A lignan composition for preventing and treating osteoporosis and use thereof

Abstract

The present application provides a kind of lignan composition for resisting osteoporosis and its application, the composition is composed of pinoresinol diglucoside, dehydrodiconiferyl alcohol-4, γ'-bis-O-β-D-glucopyranoside, pinoresinol monoglucoside component;The mass ratio of pinoresinol diglucoside: dehydrodiconiferyl alcohol-4, γ'-bis-O-β-D-glucopyranoside: pinoresinol monoglucoside is 1:0.1-0.3:0.2-0.4.The lignan composition provided by the present application is stable in quality, high in safety, suitable for long-term taking, and has good effect on treating osteoporosis.The composition can be used for senile osteoporosis, and has more significant effect on inhibiting bone density reduction in ovary-removed female rat osteoporosis model.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of traditional Chinese medicine, and particularly relates to a lignan composition for resisting osteoporosis and application thereof. BACKGROUND

[0002] Osteoporosis (OP) is a systemic disease characterized by decreased bone mass and damaged bone microstructure, leading to increased bone fragility and fractures. The World Health Organization defines osteoporosis as a progressive systemic skeletal disease characterized by low bone mass and microarchitectural deterioration of bone tissue, with a consequent increase in bone fragility and susceptibility to fracture. The main pathological feature is the damage to the microstructure of bone mass, and the thinning of bone trabeculae, the widening of the gap and the decrease in the number are the main manifestations.

[0003] At present, the main clinical anti-osteoporosis drugs include estrogen, calcitonin, bisphosphonate and macromolecular monoclonal antibody, etc. Long-term use of estrogen can increase the incidence of breast cancer and endometrial hyperplasia; bisphosphonate is a first-line anti-osteoporosis drug, which can specifically bind to hydroxyapatite in bone mass, inhibit the activity of osteoclasts, and thus inhibit bone resorption. These drugs are used for treating osteoporosis, especially the osteoporosis characterized by decreased bone mass and damaged bone structure, leading to increased bone fragility and fracture rate.

[0004] The active ingredient for resisting osteoporosis is mainly lignan, including compounds such as oleuropein, syringaresinol diglucoside and pinoresinol diglucoside. Studies have shown that lignan has a certain effect on treating osteoporosis, and pinoresinol diglucoside has the effects of resisting osteoporosis and reducing blood pressure.

[0005] The Chinese invention patent CN105055381B discloses a pharmaceutical application of lignan compounds and a pharmaceutical composition. The lignan compound in the invention is a natural compound extracted and treated from the rhizome of Acorus gramineus. Research shows that the lignan compound can obviously inhibit the process of bone marrow-derived mononuclear / macrophage cells differentiating into osteoclasts and the formation of bone resorption pits, has no obvious cytotoxic effect on bone marrow-derived macrophage cells, and can be used for preparing a drug for treating or preventing / prophylactically treating osteoporosis. The inventor of the present invention discloses a eucommia extract and its application in treating osteoporosis in the patent CN111686147A. The patent discloses a eucommia extract which comprises, by weight fraction, 20-50 parts of a eucommia petroleum ether extract and 10-30 parts of a eucommia ethanol extract. The patent uses different extraction methods to extract effective components of eucommia in the implementation process, so that the effective components of the drug can be improved to different degrees. Since the solubility of different effective components in solvents is different, the content of the effective components can be effectively adjusted. The extraction method adopted by the invention is simple and easy to implement, has a significant effect in treating osteoporosis, and the composition is extremely safe for long-term use. However, the active ingredients are still not completely clear, the quality control is difficult, and the stability between batches is not easy to monitor.

[0006] Therefore, there is an urgent need for a lignan composition with stable quality, high safety and clear active ingredients, which has a better anti-osteoporosis effect. SUMMARY

[0007] The present application provides a lignan composition for preventing and treating osteoporosis and its application. The lignan composition comprises pinoresinol diglucoside, dehydrodiconiferyl alcohol-4, gamma'-bis-O-beta-D-glucopyranoside and pinoresinol monoglucoside, and has a more significant effect of preventing and treating osteoporosis.

[0008] To achieve the above object, the technical scheme adopted by the present application is as follows:

[0009] The present application provides a lignan composition for preventing and treating osteoporosis, which is composed of pinoresinol diglucoside, dehydrodiconiferyl alcohol-4, gamma'-bis-O-beta-D-glucopyranoside and pinoresinol monoglucoside.

[0010] Preferably, the mass ratio of pinoresinol diglucoside:dehydrodiconiferyl alcohol-4, gamma'-bis-O-beta-D-glucopyranoside:pinoresinol monoglucoside is 1:0.1-0.3:0.2-0.4.

[0011] Further preferably, the mass ratio of said pinoresinol diglucoside: dehydrodiconiferyl alcohol-4, γ'-bis-O-β-D-glucopyranoside: pinoresinol monoglucoside is 1:0.15-0.25:0.2-0.4.

[0012] The present application also provides a preparation method of the above lignan composition, which comprises mixing pinoresinol diglucoside, dehydrodiconiferyl alcohol-4, γ'-bis-O-β-D-glucopyranoside and pinoresinol monoglucoside to obtain the lignan composition.

[0013] The present application also provides use of the above lignan composition in preparation of a drug for preventing or treating osteoporosis.

[0014] Preferably, the osteoporosis includes postmenopausal osteoporosis and senile osteoporosis.

[0015] The present application also provides an anti-osteoporosis drug, which comprises the above lignan composition and a pharmaceutically acceptable carrier.

[0016] Preferably, the lignan composition accounts for 0.1-70% of the weight of the drug.

[0017] Further preferably, the lignan composition accounts for 0.1-50% of the weight of the drug.

[0018] Preferably, the lignan composition is used at a daily dosage of 1.0-10 mg / kg body weight.

[0019] Preferably, the dosage is an oral dosage for a human.

[0020] Further preferably, the lignan composition is used at a daily dosage of 2.0-8 mg / kg body weight.

[0021] Preferably, the dosage form of the drug includes tablets, capsules, powders, syrups, adhesives, suspensions, injections, capsules, granules.

[0022] Compared with the prior art, the present application has the following beneficial effects:

[0023] The lignan composition of the present application has a more significant therapeutic effect on senile osteoporosis and ovariectomized female rat osteoporosis, can significantly increase bone density, enhance the number of trabeculae and reduce the separation degree of trabeculae, has good safety, is suitable for long-term use, and has clear composition and controllable quality, thus meeting the needs of clinical application. BRIEF DESCRIPTION OF DRAWINGS

[0024] Figure 1The μCT scanning bone trabecular structure chart of the osteoporosis rat model in Example 1 is shown in the following table: A: sham operation group; B: model control group; C: positive control group; D: A group; E: B group; F: C group; G: D group; H: E group; I: F group; J: G group; K: H group; L: I group. DETAILED DESCRIPTION

[0025] The following non-limiting examples can make those skilled in the art more fully understand the present application, but do not limit the present application in any way. The following content is only an exemplary description of the scope of the present application, and those skilled in the art can make various changes and modifications to the application disclosed herein, which should also be within the scope of the present application.

[0026] It is worth noting that the raw materials used in the present application are ordinary commercially available products, and their sources are not specifically limited. The sources of the following raw materials are exemplary:

[0027] The CAS number of alendronate sodium is 121268-17-5;

[0028] The CAS number of pinoresinol di-glucoside is 63902-38-5;

[0029] The CAS number of pinoresinol mono-glucoside is 69251-96-3;

[0030] The CAS number of dehydrodiconiferyl alcohol 4, γ'-di-O-β-D-glucopyranoside is 109792-90-7.

[0031] In the present application, the structural formula of pinoresinol di-glucoside is:

[0032]

[0033] The structural formula of pinoresinol mono-glucoside is:

[0034]

[0035] The structural formula of dehydrodiconiferyl alcohol 4, γ'-di-O-β-D-glucopyranoside (English name: dehydrodiconiferyl alcohol 4, γ'-di-O-β-D-glucopyranoside) is:

[0036]

[0037] Example 1 Pharmacodynamic study of lignan composition

[0038] 1. Experimental animals

[0039] SPF level female SD rats 160, rat age 7-8 weeks, body weight 200±10g.

[0040] 2, the establishment of osteoporosis model

[0041] SPF level female SD rats, 7-8 weeks old, randomly selected 10 as the sham operation group, through intraperitoneal injection of 4% chloral hydrate anesthesia, except that the sham operation group only abdominal opening without removing the ovary, the remaining 150 all were bilateral ovariectomy. After operation, intraperitoneal injection of 800000 IU / kg penicillin potassium, 1 times / d continuous administration for 7 days, to prevent postoperative infection. In the 5th week, the bone mineral density (BMD) of the lumbar vertebrae and femur of each rat was determined by small animal dual energy X-ray bone density body composition analyzer (iNSiGHT VET DX), and 40 rats with no significant decrease in bone mineral density were removed. The remaining 110 SD rats (10 rats in the sham operation group were not included) were randomly divided into groups, 10 rats in each group, and fed with ordinary complete feed.

[0042] 3, grouping and dosing

[0043] Sham operation group: normal diet, n=10;

[0044] Model control group: normal diet, n=10;

[0045] Positive control group: alendronate sodium, oral gavage once a week, the dose is 6mg / kg, n=10;

[0046] Group A (pinoresinol diglucoside, 70mg / kg, n=10);

[0047] Group B (dehydrodiplopyranol-4, γ'-bis-O-β-D-glucopyranoside, 70mg / kg, n=10);

[0048] Group C (pinoresinol monoglucoside, 70mg / kg, n=10);

[0049] Group D (pinoresinol diglucoside: dehydrodiplopyranol-4, γ'-bis-O-β-D-glucopyranoside: pinoresinol monoglucoside = 1:0.1:0.4, 65mg / kg, n=10);

[0050] Group E (pinoresinol diglucoside: dehydrodiplopyranol-4, γ'-bis-O-β-D-glucopyranoside: pinoresinol monoglucoside = 1:0.3:0.2, 65mg / kg, n=10);

[0051] Group F (pinoresinol diglucoside: dehydrodiconiferyl alcohol-4, γ'-bis-O-β-D-glucopyranoside: pinoresinol monoglucoside = 1:0.15:0.2, 65 mg / kg, n = 10);

[0052] Group G (pinoresinol diglucoside: dehydrodiconiferyl alcohol-4, γ'-bis-O-β-D-glucopyranoside: pinoresinol monoglucoside = 1:0.25:0.4, 65 mg / kg, n = 10);

[0053] Group H (pinoresinol diglucoside: dehydrodiconiferyl alcohol-4, γ'-bis-O-β-D-glucopyranoside: pinoresinol monoglucoside = 1:0.4:1, 65 mg / kg, n = 10);

[0054] Group I (pinoresinol diglucoside: dehydrodiconiferyl alcohol-4, γ'-bis-O-β-D-glucopyranoside: pinoresinol monoglucoside = 1:0.5:0.5, 65 mg / kg, n = 10);

[0055] All groups were mixed in the proportions of each group to obtain the corresponding composition. The drugs in each group were orally administered by gavage, once a day for 16 consecutive weeks, with the same administration time each day.

[0056] 4. Evaluation of indicators such as bone mineral density and trabecular bone structure:

[0057] After the 16-week administration period, the bone mineral density of the femur and lumbar vertebrae of the rats was measured under anesthesia; the blood was taken from the orbit to measure the serum biochemical indicators; the rats were sacrificed and the femur was removed to measure the trabecular bone structure using μCT. The efficacy of the composition against osteoporosis was evaluated by the above indicators.

[0058] The instrument for measuring the bone mineral density of the femur and lumbar vertebrae of the rats was the iNSiGHT VET DXA type dual-energy X-ray bone densitometer produced by OsteoSys Company of South Korea; the reagent kits for measuring the serum biochemical indicators ALP, blood phosphorus and blood calcium were purchased from Nanjing Jiancheng Biological Engineering Institute, with item numbers A059-2-2, C006-1-1 and C004-2-1 respectively; the instrument for measuring the trabecular bone structure was Skyscan 1174 X-Ray Microtomograph (μCT) from Bruker Company of Belgium; the sample scanning parameters were as follows: voltage 50 kV, current 800 μA, scanning resolution 12 μm, field size 1304x1024, and the femur of the rats was scanned.

[0059] 5. Results

[0060] Table 1 Results of bone mineral density measurement of the femur and lumbar vertebrae of rats in each group after 16 weeks of administration (χ ± SD, n = 10)

[0061]

[0062]

[0063] Note: * means P < 0.05 compared with the model control group; ** means P < 0.01 compared with the model control group; # P < 0.05 compared with the sham operation group; ▲ P < 0.05 compared with the positive control group.

[0064] As can be seen from the results in Table 1, after ovariectomy, the lumbar spine bone density and femur bone density of the rats were significantly lower than those of the sham operation group (P < 0.01), indicating that the rat osteoporosis model was successfully established.

[0065] After 16 weeks of administration, the lumbar spine bone density and femur bone density of the rats in the positive group were increased to 0.183 ± 0.012 and 0.268 ± 0.011, respectively, which were higher than the lumbar spine bone density and femur bone density of the model group, and the difference was statistically significant (P < 0.05), indicating that alendronate sodium can reverse the decrease in bone density of the model rats.

[0066] In groups A, B and C, pinocembrin diglucoside, dehydrodiconiferyl alcohol-4, γ'-bis-O-β-D-glucopyranoside and pinocembrin monoglucoside were used alone, respectively. After 16 weeks of administration, the lumbar spine and femur bone densities of the rats were increased to a certain extent compared with the model control group, indicating that the three lignan components used alone had a certain anti-osteoporosis effect. Compared with the positive drug alendronate sodium, the three compounds increased the lumbar spine and femur bone densities of the rats to a similar extent. Although the lumbar spine and femur bone densities of the rats in group C were slightly higher than those of the positive control group, no statistically significant difference was observed. In groups D, E, F and G, the three lignan glycoside components pinocembrin diglucoside, dehydrodiconiferyl alcohol-4, γ'-bis-O-β-D-glucopyranoside and pinocembrin monoglucoside were combined in a certain proportion to form compositions, and the combination proportions were 1:0.1:0.4, 1:0.3:0.2, 1:0.15:0.2 and 1:0.25:0.4, respectively. Under the condition of a dose of 65 mg / kg, the three lignan components combined in a certain proportion could form a more effective synergistic effect against osteoporosis, and the mass ratio of pinocembrin diglucoside:dehydrodiconiferyl alcohol-4, γ'-bis-O-β-D-glucopyranoside:pinocembrin monoglucoside was 1:0.15-0.25:0.2-0.4, which had the most effective effect against osteoporosis.

[0067] To further verify the ratio range, the present experiment added H group and I group, which were given the three lignan components in the ratio of 1:0.4:1 and 1:0.5:0.5, respectively. The lumbar spine bone density and femur bone density data of the rats after 16 weeks of administration are shown in Table 1. It can be seen that the lumbar spine bone density and femur bone density of the rats in the H group and the I group were significantly higher than the corresponding bone density of the model control group (except for the femur bone density of the H group, which showed no statistical difference compared with the model control group), indicating that the composition had an anti-osteoporosis effect, but the anti-osteoporosis effect of the composition of the two groups was not as good as alendronate sodium compared with the positive control group. Although there was no statistical difference, the bone density was lower than that of the positive control group, which did not show a good treatment advantage, and the anti-bone density reduction effect of the H group and the I group was significantly weakened compared with the D group, the E group, the F group, and the G group.

[0068] The composition of lignans significantly improved the bone density of the ovariectomized rats in the ratio range of pinoresinol diglucoside: dehydrodiconiferyl alcohol-4, γ'-bis-O-β-D-glucopyranoside: pinoresinol monoglucoside = 1:0.1-0.3:0.2-0.4, and the therapeutic effect was more significant compared with the positive drug alendronate sodium.

[0069] Table 2 Changes in femur trabecular structure of rats after 16 weeks of administration (χ±SD, n=10)

[0070]

[0071]

[0072] Note: *P<0.05 compared with the model group; **P<0.01 compared with the model group; ## P<0.01 compared with the sham operation group; ★ P<0.05 compared with the positive control group.

[0073] After ovariectomy, the femur microstructure of the rats was significantly damaged, the trabecular bone number was significantly reduced, the trabecular bone separation degree was increased, and the bone volume fraction was significantly decreased, as shown in Table 2. The bone volume fraction BV / TV of the model control group was reduced to 12.5±2.0%, which was significantly lower than that of the sham operation group (36.7±7.5%), the trabecular bone number Tb.N of the model group was reduced to 1.17±0.28 (1 / mm), which was significantly lower than that of the sham operation group (4.04±0.69 (1 / mm)), and the trabecular bone separation degree of the model group was increased to 0.71±0.14 (mm), which was significantly higher than that of the sham operation group (0.22±0.05 (mm)), P<0.01, indicating that the rat osteoporosis model in the present experiment was successfully established.

[0074] After 16 weeks of administration, the bone volume fraction BV / TV and the trabecular bone number Tb.N (1 / mm) of the positive group rats administered with sodium alendronate were increased to 17.1 ± 7.2% and 2.14 ± 0.37 (1 / mm), respectively. The former was significantly increased, but due to the large individual differences of the rats in the group, it did not reach a statistical difference compared with the model control group, while Tb.N had a statistical difference compared with the model control group (P < 0.05), indicating that sodium alendronate can significantly increase the number of trabecular bone within a unit size; the trabecular bone separation degree is an important parameter for measuring the health of bone microstructure, and after 16 weeks of administration of sodium alendronate, the Tb.Sp of the positive control group was decreased from 0.71 ± 0.14 of the model control group to 0.43 ± 0.10, and the difference had a statistically significant difference (P < 0.01).

[0075] After 16 weeks of continuous administration, the bone volume fraction and the trabecular bone number of the rats in groups A, B and C corresponding to the three lignan components pinoresinol diglucoside, dehydrodiconiferyl alcohol-4, γ'-bis-O-β-D-glucopyranoside and pinoresinol monoglucoside were significantly increased, and the increasing effect was equivalent to that of the positive drug sodium alendronate, and there were statistical differences compared with the model control group, indicating that the three lignan components had the activity of protecting the trabecular bone microstructure; the determination results of the bone separation degree showed that groups A, B and C were significantly lower than the model control group, and the difference was extremely significant (P < 0.01), further indicating that the three components had the effect of resisting the damage of the trabecular bone. After 16 weeks of administration, the bone volume fraction and the trabecular bone number of the rats in groups D, E, F and G were significantly higher than those of the model control group, and the difference had statistical significance and biological significance compared with the model control group (P < 0.01); as shown in Table 2, the trabecular bone separation degree of the rats in groups D, E, F and G was significantly lower than that of the model control group, and the difference had statistical significance. The bone volume fraction and the trabecular bone number of the rats in groups H and I were significantly higher than those of the model control group, and the trabecular bone separation degree was lower than that of the model control group, and the results had statistical differences. However, compared with groups D, E, F and G, the effect of groups H and I on increasing the bone volume fraction and the trabecular bone number and reducing the bone separation degree was not as significant as that of groups D, E, F and G, indicating that the anti-osteoporosis effect of groups H and I was not as significant as that of groups D, E, F and G, indicating that the anti-osteoporosis effect of the composition in different combination ratios was different. The results showed that compared with the positive drug sodium alendronate which has been marketed, the composition composed of pinoresinol diglucoside, dehydrodiconiferyl alcohol-4, γ'-bis-O-β-D-glucopyranoside and pinoresinol monoglucoside in a mass ratio of 1:0.1-0.3:0.2-0.4 had the most significant effect on increasing the bone volume fraction and the trabecular bone number and reducing the trabecular bone separation degree.

[0076] Table 3 Determination results of part of serum biochemical indexes of rats after 16 weeks of administration

[0077]

[0078]

[0079] *P < 0.05 compared to the model group, **P < 0.01 compared to the model group. # Compared with the sham surgery group, P < 0.05.

[0080] As shown in Table 3, after 16 weeks of administration, compared with the model control group, the activity of alkaline phosphatase (ALP) in rats of the positive control group, groups A, B, and C, and groups D, E, F, G, H, and I containing lignan compositions was significantly reduced. This indicates that both the test drug and the positive control drug, alendronate sodium, can reduce ALP in osteoporotic rats. The most significant reductions in ALP concentration were observed in groups D, E, F, and G, suggesting that a ratio of pinoresinol diglucoside: dehydrobispineol-4,γ'-bis-O-β-D-glucopyranoside: pinoresinol monoglucoside = 1:0.1-0.3:0.2-0.4 has a more significant ALP-reducing effect. Serum phosphorus and calcium levels were measured in each group, and no significant differences were found between the groups.

[0081] The lignan composition significantly increased bone mineral density in castrated female rats, demonstrating a marked anti-osteoporosis effect. It also increased the number and thickness of trabeculae and reduced bone separation, indicating a significant therapeutic effect on osteoporosis.

[0082] Example 2: Efficacy test of anti-bone mineral density reduction in aged male rats.

[0083] 1. Laboratory animals

[0084] 180 male SPF-grade SD rats, aged 7-8 weeks and weighing (200±20)g.

[0085] 2. Establishment of an osteoporosis model

[0086] Male SD rats, aged 7-8 weeks, were selected and fed a normal diet until 18 months of age. Bone mineral density (BMD) of the lumbar vertebrae and femur was measured using a small animal dual-energy X-ray absorptiometry (iNSiGHT VET DX) system. Sixty rats without significantly reduced BMD were excluded, and the remaining 120 SD rats were randomly divided into groups of 10 each and fed a standard complete diet. Ten additional male SD rats, aged 8 weeks, were purchased before the end of the experiment to serve as a normal control group.

[0087] 3. Grouping and Dosage

[0088] Normal control group: normal diet, n = 10;

[0089] Model control group: normal diet, n = 10;

[0090] Positive control group: alendronate sodium, oral gavage once a week, dose 6 mg / kg, n = 10;

[0091] Group I (pinoresinol diglucoside, 60 mg / kg)

[0092] Group II (dehydrodiplopterol-4, γ'-bis-O-β-D-glucopyranoside, 60 mg / kg)

[0093] Group III (pinoresinol monoglucoside, 60 mg / kg)

[0094] Group IV (pinoresinol diglucoside: dehydrodiplopterol-4, γ'-bis-O-β-D-glucopyranoside: pinoresinol monoglucoside = 1:0.1:0.4, 50 mg / kg)

[0095] Group V (pinoresinol diglucoside: dehydrodiplopterol-4, γ'-bis-O-β-D-glucopyranoside: pinoresinol monoglucoside = 1:0.3:0.2, 50 mg / kg)

[0096] Group VI (pinoresinol diglucoside: dehydrodiplopterol-4, γ'-bis-O-β-D-glucopyranoside: pinoresinol monoglucoside = 1:0.15:0.2, 50 mg / kg)

[0097] Group VII (pinoresinol diglucoside: dehydrodiplopterol-4, γ'-bis-O-β-D-glucopyranoside: pinoresinol monoglucoside = 1:0.25:0.4, 50 mg / kg)

[0098] Group VIII (pinoresinol diglucoside: dehydrodiplopterol-4, γ'-bis-O-β-D-glucopyranoside: pinoresinol monoglucoside = 1:0.4:1, 50 mg / kg)

[0099] Group IX (pinoresinol diglucoside: dehydrodiplopterol-4, γ'-bis-O-β-D-glucopyranoside: pinoresinol monoglucoside = 1:0.5:0.5, 50 mg / kg)

[0100] Group X (the extract of eucommia oil with petroleum ether: the extract of eucommia with ethanol = 2:1, 100 mg / kg)

[0101] All groups were mixed in the corresponding proportions to obtain the corresponding compositions, and the drugs in each group were orally gavaged for 12 consecutive months. Once a day, the same time for each administration.

[0102] 4. Bone density measurement and results

[0103] After 12 months of administration, the bone density of the femur and lumbar vertebrae of the rats was measured under anesthesia to evaluate the anti-osteoporosis efficacy of the composition.

[0104] The instrument for measuring the bone density of the femur and lumbar vertebrae of the rats was a dual-energy X-ray absorptiometry (DXA) type iNSiGHT VET produced by OsteoSys Co., Ltd. of Korea.

[0105] Table 4. Results of bone density measurement of the femur and lumbar vertebrae of the rats after 12 months of administration (χ ± SD, n = 10)

[0106] Group Lumbar spine bone density (g / cm 2 ) Femur bone density (g / cm 2 )]]> Normal control group 0.232±0.013 0.296±0.016 Model control group 0.204 ± 0.012 ## ]] 0.250 ± 0.014 ## ]] Positive control group 0.227±0.014* 0.287±0.012* Group I 0.215±0.008* 0.260±0.017* Group II 0.208±0.010* 0.262±0.013* Group III 0.216±0.012* 0.268±0.012* Group IV 0.225 ± 0.007** ▲ ]] 0.285 ± 0.011** ▲ ]] Group V 0.227 ± 0.011** ▲ ]] 0.289 ± 0.010** ▲ ]] Group VI 0.231 ± 0.008** ▲ ]] 0.291 ± 0.012** ▲ ]] Group VII 0.229 ± 0.011** ▲ ]] 0.295 ± 0.012** ▲ ]] Group VIII 0.212±0.013* 0.265±0.011 Group IX 0.215±0.012* 0.268±0.017* Group X 0.215±0.016* 0.258±0.015*

[0107] Note: * indicates P < 0.05 compared with the model control group; ** indicates P < 0.01 compared with the model control group; # indicates P < 0.05 compared with the normal control group; ▲ indicates P < 0.05 compared with the positive control group.

[0108] As can be seen from Table 4, the bone density of the lumbar vertebrae and the femur of the rats was significantly lower than that of the normal control group (P < 0.01) after ovariectomy, indicating that the rat osteoporosis model was successfully established.

[0109] After 12 months of continuous administration, the lumbar spine bone density and femur bone density of the positive group rats given alendronate intervention increased to 0.227±0.014 and 0.287±0.012, respectively, which were higher than the lumbar spine bone density and femur bone density of the model group, and the difference was statistically significant (P<0.05), indicating that alendronate can reverse the decrease of bone density in the model rats. In group I, group II and group III, robinin di-glucoside, dehydrodiplopterol-4, γ'-bis-O-β-D-glucopyranoside and robinin mono-glucoside were used alone, respectively. After 12 months of administration, the lumbar spine and femur bone density of the rats in each group was increased to a certain extent compared with the model control group, indicating that the three lignan components used alone have a certain anti-osteoporosis effect. Compared with the positive drug alendronate, the three compounds did not improve the lumbar spine and femur bone density of the rats as much as the positive control drug, but there was no statistically significant difference. In group IV, group V, group VI and group VII, the drugs given were the three lignan glycoside components of robinin di-glucoside, dehydrodiplopterol-4, γ'-bis-O-β-D-glucopyranoside and robinin mono-glucoside, which were combined in a certain proportion. The combination proportions were 1:0.1:0.4, 1:0.3:0.2, 1:0.15:0.2 and 1:0.25:0.4, respectively. Under the condition of a dose of 50 mg / kg, the three lignan components combined in a certain proportion can form a more effective anti-osteoporosis synergistic effect, and the mass ratio of robinin di-glucoside:dehydrodiplopterol-4, γ'-bis-O-β-D-glucopyranoside:robinin mono-glucoside is 1:0.1-0.3:0.2-0.4, which has the most effective anti-osteoporosis effect.

[0110] To further verify the ratio range, the present experiment added group VIII and group IX, which were given the three lignan components in the ratio of 1:0.4:1 and 1:0.5:0.5, respectively. After 12 months of continuous administration, the lumbar spine bone density and femur bone density of the rats in group VIII and group IX were as shown in Table 4. It can be seen that the lumbar spine bone density and femur bone density of the rats in group VIII and group IX were significantly higher than the corresponding bone density of the model control group (except that the femur bone density of group VIII did not show statistical difference compared with the model control group), indicating that the composition had an anti-osteoporosis effect. However, compared with the positive control group, the anti-osteoporosis effect of the composition in group VIII and group IX was not as good as that of alendronate sodium, although there was no statistical difference, but the bone density was lower than that of the positive control group, which did not show a good treatment advantage, and compared with group IV, group V, group VI and group VII, the anti-bone density reduction effect of group VIII and group IX was significantly weakened. In the present study, group X was also set up, and the tested drug used was a composition of eucommia ulmoides oil ether extract and eucommia ulmoides ethanol extract in a mass ratio of 2:1. The results showed that after 12 months of continuous gavage, the lumbar spine bone density and femur bone density of the rats in group X were 0.215±0.016 and 0.258±0.015, respectively. Although the two were significantly improved compared with the corresponding bone density of the model control group, they had biological significance and statistical significance, but the anti-osteoporosis effect was not as good as that of the lignan composition provided in the present application, even when the administration dose was significantly increased to 100 mg / kg, the therapeutic effect was still weaker than that of the lignan composition provided in the present application.

[0111] In summary, the lignan composition significantly improved the bone density of the ovariectomized rats in the ratio range of pinoresinol diglucoside: dehydrodiplopterol-4, γ'-bis-O-β-D-glucopyranoside: pinoresinol monoglucoside = 1:0.1-0.3:0.2-0.4, and the therapeutic effect was more significant compared with the positive drug alendronate sodium.

[0112] The above lignan composition can be made into tablets, capsules or granules. The specific method is as follows: lignan composition 10 g, microcrystalline cellulose 1.8 g, magnesium stearate 0.2 g, mix uniformly, granulate, and then make tablets, capsules or granules.

[0113] The above lignan composition can be made into capsules. The specific method is as follows: lignan composition 10 g, pre-crosslinked starch 3.8 g, mix uniformly, granulate, and then fill the capsules.

[0114] Finally, it should be noted that the above content is only used to illustrate the technical solutions of the present application, and is not a limitation on the protection scope of the present application. Simple modifications or equivalent replacements of the technical solutions of the present application made by those skilled in the art do not deviate from the essence and scope of the technical solutions of the present application.

Claims

1. An anti-osteoporotic lignan composition, characterized by: The lignan composition is composed of pinoresinol diglucoside, dehydrodiconiferyl alcohol-4, γ'-bis-O-β-D-glucopyranoside and pinoresinol monoglucoside in a mass ratio of 1:0.1-0.3:0.2-0.

4.

2. The lignan composition of claim 1, wherein: The mass ratio of the pinoresinol diglucoside:dehydrodiconiferyl alcohol-4, γ'-bis-O-β-D-glucopyranoside:pinoresinol monoglucoside is 1:0.15-0.25:0.2-0.

4.

3. A method of preparing a lignan composition according to any one of claims 1-2, characterized by: The preparation method comprises mixing pinoresinol diglucoside, dehydrodiconiferyl alcohol-4, γ'-bis-O-β-D-glucopyranoside and pinoresinol monoglucoside to obtain the lignan composition.

4. Use of the lignan composition of any one of claims 1-2 or the lignan composition prepared by the preparation method of claim 3 in the preparation of a drug for preventing or treating osteoporosis.

5. Use according to claim 4, characterized in that: The osteoporosis is postmenopausal osteoporosis or senile osteoporosis.

6. An agent for preventing or treating osteoporosis, characterized by comprising the compound of the formula (I) or a pharmaceutically acceptable salt thereof. The lignan composition comprises the lignan composition of any one of claims 1-2 and a pharmaceutically acceptable carrier.

7. The medicament according to claim 6, characterized in that: The lignan composition accounts for 0.1-70% of the weight of the drug.

8. The medicament according to claim 6, characterized in that: The dosage form of the drug comprises tablets, capsules, powders, syrups, adhesives, suspensions, injections and granules.

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