Pharmaceutical composition for preventing and treating gingival atrophy and preparation method and application thereof

Abstract

The invention provides a pharmaceutical composition for preventing and treating gingival atrophy and a preparation method and application thereof, and belongs to the technical field of biological medicine. The composition contains finless eel peptide, skipjack elastin peptide, quinoa rice extract, gleditsia sinensis rice extract, calendula extract, chamomile extract, chitosan, peach gum, hydroxyapatite and the like, and the finless eel peptide, the skipjack elastin peptide, the quinoa rice extract and the gleditsia sinensis rice extract are specific enzymatic hydrolysates. The monopterus albus peptide, the skipjack elastin peptide, the quinoa rice extract and the gleditsia sinensis rice extract jointly promote repair and regeneration of gingival tissues. The calendula extract, the chamomile extract and the chitosan have anti-inflammatory and antibacterial effects, so that gingival inflammation is relieved, and bacterial infection is prevented. And the chitosan and the calendula extract can accelerate the repair of damaged gingival tissues. The monopterus albus peptide, the skipjack elastin peptide and the hydroxyapatite are matched for use to jointly maintain the normal structure and function of gingiva. All the components have a synergistic effect, so that gingival atrophy can be effectively prevented and improved.

Description

Technical Field

[0001] The present invention belongs to the technical field of biomedicine, and in particular relates to a pharmaceutical composition for preventing and treating gingival atrophy, and a preparation method and application thereof. Background Art

[0002] Gum recession is a common oral disease, and the occurrence of gum recession has complex reasons. First of all, periodontitis is one of the main factors leading to gum recession. Due to the proliferation of bacteria in the mouth, dental plaque and tartar are formed, which stimulate the gingival tissue and trigger an inflammatory response. Long-term periodontitis can lead to periodontal tissue destruction and alveolar bone absorption, which in turn causes gum recession. Secondly, improper brushing methods can also cause gum recession. For example, using a too hard toothbrush, brushing horizontally, etc., can cause mechanical damage to the gums, leading to gum recession over time. In addition, aging, genetic factors, poor restorations, etc. are also closely related to the occurrence of gum recession.

[0003] Gum recession can bring about a series of serious consequences. On the one hand, it affects aesthetics. After gum recession, the roots of the teeth are exposed, the teeth appear longer, and the gaps between the teeth become larger, which has a negative impact on the patient's facial appearance. On the other hand, gum recession can make teeth sensitive. Exposed tooth roots are more sensitive to stimuli such as hot, cold, sour, and sweet, and patients will feel pain and discomfort when eating or brushing their teeth. What's more serious is that gum recession increases the risk of loose and falling teeth. As gum recession worsens, the alveolar bone is continuously absorbed, the teeth lose the support of the surrounding tissues, gradually become loose, and can eventually lead to tooth loss.

[0004] At present, the clinical treatment of gingival recession mainly includes basic treatment, surgical treatment and drug treatment. Basic treatment includes oral hygiene guidance, cleaning, scaling, etc., which aims to remove local irritants and control inflammation. However, these methods are often difficult to completely restore gingival recession that has already occurred. Surgical treatments such as mucogingival surgery, although they can improve the condition of gingival recession to a certain extent, have high surgical risks and relatively low patient acceptance. Drug treatment is relatively mild. Some drugs that have the function of promoting gingival tissue regeneration can stimulate the proliferation and differentiation of gingival cells and promote the synthesis of collagen, thereby helping to restore the normal morphology and function of the gums. In addition, there are some drugs that can inhibit inflammatory responses, reduce the damage to periodontal tissues, and thus slow down the process of gingival recession.

[0005] Although there are some oral medications, topical medications and medications added to toothpaste for the prevention and treatment of gingival atrophy, these medications still have many shortcomings.

[0006] (1) Limited efficacy: Existing drugs are often not ideal in promoting gum tissue regeneration. The treatment course is long, the effect is slow, and the effect is not obvious. Patients need to take medication for a long time, which not only increases the difficulty of patient compliance, but also brings inconvenience to patients' lives.

[0007] (2) Risk of side effects: Some drugs used to prevent and treat gum atrophy may cause side effects. For example, some anti-inflammatory drugs may cause adverse reactions such as gastrointestinal discomfort, dizziness, and rash. Long-term use of these drugs may also cause certain damage to liver and kidney function.

[0008] Therefore, in order to better meet the needs of patients, it is necessary to further strengthen the research on gum atrophy drugs and develop safer and more effective drugs for the prevention and treatment of gum atrophy. Summary of the invention

[0009] In order to further improve the efficacy and safety of drugs for preventing and treating gum atrophy, the present invention provides a drug composition for preventing and treating gum atrophy, and its preparation method and application. The composition contains a specific proportion of eel peptide, bonito elastin peptide, quinoa rice extract, saponin rice extract, calendula extract, chamomile extract, chitosan, peach gum, hydroxyapatite, guar gum, soybean lecithin and other ingredients; the combination of the ingredients can quickly and efficiently repair the gum state and effectively delay the degree of gum atrophy. The specific technical scheme is as follows:

[0010] A pharmaceutical composition for preventing and treating gingival atrophy, the composition comprising the following raw materials in parts by weight: 0.1 to 0.5 parts of yellow eel peptide, 0.05 to 0.1 parts of bonito elastin peptide, 1 to 4 parts of quinoa rice extract, 3 to 8 parts of sapodilla rice extract, 0.5 to 3 parts of calendula extract, 0.5 to 3 parts of chamomile extract, 0.1 to 0.5 parts of chitosan, 0.2 to 0.5 parts of peach gum, 1 to 5 parts of hydroxyapatite, 0.05 to 0.08 parts of guar gum, 0.1 to 0.3 parts of soybean lecithin, the balance being water, and the water content of the composition being 10% to 25%; the yellow eel peptide is yellow eel preserved egg The white is hydrolyzed by trypsin and collagenase IV, and then by nattokinase and chymotrypsin to obtain products below 3kDa; the bonito elastin peptide is the product below 3kDa obtained by hydrolyzing the bonito heart arterial bulb with porcine pancreatic elastase; the quinoa rice extract is the product below 3kDa obtained by hydrolyzing quinoa rice with amylase, pectinase and hemicellulase, and then by ficin and nattokinase in sequence; the saponin rice extract is the product below 3kDa obtained by hydrolyzing saponin rice with amylase, pectinase and hemicellulase, and then by papain and subtilisin in sequence.

[0011] In the above-mentioned composition, the preparation method of the eel peptide comprises the following steps: taking the eel skin, washing it, soaking it in warm water, freezing it to remove acid, thawing it, washing it with water, draining it, and crushing it into a slurry to obtain fish skin slurry; adding a saline solution with a mass of 10 to 12 times the mass of the fish skin slurry to the fish skin slurry, stirring and soaking it, centrifuging it, collecting the supernatant, adding an ammonium sulfate aqueous solution to the supernatant, continuously stirring until the protein is completely precipitated, centrifuging it, collecting the precipitate, and obtaining the eel skin protein; dispersing the eel skin protein in 10 to 12 times the mass of the eel skin; In the water with high protein content, the pH is adjusted to 7.4-7.8, trypsin and collagenase IV are added, and the enzymatic hydrolysis is carried out at 36℃-38℃ for 50min-90min, the enzyme is inactivated by heating, and the pH is adjusted to 8.0-8.5 after the pH is reduced to 36℃-38℃, nattokinase and chymotrypsin are added, and the enzymatic hydrolysis is carried out at 36℃-38℃ for 50min-70min, the enzyme is inactivated by heating, and the pH is reduced to room temperature, and the filtrate is filtered through a sieve, and the filtrate is filtered through a 3kDa ultrafiltration membrane, and the ultrafiltrate below 3kDa is taken and freeze-dried to obtain eel peptide.

[0012] In the above-mentioned preparation method of eel peptide, the temperature of warm water immersion is 40℃~45℃, and the time of warm water immersion is 30min~40min; the temperature of frozen acid removal is -15℃~-25℃, and the time of frozen acid removal is 2h~3h; the concentration of saline solution is 0.1mol / L~0.2mol / L; the speed of stirring and immersing is 10r / min~30r / min, and the time of stirring and immersing is 3h~4h; the concentration of ammonium sulfate aqueous solution is 4.0mol / L~5.0mol / L; the speed of continuous stirring is 10r / min~30r / min; the speed of centrifugal separation is 4000r / min~6000r / min, and the time of centrifugal separation is 10min~30min; the added amount of trypsin and collagenase IV is 0.5%~1.5% of the mass of eel skin protein; the added amount of nattokinase and chymotrypsin is 0.8%~2% of the mass of eel skin protein.

[0013] In the above-mentioned composition, the preparation method of bonito elastin peptide comprises the following steps: taking bonito heart artery bulb, washing, and crushing into slurry, adding water 10 to 12 times the mass of the slurry into the slurry, adjusting the pH value to 7.8 to 8.5, adding 0.5% to 2% of porcine pancreatic elastase by mass of the bonito heart artery bulb, performing enzymolysis at 25 to 28°C for 2 to 3 hours, heating to inactivate the enzyme, cooling to room temperature, filtering with a sieve, taking the filtrate, filtering with a 3kDa ultrafiltration membrane, taking the ultrafiltrate below 3kDa, and freeze-drying to obtain the bonito elastin peptide.

[0014] In the above composition, the preparation method of quinoa rice extract comprises the following steps: taking quinoa rice, adding water of 4 to 6 times the mass of quinoa rice, soaking for 40 to 60 minutes, filtering to remove water, crushing, then adding water of 8 to 12 times the mass of quinoa rice, adjusting the pH to 4.0 to 5.0, adding 0.5% to 2% of amylase, pectinase and hemicellulase by mass of quinoa rice, enzymolysis at 37°C to 45°C for 2h to 4h, adjusting the pH to 5.5 to 6.0, adding 0. 5% to 1.5% of ficin is enzymatically hydrolyzed at 60 to 65° C. for 60 to 90 minutes, the temperature is raised to inactivate the enzyme, the pH is adjusted to 8.0 to 8.5 after the temperature is lowered to 36 to 38° C., 0.5% to 1.5% of nattokinase by weight of quinoa rice is added, the temperature is raised to inactivate the enzyme, the temperature is lowered to room temperature, sieve is used for filtering, the filtrate is taken, a 3 kDa ultrafiltration membrane is used for filtering, the ultrafiltrate below 3 kDa is taken, and freeze-drying is performed to obtain a quinoa rice extract.

[0015] In the above composition, the preparation method of the sapodilla rice extract comprises the following steps: taking sapodilla rice and adding it to water of 4 to 6 times the mass of sapodilla rice, soaking for 40 to 60 minutes, filtering to remove water, crushing, and then adding water of 8 to 12 times the mass of sapodilla rice, adjusting the pH to 4.0 to 5.0, adding 0.5% to 2% of amylase, pectinase and hemicellulase by mass of sapodilla rice, enzymolysis at 37°C to 45°C for 2h to 4h, adjusting the pH to 5.7 to 6.5, adding 0.5% by mass of sapodilla rice, and The invention discloses a method for preparing a sapodilla rice extract according to the present invention. The method comprises the following steps: adding papain of 0.5% to 1.5% by weight of papain, performing enzymolysis at 55°C to 60°C for 60min to 90min, heating to inactivate the enzyme, reducing the temperature to 50°C to 60°C, adjusting the pH value to 8.0 to 9.0, adding subtilisin of 0.5% to 1.5% by weight of sapodilla rice, performing enzymolysis at 50°C to 60°C for 50min to 90min, heating to inactivate the enzyme, reducing the temperature to room temperature, filtering with a sieve, taking the filtrate, filtering with a 3kDa ultrafiltration membrane, taking the ultrafiltrate below 3kDa, and freeze-drying the extract to obtain a sapodilla rice extract.

[0016] In the above technical solution, the temperature for heating and inactivating the enzyme is 80°C to 90°C, the time for heating and inactivating the enzyme is 15min to 25min; the mesh size of the sieve is 250 to 325 meshes.

[0017] In the above composition, the calendula extract is an ethanol percolation extract. The preparation method of the calendula extract comprises the following steps: taking the flowers of the calendula, crushing them to obtain a crushed material, using ethanol with a mass of 6 to 8 times that of the flowers to percolate the crushed material to obtain a percolate, concentrating, and drying to obtain the calendula extract.

[0018] In the above composition, the chamomile extract is an ethanol percolation extract. The preparation method of the chamomile extract comprises the following steps: taking chamomile flowers, crushing them to obtain a crushed material, using ethanol with an amount of 6 to 8 times the mass of the flowers to percolate the crushed material to obtain a percolate, concentrating, and drying to obtain the chamomile extract.

[0019] The preparation method of the above-mentioned pharmaceutical composition for preventing and treating gingival atrophy comprises the following steps:

[0020] S1: Mix eel peptide, bonito elastin peptide, quinoa rice extract, sapodilla rice extract, calendula extract and chamomile extract according to weight percentage to obtain a mixture A;

[0021] S2: adding water of twice the mass of mixture A to mixture A, mixing and dispersing the mixture evenly to obtain a mixture B;

[0022] S3: crushing peach gum according to mass fraction, adding peach gum into water at a mass ratio of peach gum: water = 1: (3-5) to soak for 6h-10h to obtain a foamed material C;

[0023] S4: taking guar gum according to mass fraction, dissolving the guar gum in water at a mass ratio of guar gum: water = 1: (100-120) to obtain a glue solution D;

[0024] S5: Mix the mixed solution B, foaming material C, glue solution D and soybean lecithin in proportions by mass, then add chitosan and hydroxyapatite, mix well, and concentrate under negative pressure below 55° C. to a water content of 10% to 25% to obtain a composition.

[0025] The above-mentioned pharmaceutical composition for preventing and treating gum atrophy is used for preparing a medicament for coating the gum area for preventing and treating gum atrophy.

[0026] The pharmaceutical composition for preventing and treating gingival atrophy provided by the present invention, as well as its preparation method and application, has the following beneficial effects compared with the prior art:

[0027] 1. The composition of the present invention contains specific proportions of eel peptide, bonito elastin peptide, quinoa rice extract, saponin rice extract, calendula extract, chamomile extract, chitosan, peach gum, hydroxyapatite, guar gum, soy lecithin and other ingredients. The combination of the ingredients can quickly and efficiently repair the gum state and effectively delay the degree of gum atrophy.

[0028] 2. In the composition of the present invention, the eel peptide is a product below 3kDa obtained by co-enzymatic hydrolysis of eel skin protein with trypsin and collagenase IV, followed by co-enzymatic hydrolysis with nattokinase and chymotrypsin. The eel peptide obtained after specific enzymatic hydrolysis of eel skin protein contains a variety of biologically active ingredients. It helps maintain the normal structure and function of gingival tissue and prevent gingival atrophy by promoting the metabolism of gingival tissue cells, providing nutrients to cells, and enhancing cell vitality and repair ability. Its molecular weight is below 3kDa, which makes it easier to be absorbed and utilized by the human body and transdermally to play a biological role.

[0029] 3. In the composition of the present invention, the bonito elastin peptide is a product below 3 kDa obtained by enzymatic hydrolysis of bonito heart arterial bulb with porcine pancreatic elastase. The bonito elastin peptide obtained by enzymatic hydrolysis of bonito heart arterial bulb can provide elastic support for gingival tissue. Collagen and elastin in gingival tissue are essential for maintaining the elasticity and firmness of the gums. This ingredient can help replenish and maintain the levels of these proteins and prevent the gums from shrinking due to loss of elasticity. Similarly, a molecular weight below 3 kDa is conducive to absorption and can better play its supporting role for gingival tissue.

[0030] Fourth, in the composition of the present invention, the quinoa rice extract is a product below 3 kDa obtained by hydrolyzing quinoa rice with amylase, pectinase and hemicellulase, and then hydrolyzing it step by step with ficin and nattokinase. The quinoa rice extract obtained after a series of enzymatic hydrolysis contains a variety of nutrients and biological polypeptide active substances. It can regulate the immune function of gingival tissue, reduce inflammatory response, and promote the repair and regeneration of gingival tissue. At the same time, the nutrients it provides also help maintain the normal metabolism of gingival cells.

[0031] 5. In the composition of the present invention, the extract of sapodilla rice is a product of less than 3 kDa obtained by hydrolyzing sapodilla rice with amylase, pectinase and hemicellulase, and then hydrolyzing it step by step with papain and subtilisin. After specific enzymolysis, the sapodilla rice extract contains certain specific small molecule peptides and various small molecule substances, which can have antioxidant, anti-inflammatory and cell proliferation promoting effects. It can protect gingival tissue from free radical damage, reduce inflammatory response, promote the growth and repair of gingival cells, and thus help prevent gingival atrophy.

[0032] 6. In the composition of the present invention, the calendula extract is an ethanol percolation extract. The calendula ethanol percolation extract has anti-inflammatory, antibacterial and wound healing effects. In terms of preventing and treating gingival atrophy, it can reduce gingival inflammation, prevent bacterial infection, promote the healing of damaged gingival tissue, and maintain the healthy state of the gums.

[0033] 7. In the composition of the present invention, the chamomile extract is an ethanol percolation extract. The chamomile extract also has anti-inflammatory, soothing and antioxidant effects. It can relieve gingival inflammation and discomfort, reduce oxidative stress damage to gingival tissue, and help maintain the normal function of the gums.

[0034] 8. Adding chitosan to the composition of the present invention has antibacterial, wound healing and hemostatic effects. It can inhibit the growth of bacteria in the oral cavity, reduce the risk of gum infection, promote the healing of gum wounds, and prevent gum atrophy caused by infection and bleeding.

[0035] 9. Peach gum is added to the composition of the present invention, which has the effects of moisturizing, lubricating and promoting cell adhesion. It can maintain the moisture of gingival tissue and reduce the risk of gingival dryness and atrophy. At the same time, it promotes the adhesion of cells on gingival tissue and helps to maintain the integrity of gingival tissue. In addition, the combination of chitosan and peach gum can effectively improve the adhesion and moisturizing properties of the composition on teeth and gums, improve the drug absorption efficiency and prolong the time, thereby improving the improvement effect of gingival atrophy in short-term use and shortening the course of treatment.

[0036] 10. Hydroxyapatite solid filler is added to the composition of the present invention. Hydroxyapatite can provide mineral support for the gums in preventing and treating gum atrophy, enhance the hardness and stability of the gums, and help maintain the normal structure and function of the gums. Hydroxyapatite can serve as a carrier to improve the adhesion of active substances and prevent the rapid loss of drug components.

[0037] 11. Adding guar gum to the composition of the present invention can increase the viscosity of the pharmaceutical composition, make it stay in the oral cavity for a longer time, and increase the contact time and effect of each component with the gum tissue. At the same time, guar gum can also have a certain moisturizing and lubricating effect.

[0038] 12. The addition of soybean lecithin in the composition of the present invention has emulsifying, moisturizing and nourishing effects. It can promote the dispersion and absorption of other ingredients, maintain the moisture of gingival tissue, provide nutrients for gingival cells, and help maintain the healthy state of gums.

[0039] 13. The combination of eel peptide, bonito elastin peptide, quinoa rice extract, and saponin rice extract can work together to promote the repair and regeneration of gingival tissue. The combination of calendula extract, chamomile extract, and chitosan has anti-inflammatory and antibacterial effects, and can work synergistically to reduce gingival inflammation and prevent bacterial infection. The combination of chitosan and calendula extract has the effect of promoting wound healing, and can work synergistically to accelerate the repair of damaged gingival tissue. The combination of eel peptide, bonito elastin peptide, and hydroxyapatite can provide nutrition and support for gingival tissue and jointly maintain the normal structure and function of the gums. Hydroxyapatite provides mineral support, and eel peptide and bonito elastin peptide provide protein nutrition. The synergistic effect can enhance the hardness and elasticity of the gums and prevent gum atrophy. DETAILED DESCRIPTION

[0040] The present invention is further described below in conjunction with specific implementation cases, but the present invention is not limited to these embodiments.

[0041] Example 1

[0042] A pharmaceutical composition for preventing and treating gum atrophy, the composition comprising the following raw materials in parts by mass: 0.3 parts of yellow eel peptide, 0.08 parts of bonito elastin peptide, 2.5 parts of quinoa rice extract, 5 parts of sapodilla rice extract, 2 parts of calendula extract, 2 parts of chamomile extract, 0.3 parts of chitosan, 0.3 parts of peach gum, 3 parts of hydroxyapatite, 0.06 parts of guar gum, 0.2 parts of soybean lecithin, and the balance being water, and the water content of the composition is 15%.

[0043] The preparation method of eel peptide comprises the following steps: taking eel skin, washing it, soaking it in 42°C warm water for 35 minutes, freezing it at -20°C to remove acid for 2.5 hours, thawing it, washing it with water, draining it, and crushing it into a slurry to obtain fish skin slurry; adding a 0.15 mol / L saline solution with a concentration of 11 times the mass of the fish skin slurry to the fish skin slurry, stirring and soaking it for 3.5 hours at 20r / min, centrifuging it at 5000r / min for 20 minutes, collecting the supernatant, adding a 4.5 mol / L ammonium sulfate aqueous solution to the supernatant, stirring it continuously at 20r / min until the protein is completely precipitated, centrifuging it at 5000r / min for 20 minutes, collecting it, Precipitation to obtain yellow eel skin protein; dispersing the yellow eel skin protein in water with 11 times the amount of the yellow eel skin protein, adjusting the pH to 7.6, adding trypsin and collagenase IV with a mass of 1.0% of the yellow eel skin protein, enzymolysis at 37°C for 70 minutes, heating to 85°C for 20 minutes to inactivate the enzyme, and after cooling to 37°C, adjusting the pH to 8.2, adding nattokinase and chymotrypsin with a mass of 1.4% of the yellow eel skin protein, enzymolysis at 37°C for 60 minutes, heating to 85°C for 20 minutes to inactivate the enzyme, and after cooling to room temperature, filtering with a 325-mesh screen, taking the filtrate, filtering with a 3kDa ultrafiltration membrane, taking the ultrafiltrate below 3kDa, and freeze-drying to obtain yellow eel peptide.

[0044] The preparation method of bonito elastin peptide comprises the following steps: taking bonito heart artery bulb, washing, crushing into slurry, adding water of 11 times the mass of the slurry into the slurry, adjusting the pH to 8.1, adding porcine pancreatic elastase of 1.4% of the mass of bonito heart artery bulb, performing enzymolysis at 26 DEG C for 2.5 hours, heating to 85 DEG C for 20 minutes to inactivate the enzyme, filtering with a 325-mesh screen after cooling to room temperature, taking the filtrate, filtering with a 3kDa ultrafiltration membrane, taking the ultrafiltrate below 3kDa, and freeze-drying to obtain the bonito elastin peptide.

[0045] The preparation method of quinoa rice extract comprises the following steps: taking quinoa rice, adding water of 5 times the mass of quinoa rice, soaking for 50 minutes, filtering to remove water, crushing, then adding water of 10 times the mass of quinoa rice, adjusting pH to 4.5, adding amylase, pectinase and hemicellulase of 1.4% by mass of quinoa rice, enzymolysis at 42°C for 3 hours, adjusting pH to 5.8, adding ficin of 1.0% by mass of quinoa rice, enzymolysis at 62°C for 75 minutes, heating to 85°C to inactivate enzyme for 20 minutes, reducing the temperature to 37°C, adjusting pH to 8.2, adding nattokinase of 1.0% by mass of quinoa rice, enzymolysis at 37°C for 75 minutes, heating to 85°C to inactivate enzyme for 20 minutes, reducing the temperature to room temperature, filtering with a 325-mesh screen, taking the filtrate, filtering with a 3kDa ultrafiltration membrane, taking the ultrafiltrate below 3kDa, freeze-drying, and obtaining the quinoa rice extract.

[0046] The preparation method of the sapodilla rice extract comprises the following steps: adding sapodilla rice to water with a mass of 5 times the mass of the sapodilla rice, soaking for 50 minutes, filtering to remove water, crushing, then adding water with a mass of 10 times the mass of the sapodilla rice, adjusting the pH to 4.5, adding 1.5% of amylase, pectinase and hemicellulase by mass of the sapodilla rice, enzymolysis at 40°C for 3 hours, adjusting the pH to 6.0, adding 1.0% of papain by mass of the sapodilla rice, enzymolysis at 58°C for 75 minutes, heating to 85°C to inactivate the enzyme for 20 minutes, cooling to 55°C, adjusting the pH to 8.5, adding 1.0% of subtilisin by mass of the sapodilla rice, enzymolysis at 55°C for 75 minutes, heating to 85°C to inactivate the enzyme for 20 minutes, cooling to room temperature, filtering with a 325-mesh screen, taking the filtrate, filtering with a 3kDa ultrafiltration membrane, taking the ultrafiltrate below 3kDa, freeze-drying, and obtaining the sapodilla rice extract.

[0047] The calendula extract is an ethanol percolation extract, and the preparation method of the calendula extract comprises the following steps: taking the flowers of the calendula, crushing them to obtain a crushed material, using ethanol with a mass of 7 times that of the flowers to percolate the crushed material to obtain a percolate, concentrating, and drying to obtain the calendula extract.

[0048] Among them, the chamomile extract is an ethanol percolation extract, and the preparation method of the chamomile extract includes the following steps: taking chamomile flowers, crushing to obtain a crushed material, using ethanol 7 times the mass of the flowers to percolate the crushed material to obtain a percolate, concentrating, and drying to obtain a chamomile extract.

[0049] The preparation method of the above-mentioned pharmaceutical composition for preventing and treating gingival atrophy comprises the following steps:

[0050] S1: Mix eel peptide, bonito elastin peptide, quinoa rice extract, sapodilla rice extract, calendula extract and chamomile extract according to weight percentage to obtain a mixture A;

[0051] S2: Add water twice the mass of mixture A to mixture A, mix and disperse evenly to obtain mixed solution B;

[0052] S3: crushing peach gum according to mass fraction, adding peach gum into water at a mass ratio of peach gum: water = 1:4, and soaking for 8 hours to obtain a foamed material C;

[0053] S4: taking guar gum according to mass fraction, dissolving the guar gum in water at a mass ratio of guar gum: water = 1:110, to obtain a glue solution D;

[0054] S5: Mix the mixed solution B, foaming material C, glue solution D and soybean lecithin in proportions by mass, then add chitosan and hydroxyapatite, mix well, and concentrate under negative pressure at 54° C. to a water content of 15% to obtain a composition.

[0055] The above-mentioned pharmaceutical composition for preventing and treating gum atrophy is used for preparing a medicament for coating the gum area for preventing and treating gum atrophy.

[0056] Example 2

[0057] A pharmaceutical composition for preventing and treating gingival atrophy, the composition comprising the following raw materials in parts by mass: 0.1 parts of yellow eel peptide, 0.05 parts of bonito elastin peptide, 1 part of quinoa rice extract, 3 parts of sapodilla rice extract, 0.5 parts of calendula extract, 0.5 parts of chamomile extract, 0.1 parts of chitosan, 0.2 parts of peach gum, 1 part of hydroxyapatite, 0.05 parts of guar gum, 0.1 parts of soybean lecithin, and the balance being water, and the water content of the composition is 10%.

[0058] The preparation method of eel peptide comprises the following steps: taking eel skin, washing it, soaking it in 40°C warm water for 30 minutes, freezing it at -15°C to remove acid for 2 hours, thawing it, washing it with water, draining it, and crushing it into a slurry to obtain fish skin slurry; adding a 0.1 mol / L saline solution with a concentration of 10 times the mass of the fish skin slurry to the fish skin slurry, stirring and soaking it at 10r / min for 3 hours, centrifuging it at 4000r / min for 10 minutes, collecting the supernatant, adding a 4.0 mol / L ammonium sulfate aqueous solution to the supernatant, stirring it continuously at 10r / min until the protein is completely precipitated, centrifuging it at 4000r / min for 10 minutes, collecting the precipitate, Obtaining yellow eel skin protein; dispersing the yellow eel skin protein in water with an amount 10 times that of the yellow eel skin protein, adjusting the pH to 7.4, adding trypsin and collagenase IV with an amount of 0.5% of the yellow eel skin protein, performing enzymatic hydrolysis at 36°C for 50 minutes, heating to 80°C for 25 minutes to inactivate the enzymes, adjusting the pH to 8.0 after the temperature drops to 36°C, adding nattokinase and chymotrypsin with an amount of 0.8% of the yellow eel skin protein, performing enzymatic hydrolysis at 36°C for 50 minutes, heating to 80°C for 25 minutes to inactivate the enzymes, cooling the temperature to room temperature, filtering with a 250-mesh screen, taking the filtrate, filtering with a 3 kDa ultrafiltration membrane, taking the ultrafiltrate below 3 kDa, and freeze-drying to obtain yellow eel peptide.

[0059] The preparation method of bonito elastin peptide comprises the following steps: taking bonito heart artery bulb, washing, crushing into slurry, adding water 10 times the mass of the slurry into the slurry, adjusting the pH to 7.8, adding 0.5% of porcine pancreatic elastase by mass of the bonito heart artery bulb, performing enzymolysis at 25 DEG C for 2 hours, heating to 80 DEG C for 25 minutes to inactivate the enzyme, cooling to room temperature, filtering with a 250-mesh screen, taking the filtrate, filtering with a 3 kDa ultrafiltration membrane, taking the ultrafiltrate below 3 kDa, and freeze-drying to obtain the bonito elastin peptide.

[0060] The preparation method of quinoa rice extract comprises the following steps: taking quinoa rice, adding water of 4 times the mass of quinoa rice, soaking for 40 minutes, filtering to remove water, crushing, then adding water of 8 times the mass of quinoa rice, adjusting pH 4.0, adding 0.5% of amylase, pectinase and hemicellulase by mass of quinoa rice, enzymolysis at 37°C for 2 hours, adjusting pH 5.5, adding 0.5% of fig protease by mass of quinoa rice, enzymolysis at 60°C for 60 minutes, heating to 80°C to inactivate enzyme for 25 minutes, reducing the temperature to 36°C, adjusting pH 8.0, adding 0.5% of nattokinase by mass of quinoa rice, enzymolysis at 36°C for 60 minutes, heating to 80°C to inactivate enzyme for 25 minutes, reducing the temperature to room temperature, filtering with a 250-mesh screen, taking the filtrate, filtering with a 3kDa ultrafiltration membrane, taking the ultrafiltrate below 3kDa, freeze-drying, and obtaining the quinoa rice extract.

[0061] The preparation method of the sapodilla rice extract comprises the following steps: adding sapodilla rice to water with a mass of 4 times the mass of the sapodilla rice, soaking for 40 minutes, filtering to remove water, crushing, then adding water with a mass of 8 times the mass of the sapodilla rice, adjusting the pH to 4.0, adding 0.5% of amylase, pectinase and hemicellulase by mass of the sapodilla rice, enzymolysis at 37°C for 2 hours, adjusting the pH to 5.7, adding 0.5% of papain by mass of the sapodilla rice, enzymolysis at 55°C for 60 minutes, heating to 80°C to inactivate the enzyme for 25 minutes, cooling to 50°C, adjusting the pH to 8.0, adding 0.5% of subtilisin by mass of the sapodilla rice, enzymolysis at 50°C for 50 minutes, heating to 80°C to inactivate the enzyme for 25 minutes, cooling to room temperature, filtering with a 250-mesh screen, taking the filtrate, filtering with a 3kDa ultrafiltration membrane, taking the ultrafiltrate below 3kDa, freeze-drying, and obtaining the sapodilla rice extract.

[0062] The calendula extract is an ethanol percolation extract, and the preparation method of the calendula extract comprises the following steps: taking the flowers of the calendula, crushing them to obtain a crushed material, using ethanol 6 times the mass of the flowers to percolate the crushed material to obtain a percolate, concentrating, and drying to obtain the calendula extract.

[0063] Among them, the chamomile extract is an ethanol percolation extract, and the preparation method of the chamomile extract includes the following steps: taking chamomile flowers, crushing to obtain a crushed material, using ethanol 6 times the mass of the flowers to percolate the crushed material to obtain a percolate, concentrating, and drying to obtain a chamomile extract.

[0064] The preparation method of the above-mentioned pharmaceutical composition for preventing and treating gingival atrophy comprises the following steps:

[0065] S1: Mix eel peptide, bonito elastin peptide, quinoa rice extract, sapodilla rice extract, calendula extract and chamomile extract according to weight percentage to obtain a mixture A;

[0066] S2: Add 2.5 times the mass of water of mixture A to mixture A, mix and disperse evenly, and obtain mixed solution B;

[0067] S3: crushing peach gum according to mass fraction, adding peach gum into water at a mass ratio of peach gum: water = 1:3.5, and soaking for 7 hours to obtain a foamed material C;

[0068] S4: taking guar gum according to mass fraction, dissolving the guar gum in water at a mass ratio of guar gum: water = 1:100, to obtain a glue solution D;

[0069] S5: Mix the mixed solution B, foaming material C, glue solution D and soybean lecithin in proportions by mass, then add chitosan and hydroxyapatite, mix well, and concentrate under negative pressure at 52° C. to a water content of 10% to obtain a composition.

[0070] The above-mentioned pharmaceutical composition for preventing and treating gum atrophy is used for preparing a medicament for coating the gum area for preventing and treating gum atrophy.

[0071] Example 3

[0072] A pharmaceutical composition for preventing and treating gingival atrophy, the composition comprising the following raw materials in parts by mass: 0.1 parts of yellow eel peptide, 0.1 parts of bonito elastin peptide, 1 parts of quinoa rice extract, 8 parts of sapodilla rice extract, 0.5 parts of calendula extract, 3 parts of chamomile extract, 0.1 parts of chitosan, 0.5 parts of peach gum, 1 part of hydroxyapatite, 0.08 parts of guar gum, 0.1 parts of soybean lecithin, and the balance being water, and the water content of the composition is 12%.

[0073] The preparation method of eel peptide comprises the following steps: taking eel skin, washing it, soaking it in 45°C warm water for 30 minutes, freezing it at -25°C to remove acid for 2 hours, thawing it, washing it with water, draining it, and crushing it into a slurry to obtain fish skin slurry; adding a 0.1 mol / L saline solution with a concentration of 12 times the mass of the fish skin slurry to the fish skin slurry, stirring and soaking it at 30r / min for 3 hours, centrifuging it at 6000r / min for 10 minutes, collecting the supernatant, adding a 5.0 mol / L ammonium sulfate aqueous solution to the supernatant, stirring it continuously at 10r / min until the protein is completely precipitated, centrifuging it at 6000r / min for 10 minutes, collecting the precipitate, Obtaining yellow eel skin protein; dispersing the yellow eel skin protein in water with an amount 12 times that of the yellow eel skin protein, adjusting the pH to 7.4, adding trypsin and collagenase IV with an amount of 1.5% of the yellow eel skin protein, performing enzymatic hydrolysis at 36°C for 90 minutes, heating to 90°C for 15 minutes to inactivate the enzymes, adjusting the pH to 8.5 after the temperature drops to 36°C, adding nattokinase and chymotrypsin with an amount of 0.8% of the yellow eel skin protein, performing enzymatic hydrolysis at 38°C for 50 minutes, heating to 90°C for 15 minutes to inactivate the enzymes, cooling the temperature to room temperature, filtering with a 325-mesh screen, taking the filtrate, filtering with a 3kDa ultrafiltration membrane, taking the ultrafiltrate below 3kDa, and freeze-drying to obtain yellow eel peptide.

[0074] The preparation method of bonito elastin peptide comprises the following steps: taking bonito heart artery bulb, washing it, crushing it into slurry, adding water 10 times the mass of the slurry into the slurry, adjusting the pH to 8.5, adding 0.5% of porcine pancreatic elastase by mass of the bonito heart artery bulb, performing enzymolysis at 28 DEG C for 2 hours, heating to 90 DEG C for 15 minutes to inactivate the enzyme, filtering with a 325-mesh screen after cooling to room temperature, taking the filtrate, filtering with a 3kDa ultrafiltration membrane, taking the ultrafiltrate below 3kDa, and freeze-drying to obtain the bonito elastin peptide.

[0075] The preparation method of quinoa rice extract comprises the following steps: taking quinoa rice, adding water of 4 times the mass of quinoa rice, soaking for 60 minutes, filtering to remove water, crushing, then adding water of 8 times the mass of quinoa rice, adjusting pH to 5.0, adding 0.5% of amylase, pectinase and hemicellulase by mass of quinoa rice, enzymolysis at 45°C for 2 hours, adjusting pH to 6.0, adding 0.5% of fig protease by mass of quinoa rice, enzymolysis at 65°C for 60 minutes, heating to 90°C to inactivate enzyme for 15 minutes, reducing the temperature to 38°C, adjusting pH to 8.0, adding 1.5% of nattokinase by mass of quinoa rice, enzymolysis at 36°C for 90 minutes, heating to 90°C to inactivate enzyme for 15 minutes, reducing the temperature to room temperature, filtering with a 250-mesh screen, taking the filtrate, filtering with a 3kDa ultrafiltration membrane, taking the ultrafiltrate below 3kDa, freeze-drying, and obtaining the quinoa rice extract.

[0076] The preparation method of the sapodilla rice extract comprises the following steps: adding sapodilla rice to water with a mass of 6 times the mass of the sapodilla rice, soaking for 40 minutes, filtering to remove water, crushing, then adding water with a mass of 12 times the mass of the sapodilla rice, adjusting the pH to 4.0, adding 2% of amylase, pectinase and hemicellulase by mass of the sapodilla rice, enzymolysis at 37°C for 4 hours, adjusting the pH to 5.7, adding 1.5% of papain by mass of the sapodilla rice, enzymolysis at 55°C for 90 minutes, heating to 90°C to inactivate the enzyme for 15 minutes, cooling to 50°C, adjusting the pH to 9.0, adding 0.5% of subtilisin by mass of the sapodilla rice, enzymolysis at 60°C for 50 minutes, heating to 90°C to inactivate the enzyme for 15 minutes, cooling to room temperature, filtering with a 325-mesh screen, taking the filtrate, filtering with a 3kDa ultrafiltration membrane, taking the ultrafiltrate below 3kDa, freeze-drying, and obtaining the sapodilla rice extract.

[0077] The calendula extract is an ethanol percolation extract, and the preparation method of the calendula extract comprises the following steps: taking the flowers of the calendula, crushing them to obtain a crushed material, using ethanol with an amount of 8 times the mass of the flowers to percolate the crushed material to obtain a percolate, concentrating, and drying to obtain the calendula extract.

[0078] Among them, the chamomile extract is an ethanol percolation extract, and the preparation method of the chamomile extract includes the following steps: taking chamomile flowers, crushing to obtain a crushed material, using ethanol with an amount of 8 times the mass of the flowers to percolate the crushed material to obtain a percolate, concentrating, and drying to obtain the chamomile extract.

[0079] The preparation method of the above-mentioned pharmaceutical composition for preventing and treating gingival atrophy comprises the following steps:

[0080] S1: Mix eel peptide, bonito elastin peptide, quinoa rice extract, sapodilla rice extract, calendula extract and chamomile extract according to weight percentage to obtain a mixture A;

[0081] S2: Add 3 times the mass of water of mixture A to mixture A, mix and disperse evenly to obtain mixed solution B;

[0082] S3: crushing peach gum according to mass fraction, adding peach gum into water at a mass ratio of peach gum: water = 1:4.5, and soaking for 9 hours to obtain a foamed material C;

[0083] S4: taking guar gum by mass, dissolving the guar gum in water at a mass ratio of guar gum: water = 1:120, to obtain a glue solution D;

[0084] S5: Mix the mixed solution B, foaming material C, glue solution D and soybean lecithin in proportions by mass, then add chitosan and hydroxyapatite, mix well, and concentrate under negative pressure at 52° C. to a water content of 12% to obtain a composition.

[0085] The above-mentioned pharmaceutical composition for preventing and treating gum atrophy is used for preparing a medicament for coating the gum area for preventing and treating gum atrophy.

[0086] Example 4

[0087] A pharmaceutical composition for preventing and treating gum atrophy, the composition comprising the following raw materials in parts by mass: 0.5 parts of yellow eel peptide, 0.1 parts of bonito elastin peptide, 4 parts of quinoa rice extract, 8 parts of sapodilla rice extract, 3 parts of calendula extract, 3 parts of chamomile extract, 0.5 parts of chitosan, 0.5 parts of peach gum, 5 parts of hydroxyapatite, 0.08 parts of guar gum, 0.3 parts of soybean lecithin, and the balance being water, and the water content of the composition is 20%.

[0088] The preparation method of eel peptide comprises the following steps: taking eel skin, washing it, soaking it in 45°C warm water for 40 minutes, freezing it at -25°C to remove acid for 3 hours, thawing it, washing it with water, draining it, and crushing it into a slurry to obtain fish skin slurry; adding a 0.2 mol / L saline solution with a concentration of 12 times the mass of the fish skin slurry to the fish skin slurry, stirring and soaking it for 4 hours at 30r / min, centrifuging it at 6000r / min for 30 minutes, collecting the supernatant, adding a 5.0 mol / L ammonium sulfate aqueous solution to the supernatant, stirring it continuously at 30r / min until the protein is completely precipitated, centrifuging it at 6000r / min for 30 minutes, and collecting the precipitate. , and obtain the yellow eel skin protein; disperse the yellow eel skin protein in water with 12 times the amount of the yellow eel skin protein, adjust the pH to 7.8, add trypsin and collagenase IV with a mass of 1.5% of the yellow eel skin protein, perform enzymatic hydrolysis at 38°C for 90 minutes, heat to 85°C to inactivate the enzyme for 15 minutes, and after cooling to 38°C, adjust the pH to 8.5, add nattokinase and chymotrypsin with a mass of 2% of the yellow eel skin protein, perform enzymatic hydrolysis at 38°C for 70 minutes, heat to 85°C to inactivate the enzyme for 15 minutes, and after cooling to room temperature, filter with a 325-mesh sieve, take the filtrate, filter with a 3kDa ultrafiltration membrane, take the ultrafiltrate below 3kDa, and freeze-dry to obtain the yellow eel peptide.

[0089] The preparation method of bonito elastin peptide comprises the following steps: taking bonito heart artery bulb, washing, crushing into slurry, adding water of 12 times the mass of the slurry into the slurry, adjusting the pH to 8.5, adding 2% of porcine pancreatic elastase of the mass of bonito heart artery bulb, performing enzymolysis at 28 DEG C for 3 hours, heating to 85 DEG C for 15 minutes to inactivate the enzyme, filtering with a 325-mesh screen after cooling to room temperature, taking the filtrate, filtering with a 3kDa ultrafiltration membrane, taking the ultrafiltrate below 3kDa, and freeze-drying to obtain bonito elastin peptide.

[0090] The preparation method of quinoa rice extract comprises the following steps: taking quinoa rice, adding water of 6 times the mass of quinoa rice, soaking for 60 minutes, filtering to remove water, crushing, then adding water of 12 times the mass of quinoa rice, adjusting pH to 5.0, adding 2% of amylase, pectinase and hemicellulase by mass of quinoa rice, enzymolysis at 45°C for 4 hours, adjusting pH to 6.0, adding 1.5% of ficin by mass of quinoa rice, enzymolysis at 65°C for 90 minutes, heating to 85°C for 15 minutes to inactivate the enzyme, cooling to 38°C, adjusting pH to 8.5, adding 1.5% of nattokinase by mass of quinoa rice, enzymolysis at 38°C for 90 minutes, heating to 85°C for 15 minutes to inactivate the enzyme, cooling to room temperature, filtering with a 325-mesh screen, taking the filtrate, filtering with a 3kDa ultrafiltration membrane, taking the ultrafiltrate below 3kDa, freeze-drying, and obtaining the quinoa rice extract.

[0091] The preparation method of the sapodilla rice extract comprises the following steps: adding sapodilla rice to water with a mass of 6 times that of the sapodilla rice, soaking for 60 minutes, filtering to remove water, crushing, then adding water with a mass of 12 times that of the sapodilla rice, adjusting the pH to 5.0, adding 2% of amylase, pectinase and hemicellulase by mass of the sapodilla rice, enzymolysis at 45°C for 4 hours, adjusting the pH to 6.5, adding 1.5% of papain by mass of the sapodilla rice, enzymolysis at 60°C for 90 minutes, heating to 85°C for 15 minutes to inactivate the enzyme, cooling to 60°C, adjusting the pH to 9.0, adding 1.5% of subtilisin by mass of the sapodilla rice, enzymolysis at 60°C for 90 minutes, heating to 85°C for 15 minutes to inactivate the enzyme, cooling to room temperature, filtering with a 325-mesh screen, taking the filtrate, filtering with a 3kDa ultrafiltration membrane, taking the ultrafiltrate below 3kDa, freeze-drying, and obtaining the sapodilla rice extract.

[0092] The calendula extract is an ethanol percolation extract, and the preparation method of the calendula extract comprises the following steps: taking the flowers of the calendula, crushing them to obtain a crushed material, using ethanol 6.5 times the mass of the flowers to percolate the crushed material to obtain a percolate, concentrating, and drying to obtain the calendula extract.

[0093] Among them, the chamomile extract is an ethanol percolation extract, and the preparation method of the chamomile extract includes the following steps: taking chamomile flowers, crushing to obtain a crushed material, using ethanol 7.5 times the mass of the flowers to percolate the crushed material to obtain a percolate, concentrating, and drying to obtain a chamomile extract.

[0094] The preparation method of the above-mentioned pharmaceutical composition for preventing and treating gingival atrophy comprises the following steps:

[0095] S1: Mix eel peptide, bonito elastin peptide, quinoa rice extract, sapodilla rice extract, calendula extract and chamomile extract according to weight percentage to obtain a mixture A;

[0096] S2: Add water twice the mass of mixture A to mixture A, mix and disperse evenly to obtain mixed solution B;

[0097] S3: crushing peach gum according to mass fraction, adding peach gum into water at a mass ratio of peach gum: water = 1:5, and soaking for 6 hours to obtain a foamed material C;

[0098] S4: taking guar gum according to mass fraction, dissolving the guar gum in water at a mass ratio of guar gum: water = 1:100, to obtain a glue solution D;

[0099] S5: Mix the mixed solution B, foaming material C, glue solution D and soybean lecithin in proportions by mass, then add chitosan and hydroxyapatite, mix well, and concentrate under negative pressure at 50° C. to a water content of 20% to obtain a composition.

[0100] The above-mentioned pharmaceutical composition for preventing and treating gum atrophy is used for preparing a medicament for coating the gum area for preventing and treating gum atrophy.

[0101] Example 5

[0102] A pharmaceutical composition for preventing and treating gum atrophy, the composition comprising the following raw materials in parts by mass: 0.5 parts of yellow eel peptide, 0.05 parts of bonito elastin peptide, 4 parts of quinoa rice extract, 3 parts of sapodilla rice extract, 3 parts of calendula extract, 0.5 parts of chamomile extract, 0.5 parts of chitosan, 0.2 parts of peach gum, 5 parts of hydroxyapatite, 0.05 parts of guar gum, 0.3 parts of soybean lecithin, and the balance being water, and the water content of the composition is 25%.

[0103] The preparation method of eel peptide comprises the following steps: taking eel skin, washing it, soaking it in 45°C warm water for 30 minutes, freezing it at -25°C to remove acid for 2 hours, thawing it, washing it with water, draining it, and crushing it into a slurry to obtain fish skin slurry; adding a 0.1 mol / L saline solution with a concentration of 12 times the mass of the fish skin slurry to the fish skin slurry, stirring and soaking it at 30r / min for 3 hours, centrifuging it at 6000r / min for 10 minutes, collecting the supernatant, adding a 5.0 mol / L ammonium sulfate aqueous solution to the supernatant, stirring it continuously at 10r / min until the protein is completely precipitated, centrifuging it at 6000r / min for 10 minutes, collecting the precipitate, Obtaining yellow eel skin protein; dispersing the yellow eel skin protein in water with an amount 12 times that of the yellow eel skin protein, adjusting the pH to 7.4, adding trypsin and collagenase IV with an amount of 1.5% of the yellow eel skin protein, performing enzymatic hydrolysis at 36°C for 90 minutes, heating to 80°C for 15 minutes to inactivate the enzymes, adjusting the pH to 8.5 after the temperature drops to 36°C, adding nattokinase and chymotrypsin with an amount of 0.8% of the yellow eel skin protein, performing enzymatic hydrolysis at 38°C for 50 minutes, heating to 80°C for 15 minutes to inactivate the enzymes, cooling the temperature to room temperature, filtering with a 325-mesh screen, taking the filtrate, filtering with a 3kDa ultrafiltration membrane, taking the ultrafiltrate below 3kDa, and freeze-drying to obtain yellow eel peptide.

[0104] The preparation method of bonito elastin peptide comprises the following steps: taking bonito heart artery bulb, washing, crushing into slurry, adding water 10 times the mass of the slurry into the slurry, adjusting the pH to 8.5, adding 0.5% of porcine pancreatic elastase by mass of the bonito heart artery bulb, performing enzymolysis at 28 DEG C for 2 hours, heating to 80 DEG C for 15 minutes to inactivate the enzyme, cooling to room temperature, filtering with a 325-mesh screen, taking the filtrate, filtering with a 3kDa ultrafiltration membrane, taking the ultrafiltrate below 3kDa, and freeze-drying to obtain the bonito elastin peptide.

[0105] The preparation method of quinoa rice extract comprises the following steps: taking quinoa rice, adding water of 4 times the mass of quinoa rice, soaking for 60 minutes, filtering to remove water, crushing, then adding water of 8 times the mass of quinoa rice, adjusting pH to 5.0, adding 0.8% of amylase, pectinase and hemicellulase by mass of quinoa rice, enzymolysis at 40°C for 2.5 hours, adjusting pH to 5.8, adding 0.8% of fig protease by mass of quinoa rice, enzymolysis at 62°C for 70 minutes, heating to 80°C to inactivate enzyme for 15 minutes, reducing the temperature to 37°C, adjusting pH to 8.4, adding 0.8% of nattokinase by mass of quinoa rice, enzymolysis at 37°C for 80 minutes, heating to 80°C to inactivate enzyme for 15 minutes, reducing the temperature to room temperature, filtering with a 325-mesh screen, taking the filtrate, filtering with a 3kDa ultrafiltration membrane, taking the ultrafiltrate below 3kDa, freeze-drying, and obtaining the quinoa rice extract.

[0106] The preparation method of the sapodilla rice extract comprises the following steps: adding sapodilla rice to water with a mass of 5 times the mass of the sapodilla rice, soaking for 55 minutes, filtering to remove water, crushing, then adding water with a mass of 11 times the mass of the sapodilla rice, adjusting the pH to 4.8, adding 1.5% of amylase, pectinase and hemicellulase by mass of the sapodilla rice, enzymolysis at 43°C for 3.5 hours, adjusting the pH to 6.2, adding 1.2% of papain by mass of the sapodilla rice, enzymolysis at 58°C for 80 minutes, heating to 80°C to inactivate the enzyme for 15 minutes, cooling to 52°C, adjusting the pH to 8.4, adding 0.8% of subtilisin by mass of the sapodilla rice, enzymolysis at 58°C for 80 minutes, heating to 80°C to inactivate the enzyme for 15 minutes, cooling to room temperature, filtering with a 325-mesh screen, taking the filtrate, filtering with a 3kDa ultrafiltration membrane, taking the ultrafiltrate below 3kDa, freeze-drying, and obtaining the sapodilla rice extract.

[0107] The calendula extract is an ethanol percolation extract, and the preparation method of the calendula extract comprises the following steps: taking the flowers of the calendula, crushing them to obtain a crushed material, using ethanol with an amount of 8 times the mass of the flowers to percolate the crushed material to obtain a percolate, concentrating, and drying to obtain the calendula extract.

[0108] Among them, the chamomile extract is an ethanol percolation extract, and the preparation method of the chamomile extract includes the following steps: taking chamomile flowers, crushing to obtain a crushed material, using ethanol 6 times the mass of the flowers to percolate the crushed material to obtain a percolate, concentrating, and drying to obtain a chamomile extract.

[0109] The preparation method of the above-mentioned pharmaceutical composition for preventing and treating gingival atrophy comprises the following steps:

[0110] S1: Mix eel peptide, bonito elastin peptide, quinoa rice extract, sapodilla rice extract, calendula extract and chamomile extract according to weight percentage to obtain a mixture A;

[0111] S2: Add 3 times the mass of water of mixture A to mixture A, mix and disperse evenly to obtain mixed solution B;

[0112] S3: crushing peach gum according to mass fraction, adding peach gum into water at a mass ratio of peach gum: water = 1:5, and soaking for 10 hours to obtain a foamed material C;

[0113] S4: taking guar gum by mass, dissolving the guar gum in water at a mass ratio of guar gum: water = 1:120, to obtain a glue solution D;

[0114] S5: Mix the mixed solution B, foaming material C, glue solution D and soybean lecithin in proportions by mass, then add chitosan and hydroxyapatite, mix well, and concentrate under negative pressure at 48° C. to a water content of 25% to obtain a composition.

[0115] The above-mentioned pharmaceutical composition for preventing and treating gum atrophy is used for preparing a medicament for coating the gum area for preventing and treating gum atrophy.

[0116] The enzyme activity of trypsin used in the above embodiments is 4000 U / g, the enzyme activity of collagenase IV is 100,000 U / g, the enzyme activity of nattokinase is 20,000 U / g, the enzyme activity of chymotrypsin is 500,000 U / g, the enzyme activity of porcine pancreatic elastase is 30,000 U / g, the enzyme activity of amylase is 20,000 U / g, the enzyme activity of pectinase is 30,000 U / g, the enzyme activity of hemicellulase is 10 U / g, the enzyme activity of ficin is 10,000 U / g, the enzyme activity of papain is 100,000 U / g, the bacterial activity of subtilisin is 20,000 U / g, and the particle size of hydroxyapatite is less than 5 μm.

[0117] Comparative Example 1

[0118] In the preparation method of the eel peptide, the eel skin protein is dispersed in water with 11 times the mass of the eel skin protein, the pH is adjusted to 7.6, and trypsin and collagenase IV with the mass of the eel skin protein being 1.0% are added, and the eel skin protein is hydrolyzed at 37°C for 70 minutes, and the enzyme is inactivated at 85°C for 20 minutes. After cooling to room temperature, the filtrate is filtered through a 325-mesh screen, and the filtrate is filtered through a 3kDa ultrafiltration membrane, and the ultrafiltrate below 3kDa is taken and freeze-dried to obtain the eel peptide. Other methods and parameters are the same as those in Example 1.

[0119] Comparative Example 2

[0120] In the preparation method of the eel peptide, the eel skin protein is dispersed in water with 11 times the amount of the eel skin protein, the pH is adjusted to 8.2, nattokinase and chymotrypsin with the amount of eel skin protein being 1.4% are added, the eel skin protein is hydrolyzed at 37°C for 60 minutes, the enzyme is inactivated at 85°C for 20 minutes, and after cooling to room temperature, the eel skin protein is filtered using a 325-mesh screen, the filtrate is filtered using a 3kDa ultrafiltration membrane, the ultrafiltrate below 3kDa is taken, and the eel peptide is obtained by freeze-drying. Other methods and parameters are the same as those in Example 1.

[0121] Comparative Example 3

[0122] No eel peptide is added to the composition, and the content of the eel peptide is replaced by hydroxyapatite. Other methods and parameters are the same as in Example 1.

[0123] Comparative Example 4

[0124] No bonito elastin peptide is added to the composition, and the content of bonito elastin peptide is replaced by hydroxyapatite. Other methods and parameters are the same as those in Example 1.

[0125] Comparative Example 5

[0126] In the preparation method of quinoa rice extract, quinoa rice is not enzymatically hydrolyzed by nattokinase, that is, it is enzymatically hydrolyzed by ficin and then directly filtered, ultrafiltered, and freeze-dried to obtain quinoa rice extract. Other methods and parameters are the same as those in Example 1.

[0127] Comparative Example 6

[0128] The composition does not contain quinoa rice extract, and the content of the quinoa rice extract is replaced by hydroxyapatite. Other methods and parameters are the same as those in Example 1.

[0129] Comparative Example 7

[0130] In the preparation method of the sapodilla rice extract, the sapodilla rice is not enzymatically hydrolyzed by subtilisin, that is, the sapodilla rice is enzymatically hydrolyzed by papain and then directly filtered, ultrafiltered, and freeze-dried to obtain the sapodilla rice extract. Other methods and parameters are the same as those in Example 1.

[0131] Comparative Example 8

[0132] No sapodilla extract is added to the composition, and the content of the components of the sapodilla extract is replaced by hydroxyapatite. Other methods and parameters are the same as in Example 1.

[0133] Comparative Example 9

[0134] The composition does not contain eel peptide, quinoa rice extract and sapodilla rice extract, and the contents of eel peptide, quinoa rice extract and sapodilla rice extract are all replaced by hydroxyapatite. Other methods and parameters are the same as those in Example 1.

[0135] Comparative Example 10

[0136] Chitosan and peach gum are not added to the composition, and the content of chitosan and peach gum is replaced by hydroxyapatite. Other methods and parameters are the same as those in Example 1.

[0137] The compositions prepared in the above-mentioned embodiments and comparative examples were tested.

[0138] Experiment 1: Safety Experiment

[0139] 150 healthy Kunming mice (weighing 24g-28g) were selected and divided into 15 groups (10 mice in each group, half male and half female), and each group was gavaged with 3000 mg / Kg of the composition prepared in each embodiment and comparative example, once a day, and fed at an interval of 2 hours after administration. The administration was continued for 30 days, and the mental state, behavioral activities, diet, and urination of the mice were observed to be normal; after being killed, no abnormal changes were observed in the heart, liver, spleen, lung, kidney and other internal organs, and the liver tissue was taken for pathological sectioning and Masson staining, and the pathological changes of the liver tissue in each group were observed under an electron microscope, and no abnormalities were found, indicating that the extract is safe and has no toxic side effects.

[0140] Experiment 2: Gum Recession Degree Experiment

[0141] 128 patients with gingival recession who met the experimental requirements were selected, half of whom were male and half were female. Inclusion criteria: aged 35 to 48 years old, no serious systemic diseases, and no use of other drugs that affect gingival recession within 2 months. Before the experiment, a periodontal probe was used to select 6 sites for measurement at multiple tooth positions (anterior teeth, posterior teeth, left and right sides) of each patient. The distance from the cementoenamel junction to the bottom of the periodontal pocket was measured and recorded as the initial gingival recession degree.

[0142] 128 patients with gingival atrophy were divided into 16 groups (8 people in each group, half male and half female) to ensure that the patients were comparable in age, gender, degree of gingival atrophy, etc. One group was a blank control group, which was not given any medicine and lived a normal life. The remaining 15 groups were drug-treated groups, each of which corresponded to the composition prepared by each embodiment and comparative example. After brushing teeth in the morning and evening, the composition was evenly applied to the teeth and gums. After applying, eating was prohibited for 4 hours and drinking water was allowed normally. After 60 days of continuous treatment, the same site was measured again using a periodontal probe and recorded as the degree of gingival recession after the test. The test results are shown in Table 1 below.

[0143] Table 1 Gingival recession degree test results (average value)

[0144]

[0145]

[0146] It can be seen from the above results that the compositions of Examples 1 to 5 can well repair gum atrophy and promote the improvement of gums to restore to a good state. It can be seen from the results of Comparative Examples 1 to 3 that the eel peptide obtained by the specific enzymatic hydrolysis step has a good effect of repairing gum atrophy. The preparation method of the eel peptide is changed, the enzymatic hydrolysis step is omitted, and the small molecule peptide product obtained has different components and different small molecule peptide contents, which greatly reduces the effect of repairing gum atrophy. It can be seen from the results of Comparative Example 4 that the bonito elastin peptide has a good auxiliary effect in repairing gum atrophy. It can be seen from the results of Comparative Examples 5 and 6 that the quinoa rice extract obtained by the specific enzymatic hydrolysis step has a good effect of repairing gum atrophy. The preparation method of the quinoa rice extract is changed, the enzymatic hydrolysis step is omitted, and the small molecule product components obtained are different, which greatly reduces the effect of repairing gum atrophy. It can be seen from the results of Comparative Examples 7 and 8 that the sapodilla rice extract obtained by the specific enzymatic hydrolysis step has a good effect of repairing gum atrophy. The preparation method of the sapodilla rice extract is changed, the enzymatic hydrolysis step is omitted, and the small molecule product components obtained are different, which greatly reduces the effect of repairing gum atrophy. From the results of Comparative Example 9, it can be seen that the eel peptide, quinoa rice extract and saponin rice extract are used in a certain proportion to significantly improve the repair effect of the composition on gingival atrophy. From the results of Comparative Example 10, it can be seen that not adding chitosan and peach gum will reduce the adhesion and moisturizing properties of the composition on teeth and gums, and the drug absorption efficiency and time are limited, which reduces the short-term use effect and prolongs the course of treatment.

[0147] Experiment 3: Gingival crevicular fluid index detection

[0148] Experiment 3 was conducted simultaneously with Experiment 2. Gingival crevicular fluid samples were collected from patients before and after the experiment to detect the levels of inflammatory factors and growth factors.

[0149] Gingival crevicular fluid collection: Gingival crevicular fluid samples were collected from 6 teeth of each patient using a gingival crevicular fluid collector. The collected gingival crevicular fluid samples were placed in a centrifuge tube and immediately stored at low temperature for subsequent testing.

[0150] The levels of inflammatory factors (interleukin-1β, tumor necrosis factor-α) and growth factors (platelet-derived growth factor) in gingival crevicular fluid and blood samples were detected by ELISA. The changes in the levels of inflammatory factors and growth factors in the experimental group and the control group before and after treatment were compared. The test results are shown in Table 2 below.

[0151] Table 2 Gingival crevicular fluid index test results (average value)

[0152]

[0153] From the above results, it can be seen that the compositions of Examples 1 to 5 can well repair gingival atrophy, improve the metabolic capacity of gingival tissue, reduce the interleukin-1β content and tumor necrosis factor-α content in gingival crevicular fluid, start the repair mechanism, and increase the content of platelet-derived growth factor. From the results of Comparative Examples 1 to 3, it can be seen that the eel peptide obtained by the specific enzymatic hydrolysis step has a good effect on repairing gingival atrophy, improving the metabolic capacity of gingival tissue, promoting the initiation of the repair mechanism, reducing inflammatory factors, and increasing growth factors; by changing the preparation method of the eel peptide and omitting the enzymatic hydrolysis step, the small molecule peptide product obtained has different components and different small molecule peptide contents, and the effect of repairing gingival atrophy is greatly reduced. From the results of Comparative Example 4, it can be seen that the bonito elastin peptide has a good auxiliary effect in repairing gingival atrophy, promoting the reduction of inflammatory factors, and increasing growth factors. It can be seen from the results of Comparative Examples 5 and 6 that the quinoa rice extract obtained by the specific enzymatic hydrolysis step has a good effect of repairing gingival atrophy, improving the metabolic capacity of gingival tissue, starting the repair mechanism, reducing inflammatory factors, and improving growth factors; the preparation method of quinoa rice extract is changed, the enzymatic hydrolysis step is omitted, and the small molecule product components obtained are different, and the effect of repairing gingival atrophy is greatly reduced. It can be seen from the results of Comparative Examples 7 and 8 that the sapodilla rice extract obtained by the specific enzymatic hydrolysis step has a good effect of repairing gingival atrophy, improving the metabolic capacity of gingival tissue, promoting the initiation of the repair mechanism, reducing inflammatory factors, and improving growth factors; the preparation method of sapodilla rice extract is changed, the enzymatic hydrolysis step is omitted, and the small molecule product components obtained are different, and the effect of repairing gingival atrophy is greatly reduced. It can be seen from the results of Comparative Example 9 that the eel peptide, quinoa rice extract and sapodilla rice extract are used in a certain proportion to greatly improve the repair effect of the composition on gingival atrophy. From the results of Comparative Example 10, it can be seen that not adding chitosan and peach gum will reduce the adhesion and moisturizing properties of the composition on teeth and gums, the drug absorption efficiency and time are limited, the efficacy is reduced, and the effect of reducing inflammatory factors and increasing growth factors in the short term is not obvious.

Claims

1. A pharmaceutical composition for preventing and treating gingival atrophy, characterized in that: The composition comprises the following raw materials in parts by weight: 0.1 to 0.5 parts of yellow eel peptide, 0.05 to 0.1 parts of bonito elastin peptide, 1 to 4 parts of quinoa rice extract, 3 to 8 parts of sapodilla rice extract, 0.5 to 3 parts of calendula extract, 0.5 to 3 parts of chamomile extract, 0.1 to 0.5 parts of chitosan, 0.2 to 0.5 parts of peach gum, 1 to 5 parts of hydroxyapatite, 0.05 to 0.08 parts of guar gum, 0.1 to 0.3 parts of soybean lecithin, and the balance is water. The water content of the material is 10% to 25%; the eel peptide is a product below 3 kDa obtained by hydrolyzing eel skin protein with trypsin and collagenase IV, and then hydrolyzing it with nattokinase and chymotrypsin; the bonito elastin peptide is a product below 3 kDa obtained by hydrolyzing bonito heart arterial bulb with porcine pancreatic elastase; the quinoa rice extract is a product below 3 kDa obtained by hydrolyzing quinoa rice with amylase, pectinase and hemicellulase, and then hydrolyzing it with ficin and nattokinase step by step; The sapodilla rice extract is obtained by hydrolyzing sapodilla rice with amylase, pectinase and hemicellulase, and then hydrolyzing it step by step with papain and subtilisin, to obtain a product with a size of less than 3 kDa.

2. A pharmaceutical composition for preventing and treating gingival atrophy according to claim 1, characterized in that: The preparation method of the yellow eel peptide comprises the following steps: taking yellow eel skin, washing it, soaking it in warm water, freezing it to remove acid, thawing it, washing it with water, draining it, and crushing it into a slurry to obtain fish skin slurry; adding a saline solution with a mass of 10 to 12 times the mass of the fish skin slurry to the fish skin slurry, stirring and soaking it, centrifuging it, collecting the supernatant, adding an ammonium sulfate aqueous solution to the supernatant, continuously stirring it until the protein is completely precipitated, centrifuging it, collecting the precipitate, and obtaining yellow eel skin protein; dispersing the yellow eel skin protein in water with a mass of 10 to 12 times the mass of the yellow eel skin protein, adjusting the pH to 7.4 to 7.8, adding trypsin and collagenase IV, and performing enzymolysis at 36° C. to 38° C. for 50 min to 90 mi n, heat up to inactivate enzyme, after cooling to 36℃~38℃, adjust pH to 8.0~8.5, add nattokinase and chymotrypsin, hydrolyze at 36℃~38℃ for 50min~70min, heat up to inactivate enzyme, after cooling to room temperature, filter with mesh, take filtrate, filter with 3kDa ultrafiltration membrane, take ultrafiltrate below 3kDa, freeze-dry, and obtain yellow eel peptide.

3. A pharmaceutical composition for preventing and treating gingival atrophy according to claim 2, characterized in that: The temperature of the warm water immersion is 40°C to 45°C, and the time of the warm water immersion is 30min to 40min; the temperature of the frozen acid removal is -15°C to -25°C, and the time of the frozen acid removal is 2h to 3h; the concentration of the saline solution is 0.1mol / L to 0.2mol / L; the rotation speed of the stirring immersion is 10r / min to 30r / min, and the stirring immersion time is 3h to 4h; the concentration of the ammonium sulfate aqueous solution is 4.0mol / L to 5.0mol / L; the rotation speed of the continuous stirring is 10r / min to 30r / min; the rotation speed of the centrifugal separation is 4000r / min to 6000r / min, and the time of the centrifugal separation is 10min to 30min; the added amounts of the trypsin and collagenase IV are both 0.5% to 1.5% of the mass of the eel skin protein; the added amounts of the nattokinase and chymotrypsin are both 0.8% to 2% of the mass of the eel skin protein.

4. A pharmaceutical composition for preventing and treating gingival atrophy according to claim 1, characterized in that: The preparation method of the bonito elastin peptide comprises the following steps: taking bonito heart artery bulbs, washing them, and crushing them into slurry, adding water 10 to 12 times the mass of the slurry into the slurry, adjusting the pH value to 7.8 to 8.5, adding 0.5 to 2% of porcine pancreatic elastase based on the mass of the bonito heart artery bulbs, performing enzymolysis at 25 to 28° C. for 2 to 3 hours, heating up to inactivate the enzyme, cooling to room temperature, filtering with a sieve, taking the filtrate, filtering with a 3 kDa ultrafiltration membrane, taking the ultrafiltrate below 3 kDa, and freeze-drying to obtain the bonito elastin peptide.

5. The pharmaceutical composition for preventing and treating gingival atrophy according to claim 1, characterized in that: The preparation method of the quinoa rice extract comprises the following steps: taking quinoa rice, adding water of 4 to 6 times the mass of quinoa rice, soaking for 40 to 60 minutes, filtering to remove water, crushing, then adding water of 8 to 12 times the mass of quinoa rice, adjusting the pH value to 4.0 to 5.0, adding 0.5% to 2% of amylase, pectinase and hemicellulase by mass of quinoa rice, performing enzymolysis at 37° C. to 45° C. for 2 hours to 4 hours, adjusting the pH value to 5.5 to 6.0, adding 0.5% to 2% of amylase, pectinase and hemicellulase by mass of quinoa rice, 1.5% of ficin is enzymatically hydrolyzed at 60-65°C for 60-90 minutes, the temperature is raised to inactivate the enzyme, the pH is adjusted to 8.0-8.5 after the temperature is lowered to 36-38°C, 0.5-1.5% of nattokinase by weight of quinoa rice is added, the temperature is enzymatically hydrolyzed at 36-38°C for 60-90 minutes, the temperature is raised to inactivate the enzyme, the temperature is lowered to room temperature, sieve is used for filtering, the filtrate is taken, a 3 kDa ultrafiltration membrane is used for filtering, the ultrafiltrate below 3 kDa is taken, and freeze-drying is performed to obtain a quinoa rice extract.

6. The pharmaceutical composition for preventing and treating gingival atrophy according to claim 1, characterized in that: The preparation method of the sapodilla rice extract comprises the following steps: adding sapodilla rice to water of 4 to 6 times the mass of sapodilla rice, soaking for 40 to 60 minutes, filtering to remove water, crushing, then adding water of 8 to 12 times the mass of sapodilla rice, adjusting the pH to 4.0 to 5.0, adding 0.5% to 2% of amylase, pectinase and hemicellulase by mass of sapodilla rice, performing enzymolysis at 37° C. to 45° C. for 2 hours to 4 hours, adjusting the pH to 5.7 to 6.5, adding 0.5% to 1. 5% papain is enzymolyzed at 55-60°C for 60-90 min, the temperature is raised to inactivate the enzyme, the pH is adjusted to 8.0-9.0 after the temperature is lowered to 50-60°C, 0.5%-1.5% of subtilisin by weight of sapodilla rice is added, the temperature is raised to inactivate the enzyme, the temperature is lowered to room temperature, sieve is used for filtering, filtrate is taken, 3 kDa ultrafiltration membrane is used for filtering, ultrafiltrate below 3 kDa is taken, and freeze-drying is performed to obtain sapodilla rice extract.

7. A pharmaceutical composition for preventing and treating gingival atrophy according to claim 2, 4, 5 or 6, characterized in that: The temperature of the heating enzyme inactivation is 80°C to 90°C, and the time of the heating enzyme inactivation is 15min to 25min; the mesh number of the sieve is 250 meshes to 325 meshes.

8. The pharmaceutical composition for preventing and treating gingival atrophy according to claim 1, characterized in that: The calendula extract is an ethanol percolation extract. The preparation method of the calendula extract comprises the following steps: taking the flowers of the calendula, crushing them to obtain a crushed product, using ethanol 6 to 8 times the weight of the flowers to percolate the crushed product to obtain a percolate, concentrating and drying the percolate to obtain the calendula extract; The chamomile extract is an ethanol percolation extract. The preparation method of the chamomile extract comprises the following steps: taking chamomile flowers, crushing to obtain a crushed product, using ethanol 6 to 8 times the mass of the flowers to percolate the crushed product to obtain a percolate, concentrating, and drying to obtain the chamomile extract.

9. A method for preparing a pharmaceutical composition for preventing and treating gum atrophy, for preparing the pharmaceutical composition for preventing and treating gum atrophy according to claim 1, characterized in that: The preparation method comprises the following steps: S1: Mix eel peptide, bonito elastin peptide, quinoa rice extract, sapodilla rice extract, calendula extract and chamomile extract according to weight percentage to obtain a mixture A; S2: adding water of twice the mass of mixture A to mixture A, mixing and dispersing the mixture evenly to obtain a mixture B; S3: crushing peach gum according to mass fraction, adding peach gum into water at a mass ratio of peach gum: water = 1: (3-5) to soak for 6h-10h to obtain a foamed material C; S4: taking guar gum according to mass fraction, dissolving the guar gum in water at a mass ratio of guar gum: water = 1: (100-120) to obtain a glue solution D; S5: Mix the mixed solution B, foaming material C, glue solution D and soybean lecithin in proportions by mass, then add chitosan and hydroxyapatite, mix well, and concentrate under negative pressure below 55° C. to a water content of 10% to 25% to obtain a composition.

10. Use of the pharmaceutical composition for preventing and treating gingival atrophy according to claim 1, characterized in that: The composition is used for preparing a medicament for coating the gum area for preventing and treating gum atrophy.

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