CN113876891A

CN113876891A - Pharmaceutical composition for oral care and preparation method thereof

Info

Publication number: CN113876891A
Application number: CN202111291449.4A
Authority: CN
Inventors: 董金呈
Original assignee: Shandong Kangke Medical Instrument Co ltd
Current assignee: Shandong Kangke Medical Instrument Co ltd
Priority date: 2021-11-03
Filing date: 2021-11-03
Publication date: 2022-01-04

Abstract

The invention belongs to the technical field of medicines, and particularly relates to a pharmaceutical composition for oral care and a preparation method thereof. The pharmaceutical composition mainly takes anoectochilus roxburghii extract, hypochlorous acid and tea polyphenol as active ingredients, has excellent oral cavity cleaning effect, can improve oral pain and accelerate ulcer healing and regression, has obvious curative effect, is safe and reliable, has no toxic or side effect, and has wide clinical application prospect.

Description

Pharmaceutical composition for oral care and preparation method thereof

Technical Field

The invention belongs to the technical field of medicines, and particularly relates to a pharmaceutical composition for oral care and a preparation method thereof.

Background

Oral ulcer, commonly known as "aphtha", is a common ulcerative injury disease occurring in oral mucosa, which is mostly seen on mucosa of the inner side of the lip, tongue abdomen, buccal mucosa, vestibular sulcus, soft palate and other parts, and is a round or oval pain ulcer point. The oral ulcer has severe pain and obvious local burning pain during the attack, serious patients can influence diet and speaking, the daily life is greatly inconvenient, and the oral ulcer can be complicated with the general symptoms of halitosis, chronic pharyngitis, constipation, headache, dizziness, nausea, hypodynamia, dysphoria, fever, lymphadenectasis and the like. At present, the main treatment method of oral ulcer in China is local medication, and traditional Chinese medicine component medicines or glucocorticoid medicines with the functions of resisting inflammation, easing pain and promoting ulcer healing are locally used.

The hand-foot-and-mouth disease is a common infectious disease caused by various enteroviruses (EV71, CVA16 and the like), is mainly characterized by fever and rash or herpes at the parts of hands, feet, oral cavity and the like, and is mostly generated in children under the age of 5. Most patients will have fever when they get ill, and the symptoms of headache, cough, running nose and the like are accompanied, the course of disease is long, and the patients are very painful. According to the report of the official epidemic situation of infectious diseases of the CDC of China, more than one million cases of hand-foot-and-mouth diseases occur every year, and the diseases are the first of class C infectious diseases. At present, no specific medicine is available for clinical treatment of patients with hand-foot-and-mouth disease, and a symptomatic auxiliary treatment scheme is generally adopted according to clinical stages. The children patients are usually anorexia due to pain caused by oral mucosa rash or ulcer, how to quickly heal the oral herpes has important significance for improving the diet and nutrition of the children patients, and the discovery and development of the medicine which can be used for treating the hand-foot-and-mouth disease infection and has less side effect and high curative effect are urgently needed in the medical field.

Anoectochilus formosanus extract is well-known in terms of gold grass, magical drug, bird ginseng and the like, and has a wide treatment range in folk. The records of Fujian medicine record: the medicine has mild and sweet nature and flavor, has the effects of clearing heat and cooling blood, and dispelling wind and promoting diuresis, is mainly used for treating hemoptysis, bronchitis, nephritis, cystitis, diabetes, chyluria, hematuria, rheumatic arthritis, acute infantile convulsion, venomous snake bite and the like, and is mainly used for treating infantile hyperpyrexia and infantile convulsion in folk. The Chinese herbal medicine assembly means that the medicine has sweet taste and mild nature, has the effects of clearing heat and cooling blood, and removing dampness and detoxifying, and can treat pulmonary tuberculosis hemoptysis, diabetes, nephritis, cystitis, myasthenia gravis, spermatorrhea, rheumatic and rheumatoid arthritis, infantile convulsion, female leucorrhea, venomous snake bite and other symptoms. Modern pharmacology shows that the anoectochilus formosanus contains alkaloid, amino acid, saccharide, saponin, steroid and other components, and modern pharmacology research proves that the anoectochilus formosanus has the pharmacological activity effects of reducing blood sugar, reducing blood fat, protecting liver, resisting inflammation, easing pain, promoting urination, calming, reducing blood pressure, resisting oxidation, improving osteoporosis and the like.

At present, fresh anoectochilus formosanus, dried anoectochilus formosanus and anoectochilus formosanus tea are mainly sold in the market; in addition, a small amount of anoectochilus formosanus compound oral liquid and spray are used as hospital preparations for treating diseases of patients, but the hospital preparations are only used for treating the diseases of the patients in the hospital and are not suitable for daily medical care. Through retrieval, no anoectochilus formosanus preparation with a certain simple formula has an excellent oral bacteriostatic effect.

Disclosure of Invention

In order to make up for the defects of the prior art, the inventor provides a pharmaceutical composition for oral care through years of accumulated research and on the basis of inherited experiences and by combining with modern medical technology, the composition has an excellent oral cleaning effect, can improve oral pain and accelerate ulcer healing and regression, and has the advantages of remarkable curative effect, safety, reliability, no toxic or side effect and wide clinical application prospect.

In order to achieve the purpose, the technical scheme of the invention is as follows:

the pharmaceutical composition for oral care is characterized in that the active ingredients comprise anoectochilus formosanus extract, preferably, the anoectochilus formosanus extract accounts for 0.01-1.0% of the total volume of the pharmaceutical composition in percentage by volume; further preferably, the anoectochilus formosanus extract accounts for 0.05-0.5% of the total volume of the pharmaceutical composition in percentage by volume; further preferably, the anoectochilus formosanus extract accounts for 0.1-0.3% of the total volume of the pharmaceutical composition in percentage by volume.

The preparation method of the anoectochilus formosanus extracting solution comprises the following steps: adding 16-20 times of purified water into dry anoectochilus formosanus, soaking for 10-20min, heating and extracting for 20-40min in a water bath at 50-70 ℃, continuously extracting for two times, combining filtrates, and concentrating the anoectochilus formosanus extract to 0.5-0.7 g/mL.

In order to enhance the effect of the present invention, the pharmaceutical composition for oral care described above is characterized in that the active ingredients further comprise hypochlorous acid and tea polyphenols, the hypochlorous acid is present in the form of a solution, and various concentrations of hypochlorous acid can be used in the pharmaceutical composition of the present invention.

Tea Polyphenols (TP) are a complex of more than 30 phenolic compounds extracted from tea leaves. A large number of researches find that the tea polyphenol has the effects of resisting oxidation, resisting inflammation, resisting oxygen free radicals and the like.

Hypochlorous acid is an oxyacid of chlorine, is mainly used as a disinfectant, and is widely used for disinfecting polluted objects such as object surfaces, fabrics and the like, water, fruits, vegetables, eating and drinking utensils and the like. In addition to the above uses, hypochlorous acid can also be used for disinfection of indoor air, surfaces of secondary water supply facilities, hands, skin and mucous membranes.

The pharmaceutical composition can be added with pharmaceutically acceptable additives to prepare external preparations, such as spray, solution, liniment and the like, and preferably, the pharmaceutical composition is prepared into spray.

For the preparation and use convenience, the invention preferably uses the hypochlorous acid aqueous solution with the concentration of 50-300 ppm.

In some embodiments of the present invention, 80ppm hypochlorous acid aqueous solution is used, preferably, 20 mg-500 mg tea polyphenol and 100-500ml hypochlorous acid aqueous solution with 80ppm concentration are added to 1ml anoectochilus formosanus extract; more preferably, 50-100 mg of tea polyphenol and 400ml of hypochlorous acid aqueous solution with the concentration of 80ppm are added to 1ml of anoectochilus formosanus extract.

The invention also provides a medicinal composition spray for oral care, and the preparation method comprises the following steps:

(1) adding 16-20 times of purified water into dry anoectochilus formosanus, soaking for 10-20min, heating and extracting for 20-40min in a water bath at 50-70 ℃, continuously extracting for two times, combining filtrates, and concentrating the anoectochilus formosanus extract to 0.5-0.7 g/mL.

(2) Dissolving tea polyphenols in 50-100 mL of purified water, mixing with herba Anoectochili Roxburghii extract and hypochlorous acid water, adding purified water to desired concentration, and bottling.

In order to further explain the advantageous effects of the present invention, the inventors conducted a number of experimental studies. The method comprises the following steps: the preparation method of the composition is provided in the part of the pharmaceutical preparation (examples 1-9), and the performance test results of the comparative examples 1-2 show that the spray prepared by the examples of the invention is very stable; animal test example 1 shows that the pharmaceutical composition of the present invention is safe and non-irritating to skin contact; animal experiment example 2 shows that the pharmaceutical composition of the invention can promote the healing of oral ulcer of rats; animal experiment example 3 shows that the pharmaceutical composition of the present invention can inhibit hand-foot-and-mouth disease virus in vitro, and therefore, the present invention also claims the use of the above composition in the preparation of drugs for preventing or treating dental ulcer or hand-foot-and-mouth disease.

Compared with the prior art, the invention has the beneficial effects that: the oral liquid has a simple formula, has an excellent oral cleaning effect, can improve oral pain and accelerate ulcer healing and resolution, and has the advantages of obvious curative effect, safety, reliability and no toxic or side effect.

Detailed Description

The present invention will be further described below by way of specific embodiments, but the present invention is not limited to only the following examples. It will be apparent to those skilled in the art that the invention can be modified and replaced with other components having the same effects without departing from the spirit and scope of the invention, and all such modifications and substitutions are deemed to be within the scope of the invention.

Preparation example

Example 1

Prescription: 300mL of hypochlorous acid water with the concentration of 80ppm, 1mL of anoectochilus formosanus extract, 60mg of tea polyphenol and purified water which are added to 750 mL

The preparation method comprises the following steps: adding 18 times of purified water into dried herba Anoectochili Roxburghii, soaking for 15 min, heating in water bath at 60 deg.C for 30min, continuously extracting twice, mixing filtrates, and concentrating herba Anoectochili Roxburghii extract to 0.5 g/mL.

Dissolving 60mg of tea polyphenol in 100 mL of purified water, mixing uniformly with 1mL of anoectochilus formosanus extract and 300mL of hypochlorous acid water with the concentration of 80ppm, adding the purified water to 750 mL, and filling into a spray bottle to obtain the tea polyphenol.

Example 2

Prescription: 250mL of hypochlorous acid water with the concentration of 80ppm, 1.1mL of anoectochilus formosanus extract, 70mg of tea polyphenol and purified water which are added to 750 mL

The preparation method comprises the following steps: adding 18 times of purified water into dried Anoectochilus roxburghii, soaking for 15 min, heating in 65 deg.C water bath for 30min, continuously extracting twice, mixing filtrates, and concentrating Anoectochilus roxburghii extract to 0.5 g/mL for use.

Dissolving tea polyphenols 70mg in 100 mL of purified water, mixing well with herba Anoectochili Roxburghii extract 1.1mL and hypochlorous acid water 250mL with concentration of 80ppm, adding purified water to 750 mL, and filling into spray bottle.

Example 3

Prescription: 350mL of hypochlorous acid water with the concentration of 80ppm, 1mL of anoectochilus formosanus extract, 80mg of tea polyphenol and purified water which are added to 750 mL

The preparation method comprises the following steps: adding 17 times of purified water into dried herba Anoectochili Roxburghii, soaking for 20min, heating in 65 deg.C water bath for 30min, continuously extracting for two times, mixing filtrates, and concentrating herba Anoectochili Roxburghii extract to 0.6 g/mL.

Dissolving tea polyphenols 80mg in 100 mL of purified water, mixing well with herba Anoectochili Roxburghii extract 1mL and hypochlorous acid water 350mL with concentration of 80ppm, adding purified water to 750 mL, and bottling.

Example 4

Prescription: 350mL of hypochlorous acid water with the concentration of 80ppm, 1.5mL of anoectochilus formosanus extract, 60mg of tea polyphenol and purified water which are added to 750 mL

The preparation method comprises the following steps: adding 16 times of purified water into dried Anoectochilus roxburghii, soaking for 15 min, heating in water bath at 60 deg.C for 35 min, continuously extracting twice, mixing filtrates, and concentrating Anoectochilus roxburghii extract to 0.55 g/mL for use.

Dissolving 60mg of tea polyphenol in 50mL of purified water, mixing uniformly with 1.5mL of anoectochilus formosanus extract and 350mL of hypochlorous acid water with the concentration of 80ppm, adding purified water to 750 mL, and filling into a spray bottle to obtain the tea polyphenol.

Example 5

Prescription: 400mL of hypochlorous acid water with the concentration of 80ppm, 1.5mL of anoectochilus formosanus extract, 90mg of tea polyphenol and purified water which are added to 1000 mL

The preparation method comprises the following steps: adding 18 times of purified water into dried herba Anoectochili Roxburghii, soaking for 20min, heating in water bath at 60 deg.C for 30min, continuously extracting twice, mixing filtrates, and concentrating herba Anoectochili Roxburghii extract to 0.5 g/mL.

Dissolving 90mg of tea polyphenol in 120 mL of purified water, mixing uniformly with 1.5mL of anoectochilus formosanus extract and 400mL of hypochlorous acid water with the concentration of 80ppm, adding purified water to 1000 mL, and filling into a spray bottle to obtain the tea polyphenol.

Example 6

Prescription: 700mL hypochlorous acid water with the concentration of 80ppm, 2mL anoectochilus formosanus extract, 130mg tea polyphenol and purified water which are added to 1200 mL

The preparation method comprises the following steps: adding 18 times of purified water into dried Anoectochilus roxburghii, soaking for 15 min, heating in water bath at 60 deg.C for 30min, continuously extracting twice, mixing filtrates, and concentrating Anoectochilus roxburghii extract to 0.65 g/mL for use.

Dissolving 130mg of tea polyphenol in 200mL of purified water, mixing uniformly with 2mL of anoectochilus formosanus extract and 700mL of hypochlorous acid water with the concentration of 80ppm, adding the purified water to 1200 mL, and filling into a spray bottle to obtain the tea polyphenol.

Example 7

Prescription: 500mL of hypochlorous acid water with the concentration of 80ppm, 1.5mL of anoectochilus formosanus extract, 100mg of tea polyphenol and purified water which are added to 1000 mL

The preparation method comprises the following steps: adding 16 times of purified water into dried herba Anoectochili Roxburghii, soaking for 15 min, heating in water bath at 60 deg.C for 25min, continuously extracting twice, mixing filtrates, and concentrating the herba Anoectochili Roxburghii extract to 0.5 g/mL.

Dissolving tea polyphenols 100mg in 100 mL of purified water, mixing with herba Anoectochili Roxburghii extract 1.5mL and hypochlorous acid water 500mL with concentration of 80ppm, adding purified water to 1000 mL, and bottling.

Example 8

Prescription: 400mL of hypochlorous acid water with the concentration of 100ppm, 1.5mL of anoectochilus formosanus extract, 130mg of tea polyphenol and purified water are added to 1000 mL

Dissolving 130mg of tea polyphenol in 100 mL of purified water, mixing uniformly with 1.5mL of anoectochilus formosanus extract and 400mL of hypochlorous acid water with the concentration of 100ppm, adding purified water to 1000 mL, and filling into a spray bottle to obtain the tea polyphenol.

Example 9

Prescription: 200mL of hypochlorous acid water with the concentration of 200ppm, 2mL of anoectochilus formosanus extract, 150mg of tea polyphenol and purified water which are added to 1000 mL

The preparation method comprises the following steps: adding 18 times of purified water into dried herba Anoectochili Roxburghii, soaking for 20min, heating in water bath at 60 deg.C for 30min, continuously extracting twice, mixing filtrates, and concentrating herba Anoectochili Roxburghii extract to 0.6 g/mL.

Dissolving 150mg of tea polyphenol in 100 mL of purified water, mixing uniformly with 2mL of anoectochilus formosanus extract and 200mL of hypochlorous acid water with the concentration of 200ppm, adding purified water to 1000 mL, and filling into a spray bottle to obtain the tea polyphenol.

Comparative example 1

Prescription: 300mL of hypochlorous acid water with the concentration of 80ppm, 60mg of tea polyphenol and purified water which are added to 750 mL

The preparation method comprises the following steps: dissolving 60mg of tea polyphenol in 100 mL of purified water, mixing uniformly with 300mL of hypochlorous acid water with the concentration of 80ppm, and adding the purified water to 750 mL to obtain the tea polyphenol.

Comparative example 2

Prescription: 150mL of hypochlorous acid water with the concentration of 200ppm, 60mg of tea polyphenol and purified water added to 750 mL

The preparation method comprises the following steps: dissolving 60mg of tea polyphenol in 100 mL of purified water, mixing uniformly with 150mL of hypochlorous acid water with the concentration of 200ppm, and adding the purified water to 750 mL to obtain the tea polyphenol.

(II) performance test:

and (3) detecting according to a stability detection method in GB/T2738-2012 daily chemical product antibacterial and bacteriostatic effect evaluation. The test sample was placed in a brown glass bottle, hermetically stored in an incubator at 37 ℃ for 90 days, sampled, and the available chlorine content in the secondary sample bottle was measured by iodometry, and the stability thereof was evaluated by the decrease rate of the available chlorine content. The stability of examples 1-6 and comparative examples 1-2 is as follows:

the tests show that the spray prepared by the embodiment of the invention is very stable, according to the requirements of 'Disinfection technical Specification 2002 edition' on stability acceleration experiments, the preparation of the invention is stored for 90 days at constant temperature and constant humidity at 37 ℃, the reduction of available chlorine is less than or equal to 10%, the validity period can reach 2 years, and the stability of the sample of the comparative embodiment which does not contain anoectochilus formosanus extract is poor.

(III) animal test example

Example 1 evaluation of safety of the pharmaceutical composition of the present invention

(1) Acute toxicity test of skin

Selecting 12 healthy New Zealand white rabbits (weight 2.5-3.0 kg), male and female half, and hair-removing region with 8% sodium sulfide in bilateral symmetrical region of back spine, wherein the hair-removing region has a body surface area of 10% (about 150 cm)²) The breeding is carried out in cages, the observation is carried out for 24 hours, the breeding is randomly divided into 3 groups, and each group comprises 4 animals. Blank control group: 1g of physiological saline is coated on the spinal cord depilated area; ② complete skin group: 1g of the spray solution prepared in the embodiment 1 of the invention is only coated on the depilatory area on the two sides of the spinal column; (iii) damaged skin group: 1g of the spray solution prepared in the embodiment 1 of the invention is only coated on depilated areas on two sides of a spinal column, and the damaged skin is prepared by scratching a # shape on the depilated and disinfected skin by using a disinfection needle to the extent of blood seepage. And observing and recording the weight, skin, hair, eye and mucosa changes and respiratory, central nervous system and limb movement of each rabbit from 1, 24, 48, 72 h to 7d every day.

And (3) test results: all animals in each group survived, and no obvious abnormality was found in the animals in the drug-coated group in terms of ingestion, body weight, skin in the depilated area, conjunctiva of eyes and oral mucosa, respiration, circulation, central nervous system and limb activities. Therefore, the pharmaceutical composition has low toxicity to the skin of the New Zealand white rabbits, and the clinical medication safety of the pharmaceutical composition is prompted.

(2) Skin irritation test

20 New Zealand white rabbits were dehaired by 8% sodium sulfide on both sides of the spine 24h before administration, and the dehaired region was 10% of body surface area (about 150 cm)²) After unhairing, 24h, the skin was examined for injury due to unhairing. Dividing into 4 groups, namely an intact skin control group, an intact skin administration group, a damaged skin control group and a damaged skin administration group, marking # on the unhaired and sterilized skin by using a sterilized needle, and basically keeping the damaged degree of the skin on the left side and the right side consistent by taking the skin bleeding as a degree. Intact skin control group: 1g of physiological saline is added into the mixtureApplying to a spinal epilation area; ② complete skin administration group: 1g of the spray solution prepared in the embodiment 1 of the invention is only coated on the spinal epilation area; damaged skin control group: applying physiological saline 1 g/spinal cord depilatory area; (iv) damaged skin administration group: 1g of the spray solution prepared in example 1 of the present invention was applied to only the spinal depilation area. Observing after 24 hours; the same test substance was then applied for 3 consecutive days and the skin reaction in the depilated area was observed l, 24, 48, 72 h after the last application. The appearance of erythema and edema scores were recorded and the stimulation intensity was assessed.

And (3) test results: in the experiment, 1 skin of the white rabbit respectively shows slight erythema in the single-dose and multiple-dose damaged skin groups, but the skin can automatically regress within 48 hours, and no erythema or edema reaction is seen in other groups. The results were evaluated in terms of skin irritation intensity and showed that the formulation was not irritating to skin contact by New Zealand white rabbits.

Example 2: experimental study on promotion of spray to healing of oral ulcer of rat

1. Experimental animals: 25 clean Wistar rats with half male and half female, and the body mass of 180-220 g. 20 were randomly drawn for modeling and 5 were used for blank control.

And molding: a2% sodium pentobarbital anesthetized rat is subjected to a 5mm caliber treatment, a pasteur tube with cotton plugged in the tube orifice is used for sucking a saturated sodium hydroxide solution, and the pasteur tube orifice is attached to the mucous membrane of the lower lip of the rat for 60 s. And (3) after modeling for 24h, visually observing the oral mucosa of the rat to form an ulcer with the diameter of about 5mm, wherein the center of the ulcer is sunken, the surface of the ulcer is covered by a yellowish-white false membrane, and bleeding and hyperemia and edema are generated around the ulcer, so that the success of modeling is realized.

The grouping administration method comprises the following steps: the 20 rats were successfully modeled, and were randomly divided into a positive control group (coated with the rehabilitation new solution), a test group (coated with the spray solution prepared in example 1 of the present invention), a comparative test group (coated with the spray solution prepared in comparative example 1 of the present invention), and a model control group (coated with physiological saline) each of which was 10, and the respective drugs were dipped with sterilized cotton swabs and coated on the oral ulcer sites of SD rats for 3 min. The treatment was given 2 times daily for 7 consecutive days. At 1 d, 3 d, 5d and 7d after administration of the therapeutic drug, ulcer sites were traced with cellophane, ulcer area calculation was performed using image-pro plus 6.0 (medical image analysis software), and healing time was recorded.

And evaluating the degree of the oral ulcer: 0, indicating that the oral mucosa is in a normal state and has no ulcer; i, indicating that ulcer exists, but no pseudomembrane is obvious on the surface of the ulcer; II, showing that a thin yellow-white false membrane is arranged on the surface of the oral ulcer, but the periphery of the oral ulcer has no edema; III, showing the appearance of a thick pseudomembrane on the surface of the canker sore and the surrounding with a marked inflammatory edema manifestation; IV, thick pseudomembrane on the surface of canker sores with significant inflammatory edema around the sores.

And a statistical method comprises the following steps: performing multiple group comparison by using SPSS21.0 statistical software and analysis of variance, and using the result

S indicates that P < 0.05 indicates a statistical difference.

And the result is as follows:

(1) change in the area of traumatic ulcers in rats: compared to a placebo control group (no ulcer formation). The area comparison of the rats with the 1 d positive control group, the test group, the comparative test group and the model control group has no statistical difference (P is more than 0.05); 3-7 d, compared with the model control group, the areas of the oral ulcers of the rats seen in the positive control group, the comparative test group and the test group are reduced, and the difference has statistical significance (P is less than 0.05); compared with the positive control group, the test group and the positive control group have the advantages that the oral ulcer area of the rat is reduced, the difference has statistical significance (P is less than 0.05), and compared with the positive control group, the difference has no statistical significance (P is more than 0.05).

(2) Change in the degree of traumatic ulcer in rats: compared with a blank control group (no ulcer is formed in the blank control group), the rats of the 1 d positive control group, the test group, the contrast test group and the model control group have no statistical difference (P is more than 0.05) in the area comparison of the traumatic ulcer; 3 d, the degree of alleviating oral ulcer of the positive control group, the comparative test group and the experimental group is not statistically different (P is more than 0.05); in 5-7 d, the degree of oral ulcer in the experimental group is obviously reduced (P is less than 0.05).

(3) Time to heal traumatic ulcer in rats: compared among treatment groups, the ulcer in the 5d experimental group was substantially healed, the ulcer in the 7d experimental group and the comparative experimental group was substantially healed, and the ulcer in most of the rats in the model control group was healed at the 9 d. See in particular Table 1

Table 1: the influence of the spray of the invention on the degree of the canker sore of the rat is as follows:

example 3: test for in vitro inhibition of viruses by spray agent of the present invention

1. Experimental Material

Virus seeds: enterovirus 71 (EV71) and coxsackievirus A16 (CVA 16) are isolated from stool specimens of patients with hand-foot-and-mouth disease and provided by Fujian university of traditional Chinese medicine.

Cell: RD cells (provided by fujian university of medicine).

Experimental drugs: example 1.

Positive control: vidarabine monophosphate (produced by Hainan chemical combination pharmaceutical industry Co., Ltd.).

GIBCO RPMI1640 cell culture medium (containing 10% newborn bovine serum, purchased from ThermoFisher Co.).

GIBCO RPMI1640 cell maintenance solution (containing 2% newborn bovine serum, purchased from ThermoFisher Co.).

Experimental method

And (3) virus propagation: respectively inoculating EV71 and CVA16 viruses on a single layer of RD cells, adding a maintenance solution, culturing for 72 hours at 37 ℃ in a 5% carbon dioxide incubator to generate more than 90% of lesions, freezing and thawing for 3 times, blowing, centrifuging, quantitatively subpackaging, and placing in a refrigerator at minus 80 ℃ for freezing and storing for later use to obtain a virus solution.

Toxicity inhibition experiment: 10ml of the test drug (obtained in example 1), 10ml of the virus solution (prepared as described above) and 10ml of the cell maintenance solution (RPMI 1640, original concentration) were mixed in a ratio of 1: 1, placing the mixture at 37 ℃ for 1 hour, diluting the mixture by cell maintenance solutions respectively at 10-1, 10-2, 10-3, 10-4, 10-5, 10-6, 10-7, 10-8, 10-9, 10-10 and 10-11, longitudinally repeating the steps for 3 holes respectively, sequentially and transversely adding the diluted solutions to a monolayer of cells in a 96-well plate, and setting a positive control (12 columns in total, vidarabine monophosphate). The cells were incubated at 37 ℃ in a 5% carbon dioxide incubator, and the lesions were observed daily and after 5 days, the observation was terminated. Calculating the cell morbidity and the cell survival rate, calculating the virus virulence by using a Reed-Muench method, and observing the virus killing effect of the medicament. The virus titer was compared to the virus control, a titer reduction was effective, and half the cytotoxic concentration was divided by half the effective concentration to give the Therapeutic Index (TI), the results are shown in table 2. The results show that the experimental drug has direct inactivation effect on both EV71 virus and CVA16 virus, and the therapeutic indexes are respectively 4.8 times and 5.3 times of vidarabine monophosphate.

TABLE 2 determination of therapeutic index

Claims

1. A pharmaceutical composition for oral care, characterized in that the active ingredient comprises anoectochilus formosanus extract.

2. The pharmaceutical composition of claim 1, wherein the anoectochilus formosanus extract accounts for 0.01-1.0% of the total volume of the pharmaceutical composition in volume percent.

3. The pharmaceutical composition of claims 1-2, wherein the active ingredients of the pharmaceutical composition further comprise hypochlorous acid and tea polyphenols.

4. The pharmaceutical composition of claim 3, wherein the hypochlorous acid in the pharmaceutical composition is an aqueous solution of hypochlorous acid having a concentration of 50 to 300 ppm.

5. The pharmaceutical composition as claimed in claim 3, wherein the pharmaceutical composition comprises 20 mg-500 mg of tea polyphenols per 1ml Anoectochilus roxburghii extract, and 100 ml of hypochlorous acid aqueous solution with a concentration of 80 ppm.

6. The pharmaceutical composition of claim 5, wherein the pharmaceutical composition is formulated as a topical dosage form.

7. The pharmaceutical composition of claim 5, wherein the pharmaceutical composition is formulated as a spray.

8. The pharmaceutical composition of claims 1-2, wherein the preparation method of the anoectochilus formosanus extract comprises the following steps: adding 16-20 times of purified water into dry anoectochilus formosanus, soaking for 10-20min, heating and extracting for 20-40min in a water bath at 50-70 ℃, continuously extracting for two times, combining filtrates, and concentrating the anoectochilus formosanus extract to 0.5-0.7 g/mL.

9. The pharmaceutical composition of claim 4, wherein the pharmaceutical composition spray is prepared by a method comprising the steps of:

(1) adding 16-20 times of purified water into dry Anoectochilus roxburghii, soaking for 10-20min, heating and extracting in a water bath at 50-70 ℃ (60 ℃) for 20-40min, continuously extracting for two times, combining filtrates, and concentrating Anoectochilus roxburghii extract to 0.5-0.7 g/mL;

(2) dissolving tea polyphenols in purified water, mixing with herba Anoectochili Roxburghii extract and hypochlorous acid water, adding purified water to desired amount, and packaging into spray bottle.