A kind of medicine composition for anti-inflammatory, analgesic and throat clearing and its preparation method

Abstract

The invention discloses a pharmaceutical composition for anti-inflammatory, analgesic and throat clearing and a preparation method thereof, and relates to the field of biomedicine technology. The pharmaceutical composition for anti-inflammatory, analgesic and throat clearing comprises Radix Isatidis polypeptide, concentrated powder of buffalo horn, wild chrysanthemum and schizonepeta spicate. The Radix Isatidis polypeptide GLAASHHLGGK prepared by the invention has a DPPH free radical scavenging ability equivalent to that of Vc and a higher ABTS free radical scavenging ability; the pharmaceutical composition for anti-inflammatory, analgesic and throat clearing containing Radix Isatidis polypeptide can improve the proliferation ability of spleen lymphocytes of immunocompromised mice and improve the NK cell activity of immunocompromised mice, can effectively prevent and improve diseases, enhance disease resistance, improve the body's immunity, and can be used as a drug for improving the body's immunity. The present application discovered the active polypeptide of Radix Isatidis, which filled the gap in the market and expanded the prospects for the development and application of Radix Isatidis.

Description

Technical Field

[0001] The invention relates to the field of biomedicine, and in particular to a pharmaceutical composition for reducing inflammation, relieving pain, and clearing the throat, and a preparation method thereof. Background Art

[0002] Sore throat is one of the most common symptoms. It often occurs in the cold season of the year. Colds, tonsillitis, sinusitis, whooping cough, pharyngitis, viral infections and even myocardial infarction can cause sore throat. Sore throat is the main symptom of oropharyngeal and laryngopharyngeal lesions. It is characterized by redness, swelling and pain in the throat and discomfort in swallowing. It is also called laryngeal paralysis. It is seen in acute tonsillitis, acute pharyngitis and simple laryngitis and tonsillar abscess in Western medicine. Most acute sore throats will disappear automatically within a few days to weeks. However, if the pain persists or worsens within a few days, you need to see a doctor. Because if it is not treated for a long time, the disease may cause rheumatic fever and harm the heart and kidneys. The current treatment drugs are mainly anti-inflammatory drugs and antibiotics. Anti-inflammatory drugs treat the symptoms but not the root cause, and antibiotics are drugs that are restricted for use by the state. Therefore, it is of great practical value to study and develop a safe and effective Chinese medicine for anti-inflammatory, analgesic and throat clearing.

[0003] In the prior art, the traditional Chinese medicines commonly used for anti-inflammatory, analgesic and throat clearing include buffalo horn, wild chrysanthemum, isatis root, schizonepeta spicate and the like.

[0004] For example, the prior art CN 109925338 A discloses a throat pain tablet and a preparation method thereof. The provided throat pain tablet is composed of the following raw materials in parts by weight: 90-120 parts of throat pain tablet thick paste, 90-110 parts of buffalo horn concentrated powder, 58.7-72.2 parts of sucrose powder, 10-20 parts of microcrystalline cellulose, 2-6 parts of limonene, 8-18 parts of β-cyclodextrin, 1.5-3.3 parts of magnesium stearate, 100-130 parts of talc, 90-110 parts of sucrose powder for coating, 0.1-1.5 parts of gelatin, 0.1-1.5 parts of mung bean starch, 0.1-1.2 parts of insect wax and 0-0.5 parts of brilliant blue pigment. Throat Soreness Tablets have the effects of clearing heat, detoxifying, anti-inflammatory and relieving sore throat. They also have the advantages of low hygroscopicity, good stability, not easy to be oxidized and deteriorate, easy to store; high bioavailability, and easy to be absorbed by the human body.

[0005] For example, the prior art CN106619811B discloses an antiviral Chinese medicine compound preparation, wherein the Chinese medicine compound composition is made of 150g wild chrysanthemum, 200g isatis root, 150g schizonepeta spicate, and 20g buffalo horn concentrated powder, the carrier material of the solid dispersion is composed of xylitol and povidone in a weight ratio of 1:2, and the povidone is a mixture of povidone K30 and povidone K90 in a weight ratio of 1:1; the weight ratio between the Chinese medicine compound composition and the solid dispersion carrier material is 1:5; the preparation method of the Chinese medicine compound composition is: soaking 150g wild chrysanthemum, 200g isatis root, and 150g schizonepeta spicate in 95% ethanol for 24 hours, heating and reflux 3 times, each time for 2-3 hours, reducing pressure to recover, drying to obtain an extract, and mixing the extract with 20g buffalo horn concentrated powder to obtain.

[0006] For example, the prior art CN 104435109 B discloses a method for preparing a Houtongling granule composition, and the steps are as follows:

[0007] Step 1: 150 g of wild chrysanthemum, 200 g of isatis root and 150 g of schizonepeta spicate were extracted with 3-10 times the weight of 90-100% (v / v) ethanol.

[0008] Take 1-3 times, combine and concentrate the extracts;

[0009] Step 2, adjusting the pH value of the concentrate to 9, adding ethanol, standing, and filtering;

[0010] Step 3, the filtrate is passed through a D392 resin column, the effluent is collected, the pH value is adjusted to 6.5-7.0, and concentrated to obtain an extract;

[0011] Step 4: After the extract and 20 g of buffalo horn concentrated powder are mixed, ethanol, starch, sucrose powder and dextrin are added as active ingredients of the medicine, and granules are prepared according to conventional pharmaceutical techniques.

[0012] South isatis root is the dried root of the plant Malan from the Acanthaceae family, while isatis root is the dried root of the plant Songlan from the Cruciferae family, also called northern isatis root. Both have the effects of clearing away heat and detoxifying, cooling blood and relieving throat, and can be used to treat sore throat caused by wind-heat cold. The above-mentioned prior art uses isatis root, not south isatis root. However, the material basis of isatis root antiviral is unknown, and the main clinical application dosage form is granules. This preparation has a large dosage, and the quality control index is unclear, which limits the secondary development of the product. There are a lot of reports on the research of the effective parts of isatis root, such as organic acids, amino acids, total alkaloids, epigoitrin, nucleosides, polysaccharides, polypeptides, etc., all of which have good antiviral effects and are considered to be the active ingredients of isatis root. The components of isatis root are complex and diverse, and the understanding of its heat-clearing and detoxifying material basis is not unified. However, few people have studied the active ingredients of south isatis root, so this application will try to develop and study the polypeptides of south isatis root to fill the gap in the market. Summary of the invention

[0013] To achieve the above object, the present invention provides a pharmaceutical composition for relieving inflammation, relieving pain and clearing throat, which comprises Radix Isatidis polypeptide, concentrated buffalo horn powder, wild chrysanthemum and Nepeta spicate.

[0014] In certain embodiments, the amino acid sequence of the Radix Isatidis polypeptide is as described in SEQ ID NO:1.

[0015] In certain embodiments, the anti-inflammatory, analgesic and throat-clearing pharmaceutical composition comprises, by weight, 0.5-5 parts of Radix Isatidis polypeptide, 10-30 parts of buffalo horn concentrated powder, 100-300 parts of wild chrysanthemum, and 100-300 parts of Nepeta spicate.

[0016] In certain embodiments, the anti-inflammatory, analgesic and throat-clearing pharmaceutical composition comprises 2 parts of Radix Isatidis polypeptide, 20 parts of buffalo horn concentrated powder, 200 parts of wild chrysanthemum, and 200 parts of Nepeta spicate.

[0017] In certain embodiments, an auxiliary material is further included.

[0018] In certain embodiments, the auxiliary material is selected from one or more of corn starch, magnesium stearate, sucrose, gelatin, talc, and insect wax.

[0019] The present invention also provides a method for preparing a pharmaceutical composition for reducing inflammation, relieving pain and clearing the throat, comprising the following steps:

[0020] (1): Wash the southern isatis root, dry it naturally in the shade, and then crush it, add 20 times the mass of distilled water, mix and stir to dissolve, ultrasonicate it under 40kHz ultrasonic conditions for 30 minutes, and then centrifuge it at 4000rpm for 15 minutes to 30 minutes, discard the supernatant, and keep the precipitate;

[0021] (2): The precipitate prepared in step 1 is dissolved in distilled water to form a 40% solution by mass, and 1 / 30 of the mass of a composite enzyme is added for enzymatic hydrolysis, wherein the mass ratio of trypsin (200,000 U / g), cellulase (100,000 U / g), and pectinase (100,000 U / g) is 1:1:1, and the enzymatic hydrolysis is carried out at 40° C. for 4 h. The solution is inactivated at 90° C. for 15 min, and centrifuged at 4000 rpm for 15 min. The supernatant obtained is the southern isatis root enzymatic hydrolyzate;

[0022] (3) The isatis indigotica hydrolysate was ultrafiltered using a Biomax modified polyethersulfone composite membrane package with a membrane molecular cutoff of 3 kDa to separate and purify two polypeptide solutions with molecular weights of A1 (>3 kDa) and A2 (<3 kDa);

[0023] (4) Component A2 was separated by Sephadex G-20 to obtain A2-1, A2-2, A2-3, and A2-4

[0024] Four components;

[0025] (5) Separating the A2-3 component by Sephadex G-15 to obtain four components: A2-3-1, A2-3-2, A2-3-3, and A2-3-4;

[0026] (6) centrifuging the A2-3-4 fraction, taking the supernatant, and freeze-drying it to obtain a Radix Isatidis polypeptide, the amino acid sequence of which is shown in SEQ ID NO: 1;

[0027] (7) The Radix Isatidis polypeptide shown in SEQ ID NO: 1, concentrated powder of buffalo horn, wild chrysanthemum, schizonepeta spicate, and auxiliary materials are mixed in the above-mentioned proportions to prepare a pharmaceutical composition containing the Radix Isatidis polypeptide for anti-inflammatory, analgesic, and throat clearing.

[0028] Finally, the present invention provides the use of the above-mentioned anti-inflammatory, analgesic and throat-clearing pharmaceutical composition in the preparation of medicines for improving body immunity or anti-inflammatory, analgesic and throat-clearing.

[0029] The technical solution of the present invention has the following effects:

[0030] 1. The Radix Isatidis polypeptide GLAASHHLGGK (SEQ ID NO: 1) prepared by the present invention has a DPPH free radical scavenging ability comparable to that of Vc and a higher ABTS free radical scavenging ability, that is, the Radix Isatidis polypeptide has a strong antioxidant activity;

[0031] 2. The anti-inflammatory, analgesic and throat-clearing pharmaceutical composition containing the southern isatis root polypeptide prepared by the present invention can improve the proliferation ability of spleen lymphocytes of immunocompromised mice and improve the NK cell activity of immunocompromised mice. The anti-inflammatory, analgesic and throat-clearing pharmaceutical composition can effectively prevent and improve diseases, enhance disease resistance, and improve the body's immunity. Therefore, the anti-inflammatory, analgesic and throat-clearing pharmaceutical composition containing the southern isatis root polypeptide can be used as a drug for improving the body's immunity.

[0032] 3. This application discovered the active polypeptides of southern isatis root, filling the gap in the market and expanding the prospects for the development and application of southern isatis root. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] Figure 1 The results of DPPH free radical scavenging ability and ABTS free radical scavenging ability of the four components A2-1, A2-2, A2-3, and A2-4;

[0034] Figure 2 The results of DPPH free radical scavenging ability and ABTS free radical scavenging ability of the four components A2-3-1, A2-3-2, A2-3-3, and A2-3-4;

[0035] Figure 3 This is the secondary mass spectrum of the Radix Isatidis polypeptide GLAASHHLGGK with a clear amino acid sequence;

[0036] Figure 4 The effect of the anti-inflammatory, analgesic and throat-clearing pharmaceutical composition containing GLAASHHLGGK Radix Isatidis polypeptide on the proliferation capacity of mouse spleen lymphocytes;

[0037] Figure 5 The invention discloses an effect of an anti-inflammatory, analgesic and throat-clearing pharmaceutical composition containing GLAASHHLGGK Radix Isatidis polypeptide on the activity of NK cells in mice. DETAILED DESCRIPTION

[0038] The methods and applications of the present invention have been described through preferred embodiments. Relevant personnel can obviously modify or appropriately change and combine the methods and applications herein without departing from the content, spirit and scope of the present invention to implement and apply the technology of the present invention.

[0039] Example 1: Preparation and identification of Isatis indigotica polypeptides

[0040] (1): Wash the southern isatis root, dry it naturally in the shade, and then crush it. Add 20 times the mass of distilled water and stir to dissolve it. Ultrasonicate it under 40kHz ultrasonic conditions for 30 minutes, then centrifuge it at 4000rpm for 15 minutes to 30 minutes, discard the supernatant, and keep the precipitate.

[0041] (2): The precipitate prepared in step 1 is dissolved in distilled water to form a solution with a mass fraction of 40%, and 1 / 30 of the mass of a composite enzyme is added for enzymatic hydrolysis, wherein the mass ratio of trypsin (200,000 U / g), cellulase (100,000 U / g), and pectinase (100,000 U / g) is 1:1:1. The enzymatic hydrolysis is carried out at 40°C for 4 hours, the solution is inactivated at 90°C for 15 minutes, and centrifuged at 4000 rpm for 15 minutes. The supernatant obtained is the enzymatic hydrolyzate of Radix Isatidis indigotica.

[0042] (3): The southern isatis root hydrolyzate in step 2 was ultrafiltered using a Biomax modified polyethersulfone composite membrane package with a membrane molecular cutoff of 3 kDa to separate and purify two molecular weight polypeptide solutions, A1 (>3 kDa) and A2 (<3 kDa). The A2 component was collected and desalted using a dialysis bag (500 Da), concentrated by rotary evaporation, freeze-dried, and stored in a -20°C refrigerator for later use.

[0043] (4): The A2 component was separated and purified by Sephadex G-20, wherein the separation range of Sephadex G-20 was approximately 0.3 kDa to 5 kDa, and four components A2-1, A2-2, A2-3, and A2-4 were separated. The obtained components were concentrated by rotary evaporation and vacuum freeze-dried, and the freeze-dried polypeptide powder was collected and prepared into a certain concentration for experimental study on DPPH free radical scavenging ability and ABTS free radical scavenging ability;

[0044] DPPH free radical scavenging ability test:

[0045] Add an equal amount of sterile distilled water to prepare 0.6 mg / mL Radix Isatidis polypeptide in Group A2-1, Group A2-2, Group A2-3, and Group A2-4. Take 1 mL of Radix Isatidis polypeptide sample solutions from different groups and add them to 2 mL of DPPH (0.1 mM, methanol solution). After mixing, place them at room temperature in the dark for 30 minutes, and measure the absorbance at 517 nm. Distilled water was used as the blank group, ascorbic acid (Vc) was used as the positive control, and all samples were measured in three groups in parallel and the average value was calculated. The scavenging ability is represented by the scavenging rate, and the calculation formula is as follows:

[0046] Clearance rate (parts) = [A0 - (Ai - A1)] / A0 × 100 parts

[0047] A0——absorbance of blank group (1 mL of distilled water instead of peptide sample);

[0048] Ai——absorbance of sample solution;

[0049] A1——Absorbance of 2mL methanol instead of DPPH.

[0050] ABTS free radical scavenging ability test:

[0051] Preparation of ABTS free radical stock solution: Mix 10mL of ABTS solution with a concentration of 7.4mmol / L and 10mL of K2S2O8 solution with a concentration of 2.6mmol / L, and oxidize at room temperature in the dark for 12h to obtain a stock solution. Dilute the ABTS stock solution with PBS (0.2M, pH=7.4) before use, and make its absorbance value at 734nm 0.80±0.02. Add an equal amount of sterile distilled water to prepare 0.6mg / mL southern isatis root polypeptide in group A2-1, group A2-2, group A2-3, and group A2-4, take 2mL of ABTS solution, add different groups of southern isatis root polypeptide sample solutions, mix, let stand for 6min at room temperature, and measure the absorbance at 734nm. Use ascorbic acid (Vc) as a positive control, and measure three groups of all samples in parallel to calculate the average value. The ABTS scavenging ability is expressed by the clearance rate, and the calculation formula is as follows:

[0052] Clearance rate (parts) = [A0 - (Ai - A1)] / A0 × 100 parts

[0053] Wherein, A0——blank control (1 mL of distilled water instead of peptide sample);

[0054] Ai——absorbance of sample solution;

[0055] A1 - sample control (2 mL 0.2 M buffer instead of ABTS).

[0056] like Figure 1 As shown, the A2-3 group had the highest scavenging rate of Radix Isatidis polypeptide on DPPH and ABTS free radicals, which was comparable to the effect of Vc and significantly different from the effects of other groups (P<0.01).

[0057] Step 5: Separate and purify the A2-3 component by Sephadex G-15, wherein the separation range of Sephadex G-15 is approximately between 0.7 kDa and 1.5 kDa; concentrate the obtained A2-3-1, A2-3-2, A2-3-3, and A2-3-4 components by rotary evaporation, and freeze-dry them in a vacuum. Collect the freeze-dried polypeptide powder and prepare it into a certain concentration for experimental study on DPPH free radical scavenging ability and ABTS free radical scavenging ability. The determination experimental method is the same as that in step 4.

[0058] like Figure 2 As shown, the A2-3-4 group had the highest scavenging rate of Radix Isatidis polypeptide on DPPH and ABTS free radicals, which was comparable to the effect of Vc and significantly different from the effects of other groups (P<0.01).

[0059] Step 6: Purify the active peptides of group A2-3-4 by RP-HPLC, and obtain the peak of the product separated and purified by RP-HPLC, the purification conditions are: column temperature 25°C, flow rate 0.6mL / min, injection volume 4μL; then, identify the active peptides by LC-ESI-MS / MS and obtain the southern isatis root polypeptide with antioxidant capacity of the present invention ( Figure 3 ), the clear secondary mass spectrum of the southern isatis polypeptide GLAASHHLGGK, the identification conditions are: mass scanning range 1200m / z, drying gas flow rate 5L / min, drying gas temperature 150℃.

[0060] Example 2 Preparation and testing of anti-inflammatory, analgesic and throat-clearing pharmaceutical composition

[0061] The anti-inflammatory, analgesic and throat-clearing pharmaceutical composition is prepared by weight: 2 parts of southern isatis root polypeptide, 20 parts of buffalo horn concentrated powder, 200 parts of wild chrysanthemum, and 200 parts of schizonepeta spicate.

[0062] (1) Immunity test:

[0063] After SPF-grade C57BL / 6 mice were raised stably for a period of time, the experimental mice were randomly divided into a blank control group, a model control group, an experimental group 1 and an experimental group 2 according to their body weight.

[0064] Blank control group: intraperitoneal injection of normal saline;

[0065] Experimental group 1: Radix Isatidis polypeptide GLAASHHLGGK group;

[0066] Experimental group 2: a pharmaceutical composition group containing the Radix Isatidis polypeptide GLAASHHLGGK for anti-inflammatory, analgesic and throat clearing;

[0067] The mice in the model control group, experimental group 1 and experimental group 2 were intraperitoneally injected with 80 mg / kg cyclophosphamide for 3 consecutive days from the day of grouping. Thereafter, the model was reinforced with injection once a week. The blank control group was intraperitoneally injected with an equal amount of saline during the same period. The modeling period was 30 days.

[0068] During the modeling period, the experimental group 1 was gavaged with 0.4 mg / kg of polypeptide, and the experimental group 2 was gavaged with 40 mg / kg of the anti-inflammatory, analgesic and throat-clearing pharmaceutical composition containing southern isatis root multi-GLAASHHLGGK, once a day for 30 consecutive days; the blank control group and the model control group were gavaged with the corresponding volume of pure water every day. Subsequently, the above mice were used for ConA (concanavalin A)-induced mouse spleen lymphocyte transformation and mouse spleen NK cell activity assay tests.

[0069] See also Figure 4-Figure 5Compared with the model control group, the Radix Isatidis polypeptide GLAASHHLGGK group and the modulated milk powder group containing Radix Isatidis polypeptide GLAASHHLGGK could significantly improve the proliferation ability of spleen lymphocytes and NK cell activity of mice (P<0.01).

[0070] (2) Pharyngitis test:

[0071] Experimental methods: The preparation method of the animal model refers to Tan Yanhui et al., Study on the protective effect of the water extract of Dong medicine Xuemabian on rats with pyridine-induced acute non-infectious pharyngitis, Tianjin Journal of Traditional Chinese Medicine, Vol. 36, No. 2, February 2019, pp. 176-180;

[0072] SD rats were randomly divided into a normal control group (oral administration of normal saline, pharyngeal swab of normal saline), a model control group (oral administration of normal saline; pharyngeal swab of 10% pyridine), an experimental group 1: pyridine + southern isatis polypeptide oral administration group (oral administration of 0.4 mg / kg, pharyngeal swab of 10% pyridine), an experimental group 2: pyridine + southern isatis polypeptide containing anti-inflammatory, analgesic and throat-clearing pharmaceutical composition (oral administration of 40 mg / kg, pharyngeal swab of 10% pyridine); pyridine + dexamethasone group (oral administration of dexamethasone 4 mg / kg, pharyngeal swab of 10% pyridine). Rats were pretreated with drugs for 5 consecutive days, once in the morning and evening every day. One hour after the last administration, the rats were anesthetized by intraperitoneal injection of sodium pentobarbital (dose 60 mg / kg, concentration 2%). About 5 minutes later, EB (dose 30 mg / kg, concentration 5 mg / mL) was injected into the tail vein of the rats. After 10 minutes, the rat's mouth was opened with hemostats and forceps, the rat's tongue was slightly lifted outward and pulled to one side, 130 μL of 10% pyridine solution was absorbed with a cotton swab, and it was inserted into the rat's pharynx until resistance was felt, then slightly pulled back, and stopped and rotated for 5 seconds, for 3 consecutive times. After 1 hour of modeling, the rat was killed, photographed and collected. The EB-formamide standard curve was prepared and the EB permeation content in the pharyngeal tissue was determined with reference to the above literature, and the results are shown in Table 1.

[0073] Table 1. Experimental results

[0074] Grouping dose Number of animals EB exudate (μg / g tissue) Normal control group Normal saline 5 0.35±0.05 Model control group Normal saline 5 0.69±0.09 Dexamethasone group 4mg / kg 5 0.37±0.06 Experimental Group 1 0.4mg / kg 5 0.37±0.05 Experimental Group 2 40mg / kg 5 0.35±0.07

[0075] As shown in the results in Table 1, compared with the model control group, the experimental groups 1 and 2 had the ability to treat the pathological damage of the pharyngeal tissue in rats with acute pharyngitis (P<0.01).

[0076] The above-mentioned embodiments only express several implementation methods of the present application, and the descriptions thereof are relatively specific and detailed, but they cannot be understood as limiting the scope of the invention patent. It should be pointed out that, for a person of ordinary skill in the art, several variations and improvements can be made without departing from the concept of the present application, and these all belong to the protection scope of the present application. Therefore, the protection scope of the patent of the present application shall be subject to the attached claims.

Claims

1. A pharmaceutical composition for reducing inflammation, relieving pain and clearing the throat, characterized in that: The composition comprises Radix Isatidis indigotica polypeptide, concentrated buffalo horn powder, wild chrysanthemum and Nepeta spicate; The amino acid sequence of the Radix Isatidis polypeptide is shown in SEQ ID NO: 1; The anti-inflammatory, analgesic and throat-clearing pharmaceutical composition comprises, by weight, 0.5 to 5 parts of Radix Isatidis polypeptide, 10 to 30 parts of concentrated buffalo horn powder, 100 to 300 parts of wild chrysanthemum, and 100 to 300 parts of Nepeta spicate.

2. The anti-inflammatory, analgesic and throat-clearing pharmaceutical composition according to claim 1, characterized in that: The anti-inflammatory, analgesic and throat-clearing pharmaceutical composition comprises 2 parts of Radix Isatidis polypeptide, 20 parts of buffalo horn concentrated powder, 200 parts of wild chrysanthemum and 200 parts of Nepeta spicate.

3. The anti-inflammatory, analgesic and throat-clearing pharmaceutical composition according to claim 1, characterized in that: Also includes excipients.

4. The anti-inflammatory, analgesic and throat-clearing pharmaceutical composition according to claim 3, characterized in that: The auxiliary material is selected from one or more of corn starch, magnesium stearate, sucrose, gelatin, talcum powder and insect wax.

5. A method for preparing a pharmaceutical composition for reducing inflammation, relieving pain and clearing the throat, comprising the following steps: (1): Wash the southern isatis root, dry it naturally in the shade, and then crush it, add 20 times the mass of distilled water, mix and stir to dissolve, ultrasonicate it under 40kHz ultrasonic conditions for 30 minutes, and then centrifuge it at 4000rpm for 15 minutes to 30 minutes, discard the supernatant, and keep the precipitate; (2): The precipitate prepared in step 1 is dissolved in distilled water to form a 40% solution by mass, and 1 / 30 of the mass of a composite enzyme is added for enzymatic hydrolysis, wherein the mass ratio of trypsin, cellulase and pectinase in the composite enzyme is 1:1:1, and the enzymatic hydrolysis is carried out at 40° C. for 4 h, the solution is inactivated at 90° C. for 15 min, and centrifuged at 4000 rpm for 15 min. The supernatant obtained is the southern isatis root enzymatic hydrolyzate; The concentration of the trypsin is 200,000 U / g; The concentration of the cellulase is 100,000 U / g; The concentration of the pectinase is 100,000 U / g; (3) The isatis indigotica hydrolysate was ultrafiltered using a Biomax modified polyethersulfone composite membrane package with a membrane molecular cutoff of 3 kDa to separate and purify two molecular weight polypeptide solutions, A1 and A2; The molecular weight of A1 is greater than 3 kDa; the molecular weight of A2 is less than 3 kDa; (4) Separating the A2 component by Sephadex G-20 to obtain four components: A2-1, A2-2, A2-3, and A2-4; (5) Separating the A2-3 component by Sephadex G-15 to obtain four components: A2-3-1, A2-3-2, A2-3-3, and A2-3-4; (6) centrifuging the A2-3-4 fraction, taking the supernatant, and freeze-drying it to obtain a Radix Isatidis polypeptide, the amino acid sequence of which is shown in SEQ ID NO: 1; (7) The Radix Isatidis polypeptide shown in SEQ ID NO: 1, concentrated powder of buffalo horn, wild chrysanthemum, schizonepeta spicate, and auxiliary materials are mixed in the proportions described in claim 1 to prepare a pharmaceutical composition containing the Radix Isatidis polypeptide for anti-inflammatory, analgesic, and throat clearing.

6. Use of the anti-inflammatory, analgesic, and throat-clearing pharmaceutical composition as claimed in any one of claims 1 to 4 in the preparation of anti-inflammatory, analgesic, and throat-clearing medicines.

Images