Application of mulberry extract in preparation of medicine for treating and / or preventing diseases related to abnormal bone metabolism

Abstract

According to the application of the mulberry extract in preparing the medicine for treating and / or preventing the diseases related to the abnormal bone metabolism, the mulberry extract comprises alkaloid with the weight content being 3% or above, and / or polysaccharide with the weight content being not higher than 70%, and / or flavone with the weight content being not higher than 10%, and / or amino acid with the weight content being not higher than 50%, and / or other components. The mulberry extract disclosed by the invention has a remarkable improvement effect on diseases related to abnormal bone metabolism.

Description

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 diseases related to abnormal bone metabolism. Background Art

[0002] Bones are tissues that continuously perform remodeling metabolic activities, including two relative processes, namely bone formation (osteoblasts) and bone resorption (osteoclasts). Under normal circumstances, bone resorption and bone formation are closely related to different hormones (such as PTH, vitamin D, steroids and calcitonin, etc.) and intrinsic regulatory factors (such as cytokines and growth factors, etc.). The bone resorption process is regulated by RANKL and M-CSF and inhibited by OPG. Bone formation is induced by many biological factors; it is also regulated by M-CSF, ALP, osteocalcin, osteopontin and osteonectin. An imbalance between bone resorption and bone formation leads to bone metabolic diseases such as osteoporosis, osteoarthritis, rheumatoid arthritis and bone displacement.

[0003] Osteoporosis (OP) is a common metabolic bone disease characterized by decreased bone mass and destruction of bone tissue microstructure, which leads to increased bone brittleness, decreased bone strength, and susceptibility to fractures. The stability of bone metabolism is regulated by the balance between bone formation caused by osteoblasts and bone resorption caused by osteoclasts. Primary osteoporosis is the most common type in clinical practice, accounting for about 80%, and can be divided into type I high-turnover osteoporosis with increased bone resorption (common in postmenopausal women) and type II low-turnover osteoporosis with decreased bone formation (common in the elderly).

[0004] Sarcopenia is a clinical syndrome characterized by a progressive and widespread decrease in muscle mass and strength, which can lead to physical disability, decreased quality of life, and an increased risk of adverse consequences such as death. Sarcopenia and osteoporosis are closely related degenerative diseases, which are mainly manifested through the interaction between muscle and bone. A decrease in muscle mass will lead to and accelerate the occurrence and development of osteoporosis, while a decrease in bone strength will also promote atrophy of muscle morphology and functional degeneration.

[0005] There is currently no report on the use of mulberry extracts to treat diseases related to abnormal bone metabolism. Summary of the invention

[0006] The purpose of the present invention is to provide a use of a mulberry extract in preparing a drug for treating and / or preventing diseases related to abnormal bone metabolism. The mulberry extract of the present invention has a significant effect on improving diseases related to abnormal bone metabolism.

[0007] 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 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%, further optionally containing alkaloids with a weight content of 35-70%, further optionally containing alkaloids with a weight content of 60-75%;

[0008] and / or comprising a polysaccharide content of not more than 70% by weight (optionally comprising a polysaccharide content of 0.2-70% by weight, further optionally comprising a polysaccharide content of 0.2-50% by weight, further optionally comprising a polysaccharide content of 0.2-35% by weight, further optionally comprising a polysaccharide content of 0.2-25% by weight, further optionally comprising a polysaccharide content of 0.2-23% by weight, further optionally comprising a polysaccharide content of 20-25% by weight),

[0009] and / or contains flavonoids having a weight content of not more than 10% (optionally containing flavonoids having a weight content of 0.05-5%, further optionally containing flavonoids having a weight content of 0-2%, further optionally containing flavonoids having a weight content of 0.05-2%, further optionally containing flavonoids having a weight content of 0.5-1.5%, further optionally containing flavonoids having a weight content of 0-1%, further optionally containing flavonoids having a weight content of 0.05-1%),

[0010] and / or contains no more than 50% amino acids by weight (optionally contains 0-30% amino acids by weight, further optionally contains 0-25% amino acids by weight, further optionally contains 0-20% amino acids by weight, further optionally contains 0-5% amino acids by weight, further optionally contains 3-25% amino acids by weight or further optionally contains 5-20% amino acids by weight),

[0011] and / or other components (the weight content can be optionally 0-25%, further optionally 0-20%, further optionally 0-15%, further optionally 0-11%, further optionally 2-20%, further optionally 4-8%.

[0012] Optionally, the disease related to abnormal bone metabolism is any one of osteoporosis, sarcopenia, sarcopenia-osteoporosis, muscular dystrophy, myasthenia, osteomalacia, rickets, primary hyperparathyroidism bone disease, fluorosis, osteoarthritis and bone displacement; it can be osteoporosis, sarcopenia or sarcopenia-osteoporosis.

[0013] In addition, diseases related to abnormal bone metabolism may also include kidney disease, liver disease, diabetes, rheumatoid arthritis and other diseases that cause decreased bone density.

[0014] Optionally, the use of the mulberry extract in the preparation of a medicament for treating and / or preventing diseases related to abnormal bone metabolism includes: the use of the mulberry extract in any one of the following (b1)-(b3):

[0015] (b1) Use in the preparation of a drug for increasing bone density;

[0016] (b2) Use in the preparation of a drug for reducing bone marrow fat cavitation;

[0017] (b3) Use in the preparation of drugs for improving bone microstructure;

[0018] The improvement of bone microstructure includes at least one of the following: (i) increasing the number of trabeculae; (ii) reducing the separation of trabeculae; (iii) reducing the trabecular structure model index; (iv) increasing the bone volume fraction;

[0019] Optionally, the use of the mulberry extract in the preparation of a medicament for treating and / or preventing diseases related to abnormal bone metabolism includes: the use of the mulberry extract in any of the following:

[0020] 1) Use in the preparation of drugs for increasing muscle mass;

[0021] 2) Use in the preparation of drugs for increasing muscle weight percentage;

[0022] 3) Use in the preparation of drugs for increasing muscle fat ratio;

[0023] 4) Use in the preparation of a drug for inhibiting muscle atrophy and / or edema; optionally, the inhibiting muscle atrophy and / or edema is specifically to increase the cross-sectional area, muscle fiber diameter and / or muscle fiber circumference of the muscle fiber;

[0024] 5) Use in the preparation of drugs for relieving intermuscular peripheral neuropathy and / or vascular lipid deposition;

[0025] 6) Use in the preparation of drugs for inhibiting muscle cell damage;

[0026] 7) Use in the preparation of drugs for increasing muscle strength;

[0027] In an optional embodiment of the present invention, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0028]

[0029] Optionally, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0030]

[0031]

[0032] Optionally, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0033]

[0034] Optionally, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0035]

[0036] Optionally, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0037]

[0038] Optionally, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0039]

[0040] Optionally, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0041]

[0042] Further optionally, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0043]

[0044] Further optionally, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0045]

[0046] Further optionally, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0047]

[0048] Optionally, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0049]

[0050] Optionally, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0051]

[0052] Optionally, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0053]

[0054] Optionally, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0055]

[0056] Optionally, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0057]

[0058] Optionally, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0059]

[0060] Optionally, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0061]

[0062] Optionally, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0063]

[0064] Optionally, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0065]

[0066]

[0067] Optionally, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0068]

[0069] Optionally, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0070]

[0071] Optionally, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0072]

[0073] Optionally, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0074]

[0075]

[0076] Optionally, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0077]

[0078] Optionally, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0079]

[0080] Optionally, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0081]

[0082] Optionally, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0083]

[0084]

[0085] Optionally, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0086]

[0087] Optionally, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0088]

[0089] Optionally, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0090]

[0091] Optionally, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0092]

[0093]

[0094] Optionally, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0095]

[0096] Optionally, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0097]

[0098] Optionally, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0099]

[0100] Optionally, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0101]

[0102] Optionally, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0103]

[0104] Optionally, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0105]

[0106] Optionally, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0107]

[0108] Optionally, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0109]

[0110] Optionally, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0111]

[0112] Optionally, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

[0113]

[0114] The weight content of each component can be any combination of the above values.

[0115] Optionally, the alkaloids include 1-deoxynojirimycin (DNJ), N-methly-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-oxo-(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.

[0116] Wherein, optionally, the weight percentage of DNJ is not less than 50% (optionally 60-99%) of the total alkaloids.

[0117] Optionally, the heavy metal content of the mulberry extract does not exceed 10 ppm.

[0118] The drug is in the form of an oral administration 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.

[0119] In the present invention, the mulberry extract can be provided in the form of commercially available mulberry branch total alkaloid tablets (National Medicine Standard No. Z20200002).

[0120] Another aspect of the present invention provides use of the mulberry extract in treating and / or preventing diseases related to abnormal bone metabolism.

[0121] Optionally, the use of the mulberry extract in treating and / or preventing diseases related to abnormal bone metabolism includes: the use of the mulberry extract in any one of the following (b1)-(b3):

[0122] (b1) Use in increasing bone density;

[0123] (b2) Use in reducing bone marrow fat vacuoles;

[0124] (b3) Use in improving bone microstructure;

[0125] The improvement of bone microstructure includes at least one of the following: (i) increasing the number of trabeculae; (ii) reducing the separation of trabeculae; (iii) reducing the trabecular structure model index; (iv) increasing the bone volume fraction;

[0126] Optionally, the use of the mulberry extract in treating and / or preventing diseases related to abnormal bone metabolism includes: the use of the mulberry extract in any of the following:

[0127] 1) Use in increasing muscle mass;

[0128] 2) Use in increasing muscle mass percentage;

[0129] 3) Use in increasing muscle-to-fat ratio;

[0130] 4) Use in inhibiting muscle atrophy and / or edema; optionally, the inhibiting muscle atrophy and / or edema is specifically to increase the cross-sectional area of ​​muscle fibers, the diameter of muscle fibers and / or the circumference of muscle fibers;

[0131] 5) Use in alleviating intermuscular peripheral neuropathy and / or vascular lipid deposition;

[0132] 6) Use in inhibiting muscle cell damage;

[0133] 7) Use in increasing muscle strength;

[0134] Preferably, the bones include tibia and femur.

[0135] The muscles include skeletal muscles, such as gastrocnemius, tibialis anterior, quadriceps femoris and soleus, etc. The mulberry extract acts on humans or mammals.

[0136] Optionally, the treatment and / or prevention of diseases related to abnormal bone metabolism is reflected in improving any one of the patient's bone mass, bone quality, muscle mass, muscle strength, and body balance; optionally, the patient suffers from any one of osteoporosis, sarcopenia, sarcopenia-osteoporosis (muscular dystrophy), muscular dystrophy, myasthenia, osteomalacia, rickets, primary hyperparathyroidism bone disease, fluorosis, osteoarthritis and bone dislocation; further optionally, the patient suffers from osteoporosis, sarcopenia or sarcopenia-osteoporosis.

[0137] Optionally, the disease related to abnormal bone metabolism is caused by aging, endocrine disorder, obesity and / or diabetes.

[0138] Optionally, 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 through 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.

[0139] 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; 4) concentrating and / or drying the supernatant. Optionally, before subjecting the resin effluent of step 2) to alcohol precipitation, it is concentrated and / or dried.

[0140] Optionally, the moraceae plant 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, and a combination of one or more of the above mulberry species bred intraspecifically or interspecifically;

[0141] Optionally, the moraceae plant is one or more selected from 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 Sangte You No. 2. Leaves, roots, branches, bark, buds, stems, fruits and other parts of the moraceae plant can be used, and mulberry branches, mulberry leaves or white mulberry bark can be used.

[0142] 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.

[0143] In one embodiment, the mulberry extract is prepared according to the following steps: crushing the moraceae plant, heating and refluxing extraction 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 part, adding ethanol, precipitating to remove impurities, concentrating and / or drying to obtain an extract.

[0144] In one embodiment, the mulberry extract is prepared according to the following steps: crushing the moraceae plant, heating and refluxing extraction 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 part, concentrating and / or drying to obtain an extract.

[0145] In one embodiment, the mulberry extract is prepared according to the following steps: crushing the moraceae plant, heating and refluxing extraction 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 an extract.

[0146] In one embodiment, the mulberry extract is prepared according to the following steps: crushing the moraceae plant, heating and refluxing extraction 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 0.5-3h each time, further optionally 1-2h 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 part (i.e., the anion resin effluent), adding ethanol, precipitating to remove impurities, concentrating and / or drying to obtain an extract.

[0147] 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 period of 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, and 0.4-10 times can be selected 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, and further optionally 80-400rpm or 300rpm.

[0148] 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, and can be 8.5-9.5; Optionally, the alkaline solution is selected from an aqueous ammonia solution, a sodium hydroxide solution, a potassium hydroxide solution, or a sodium carbonate solution; Optionally, the concentration of the alkaline solution is 0.5-4 mol / L, and can be 1-2 mol / L. Optionally, the acidic solution is washed until the pH of the eluate is 3.0-7.0, and can be 4.5-6.5. Optionally, the acidic solution is selected from a hydrochloric acid solution, a phosphoric acid solution, and a sodium hydrogen phosphate-citric acid buffer solution, and Optionally, the concentration of the acidic solution is 0.5-4 mol / L, and can be 1.5-2 mol / L. Optionally, the cationic resin can also be rinsed with 3-5 column volumes of deionized water after the last acidic solution wash.

[0149] 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.

[0150] 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).

[0151] After the plant crude 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.

[0152] 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).

[0153] Optionally, the eluent flow rate is 1-15 BV / h (optionally 5-10 BV / h, further optionally 5-6 BV / h).

[0154] Optionally, the weight of the eluent used for cationic resin separation is 0.1-30 times the weight of the plant raw material feed. Optionally, the eluent is 0.5-10 times the weight of the plant raw material feed for elution.

[0155] The collection starting point can be determined according to the pH of the effluent from the cationic resin. For example, when an alkaline solution such as aqueous ammonia is used for elution, the eluate is collected when the pH of the effluent from the cationic column is detected to be greater than 7, or the collection starting point of the effluent is determined according to 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 input, the collection is stopped, and the collected liquid is optionally purified by an anion column.

[0156] When purifying by anion column, optionally, after the anion resin is loaded into the column, it is activated in the order of washing with alkaline solution, washing with acidic solution, and washing with alkaline solution.

[0157] Optionally, washing with an alkaline solution until the pH of the washing solution is 8.0-9.5, optionally 8.5-9.5;

[0158] 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.

[0159] 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 the concentration of the acidic solution is 0.5-4 mol / L, optionally 1-2 mol / L.

[0160] 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.

[0161] 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).

[0162] When the liquid flows out of the anion resin, the collection begins. Optionally, 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, the collection is stopped.

[0163] 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 10-600rpm (optionally 40-500rpm, 80-400rpm). The alcohol precipitation treatment time is 12-24h.

[0164] Furthermore, before the alcohol precipitation treatment, the anion resin effluent is centrifuged or filtered through a microfiltration membrane to remove impurities, and then concentrated through a reverse ion osmosis membrane. The specific gravity of the concentrated liquid can be 1.0-1.3, and can be optionally 1.1-1.25.

[0165] Optionally, the drug further includes a pharmaceutically acceptable carrier. The carrier is an inactive ingredient that is non-toxic to the human body and conforms to the route of administration or the mode of administration. The carrier can be a solid or liquid excipient. Solid excipients, for example, include 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, polyvinylpyrrolidone, starch, magnesium stearate, sodium carboxymethyl starch and talc; liquid excipients, for example, include water, ethanol, syrup and glycerol.

[0166] In the present invention, the mulberry extract is referred to as SZ-A.

[0167] Beneficial Effects

[0168] Mulberry extract can increase bone density, promote osteoblast bone formation, inhibit osteoclast bone resorption, and at the same time improve muscle content and quality and promote myoblast proliferation and differentiation.

[0169] The present invention provides the use of mulberry extract in preparing medicines or health products for treating and / or preventing diseases with abnormal bone metabolism, which can significantly improve the bone density, bone calcium content, muscle content, etc. of model mice, and ultimately improve diseases related to abnormal bone metabolism. BRIEF DESCRIPTION OF THE DRAWINGS

[0170] Figure 1These are the results of bone density, muscle weight percentage, and muscle weight / fat weight determination of mice in each group of Experimental Example 1, where: *P<0.05, **P<0.01, ***P<0.001 vs. model group.

[0171] Figure 2 The graph of pathological sections of gastrocnemius muscle of each group of mice in Experimental Example 1 (2A), the bar graph of average cross-section of gastrocnemius muscle fibers of each group of mice (2B), the bar graph of average diameter of gastrocnemius muscle fibers (2C), and the bar graph of average circumference of gastrocnemius muscle fibers (2D); the graph of pathological sections of tibia of mice ( Figure 2 E).

[0172] Figure 3 3A is the microCT scanning result of bone tissue of mice in each group of Experimental Example 2, and 3B is the trabeculae, 3C is the trabecular separation, 3D is the bone density, 3E is the bone volume fraction, 3F is the bone structure model index, 3G is the trabecular thickness and other results of the bar graph, among which *P<0.05, **P<0.01 vs. the model group.

[0173] Figure 4 These are the results of bone density, muscle weight percentage, and muscle weight / fat weight determination of mice in each group of Experimental Example 2, where: *P<0.05, **P<0.01, ***P<0.001, vs. model group.

[0174] Figure 5 The results of bone density measurement of mice in each group in Experimental Example 2, where *P<0.05, **P<0.01, vs. model group.

[0175] Figure 6 These are photos of femoral pathological sections of mice in each group in Experimental Example 3.

[0176] Figure 7 These are the results of Experimental Example 4, the effect of mulberry extract on muscle weight percentage in LZ+HFD-induced endocrine disorder model rats, *P<0.05, **P<0.01, vs. model group.

[0177] Figure 8 A in the middle is the effect of mulberry extract in Experimental Example 5 on the pathological sections of the gastrocnemius muscle in the Apoe mouse diabetic sexual dysfunction model induced by TZ combined with high-fat and high-cholesterol; BD is the bar graph of the effect of mulberry extract in Experimental Example 5 on the average area, average diameter and average circumference of the gastrocnemius muscle fibers in the Apoe mouse diabetic sexual dysfunction model induced by TZ combined with high-fat and high-cholesterol, *P<0.05, **P<0.01, vs. model group.

[0178] Fig. 9It is a bar graph of the bone density measurement results of mice in each group of Experimental Example 6, *P<0.05, **P<0.01, ***P<0.001, vs. aging group.

[0179] Fig.10 It is a bar graph showing the grip strength results of each group of mice in Experimental Example 7 (*P<0.05 vs. model group; ###P<0.001 vs. normal group).

[0180] Fig.11 Shown are SA-β-galactosidase staining images (A) and statistical bar graphs (B) of the effects of mulberry extracts prepared in different examples in Experimental Example 8 on D-galactose modeled C2C12 cells, ***P<0.001, ****P<0.0001, vs. Model group.

[0181] Fig.12 These are histograms of (A) DNJ, (B) FA, and (C) DAB distribution in bone marrow tissue of rats of Experimental Example 9 after oral administration of SZ-A (40 mk / kg). DETAILED DESCRIPTION

[0182] The present invention is further described below through specific experimental examples. It should be understood that the following experiments are only used to illustrate the technical solution of the present invention and are not used to limit the protection scope of the present invention.

[0183] The mulberry extract of the present invention can be prepared by the following method:

[0184] 1. Example of Preparation of Mulberry Extract

[0185] Preparation Example 1 of Mulberry Extract

[0186] Take 1000kg of fresh mulberry branches (fine-toothed mulberry branches, Yuesang 11), crush them, add 4000L of water, extract them by heating reflux for 2h, combine the extracts, filter and remove insoluble matter to obtain a crude extract. The crude extract is heat concentrated to a solid mass percentage of 4%, and kept at 50℃ as the loading liquid for the cationic resin column.

[0187] Use 150kg of D113 macroporous weakly acidic phenyl propylene cationic resin to load the column, wash with 2mol / L hydrochloric acid solution until the pH of the eluate is 4.5; wash with 1mol / L sodium hydroxide solution until the pH of the eluate is 8.5; wash with 2mol / L hydrochloric acid solution until the pH of the eluate is 4.5; then rinse with 5 times the column volume of deionized water to complete the activation. Load the concentrated extract, then elute with 1000L 2.5mol / L ammonia water at a rate of 6BV / h. Collect the eluate when the pH of the cation column effluent is >7. When the collected liquid reaches 900L, stop collecting and directly purify the collected liquid through the anion column.

[0188] Use 62.5 kg of D218 macroporous strongly alkaline acrylic anion resin to load the column, wash with 1.5 mol / L sodium hydroxide solution until the pH of the eluate is 9.0; wash with 1.5 mol / L hydrochloric acid solution until the pH of the eluate is 3.5; wash with 1.5 mol / L sodium hydroxide solution until the pH of the eluate is 9.0; complete activation. Load the collected cationic resin eluate onto the anion resin, and collect the effluent until the effluent reaches 920 L.

[0189] The collected liquid was centrifuged to remove impurities and then concentrated by reverse ion osmosis membrane. The liquid after concentration had a specific gravity of 1.25, and was transferred to an alcohol precipitation tank. 25L of anhydrous ethanol was added at 500rpm. After the addition of ethanol, stirring was stopped, and the alcohol precipitation was carried out for 24 hours. The supernatant was taken and concentrated under reduced pressure to obtain a mulberry branch extract (i.e., mulberry extract SZ-A).

[0190] The mulberry branch extract has 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%.

[0191] Preparation Example 2 of Mulberry Extract

[0192] Take 10kg of fresh mulberry branches (Sangteyou No. 2), crush them, add 150L of water, add them in 2 times, extract them by decoction for 3h each time, combine the extracts, and filter to remove insoluble matter. The extract is hot concentrated to 8% solid mass percentage, transferred to an alcohol precipitation tank, and 2367.9g of anhydrous ethanol (3L) is added under a stirring paddle of 300rpm. After the addition of ethanol, stop stirring, precipitate with alcohol for 24h, and take the supernatant as the sample liquid for the cationic resin column. Use 002SC type strong acid styrene cationic resin 5kg to load the column, and activate the cationic resin according to the method of Preparation Example 1. Load the extract treated with concentrated alcohol precipitation, then use 100L 5mol / L potassium chloride for elution, the elution rate is 5BV / h, and detect the effluent with 20% silicotungstic acid. Start collecting when a white precipitate is generated, and stop collecting when the collected liquid reaches 25L, and directly purify the collected liquid through an anion column.

[0193] Use 10 kg of 711 type strong basic styrene anion resin to load the column, and activate the anion resin according to the method of Preparation Example 1. Load the collected cationic resin eluate onto the anion resin, and collect the effluent until the effluent reaches 15 L. Load the collected liquid onto the cationic resin again, and separate twice more using the cationic resin and the anionic resin in turn according to the above method.

[0194] The collected liquid obtained after three column separations was centrifuged to remove impurities and then concentrated by reverse ion permeation 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 of 1000 rpm. After the addition of ethanol, stirring was stopped, and the alcohol was precipitated for 24 hours. The supernatant was taken and concentrated under reduced pressure to obtain an extract. In addition, fresh mulberry bark and mulberry leaf (Sangteyou No. 2) were extracted, and the extraction method and parameters were the same as the above method.

[0195] The obtained mulberry branch extract has a mass percentage of alkaloids of 98%, a mass percentage of polysaccharides of 0.2%, a mass percentage of flavonoids of 0.05%, and a mass percentage of amino acids of 0. Among the alkaloids, the content of 1-DNJ is 99%, FAG is 0.5%, and DAB is 0.4%.

[0196] The obtained Morus alba extract has 95% by weight of alkaloids, 2% by weight of polysaccharides, 0.1% by weight of flavonoids, and 1% by weight of amino acids. Among the alkaloids, the content of 1-DNJ is 96%, FAG is 1.5%, and DAB is 1.4%.

[0197] The obtained mulberry leaf extract has 90% by weight of alkaloids, 4% by weight of polysaccharides, 0.1% by weight of flavonoids, and 3% by weight of amino acids. Among the alkaloids, the content of 1-DNJ is 91%, FAG is 3.1%, and DAB is 2.8%.

[0198] Preparation Example 3 of Mulberry Extract

[0199] Take 1000kg of fresh mulberry branches (Guangdong mulberry), crush them, add 11500L of water, heat and reflux for 2h, combine the extracts, filter and remove insoluble matter to obtain a crude extract. The crude extract is first centrifuged to remove impurities, and then concentrated with a counter ion permeation membrane to a solid mass percentage of 1%, which is used as the loading solution for the cationic resin column.

[0200] 300 kg of D001 macroporous strong acid styrene cationic resin was loaded into the column, and the cationic resin 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 detected with 20% silicotungstic acid. When a white precipitate was generated, collection began, and when the collected liquid reached 1000 L, collection was stopped.

[0201] The collected liquid obtained after the cationic column separation is concentrated by nanofiltration membrane, and the extract is concentrated under reduced pressure to obtain an extract concentrate.

[0202] The obtained mulberry branch extract has 15% by weight of alkaloids, 20% by weight of polysaccharides, 7% by weight of flavonoids, and 45% by weight of amino acids. Among the alkaloids, the content of 1-DNJ is 55%, FAG is 23%, and DAB is 10%.

[0203] Preparation Example 4 of Mulberry Extract

[0204] 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.

[0205] Use 150kg of D113 macroporous weakly acidic phenyl propylene cationic resin to load the column, wash with 2mol / L hydrochloric acid solution until the pH of the eluate is 4.5; wash with 1mol / L sodium hydroxide solution until the pH of the eluate is 8.5; wash with 2mol / L hydrochloric acid solution until the pH of the eluate is 4.5; then rinse with 5 times the column volume of deionized water to complete the activation. Load the concentrated extract, then elute with 1000L 2.5mol / L ammonia water at a rate of 6BV / h. Collect the eluate when the pH of the cation column effluent is >7. When the collected liquid reaches 900L, stop collecting and directly purify the collected liquid through the anion column.

[0206] Use 125kg of D218 macroporous strongly alkaline acrylic anion resin to load the column, wash with 1.5mol / L sodium hydroxide solution until the pH of the eluate is 9.0; wash with 1.5mol / L hydrochloric acid solution until the pH of the eluate is 3.5; wash with 1.5mol / L sodium hydroxide solution until the pH of the eluate is 9.0; complete activation. Load the collected cationic resin eluate onto the anion resin, and collect the effluent with a pH greater than 8 until the effluent reaches 870L.

[0207] The collected liquid obtained after the anion column separation is filtered through a microfiltration membrane to remove impurities and then concentrated with a counter ion permeation membrane. The concentrated liquid has a specific gravity of 1.1 and is transferred to an alcohol precipitation tank. 15 kg of anhydrous ethanol is added at a stirring paddle of 400 rpm. After the addition of ethanol, stirring is stopped, and the alcohol precipitation is carried out for 24 hours. The supernatant is taken and concentrated under reduced pressure to obtain a mulberry branch extract. Sample content: the mass percentage of alkaloids is 80%, the mass percentage of polysaccharides is 5%, the mass percentage of flavonoids is 0.1%, and the mass percentage of amino acids is 4%. Among the alkaloids, the content of 1-DNJ is 75%, FAG is 12%, and DAB is 10%.

[0208] Preparation Example 5 of Mulberry Extract

[0209] 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.

[0210] Use 62.5 kg of D218 macroporous strongly alkaline acrylic anion resin to load the column, wash with 1.5 mol / L sodium hydroxide solution until the pH of the eluate is 9.0; wash with 1.5 mol / L hydrochloric acid solution until the pH of the eluate is 3.5; wash with 1.5 mol / L sodium hydroxide solution until the pH of the eluate is 9.0; complete activation. Load the collected extract concentrate onto the anion resin and collect the effluent.

[0211] The collected solution obtained after the anion column separation was filtered through a microfiltration membrane to remove impurities and then concentrated by a counter ion permeation membrane, and further concentrated and dried under reduced pressure to obtain a mulberry branch extract. Sample content: 3% by mass of alkaloids, 70% by mass of polysaccharides, 10% by mass of flavonoids, and 10% by mass of amino acids. Among the alkaloids, the content of 1-DNJ was 68%, FAG was 17%, and DAB was 8%.

[0212] Preparation Example 6 of Mulberry Extract

[0213] Take 1500kg of fresh mulberry branches (fine-toothed mulberry branches, Yuesang 11), crush them, add 6000L of water, extract them by heating reflux for 2h, combine the extracts, filter and remove insoluble matter to obtain a crude extract. The crude extract is heat concentrated to a solid mass percentage of 4%, and kept at 50℃ as the loading liquid for the cationic resin column.

[0214] Use 100kg of D113 macroporous weakly acidic phenyl propylene cationic resin to load the column, wash with 2mol / L hydrochloric acid solution until the pH of the eluate is 4.5; wash with 1mol / L sodium hydroxide solution until the pH of the eluate is 8.5; wash with 2mol / L hydrochloric acid solution until the pH of the eluate is 4.5; then rinse with 5 times the column volume of deionized water to complete the activation. Load the concentrated extract, then elute with 1000L 2.5mol / L ammonia water at a rate of 6BV / h. Collect the eluate when the pH of the cation column effluent is >7. Stop collecting when the collected liquid reaches 900L, and directly purify the collected liquid through the anion column.

[0215] Use 62.5kg of D218 macroporous strong alkaline acrylic anion resin to load the column, use 1.5mol / L sodium hydroxide solution to wash until the pH of the eluate is 9.0; use 1.5mol / L hydrochloric acid solution to wash until the pH of the eluate is 3.5; use 1.5mol / L sodium hydroxide solution to wash until the pH of the eluate is 9.0; complete activation. Load the collected cationic resin eluate onto the anion resin, and collect the effluent until the effluent reaches 870L. Concentrate the effluent under reduced pressure to obtain a mulberry branch extract, in which the mass percentage of alkaloids is 30%, the mass percentage of polysaccharides is 35%, the mass percentage of flavonoids is 2%, and the mass percentage of amino acids is 25%. Among the alkaloids, the content of 1-DNJ is 62%, FAG is 20%, and DAB is 13%.

[0216] Preparation Example 7 of Mulberry Extract

[0217] Take 1000kg of fresh mulberry branches (fine-toothed mulberry branches, Yuesang 11), crush them, add 4000L of water, extract them by heating reflux for 2h, combine the extracts, filter and remove insoluble matter to obtain a crude extract. The crude extract is heat concentrated to a solid mass percentage of 4%, and kept at 50℃ as the loading liquid for the cationic resin column.

[0218] Use 100kg of D113 macroporous weakly acidic phenyl propylene cationic resin to load the column, wash with 2mol / L hydrochloric acid solution until the pH of the eluate is 4.5; wash with 1mol / L sodium hydroxide solution until the pH of the eluate is 8.5; wash with 2mol / L hydrochloric acid solution until the pH of the eluate is 4.5; then rinse with 5 times the column volume of deionized water to complete the activation. Load the concentrated extract, then elute with 1000L 2.5mol / L ammonia water at a rate of 6BV / h. Collect the eluate when the pH of the cation column effluent is >7. Stop collecting when the collected liquid reaches 900L, and directly purify the collected liquid through the anion column.

[0219] Use 62.5kg of D218 macroporous strong alkaline acrylic anion resin to load the column, use 1.5mol / L sodium hydroxide solution to wash until the pH of the eluate is 9.0; use 1.5mol / L hydrochloric acid solution to wash until the pH of the eluate is 3.5; wash with 1.5mol / L sodium hydroxide solution until the pH of the eluate is 9.0; complete activation. Load the collected cationic resin eluate onto the anion resin, and collect the effluent until the effluent reaches 870L. Concentrate the effluent under reduced pressure to obtain a mulberry branch extract, in which the mass percentage of alkaloids is 40%, the mass percentage of polysaccharides is 25%, the mass percentage of flavonoids is 0.5%, and the mass percentage of amino acids is 25%. Among the alkaloids, the content of 1-DNJ is 57%, FAG is 24%, and DAB is 16%.

[0220] Preparation Example 8 of Mulberry Extract

[0221] 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.

[0222] Use 150kg of D113 macroporous weakly acidic phenyl propylene cationic resin to load the column, wash with 2mol / L hydrochloric acid solution until the pH of the eluate is 4.5; wash with 1mol / L sodium hydroxide solution until the pH of the eluate is 8.5; wash with 2mol / L hydrochloric acid solution until the pH of the eluate is 4.5; then rinse with 5 times the column volume of deionized water to complete the activation. Load the concentrated extract, then elute with 1000L 2.5mol / L ammonia water at a rate of 6BV / h. Collect the eluate when the pH of the cation column effluent is >7. When the collected liquid reaches 900L, stop collecting and directly purify the collected liquid through the anion column.

[0223] Use 62.5kg of D218 macroporous strongly alkaline acrylic anion resin to load the column, wash with 1.5mol / L sodium hydroxide solution until the pH of the eluate is 9.0; wash with 1.5mol / L hydrochloric acid solution until the pH of the eluate is 3.5; wash with 1.5mol / L sodium hydroxide solution until the pH of the eluate is 9.0; complete activation. Load the collected cationic resin eluate onto the anion resin, and collect the effluent with a pH greater than 8 until the effluent reaches 870L.

[0224] The collected liquid obtained after the anion column separation is filtered through a microfiltration membrane to remove impurities and then concentrated with a counter ion permeation membrane. The concentrated liquid has a specific gravity of 1.1 and is transferred to an alcohol precipitation tank. 15 kg of anhydrous ethanol is added at a stirring paddle of 400 rpm. After the addition of ethanol, stirring is stopped, and the alcohol precipitation is carried out for 24 hours. The supernatant is taken and concentrated under reduced pressure to obtain a mulberry branch extract. Sample content: the mass percentage of alkaloids is 63%, the mass percentage of polysaccharides is 23%, the mass percentage of flavonoids is 1%, and the mass percentage of amino acids is 5%.

[0225] Among the alkaloids, the content of 1-DNJ was 61.9%, FAG was 16.6%, and DAB was 11.1%.

[0226] 2. Efficacy test of mulberry extract

[0227] Experimental Example 1: High-fat diet (HFD) mouse model

[0228] Forty-five healthy male C57 mice aged 6 weeks were randomly divided into a normal group (NC), a high-fat model group (HFD), and different dose groups of mulberry extract (SZ-A) (divided into a low-dose group SZ-A100 (100 mg / kg / d, calculated as alkaloids), a medium-dose group SZ-A200 (200 mg / kg / d, calculated as alkaloids), and a high-dose group SZ-A300 (300 mg / kg / d, calculated as alkaloids)), with 9 mice in each group. The mulberry extract was the mulberry extract prepared in Preparation Example 8. 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, 60 kcal% Fat); after feeding for 17 weeks, each experimental group was treated with the corresponding dose of the test drug, and the drug was administered 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. The effect of SZ-A was evaluated by testing bone density and other indicators before, 4 weeks after, and 6 weeks after administration. Body fat content, muscle content, etc. were tested before, 4 weeks after, and 6 weeks after the experiment. The experimental mice were anesthetized with Shutai and placed prone in a FAXITRON ULTRAFOCUS100 bone density analyzer. The mice were scanned by X-rays to detect and analyze the fat, muscle content, and bone density in the mice. After the mice were sampled, the tibia and gastrocnemius were collected for HE staining. The femur was collected and fixed in 10% neutral buffered formalin for 24 hours, and excess soft tissue was removed. The femur was scanned using the Skyscan 1276micro-CT instrument using the following settings, and the image was reconstructed using NRecon software.

[0229] (1) The results of mouse bone density, muscle content to body weight ratio, muscle weight percentage, and muscle fat ratio are as follows Figure 1 The results showed that the bone density of mice in the model group was significantly reduced, and after 6 weeks of treatment, the bone density of mice in each dose group of SZ-A was significantly increased, indicating that mulberry extract can significantly increase bone density; the muscle weight percentage and muscle fat ratio of mice in the model group were significantly reduced, and after 6 weeks of treatment, each dose group of SZ-A could significantly increase the muscle weight percentage and muscle fat ratio of obese mice.

[0230] (2) Pathological examination

[0231] HE staining was used to observe the morphological changes of muscle fibers. Figure 2A is a pathological section of the gastrocnemius muscle of mice: the skeletal muscle structure of the normal group is clear, and no abnormalities are found in the muscle bundles and intermuscular fiber spaces, and no abnormalities are found in the intermuscular nerves and blood vessels. In the HFD group, a large number of muscle fibers were edematous, and muscle fiber edema and atrophy coexisted, with edema being the main factor; the cross-sections of muscle fibers in the same muscle bundle were significantly different and varied in size. Lipid deposits were seen under the intima of the arterioles in the intermuscular space, and the media of the arteries was slightly thickened. It can be seen that the density of myelin fibers in the intermuscular peripheral nerves was reduced (demyelinating changes). Compared with the model group, after the administration of SZ-A300, the muscle fiber structure was clear, edema and muscle fiber atrophy were alleviated, the arteriolar structure was clear, no obvious lipid deposition was observed, and peripheral neuropathy was alleviated.

[0232] Figure 2 B-2D results showed that compared with the model group, the average cross-section, average diameter and average circumference of gastrocnemius muscle fibers increased significantly after SZ-A300 administration.

[0233] Tibia was stained with H&E. Pathological results showed that SZ-A administration could reduce the number of fat vacuoles in bone tissue. Figure 2 E.

[0234] (3) MicroCT scanning of bone tissue, the results are as follows Figure 3 As shown, the number of trabeculae and bone density in the model group were significantly reduced, and the separation of trabeculae increased. SZ-A300 can improve the femoral microstructure of HFD mice, increase the number of trabeculae and bone density, and reduce the separation of trabeculae. The bone volume fraction in the model group was significantly reduced, and the trabecular structure model index increased. SZ-A has a trend of increasing the bone volume fraction and reducing the trabecular structure model index.

[0235] Experimental Example 2: GAN diet feeding mouse model

[0236] The GAN diet is a modified, trans-fat-free, high-fat, high-cholesterol, high-fructose diet (containing 40% fat, 20% fructose, and 2% cholesterol).

[0237] 6-week-old healthy male C57BL / 6J mice were selected and divided into a normal group, a model group, and a mulberry extract (SZ-A) group. The mulberry extract components were divided into a low-dose group SZ-A100 (100 mg / kg / d, calculated as alkaloids), a medium-dose group SZ-A200 (200 mg / kg / d, calculated as alkaloids), and a high-dose group SZ-A300 (300 mg / kg / d, calculated as alkaloids). The mulberry extract was prepared in Example 8. Among them, the normal group mice were fed with a normal feed group (normal saline), and the model group and the SZ-A group were given GAN diet (Research Diet, D09100310). An animal model was established. After 20 weeks of induction, the drug was continuously administered for six weeks. The mulberry extract group was given drugs, and the normal feed group and the model group were given normal saline. The corresponding feed was continued to be fed during the treatment. The specific animal grouping and drug treatment are shown in Table 1 below. Body fat content was detected after 4 weeks and 6 weeks of administration. The experimental mice were anesthetized with Zotacil and placed in a prone position in a FAXITRON ULTRAFOCUS100 bone density analyzer. The mice were scanned by X-rays to detect and analyze the body fat, muscle content, fat percentage and bone density.

[0238] Table 1 Animal groups and drug treatments

[0239]

[0240] Figure 4 The results showed that the bone density of mice in the model group was significantly reduced, and after 6 weeks of treatment, the bone density of mice in the SZ-A groups increased significantly, indicating that mulberry extract can significantly increase the bone density of mice. After 4 and 6 weeks of treatment, the percentage of muscle weight (ratio of muscle to body weight) and muscle weight / fat weight increased significantly.

[0241] Test Example 3: HFD+CCl4 Model

[0242] Solution preparation: Shake carbon tetrachloride (CCl4) thoroughly, take CCl4 and olive oil (volume ratio = 1:19) in a glass bottle to prepare a 5% CCl4 solution.

[0243] C57BL / 6J mice were induced with 60% HFD for 10 weeks, and the blank control group mice of the same age were fed with ordinary feed for 10 weeks and divided into normal group, model group, and mulberry extract (SZ-A) group. The mulberry extract group was given a dose of 200 mg / kg / d (calculated as alkaloids). The mulberry extract is the mulberry extract prepared in Preparation Example 8. The mice in the model group and the mulberry extract group were intraperitoneally injected with 5% CCl4 solution at 2.5 mL / kg body weight twice a week, and the normal group was injected with the same volume of olive oil (for example: 40 g mice were injected with 100 μL) for 4 weeks. Among them, the normal group was gavaged with normal saline, the drug-treated group was given drugs according to Table 2 below, and the model group was given normal saline. The specific animal grouping and drug treatment are shown in Table 2 below. After the experiment, the femur and tibia of the mice were taken, and the bone density was detected using a bone densitometer iNSiGHTVET DXA / OT20-2F7113-01. The bone density test results of each group are shown in Figure 5 The femur was stained with H&E, and the results are shown in Figure 6 .

[0244] Table 2 Experimental groups

[0245]

[0246] Figure 5 The results showed that the bone density of mice in the model group was significantly reduced, while the bone density of mice in the SZ-A200 administration group tended to increase.

[0247] Figure 6 The results showed that the trabecular edges of the femoral bone tissue in the model group were rough, the trabecular gaps were enlarged, the trabeculae were unevenly arranged and had breakpoints, and the number of fat vacuoles in the bone marrow cavity increased. The number of trabeculae increased and the fat vacuoles decreased in the SZ-A200 administration group.

[0248] Experimental Example 4 Pharmacodynamics of mulberry extract on LZ+HFD-induced endocrine disorder model

[0249] Mulberry extract of Preparation Example 1

[0250] 1. Drug preparation

[0251] The preparation concentration of letrozole (abbreviated as LZ) is: 0.25 mg / mL, and the dosage volume is: 0.4 mL / 100 g.

[0252] 1% CMC: Weigh 1g CMC, add it into 100mL pure water, place it on a magnetic stirrer, set the temperature at 60℃, and the speed at 500r / min, and stir until the CMC is completely dissolved and becomes transparent.

[0253] 2. Experimental Grouping

[0254] Table 3

[0255]

[0256] 3. Experimental methods

[0257] Thirty 3-week-old female SD rats in good health and shiny fur were selected and adaptively fed for 3 days, and randomly divided into blank, model and treatment groups. The model group and the treatment group were given a high-fat diet HFD (research diets D12492)) for 3 weeks, and then gavaged with LZ (1 mg / kg / d) for 6 weeks, and HFD was given at the same time to construct a rat endocrine disorder model. The treatment group was administered according to the dose and method in the table. During the administration period, the blank group and the model group were given an equal amount of normal saline. After 21 days of treatment, the body weight was weighed and the right calf gastrocnemius muscle was weighed.

[0258] Results Figure 7 The results showed that compared with the model group, the muscle weight percentage (ratio of muscle to body weight) of the SZ-A groups at each dose was significantly increased.

[0259] Experimental Example 5 STZ combined with high fat and high cholesterol induced Apoe diabetic sexual dysfunction model in mice

[0260] Thirty 8-week-old Apoe mice were adaptively fed for one week, and then fed with a high-fat and high-cholesterol diet (HFHC) for 5 weeks. STZ (streptozotocin) (100 mg / kg / d) was intraperitoneally injected once on the 6th week to construct a mouse model of lipid metabolism disorder. At the same time, 10 8-week-old C57 mice were adaptively fed for one week, fed with a normal diet for 5 weeks, and injected intraperitoneally with an equal amount of solvent once on the 6th week as the normal control group.

[0261] 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 in Preparation Example 8, and the gastrocnemius muscles of the mice were taken and fixed in 4% paraformaldehyde solution, embedded in paraffin, and sliced ​​for H&E staining. A typical field of view was selected for each sample, and photos were taken under a 200x microscope to measure the average circumference, average diameter, and average area of ​​the cross-section of the muscle fibers in the picture.

[0262] Table 4 Animal grouping and drug administration

[0263]

[0264] The results are as follows Figure 8As shown in A, HE staining results showed that the gastrocnemius muscle in the normal group had a clear structure, and no abnormalities were found in the muscle bundles and muscle fiber gaps. In the model group, muscle fiber edema and atrophy coexisted, and the cross-sections of muscle fibers in the same muscle bundle were significantly different and varied in size. Compared with the model group, the muscle fiber structure was clear after SZ-A administration, and edema and muscle fiber atrophy were alleviated. Figure 8 BD results showed that compared with the model group, the average cross-sectional area, average diameter and average circumference of the gastrocnemius muscle fibers in the drug-treated group were significantly increased.

[0265] Experimental Example 6 Mouse Experiment

[0266] Animal grouping and treatment

[0267] 15-month-old C57 mice were randomly divided into natural aging group, SZ-A administration group and positive drug group, with 15 mice in each group, and the drug was administered once a day by gavage. 3-month-old C57 mice were used as control.

[0268] G1 (normal group): 3-month-old C57 mice, with normal diet and drinking water, and gavage with distilled water as control.

[0269] G2 (natural aging group): 15-month-old C57 mice, with normal diet and drinking water, and gavage with distilled water as control.

[0270] G3: 15-month-old C57 mice were fed with normal diet and drinking water, and were given 100 mg / kg / day (calculated as alkaloids) of the mulberry extract in Preparation Example 1 for 5 months.

[0271] G4: 15-month-old C57 mice were fed a normal diet and drinking water, and were given 200 mg / kg / day (calculated as alkaloids) of the mulberry extract in Preparation Example 1 for 5 months.

[0272] G5: 15-month-old C57 mice were fed with normal diet and drinking water, and were given 100 mg / kg / day (calculated as alkaloids) of the mulberry extract in Preparation Example 7 for 5 months.

[0273] G6: 15-month-old C57 mice were fed with normal diet and drinking water, and were given 200 mg / kg / day (calculated as alkaloids) of the mulberry extract in Preparation Example 7 for 5 months.

[0274] G7 (NMN group, i.e., positive drug group): 15-month-old C57 mice were fed a normal diet and drinking water and given 300 mg / kg / day NMN for 5 months.

[0275] At the end of the experiment, the experimental mice were anesthetized intramuscularly with Shutai, and placed prone in a FAXITRON ULTRAFOCUS 100 bone density analyzer to detect and analyze the bone density of the mice.

[0276] The results are as follows Fig. 9As shown, the results showed that compared with the normal group, the bone density of mice in the aging group was significantly decreased; compared with the aging group, the bone density of the mulberry extract group was significantly increased, and the effect was better than that of the NMN group.

[0277] Experimental Example 7: Feeding db / db diabetic mouse model with normal diet

[0278] The mulberry extract used in the experiment is the mulberry extract in Preparation Example 1.

[0279] 60 5-week-old db / db male mice were purchased, and 20 db / m male mice were used as the normal control group. The mice were kept at room temperature of 22°C, humidity of 60%, and light-dark cycle of 12 hours. After 1 week of adaptive feeding with normal maintenance feed, 10 μL of blood was collected from the tail tip to measure random blood glucose (PBG), and the mice were weighed. According to the levels of random blood glucose and body weight, 60 male db / db mice were divided into a model group (db / db), a low-dose group of mulberry extract (SZ-A) SZ-A100 (100 mg / kg / d, calculated as alkaloids), and a medium-dose group of mulberry extract (SZ-A) SZ-A200 (200 mg / kg / d, calculated as alkaloids), with 20 mice in each group; 20 5-week-old db / m mice were used as the normal group. All mice were fed with basal feed. After 2 weeks of adaptive feeding, each experimental group was treated with the corresponding dose of the test drug for 8 consecutive weeks. The basal feed was continued during the treatment period, and the body weight and blood sugar of the animals were monitored. The model group and the normal group were given normal saline. After 8 weeks of administration, the grip strength meter was used to evaluate the effect of SZ-A on the muscle strength of diabetic mice. The grip strength meter (Yuyan Instrument) consists of a gripping rod and a high-precision force measurement system. Place the forelimbs of the experimental mouse on the horizontal bar of the gripping rod, grab the mouse tail and gently pull it back. After the mouse grasps the gripping rod, pull it back evenly to cause the mouse to loosen its claws. The instrument automatically records the maximum grip of the animal, and the measurement is repeated many times to ensure the result, which represents its muscle strength.

[0280] The results of mouse grasping strength are as follows Fig.10 The results showed that the gripping strength of diabetic mice in the model group was significantly reduced, and after 8 weeks of treatment, the gripping strength of mice in the SZ-A 100 mg / kg / d and 200 mg / kg / d groups was significantly increased. This indicates that mulberry extract can significantly improve the muscle strength of db / db diabetic mice.

[0281] Test Example 8:

[0282] Inhibitory effect of mulberry extract on muscle cell damage

[0283] Growth medium (high glucose DMEM medium + 10% fetal bovine serum) was added to C2C12 cells and incubated in a constant temperature incubator at 37°C and 5% CO2. The cells were cultured to about 85%, and the cells were collected and grouped into a control group (NC), a model group (D-gal), and an SZ-A group (D-gal + SZ-A). Subsequently, the culture medium of each group was replaced with a differentiation medium (high glucose DM EM medium + 2% horse serum). According to the purpose of the experiment, D-gal and SZ-A (SZ-A obtained in Preparation Examples 2, 6, and 8) were added to the differentiation medium before the end of the experiment. D-gal at a concentration of 100mM was added to the culture medium of the model group, and SZ-A at a concentration of 100ug / ml and 100mM D-gal were added to the culture medium of the SZ-A group for 48h. SA-β-galactosidase staining was used to detect the degree of damage to cells in different groups.

[0284] Group 1: control group;

[0285] Group 2: Model group;

[0286] Group 3: SZ-A group, SZ-A is the mulberry branch extract in Preparation Example 2;

[0287] Group 4: SZ-A group, SZ-A is the mulberry branch extract in Preparation Example 6;

[0288] Group 5: SZ-A group, SZ-A is the mulberry branch extract in Preparation Example 8;

[0289] Fig.11 The results showed that SZ-A could inhibit the damage of C2C12 muscle cells induced by D-gal.

[0290] Experimental Example 9: Study on the distribution of total alkaloids from Morus alba in bone marrow tissue

[0291] 1. Medicines:

[0292] Miglitol reference substance (Lot: HRSLW-DM, content 100.1%) TCI (Shanghai) Chemical Industry Development Co., Ltd.

[0293] DNJ reference substance (Lot: 20171024, content: 99.29%) Beijing Wuhe Bio Pharmaceutical Co., Ltd.

[0294] FA reference (Lot: 16411, content: 100%) MedChemExpress

[0295] DAB reference (Lot: 0000161756, content: 100%) Sigma

[0296] 2. Solution preparation

[0297] Preparation of dosing solution: accurately weigh 1.65 g of mulberry extract, add appropriate amount of ultrapure water to dissolve, and shake well to prepare a mulberry extract oral administration solution with a concentration of 4 mg / ml (calculated as alkaloids).

[0298] Preparation of the assay solution: Preparation of 0.1% ammonia water as the mobile phase: Accurately pipette 4 ml of ammonia water (25%), add 1000 ml of water, shake well, filter, and degas by ultrasonication for 20 min.

[0299] Preparation of stock solution: Accurately weigh 17.0 mg of DNJ reference substance, 8.5 mg of FA reference substance, 8.5 mg of DAB reference substance, and 10.0 mg of miglitol reference substance, respectively place them in 10 ml volumetric flasks, dissolve them with Wahaha water, make up to volume, and shake well to obtain stock solutions with DNJ concentration of 1.688 mg·ml-1, FA concentration of 0.850 mg·ml-1, DAB concentration of 0.850 mg·ml-1, and miglitol concentration of 1.0 mg·ml-1.

[0300] Preparation of mixed reference substance working solution for rat bone marrow tissue assay:

[0301] Accurately measure appropriate amounts of DNJ, FA, and DAB stock solutions respectively, mix them in a 5 ml volumetric flask, and dilute to volume with methanol-water (4:1, v / v) to obtain a DNJ concentration of 25000 ng ml -1 , FA and DAB concentrations were 12500 ng·ml -1 The standard working solution was diluted in series to obtain DNJ concentrations of 125, 250, 625, 1250, 2500, 6250, and 12500 ng / ml, and FA and DAB concentrations of 62.5, 125, 312.5, 625, 1250, 3125, and 6250 ng·ml -1 Mix the standard curve working solution in different concentration series.

[0302] Preparation of rat bone marrow tissue quality control solution:

[0303] Accurately measure appropriate amounts of DNJ, FA, and DAB stock solutions respectively, mix them in a 5 ml volumetric flask, and dilute to volume with methanol-water (4:1, v / v) to obtain a DNJ concentration of 25000 ng ml -1 , FA and DAB concentrations were 12500 ng·ml -1 The standard working solution was diluted in series to obtain DNJ concentrations of 50, 100, 1250, and 10000 ng ml -1 , FA and DAB concentrations were 125, 625, and 5000 ng·ml -1 Low, medium and high concentration quality control solutions.

[0304] 3. Animal medication and material collection

[0305] 24 6-8 week old SD rats (180-200g), half male and half female. SD rats were fasted for 12h before administration and had free access to water. Rats were orally gavaged with 40mg / kg (calculated as alkaloids) of mulberry extract and then killed at 5min, 30min, 4h, and 12h. Six rats (half male and half female) were killed at each time point. The bone marrow (femur) of the rats was taken and stored in a -80℃ refrigerator.

[0306] Bone marrow tissue sample processing: The rat bone marrow tissue samples were placed in a freezing grinder for homogenization. The freezing grinder was turned on 30 minutes in advance and refrigerated to 4°C. The homogenization parameters were: frequency 60 Hz, running time 60 seconds, 2 times, and interruption time 5 seconds. The tissue homogenate was prepared and set aside.

[0307] Take 50 μl of rat bone marrow homogenate, add 10 μl of methanol-water (4:1, v:v) solution and 200 μl of 100 ng ml -1 The protein precipitation solvent of the internal standard was vortexed for 2 minutes, placed at 4°C for 10 minutes, centrifuged at 14000rpm×10min, 100μl of the supernatant was taken, diluted to 200μl with 75% acetonitrile (0.1% formic acid), vortexed for 1 minute, centrifuged at 14000rpm for 5 minutes, and the supernatant was taken for sampling and determination.

[0308] 4. LC-MS / MS chromatography and mass spectrometry conditions

[0309] 4.1 Chromatographic conditions

[0310] The chromatographic column was an XBridgeTM Amide column (3.5 μm, 4.6 mm×150 mm, Waters, USA); mobile phase A was water (0.1% ammonia water), phase B was acetonitrile, gradient elution: 0.0-4.0 min 65% mobile phase B, 4.0 min-7.0 min 43% mobile phase B, 7.0 min-13.0 min 65% mobile phase B; flow rate was 0.5 ml / min; column temperature was 35°C; injector temperature was 15°C; injection volume was 5 μl.

[0311] 4.2 Mass spectrometry conditions

[0312] Electrospray ion source (ESI), Curtain Gas (CUR): 35.0, Collision Gas (CAD): 9, IonSprayVoltage (IS): 5500, Temperature (TEM): 500, IonSource Gas1 (GS1): 55.0, IonSource Gas2 (GS2) 50.0, Entrance Potential (EP): 10.0; Collision Cell Exit Potential (CXP): 6.0, positive ion scanning mode was used, and the scanning method was multiple reaction monitoring (MRM). The reaction ions of DNJ, FA, DAB and miglitol (internal standard, IS) used for quantitative analysis were: m / z 164.3→69.0 (DP 70ev, CE 27ev), m / z 148.2→112.1 (DP 56ev, CE 18ev), m / z 134.3→68.1 (DP 55ev, CE 25ev), m / z208.3→146.1 (DP 70ev, CE 27ev).

[0313] 5. Results statistics and analysis

[0314] Calculate the drug concentration in bone marrow tissue, as shown in Table 4, and draw a bone marrow drug tissue distribution column Fig.12 .

[0315] Table 4 Drug concentration in bone marrow tissue of rats given total alkaloids of Morus alba 40 mg / kg by oral administration (ng / g, n=6)

[0316]

[0317] 6. Results and Discussion

[0318] The results showed that after rats were orally gavaged with mulberry extract, alkaloids could be detected in the bone marrow tissue within 5 minutes. The distribution trends of DNJ, FA and DAB in the bone marrow tissue were consistent, reaching the maximum drug concentration in 0.5 hours and were basically eliminated from the bone marrow tissue after 12 hours of administration, indicating that there was no risk of accumulation of SZ-A in the bone marrow tissue and that it was relatively safe.

Claims

1. Use of mulberry extract in the preparation of a medicament for treating and / or preventing diseases related to abnormal bone metabolism, characterized in that: Taking the weight percentage of the sum of the components of the mulberry extract as 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 comprising a polysaccharide content of not more than 70% by weight (optionally comprising a polysaccharide content of 0.2-70% by weight, further optionally comprising a polysaccharide content of 0.2-50% by weight, further optionally comprising a polysaccharide content of 0.2-35% by weight, further optionally comprising a polysaccharide content of 0.2-25% by weight, further optionally comprising a polysaccharide content of 0.2-23% by weight, further optionally comprising a polysaccharide content of 20-25% by weight), and / or contains flavonoids having a weight content of not more than 10% (optionally containing flavonoids having a weight content of 0.05-5%, further optionally containing flavonoids having a weight content of 0-2%, further optionally containing flavonoids having a weight content of 0.05-2%, further optionally containing flavonoids having a weight content of 0.5-1.5%, further optionally containing flavonoids having a weight content of 0-1%, further optionally containing flavonoids having a weight content of 0.05-1%), and / or contains no more than 50% amino acids by weight (optionally contains 0-30% amino acids by weight, further optionally contains 0-25% amino acids by weight, further optionally contains 0-20% amino acids by weight, further optionally contains 0-5% amino acids by weight, further optionally contains 3-25% amino acids by weight or further optionally contains 5-20% amino acids by weight), 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 disease related to abnormal bone metabolism is any one of osteoporosis, sarcopenia, sarcopenia-osteoporosis, muscular dystrophy, myasthenia, osteomalacia, rickets, primary hyperparathyroidism bone disease, fluorosis, osteoarthritis and bone displacement; it can be selected from osteoporosis, sarcopenia or sarcopenia-osteoporosis.

3. The use according to claim 1 or 2, characterized in that: The use of the mulberry extract in the preparation of a drug for treating and / or preventing diseases related to abnormal bone metabolism includes: the use of the mulberry extract in any one of the following (b1)-(b3): (b1) Use in the preparation of a drug for increasing bone density; (b2) Use in the preparation of a drug for reducing bone marrow fat cavitation; (b3) Use in the preparation of drugs for improving bone microstructure; The improvement of bone microstructure includes at least one of the following: (i) increasing the number of trabeculae; (ii) reducing the separation of trabeculae; (iii) reducing the trabecular structure model index; (iv) increasing the bone volume fraction; And / or for use in any one of the following (1)-(7): 1) Use in the preparation of drugs for increasing muscle mass; 2) Use in the preparation of drugs for increasing muscle weight percentage; 3) Use in the preparation of drugs for increasing muscle fat ratio; 4) Use in the preparation of a drug for inhibiting muscle atrophy and / or edema; optionally, the inhibiting muscle atrophy and / or edema is specifically to increase the cross-sectional area, muscle fiber diameter and / or muscle fiber circumference of the muscle fiber; 5) Use in the preparation of drugs for relieving intermuscular peripheral neuropathy and / or vascular lipid deposition; 6) Use in the preparation of drugs for inhibiting muscle cell damage; 7) Use in the preparation of drugs for increasing muscle strength.

4. The use according to any one of claims 1 to 3, characterized in that: Taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is: Optionally, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is: Optionally, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is: Optionally, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is: Optionally, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is: Optionally, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is: Optionally, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is: Further optionally, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is: Further optionally, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is: Further optionally, taking the sum of the weight percentage of the components of the mulberry extract as 100%, the weight content of the components in the mulberry extract is:

5. The use according to any one of claims 1 to 4, characterized in that The alkaloids include 1-deoxynojirimycin (DNJ), N-methly-1-deoxynojirimycin, fagomine (FAG), 3-epi-fagomine, 1,4-dideoxy-1,4-imino-D-arabinitol (DAB), calystegin B2, calystegin C1, 2-oxo-(α-D-galactoside) and 1,4-dideoxy-1,4-imino-D-arabinitol (DAB). 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; Optionally, the weight percentage of DNJ is not less than 50% (optionally 60-99%) of the total alkaloids.

6. The use according to any one of claims 1 to 5, characterized in that: The preparation method 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 the mulberry extract.

7. The use according to claim 6, characterized in that: The method further comprises the steps of: 3) performing alcohol precipitation on the resin effluent of step 2) and collecting the supernatant; 4) concentrating and / or drying the supernatant; Optionally, the method further comprises the following step: concentrating and / or drying the resin effluent from step 2).

8. The use according to any one of claims 1 to 7, characterized in that: The mulberry extract acts on humans or mammals.

9. The use according to any one of claims 1 to 8, characterized in that: The drug is in the form of an oral administration 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.

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