CN109549943B - Pharmaceutical composition for promoting periodontal bone repair - Google Patents
Pharmaceutical composition for promoting periodontal bone repair Download PDFInfo
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7028—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages
- A61K31/7034—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/36—Polysaccharides; Derivatives thereof, e.g. gums, starch, alginate, dextrin, hyaluronic acid, chitosan, inulin, agar or pectin
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- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0053—Mouth and digestive tract, i.e. intraoral and peroral administration
- A61K9/006—Oral mucosa, e.g. mucoadhesive forms, sublingual droplets; Buccal patches or films; Buccal sprays
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- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/02—Stomatological preparations, e.g. drugs for caries, aphtae, periodontitis
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/70—Web, sheet or filament bases ; Films; Fibres of the matrix type containing drug
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Abstract
The invention belongs to the field of medicines, and particularly relates to a pharmaceutical composition for promoting periodontal bone repair. The pharmaceutical composition takes the phenylethanoid glycosides compound as an active ingredient, can promote the proliferation of osteoblasts and promote the bone repair of periodontal diseases, and has more excellent corresponding effect after being used with chitosan.
Description
Technical Field
The invention belongs to the field of medicines, and particularly relates to a pharmaceutical composition for promoting periodontal bone repair.
Background
Periodontal disease refers to a chronic inflammatory disease that occurs in the periodontal tissues (including gingiva, periodontal ligament, alveolar bone, cementum, etc.). Periodontal disease is not only one of the clinical causes of oral cavity problems such as periodontal abscess, gingival bleeding, gingival atrophy, halitosis, etc., but also causes the degradation of periodontal tissues, the absorption of alveolar bone, and finally leads to the loosening and even the falling of teeth, wherein the destruction of periodontal tissues and the absorption of alveolar bone are typical clinical pathological changes of periodontal disease.
For periodontal disease, at present, clinically, methods such as anti-inflammation, tooth restoration, root canal treatment and the like are adopted according to pathogenic factors, the development of periodontal disease can be relieved to a certain extent, and alveolar bone absorption is delayed, so that tooth loosening and falling are avoided to a certain extent.
The phenylethanoid glycosides compounds are formed by connecting acrylic acid and phenylethanol with beta-glucopyranose through ester bonds and glycosidic bonds respectively, and can be specifically divided into:
(1) phenylethanoid glycosides in which the C-3' position of glucose is substituted by α -L-rhamnose, such as:
(2) phenylethanoid glycosides, unsubstituted or substituted at the C-3' position with a sugar other than α -L-rhamnose, such as:
(3) phenylethanoid glycosides having a substituent at the C-7 position of the phenylethanol group, such as:
(4) phenylethanoid glycosides containing 1, 4-dioxygen six-membered rings, such as:
(5) phenylethanoid glycosides containing an iridoid group, such as:
(6) phenylethanoid glycosides with glucose attached to the phenethyl-4 hydroxyl group, such as:
(7) the core sugar is phenylethanoid glycoside of glycosyl except glucose, such as:
the phenylethanoid glycosides have been reported to have multiple biological activities, including anti-tumor, anti-virus, anti-bacterial, anti-inflammatory, liver-protecting, anti-oxidation, immune regulation, neuroprotection, etc., but the use of the phenylethanoid glycosides for periodontal disease, especially for promoting bone repair of periodontal disease, has not been reported.
Disclosure of Invention
The invention aims to provide a pharmaceutical composition for promoting periodontal bone repair.
In order to realize the purpose, the invention adopts the following technical scheme:
a pharmaceutical composition for promoting periodontal bone repair contains phenylethanoid glycosides as active ingredients.
Optionally, the phenylethanoid glycoside compound has the following structure:
wherein R1 and R2 are independently selected from hydrogen or glycosyl; r3 and R5 are independently selected from hydrogen or hydroxyl; r4 is selected from hydrogen, acetyl or glycosyl.
Optionally, the glycosyl is selected from: glucopyranosyl, alpha-L-rhamnosyl and galactosyl.
Optionally, the phenylethanoid glycosides compounds are selected from: one or more of herba cistanches Deserticolae A, herba cistanches Deserticolae B1, herba cistanches Deserticolae B2, herba cistanches Deserticolae C1, and herba cistanches Deserticolae C2.
Further, the pharmaceutical composition comprises the phenylethanoid glycoside compound and a pharmaceutically acceptable carrier.
Further, the pharmaceutical composition comprises the phenylethanoid glycoside compound, chitosan and a pharmaceutically acceptable carrier.
Optionally, the chitosan is selected from: carboxymethyl chitosan.
Optionally, the weight ratio of the phenylethanoid glycosides compound to the chitosan in the pharmaceutical composition is 1-10: 3-20.
Further, the weight ratio of the phenylethanoid glycosides compound to the chitosan is 3-8: 6-15;
furthermore, the weight ratio of the phenylethanoid glycosides compound to the chitosan is 6: 11.
optionally, the content of the phenylethanoid glycosides compounds and the chitosan in the pharmaceutical composition accounts for 5-30% of the total weight of the pharmaceutical composition.
Furthermore, the content of the phenylethanoid glycosides compounds and the chitosan accounts for 6-20% of the total weight of the pharmaceutical composition;
furthermore, the content of the phenylethanoid glycosides compounds and the chitosan accounts for 8-15% of the total weight of the pharmaceutical composition;
furthermore, the content of the phenylethanoid glycosides compounds and the chitosan accounts for 10% of the total weight of the pharmaceutical composition.
Optionally, the pharmaceutical composition is a gel.
In another aspect, the preparation method of the pharmaceutical composition for promoting periodontal bone repair comprises the following steps:
(1) crushing: respectively pulverizing phenylethanoid glycosides compounds and chitosan, sieving with 30-60 mesh sieve, and mixing to obtain mixed powder;
(2) preparation: adding pharmaceutically acceptable adjuvants into the mixed powder, and making into gel.
Compared with the prior art, the invention has the beneficial effects that: the invention provides a pharmaceutical composition for promoting bone repair of periodontal diseases, which is prepared by applying phenylethanoid glycosides compounds to drugs for treating and preventing periodontal diseases for the first time, inhibiting periodontal inflammatory alveolar bone resorption, promoting osteoblast differentiation, promoting bone repair related to periodontal diseases, and has excellent effect when used together with chitosan. The pharmaceutical composition for promoting periodontal bone repair does not need to add preservatives, is safe and free of side effects, the gel is applied through oral mucosa, active ingredients are slowly released, the action time is long, the active ingredients are not influenced by liver first-pass effect, no gastrointestinal irritation exists, the application is convenient, and the compliance of patients is high.
Detailed Description
The present invention is further illustrated by the following specific examples.
Example 1A gel for promoting periodontal bone repair
The traditional Chinese medicine composition is prepared from 26 parts of cistanche tubulosa glucoside B, 11 parts of carboxymethyl chitosan, 50 parts of sterilized water and 1 part of flavoring agent by the following method:
(1) crushing: respectively pulverizing herba cistanches Deserticolae B2 and carboxymethyl chitosan, sieving with 40 mesh sieve, and mixing to obtain mixed powder;
(2) preparation: adding a flavoring agent and sterilized water into the mixed powder, uniformly stirring, fully swelling, and packaging to obtain the gel for promoting periodontal bone repair.
Example 2-6A gel for promoting periodontal bone repair
| Components | Example 2 | Example 3 | Example 4 | Example 5 | Example 6 |
| Cistanche tubulosa glucoside B2 | 8.5 | 2 | |||
| Cistanche tubulosa Roxb | 6 | ||||
| Cistanche tubulosa glucoside B1 | 6 | ||||
| Cistanche tubulosa C1 | 6 | ||||
| Cistanche tubulosa C2 | |||||
| Carboxymethyl chitosan | 8.5 | 15 | 11 | 11 | 11 |
| Flavouring agent | 1 | 1 | 1 | 1 | 1 |
| Sterilized water | 50 | 50 | 50 | 50 | 50 |
The gels for promoting periodontal bone repair described in examples 2-6 were prepared as described in example 1.
Effect example 1 Effect of Phenylethanoid glycosides on proliferation of rat osteoblasts
1.1 Experimental drugs:
herba cistanches Deserticolae A, herba cistanches Deserticolae B1, herba cistanches Deserticolae B2, herba cistanches Deserticolae C1, and herba cistanches Deserticolae C2, and preparing into 0.25g/L liquid with sterile PBS solution for use.
1.2 Experimental methods
(1) Taking calvaria bone: taking neonatal SD rat calvaria bone under aseptic condition, removing connective tissue and blood vessel adhered on the rat calvaria bone, washing with sterile PBS solution for 5 times, and cutting with sterile surgical scissors;
(2) removing fibroblasts: adding 3g/L trypsin into the minced rat calvaria bone, digesting for 15min at 37 ℃, removing digestive juice by suction, and washing for 3 times by using sterile PBS;
(3) digestion of osteoblasts: adding the mixture in a volume ratio of 1: 1, 1.5g/L collagenase type I and 1g/L hyaluronidase, digesting at 37 deg.C for 20min, sucking out the digestive juice for standby, digesting fully for 5 times, collecting the digestive juice of 3-5 times, centrifuging, discarding the supernatant, washing with serum-free DMEM medium, centrifuging, discarding the supernatant, adding 10% fetal calf serum-containing DMEM medium, and uniformly blowing to obtain the product3×105The cells obtained were inoculated in a cell culture flask at a concentration of 5% CO at 37 ℃2In the incubator, the medium was changed 1 time every 48 hours, and after the cells were fused, they were digested with 2.5g/L trypsin and then passaged. 3 rd generation cells were selected at 3X 104Inoculating the cells into a 96-well plate at a density of/mL, adding a DMEM culture medium containing 10% fetal calf serum, wherein each well is 100 mu L, dividing the inoculated cells into a blank group and an experiment 1-5 group, repeating the 3 groups of the blank group, sucking off a supernatant after inoculating for 24 hours, adding a DMEM culture medium containing 10% fetal calf serum again, each well is 90 mu L, adding test medicaments which are blown and beaten uniformly according to the groups, each well is 10 mu L, wherein the blank group is added with a sterile PBS solution, continuously culturing for 48 hours, then adding a 5g/L MTT solution, each well is 20 mu L, continuously culturing for 4 hours, sucking off the culture solution, adding DMSO, each well is 150 mu L, shaking for 5 minutes, detecting the absorbance at 570nm by using an enzyme labeling instrument, and calculating the promotion rate of each test medicament on osteoblast proliferation, wherein the specific experimental results are shown in Table 1:
the promotion rate (%) - (experimental absorbance-blank absorbance)/blank absorbance × 100%.
1.3 results of the experiment
Data analysis was performed using the multi-factor analysis of variance module of statistical software SPSS, where P <0.05 indicates that the difference is statistically significant.
TABLE 1 Effect of Phenylethanoid glycosides on rat osteoblast proliferation
| Group of | Absorbance of the solution | Acceleration Rate (%) |
| Blank group | 0.596±0.047 | |
| Cistanche tubulosa Roxb | 0.675±0.068* | 13.26％ |
| Cistanche tubulosa glucoside B1 | 0.742±0.054* | 24.50％ |
| Cistanche tubulosa glucoside B2 | 0.839±0.076** | 40.77％ |
| Cistanche tubulosa C1 | 0.663±0.051* | 11.24％ |
| Cistanche tubulosa C2 | 0.724±0.065* | 21.48％ |
P <0.05, P <0.01 compared to blank group.
The experimental results in table 1 show that the cistanche tubulosa glycoside A, the cistanche tubulosa glycoside B1, the cistanche tubulosa glycoside B2, the cistanche tubulosa glycoside C1 and the cistanche tubulosa glycoside C2 all show the effect of promoting the proliferation of the osteoblasts of the rat, wherein the effect of the cistanche tubulosa glycoside B1, the effect of the cistanche tubulosa glycoside B2 and the effect of the cistanche tubulosa glycoside C2 are more excellent, the effect of the cistanche tubulosa glycoside B2 is most excellent, the promotion rate relative to the blank group exceeds 40%, and the phenylethanoid glycoside compound, especially the cistanche tubulosa glycoside B2 has the remarkable effect of promoting the proliferation of the osteoblasts of the rat.
Effect example 2: effect of combination of cistanche tubulosa B2 and carboxymethyl chitosan on proliferation of rat osteoblasts
2.1 Experimental drugs
The following experimental drugs were prepared into 0.25g/L liquid using sterile PBS solution for further use.
TABLE 2 Experimental drugs
| Group of | Experimental drugs |
| Medicine 1 | Cistanche tubulosa glucoside B2 |
| Medicine 2 | Carboxymethyl chitosan |
| Medicine 3 | Herba cistanches Deserticolae B2 and carboxymethyl chitosan 6:1 |
| Medicine 4 | Herba cistanches Deserticolae B2 and carboxymethyl chitosan 6: 2 |
| Medicine 5 | Herba cistanches Deserticolae B2 and carboxymethyl chitosan 6: 4 |
| Medicine 6 | Herba cistanches Deserticolae B2 and carboxymethyl chitosan 6: 6 |
| Medicine 7 | Herba cistanches Deserticolae B2 and carboxymethyl chitosan 6: 8 |
| Medicine 8 | Herba cistanches Deserticolae B2 and carboxymethyl chitosan 6: 9 |
| Medicine 9 | Herba cistanches Deserticolae B2 and carboxymethyl chitosan 6: 10 |
| Medicament 10 | Herba cistanches Deserticolae B2 and carboxymethyl chitosan 6:11 |
| Medicament 11 | Herba cistanches Deserticolae B2 and carboxymethyl chitosan 6: 12 |
| Medicament 12 | Herba cistanches Deserticolae B2 and carboxymethyl chitosan 6: 13 |
| Medicine 13 | Herba cistanches Deserticolae B2 and carboxymethyl chitosan 6: 16 |
| Medicament 14 | The ratio of the cistanche tubulosa glucoside B2 to the carboxymethyl chitosan is 6: 20 |
2.2 Experimental methods
The rate of promotion of rat osteoblasts by each test drug was determined according to the method described in example 1, and the results are shown in Table 3.
2.3 results of the experiment
Data analysis was performed using the multi-factor analysis of variance module of statistical software SPSS, where P <0.05 indicates that the difference is statistically significant.
The experimental results in Table 3 show that carboxymethyl chitosan alone does not have the effect of promoting the proliferation of rat osteoblasts, and the ratio of the combination of cistanche tubulosa B2 and carboxymethyl chitosan has a significant effect on the effect of promoting the proliferation of rat osteoblasts, and can be roughly divided into three stages: the first stage is as follows: when the content of the cistanche tubulosa B2 is more than that of the carboxymethyl chitosan, the effect of the composition for promoting the proliferation of the rat osteoblasts is gradually reduced along with the increase of the content of the carboxymethyl chitosan; and a second stage: when the content of the cistanche tubulosa deserticola B2 is less than that of the carboxymethyl chitosan and the ratio of the cistanche tubulosa deserticola B2 to the carboxymethyl chitosan is more than or equal to 6:11, the effect of the composition for promoting the proliferation of the rat osteoblasts is gradually enhanced, when the ratio of the cistanche tubulosa deserticola B2 to the carboxymethyl chitosan is 6:11, the composition has the obvious effect of promoting the proliferation of the rat osteoblasts and the effect is superior to the effect of the cistanche tubulosa deserticola B2 alone, and the effect of promoting the proliferation of the rat osteoblasts is enhanced when the ratio of the cistanche tubulosa deserticola B2 to the carboxymethyl chitosan is 6: 11; and a third stage: when the ratio of the cistanche tubulosa deserticola B2 to the carboxymethyl chitosan is less than 6:11, the effect of the composition on promoting the proliferation of the rat osteoblasts is gradually reduced, and when the ratio of the cistanche tubulosa deserticola B2 to the carboxymethyl chitosan is 6: 20, the composition has almost no effect on promoting the proliferation of the rat osteoblasts.
TABLE 3 Effect of combinations of cistanche tubulosa B2 with carboxymethyl chitosan on proliferation of rat osteoblasts
| Group of | Test drugs | Absorbance of the solution | Acceleration Rate (%) |
| Blank group | 0.531±0.044 | ||
| Medicine 1 | Cistanche tubulosa glucoside B2 | 0.725±0.057** | 36.53％ |
| Medicine 2 | Carboxymethyl chitosan | 0.486±0.061 | -8.47％ |
| Medicine 3 | Herba cistanches Deserticolae B2 and carboxymethyl chitosan 6:1 | 0.659±0.068* | 24.11％ |
| Medicine 4 | Herba cistanches Deserticolae B2 and carboxymethyl chitosan 6: 2 | 0.592±0.053 | 11.49％ |
| Medicine 5 | Herba cistanches Deserticolae B2 and carboxymethyl chitosan 6: 4 | 0.617±0.066* | 16.20％ |
| Medicine 6 | Herba cistanches Deserticolae B2 and carboxymethyl chitosan 6: 6 | 0.580±0.076 | 9.23％ |
| Medicine 7 | Herba cistanches Deserticolae B2 and carboxymethyl chitosan 6: 8 | 0.629±0.054* | 18.46％ |
| Medicine 8 | Herba cistanches Deserticolae B2 and carboxymethyl chitosan 6: 9 | 0.658±0.069* | 23.92％ |
| Medicine 9 | Herba cistanches Deserticolae B2 and carboxymethyl chitosan 6: 10 | 0.714±0.081** | 34.46％ |
| Medicament 10 | Herba cistanches Deserticolae B2 and carboxymethyl chitosan 6:11 | 0.787±0.079** | 48.21％ |
| Medicament 11 | Herba cistanches Deserticolae B2 and carboxymethyl chitosan 6: 12 | 0.705±0.094** | 32.77％ |
| Medicament 12 | Herba cistanches Deserticolae B2 and carboxymethyl chitosan 6: 13 | 0.673±0.061* | 26.74％ |
| Medicine 13 | Herba cistanches Deserticolae B2 and carboxymethyl chitosan 6: 16 | 0.624±0.048* | 17.51％ |
| Medicament 14 | The ratio of the cistanche tubulosa glucoside B2 to the carboxymethyl chitosan is 6: 20 | 0.539±0.053 | 1.51％ |
P <0.05, P <0.01 compared to blank group.
The foregoing describes preferred embodiments of the present invention, but is not intended to limit the invention thereto. Modifications and variations of the embodiments disclosed herein may be made by those skilled in the art without departing from the scope and spirit of the invention.
Claims (2)
1. The pharmaceutical composition for promoting periodontal bone repair is characterized by comprising a phenylethanoid glycosides compound, chitosan and a pharmaceutically acceptable carrier, wherein the phenylethanoid glycosides compound is cistanche tubulosa glycoside B2, and the chitosan is selected from the following group: the weight ratio of the carboxymethyl chitosan to the cistanche tubulosa B2 is 6:11, and the pharmaceutical composition for promoting periodontal bone repair is prepared according to the following steps:
(1) crushing: respectively pulverizing herba cistanches Deserticolae B2 and carboxymethyl chitosan, sieving with 30-60 mesh sieve, and mixing to obtain mixed powder;
(2) preparation: adding pharmaceutically acceptable adjuvants into the mixed powder, and making into gel.
2. The pharmaceutical composition for promoting periodontal bone repair according to claim 1, wherein the contents of the phenylethanoid glycosides and the chitosan in the pharmaceutical composition are 5-30% of the total weight of the pharmaceutical composition.
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| CN103622980A (en) * | 2013-12-12 | 2014-03-12 | 宁夏医科大学 | Application of cistanche phenylethanoid glycoside compound to preparation of drugs for treating osteoporosis and drug composition containing cistanche phenylethanoid glycoside compound |
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| CN1268341C (en) * | 2003-03-04 | 2006-08-09 | 杏辉天力（杭州）药业有限公司 | Tubiflorous desert cistanche prepn containing phenethyl alcohol glycoside and its prepn process and use |
| CN102643330B (en) * | 2011-07-09 | 2018-10-19 | 广东医科大学 | A kind of synthetic method and its medical application of bone target medicine Aspartate hexapeptide Danshensu |
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| 羧甲基壳聚糖治疗慢性牙周炎骨缺损的疗效观察;张永军 等;《承德医学院学报》;20141231;第487-488页 * |
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