阅读说明：本技术 一种含松脂醇二葡萄糖苷的组合物及其制备方法和应用 (Composition containing pinoresinol diglucoside and preparation method and application thereof ) 是由 肖军平 陈梁 李小锋 刘厚权 张梅红 胡吉忠 夏淑英 毛金娣 林飞英 于 2021-06-30 设计创作，主要内容包括：本发明属于药物制备技术领域,具体涉及一种含松脂醇二葡萄糖苷的组合物及其制备方法和应用。含松脂醇二葡萄糖苷的组合物,包括以下组分：松脂醇二葡萄糖苷、丁香脂素、木耳多糖、γ-氨基丁酸、甘露三糖、佛手苷内酯。该组合物具有较好的治疗治疗骨质疏松症的功效,制备方法简单,毒副作用小、成本低。(The invention belongs to the technical field of medicine preparation, and particularly relates to a composition containing pinoresinol diglucoside, and a preparation method and application thereof. A composition containing pinoresinol diglucoside comprises the following components: pinoresinol diglucoside, syringaresinol, Auricularia polysaccharide, gamma-aminobutyric acid, manninotriose and bergamotol lactone. The composition has good therapeutic effect on osteoporosis, and has the advantages of simple preparation method, low adverse side effect, and low cost.)

1. A composition containing pinoresinol diglucoside comprises the following components:

pinoresinol diglucoside, syringaresinol, Auricularia polysaccharide, gamma-aminobutyric acid, manninotriose and bergamotol lactone.

2. The pinoresinol diglucoside-containing composition according to claim 1, comprising the following components in parts by weight:

10-20 parts of pinoresinol diglucoside, 1-5 parts of syringaresinol, 5-10 parts of agaric polysaccharide, 1-5 parts of gamma-aminobutyric acid, 1-10 parts of manninotriose and 5-15 parts of bergamoolide.

3. The pinoresinol diglucoside-containing composition according to claim 1, comprising the following components in parts by weight:

12-16 parts of pinoresinol diglucoside, 3-5 parts of syringaresinol, 7-9 parts of agaric polysaccharide, 2-4 parts of gamma-aminobutyric acid, 4-8 parts of manninotriose and 10-12 parts of bergamoolide.

4. The pinoresinol diglucoside-containing composition according to any one of claims 1 to 3, wherein the agaric polysaccharide is prepared by:

(1) cleaning Auricularia, soaking, homogenizing, extracting in distilled water, and filtering to obtain primary extractive solution and residue;

(2) adding distilled water into the filter residue, and performing ultrasonic extraction to obtain a secondary extracting solution;

(3) mixing the primary and secondary extractive solutions, concentrating under reduced pressure, and precipitating with ethanol; preparing alcohol precipitate;

(4) dissolving the alcohol precipitate in water, treating the alcohol precipitate by sephadexG-100 gel column, eluting by using ethyl acetate, collecting ethyl acetate eluent, eluting by using distilled water, and collecting distilled water eluent; adding distilled water into ethyl acetate eluate for extraction, mixing the separated water phase with distilled water eluate, and concentrating to obtain Auricularia polysaccharide.

5. The pinoresinol diglucoside-containing composition according to claim 4, wherein in the step (1), the extraction is performed for 1-2h by boiling extraction after 30-60min at 60-80 ℃; in the step (2), the power of the ultrasound is 80-100W; the extraction time is 30-60 min.

6. A process for the preparation of a pinoresinol diglucoside-containing composition according to any one of claims 1 to 5, comprising the steps of:

dissolving gamma-aminobutyric acid in distilled water, adding solubilizer, and dispersing pinoresinol diglucoside, syringaresinol, Auricularia polysaccharide, mannotriose, and bergamoolide in distilled water to obtain mixture.

7. The method of claim 6, wherein the solubilizer is any one or more of propylene glycol, polyethylene glycol laurate and polyethylene glycol glyceryl stearate.

8. The method of claim 7, wherein the solubilizer is a mixture of propylene glycol and polyethylene glycol laurate, wherein the propylene glycol and polyethylene glycol laurate are present in a mass ratio of 1-3: 1; the relative molecular weight of the polyethylene glycol laurate is 600-1000; the mass concentration of the solubilizer in distilled water is 1-5%.

9. Use of a composition comprising pinoresinol diglucoside according to any one of claims 1 to 5 or a composition comprising pinoresinol diglucoside according to any one of claims 6 to 8, which is prepared by the method for preparing the composition, for preparing a medicament for preventing and treating osteoporosis.

10. A pharmaceutical preparation comprising the composition containing pinoresinol diglucoside according to any one of claims 1 to 5 or the composition containing pinoresinol diglucoside according to any one of claims 6 to 8, and pharmaceutically usual excipients.

Technical Field

The invention belongs to the technical field of medicine preparation, and particularly relates to a composition containing pinoresinol diglucoside, and a preparation method and application thereof.

Background

Osteoporosis (osteoporotis) is a systemic bone disease in which bone fracture is easily caused by a decrease in bone density and bone mass due to various causes, a destruction of bone microarchitecture, and an increase in bone fragility. Osteoporosis is divided into primary and secondary categories. Primary osteoporosis is further classified into postmenopausal osteoporosis, senile osteoporosis and idiopathic osteoporosis.

There are a variety of methods for treating osteoporosis in the prior art. For example, the chinese patent application CN1583069A discloses a medicine for treating osteoporosis, which is composed of the following raw materials in parts by weight: 8-12 parts of drynaria rhizome, 4-7 parts of eucommia bark, 4-7 parts of parasitic loranthus, 2-4 parts of cibotium rhizome, 3-6 parts of ligusticum wallichii, 3-5 parts of angelica sinensis, 4-6 parts of safflower, 3-5 parts of bighead atractylodes rhizome and 6-10 parts of medicinal cyathula root. The medicine is prepared by selecting medicines according to the theory of 'kidney governs bone and produces marrow' in the traditional Chinese medicine, mainly tonifying liver and kidney, giving consideration to treatment methods of tonifying spleen and stomach, promoting blood circulation and removing blood stasis, and the like. The clinical and animal in-vivo and in-vitro experimental studies show that the medicine for treating osteoporosis can fundamentally adjust bone metabolism and convert the bone metabolism to a normal balance direction.

Chinese patent application CN1079160A discloses a Chinese herbal medicine which is very effective in treating osteoporosis broken bone and consists of 18 Chinese herbal medicines of agilawood, saffron, elecampane, ligusticum wallichii, cassia twig, angelica dahurica, pseudo-ginseng, teasel root, ground beeltle, rhizoma drynariae, processed nux vomica, achyranthes bidentata, cucumber seed, chicken bone, rheum officinale, native copper, borneol and dragon's blood.

Chinese patent application CN111714577A discloses a medicine "zhengyuan gutai" for preventing and treating osteoporosis and its preparation method. It is a micromolecule traditional Chinese medicine which is prepared by pure natural medicinal materials such as cornus, gordon euryale seed, cherokee rose, yam, goat bone, ox tendon, deer tendon and the like by using biotechnology and takes compound amino acid as main material; has effects in nourishing yin, warming yang, removing blood stasis, dredging meridians, invigorating kidney, and strengthening bone.

Most of the existing methods for treating osteoporosis have limited treatment effect, and meanwhile, a part of Chinese herbal medicine formula has more components, wherein a plurality of impurities exist, so that the medicine effect is influenced, the utilization rate of the medicine is greatly reduced, and on the other hand, side effects are possibly generated to be not beneficial to the health.

The invention aims to research a composition containing pinoresinol diglucoside for treating osteoporosis, which has the advantages of small side effect, low cost and good effect, and can effectively solve the problems.

Disclosure of Invention

In order to overcome the technical problems, the invention provides a composition containing pinoresinol diglucoside and a preparation method and application thereof; the composition has good therapeutic effect on osteoporosis, and has the advantages of simple preparation method, low adverse side effect, and low cost.

In order to achieve the above purpose, the technical scheme provided by the invention is as follows:

a composition containing pinoresinol diglucoside comprises the following components:

pinoresinol diglucoside, syringaresinol, Auricularia polysaccharide, gamma-aminobutyric acid, manninotriose and bergamotol lactone.

Preferably, the composition comprises the following components in parts by weight:

10-20 parts of pinoresinol diglucoside, 1-5 parts of syringaresinol, 5-10 parts of agaric polysaccharide, 1-5 parts of gamma-aminobutyric acid, 1-10 parts of manninotriose and 5-15 parts of bergamoolide.

Preferably, the composition comprises the following components in parts by weight:

12-16 parts of pinoresinol diglucoside, 3-5 parts of syringaresinol, 7-9 parts of agaric polysaccharide, 2-4 parts of gamma-aminobutyric acid, 4-8 parts of manninotriose and 10-12 parts of bergamoolide.

Preferably, the preparation method of the agaric polysaccharide comprises the following steps:

(1) cleaning Auricularia, soaking, homogenizing, extracting in distilled water, and filtering to obtain primary extractive solution and residue;

(2) adding distilled water into the filter residue, and performing ultrasonic extraction to obtain a secondary extracting solution;

(3) mixing the primary and secondary extractive solutions, concentrating under reduced pressure, and precipitating with ethanol; preparing alcohol precipitate;

(4) dissolving the alcohol precipitate in water, treating the alcohol precipitate by sephadexG-100 gel column, eluting by using ethyl acetate, collecting ethyl acetate eluent, eluting by using distilled water, and collecting distilled water eluent; adding distilled water into ethyl acetate eluate for extraction, mixing the separated water phase with distilled water eluate, and concentrating to obtain Auricularia polysaccharide.

Preferably, in step (1), the extraction is performed at 60-80 deg.C for 30-60min and boiling for 1-2 h.

Preferably, in the step (2), the power of the ultrasound is 80-100W; the extraction time is 30-60 min.

Another object of the present invention is to provide a method for preparing the composition containing pinoresinol diglucoside, comprising the steps of:

dissolving gamma-aminobutyric acid in distilled water, adding solubilizer, and dispersing pinoresinol diglucoside, syringaresinol, Auricularia polysaccharide, mannotriose, and bergamoolide in distilled water to obtain mixture.

Preferably, the solubilizer is any one or more of propylene glycol, polyethylene glycol laurate and polyethylene glycol glyceryl stearate.

Preferably, the solubilizer is a mixture of propylene glycol and polyethylene glycol laurate, wherein the mass ratio of the propylene glycol to the polyethylene glycol laurate is 1-3: 1.

Preferably, the polyethylene glycol laurate has a relative molecular weight of 600-1000.

Preferably, the mass concentration of the solubilizer in distilled water is 1-5%.

The invention also aims to provide application of the composition containing pinoresinol diglucoside in preparing a medicament for preventing and treating osteoporosis.

The invention also aims to provide a pharmaceutical preparation which comprises the composition containing pinoresinol diglucoside and pharmaceutically common auxiliary materials.

Compared with the prior art, the invention has the technical advantages that:

(1) the composition containing pinoresinol diglucoside provided by the invention can effectively prevent and treat osteoporosis.

(2) The agaric polysaccharide can promote blood circulation, has the effect of improving the immunity of the organism, can effectively promote the absorption of calcium of the organism and improve osteoporosis; meanwhile, the pinoresinol diglucoside, the agaric polysaccharide and the gamma-aminobutyric acid have good synergistic effect, and can effectively prevent and treat osteoporosis and enhance the bone density.

(3) Syringaresinol, manninotriose and bergamotol have good synergistic effect, and can effectively promote the bone strength, immunity, fatigue resistance and other effects of the pinoresinol diglucoside.

(4) SephadexG100 is used in the extraction process of the agaric polysaccharide, so that the separation of the polysaccharide from protein and other impurity small molecules can be effectively promoted, and the purification of the agaric polysaccharide is promoted.

(5) According to the invention, the agaric is extracted by combining water extraction and ultrasonic extraction, so that polysaccharide components in the agaric can be effectively extracted, and the utilization rate of the agaric is improved.

(6) When the mixed solution is prepared, the mixture of the propylene glycol and the polyethylene glycol laurate is selected as the solubilizer, so that on one hand, the dissolubility of each component in water can be effectively promoted, and on the other hand, when the auxiliary materials are added to prepare the preparation, the utilization rate of the effective components can be improved, and the effect can be effectively exerted.

Detailed Description

The present invention will be described below with reference to specific examples to make the technical aspects of the present invention easier to understand and grasp, but the present invention is not limited thereto. The experimental methods described in the following examples are all conventional methods unless otherwise specified; the reagents and materials are commercially available, unless otherwise specified.

Example 1

The composition containing pinoresinol diglucoside comprises the following components in parts by weight:

13 parts of pinoresinol diglucoside, 4 parts of syringaresinol, 8 parts of agaric polysaccharide, 3 parts of gamma-aminobutyric acid, 5 parts of manninotriose and 11 parts of bergamoolide.

The preparation method of the agaric polysaccharide comprises the following steps:

(1) cleaning Auricularia, soaking, homogenizing, extracting with distilled water at 70 deg.C for 40min, boiling for 1.5 hr, and filtering to obtain primary extractive solution and residue;

(2) adding distilled water into the filter residue, and performing ultrasonic extraction for 50min with ultrasonic power of 80W to obtain secondary extract;

(3) mixing the primary and secondary extractive solutions, concentrating under reduced pressure, and precipitating with ethanol; preparing alcohol precipitate;

(4) dissolving the alcohol precipitate in water, treating the alcohol precipitate by sephadexG-100 gel column, eluting by using ethyl acetate, collecting ethyl acetate eluent, eluting by using distilled water, and collecting distilled water eluent; adding distilled water into ethyl acetate eluate for extraction, mixing the separated water phase with distilled water eluate, and concentrating to obtain Auricularia polysaccharide.

The preparation method of the composition containing pinoresinol diglucoside comprises the following steps:

dissolving gamma-aminobutyric acid in 200 parts of distilled water, adding a mixture of 2:1 propylene glycol and polyethylene glycol laurate (M is 800) as a solubilizer, wherein the mass concentration of the solubilizer in the distilled water is 2%, and then dispersing pinoresinol diglucoside, syringaresinol, agaric polysaccharide, manninotriose and bergapten in the distilled water to prepare a mixed solution of the composition.

Example 2

The composition containing pinoresinol diglucoside comprises the following components in parts by weight:

10 parts of pinoresinol diglucoside, 1 part of syringaresinol, 5 parts of agaric polysaccharide, 1 part of gamma-aminobutyric acid, 10 parts of manninotriose and 15 parts of bergamoolide.

The preparation method of the agaric polysaccharide comprises the following steps:

(1) cleaning Auricularia, soaking, homogenizing, extracting with distilled water at 60 deg.C for 60min, boiling for 1 hr, and filtering to obtain primary extractive solution and residue;

(2) adding distilled water into the filter residue, and performing ultrasonic extraction for 30min with ultrasonic power of 100W to obtain secondary extract;

(3) mixing the primary and secondary extractive solutions, concentrating under reduced pressure, and precipitating with ethanol; preparing alcohol precipitate;

(4) dissolving the alcohol precipitate in water, treating the alcohol precipitate by sephadexG-100 gel column, eluting by using ethyl acetate, collecting ethyl acetate eluent, eluting by using distilled water, and collecting distilled water eluent; adding distilled water into ethyl acetate eluate for extraction, mixing the separated water phase with distilled water eluate, and concentrating to obtain Auricularia polysaccharide.

The preparation method of the composition containing pinoresinol diglucoside comprises the following steps:

dissolving gamma-aminobutyric acid in 200 parts of distilled water, adding a mixture of 1:1 propylene glycol and polyethylene glycol laurate (M600) as a solubilizer, wherein the mass concentration of the solubilizer in the distilled water is 5%, and dispersing pinoresinol diglucoside, syringaresinol, agaric polysaccharide, manninotriose and bergapten in the distilled water to prepare a mixed solution of the composition.

Example 3

The composition containing pinoresinol diglucoside comprises the following components in parts by weight:

20 parts of pinoresinol diglucoside, 5 parts of syringaresinol, 10 parts of agaric polysaccharide, 5 parts of gamma-aminobutyric acid, 1 part of manninotriose and 5 parts of bergamoolide.

The preparation method of the agaric polysaccharide comprises the following steps:

(1) cleaning Auricularia, soaking, homogenizing, extracting with distilled water at 80 deg.C for 30min, boiling for 2 hr, and filtering to obtain primary extractive solution and residue;

(2) adding distilled water into the filter residue, and performing ultrasonic extraction for 60min with ultrasonic power of 80W to obtain secondary extract;

(3) mixing the primary and secondary extractive solutions, concentrating under reduced pressure, and precipitating with ethanol; preparing alcohol precipitate;

(4) dissolving the alcohol precipitate in water, treating the alcohol precipitate by sephadexG-100 gel column, eluting by using ethyl acetate, collecting ethyl acetate eluent, eluting by using distilled water, and collecting distilled water eluent; adding distilled water into ethyl acetate eluate for extraction, mixing the separated water phase with distilled water eluate, and concentrating to obtain Auricularia polysaccharide.

The preparation method of the composition containing pinoresinol diglucoside comprises the following steps:

dissolving gamma-aminobutyric acid in 200 parts of distilled water, adding a mixture of 3:1 propylene glycol and polyethylene glycol laurate (M is 1000) as a solubilizer, wherein the mass concentration of the solubilizer in the distilled water is 1%, and then dispersing pinoresinol diglucoside, syringaresinol, agaric polysaccharide, manninotriose and bergapten in the distilled water to prepare a mixed solution of the composition.

Comparative example 1

The only difference compared to example 1 was the replacement of the auricularia auricula polysaccharide with an equal amount of gamma-aminobutyric acid.

The composition containing pinoresinol diglucoside comprises the following components in parts by weight:

13 parts of pinoresinol diglucoside, 4 parts of syringaresinol, 11 parts of gamma-aminobutyric acid, 5 parts of manninotriose and 11 parts of bergapten.

The preparation method of the composition containing pinoresinol diglucoside comprises the following steps:

dissolving gamma-aminobutyric acid in 200 parts of distilled water, adding a mixture of 2:1 propylene glycol and polyethylene glycol laurate (M is 800) as a solubilizer, wherein the mass concentration of the solubilizer in the distilled water is 2%, and then dispersing pinoresinol diglucoside, syringaresinol, manninotriose and bergamoside lactone in the distilled water to prepare a mixed solution of the composition.

Comparative example 2

The only difference compared to example 1 was the use of the same amount of auricularia auricula polysaccharide instead of gamma-aminobutyric acid.

The composition containing pinoresinol diglucoside comprises the following components in parts by weight:

13 parts of pinoresinol diglucoside, 4 parts of syringaresinol, 11 parts of agaric polysaccharide, 5 parts of manninotriose and 11 parts of bergapten.

The preparation method of the auricularia auricula polysaccharide is the same as that of the example 1.

The preparation method of the composition containing pinoresinol diglucoside comprises the following steps:

taking 200 parts of distilled water, adding a mixture of 2:1 propylene glycol and polyethylene glycol laurate (M is 800) as a solubilizer, wherein the mass concentration of the solubilizer in the distilled water is 2%, and then dispersing pinoresinol diglucoside, syringaresinol, auricularia auricula polysaccharide, manninotriose and bergamoside lactone in the distilled water to prepare a mixed solution of the composition.

Comparative example 3

The only difference compared to example 1 was the use of the same amount of icariin instead of syringaresinol.

The composition containing pinoresinol diglucoside comprises the following components in parts by weight:

13 parts of pinoresinol diglucoside, 4 parts of icariin, 8 parts of agaric polysaccharide, 3 parts of gamma-aminobutyric acid, 5 parts of manninotriose and 11 parts of bergamoolide.

The preparation method of the auricularia auricula polysaccharide is the same as that of the example 1.

The preparation method of the composition containing pinoresinol diglucoside comprises the following steps:

dissolving gamma-aminobutyric acid in 200 parts of distilled water, adding a mixture of 2:1 propylene glycol and polyethylene glycol laurate (M is 800) as a solubilizer, wherein the mass concentration of the solubilizer in the distilled water is 2%, and dispersing pinoresinol diglucoside, icariin, auricularia auricular polysaccharide, manninotriose and bergamoside lactone in the distilled water to prepare a mixed solution of the composition.

Comparative example 4

Compared with example 1, the difference is only the preparation method of the auricularia auricula polysaccharide.

A composition containing pinoresinol diglucoside was prepared as in example 1.

The preparation method of the agaric polysaccharide comprises the following steps:

(1) cleaning Auricularia, soaking, homogenizing, extracting with distilled water at 70 deg.C for 40min, boiling for 1.5 hr, filtering, directly performing ultrasonic treatment for 50min at ultrasonic power of 80W, and filtering to obtain extractive solution and residue;

(2) concentrating the extractive solution under reduced pressure, and precipitating with ethanol; preparing alcohol precipitate;

(4) dissolving the alcohol precipitate in water, treating the alcohol precipitate by sephadexG-100 gel column, eluting by using ethyl acetate, collecting ethyl acetate eluent, eluting by using distilled water, and collecting distilled water eluent; adding distilled water into ethyl acetate eluate for extraction, mixing the separated water phase with distilled water eluate, and concentrating to obtain Auricularia polysaccharide.

The preparation method of the composition containing pinoresinol diglucoside is the same as that of example 1.

Comparative example 5

The only difference compared to example 1 is the solubility promoter.

A composition containing pinoresinol diglucoside was prepared as in example 1.

The preparation method of the agaric polysaccharide comprises the same steps as example 1.

The preparation method of the composition containing pinoresinol diglucoside comprises the following steps:

dissolving gamma-aminobutyric acid in 200 parts of distilled water, adding propylene glycol as a solubilizer, wherein the mass concentration of the solubilizer in the distilled water is 2%, and dispersing pinoresinol diglucoside, syringaresinol, agaric polysaccharide, manninotriose and bergapten in the distilled water to prepare a mixed solution of the composition.

Evaluation of Effect

Rat osteoporosis model: 100 SPF-grade female rats of 3.5 months old are taken, the weight of the rats is 250 +/-20 g, the rats are fed with standard pellet feed at the room temperature of 20-25 ℃, the rats contain 1 percent of calcium and 0.74 percent of phosphorus, the rats are freely fed with food and water, and the experiments are carried out after the rats are adaptively fed for 7 days. Rats were randomly divided into 10 groups, which were a model group, examples 1-3 groups, comparative examples 1-5 groups, and a blank control group.

Wherein, the model group: methylprednisolone 3.5mg/kg/d is injected subcutaneously 1 time a day. Examples 1-3 and comparative examples 1-5 groups: the mixed solution of the composition prepared by the corresponding group of the gavage is 10mg/kg/d (according to the effective content) at the same time of injecting 3.5mg/kg/d of methylprednisolone subcutaneously, and is 1 time a day. Blank control group: 3.5mg/kg/d of physiological saline was injected subcutaneously 1 time a day. Three experiments were performed simultaneously for 8 weeks with 1 dose per week, and the doses were adjusted according to body weight.

After 8 weeks, the bone density of the rats in the model group and the rats in the control group are respectively measured, and the statistical analysis that P is less than 0.05 shows that the osteoporosis model is successfully prepared. After fasting for 1 day, aortic blood was taken, serum was separated, serum osteocalcin (BGP) (kit selected from enzyme-linked organisms) was measured by radioimmunoassay, hip joint bone density (BMD) (EXPERT model manufactured by LUNAR corporation, usa) was measured by using a small animal software dedicated to a dual-energy X-ray bone densitometer, and the test results are shown in table 1.

Table 1 test results data

Note: significant differences (P < 0.05) were indicated between different letters in the same list.

The experimental data show that the model group has significant difference compared with the blank control group, which indicates that the molding is successful; the groups 1 to 3 have significant differences compared with the model group, which shows that the composition provided by the invention can effectively prevent osteoporosis. Secondly, the groups of examples 1 to 3 have significant differences from the groups of comparative examples 1 to 5, which shows that the composition of the present invention and the preparation method thereof have a great influence on the efficacy of treating osteoporosis.

The above detailed description is specific to one possible embodiment of the present invention, and the embodiment is not intended to limit the scope of the present invention, and all equivalent implementations or modifications without departing from the scope of the present invention should be included in the technical scope of the present invention.