Pharmaceutical composition as well as preparation method and application thereof

Abstract

The invention provides a pharmaceutical composition as well as a preparation method and application thereof. The pharmaceutical composition comprises the following raw materials in parts by mass: 2-6 parts of radix paeoniae alba, 1-4 parts of ginseng, 1-4 parts of mint and 0.3-3 parts of liquorice. The pharmaceutical composition provided by the invention adopts the radix paeoniae alba, the ginseng, the mint and the liquorice, the mint and the liquorice are medicinal and edible traditional Chinese medicines, and the radix paeoniae alba and the ginseng can be widely applied in a health food raw material list. The pharmaceutical composition is a pure traditional Chinese medicine preparation, has significant antibacterial and anti-inflammatory effects, can effectively inhibit gingival bleeding, reduce dental plaque, reduce gingival sulcus bleeding index, gingival index and dental plaque index, and significantly reduce probe depth, and has a good curative effect. The composition has a remarkable treatment effect on gingivitis and periodontitis and is free of toxic and side effects.

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical technology, and specifically relates to a pharmaceutical composition, a preparation method thereof, and an application thereof. Background Art

[0002] Gingivitis and periodontitis are common oral diseases, bringing great pain and inconvenience to patients. At present, clinically, mainly antibiotics and other drugs are used for treatment, but long-term use of antibiotics is likely to produce drug resistance and may have some side effects. Therefore, finding a safe and effective method for preventing and treating gingivitis and periodontitis has important practical significance.

[0003] Compared with Western medicines, most traditional Chinese medicines are derived from natural plants and have relatively few side effects. When preventing and treating gingivitis and periodontitis, traditional Chinese medicines generally do not cause problems such as drug resistance and dysbacteriosis like Western medicines such as antibiotics. Long-term use of traditional Chinese medicines for treatment also causes less damage to important organs such as the liver and kidneys of the human body and is more suitable for long-term use by patients. Patent application CN109731051A discloses an extract of Fujian narcissus tea, a preparation method thereof, and an application thereof in inhibiting periodontitis pathogenic bacteria. Patent application CN115887589A discloses a pharmaceutical composition for treating chronic periodontitis, a preparation method thereof, and uses, which is composed of Smilax glabra, Polygonatum sibiricum, Taraxacum mongolicum, Portulaca oleracea, Agrimonia pilosa, Plantago asiatica, Coix lacryma-jobi, Ligusticum wallichii, Cyathula officinalis, Sepia seu cuttlebone, Fructus mume, and Glycyrrhiza uralensis. Patent application CN114191484A discloses a Miao medicine solid tooth formula for treating periodontitis and a preparation method thereof, which is composed of Qingyan, Gypsum Fibrosum, Dipsacus asperoides, Sargentodoxa cuneata, Begonia cathayana, Rhododendron molle, Blumea balsamifera, Selaginella tamariscina, and Paederia scandens. The above patent applications disclose different plant extracts, traditional Chinese medicine formulas, preparation methods, and taking methods for treating periodontitis. However, at present, the traditional Chinese medicine compositions of these patent applications for treating periodontitis have relatively many medicinal flavors and are not suitable for long-term use, and contain drugs that are not both medicine and food or raw materials for health foods, and there may be safety problems. Summary of the Invention

[0004] In order to solve the above technical problems, the present invention provides a composition for preventing and treating gingivitis and periodontitis and a preparation method thereof. The white peony root, ginseng, mint, and licorice used in the pharmaceutical composition provided by the present invention are all on the list of traditional Chinese medicines that are both medicine and food or raw materials for health foods. The pharmaceutical composition is a pure traditional Chinese medicine preparation, which has the functions of antibacterial and anti-inflammatory, inhibiting gingival bleeding, reducing dental plaque, reducing the gingival sulcus bleeding index, reducing the gingival index, reducing the dental plaque index, and reducing the probing depth, has a significant effect on gingivitis and periodontitis, and has no toxic side effects.

[0005] In a first aspect, the pharmaceutical composition provided by the present invention comprises raw materials in the following parts by mass: 2 - 6 parts of white peony root, 0.5 - 4 parts of ginseng, 0.5 - 4 parts of mint, and 0.2 - 3 parts of licorice root. The traditional Chinese medicine composition provided by the present invention has significantly reduced the number of medicinal flavors, is more suitable for long-term use, and the raw materials used are all on the list of traditional Chinese medicines with both medicinal and food uses or raw materials for health foods, greatly solving the safety problems existing in the existing methods. Moreover, this composition uses white peony root as the core ingredient. By utilizing the interaction of a certain amount of white peony root, ginseng, mint, and licorice root, while obtaining a pure traditional Chinese medicine composition with the characteristics of both medicinal and food uses, it also has multiple effects such as antibacterial and anti-inflammatory, reducing gingival bleeding, and lowering the plaque index, and has no toxic side effects. It can be better used for the treatment of gingivitis and periodontitis, and has good application prospects.

[0006] Preferably, the pharmaceutical composition comprises raw materials in the following parts by mass: 2 - 5 parts of Bozhou white peony root, 0.8 - 3 parts of ginseng, 0.8 - 3 parts of mint, and 0.3 - 2 parts of licorice root. In the present invention, under the preferred dosages, Bozhou white peony root, ginseng, mint, and licorice root can better interact with each other, which is beneficial to further improving the therapeutic effect on gingivitis and periodontitis.

[0007] More preferably, the pharmaceutical composition comprises raw materials in the following parts by mass: 2.5 - 3.5 parts of Bozhou white peony root, 1 - 1.5 parts of ginseng, 1 - 1.5 parts of mint, and 0.5 - 1 part of licorice root. For example, Bozhou white peony root can be 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5 parts, etc. and any numerical values and ranges therebetween, ginseng can be 1, 1.1, 1.2, 1.3, 1.4, 1.5 parts, etc. and any numerical values and ranges therebetween, mint can be 1, 1.1, 1.2, 1.3, 1.4, 1.5 parts, etc. and any numerical values and ranges therebetween, and licorice root can be 0.5, 0.6, 0.7, 0.8, 0.9, 1 part, etc. and any numerical values and ranges therebetween.

[0008] Even more preferably, the raw materials of the pharmaceutical composition are 3 parts of Bozhou white peony root, 1 - 1.2 parts of ginseng, 1 - 1.2 parts of mint, 0.5 - 0.6 parts of licorice root, and 55 - 100 parts of water; preferably, the dosage of water is 10 - 18 times the total mass of Bozhou white peony root, ginseng, mint, and licorice root, such as 11, 12, 13, 14, 15, 16, 17 times, etc. and any numerical values and ranges therebetween. The present invention discovers that using the above raw materials under further preferred dosages can better exert the synergistic effect of Bozhou white peony root, ginseng, mint, and licorice root, thereby significantly improving the therapeutic effects such as oral antibacterial and anti-inflammatory, inhibiting gingival bleeding, reducing dental plaque, lowering the sulcus bleeding index, lowering the gingival index, lowering the plaque index, and lowering the probing depth. Most preferably, by mass, it is 3 parts of Bozhou white peony root, 1 part of ginseng, 1 part of mint, 0.5 part of licorice root, and 77 parts of water.

[0009] In a second aspect, the present invention provides a method for preparing the above-mentioned pharmaceutical composition, comprising: 1) Extraction: Mix Paeonia lactiflora Pall., ginseng, Mentha haplocalyx Briq., liquorice and water, decoct and filter to obtain an extract.

[0010] 2) Concentration: Concentrate the extract under reduced pressure to obtain a concentrated solution.

[0011] 3) Drying: Spray-dry the concentrated solution.

[0012] Preferably, in step 1), the number of decoctions is at least two times. For the first decoction, add 6 - 10 times the amount of water, and the decoction time is 1 - 2 h, then filter. For the second decoction, add 4 - 8 times the amount of water, and the decoction time is 0.5 - 1.5 h.

[0013] More preferably, the number of decoctions is two times. For the first decoction, add 8 ± 1 times the amount of water, and the decoction time is 1.5 ± 0.2 h, then filter. For the second decoction, add 6 ± 1 times the amount of water, and the decoction time is 1.0 ± 0.2 h.

[0014] More preferably, the decoction temperature can be set at the conventional temperature in the art, and the present invention does not limit this.

[0015] Preferably, in step 2), the temperature for concentration under reduced pressure is 55 - 75 °C, and the pressure is -0.08 - -0.04 Mpa; the relative density of the concentrated solution (at 60 °C) is 1.08 - 1.15.

[0016] Preferably, in step 3), the inlet air temperature is 160 - 190 °C, the outlet air temperature is 70 - 100 °C, the rotation speed of the atomizer is 40 - 50 HZ, and the flow rate of the infusion pump is 7 - 15 HZ.

[0017] In the present invention, under the above-mentioned preferred methods and conditions, a pharmaceutical composition with excellent effects can be better prepared.

[0018] In a third aspect, the present invention provides a medicament, which comprises the above-mentioned pharmaceutical composition or the pharmaceutical composition obtained by the preparation method of the above-mentioned pharmaceutical composition; preferably, the dosage form of the medicament is powder, ointment, tablet, pill, capsule, granule, infusion or oral liquid. In the present invention, by adding a pharmaceutically acceptable carrier and / or excipient to the dry extract powder of the above-mentioned pharmaceutical composition, any clinically acceptable dosage form can be prepared by using traditional preparation processes.

[0019] Fourthly, the present invention provides the use of the above-mentioned pharmaceutical composition, or the pharmaceutical composition obtained by the above-mentioned preparation method, or the above-mentioned medicament in the preparation of a medicament for preventing and treating gingivitis and / or periodontitis. Preferably, the use of the pharmaceutical composition or the medicament in one or more of oral antibacterial and anti-inflammatory, inhibiting gingival bleeding, reducing dental plaque, reducing the sulcus bleeding index, reducing the gingival index, reducing the dental plaque index, and reducing the probing depth.

[0020] The beneficial effects of the present invention are at least as follows: The pharmaceutical composition of the present invention is a pure traditional Chinese medicine preparation, which has the functions of antibacterial and anti-inflammatory, inhibiting gingival bleeding, reducing dental plaque, reducing the sulcus bleeding index, reducing the gingival index, reducing the dental plaque index, and reducing the probing depth, and has a significant effect on gingivitis and periodontitis, and has no toxic and side effects. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] In order to more clearly illustrate the technical solutions in the present invention or the prior art, the following will briefly introduce the drawings required for use in the description of the embodiments or the prior art. Obviously, the drawings in the following description are some embodiments of the present invention. For those of ordinary skill in the art, other drawings can be obtained based on these drawings without creative efforts.

[0022] Figure 1 It shows the gingival bleeding condition of the gingivitis rats provided in Experimental Example 1 of the present invention.

[0023] Figure 2 It shows the dental plaque staining of the gingivitis rats provided in Experimental Example 1 of the present invention.

[0024] Figure 3 It shows the gingival bleeding condition of the gingivitis rats provided in Experimental Example 1 of the present invention.

[0025] Figure 4 It shows the dental plaque condition of the gingivitis rats provided in Experimental Example 1 of the present invention.

[0026] Figure 5 It shows the gingival bleeding condition of the periodontitis rats provided in Experimental Example 2 of the present invention.

[0027] Figure 6 It shows the dental plaque staining of the periodontitis rats provided in Experimental Example 2 of the present invention.

[0028] Figure 7 It shows the gingival bleeding condition of the periodontitis rats provided in Experimental Example 2 of the present invention.

[0029] Figure 8 It shows the dental plaque condition of the periodontitis rats provided in Experimental Example 2 of the present invention. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0030] To make the objectives, technical solutions, and advantages of the present invention clearer, the technical solutions in the present invention will be clearly and completely described below. Apparently, the described embodiments are some, but not all, of the embodiments of the present invention. All other embodiments obtained by those of ordinary skill in the art based on the embodiments in the present invention without creative efforts belong to the scope of protection of the present invention.

[0031] In the ranges disclosed in the present invention, the endpoints and any values are not limited to the exact ranges or values. These ranges or values should be understood to include values close to these ranges or values. For numerical ranges, between the endpoint values of each range, between the endpoint values of each range and individual point values, and between individual point values, they can be combined with each other to obtain one or more new numerical ranges, and these numerical ranges should be regarded as specifically disclosed in the present invention.

[0032] In the embodiments of the present invention, where specific technologies or conditions are not indicated, they are carried out according to the technologies or conditions described in the literature in the field or according to the product specifications. For devices, instruments, reagents, etc. where the manufacturer is not indicated, they are all conventional products that can be obtained through regular channels. The experimental reagents and raw materials involved are all commercially available products, and the reagents are all analytical pure products.

[0033] Example 1 This example provides a pharmaceutical composition, which is composed of the following raw materials in parts by mass: 3 parts of Radix Paeoniae Alba, 1 part of Ginseng, 1 part of Mentha haplocalyx Briq., and 0.5 part of Glycyrrhiza uralensis Fisch.

[0034] This embodiment also provides a preparation method of the pharmaceutical composition, which is as follows: In the above raw materials, the Bozhou white peony root is harvested from August to October. First, wash the soil on the peony root, and then use tools such as bamboo knives, porcelain chips or glass slides to scrape off the outer skin. Put the peeled peony root into a pot and add an appropriate amount of water for boiling. The amount of water for boiling should be sufficient to cover the peony root. Bring the water to a boil over high heat, and then turn to low heat and cook for 10 minutes. Put the cooked peony root in a drying device for drying at a temperature of 60°C. Put the Bozhou white peony root medicinal material into a suitable container, add an appropriate amount of water, preferably covering the medicinal material, and soak for 3 hours for softening. Put the softened Bozhou white peony root neatly into the feed inlet of the slicing machine, adjust the slicing thickness parameter of the slicing machine to 2 mm, and slice. Put the sliced Bozhou white peony root decoction pieces into a drying device for drying at a temperature of 60°C. Remove the soil, sand, remaining rhizome, lateral roots, etc. from the dug ginseng. Soak the ginseng in an appropriate amount of clean water for 8 hours. Cut the softened ginseng into thin slices with a slicing machine, with a thickness of about 2 mm. Put the cut ginseng decoction pieces into a drying device for drying at a temperature set at 50°C. Remove the impurities such as soil, sand, weeds, etc. from the harvested whole herb of Mentha haplocalyx Briq., and then quickly rinse it with clean water. Put the rinsed Mentha haplocalyx Briq. into a drying device for drying treatment at a temperature set at 50°C. Remove the impurities such as soil, sand, weeds, etc. from the dug licorice root, and then rinse it clean by means of flowing water flushing. Use a medicine cutting machine to cut the licorice root into slices with a thickness of 2 mm. Dry the licorice slices in a drying device at a temperature set at 60°C. Weigh the decoction pieces in step 2 according to the above dosage, add 8 times the amount of water, decoct at 100°C for 1.5 hours, filter out the filtrate, then add 6 times the amount of water, decoct at 100°C for 1 hour, and filter out the filtrate. Combine the filtrates from the two decoctions, concentrate under reduced pressure at 60°C until the relative density is 1.08 (60°C), with the feed pump frequency of 15 HZ, the atomizer frequency of 50 HZ, the inlet air temperature of 180°C, the outlet air temperature of 90°C, and spray-dry the concentrated solution to obtain the pharmaceutical composition in the form of dry extract powder.

[0035] Example 2 This example uses the same pharmaceutical composition and method as in Example 1, and the only difference is that the dosage of the raw material composition is in the following combination: 2 parts of Bozhou white peony root, 0.5 part of ginseng, 0.5 part of Mentha haplocalyx Briq., and 0.2 part of licorice root.

[0036] Example 3 This example uses the same pharmaceutical composition and method as in Example 1, and the only difference is that the dosage of the raw material composition is in the following combination: 6 parts of Bozhou white peony root, 4 parts of ginseng, 4 parts of Mentha haplocalyx Briq., and 3 parts of licorice root.

[0037] Example 4 This example uses the same pharmaceutical composition and method as in Example 1, and the only difference is that the dosage of the raw material composition is in the following combination: 3 parts of Bozhou white peony root, 1 part of ginseng, 1 part of Mentha haplocalyx Briq., and 0.5 part of licorice root.

[0038] Experimental Example 1 1. Materials Drug Lipopolysaccharide (LPS, provided by Sigma - Aldrich, USA) Animals Sprague - Dawley (SD) rats were provided by Beijing Vital River Laboratory Animal Technology Co., Ltd. 2. Methods and Results 2.1 Preparation of Gingivitis Model Forty 8 - week - old SD rats, weighing about 200 g, were evenly divided into 5 groups: normal control group; gingivitis model group; low - dose administration (30 mg / kg) group; high - dose administration (100 mg / kg) group; and minocycline hydrochloride ointment group. Rats in groups other than the normal control group were established with a gingivitis model by lipopolysaccharide injection combined with 10% white sugar drinking water. A 20% urethane solution was prepared and the rats were intraperitoneally anesthetized at a dose of 4 mL / kg. A micro - syringe was used to accurately aspirate 10 μL of LPS - PG stock solution with a concentration of 1 mg / mL and slowly injected into the palatal gingiva between the first and second maxillary molars on both sides (i.e., the right and left sides) of the rats' oral cavity, once every 48 h, for a total of 4 times. During the injection process, it was necessary to ensure that the syringe needle accurately penetrated the target position and the operation was gentle to avoid damaging the surrounding tissues and causing strong stress reactions in the rats. After each lipopolysaccharide injection, the rats' drinking water was immediately changed to 10% white sugar aqueous solution and continuously provided until the next injection. During this period, the water intake and general condition of the rats were closely observed to ensure that the rats could normally ingest the white sugar water and had no obvious discomfort or abnormal behavior. At the same time, the breeding environment was kept clean and hygienic, and the cage bedding was regularly changed to prevent interference with the experimental results due to environmental factors.

[0039] The rats' gums were observed. If there were gingival margin swelling, brightness, softness, and easy bleeding when touched, it was regarded as successful construction of the gingivitis rat model. Compared with the normal group, the model group had obvious gingival bleeding ( Figure 1 ), and the degree of dental plaque staining was also deeper ( Figure 2 ), indicating that there were more dental plaques in the model group.

[0040] 2.2 Drug Administration In the administration groups, the periodontal tissues of the rats were smeared with drug powder (Example 1) by cotton swabs every day according to the rats' body weight. The low - dose group was at 30 mg / kg, the high - dose group was at 100 mg / kg, and each smear was maintained for about 1 min, twice a day, for 7 consecutive days. The model was smeared with 0.9% sodium chloride solution, and the positive control group was smeared with minocycline hydrochloride ointment.

[0041] 2.3 Detection of Gingival Index Using a blunt periodontal probe, examine the mesial buccal papilla, middle buccal margin, distal buccal papilla, and lingual gingival margin of the gingiva around the left maxillary M2 molar in rats, and take the average according to the scoring criteria. Examine the mesial buccal papilla, middle buccal margin, distal buccal papilla, and lingual gingival margin of the gingiva around the left maxillary M2 molar, score them respectively according to the following scoring criteria, and take the average. Scoring criteria: 0 = healthy gingiva, no redness, swelling, or bleeding; 1 = mild inflammation, no bleeding when gently probed with a probe. The gingiva has color changes and mild edema; 2 = moderate inflammation, bleeding when gently probed with a probe, gingiva is red and swollen; 3 = severe inflammation, tendency of spontaneous bleeding, gingiva is significantly red and swollen or has ulcers.

[0042] The results are shown in Table 1. The gingival index of the normal group of rats was 0, the gingival index of the gingivitis model group was 2.563 ± 0.2588, and the gingival indices of the low-dose administration group and the high-dose administration group were 1.906 ± 0.2969 and 1.375 ± 0.2315 respectively, all of which were significantly different from the model group ( P <0.0001). The gingival index of the minocycline hydrochloride ointment group was 0.9286 ± 0.3134, showing a good effect.

[0043] Table 1 Gingival Index of Rats with Gingivitis

[0044] **** P <0.0001, compared with the model group 2.4 Detection of Gingival Bleeding Gently insert the periodontal probe below the gingival margin and slide it, observe the degree of gingival bleeding, and score according to the scoring criteria. When examining, probe the mesial, palatal, and distal sites of the left maxillary M2 in rats, and take the average. Scoring criteria: 0 = no bleeding when gently probed with a periodontal probe, the gingiva looks healthy without redness or swelling; 1 = no bleeding when gently probed with a periodontal probe, the gingiva is slightly red and swollen, with mild inflammation; 2 = punctate bleeding of the gingiva when gently probed with a periodontal probe, the appearance of the gingiva has color changes and edema; 3 = bleeding of the gingiva extends along the gingival margin when gently probed with a periodontal probe, the gingiva has obvious swelling; 4 = bleeding of the gingiva overflows the gingival margin when gently probed with a periodontal probe, showing severe inflammation, the gingiva has obvious swelling; 5 = spontaneous bleeding, the gingiva has color changes and obvious swelling, and sometimes has ulcers.

[0045] The results are as Figure 3As shown, rats in the gingivitis model group had obvious symptoms of gingival bleeding. However, in both the low-dose and high-dose groups of the drug-administered group, the symptoms of gingival bleeding in rats were significantly improved, and the effect was similar to that of the minocycline hydrochloride ointment group, indicating that the drug has an obvious effect on improving gingival bleeding in gingivitis. The results of the sulcus bleeding index are shown in Table 2. The sulcus bleeding index of rats in the normal group was 0, that of rats in the gingivitis model group was 3.833 ± 0.6172, and those of the low-dose and high-dose drug-administered groups were 2.292 ± 0.7440 and 1.625 ± 0.2782 respectively, all of which were significantly different from the model group ( P <0.0001), and the sulcus bleeding index of the minocycline hydrochloride ointment group was 1.095 ± 0.2520, showing a good effect.

[0046] Table 2 Sulcus bleeding index of rats with gingivitis

[0047] **** P <0.0001, compared with the model group 2.5 Dental plaque detection First, dip a cotton ball in pure water and gently wipe the rat's teeth to clean the oral cavity. Then, dip a cotton ball in basic fuchsin staining agent and wipe the bilateral maxillary first molars. After staining, use a dry cotton ball to absorb the excess staining solution on the tooth surface. After staining, use a dental probe to gently scratch the gingival sulcus, gingival margin and adjacent tooth surfaces of the bilateral maxillary first molars of the rats to check whether plaque can be scraped out. Carefully observe all surfaces of the rat's teeth, including the labial (buccal) surface, lingual surface, mesial surface and distal surface, and use visual inspection combined with gentle scratching with a probe to evaluate the plaque situation. Score 0: On all tooth surfaces of the rat's teeth, including the gingival margin, there is no visible plaque to the naked eye, and no plaque can be felt when gently scratching the tooth surface with a probe. This indicates that there is very little plaque in the rat's oral cavity. Score 1: On some tooth surfaces of the teeth (such as the labial surface or lingual surface), a thin plaque layer can be found when gently scratching with a probe, but it is difficult to see the plaque directly with the naked eye. It shows that a small amount of plaque begins to attach to the tooth surface. Score 2: Visible plaque can be seen on multiple tooth surfaces of the teeth (such as the labial surface and mesial surface) with the naked eye, and the coverage area of the plaque is relatively large, but the thickness is not particularly thick. Score 3: A large amount of plaque accumulates on most tooth surfaces of the rat's teeth, including around the gingival margin, the thickness of the plaque is obvious, and even some plaque may begin to mineralize to form the initial state of dental calculus.

[0048] The results are as Figure 4As shown, obvious plaque staining was observed on the teeth of rats in the gingivitis model group. Compared with the model group, the degree of dental plaque staining in the low- and high-dose drug administration groups was significantly reduced. The effect of the high-dose group was better than that of the low-dose group, and the color and texture were close to those of the normal group. The effect of the minocycline hydrochloride ointment group was relatively significant. The results of the dental plaque index are shown in Table 2. The dental plaque index of rats in the normal group was 0, that of rats in the gingivitis model group was 2.500 ± 0.2673, and that of the low-dose drug administration group was 2.094 ± 0.1294, which was significantly lower than that of the model group ( P <0.05), the dental plaque index of the high-dose drug administration group was 1.500 ± 0.3273, showing a significant difference from the model group ( P <0.0001), and the dental plaque index of the minocycline hydrochloride ointment group was 1.000 ± 0.3819, showing a good effect.

[0049] Table 3 Dental Plaque Index of Rats with Gingivitis

[0050] * P <0.05, **** P <0.0001, compared with the model group Experimental Example 2 1. Materials 1.1 Drugs Lipopolysaccharide (provided by Sigma-Aldrich, USA) 1.2 Animals Sprague-Dawley rats were provided by Beijing Vital River Laboratory Animal Technology Co., Ltd. 2. Methods and Results 2.1 Preparation of Periodontitis Model Forty 8-week-old Sprague-Dawley rats, weighing about 200 g, were evenly divided into 5 groups: normal control group; gingivitis model group; low-dose administration (30 mg / kg) group; high-dose administration (100 mg / kg) group; and minocycline hydrochloride ointment group. Rats in groups other than the normal control group were established with a gingivitis model by lipopolysaccharide injection combined with 10% sucrose water for drinking. A 20% urethane solution was prepared and the rats were anesthetized intraperitoneally at a dose of 4 mL / kg. Using a micro syringe, 10 μL of LPS-PG stock solution with a concentration of 1 mg / mL was precisely aspirated and slowly injected into the palatal gingiva between the first and second maxillary molars on both sides (i.e., the right and left sides) of the rats' oral cavity, once every 48 h, for a total of 8 injections. During the injection process, it was necessary to ensure that the syringe needle accurately penetrated the target position and the operation was gentle to avoid damaging the surrounding tissues and causing strong stress responses in the rats. Immediately after each lipopolysaccharide injection, the rats' drinking water was replaced with a 10% sucrose aqueous solution and continuously provided until the next injection. During this period, the water intake and general condition of the rats were closely observed to ensure that the rats could normally ingest the sucrose water without obvious discomfort or abnormal behavior. At the same time, the breeding environment was kept clean and hygienic, and the cage bedding was regularly changed, etc., to prevent interference with the experimental results due to environmental factors.

[0051] As Figure 5 shown, compared with the normal group, the periodontitis model rats had obvious gingival bleeding, the gingiva showed dark red, mild edema, shiny surface, soft texture, and bleeding, and the probing depth of the periodontium was greater than 1 mm. In addition, compared with the normal group, the plaque staining degree of the periodontitis model rats was deeper, indicating that the periodontitis model was successfully established in the rats.

[0052] 2.2 Drug administration In the drug administration groups, the periodontal tissues of the rats were smeared with the drug powder (Example 1) using cotton swabs every day according to the rats' body weights. The low-dose group was at 30 mg / kg, and the high-dose group was at 100 mg / kg. Each smear was maintained for about 1 min, twice a day, for 7 consecutive days. The model group was smeared with 0.9% sodium chloride solution, and the positive control group was smeared with minocycline hydrochloride ointment.

[0053] 2.3 Gingival index detection Using a blunt periodontal probe, examine the mesial buccal papilla, middle buccal margin, distal buccal papilla, and lingual gingival margin of the gingiva around the left maxillary M2 molar in rats, and score according to the scoring criteria and take the average. Examine the mesial buccal papilla, middle buccal margin, distal buccal papilla, and lingual gingival margin of the gingiva around the left maxillary M2 molar, score according to the following scoring criteria respectively, and take the average. Scoring criteria: 0 = healthy gingiva, no redness, swelling, or bleeding; 1 = mild inflammation, no bleeding when gently probed with a probe. The gingiva has color changes and mild edema; 2 = moderate inflammation, bleeding when gently probed with a probe, gingiva is red and swollen; 3 = severe inflammation, tendency of spontaneous bleeding, gingiva is significantly red and swollen or has ulcers.

[0054] The results are shown in Table 4. The gingival index of rats in the normal group was 0, the gingival index of rats in the periodontitis model group was 2.250 ± 0.4629, and the gingival index of rats in the low-dose administration group was 1.905 ± 0.2520. Although there was a decrease, there was no significant difference compared with the model group. The gingival index of rats in the high-dose administration group was 0.8571 ± 0.4658, and there was a significant difference compared with the model group ( P <0.0001), and the gingival index of the minocycline hydrochloride ointment group was 0.5238 ± 0.3563, showing a good effect.

[0055] Table 4 Gingival Index of Periodontitis Rats

[0056] **** P <0.0001, compared with the model group 2.4 Detection of Gingival Bleeding Gently insert a periodontal probe below the gingival margin and slide it to observe the degree of gingival bleeding, and score according to the scoring criteria. When examining, probe three sites: mesial, palatal, and distal of the left maxillary M2 in rats, and take the average. Scoring criteria: 0 = no bleeding when gently probed with a periodontal probe, healthy gingiva appearance without redness and swelling; 1 = no bleeding when gently probed with a periodontal probe, slightly red and swollen gingiva, mild inflammation; 2 = punctate bleeding of the gingiva when gently probed with a periodontal probe, color changes and edema in the gingiva appearance; 3 = bleeding of the gingiva extends along the gingival margin when gently probed with a periodontal probe, obvious swelling of the gingiva; 4 = bleeding of the gingiva overflows the gingival margin when gently probed with a periodontal probe, severe inflammation, obvious swelling of the gingiva; 5 = spontaneous bleeding, color changes and obvious swelling of the gingiva, sometimes with ulcers.

[0057] The results are as Figure 7As shown, the gums of the rats in the periodontitis model group were swollen and red, with obvious bleeding. Compared with the model group, the swelling and redness of the gums in the low- and high-dose drug administration groups were significantly reduced, and the color and texture were close to those of the normal group. The effect of the high-dose group was better than that of the low-dose group. Minocycline hydrochloride ointment had a significant effect on improving bleeding and swelling, indicating that the drug had an obvious effect on improving gingival bleeding in periodontitis. The results of the sulcus bleeding index are shown in Table 5. The sulcus bleeding index of the rats in the normal group was 0, that of the rats in the periodontitis model group was 4.042 ± 0.6026, and those of the low- and high-dose drug administration groups were 2.714 ± 0.2999 and 1.762 ± 0.3171, respectively, all of which were significantly different from the model group ( P <0.0001). The sulcus bleeding index of the minocycline hydrochloride ointment group was 0.6190 ± 0.3563, showing a good effect.

[0058] Table 5 Sulcus bleeding index of rats with periodontitis

[0059] **** P <0.0001, compared with the model group 2.5 Dental plaque detection First, dip a cotton ball in pure water and gently wipe the rat's teeth to clean the oral cavity. Then, dip a cotton ball in basic fuchsin staining agent and wipe the bilateral maxillary first molars. After staining, use a dry cotton ball to absorb the excess staining solution on the tooth surface. After staining, use a dental probe to gently scrape across the gingival sulcus, gingival margin, and adjacent tooth surfaces of the bilateral maxillary first molars of the rats to check if plaque can be scraped off. Carefully observe all surfaces of the rat's teeth, including the labial (buccal) surface, lingual surface, mesial surface, and distal surface, and use visual inspection combined with gentle scraping with a probe to evaluate the plaque situation. Score 0: On all tooth surfaces of the rat's teeth, including the gingival margin, there is no visible plaque to the naked eye, and no plaque can be felt when gently scraping the tooth surface with a probe. This indicates that there is very little plaque in the rat's oral cavity. Score 1: On some tooth surfaces of the teeth (such as the labial surface or lingual surface), a thin plaque layer can be found when gently scraping with a probe, but it is difficult to see the plaque directly with the naked eye. This shows that a small amount of plaque has begun to attach to the tooth surface. Score 2: Visible plaque can be seen on multiple tooth surfaces of the teeth (such as the labial surface and mesial surface) with the naked eye, and the coverage area of the plaque is relatively large, but the thickness is not particularly thick. Score 3: A large amount of plaque accumulates on most tooth surfaces of the rat's teeth, including around the gingival margin. The thickness of the plaque is obvious, and even some plaque may begin to mineralize to form the initial state of dental calculus.

[0060] The results are as Figure 8As shown, the teeth of the rats in the periodontitis model group had obvious plaque staining; compared with the model group, the degree of plaque staining on the teeth of the rats in the low- and high-dose drug administration groups was significantly reduced, and the effect of the high-dose group was better than that of the low-dose group, and the effect of the minocycline hydrochloride ointment group was more significant. The results of the plaque index are shown in Table 6. The plaque index of the rats in the normal group was 0, the plaque index of the rats in the periodontitis model group was 2.219 ± 0.3644, and the plaque index of the rats in the low-dose drug administration group was 1.857 ± 0.5926. Although the value decreased, there was no significant difference compared with the model group. The plaque index of the rats in the high-dose drug administration group was 1.071 ± 0.2782, showing a significant difference compared with the model group ( P <0.0001), and the plaque index of the minocycline hydrochloride ointment group was 0.4286 ± 0.2782, showing a good effect. The results indicate that the drug has a good effect in inhibiting periodontitis dental plaque.

[0061] Table 6 Plaque Index of Rats with Periodontitis

[0062] **** P <0.0001, compared with the model group 2.6 Probing Depth Detection The probing depth is an important periodontal clinical index. The depth of the periodontal pocket is measured using a periodontal probe, and the distance from the gingival margin to the bottom of the gingival sulcus is the probing depth.

[0063] Examination method: Use a periodontal probe to examine the depth of the periodontal pocket of the rats parallel to the long axis of the tooth, and explore the depths of the mesial, palatal, and distal sites of the left maxillary M2 molar of the anesthetized rats, and take the average value.

[0064] The results of the probing depth of the rats with periodontitis are shown in Table 7. The probing depth of the rats in the normal group was 0 mm, the probing depth of the rats in the periodontitis model group was 1.583 ± 0.2357 mm, and the probing depth of the rats in the low-dose drug administration group was 1.143 ± 0.3253 mm, showing a significant decrease compared with the model group ( P <0.05), the probing depth of the rats in the high-dose drug administration group was 0.7619 ± 0.4179, showing a significant difference compared with the model group ( P <0.0001), and the probing depth of the rats in the minocycline hydrochloride ointment group was 0.4286 ± 0.370 mm, showing a good effect.

[0065] Table 7 Probing Depth of Rats with Periodontitis

[0066] * P <0.05, **** P <0.0001, compared with the model group The methods of Experimental Example 1 were used to test Examples 2 - 4, and the test results are shown in Tables 8 to 10.

[0067] Example 2 was tested. In the gingivitis rat model, there were no significant differences in the gingival index and sulcus bleeding index between the rats in the low - dose administration group and the model group, indicating that the low - dose drug had no effect on rat gingivitis. The gingival index and sulcus bleeding index of the rats in the high - dose administration group were significantly lower than those of the model group, indicating that the high - dose drug had a therapeutic effect on rat gingivitis. However, the low - dose group had no effect, indicating that the effect was not optimal.

[0068] Table 8 Gingival Index and Sulcus Bleeding Index of Rats with Gingivitis

[0069] ** P <0.01, **** P <0.0001, compared with the model group Example 3 was tested. In the gingivitis rat model, whether in the low - dose administration group or the high - dose administration group, the gingival index and sulcus bleeding index of the rats were significantly lower than those of the model group, and the gingival index and sulcus bleeding index of the rats in the high - dose group and the low - dose group were similar and there was no significant difference, indicating that the therapeutic effect of the low - dose drug on rat gingivitis had reached the upper limit. The results indicate that the drug has a therapeutic effect on gingivitis, but the effect is not as good as the optimal plan.

[0070] Table 9 Gingival Index and Sulcus Bleeding Index of Rats with Gingivitis

[0071] **** P <0.001, compared with the model group Example 4 was tested. In the gingivitis rat model, whether in the low - dose administration group or the high - dose administration group, the gingival index and sulcus bleeding index of the rats were significantly lower than those of the model group, and there was a dose - dependence. The results indicate that the drug has a therapeutic effect on gingivitis, and the drug combination is superior to Examples 2 and 3.

[0072] Table 10 Gingival Index and Sulcus Bleeding Index of Rats with Gingivitis

[0073] **** P <0.001, compared with the model group The present invention provides a drug that has the effects of preventing and improving gingivitis and periodontitis, reducing the gingival index, sulcus bleeding index and plaque index, and reducing the probing depth of periodontitis, and can effectively relieve and treat gingivitis and periodontitis; and has the effects of relieving gingival bleeding and inhibiting dental plaque.

[0074] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, those of ordinary skill in the art should understand that they can still modify the technical solutions described in the foregoing embodiments, or perform equivalent replacements on some of the technical features; and these modifications or replacements do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the various embodiments of the present invention.

Claims

1. A pharmaceutical composition, characterized in that The pharmaceutical composition comprises the following raw materials in parts by weight: 2-6 parts of white peony root, 0.5-4 parts of ginseng, 0.5-4 parts of mint, and 0.2-3 parts of liquorice.

2. The pharmaceutical composition according to claim 1, comprising the following raw materials in parts by mass: 2 to 5 parts of white peony root, 0.8 to 3 parts of ginseng, 0.8 to 3 parts of mint, and 0.3 to 2 parts of licorice.

3. The pharmaceutical composition according to claim 2, characterized in that The pharmaceutical composition comprises the following raw materials in parts by weight: 2.5-3.5 parts of white peony root, 1-1.5 parts of ginseng, 1-1.5 parts of mint, and 0.5-1 part of liquorice.

4. The pharmaceutical composition according to claim 3, characterized in that The raw materials of the pharmaceutical composition are 3 parts of white peony root, 1-1.2 parts of ginseng, 1-1.2 parts of mint, 0.5-0.6 parts of liquorice, and 55-100 parts of water.

5. The method for preparing the pharmaceutical composition according to any one of claims 1 to 4, characterized in that: include: 1) Extraction: Mix white peony root, ginseng, mint, licorice and water, decoct and filter to obtain an extract; 2) Concentration: Concentrating the extract under reduced pressure to obtain a concentrated solution; 3) Drying: spray drying the concentrate.

6. The method for preparing the pharmaceutical composition according to claim 5, characterized in that: In step 1), the decoction is performed at least twice, 6 to 10 times the amount of water is added for the first decoction, the decoction time is 1 to 2 hours, and the mixture is filtered; 4 to 8 times the amount of water is added for the second decoction, and the decoction time is 0.5 to 1.5 hours.

7. The method for preparing the pharmaceutical composition according to claim 6, characterized in that: In step 2), the temperature of the reduced pressure concentration is 55-75° C., the pressure is -0.08--0.04 MPa; and the relative density of the concentrated solution is 1.08-1.

15.

8. The method for preparing the pharmaceutical composition according to claim 6 or 7, characterized in that: In step 3), the air inlet temperature is 160~190°C, the air outlet temperature is 70~100°C, the nebulizer speed is 40~50HZ, and the infusion pump flow rate is 7~15HZ.

9. A medicament, characterized in that: A pharmaceutical composition comprising the pharmaceutical composition of any one of claims 1 to 4 or a pharmaceutical composition obtained by the method for preparing the pharmaceutical composition of any one of claims 5 to 8; preferably, the dosage form of the pharmaceutical composition is powder, paste, tablet, pill, capsule, granule, granule or oral solution.

10. Use of the pharmaceutical composition according to any one of claims 1 to 5, the pharmaceutical composition obtained by the preparation method according to any one of claims 5 to 8, or the medicament according to claim 9 in the preparation of a medicament for preventing and treating gingivitis and / or periodontitis.

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