CN115068515A - Pharmaceutical composition for treating osteoporosis and application thereof - Google Patents
Pharmaceutical composition for treating osteoporosis and application thereof Download PDFInfo
- Publication number
- CN115068515A CN115068515A CN202210826136.2A CN202210826136A CN115068515A CN 115068515 A CN115068515 A CN 115068515A CN 202210826136 A CN202210826136 A CN 202210826136A CN 115068515 A CN115068515 A CN 115068515A
- Authority
- CN
- China
- Prior art keywords
- quercetin
- ethanol
- extract
- pharmaceutical composition
- astragalus
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Withdrawn
Links
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
- A61K31/35—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom
- A61K31/352—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom condensed with carbocyclic rings, e.g. methantheline
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/48—Fabaceae or Leguminosae (Pea or Legume family); Caesalpiniaceae; Mimosaceae; Papilionaceae
- A61K36/481—Astragalus (milkvetch)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
- A61P19/08—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease
- A61P19/10—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease for osteoporosis
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/331—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using water, e.g. cold water, infusion, tea, steam distillation, decoction
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/39—Complex extraction schemes, e.g. fractionation or repeated extraction steps
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/51—Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/53—Liquid-solid separation, e.g. centrifugation, sedimentation or crystallization
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Animal Behavior & Ethology (AREA)
- Pharmacology & Pharmacy (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Medicinal Chemistry (AREA)
- Physical Education & Sports Medicine (AREA)
- Chemical & Material Sciences (AREA)
- Epidemiology (AREA)
- Orthopedic Medicine & Surgery (AREA)
- Engineering & Computer Science (AREA)
- Rheumatology (AREA)
- Botany (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Medical Informatics (AREA)
- Biotechnology (AREA)
- Alternative & Traditional Medicine (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention provides a pharmaceutical composition for treating osteoporosis and application thereof, wherein the pharmaceutical composition is prepared from quercetin and astragalus extract according to a mass ratio of 1-3: 1-3. The invention combines the quercetin and astragalus extract, has the drug effect which is larger than that of single use, can obviously inhibit LPS (lipopolysaccharide) from promoting osteoclast differentiation, and can obviously reduce the expression of osteoclast-related proteins COX-2, TRAP, TRAF6 and CK. In the medicine compound agent provided by the invention, the dosage of the combined use of the quercetin and the astragalus extract is obviously less than that of the single use.
Description
Technical Field
The invention relates to the technical field of medicines, in particular to a medicinal compound preparation for treating osteoporosis and application thereof.
Background
With the increase of age level, the function of osteoblast is reduced, the bone absorption function of osteoclast is relatively enhanced, so that mineral substances, organic matrixes and the like of bone are gradually reduced, the bone density and the bone quality are gradually reduced, the bone is brittle, and the osteoporosis is easily caused. The pathogenesis of osteoporosis is not clear, and researches show that the increase of PTH (parathyroid hormone) can promote the osteolysis of osteoclast, so that bone calcium enters blood to maintain the blood calcium stability, and osteoporosis is easy to occur. At present, the medicines commonly used for treating osteoporosis mainly comprise calcitonin, calcium vitamin D and the like.
In recent years, it has been found that quercetin can treat castrate osteoporosis rats by promoting osteogenic differentiation. Through the intragastric treatment of 50mg/kg, 100mg/kg and 200mg/kg quercetin in rats with osteoporosis models, the results show that the contents of CTX (collagen type I cross-linked C-terminal peptide), TRACP-5b (tartrate-resistant acid phosphatase 5b), OC (osteocalcin), PINP (procollagen type I N-terminal propeptide) and BALP (bone alkaline phosphatase) in the blood of rats with osteoporosis models are all obviously increased, and Ca, P and BMD (bone density) are obviously reduced; after the stomach is irrigated by the quercetin, when the dosage of the quercetin is lower than 50mg/kg, the improvement effect is not obvious; when the dosage of the quercetin reaches (200mg/kg), the treatment effect basically reaches a normal level.
At present, many reports about the treatment of osteoporosis by quercetin are reported, but in the experimental process of an osteoporosis model mouse, the low dose (less than or equal to 10mg/kg) of quercetin is found to have no obvious effect on the treatment of osteoporosis, while the high dose (more than or equal to 80mg/kg) of quercetin can inhibit the proliferation of osteoblasts and has a certain lethality rate. Therefore, a pharmaceutical composition for treating osteoporosis needs to be researched, so that the osteoporosis is obviously improved while the therapeutic dose of quercetin is reduced, and a better treatment effect is achieved.
Disclosure of Invention
In order to solve the technical problems, the invention aims to provide a pharmaceutical composition for treating osteoporosis and application thereof.
In order to achieve the above object, the technical solution of the present invention is as follows.
A medicine compound agent is prepared by mixing quercetin and astragalus extract according to a mass ratio of 1-3: 1-3;
the astragalus extract is prepared by the following method:
after crushing the astragalus membranaceus, mixing the materials according to a material-liquid ratio of 1 g: adding 8-12 mL of water, performing reflux extraction for 1-3 times, concentrating the extraction solution into an extract, adding ethanol, stirring, standing, centrifuging, adding ethanol into the precipitate, stirring, standing, centrifuging, and combining the two ethanol extraction solutions;
adding mixed solvent into the ethanol extractive solution to remove protein and mucus components to obtain radix astragali extract.
In the invention, the active ingredients of the astragalus extract are flavonoid ingredients and saponin ingredients.
Further, the medicine compound preparation is prepared by mixing quercetin and astragalus extract according to a mass ratio of 1: 1-3.
Further, adding water for reflux extraction for 1-3 h; the specific gravity d of the extract is 1.18-1.22.
And further, adding ethanol to ensure that the volume fraction of the ethanol in the solution is 80-95%, and performing alcohol precipitation.
Further, the mixed solvent is savage reagent.
The invention also provides application of the pharmaceutical composition in preparing a medicament for treating osteoporosis.
Further, the pharmaceutical composition can inhibit the differentiation of the osteoclast promoted by LPS.
Further, the drug complex can down-regulate the expression of osteoclast-associated proteins COX-2, TRAP, TRAF6 and CK.
The invention has the beneficial effects that:
1. the invention uses the quercetin-astragalus extract in combination, the drug effect is larger than that of single use, and the quercetin-astragalus extract can obviously inhibit LPS (lipopolysaccharide) to promote osteoclast differentiation, and can obviously reduce the expression of osteoclast-related proteins COX-2, TRAP, TRAF6 and CK.
2. In the medicine compound agent provided by the invention, the dosage of the combined use of the quercetin and the astragalus extract is obviously less than that of the single use. The medicament compound prepared from the low-dose quercetin and the astragalus extract obviously improves osteoporosis while reducing the treatment dose of the quercetin, and has a better treatment effect.
Drawings
FIG. 1 is a graph showing the results of Western blot detection of osteoclast-associated protein expression in the quercetin-Astragalus membranaceus extract combined treatment test.
FIG. 2 is a graph of the micro-CT detection results of femoral bone specimens from treatment tests of different quercetin dosages and action times. Wherein, A is a bar chart of the activity influence of the single use of different dosages of quercetin on MC3T3-E1 cells at 1d, 3d, 5d and 7d before LPS induction; b is a bar graph of the effect of different doses of quercetin pretreatment on the viability of MC3T3-E1 cells at 1d and 3d before and after LPS induction.
FIG. 3 is a micro-CT detection result chart of femoral bone specimens subjected to single preventive tests by using high-dose quercetin and astragalus extract after LPS induction. Wherein A is a micro-CT scanning image of the cross section of the lower section of the femur; b is a micro-CT reconstructed image of the lower section bone trabecula of the femur; c is a histogram of the bone volume fraction BV/TV statistical analysis.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is described in further detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
All other embodiments, which can be obtained by a person skilled in the art without making any creative effort based on the embodiments in the present invention, belong to the protection scope of the present invention.
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
A pharmaceutical composition for treating osteoporosis is prepared from quercetin and radix astragali extract according to a mass ratio of 1: 2.6 mixing to obtain the product.
The astragalus extract is prepared by the following method:
cleaning, drying and crushing a dried astragalus root raw material, and mixing the raw material with the powder according to a material-liquid ratio of 1 g: adding 10mL of water, and extracting under reflux for 3 times, wherein the extraction time is 2h each time; mixing the three extraction solutions, concentrating the extraction solution into an extract with the specific gravity d of 1.20, adding ethanol, stirring, precipitating with ethanol until the volume fraction of the ethanol in the solution is 95%, standing, centrifuging, adding ethanol into the precipitate, stirring, standing, centrifuging, and mixing the two ethanol extraction solutions; adding savage reagent into the ethanol extractive solution to remove protein and mucus components to obtain radix astragali extract.
Example 2
A pharmaceutical composition for treating osteoporosis is prepared from quercetin and radix astragali extract according to a mass ratio of 1: 3, mixing to obtain the product.
The astragalus extract is prepared by the following method:
cleaning, drying and crushing a dried astragalus root raw material, and mixing the raw material with the water according to a material-liquid ratio of 1 g: adding 8mL of water, and extracting under reflux for 3 times, wherein the extraction time is 1h each time; mixing the three extractive solutions, concentrating the extractive solution to obtain extract with specific gravity d of 1.18, adding ethanol, stirring, precipitating with ethanol until the volume fraction of ethanol in the solution is 90%, standing, centrifuging, adding ethanol into the precipitate, stirring, standing, centrifuging, and mixing the two ethanol extractive solutions; adding savage reagent into the ethanol extractive solution to remove protein and mucus components to obtain radix astragali extract.
Example 3
A pharmaceutical composition for treating osteoporosis is prepared from quercetin and radix astragali extract according to a mass ratio of 1: 3, mixing to obtain the product.
The astragalus extract is prepared by the following method:
cleaning, drying and crushing a dried astragalus root raw material, and mixing the raw material with the liquid material ratio of 1 g: adding 12mL of water, and extracting under reflux for 1 time, wherein the extraction time is 3h each time; concentrating the extract solution to obtain extract with specific gravity d of 1.22, adding ethanol, stirring, precipitating with ethanol until the volume fraction of ethanol in the solution is 80%, standing, centrifuging, adding ethanol into the precipitate, stirring, standing, centrifuging, and mixing the two ethanol extractive solutions; adding savage reagent into the ethanol extractive solution to remove protein and mucus components to obtain radix astragali extract.
Example 4
A pharmaceutical composition for treating osteoporosis is prepared from quercetin and radix astragali extract according to a mass ratio of 1: 2, mixing to obtain the product.
The astragalus extract is prepared by the following method:
cleaning, drying and crushing a dried astragalus root raw material, and mixing the raw material with the powder according to a material-liquid ratio of 1 g: adding 10mL of water, and extracting under reflux for 3 times, wherein the extraction time is 2h each time; mixing the three extractive solutions, concentrating the extractive solution to obtain extract with specific gravity d of 1.20, adding ethanol, stirring, precipitating with ethanol until the volume fraction of ethanol in the solution is 95%, standing, centrifuging, adding ethanol into the precipitate, stirring, standing, centrifuging, and mixing the two ethanol extractive solutions; adding savage reagent into the ethanol extractive solution to remove protein and mucus components to obtain radix astragali extract.
Example 5
A pharmaceutical composition for treating osteoporosis is prepared from quercetin and radix astragali extract according to a mass ratio of 1: 1 are mixed to obtain the product.
The astragalus extract is prepared by the following method:
cleaning, drying and crushing a dried astragalus root raw material, and mixing the raw material with the water according to a material-liquid ratio of 1 g: adding 10mL of water, and extracting under reflux for 3 times, wherein the extraction time is 2h each time; mixing the three extractive solutions, concentrating the extractive solution to obtain extract with specific gravity d of 1.20, adding ethanol, stirring, precipitating with ethanol until the volume fraction of ethanol in the solution is 95%, standing, centrifuging, adding ethanol into the precipitate, stirring, standing, centrifuging, and mixing the two ethanol extractive solutions; adding savage reagent into the ethanol extractive solution to remove protein and mucus components to obtain radix astragali extract.
Example 6
A pharmaceutical composition for treating osteoporosis is prepared from quercetin and radix astragali extract according to a mass ratio of 3: 1 are mixed to obtain the product.
The astragalus extract is prepared by the following method:
cleaning, drying and crushing a dried astragalus root raw material, and mixing the raw material with the powder according to a material-liquid ratio of 1 g: adding 10mL of water, and extracting under reflux for 3 times, wherein the extraction time is 2h each time; mixing the three extractive solutions, concentrating the extractive solution to obtain extract with specific gravity d of 1.20, adding ethanol, stirring, precipitating with ethanol until the volume fraction of ethanol in the solution is 95%, standing, centrifuging, adding ethanol into the precipitate, stirring, standing, centrifuging, and mixing the two ethanol extractive solutions; adding savage reagent into the ethanol extractive solution to remove protein and mucus components to obtain radix astragali extract.
The effects of the pharmaceutical compositions prepared in examples 1 to 6 are substantially the same, and the pharmaceutical composition of example 1 is tested in a mouse model for osteoporosis to illustrate the mechanism of action of the pharmaceutical composition on osteoporosis.
1. Test animal and reagent
1.1 test animals
Female C57BL/6 mice (8 weeks, 20-25 g in physical quality) were bred under standard conditions, provided by the academy of medical sciences of Henan province (Zheng, China). All experimental procedures were approved, according to the care and use of the experimental animals. All efforts have been made to reduce the pain and pain of the animals tested.
1.2, reagents
The LPS injection solution was prepared to be 100ng/mL with sterile water for injection.
2. LPS mouse model construction
Mice were induced to a mouse osteoporosis model by intraperitoneal injection of LPS injection for 2 weeks (5mg/kg/d,3 times/week), and the mental state, mobility, etc. of the mice were observed.
3. Grouping of test animals
3.1 Quercetin-Astragalus membranaceus extract combination treatment test
27 mice were randomly assigned to nine groups: a control group (control), an LPS model group (LPS), a quercetin LOW dose group (LOW-Q), a quercetin HIGH dose group (HIGH-Q), an astragalus extract LOW dose group (LOW-A), an astragalus extract HIGH dose group (HIGH-A), a quercetin and astragalus extract LOW dose group (LOW-Q/A), a quercetin and astragalus extract medium dose group (MID-Q/A) and a quercetin and astragalus extract HIGH dose group (HIGH-Q/A); three in each group.
a) Control group (control): the same amount of physiological saline was intraperitoneally injected, and LPS was not intraperitoneally injected.
b) LPS model group (LPS): the same amount of normal saline was intraperitoneally injected after LPS induction.
c) Quercetin LOW dose group (LOW-Q): 5mg/kg quercetin was intraperitoneally injected after LPS induction.
d) Quercetin HIGH dose group (HIGH-Q) 20mg/kg quercetin was intraperitoneally injected after LPS induction.
e) Low dose group of radix astragali extract (LOW-A) is prepared by injecting radix astragali extract 13mg/kg into abdominal cavity after LPS induction.
f) HIGH dose group of radix astragali extract (HIGH-A) is prepared by intraperitoneal injection of 50mg/kg radix astragali extract after LPS induction.
g) Quercetin and radix astragali extract Low dose group (LOW-Q/A) is prepared by intraperitoneal injection of 1.5mg/kg quercetin and 4mg/kg radix astragali extract after LPS induction.
h) Quercetin in combination with radix astragali extract middle dose group (MID-Q/A) is administered by intraperitoneal injection of 3mg/kg quercetin +8mg/kg radix astragali extract after LPS induction.
j) Quercetin in combination with radix astragali extract HIGH dose group (HIGH-Q/A) LPS induced intraperitoneal injection of 6mg/kg quercetin +16mg/kg radix astragali extract.
3.2 treatment trials with different Quercetin doses and duration of action
39 mice were randomly assigned to nine groups: a blank control group, a quercetin positive control group, an LPS model group, a quercetin treatment group, a quercetin treatment group, a control group, a quercetin treatment group, a control, a quercetin treatment group, a control, a LPS model, a control; three in each group.
Blank control group: the same amount of physiological saline was intraperitoneally injected, and LPS was not intraperitoneally injected.
Quercetin positive control group: 5mg/kg quercetin was intraperitoneally injected, and LPS was not intraperitoneally injected.
Two groups of quercetin positive controls: 20mg/kg quercetin was intraperitoneally injected, and LPS was not intraperitoneally injected.
Quercetin positive control three groups: 50mg/kg quercetin was intraperitoneally injected, and LPS was not intraperitoneally injected.
LPS model group: the same amount of normal saline was intraperitoneally injected after LPS induction.
Quercetin treatment one group: 5mg/kg quercetin was intraperitoneally injected after LPS induction.
Quercetin treated two groups: after LPS induction, 10mg/kg quercetin was intraperitoneally injected.
Quercetin treated three groups: after LPS induction, 20mg/kg quercetin was intraperitoneally injected.
Quercetin treated four groups: after LPS induction, 40mg/kg quercetin was intraperitoneally injected.
Quercetin treatment five groups: after LPS induction, 60mg/kg quercetin was intraperitoneally injected.
Quercetin treatment six groups: after LPS induction, 80mg/kg quercetin was intraperitoneally injected.
Quercetin treatment of seven groups: after LPS induction, 100mg/kg quercetin was intraperitoneally injected.
Quercetin treatment of eight groups: after LPS induction, 150mg/kg quercetin was intraperitoneally injected.
4. Test method
4.1 Quercetin-Astragalus membranaceus extract combined treatment test
Performing a quercetin-astragalus extract combined treatment test according to a grouping method of 3.1, for a test group b) to a test group j), firstly inducing RAW264.7 cells for 2d by using 100ng/ml LPS, then performing combined treatment on the quercetin-astragalus extract for 1d, and detecting osteoclast-related protein expression by Western blot; the results are shown in FIG. 1.
4.2 treatment trials with different Quercetin doses and durations of action
Performing treatment tests of different quercetin dosages and action time according to the grouping method of 3.2, and performing separate pretreatment on quercetin (0, 5, 20, 50mg/kg) with different dosages for 1 week; mice were then given LPS (lipopolysaccharide) (5mg/kg/d,3 times per week) and were injected intraperitoneally with LPS for 2 weeks in combination with different doses of quercetin (0, 5, 10, 20, 40m, 60, 80, 100, 150 g/kg). All test mice were sacrificed by cervical dislocation 1 day after completion of the experiment. Taking the right femur of the mouse for micro-CT detection; the results are shown in FIG. 2.
The micro-CT detection uses a Bruke rMicroCT Skyscan 1272 system, the equidirectional three-dimensional pixels adopt 10.0 mu m, and the scanning object is the whole femur; separating and fully exposing the thighbone of the mouse before scanning, removing muscle tissues and soft tissues, and fixing for 24 hours by using paraformaldehyde; the voltage of a scanning X-ray tube is 60kV, the X-ray intensity is 166 muA, and the exposure time is 1700 ms; analyzing the trabecula femoris at the far end by adopting a mode of being 0.8mm from the near end of the central point of the epiphyseal end to 3mm from the far end; cortical bone analysis (2D analysis) using 0.5mm proximal to the epiphyseal centre point to 4.5mm distal; the trabecular bone and cortical bone analysis both adopt a threshold value of 86-255; scanning the whole body of the mouse by adopting an equidirectional voxel 148 mu m; the reconstruction of the image uses software Nrecon (ver.1.6.10); the 3D image is generated by the 2D image according to the gray value by using a distance transformation algorithm and software CTvox (Ver.3.0.0); the 2D and 3D images were analyzed by CT Analyzer (Ver.1.15.4.0).
4.3, preventive test alone
Performing individual preventive tests by using LPS in combination with high-dose quercetin and LPS in combination with high-dose radix astragali extract according to the grouping method of 3.1, and performing individual preventive intraperitoneal injection for test group a), test group b), test group d) and test group f) with low-dose quercetin and radix astragali extract for 1 week; mice were administered LPS (5mg/kg/d,3 times/week), quercetin (20mg/kg/d, 3 times/week), LPS in combination with the extract of Astragalus (20mg/kg/d, 3 times/week) in combination with LPS for 2 weeks. Killing the mouse, and carrying out micro-CT scanning on a femoral bone specimen; the results are shown in FIG. 3.
5. Test results and discussion
5.1, as can be seen from FIG. 1, the combined treatment of quercetin-Astragalus extract can significantly inhibit the differentiation of LPS-promoted osteoclasts, which is superior to that of the single treatment group. Western blot detection finds that the low-dose quercetin-astragalus extract group obviously reduces the expression of osteoclast-associated proteins COX-2, TRAP, TRAF6 and CK, and is superior to a single treatment group.
5.2, as can be seen from fig. 2, the micro-CT scanning of the femur bone specimen shows that the low dose quercetin and the astragalus extract can not reverse inflammatory bone loss caused by LPS when being used alone.
5.3, as can be seen from FIG. 3, the micro-CT scan of the femoral bone specimen shows that the low dose quercetin and the astragalus extract can be used for singly and prophylactically treating the LPS inflammation model mouse for 2 weeks, and the reduction of the femoral bone mass of the mouse caused by LPS can not be reduced.
6. Conclusion of the experiment
The invention uses the quercetin-astragalus extract in combination, the drug effect is larger than that of single use, and the quercetin-astragalus extract can obviously inhibit LPS (lipopolysaccharide) to promote osteoclast differentiation, and can obviously reduce the expression of osteoclast-related proteins COX-2, TRAP, TRAF6 and CK. The medicament compound prepared from low-dose quercetin and astragalus extract can obviously improve osteoporosis while reducing the treatment dose of quercetin, and has a good treatment effect.
The present invention is not limited to the above preferred embodiments, and any modifications, equivalent substitutions and improvements made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (8)
1. The pharmaceutical composition for treating osteoporosis is characterized by comprising quercetin and astragalus extract according to a mass ratio of 1-3: 1-3;
the astragalus extract is prepared by the following method:
after crushing the astragalus membranaceus, mixing the materials according to a material-liquid ratio of 1 g: adding 8-12 mL of water, performing reflux extraction for 1-3 times, concentrating the extraction solution into an extract, adding ethanol, stirring, standing, centrifuging, adding ethanol into the precipitate, stirring, standing, centrifuging, and combining the two ethanol extraction solutions;
adding mixed solvent into the ethanol extractive solution to remove protein and mucus components to obtain radix astragali extract.
2. The pharmaceutical composition for treating osteoporosis of claim 1, wherein the pharmaceutical composition is prepared from quercetin and astragalus extract according to a mass ratio of 1: 1-3.
3. The pharmaceutical composition for treating osteoporosis of claim 1, wherein the time of water reflux extraction is 1-3 h; the specific gravity d of the extract is 1.18-1.22.
4. The pharmaceutical composition for treating osteoporosis of claim 1, wherein ethanol is added to the solution so that the volume fraction of ethanol in the solution is 80-95% for ethanol precipitation.
5. The pharmaceutical composition for treating osteoporosis of claim 1, wherein the mixed solvent is savage reagent.
6. Use of the pharmaceutical combination according to claim 1 for the preparation of a medicament for the treatment of osteoporosis.
7. The use according to claim 6, wherein the pharmaceutical combination is capable of inhibiting LPS osteoclast differentiation.
8. The use according to claim 6, wherein the pharmaceutical combination is capable of down-regulating the expression of the osteoclast-associated proteins COX-2, TRAP, TRAF6 and CK.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210826136.2A CN115068515A (en) | 2022-07-14 | 2022-07-14 | Pharmaceutical composition for treating osteoporosis and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210826136.2A CN115068515A (en) | 2022-07-14 | 2022-07-14 | Pharmaceutical composition for treating osteoporosis and application thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN115068515A true CN115068515A (en) | 2022-09-20 |
Family
ID=83259528
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210826136.2A Withdrawn CN115068515A (en) | 2022-07-14 | 2022-07-14 | Pharmaceutical composition for treating osteoporosis and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN115068515A (en) |
-
2022
- 2022-07-14 CN CN202210826136.2A patent/CN115068515A/en not_active Withdrawn
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102172356B (en) | Use of taurochenodeoxycholic acid in prevention and treatment of osteoporosis | |
| KR100679778B1 (en) | FRACTURE HEALING USING PTHrP ANALOGS | |
| KR100345752B1 (en) | Pharmacolocial effect and extracting method for chronic osteoporosis and degenerative bone marrow diseases and rhematoid arthritis treatment by constituent drugs of oriental medicine | |
| Weng et al. | Combined treatment with Cinnamaldehyde and β-TCP had an additive effect on bone formation and angiogenesis in critical size calvarial defect in ovariectomized rats | |
| CN103798595A (en) | Healthcare food for increasing bone mineral density and preparation method thereof | |
| Jiang et al. | The effects of hypoxia-inducible factor (HIF)-1α protein on bone regeneration during distraction osteogenesis: an animal study | |
| CN107349333B (en) | Composition with effect of increasing bone mineral density and preparation method and application thereof | |
| CN113713067B (en) | Traditional Chinese medicine composition for preventing and treating sarcopenia osteoporosis and preparation method thereof | |
| CN109453188B (en) | Application of sandworm polysaccharide in preparation of medicine for preventing and treating osteoporosis | |
| Liu et al. | Protection effect of curcumin for macrophage-involved polyethylene wear particle-induced inflammatory osteolysis by increasing the cholesterol efflux | |
| CN115068515A (en) | Pharmaceutical composition for treating osteoporosis and application thereof | |
| CN101167754A (en) | Application of sea bone peptide in preparing medicine, health-care food or food for preventing osteoporosis | |
| CN113116897A (en) | Application of magnoflorine in preparation of bone regulation drug synergist and drug composition containing magnoflorine | |
| CN1781510A (en) | Use of sea-buckthorn fruit oil and/or fruit slag oil in preparing medicine for treating osteoporosis | |
| CN109320600B (en) | Novel PTH mimic peptide based on protein domain reconstruction and application thereof | |
| CN116212007B (en) | Bone-knitting and tendon-softening traditional Chinese medicine composition for treating metaphase fracture and preparation method thereof | |
| CN103316143B (en) | Chinese medicine composition for inhibiting rheumatoid arthritis inflammation and reducing bone mass loss and application thereof | |
| CN107625849B (en) | Application of Wuzi Yanzong pills | |
| CN109223771B (en) | Application of brucine A in preparing medicine for preventing and treating osteoporosis | |
| CN104523685A (en) | Use of deferasirox in preparation of medicine for treating postmenopausal osteoporosis diseases | |
| CN110711201A (en) | Application of punicalagin in preparing medicine for preventing and treating postmenopausal osteoporosis | |
| CN110898089A (en) | Traditional Chinese medicine extract composition for treating osteoporosis | |
| Zhang et al. | Prevention and treatment of osteoporosis caused by oestrogen shortage in rats. | |
| CN117137924A (en) | Application of N-acetyl-D-mannosamine in preparation of food and medicine for promoting bone growth | |
| WO2019061862A1 (en) | Shell nacre matrix protein, preparation method therefor, and use thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WW01 | Invention patent application withdrawn after publication |
Application publication date: 20220920 |
|
| WW01 | Invention patent application withdrawn after publication |