZ-A dosage group (specific groups are as follows). The control group received an endothelial-specific culture medium containing 5.55mM glucose, the isotonic control group received an endothelial-specific culture medium containing 5.55mM glucose and 24.45mM mannitol, the model group and the SZ-A administration group received an endothelial cell-specific culture medium containing 5.55mM glucose and supplemented with 24.45mM glucose (i.e., 30mM glucose), and the SZ-A group was given the drug. After incubation for 72h, the cells were collected and the intracellular nitric oxide content was detected.

[0278] Group 1: Control group: 5.55 mM glucose

[0279] Group 2: isotonic control group: 5.55mM glucose + 24.45mM mannitol

[0280] Group 3: Model group: 30 mM glucose

[0281] Group 4: 30 mM glucose + 150 μg / ml SZ-A (based on the extract); SZ-A is the extract in Preparation Example 3;

[0282] Group 5: 30 mM glucose + 150 μg / ml SZ-A (based on the extract); SZ-A is the extract in Preparation Example 1;

[0283] Group 6: 30 mM glucose + 150 μg / ml SZ-A (based on the extract); SZ-A is the extract in Preparation Example 4;

[0284] Group 7: 30 mM glucose + 150 μg / ml SZ-A (based on the extract); SZ-A is the mulberry branch extract in Preparation Example 2;

[0285] The results in Figure 19 show that SZ-A can inhibit high glucose-induced cavernous endothelial cell damage and improve the intracellular NO content.

[0286] Experimental Example 8 Effect of SZ-A on the Endothelial Cell Injury Model Induced by High Insulin

[0287] Cell culture: Human umbilical vein endothelial cells (HUVEC) were cultured in DMEM medium (10% FBS + 1% double antibody).

[0288] Cell passaging: 0.25% trypsinization, 10 5 Cells were seeded at a density of 1 / mL in a 12-well plate and cultured in a cell culture incubator at 37°C and 5% CO2. The culture medium was changed every 2-3 days, and the cells could be passaged again once they occupied approximately 80% of the field of view.

[0289] Model Construction: HUVEC cells were cultured overnight and randomly divided into six groups: a normal group, a model group, and an SZ-A group (Mulberry twig extract from Preparation Example 8, administered at doses of 100 μg / mL, 200 μg / mL, 300 μg / mL, and 400 μg / mL, based on total alkaloids). The normal group cells were treated with DMEM medium (10% FBS + 1% anti-hyperglycemic inhibitor), the model group (10% FBS + 1% anti-hyperglycemic inhibitor + 40 U insulin), and the SZ-A group (10% FBS + 1% anti-hyperglycemic inhibitor + 40 U insulin + the corresponding dose of SZ-A). After 24 hours of culture, mitochondrial membrane potential was measured by Mito-Tracker.

[0290] Mito-Tracker Deep Red FM (mitochondrial far-infrared fluorescent probe): When cells reach a certain density in a cell culture plate or dish, remove the cell culture medium and add the prepared Mito-Tracker Deep Red FM working solution. Incubate at 37°C for 15-30 minutes, then remove the Mito-Tracker Deep Red FM working solution and add fresh cell culture medium pre-incubated at 37°C. Observe using a fluorescence microscope. Mitochondria will be observed to exhibit bright, intense fluorescence.

[0291] Results: SZ-A could increase the number of mitochondria in endothelial cells and improve mitochondrial damage, indicating that SZ-A could significantly improve endothelial cell function (Figure 20).

[0292] Test Example 9: Clinical Evaluation

[0293] 1. Experimental Plan

[0294] 1.1 Experimental Grouping

[0295] This study adopted a prospective, single-arm, open-label, multicenter trial design. Inclusion criteria: (1) Males aged 45 to 70 years, married or unmarried with a regular sexual partner; (2) Subjects diagnosed with type 2 diabetes according to WHO criteria, with glycated hemoglobin ≥7%. (3) Clear consciousness and no cognitive abnormalities; (4) Signed informed consent. Exclusion criteria: (1) History of prostate, penis, or urethra surgery or trauma; (2) Congenital anomalies of the reproductive system; (3) Use of drugs that may cause ED in the past month; (4) Major physical diseases such as severe organ lesions and malignant tumors; (5) History of drug abuse; (6) Mental illness. All signed informed consent. A total of 17 subjects were enrolled.

[0296] 1.2 Administration

[0297] The experimental group took the tablets orally, with an initial dose of 1 tablet 3 times a day; after 4 weeks, the dose was increased to 2 tablets 3 times a day; the total treatment lasted 48 weeks.

[0298] Preparation method of tablets: Take the mulberry branch extract of Preparation Example 8, add appropriate amount of excipients, mix evenly, add water to make a soft material, granulate and dry, add magnesium stearate, mix evenly, and tablet to obtain a mulberry branch extract preparation, each tablet containing 50 mg of total alkaloids.

[0299] 1.3 Experimental data detection and processing

[0300] 1. At week 0 (before treatment) and week 48 (after treatment), the International Index of Erectile Function (IIEF) was used to assess the severity of ED in the subjects. The IIEF consists of 15 questions, all scored on a 6-point scale (0 to 5 points), with a total score of 75 points. The scoring table is as follows:

[0301] Table 5

[0302] Note: The International Index of Erectile Function (IIEF) was originally developed during the clinical development of sildenafil to more reliably assess erectile function and treatment efficacy in ED patients. Currently, the IIEF-15 has become the gold standard for evaluating the efficacy of ED in clinical trials and has been gradually applied to studies of different ED etiologies, comorbidities, cultural backgrounds, and treatment interventions. It consists of 15 items divided into five sections: erectile function (questions 1-5), satisfaction with penetration (questions 6-8), orgasm (questions 9-10), sexual desire (questions 11-12), and overall satisfaction (questions 13-15). Patients respond to each item based on their sexual activity over the past four weeks. Lower IIEF-15 scores indicate more severe ED (worse erectile function).

[0303] 2. Statistical Analysis

[0304] SPSS 20.0 software was used for data processing, and the differences before and after treatment were analyzed using T-test.

[0305] 3. Experimental Results

[0306] Results showed that after SZ-A treatment, IIEF-15 scores increased, suggesting that SZ-A can improve erectile function. SZ-A was found to improve five aspects: erectile function, satisfaction with penetration, orgasm, sexual desire, and overall satisfaction (Figure 21). Further analysis revealed that 11 of the 17 subjects experienced an increase in scores after SZ-A treatment, suggesting that SZ-A's treatment rate for ED is approximately 65%.

Claims

1. Use of mulberry extract in the preparation of a medicament for treating and / or preventing male infertility, characterized in that, Based on the total weight percentage of the components of the mulberry extract being 100%, the mulberry extract contains alkaloids with a weight content of more than 3% (optionally containing alkaloids with a weight content of 3 - 99%, further optionally containing alkaloids with a weight content of 15 - 99%, further optionally containing alkaloids with a weight content of 30 - 99%, further optionally containing alkaloids with a weight content of 40 - 99%, further optionally containing alkaloids with a weight content of 50 - 99%, further optionally containing alkaloids with a weight content of 60 - 99%). and / or contains polysaccharides with a weight content not higher than 70% (optionally containing polysaccharides with a weight content of 0.2 - 70%, further optionally containing polysaccharides with a weight content of 0.2 - 50%, further optionally containing polysaccharides with a weight content of 0.2 - 35%, further optionally containing polysaccharides with a weight content of 0.2 - 25%, further optionally containing polysaccharides with a weight content of 0.2 - 23%, further optionally containing polysaccharides with a weight content of 20 - 25%). and / or contains flavonoids with a weight content not higher than 10% (optionally containing flavonoids with a weight content of 0.05 - 5%, further optionally containing flavonoids with a weight content of 0 - 2%, further optionally containing flavonoids with a weight content of 0.05 - 2%, further optionally containing flavonoids with a weight content of 0.5 - 1.5%, further optionally containing flavonoids with a weight content of 0 - 1%, further optionally containing flavonoids with a weight content of 0.05 - 1%). and / or contains amino acids with a weight content not higher than 50% (optionally containing amino acids with a weight content of 0 - 30%, further optionally containing amino acids with a weight content of 0 - 25%, further optionally containing amino acids with a weight content of 0 - 20%, further optionally containing amino acids with a weight content of 0 - 5%, further optionally containing amino acids with a weight content of 3 - 25% or further optionally containing amino acids with a weight content of 5 - 20%). and / or other components (the weight content is optionally 0 - 25%, further optionally 0 - 20%, further optionally 0 - 15%, further optionally 0 - 11%, further optionally 2 - 20%, further optionally 4 - 8%).

2. The use according to claim 1, characterized in that, The male infertility is male sexual dysfunction and / or male reproductive dysfunction.

3. The use according to claim 1 or 2, characterized in that The male sexual dysfunction and / or male reproductive dysfunction is caused by endocrine disorders and / or metabolic disorders. Preferably, the metabolic disorder is abnormal glycolipid metabolism; preferably, the endocrine disorder is abnormal hormone levels. Preferably, the abnormal glycolipid metabolism is selected from at least one of diabetes, hyperglycemia, overweight, obesity, and hyperlipidemia.

4. The use according to any one of claims 1 to 3, characterized in that, The male sexual dysfunction is selected from at least one of sexual desire disorder, erectile dysfunction, ejaculation disorder, and anorgasmia; preferably, the male sexual dysfunction is erectile dysfunction and / or ejaculation disorder. The male reproductive dysfunction is selected from at least one of vas deferens obstruction, abnormal sperm quality, abnormal semen, varicocele and inflammation; further preferably, the abnormal sperm quality is selected from at least one of azoospermia, oligospermia, asthenospermia and asthenozoospermia; preferably, the male reproductive dysfunction is oligospermia, asthenospermia and / or oligoasthenospermia.

5. The use according to any one of claims 1 to 4, characterized in that, The treatment and / or prevention of male infertility is specifically manifested as at least one of the following: 1) Promote sperm production; 2) Improve sperm motility; 3) Improve sperm quality; 4) Improve testicular index; 5) Improve the structure of seminiferous tubules in testicular tissue and the morphology and quantity of spermatogenic cells; 6) Improve the levels of related sex hormones; preferably, the sex hormone is androgen; further preferably, the androgen is testosterone; 7) Improve the expression level of androgen synthase; 8) Improve the expression level of androgen receptor; 9) Improve penile tissue damage and / or cell damage; preferably, the penile cells are penile cavernosal endothelial cells; preferably, improve the content of NO in penile cavernosal endothelial cells; preferably, the improvement of penile tissue damage includes at least one of improving the average thickness of the cavernous scaffold, connection density, separation degree and the average intercept length of blood sinus cavities; Further preferably, the treatment and / or prevention of male reproductive dysfunction is specifically manifested as at least one of the following: 1) Promote sperm production; 2) Improve sperm motility; 3) Improve sperm quality; 4) Improve testicular index; 5) Improve the structure of seminiferous tubules in testicular tissue and the morphology and quantity of spermatogenic cells; 6) Improve the levels of related sex hormones; 7) Improve the expression level of androgen synthase; 8) Improve the expression level of androgen receptor; 9) Improve testicular cell damage; preferably, the testicular cells are Leydig cells; Further preferably, the treatment and / or prevention of male sexual dysfunction is specifically manifested as at least one of the following: 1) Improve the levels of related sex hormones; preferably, the sex hormone is androgen; further preferably, the androgen is testosterone; 2) Improve the expression level of androgen synthase; 3) Improve the expression level of androgen receptor; 4) Improve penile tissue and / or cell damage; preferably, the penile cells are penile cavernosal endothelial cells; preferably, improve the content of NO in penile cavernosal endothelial cells; preferably, the improvement of penile tissue damage includes at least one of improving the average thickness of the cavernous scaffold, connection density, separation degree and the average intercept length of blood sinus cavities; 5) Improve the IIEF-15 score; preferably, the IIEF-15 score includes at least one of erectile function, satisfaction with insertion, orgasm, sexual desire and overall satisfaction.

6. Use of mulberry extract in the preparation of a product for inhibiting apoptosis of Leydig cells induced by high glucose in vitro; Optionally, use of the mulberry extract in the preparation of a product for improving endothelial cell damage induced by high glucose in vitro, Based on the total weight percentage of the components of the mulberry extract being 100%, the mulberry extract contains alkaloids with a weight content of more than 3% (optionally containing alkaloids with a weight content of 3 - 99%, further optionally containing alkaloids with a weight content of 15 - 99%, further optionally containing alkaloids with a weight content of 30 - 99%, further optionally containing alkaloids with a weight content of 40 - 99%, further optionally containing alkaloids with a weight content of 50 - 99%, further optionally containing alkaloids with a weight content of 60 - 99%). and / or contains polysaccharides with a weight content not higher than 70% (optionally containing polysaccharides with a weight content of 0.2 - 70%, further optionally containing polysaccharides with a weight content of 0.2 - 50%, further optionally containing polysaccharides with a weight content of 0.2 - 35%, further optionally containing polysaccharides with a weight content of 0.2 - 25%, further optionally containing polysaccharides with a weight content of 0.2 - 23%, further optionally containing polysaccharides with a weight content of 20 - 25%). and / or contains flavonoids with a weight content not higher than 10% (optionally containing flavonoids with a weight content of 0.05 - 5%, further optionally containing flavonoids with a weight content of 0 - 2%, further optionally containing flavonoids with a weight content of 0.05 - 2%, further optionally containing flavonoids with a weight content of 0.5 - 1.5%, further optionally containing flavonoids with a weight content of 0 - 1%, further optionally containing flavonoids with a weight content of 0.05 - 1%). and / or contains amino acids with a weight content not higher than 50% (optionally containing amino acids with a weight content of 0 - 30%, further optionally containing amino acids with a weight content of 0 - 25%, further optionally containing amino acids with a weight content of 0 - 20%, further optionally containing amino acids with a weight content of 0 - 5%, further optionally containing amino acids with a weight content of 3 - 25% or further optionally containing amino acids with a weight content of 5 - 20%). and / or other components (the weight content is optionally 0 - 25%, further optionally 0 - 20%, further optionally 0 - 15%, further optionally 0 - 11%, further optionally 2 - 20%, further optionally 4 - 8%).

7. Use according to any one of claims 1-6, characterized in that, Based on the total weight percentage of the components of the mulberry extract being 100%, the weight content of each component in the mulberry extract is as follows: Optionally, based on the total weight percentage of the components of the mulberry extract being 100%, the weight content of each component in the mulberry extract is as follows: Optionally, based on the total weight percentage of the components of the mulberry extract being 100%, the weight content of each component in the mulberry extract is as follows: Optionally, based on the total weight percentage of the components of the mulberry extract being 100%, the weight content of each component in the mulberry extract is as follows: Optionally, based on the total weight percentage of the components of the mulberry extract being 100%, the weight content of each component in the mulberry extract is as follows: Optionally, based on the sum of the weight percentages of the components of the mulberry extract being 100%, the weight contents of the components in the mulberry extract are as follows: Optionally, based on the sum of the weight percentages of each component of the mulberry extract being 100%, the weight content of each component in the mulberry extract is as follows: Further optionally, based on the total weight percentage of the components of the mulberry extract being 100%, the weight content of each component in the mulberry extract is as follows: Further optionally, based on the total weight percentage of the components of the mulberry extract being 100%, the weight contents of the components in the mulberry extract are as follows: Further preferably, the alkaloid comprises one or more of 1-deoxynojirimycin (1-deoxynojirimycin or DNJ), N-methyl-1-deoxynojirimycin, fagomine (fagomine or FAG), 3-epi-fagomine, 1,4-dideoxy-1,4-imino-D-arabinitol (1,4-dideoxy-1,4-imino-D-arabinitol or DAB), calystegin B2, calystegin C1, 2-O-(α-D-galactopyranosyl)-1-deoxynojirimycin, 6-O-(β-D-glucopyranosyl)-1-deoxynojirimycin, 1,4-dideoxy-1,4-imino-(2-O-β-D-glucopyranosyl)-D-arabinitol; Optionally, the weight percentage of DNJ is not less than 50% of the total alkaloid (optionally 60-99%).

8. The use according to any one of claims 1-7, characterized in that The preparation method of the mulberry extract comprises the following steps: 1) Prepare a crude extract of Moraceae plants; 2) Separate the crude extract through a cation resin and / or an optional anion resin to obtain the mulberry extract; Preferably, the method further comprises the following steps: 3) Perform alcohol precipitation on the resin effluent of step 2), and collect the supernatant; 4) Concentrate and / or dry the supernatant; Optionally, it further comprises the following step: concentrate and / or dry the resin effluent of step 2).

9. The use according to any one of claims 1-8, characterized in that, The mulberry extract acts on humans or mammals.

10. The use according to any one of claims 1-9, characterized in that, The drug is in an oral dosage form; optionally, the drug is tablets, capsules, oral solutions, oral emulsions, pills, granules, syrups and powders.

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