CN106913627B

CN106913627B - Infantile Pudilan anti-inflammation syrup and preparation method thereof

Info

Publication number: CN106913627B
Application number: CN201710164692.7A
Authority: CN
Inventors: 曹龙祥; 董自波; 田刚; 李超; 邵建国; 李萍
Original assignee: Jumpcan Pharmaceutical Group Co ltd
Current assignee: Jumpcan Pharmaceutical Group Co ltd
Priority date: 2017-03-20
Filing date: 2017-03-20
Publication date: 2021-01-05
Anticipated expiration: 2037-03-20
Also published as: CN106913627A

Abstract

The invention relates to a traditional Chinese medicine extract, a medicinal preparation thereof and a preparation method thereof, in particular to a pediatric Pudilan anti-inflammatory syrup and a preparation method thereof. The invention provides a preparation method of Pudilan extract with anti-inflammatory effect, which comprises the steps of extracting raw material medicines of dandelion 3.0-5.0 parts, isatis root 1.0-2.0 parts, corydalis bungeana 0.5-1.5 parts and scutellaria baicalensis 1.0-2.0 parts by weight with water, and then precipitating with alcohol. The method for preparing the Pudilan extract adopts water extraction of the whole formula, has simple process and is beneficial to production. The pediatric Pudilan anti-inflammatory syrup prepared by the invention has good stability, good taste and high bioavailability. The medicine has effects of clearing away heat and toxic materials, and relieving inflammation or swelling, and can be used for treating parotitis, pharyngitis and tonsillitis.

Description

Infantile Pudilan anti-inflammation syrup and preparation method thereof

Technical Field

The invention relates to a traditional Chinese medicine extract, a medicinal preparation thereof and a preparation method thereof, in particular to a pediatric Pudilan anti-inflammatory syrup and a preparation method thereof.

Background

Epidemic parotitis, acute pharyngitis and tonsillitis are common diseases of children, and the epidemic parotitis belongs to infectious diseases and is commonly seen in bacterial and viral diseases. The traditional Chinese medicine is named as mumps; the Huyonmei summarizes that the incidence rates of epidemic parotitis in the Chinese 2010-2012 epidemic parotitis disease characteristic analysis are 22.3963/10 ten thousand, 33.8363/10 ten thousand and 35.5897/10 ten thousand respectively within 3 consecutive years, wherein the ages below 15 years account for 91.58%, 90.93% and 90.27% of the total cases in the year. Tonsillitis belongs to upper respiratory tract infection, and has incomplete immune system and lower resistance than adults because the respiratory system of children is immature. At present, acute upper respiratory tract infection is the first infectious disease of children, the incidence rate of acute tonsillitis is the highest, virus infection is the most common, the traditional Chinese medicine belongs to the category of tonsillitis, the traditional Chinese medicine is caused by invasion of exogenous pathogenic factors, invasion of the throat or excessive heat in the lung and the stomach and attack of the throat, the pediatric symptoms are more serious than those of adults, and the pediatric symptoms are often accompanied by high fever. Pharyngitis also belongs to upper respiratory tract infection, and the incidence rate is higher in children, especially acute pharyngitis, which is acute inflammation of pharyngeal mucosa and submucosal tissues. The traditional Chinese medicine considers that the acute pharyngitis belongs to the category of acute pharyngitis, the cause is similar to the acute tonsillitis, the viscera are mainly diseased in the lung and the stomach, and the wind-heat syndrome and the lung-stomach heat excess syndrome are common clinically.

At present, western medicine mostly treats the diseases with antibiotics, and the treatment of the diseases by traditional Chinese medicine is superior along with the increasing severity of antibiotic resistance. Although antibiotic treatment is really effective, the side effects are large and relapse is easy. The traditional Chinese medicine considers that the heat clearing and detoxifying, the detumescence and the sore throat relieving are suitable for treating the diseases, the curative effect of the therapy is safe and reliable, the side effect is less, and the popularization value is high. The Pudilan anti-inflammatory oral liquid is a marketed variety, and has the effects of clearing away heat and toxic materials, relieving swelling and relieving sore throat. Can be used for treating furuncle, parotitis, pharyngitis, and tonsillitis. Years of clinical application show that the strain has a better market in pediatrics, but the usage and dosage of the Pudilan anti-inflammatory oral liquid do not specifically refer to the usage and dosage of children, and only the dosage of children is reduced. In the clinical use process, the oral liquid is found to have poor taste for children and difficult administration for children, so that the development of a new medicinal preparation suitable for children to take is particularly important on the basis. The syrup is concentrated sucrose aqueous solution containing medicine or aromatic substances for oral administration, and sucrose, aromatic, etc. can mask unpleasant odor of medicine, improve taste, and is especially popular with children. The syrup is easy to be polluted by microorganisms, the main additive in the syrup is preservative, and sensitive people in special period of physical development such as children and pregnant women are not suitable for eating the food which excessively uses the preservative. Meanwhile, in the process of the Pudilan anti-inflammatory oral liquid, the scutellaria baicalensis needs to be extracted independently, and the process is complex.

Disclosure of Invention

The invention aims to provide a Pudilan extract with an anti-inflammatory effect and a preparation method thereof.

The invention also aims to provide the pediatric Pudilan anti-inflammatory syrup and the preparation method thereof, and the syrup has good taste, good stability and high bioavailability.

The invention also aims to provide application of the pediatric Pudilan anti-inflammatory syrup medicament.

The invention provides a preparation method of a Pudilan extract with an anti-inflammatory effect, which comprises the following steps:

1) water extraction: 3.0-5.0 parts of dandelion, 1.0-2.0 parts of isatis root, 0.5-1.5 parts of bunge corydalis herb and 1.0-2.0 parts of scutellaria baicalensis are extracted by water, wherein after the dandelion, the isatis root and the bunge corydalis herb are boiled in water, the scutellaria baicalensis is put into the water and extracted by the water to obtain water extract;

2) alcohol precipitation: concentrating the water extractive solution to relative density of 60-70 deg.C, measuring with heat at 1.10-1.15, adding anhydrous ethanol to make ethanol volume content in the concentrated water extractive solution be 60%, filtering to remove insoluble substances, and removing solvent from the filtrate to obtain extract.

Preferably, the raw material medicines comprise 4 parts of dandelion, 1.5 parts of isatis root, 1.0 part of corydalis bungeana and 1.5 parts of scutellaria baicalensis

The invention relates to a water extraction of a whole formula, which is prepared by extracting four raw material medicines in a formula: the dandelion, the isatis root, the corydalis bungeana and the scutellaria baicalensis are extracted by water at one time, and the scutellaria baicalensis is not required to be extracted by water separately.

The weight of the water for water extraction is 10 times of that of the decocted raw material medicine, the water is decocted for 3 times, each time lasts for 1.5 hours, and the decocted liquid of each time is combined.

The time for placing the mixture after adding the ethanol in the alcohol precipitation step is preferably 24 hours.

The invention provides a Pudilan extract with anti-inflammatory effect, which is prepared by the method.

The invention provides another technical scheme for providing a Pudilan anti-inflammatory drug, which consists of the extract prepared by the method as an active ingredient and pharmaceutically acceptable auxiliary materials.

The Pudilan anti-inflammatory drug is syrup, namely Pudilan anti-inflammatory syrup.

The sweetening agent contained in the Pudilan anti-inflammatory syrup is one or more of sucrose, stevioside and sucralose;

preferably, the sweeteners contained in the pudendum blue anti-inflammatory syrup are sucrose, steviol glycosides and sucralose;

more preferably, the sweetener content in the Pudilan anti-inflammatory syrup is 45% (g/mL) sucrose, 0.2% (g/mL) steviol glycoside and 0.1% (g/mL) sucralose.

The Pudilan anti-inflammatory drug syrup also contains essence;

preferably, the essence is orange essence;

more preferably, the cattleya blue syrup comprises 0.15% (g/mL) orange flavour.

The Pudilan anti-inflammatory syrup is prepared from 500g of dandelion, 188g of isatis root, 125g of corydalis bungeana and 188g of scutellaria baicalensis, and the preparation method comprises the following steps:

1) water extraction: boiling herba Taraxaci, radix Isatidis and herba corydalis Bungeanae in water, adding Scutellariae radix, and extracting with water to obtain water extractive solution;

2) alcohol precipitation: concentrating the extracting solution to obtain a clear paste with the relative density of 60-70 ℃ and the thermal test temperature of 1.10-1.15, adding absolute ethyl alcohol to ensure that the volume content of the ethyl alcohol in the concentrated water extracting solution is 60%, standing for 24h, filtering, and removing the ethyl alcohol from the supernatant to obtain an extract with the relative density of 60-70 ℃ and the thermal test temperature of 1.15-1.20;

3) adding water to the extract to 700mL, refrigerating for 48h, filtering, adjusting the pH value of the supernatant to 6.5 with sodium hydroxide solution, adding 450g of sucrose, 2g of stevioside and 1g of sucralose, boiling, filtering, adding 1.5g of orange essence, and adding water to 1000 mL.

The Pudilan anti-inflammatory syrup prepared by the invention is not added with a stabilizing agent and a preservative, and is clear, good in stability and high in bioavailability.

The Pudilan anti-inflammatory syrup prepared by the invention has good taste, improves the compliance of children and is suitable for children to take.

The stability of the cichoric acid is influenced by various factors such as pH value, temperature and the like, and the baicalin is dissolved in the alkali solution, but is unstable and changes dark brown gradually in the alkali solution.

The extract prepared by the invention is applied to preparing the medicine for treating infantile epidemic parotitis, acute pharyngitis or acute tonsillitis.

The preparation method of the extract provided by the invention adopts water extraction of the whole formula, the process is simple, the production is facilitated, and the pediatric Pudilan anti-inflammatory syrup prepared from the extract has the advantages of good stability, clarity, transparency, good taste, high bioavailability, stable preparation process, greenness and energy conservation.

Drawings

Fig. 1 is a photograph showing pathological histological observation of pharynx.

Detailed Description

The present invention will be further understood from the specific examples of the present invention and comparative examples given below. They are not intended to limit the invention.

Example 1

1 Water extraction of medicinal materials

1.1 orthogonal preferential aqueous extraction Process

Performing L on the water addition amount, the extraction time and the extraction times according to three factors and three levels₉(3⁴) The orthogonal design preferably selects the optimal water extraction process condition.

TABLE 1 Water extraction orthogonal test factor horizon

Taking 50g of dandelion, 18.8g of isatis root, 12.5g of corydalis bungeana and yellow18.8g of radix Scutellariae, 9 parts each by L₉(3⁴) And performing an orthogonal table test, and preferably selecting an optimal water extraction process by taking the contents of baicalin and chicoric acid as indexes, wherein the results are shown in tables 2-4.

TABLE 2 orthogonal test design and results table

TABLE 3 analysis of variance of baicalin extracted from water

TABLE 4 analysis of variance of chicoric acid extracted from water

The influence orders of the factors of the result are visually analyzed by taking the contents of the baicalin and the chicoric acid as indexes, and the influence orders are C>B>A; the baicalin variance analysis result shows that the factor C has extremely significant influence, and the optimal extraction process is A₂B₃C₃Namely, 10 times of water is added, and extraction is carried out for three times, 2h each time. The chicoric acid variance analysis result shows that the factor C has significant difference, and the optimal extraction process is A₃B₃C₃Namely, 12 times of water is added, and extraction is carried out for three times, 2h each time. Due to B₃And B₂The difference of the baicalin content is small; a. the₃And A₂，B₃And B₂The content of chicoric acid is slightly different. Comprehensively considering, the optimal process is determined as follows: a. the₂B₂C₃Namely, 10 times of water is added for 3 times of extraction, and each time lasts for 1.5 h.

1.2 Water extraction verification Process

250g of dandelion, 94g of radix isatidis, 62.5g of corydalis bungeana and 94g of scutellaria baicalensis are respectively taken and subjected to 3 verification tests according to the optimal scheme optimized by orthogonal tests.

TABLE 5 Water extraction verification test results

From the above results, 3 times of verification showed that the average baicalin content was 87.0 mg/g-crude drug and RSD was 0.78%, and the average chicoric acid content was 0.899 mg/g-crude drug and RSD was 2.47%, indicating that the preferred water extraction process was stable and feasible.

Example 2

1 Water extraction and alcohol precipitation optimization

1.1 orthogonal optimization alcohol precipitation process

Performing L treatment on the relative density, alcohol precipitation concentration and alcohol precipitation time of the extract according to three factors and three levels₉(3⁴) The optimal alcohol precipitation process is optimized through orthogonal design.

TABLE 6 level table of factors of alcohol precipitation orthogonal test

Filtering the extracting solution verified by water extraction, merging, concentrating until the relative density is 1.05-1.10, and averagely dividing into 3 parts: the first part is divided into 3 parts on average; the second part is continuously concentrated until the relative density is 1.10-1.15, and then is averagely divided into 3 parts; and the third part is continuously concentrated until the relative density is 1.15-1.20, and then is averagely divided into 3 parts. According to L₉(3⁴) And performing an alcohol precipitation test on the orthogonal test table, and performing visual analysis and variance analysis by taking the contents of the baicalin and the chicoric acid as investigation indexes, wherein the results are shown in tables 7-9.

Table 7 alcohol precipitation orthogonal test table

TABLE 8 analysis of variance of alcohol precipitated baicalin

TABLE 9 analysis of variance of alcohol precipitated chicoric acid

The influence orders of the factors of the result are visually analyzed by taking the contents of the baicalin and the chicoric acid as indexes, wherein the influence orders are B>A>C; the baicalin variance analysis result shows that the factor B has significant influence, the factor A, C has no influence, and the optimal alcohol precipitation process is A₁B₁C₂The relative density of the extract is 1.05-1.10, the alcohol precipitation concentration is 60%, and the alcohol precipitation is carried out for 24 h. The variance analysis result of the chicoric acid shows that the factor B has very significant difference, the factor A, C has no influence, and the optimal alcohol precipitation process is A₂B₁C₃The relative density of the extract is 1.10-1.15, the alcohol precipitation concentration is 60%, and the alcohol precipitation is carried out for 48 h. Due to A₁The influence on the alcohol precipitation process is large, the precipitate is not firm and is difficult to filter, the comprehensive consideration is carried out, and finally the alcohol precipitation process is determined to be A₂B₁C₂Namely, the relative density of the extract is 1.10-1.15, the alcohol precipitation concentration is 60%, and the alcohol precipitation is carried out for 24 h.

1.2 alcohol precipitation verification process

Taking 300g of dandelion, 112.8g of isatis root, 75g of corydalis bungeana and 112.8g of scutellaria baicalensis, extracting 3 parts of dandelion, 112.8g of isatis root, 75g of corydalis bungeana and 112.8g of scutellaria baicalensis according to an optimal water extraction process, and performing 3 times of alcohol precipitation verification tests according to an optimal scheme optimized by an alcohol precipitation orthogonal test.

TABLE 10 alcohol precipitation verification test results

From the results, the 3-time alcohol precipitation verification proves that the average content of baicalin is 52.74mg/g, the RSD is 1.24%, the average content of chicoric acid is 0.652mg/g, and the RSD is 0.94%, which indicates that the preferable process of alcohol precipitation is stable and feasible.

Example 3

1. Screening for refrigeration time

On the basis of a pre-experiment, taking a certain amount of alcohol precipitation supernatant, concentrating until the relative density is 1.15-1.20, averagely dividing into four parts, respectively adding water to 70% of the amount of the prescription, respectively refrigerating and standing for 24h, 36h, 48h and 72h, comparing the contents of baicalin and chicoric acid, and simultaneously inspecting the clarity of a sample.

TABLE 11 screening results for refrigeration time

From the above results, it can be seen that the contents of baicalin and chicoric acid are gradually reduced with the increase of the refrigeration time, but the clarity of the sample is not ideal, and when the refrigeration time is more than 48h, the clarity of the sample is slightly improved, so the refrigeration time is selected to be 48h, and the clarity is optimized on the basis.

Example 4

1. Preference of pH adjustment process

And (3) screening the pH of the refrigerated sample solution, and inspecting the influence of different values on the preparation result when the pH is less than or equal to 7 because the baicalin is unstable under the alkaline condition. Taking 5 equal parts of refrigerated sample liquid, respectively adjusting the pH value to 5.0, 5.5, 6.0, 6.5 and 7.0, respectively adding 50% of sucrose, boiling, filtering, adding water to the amount of the prescription, filling, sterilizing, respectively measuring the contents of baicalin and cichoric acid in the sample before and after sterilization, measuring the pH value of the sample after sterilization, observing the clarity of the sample, and determining the optimal pH value.

TABLE 12 PH adjustment Process Experimental results

From the above results, it can be seen that: adjusting the pH value to 5.0 and 5.5, and still obtaining a small amount of precipitate after sterilization; when the pH value is adjusted to be more than 6.0, the clarity of the sample is gradually improved, but the pH value in the sample is gradually reduced along with the increase of the pH value, and the contents of baicalin and chicoric acid are gradually reduced, so that the pH adjustment process is adjusted to be 6.5 by comprehensive consideration.

Example 5

1.1 sweetener species screening

The sweetening agents commonly used in the market at present mainly comprise sucrose, sodium cyclamate, stevioside and sucralose. The sweetness of the stevioside is 200-300 times that of the cane sugar, and the calorific value of the stevioside is only 1/300 of the cane sugar. The sweetness of the sodium cyclamate is 30-40 times that of the sucrose. The sweetness of the sucralose is about 600 times that of the sucrose, the sweetness is pure, and the sweetness characteristic is very similar to the sweetness quality and the sucrose.

The prescription of syrup in the Chinese pharmacopoeia is as follows: the sucrose content should be not less than 45% (g/mL). Since the sweetness of the sodium cyclamate is low, the experiment carries out comparative research on sucrose, stevioside and sucralose, and the type of the sweetener is determined by taking the taste as an investigation index.

TABLE 13 screening results of sweetener types

The test result shows that: the syrup with only sucrose added had a poor taste; a sweetening agent is added on the basis of adding the cane sugar, so that the taste is poor; the syrup prepared by adding sucrose, stevioside and sucralose has better taste.

1.2 sweetener dosage screening

And screening the using amount of the sweetening agent, and determining the using amount of the sweetening agent by taking the taste as an investigation index.

TABLE 14 screening of sweetener amounts

The test result shows that: when the dosages of the stevioside, the sucralose and the sucrose are respectively 0.2% (g/mL), 0.1% (g/mL) and 45% (g/mL), the taste of the syrup is moderate and relatively good in sweet taste, so that the dosages of the three sweeteners are respectively 45% (g/mL), 0.2% (g/mL) of the stevioside and 0.1% (g/mL) of the sucralose.

Example 6

1.1 screening of essence types

In order to further improve the smell and taste of the preparation, a small amount of essence is added into the preparation. The experiment carries out comparative research on orange essence, strawberry essence, apple essence, lemon essence, chocolate essence and cream essence, and determines the type and the dosage of the essence by taking the taste as an investigation index.

TABLE 15 screening of the flavor types

The test result shows that: the orange essence has fresh taste, can cover certain bitter taste of syrup, and has better taste than other essences, so the orange essence is selected.

1.2 essence dosage screening

On the basis of determining the types of the essences, the dosage of the essences is screened, and the dosage of the essences is determined by taking the mouthfeel as an investigation index.

TABLE 16 results of essence usage

The test result shows that: the orange essence has good taste when the dosage is 0.15% (g/mL).

Example 7

1. Preparation of syrup

500g of dandelion, 188g of isatis root, 125g of corydalis bungeana, 188g of scutellaria baicalensis and scutellaria baicalensis are added after being boiled in water, 10 times of weight of water is added for boiling for three times, 1.5 hours each time, the filtration is carried out, the filtrates are combined, the concentrated solution is concentrated to form clear paste with the relative density of 60-70 ℃ and the thermal test temperature of 1.10-1.15, ethanol is added to ensure that the volume content of the clear paste is 60%, the clear paste is placed for 24 hours, the filtration is carried out, the ethanol is recovered from the filtrate until the relative density is 60-70 ℃ and the thermal test temperature is 1.15-1.20, water is added to 700mL, the cold storage is carried out for 48 hours, the filtration is carried out, the PH value of the filtrate is adjusted to 6.5 by using 10% sodium hydroxide solution, 450g of cane sugar, 2g of stevioside and 1g of trichlorosucrose are added.

3 batches of pilot scale tests were carried out according to the above preparation process, 3 batches of materials were fed, the batch numbers were 150621, 150622, 150623 respectively, and the pilot data of the production process are shown in table 17.

TABLE 17 Mass production data

Injecting: 150621 batches of samples, wherein the extraction batch is 300 times of prescription, and 100 times of prescription extract is used for toxicological tests; 200 times of the prescription extract is used for quality research and stability test. 150622, 150623 batches were used for stability testing.

The baicalin and chicoric acid contents in the three batches of samples (batch numbers 150621, 150622 and 150623) are relatively stable, which shows that the process has good continuous production stability, the process conditions are easy to control and feasible, and the process is suitable for industrial mass production.

Example 8

Stability test

1.1 items under investigation

Table 18 examination item

1.2 accelerated test

A large-scale production sample of the pediatric Pudilan anti-inflammatory syrup (all batches in the sample source) is placed in a constant-temperature constant-humidity box with the temperature of 40 +/-2 ℃ and the relative humidity of 75 +/-5 percent for 6 months. Sampling and detecting once at the end of 0, 1, 2, 3 and 6 months respectively, detecting according to the quality standard for the production of the pediatric Pudilan anti-inflammatory syrup, and comparing the detection result with 0 day, wherein the specific test result is shown in tables 19-21.

TABLE 19 accelerated sample Retention examination results (batch number: 150621)

TABLE 20 accelerated sample Retention examination results (batch number: 150622)

TABLE 21 accelerated sample reservation examination results (batch number: 150623)

As can be seen from tables 19 to 21, the three observed samples were placed in a constant temperature and humidity chamber at 40. + -. 2 ℃ and 75. + -. 5% relative humidity for 6 months under the specified packaging conditions, and compared with 0 day, the properties, identification, relative density, pH, microbial limit, and baicalin content were almost unchanged. The content of chicoric acid is slightly reduced from 0.17mg/mL to about 0.15 mg/mL. But all detection results meet the standard.

1.3 Long term test

And (3) long-term test: a large-scale production sample of the pediatric Pudilan anti-inflammatory syrup (all batches in the sample source) is placed in a constant-temperature and constant-humidity box with the temperature of 25 +/-2 ℃ and the relative humidity of 60 +/-10 percent for 18 months. Sampling is carried out once at the end of 0, 3, 6, 9, 12 and 18 months respectively, detection is carried out according to the clinical quality standard of the pediatric Pudilan anti-inflammatory syrup, the detection result is compared with 0 day, and the specific test result is shown in tables 22-24.

TABLE 22 Long term sample examination results (batch number: 150621)

TABLE 23 Long term sample retention investigation results (batch number: 150622)

Table 24 Long-term sample observation results (batch number: 150623)

As can be seen from tables 22-24, the three observed samples were placed in a constant temperature and humidity chamber at 25 + -2 ℃ and 60% + -10% relative humidity for 18 months under the specified packaging conditions, and compared with 0 day, the properties, identification, relative density, pH value, microbial limit, baicalin content and chicoric acid content were not changed basically, and all the detection results were in accordance with the standard specification.

Example 9

Statistical methods in this example:

data results are expressed as mean ± standard deviation (mean ± SD), and comparisons of differences between groups were statistically significant using Dunnett's test with P <0.05 using the one-way ANOVA analysis method of Graphpad Prism 5 software.

The test drugs and positive drugs in this example:

the effective extract of the infant Pudilan syrup is provided by Jichuan pharmaceutical industry group Limited company, and has the following batch number: 150621. 1g of extract is equivalent to 3.5g of crude drug. The total crude drug amount of the finished product taken by children daily is as follows: 15g crude drug/person/day.

Aspirin enteric tablets (bai xi), Bayer Health Care Manufacturing s.r.l, specification: 100 mg/tablet, batch number: BJ 22790.

Pentoxyverine citrate tablet (Kebiqing), pharmaceutical group Capacity pharmaceutical Co., Ltd, Specification: 25 mg/tablet, 100 tablets/bottle, batch number: 14061722.

1. pudilan syrup dosage design

TABLE 25 rat dose

TABLE 26 mouse doses

TABLE 27 guinea pig dosages

2. Influence of Pudilan blue syrup on acute pharyngitis of rats caused by ammonia water

2.1 Experimental animals

SD rats with SPF grade, weight of 120-: SCXK (Zhe) 2014-0001. The rats are raised in cages and fed with granulated feed at room temperature of 22 +/-2 ℃, humidity of 50% -67%, free drinking water, moderate illumination and adaptive feeding for later use.

2.2 methods

2.2.1 establishment of acute pharyngitis model in rat

SPF SD rat, female and male half, spraying 15% ammonia water with volume fraction on the throat of the rat by using a throat sprayer, spraying 3 pressing each time, 1 time each day in the morning and afternoon, and continuously spraying for 4 days to form an acute pharyngitis model; the appearance state of each group of animals is observed and recorded every day, and the pharyngeal condition of the animals, including the shape, color and the like of mucous membrane, is observed.

2.2.2 animal groups and dosing regimens

72 SD rats, each half of male and female, are randomly divided into 6 groups according to body weight, and each group comprises 12 rats, namely a blank group, a model group, a positive drug group, Pudilan 0.45g/kg, 1.35g/kg and 4.05 g/kg. Rats in each group were gavaged with the corresponding dose of drug starting at d1, and the blank group and the model group were given the same volume of saline 1 time a day for 7 consecutive days. Except for a blank group, the d5-d7 replicates the acute pharyngitis model of the SD rat by adopting ammonia water spray with volume fraction of 15%, and the blank group is sprayed with equal volume of normal saline twice a day.

2.3 results of the experiment

2.3.1 behavioral observations in rats

After spraying ammonia water to rats, most rats have scratching mouth parts, increase oral secretion, increase drinking water, decrease food intake, decrease spontaneous activity and the like. After 4 days of ammonia water molding, the pharynx of the rats in the model group is congested, the rats are dark red and slightly swollen, and the symptoms of the rats in the administration group are relieved to a different degree compared with those in the model group.

2.3.2 blood routine test results

Compared with the blank group, the model group has increased monocyte and neutrophil percentage, decreased lymphocyte and no obvious change in eosinophil percentage. The Pudilan blue syrup with different dosages can reduce the percentage content of monocytes and neutrophils to different degrees and increase the percentage content of lymphocytes, but the percentage content of the Pudilan blue syrup between groups has no obvious difference; the drug had no significant effect on the percentage of eosinophils.

TABLE 28 blood routine test results

2.3.3 expression levels of IL-6 in pharyngeal tissues

After the throat spray by ammonia water, the expression level of IL-6 in pharyngeal mucosa tissues of rats is obviously increased compared with that of a blank group, and the IL-6 content can be obviously reduced by 0.45g/kg of Pudilan syrup.

TABLE 29 expression levels of IL-6 in pharyngeal tissues

Note: in comparison with the blank set, the results,^##P<0.01; comparison with model group<0.05，*P<0.05。

2.3.4 pharyngeal histopathological Observation

The pharyngeal mucosa of a normal rat is covered by stratified squamous epithelium, a thin layer of connective tissue is arranged under the epithelium, mucous glands or mixed glands can be seen, the mucosal epithelial cells are not degenerated and necrotic, and inflammatory cell infiltration is not generated in the epithelium and the hypodermis. In the model group, most pharyngeal wall tissues are slightly congested, pharyngeal mucosal epithelial cells are slightly or moderately degenerated and necrosed, the epithelial cells in the pharynx or the pharyngeal wall are slightly or moderately infiltrated by inflammatory cells, and the inflammatory cell types are mainly neutrophilic granulocytes. The Pudilan blue syrup 4.05g/kg can obviously reduce the pathological change degree of pharyngeal mucosa of rats.

TABLE 30 rat pharyngeal lesion score results (mean + -SD)

Note: in comparison with the blank set, the results,^##P<0.01; in comparison with the set of models,^*P<0.05。

the Pudilan oral liquid has certain intervention effect on the acute pharyngitis of rats according to behavioral observation, blood routine detection results, the expression level of IL-6 in pharyngeal tissues and pharyngeal histopathological observation.

3. Effect of Pudilan blue syrup on citric acid-induced cough in guinea pigs

3.1 Experimental materials

3.1.1 Positive drugs

3.1.2 Experimental animals

Young guinea pigs (8 weeks old), normal grade, half male and female, weight 210-: SCXK (Su) 2012-0008.

The guinea pigs are raised in cages and fed with granulated feed at room temperature of 22 +/-2 ℃, humidity of 50% -67%, free drinking water, moderate illumination and adaptive feeding for later use.

3.2 Experimental methods and dosing regimens

40 young guinea pigs (8 weeks old) with the weight of 210 plus 260g and half male and female are placed in a 500mL observation chamber, 17.5% citric acid is sprayed in at constant pressure for 1min, the cough latency and cough frequency of the guinea pigs within 5min are observed and recorded, and the guinea pigs with the cough latency of less than or equal to 10s or more than or equal to 120s and the cough frequency of less than or equal to 10 times or more than or equal to 50 times within 5min are all removed. Preselected eligible guinea pigs were randomly divided into 5 groups by weight stratification, i.e., model group, tobrex group, low, medium, and high dose groups (0.39g/kg, 1.16g/kg, 3.49g/kg) of pediatric Pudilan syrup, and 6 animals per group. Each group was administered by continuous gavage for 7 days, once a day, and model groups were fed with equal volumes of distilled water. 1h after the last administration, the guinea pigs with cough are induced according to the screening conditions, and the cough incubation period of the guinea pigs and the number of coughs within 5min are observed and recorded.

3.3 results

Compared with a model group, the low, medium and high doses of the pediatric Pudilan syrup can prolong the cough latency of cough-inducing guinea pigs to different degrees (P <0.05, P <0.01), and the best dose is 3.49 g/kg; in addition, the cough frequency of the cough-inducing guinea pigs can be reduced by the pediatric Pudilan syrup in each dosage group (P < 0.001).

TABLE 31 Effect of pediatric Pudilan syrup on citric acid induced cough in Guinea pigs

Note: p <0.05, P <0.01, P <0.001, compared to model groups.

The cough relieving effect of the Pudilan blue syrup is investigated through a guinea pig cough inducing experiment, in the experiment, each dosage group of the syrup can prolong the cough latent period to different degrees, and the cough frequency of the young guinea pig caused by citric acid is reduced, so that the infant Pudilan blue syrup has an obvious cough relieving effect.

4. Effect of Pudilan blue syrup on Ammonia induced cough in mice

4.1 Experimental materials

4.1.1 Positive drugs

Pentoxyverine citrate tablet (Kebiqing), national drug group Capacity pharmaceutical Co., Ltd, Specification: 25 mg/tablet, 100 tablets/bottle, batch number: 14042721.

4.1.2 Experimental animals

ICR mouse, clean grade, male and female half, weight 18-22g, Yangzhou university center of comparative medicine, license number: SCXK (Su) 2012-0004.

Mice are raised in cages and fed with granulated feed, the room temperature is 22 +/-2 ℃, the humidity is 50% -67%, water is freely drunk, the illumination is moderate, and the mice are fed adaptively for later use.

4.2 Experimental methods and dosing regimens

60 ICR mice 18-22g, each male and female half, were randomly divided into 5 groups according to body weight, namely model group, tubibenz group, Pudilan syrup low, medium and high dose groups (0.65g/kg, 1.95g/kg, 5.85 g/kg). Each group was administered by continuous gavage for 7 days, once a day, and model group mice were filled with equal volume of distilled water. 1h after the last administration, the mice were placed in an inverted beaker with a 2L volume and a cotton ball placed inside, timing was started when 0.5mL of 25% concentrated ammonia water was added dropwise to the cotton ball, and the cough latency and the number of coughs within 3min of the mice were observed and recorded.

Compared with a model group, the low, medium and high doses of the Pudilan syrup can obviously reduce the cough frequency (P <0.05, P <0.01 and P <0.001) of mice within 3min, and the cough latency (P <0.05) can be obviously prolonged at the dose of 5.85 g/kg.

TABLE 32 Effect of pediatric Pudilan syrup on Ammonia induced cough in mice

Note: each group was compared to the model group, P <0.05, P <0.01, P < 0.001.

The cough relieving effect of the Pudilan syrup is examined through a mouse cough inducing experiment. Research results show that in an ammonia water-induced mouse cough experiment, 5.85g/kg of syrup can obviously reduce the mouse cough frequency and prolong the cough latency period, and the Pudilan syrup has an obvious cough relieving effect.

5. Effect of Pudilan blue syrup on mouse ear swelling caused by xylene

5.1 Experimental animals

5.2 Experimental methods and dosing regimens

ICR mice, which are respectively half male and female, are 72 mice, and after being adaptively raised under normal conditions, the ICR mice are randomly divided into 6 groups according to the body weight, namely a blank group, a model group, an aspirin group and a Pudilan syrup low, medium and high dose group (0.65g/kg, 1.95g/kg and 5.85 g/kg). The mice in each group are respectively administrated with the corresponding medicine by gavage, the administration volume is 0.1mL/10g, and the blank group and the model group are administrated with distilled water with the same volume for 1 time per day and 7 days continuously. 1h after the last administration, except for a blank group, xylene is uniformly coated on the right ear of each group of mice by a micro-injector, each group of mice is 30 mu m/mouse, after 30min of inflammation, the thickness of the double ears of the mice is measured by a thickness gauge, and the difference between the left ear and the right ear is calculated. And (4) taking an ear disk, weighing the ear disk with the diameter of 0.8cm, and calculating the ear weight difference of the ears.

5.3 Effect of Pudilan on mouse ear swelling degree caused by paraxylene

The ear thickness and the ear weight of the model group mice are obviously increased compared with the blank group (P <0.001), the ear thickness of the mice is obviously reduced by 1.95g/kg and 5.85g/kg of Pudilan syrup (P <0.001), and the ear weight of the mice can also be obviously reduced by 5.85g/kg of group (P < 0.05).

TABLE 33 influence of Pudilan on the degree of ear swelling in mice ear swelling model by paraxylene

Note:^###P<0.001 compared to blank; p<0.05，**P<0.01 was compared to the model group.

In the experiment, a xylene-induced mouse ear swelling model is adopted to investigate the anti-inflammatory effect of the Pudilan syrup. Research results show that the Pudilan can obviously relieve the ear swelling degree of a model mouse, and the Pudilan syrup has a certain anti-inflammatory effect.

6. Influence of Pudilan blue syrup on permeability of capillary vessels in abdominal cavity of mouse

6.1 Experimental animals

6.2 Experimental methods and dosing regimens

60 ICR mice 18-22g, each male and female half, according to the weight random stratification into 5 groups, namely model group, aspirin group (400mg/kg), Pudilan syrup low, medium, high dose group (0.65g/kg, 1.95g/kg, 5.85g/kg), the administration volume of 0.1mL/10 g. Each group was administered by continuous gavage for 7 days, once a day, and model group mice were filled with equal volume of distilled water.

1h after the last administration, tail vein is injected with 0.1mL/10g of 2% Evans blue physiological saline solution, then 0.2 mL/only 0.6% acetic acid physiological saline solution is injected into abdominal cavity, the abdominal cavity is killed after 30min, the abdominal cavity is opened, 6mL of physiological saline is used for washing the abdominal cavity, washing liquid is sucked out, supernatant (3000r/min) is obtained by centrifugation, and the OD value of optical density is measured at 590 nm.

6.3 results of the experiment

Compared with a model group, the Pudilan blue syrup of 5.85g/kg and 1.95g/kg can inhibit the increase of the capillary permeability of the mice caused by acetic acid, and the 5.85g/kg dosage group has a significant difference (P <0.05) compared with the model group.

TABLE 34 Effect of Pudilan syrup on mouse capillary Permeability

Note: each group was compared to the model group,. P < 0.05.

The inhibition effect of the Pudilan blue syrup on the increase of the capillary permeability of the mice caused by acetic acid is examined through a mouse peritoneal capillary permeability experiment. Research results show that Pudilan blue syrup 1.95g/kg and 5.85g/kg can significantly reduce capillary permeability of mice, and the Pudilan blue syrup can play an anti-inflammatory role by inhibiting capillary permeability and reducing local aseptic inflammatory exudation.

Pharmacodynamic test research results show that the pediatric Pudilan anti-inflammatory syrup has a certain intervention effect on acute pharyngitis of rats; the incubation period of guinea pig cough caused by citric acid can be prolonged, and the cough frequency of the guinea pig is reduced; can also prolong the latent period of mouse cough induced by ammonia water, and has remarkable cough relieving effect; the mouse ear swelling caused by dimethylbenzene is obviously improved, and the mouse ear swelling caused by dimethylbenzene has a certain anti-inflammatory effect; can inhibit capillary permeability, and reduce topical aseptic inflammation exudation to exert antiinflammatory effect.

The pediatric Pudilan anti-inflammatory syrup has certain anti-inflammatory and cough-relieving effects, and is suitable for treating infantile epidemic parotitis, acute pharyngitis and acute tonsillitis. The formula has reasonable, safe and effective compatibility, and stable and controllable process and quality.

Claims

1. A preparation method of Pudilan extract with anti-inflammatory effect comprises the following steps:

1) water extraction: 3.0-5.0 parts of dandelion, 1.0-2.0 parts of isatis root, 0.5-1.5 parts of bunge corydalis herb and 1.0-2.0 parts of scutellaria baicalensis are extracted by water, wherein after the dandelion, the isatis root and the bunge corydalis herb are boiled in water, the scutellaria baicalensis is put into the water and extracted by the water to obtain water extract; in the water extraction step, the weight of water is 10 times of that of the decocted raw materials, the decoction is carried out for 3 times, each time lasts for 1.5 hours, and the decoction liquids of the times are combined;

2) alcohol precipitation: concentrating the water extractive solution to relative density of 60-70 deg.C, measuring with heat at 1.10-1.15, adding anhydrous ethanol to make ethanol volume content in the concentrated water extractive solution be 60%, standing for 24 hr, filtering to remove insoluble substances, and removing ethanol from the filtrate to obtain extract.

2. The method for preparing Pudilan extract according to claim 1, wherein the raw material drugs comprise 4 parts by weight of dandelion, 1.5 parts by weight of radix isatidis, 1 part by weight of corydalis bungeana and 1.5 parts by weight of scutellaria baicalensis.

3. An extract of Pudilan with anti-inflammatory effect, which is prepared by the method of any one of claims 1 to 2.

4. A Pudilan anti-inflammatory drug, which consists of the extract of claim 3 as an active ingredient and pharmaceutically acceptable excipients.

5. The Pudilan anti-inflammatory drug of claim 4 which is a syrup, said excipient comprising a sweetener.

6. The Pudilan anti-inflammatory drug according to claim 5, wherein the sweetener is one or more of sucrose, steviol glycosides and sucralose.

7. The Pudilan anti-inflammatory drug of claim 6, wherein the sweetener is sucrose 45% (g/mL), stevioside 0.2% (g/mL) and sucralose 0.1% (g/mL).

8. The Pudilan anti-inflammatory agent as claimed in any one of claims 4 to 7, wherein the adjuvant comprises perfume.

9. The Pudilan anti-inflammatory agent of claim 8, wherein the flavoring is orange flavoring.

10. The Pudilan anti-inflammatory agent of claim 9, wherein the orange flavor is present in an amount of 0.15% (g/mL).

11. The Pudilan antiphlogistic syrup is prepared from 500g of raw material medicines of dandelion, 188g of isatis root, 125g of corydalis bungeana and 188g of scutellaria baicalensis, and the preparation method comprises the following steps:

1) water extraction: boiling herba Taraxaci, radix Isatidis and herba corydalis Bungeanae in water, adding Scutellariae radix, and extracting with water to obtain water extractive solution; in the water extraction step, the weight of water is 10 times of that of the decocted raw materials, the decoction is carried out for 3 times, each time lasts for 1.5 hours, and the decoction liquids of the times are combined;

2) alcohol precipitation: concentrating the extractive solution to obtain fluid extract with relative density of 60-70 deg.C and thermal measurement of 1.10-1.15, adding anhydrous ethanol to make ethanol volume content in the concentrated water extractive solution be 60%, standing for 24 hr, filtering, and removing ethanol from the supernatant to obtain fluid extract with relative density of 60-70 deg.C and thermal measurement of 1.15-1.20;

3) adding water to the extract to 700mL, refrigerating for 48h, filtering, adjusting p H value of the supernatant to 6.5 with sodium hydroxide solution, adding sucrose 450g, stevioside 2g and sucralose 1g, boiling, filtering, adding 1.5g of orange essence, and adding water to 1000 mL.

12. The use of the extract according to claim 3 in the manufacture of a medicament for the treatment of mumps in children, acute pharyngitis or acute tonsillitis.