Application of quercetin-3-caffeoyl robin glucoside in preparation of product for preventing and treating osteoporosis

Abstract

The invention provides an application of a compound quercetin-3-caffeoyl robinoside (Q3C) and a salt or a carrier of the compound quercetin-3-caffeoyl robinoside (Q3C) in preparation of an anti-osteoporosis product, and belongs to the technical field of medical application, and the compound quercetin-3-caffeoyl robinoside (Q3C) and the salt or the carrier of the compound quercetin-3-caffeoyl robinoside (Q3C) can be used for preventing osteoporosis. An in-vitro osteoblast model and an in-vivo ovariectomized osteoporosis mouse model prove that the Q3C can promote proliferation and differentiation of bone cells and increase synthesis and mineralization of bone matrixes, so that bone formation of osteoporosis mice is promoted, the bone density is improved, the fracture risk is reduced, and the Q3C can be used as an active component for preparing anti-osteoporosis products.

Description

Technical Field

[0001] The present invention relates to the technical field of medical applications, and in particular to the application of quercetin-3-caffeoylrobin glycoside and a pharmaceutically acceptable salt or carrier thereof in the preparation of a product for preventing and / or treating osteoporosis. Background Art

[0002] Osteoporosis is a chronic metabolic bone disease characterized by a higher degree of bone resorption than bone formation, leading to decreased bone mass and damaged bone microstructure. Osteoporosis seriously affects the quality of life of the elderly and imposes a great economic burden on society.

[0003] Bone remodeling is a complex process involving the interaction of multiple cells. The main process is osteoclast-mediated bone resorption to absorb old bone, while osteoblast-mediated bone formation produces new bone. Currently, various drugs for the treatment of osteoporosis have been shown to reduce the risk of fractures, including bisphosphonates that inhibit osteoclast bone resorption, selective estrogen receptor modulators, monoclonal antibodies RANKL and parathyroid hormone, but these drugs have certain side effects, such as causing abdominal pain, nausea, vomiting, joint pain and other symptoms, and have great limitations in practical application.

[0004] Nowadays, natural products have been used as important, safe and convenient resources. Many natural medicines extracted from nature have been used to treat osteoporosis. For example, lycopene extracted from ripe red fruits (tomatoes, carrots, etc.) can effectively reduce bone loss and bone resorption markers in postmenopausal women. Genistein, which was previously reported in studies, is present in soybeans and their products. It is a selective estrogen receptor modulator that can bind to estrogen receptor β to treat osteoporosis caused by postmenopausal women. It not only effectively increases bone density, but is also safe and non-toxic. Therefore, it is of great significance to develop new potential natural active substances as effective drugs for the treatment of osteoporosis.

[0005] Quercetin is a naturally occurring flavonoid compound, which is mostly present in the flowers, leaves and fruits of many plants in the form of glycosides, and has the advantages of high safety and few adverse reactions. So far, there are many reports showing that quercetin has multiple pharmacological activities such as anti-inflammatory, anti-oxidant, anti-viral, immunomodulatory, and cardiovascular protection. Studies have also shown that quercetin can maintain the balance between osteoblastogenesis and osteoclastogenesis, plays an important role in the treatment of osteoporosis, and its partial derivatives, such as patent CN115969867A discloses that quercetin is also confirmed to have an anti-osteoporotic effect. But its derivative quercetin-3-caffeylrobinobioside (Quercetin 3-Caffeylrobinobioside, Q3C) can be used for the related research of preventing and treating osteoporosis, which has not been reported yet. Summary of the invention

[0006] In order to solve the above problems, this application proposes the following technical solutions:

[0007] On the one hand, the present application provides a derivative of quercetin, quercetin-3-caffeylrobinobioside (Quercetin3-Caffeylrobinobioside, Q3C), or a pharmaceutically acceptable salt thereof, or a derivative thereof, or a pharmaceutically acceptable carrier containing Q3C as an active ingredient in the preparation of a product for preventing and / or treating osteoporosis.

[0008] The compound name of the quercetin-3-caffeoyl rhamnoside described in the present application is: quercetin-3-O-[4-O-trans-caffeoyl-ALPHA-L-rhamnosyl-(1→6)-BETA-D-galactoside, with a molecular formula of C 36 H 36 O 19 , with a molecular weight of 772.66, and having a structural formula as shown in formula (I):

[0009]

[0010] In one embodiment, the active ingredient in the product is selected from at least one of the following:

[0011] A), quercetin-3-caffeoylrobin glycoside having a structural formula as shown in formula (I);

[0012] B) a pharmaceutically acceptable salt of quercetin-3-caffeoylrobin glycoside as described in A);

[0013] C), a pharmaceutically acceptable carrier loaded with quercetin-3-caffeoylrobin glycoside as described in A).

[0014] In one embodiment, the pharmaceutically acceptable salt is selected from any one or more of its hydrochloride, sulfate, acetate, methanesulfonate, succinate, fumarate, citrate, malate or organic amine salt.

[0015] In one embodiment, the pharmaceutically acceptable carrier includes an inorganic carrier or a biological carrier.

[0016] In one embodiment, the vector includes, but is not limited to, a plasmid, a liposome, an extracellular vesicle or a viral vector, or a bone-targeted nanocarrier.

[0017] Optionally, the bone-targeted nanocarriers include, but are not limited to, bone-targeted nanohydrogels, nanoparticles, nanomicrospheres, and nanomicelles.

[0018] In one embodiment, the quercetin-3-caffeoylrobin glycoside is derived from animal and plant extraction, chemical synthesis or microbial fermentation.

[0019] Optionally, the Q3C can be obtained by an extraction method in plants and fruits and vegetables, and the plants and fruits and vegetables can be one or more of Sophora japonica seeds, onions, apples, grapes, berries, broccoli, citrus fruits, cherries, tea and capers.

[0020] Optionally, the Q3C can be obtained through chemical synthesis methods such as esterification reaction, glycosidation reaction and purification process.

[0021] In one embodiment, the product is selected from medicine, food, functional food or special medical food.

[0022] In one embodiment, the content of active ingredients in the product is 0.01%-99.99%, preferably 0.05%-95%; more preferably 0.05%-65%; the % is the mass volume percentage (w / v) or mass percentage (w / w).

[0023] In one embodiment, the product is for use in promoting bone formation.

[0024] Preferably, the promotion of bone formation is the promotion of proliferation and differentiation of osteoblasts.

[0025] In one embodiment, the osteoblasts comprise MC3T3-E1 cells.

[0026] In one embodiment, the Q3C is used to increase the level of alkaline phosphatase (ALP) in osteoblasts.

[0027] In one embodiment, the concentration of Q3C used to promote osteoblast differentiation is 0.01-10 μM, preferably 0.02-10 μM, more preferably 1-10 μM, for example, 1 μM, 1.5 μM, 2 μM, 2.5 μM, 3 μM, 3.5 μM, 4 μM, 4.5 μM, 5 μM, 5.5 μM, 6 μM, 6.5 μM, 7 μM, 7.5 μM, 8 μM, 8.5 μM, 9 μM, 9.5 μM, and 10 μM.

[0028] In one embodiment, the osteoporosis includes postmenopausal osteoporosis and / or senile osteoporosis.

[0029] In one embodiment, the product is used to improve the decrease in bone mass, bone calcium content and / or bone density caused by osteoporosis.

[0030] In one embodiment, the quercetin-3-caffeoylrobin glycoside in the product can also be prepared into a compound preparation with ingredients having the same activity or other activities, and the other active ingredients are selected from anti-inflammatory, antioxidant, antibacterial, antiviral, antitumor, immunomodulatory and / or cardiovascular protective active ingredients.

[0031] Optionally, the active ingredients for treating osteoporosis include, but are not limited to, bisphosphonates, raloxifene, calcitonin, teripine, denosumab, estrogen, romosozumab, vitamin D, calcium, hyaluronic acid, chondroitin sulfate, and glucosamine.

[0032] Optionally, the other active ingredients include but are not limited to: vitamins, quercetin, resveratrol, anthocyanidins, curcumin, and green tea polyphenols.

[0033] In one embodiment, the dosage form of the product includes injection, suspension, granules, capsules, powders, tablets, pills, suppositories, aerosols or drops.

[0034] In one embodiment, the administration route of the product includes intravenous injection, intraperitoneal injection, intramuscular injection, subcutaneous injection, oral administration, sublingual administration, nasal administration or aerosol administration.

[0035] On the other hand, the present application also provides the use of quercetin-3-caffeoylrobin glycoside and its pharmaceutically acceptable salt or carrier as an active ingredient in the preparation of an agent for promoting the proliferation and differentiation of osteoblasts, increasing animal bone dry weight, bone calcium content and / or bone density.

[0036] On the other hand, the present application also provides a drug for preventing and treating osteoporosis, comprising an active ingredient containing quercetin-3-caffeoylrobin glycoside and at least one pharmaceutically acceptable excipient.

[0037] In one embodiment, the pharmaceutically acceptable excipients include, but are not limited to, diluents, excipients, fillers, binders, wetting agents, disintegrants, sustained-release agents, absorption enhancers, surfactants, adsorption carriers, and lubricants.

[0038] In one embodiment, the content of Q3C in the drug is 0.01%-99.99%, wherein the % is the mass volume percentage (w / v) or the mass percentage (w / w).

[0039] In one embodiment, the drug may be a tablet or a suspension, wherein the content of Q3C may be 0.01%-99.99%, preferably 0.05%-95%, more preferably 0.05%-65%.

[0040] In other embodiments, the drug can be in various dosage forms such as oral dosage forms, and the content of Q3C in other dosage forms can be added according to the conventional active ingredient addition amount of the dosage form.

[0041] Optionally, the working concentration of Q3C in the drug is 10 to 250 mg / kg.

[0042] Optionally, the working concentration of Q3C in the drug is 25-100 mg / kg.

[0043] Optionally, the suitable daily dosage range of Q3C is 25-100 mg / kg body weight. The above dosage can be administered in one dosage unit or divided into several dosage units, depending on the clinical experience of the doctor and the dosage regimen including the use of other treatment methods. In the present invention, when administering to animals, the daily dosage of Q3C and / or its pharmaceutically acceptable salt and / or its carrier is 25-100 mg / kg mouse, and the administration period is 6-18 weeks.

[0044] This application has at least the following technical advantages or beneficial effects:

[0045] This application studies the therapeutic effect of Q3C on osteoporosis through an ovariectomy (OVX)-induced mouse osteoporosis model and an in vitro mouse skull isolated osteoblast precursor cell (MC3T3-E1) model. The experimental results show that Q3C can effectively promote the proliferation of osteoblasts MC3T3-E1 in vitro, increase the ALP content of MC3T3-E1 cells, increase the synthesis and mineralization of bone matrix, and Q3C can also effectively improve the bone dry weight, bone calcium content and bone density caused by osteoporosis in animals, reduce the risk of fractures, and can be used as an active ingredient to produce new, more effective and safe products for the treatment of osteoporosis, while laying the foundation for further research on new synergistic pharmaceutical compositions. BRIEF DESCRIPTION OF THE DRAWINGS

[0046] The accompanying drawings, which are incorporated in and constitute a part of this specification, illustrate embodiments consistent with the invention and, together with the description, serve to explain the principles of the invention.

[0047] In order to more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the following briefly introduces the drawings required for use in the embodiments or the prior art descriptions. For ordinary technicians in this field, other drawings can be obtained based on these drawings without creative work. In the drawings:

[0048] Figure 1 This is the result of the effect of different concentrations of Q3C on the proliferation of mouse embryonic osteoblast precursor cells (MC3T3-E1) in vitro in Test Example 3;

[0049] Figure 2 The figure is the result of the effect of different concentrations of Q3C on the alkaline phosphatase (ALP) level of MC3T3-E1 cells in test example 4;

[0050] Figure 3 This is a graph showing the effects of different concentrations of Q3R on the alkaline phosphatase (ALP) level of MC3T3-E1 cells in Test Example 4. DETAILED DESCRIPTION

[0051] In order to more clearly explain the overall concept of the application, the following is described in detail by way of embodiment. In the following description, a large amount of specific details are provided so as to provide a more thorough understanding of the application. However, it is obvious to those skilled in the art that the application can be implemented without one or more of these details. In other examples, in order to avoid confusion with the application, some technical features well known in the art are not described.

[0052] Unless otherwise specified, in the following embodiments, reagents or instruments used without indicating the manufacturer are all conventional products that can be purchased from the market. If no specific conditions are indicated in the examples, conventional conditions or conditions recommended by the manufacturer are followed.

[0053] In the following examples, some reagents and instrument information are shown in Table 1:

[0054] Table 1

[0055]

[0056]

[0057] Example 1

[0058] This embodiment provides a Q3C tablet (100 tablets), and the preparation steps are as follows: weigh 30g of powdered sugar, 45g of microcrystalline cellulose and 8g of sodium carboxymethyl starch, mix them evenly, weigh 10mg of Q3C, mix them evenly with 12g of 10% starch slurry and add them as a binder to prepare a soft material, pass through a 14-mesh nylon sieve to prepare wet granules, dry at 45°C, add talcum powder, mix evenly, pass through a 12-mesh nylon sieve, and use a single-punch tablet press to tablet the granules after passing the content test to obtain Q3C tablets with a content of 0.01%.

[0059] Example 2

[0060] This embodiment provides a Q3C tablet (100 tablets), and the preparation method thereof is as shown in Example 1, except that the mass percentage of Q3C in the tablet prepared in this embodiment is 0.05%.

[0061] Example 3

[0062] This embodiment provides a Q3C tablet (100 tablets), and the preparation method thereof is as shown in Example 1, except that the mass percentage of Q3C in the tablet prepared in this embodiment is 0.1%.

[0063] Example 4

[0064] This embodiment provides a Q3C tablet (100 tablets), and the preparation method thereof is as shown in Example 1, except that the mass percentage of Q3C in the tablet prepared in this embodiment is 0.5%.

[0065] Example 5

[0066] This embodiment provides a Q3C tablet (100 tablets), and the preparation method thereof is as shown in Example 1, except that the mass percentage of Q3C in the tablet prepared in this embodiment is 2.5%.

[0067] Example 6

[0068] This embodiment provides a Q3C tablet (100 tablets), and the preparation method thereof is as shown in Example 1, except that the mass percentage of Q3C in the tablet prepared in this embodiment is 5%.

[0069] Example 7

[0070] This embodiment provides a Q3C tablet (100 tablets), and the preparation method thereof is as shown in Example 1, except that the mass percentage of Q3C in the tablet prepared in this embodiment is 10%.

[0071] Example 8

[0072] This embodiment provides a Q3C suspension, which is prepared by conventional procedures, and the specific process is as follows:

[0073] Weigh 150g of Q3C powder and mix it with an appropriate amount of polyvinyl pyrrolidone, and use a small amount of purified water to moisten it. Then mix hydroxyethyl cellulose and a small amount of water and stir until it is completely dissolved and forms a viscous liquid. Finally, add the two into the blender, adjust the pH to 7.0, slowly add purified water to 500mL, and stir magnetically for 10 minutes to ensure that Q3C is evenly dispersed. Package into 100 pieces, press the stopper, unpack, cover, and label and package after inspection to obtain a Q3C suspension with a mass percentage of 30%.

[0074] Example 9

[0075] This embodiment provides a Q3C suspension, and the preparation method thereof is as shown in Example 8, except that the mass percentage of Q3C in the suspension prepared in this embodiment is 65%.

[0076] Example 10

[0077] This embodiment provides a Q3C suspension, and the preparation method thereof is as shown in Example 8, except that the mass percentage of Q3C in the suspension prepared in this embodiment is 95%.

[0078] Embodiment 11

[0079] This embodiment provides a Q3C suspension, and the preparation method thereof is as shown in Example 8, except that the mass percentage of Q3C in the suspension prepared in this embodiment is 99.99%.

[0080] Quercetin QUE group

[0081] A tablet containing quercetin was prepared by the method shown in Example 1, and the mass percentage was 10%. The molecular formula of QUE is: 15 H 10 O7, with a molecular weight of 302.24, has a structural formula as shown in formula (II):

[0082]

[0083] Quercetin derivatives Q3R group

[0084] Tablets containing another quercetin derivative, quercetin 3,7-di-O-rhamnoside (Q3R), were prepared by the method shown in Example 1, and the mass percentage was 10%. The molecular formula of Q3R is: C 27 H 30 O 15 , with a molecular weight of 594.52, and having a structural formula as shown in formula (III):

[0085]

[0086] Test Example 1

[0087] This test example verifies that the tablets in the above embodiment can prevent bone loss in the postmenopausal osteoporosis mouse model. The specific experimental process is as follows:

[0088] A number of 8-week-old female SPF-grade C57BL / 6J mice weighing 18-22 g were selected and randomly divided into: a sham operation group (Sham group), an ovariectomy group (ovariectomy, OVX group), Example 1-7 groups, a QUE group, and a Q3R group, with 10 mice in each group. Except for the Sham group, the mice in other groups underwent bilateral ovariectomy to construct an osteoporosis mouse model. The specific operation method is as follows:

[0089] The mice were anesthetized by intraperitoneal injection of 30 mg / kg BW of pentobarbital solution. After sufficient anesthesia, the lower abdomen of the mice was prepared, and a longitudinal incision of about 1.5 cm was made along the midline of the abdomen in the middle of the lower abdomen. The fat layer was pushed aside to find the uterus, and the ovary wrapped in fat mass was visible at its end. Then the fallopian tube under the ovary (including fat) was ligated with silk thread, and the ovary was removed. The other ovary was removed in the same way. After the abdominal muscles and skin were sutured in layers, they were disinfected again. The sham group mice were exposed using the same surgical procedure but the bilateral ovaries were not removed. Only part of the fat was removed for sham surgery. After surgery, each mouse was injected with penicillin solution for 3 consecutive days to prevent wound infection. All surgical operations were performed under sterile conditions.

[0090] After 3 days of recovery from bilateral ovariectomy, drug intervention began. The mice in the Sham group and OVX group were gavaged with normal saline, and the mice in the remaining groups were gavaged with corresponding drugs (6 tablets prepared in Examples 1 to 7 and the QUE group and Q3R group were taken, ground into powder with a grinder, and 10 mL of pure water was added to mix to obtain the corresponding gavage drug for each example). The gavage volume was 100 μL for three months. After the administration, the mice were killed, and the dry weight of the femur, bone density, and bone calcium content of the mice were measured. The experimental results are shown in Table 2.

[0091] Table 2 Comparison of mouse groups and femoral indexes among mice in each group (n=10, mean±SD)

[0092]

[0093] As shown in Table 2, compared with the Sham group, the femoral dry weight, bone calcium content and bone density of mice in the OVX group were significantly reduced, indicating that the ovariectomized osteoporosis mouse model was successfully established. In addition, compared with the OVX group, the femoral dry weight, bone calcium content and bone density of mice in Example 1 to Example 7 groups after administration of Q3C tablets increased to varying degrees, and were dose-dependent, indicating that oral Q3C can effectively reduce bone loss in postmenopausal osteoporotic mice and has a good preventive or therapeutic effect on postmenopausal osteoporosis.

[0094] At the same time, compared with the OVX group, the femoral dry weight, bone calcium content and bone density of the mice in the QUE group were increased, but compared with the same dose of Example 7 group, the femoral dry weight, bone calcium content and bone density were decreased, indicating that Q3C can better improve bone loss caused by osteoporosis than QUE.

[0095] In addition, compared with the OVX group, the femur dry weight, bone calcium content and bone density of the mice in the Q3R group with the same dose as in Example 7 were slightly increased, indicating that although they are all compounds derived from quercetin, their different quercetin derivatives have great differences in anti-osteoporosis activity.

[0096] Test Example 2

[0097] This test example verifies that the Q3C suspension in the above example can prevent bone loss in the postmenopausal osteoporosis mouse model. The specific process is as follows:

[0098] 60 8-week-old female SPF-grade C57BL / 6J mice weighing 18-22 g were selected and randomly divided into: Sham group, OVX group, and Example 8-11 groups, with 10 mice in each group. Except for the Sham group, the mice in other groups underwent bilateral ovariectomy to construct an osteoporosis mouse model. The specific operation method was the same as that in Test Example 1.

[0099] After the bilateral ovariectomy surgery, the drug intervention was started 3 days after recovery. The mice in the Sham group and OVX group were gavaged with 10 μL of normal saline, and the mice in the remaining groups were gavaged with 10 μL of Q3C suspension prepared in the corresponding example for three months. After the end of the drug administration, the mice were killed, and the dry weight of the femur, bone density, and bone calcium content of the mice were measured. The experimental results are shown in Table 3.

[0100] Table 3 Comparison of mouse groups and femoral indexes among mice in each group (n=10, mean±SD)

[0101]

[0102] From the results in Table 3, it can be seen that compared with the Sham group, the femoral dry weight, bone calcium content and bone density of mice in the OVX group were significantly reduced, indicating that the ovariectomized osteoporosis mouse model was successfully constructed. Compared with the OVX group, the femoral dry weight, bone calcium content and bone density of mice in Examples 8 to 11 increased to varying degrees after being given Q3C suspension, indicating that Q3C suspension can effectively reduce bone loss in postmenopausal osteoporotic mice and has a good preventive or therapeutic effect on postmenopausal osteoporosis. In addition, combining the results of Tables 2 and 3, it can be seen that when the drug concentration exceeds 60 mg / mL, its enhancement effect tends to be gentle.

[0103] Test Example 3

[0104] This test example verifies the effect of Q3C on the proliferation and differentiation of mouse embryonic osteoblast precursor cells (MC3T3-E1) in vitro. The specific process is as follows:

[0105] Effect of Q3C on cell proliferation: MC3T3-E1 cells in the logarithmic growth phase were evenly inoculated in a 96-well plate. When the cell fusion rate reached more than 70%, the old culture medium was discarded and the cells were intervened with complete culture medium containing different concentrations of Q3C (0μM, 0.25μM, 0.5μM, 1μM, 2.5μM, 5μM) for 24 hours. The absorbance value at a wavelength of 405nm was detected by an ELISA instrument to calculate the cell proliferation rate. The complete culture medium consisted of α-MEM, 10% FBS and 1% penicillin-streptomycin. The results are shown in Table 4 and Figure 1 shown.

[0106] Table 4 Effect of Q3C on the proliferation rate of MC3T3-E1 cells (n=5, mean±SD)

[0107] Concentration (μM) Cell proliferation rate (%) 0 101.07376±0.81626 0.25 <![CDATA[117.93953±1.38632 * ]]> 0.5 <![CDATA[117.77973±3.22932 * ]]> 1 <![CDATA[119.2176±3.74183 * ]]> 2.5 <![CDATA[121.16837±9.48175 ** ]]> 5 <![CDATA[120.58817±3.72783 ** ]]>

[0108] From Table 4 and Figure 1 The results in Figure 3 show that Q3C within the selected concentration range can promote the proliferation of MC3T3-E1 cells.

[0109] Test Example 4

[0110] This test example verifies the effect of Q3C on the alkaline phosphatase (ALP) level of MC3T3-E1 cells. The specific process is as follows:

[0111] MC3T3-E1 cells in the logarithmic growth phase were evenly inoculated in a 24-well plate. When the cell fusion rate reached more than 70%, the old culture medium was discarded and the cells were induced to differentiate for 14 days with osteogenic differentiation medium (Induce group, containing 50 μg / mL ascorbic acid and 10 mM β-glycerophosphate) and osteogenic differentiation medium containing different concentrations of Q3C (0.25 μM, 0.5 μM, 1 μM, 2.5 μM, 5 μM). The cells in each group were collected and the ALP level was detected. During the differentiation process, the osteogenic differentiation medium was replaced every 2 to 3 days. The results are shown in Tables 5 and Figure 2 shown.

[0112] Table 5 Effect of Q3C on ALP content in MC3T3-E1 cells (n=5, mean±SD)

[0113] Group ALP content (U / L) Control Group 74.095±3.033 Induce Group <![CDATA[105.109±3.318 ### <!-- 8 -->]]> 0.25μM 99.688±8.492 0.5μM 101.651±2.199 1μM <![CDATA[115.276±5.914 * ]]> 2.5μM <![CDATA[117.047±2.207 ** ]]> 5μM <![CDATA[125.979±6.494 *** ]]>

[0114] From Table 5 and Figure 2The results show that compared with the Control group, the ALP content in the Induce group increased significantly, indicating that MC3T3-E1 cells were successfully induced. Compared with the Induce group, within the concentration range selected in the experiment, when the Q3C concentration was lower than (equal to) 0.5 μM, the ALP content in osteoblasts decreased slightly, but the difference was not significant. When the Q3C concentration was higher than (equal to) 1 μM, the ALP content increased significantly.

[0115] The above results show that Q3C concentrations of 1μM, 2.5μM, and 5μM can promote the proliferation and differentiation of MC3T3-E1 osteoblasts.

[0116] At the same time, the effect of quercetin derivative Q3R on the ALP level of MC3T3-E1 cells was detected. The specific process is as follows:

[0117] MC3T3-E1 cells in the logarithmic growth phase were evenly inoculated in a 24-well plate. When the cell fusion rate reached more than 70%, the old culture medium was discarded and the cells were induced to differentiate for 14 days with osteogenic differentiation medium (Induce group, containing 50 μg / mL ascorbic acid and 10 mM β-glycerophosphate) and osteogenic differentiation medium containing different concentrations of Q3R (0.25 μM, 0.5 μM, 1 μM, 2.5 μM, 5 μM). The cells in each group were collected and the ALP level was detected. During the differentiation process, the osteogenic differentiation medium was replaced every 2 to 3 days. The results are shown in Tables 6 and Figure 3 shown.

[0118] Table 6 Effect of Q3R on ALP content in MC3T3-E1 cells (n=5, mean±SD)

[0119] Group ALP content (U / L) Control Group 67.740±5.565 Induce Group <![CDATA[109.746±1.454 ### ]]> 0.25μM 110.841±2.099 0.5μM 114.377±5.571 1μM 115.203±3.457 2.5μM 110.488±6.942 5μM 109.883±5.699

[0120] From Table 6 and Figure 3 The results show that compared with the Control group, the ALP content in the Induce group increased significantly, indicating that MC3T3-E1 cells were successfully induced. Compared with the Induce group, the ALP content in each Q3R group increased slightly, but the difference was not significant, indicating that within the concentration range selected in the experiment, Q3R had no significant effect on osteoblast differentiation.

[0121] The above is only an embodiment of the present application and is not intended to limit the present application. For those skilled in the art, the present application may have various changes and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application should be included in the scope of the claims of the present application.

Claims

1. Use of quercetin-3-caffeoylrobin glycoside as an active ingredient in the preparation of a product for preventing and / or treating osteoporosis.

2. The use according to claim 1, characterized in that: The active ingredient in the product is selected from at least one of the following: A), quercetin-3-caffeoylrobin glycoside having the structural formula shown in formula (I): B) a pharmaceutically acceptable salt of quercetin-3-caffeoylrobin glycoside as described in A); C), a pharmaceutically acceptable carrier loaded with quercetin-3-caffeoylrobin glycoside as described in A).

3. The use according to claim 2, characterized in that: The pharmaceutically acceptable salt is selected from any one or more of its hydrochloride, sulfate, acetate, methanesulfonate, succinate, fumarate, citrate, malate or organic amine salt; And / or, the pharmaceutically acceptable carrier includes an inorganic carrier or a biological carrier.

4. The use according to claim 1, characterized in that: The quercetin-3-caffeoylrobin glycoside is derived from animal and plant extraction, chemical synthesis or microbial fermentation.

5. The use according to claim 1, characterized in that: The product is selected from medicines, foods, functional foods or special medical foods.

6. The use according to claim 1, characterized in that: The content of active ingredients in the product is 0.01%-99.99%, wherein the % refers to the mass volume percentage (w / v) or mass percentage (w / w).

7. The use according to claim 1, characterized in that: The product is used for promoting bone formation and / or improving the decrease in bone dry weight, bone calcium content and / or bone density caused by osteoporosis.

8. The use according to claim 1, characterized in that: The quercetin-3-caffeoylrobin glycoside in the product can also be prepared into a compound preparation with ingredients having the same activity or other activities, and the other active ingredients are selected from ingredients with anti-inflammatory, antioxidant, antibacterial, antiviral, antitumor, immunomodulatory and / or cardiovascular protective effects.

9. Use of quercetin-3-caffeoylrobin glycoside as an active ingredient in the preparation of an agent for promoting the proliferation and differentiation of osteoblasts, and increasing the bone dry weight, bone calcium content and / or bone density of animals.

10. A drug for preventing and treating osteoporosis, characterized in that: The invention comprises an active ingredient containing quercetin-3-caffeoylrobin glycoside and at least one pharmaceutically acceptable auxiliary material.

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