A cockroach polypeptide site and methods of making and using same

Abstract

The present application relates to the technical field of medicine, in particular to a cockroach polypeptide fraction, a preparation method and application thereof. The present application provides a cockroach polypeptide effective fraction with a molecular weight greater than 500 Dalton (>500 Da) and less than 10,000 Dalton (<10 KDa) enriched from a fresh cockroach enzymatic product, the cockroach polypeptide effective fraction product prepared by the present application has high yield, good biological activity, and is convenient to operate and easy to operate in large-scale production. The present application further provides a cockroach polypeptide foam agent, which has a significant therapeutic effect on ulcerative colitis.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of medicine, in particular, the present application relates to a cockroach polypeptide fraction and a preparation method and application thereof. BACKGROUND

[0002] Ulcerative colitis (UC) is an immune dysfunction disease with pathological basis of mucosal epithelial cell apoptosis and inflammatory infiltration, accompanied by macrophage polarization, lymphocyte dysfunction, oxidative stress, intestinal flora imbalance and other characteristics. The clinical manifestations are mainly diarrhea, mucous and bloody stool, tenesmus, and easy to relapse. The prevalence rate in China has reached 11.6 / 100,000 people, and has a trend of increasing year by year. The first-line drugs (aminosalicylic acid) have low efficiency, long medication cycle and serious side effects, while biological agents, although effective, are expensive and have high off-target rate, which makes it difficult to be popularized in clinical practice, resulting in the quality of life of patients getting worse. Due to the characteristics of UC such as refractory and carcinogenic, it has become a research hotspot in the international digestive field, and is also a major medical and social problem that needs to be solved in China.

[0003] Although there are many drugs for treating ulcerative colitis, the clinical application effect is not satisfactory. The first-line drug aminosalicylic acid has a large first-pass effect, and has obvious side effects such as headache, nausea and gastrointestinal discomfort; glucocorticoid drugs have a large first-pass effect and low bioavailability, and long-term use can lead to metabolic disorders, decreased body resistance, gastrointestinal bacterial disorders and other side effects; and immunosuppressants have strong side effects such as bone marrow suppression and lymphoma, and are only used as auxiliary drugs for patients who cannot use aminosalicylic acid and glucocorticoid drugs; the high efficiency of biological agents brings new hope to the treatment of UC, and is mainly used for moderate and severe patients, but the high off-target rate, drug resistance and high price in clinical practice make most patients hesitant, resulting in the stagnation of the application and development of biological agents. Therefore, it is still an urgent need to strengthen the research and development of ulcerative colitis drugs to solve the current population health problems.

[0004] Periplaneta americana L. is also called Chinese medicine cockroach. Modern pharmacological studies have shown that it has the effects of antibacterial, antiviral, antitumor, immune enhancement, liver protection, and promotion of tissue repair. Its main chemical components include pheromones, protein polypeptides, amino acids, fat movement hormones, and dihydroisoflavones. In 2017, the consensus opinion on the diagnosis and treatment of ulcerative colitis in traditional Chinese and Western medicine pointed out that the ethanol extract of dried cockroach bodies, Kangfuxin liquid, has the effects of promoting blood circulation, nourishing yin, and promoting skin growth, and can be used for UC patients of various types. In clinical practice, mesalazine combined with Kangfuxin liquid, sulfasalazine combined with Kangfuxin liquid, and probiotics combined with Kangfuxin liquid can be used to treat mild to moderate UC patients through combined medication and retention enema, which can significantly improve the symptoms of abdominal pain, diarrhea, and mucous blood stool, and repair damaged intestinal mucosa.

[0005] At present, the preparation of the extract in the listed preparation of cockroaches is obtained by heating extraction with high-concentration ethanol. High-concentration ethanol not only has a killing effect on pathogenic microorganisms such as bacteria and parasites on the surface of cockroach medicinal materials during extraction, but also reduces the dissolution of macromolecular proteins and starch polysaccharides. The heating condition denatures and destroys allergen protein molecules, and at the same time, more secondary metabolites with high activity are obtained. However, during production, the safety factor of high-concentration ethanol under high-temperature heating is low, the explosion-proof performance of the production workshop is required to be high, and the yield of ethanol extract is low. The polypeptide substances with biological activity are not extracted in large quantities, resulting in waste of drug resources.

[0006] CN102743739B also discloses a polypeptide active component of cockroaches, but the component types are many and the preparation process is complex, which is not conducive to industrialization. SUMMARY

[0007] To solve the above technical problems, the present application provides a cockroach polypeptide active site for effectively treating ulcerative colitis, and a preparation method and application thereof.

[0008] The purpose of the present application is to provide a cockroach polypeptide active site with a molecular weight greater than 500 daltons (> 500 Da) and less than 10 000 daltons (< 10 KDa), which is refined from the enzymatic hydrolysate of fresh cockroach products.

[0009] The present application provides an extraction method of the cockroach polypeptide active site, which comprises the following steps: removing impurities and crushing the cockroaches, filtering after enzymatic hydrolysis, adjusting the pH, discarding the upper oil and the lower precipitate, filtering, collecting the filtrate, which is the enzymatic hydrolysate of fresh cockroach products, and loading the filtrate into a 0.5 KD dialysis bag for dialysis; after dialysis, centrifuging the solution in the bag with a 10 KD ultrafiltration centrifuge tube, collecting the lower centrifugate, and freeze-drying to obtain the cockroach polypeptide (PAP) active site.

[0010] Further, the cockroach fresh product enzymatic hydrolysate is prepared by the following method:

[0011] (1) soaking cockroaches in clean water for 2 hours;

[0012] (2) washing the residue of the sample of step (1), filtering the water on the surface of the cockroaches, adding 1-5 times the amount of pure water, and crushing into a paste for later use;

[0013] (3) mixing the paste obtained in step (2) with 3-8 times the amount of 0.1 mol / L hydrochloric acid solution, adding 0.5%-10% pepsin (m / V), stirring at 200 rpm under 37°C for 4 hours, coarsely filtering, and collecting the filtrate;

[0014] (4) adjusting the pH of the filtrate obtained in step (3) to 7.4 with 10 mM sodium hydroxide solution, adding 0.5%-10% trypsin (m / V), stirring at 200 rpm under 37°C for 2 hours, centrifuging at 8000 rpm and 4°C for 15 minutes, discarding the upper layer of oil and the lower layer of precipitate, filtering through a 0.45 μm filter head, and collecting the filtrate, thereby obtaining the cockroach fresh product enzymatic hydrolysate.

[0015] Preferably, the cockroaches in step (1) are fresh and alive or fresh bodies, and more preferably fresh and alive.

[0016] Preferably, the amount of pure water added in step (2) is 1-2 times, and more preferably 1 time.

[0017] Preferably, the mass ratio of the cockroach paste to the hydrochloric acid solution in step (3) is 1:(3-5), and more preferably 1:5.

[0018] Preferably, the proportion of pepsin in step (3) is 0.5%-3%, and more preferably 1.5%.

[0019] Preferably, the enzymatic hydrolysis time in step (3) is 2-6 hours, and more preferably 4 hours.

[0020] Preferably, the proportion of trypsin in step (4) is 0.5%-3%, and more preferably 1%.

[0021] Preferably, the enzymatic hydrolysis time in step (4) is 1-4 hours, and more preferably 2 hours.

[0022] The cockroach polypeptide effective part is prepared by the following method:

[0023] (1) placing the cockroach fresh product enzymatic hydrolysate into a 0.5 KD dialysis bag, and stirring at 300 rpm by magnetic force to dialyze;

[0024] (2) The solution in the dialysis bag in step (1) is transferred to a 10KDa ultrafiltration centrifuge tube, centrifuged at 8000rpm for 30min, the lower centrifugal liquid is collected, and freeze-dried to obtain the cockroach peptide (PAP);

[0025] Preferably, the dialysis frequency of the solution in step (1) is 1-10 times, and the single dialysis time is 1-10h; more preferably, the dialysis frequency is 4 times, and the single dialysis time is 2h.

[0026] Preferably, the ultrafiltration centrifugation temperature in step (2) is 4-25℃, the centrifugation speed is 6 000-12 000rpm, and the centrifugation time is 15-60min; more preferably, the ultrafiltration centrifugation temperature is 4-8℃, the centrifugation speed is 8 000-10 000rpm, and the centrifugation time is 30-45min.

[0027] Another object of the present application is to provide a cockroach peptide foam, which comprises the following components: 0.3-0.5 parts of the cockroach peptide active site, 0.4-0.8 parts of a foaming agent, 0.4-0.5 parts of a solubilizing agent, 1-2 parts of a thickening agent, 0.4-0.8 parts of a stabilizing agent, and 0.01-1 parts of a bacteriostatic agent.

[0028] Further, the foaming agent is selected from one or more of sodium dodecyl sulfate, sodium dodecyl sulfonate, alkyl glycoside, Tween-80, soybean phospholipid, and lecithin, preferably sodium dodecyl sulfonate or alkyl glycoside.

[0029] Further, the solubilizing agent is selected from one or more of Span-60 and poloxamer 188, preferably poloxamer 188.

[0030] Further, the thickening agent is selected from one or more of glycerol, hydroxypropyl methylcellulose (HPMC), hydroxypropyl cellulose (HPC), or carbomer 940, preferably glycerol.

[0031] Further, the stabilizing agent is selected from one or more of gum arabic, methyl cellulose, and stearic acid, preferably gum arabic or methyl cellulose.

[0032] Further, the bacteriostatic agent is selected from one or more of sodium benzoate, benzoic acid, nipagin methyl ester, and nipagin ethyl ester, preferably benzoic acid.

[0033] Further, the cockroach peptide foam comprises the following components: 0.3 parts of the cockroach peptide active site, 0.45 parts of poloxamer 188, 0.48 parts of alkyl glycoside, 1.51 parts of glycerol, 0.45 parts of gum arabic, and 0.03 parts of benzoic acid.

[0034] Further, the cockroach polypeptide foam agent comprises the following components: 0.3 parts of the cockroach polypeptide active site, 0.45 parts of poloxamer 188, 0.75 parts of sodium dodecyl sulfate, 1.51 parts of glycerol, 0.45 parts of gum arabic and 0.03 parts of benzoic acid.

[0035] Further, the cockroach polypeptide foam agent comprises the following components: 0.3 parts of the cockroach polypeptide active site, 0.45 parts of poloxamer 188, 0.75 parts of sodium dodecyl sulfate, 1.51 parts of glycerol, 0.75 parts of methyl cellulose and 0.03 parts of benzoic acid.

[0036] Further, the cockroach polypeptide foam agent is prepared as follows:

[0037] (1) Dissolve the cockroach polypeptide active site in water;

[0038] (2) Add an appropriate amount of solubilizer, mix well, and dissolve;

[0039] (3) Add an appropriate amount of foaming agent, thickening agent, stabilizer and bacteriostatic agent, and add water to the required volume;

[0040] (4) Ultrasonic for 10-60 min, homogenize, filter to get brown clear liquid, fill into container, screw on foam pump, generate foam by hand pressure emulsion pump, and get the foam.

[0041] Further, the volume expansion ratio of the cockroach polypeptide foam agent is 25-40, preferably 25-35, and most preferably 30.

[0042] Another object of the present application is to provide the use of the cockroach polypeptide active site or its composition for preparing a medicament, a daily chemical product and / or a medical device for treating ulcerative colitis.

[0043] Another object of the present application is to provide the use of the foam agent containing the cockroach polypeptide active site for preparing a medicament for treating ulcerative colitis.

[0044] The cockroach polypeptide active site of the present application is obtained by crushing and pulping fresh live insects, and then enzymatically hydrolyzing the proteins of the insects by pepsin and trypsin, and using molecular retention technology to obtain polypeptide active sites with a molecular weight greater than 0.5 KDa and less than 10 KDa, and then freeze-drying to obtain the cockroach polypeptide (PAP) product. The cockroach polypeptide active site product prepared by the present application has high yield, good biological activity, and is easy to operate and easy to produce on a large scale. The present application finds that the rectal foam enema containing the cockroach polypeptide active site has a significant therapeutic effect on ulcerative colitis.

[0045] Related definitions

[0046] The volume expansion ratio of the foam is the ratio of the volume of the expanded foam to the starting volume of the unfoamed starting material.

[0047] m / V represents mass volume ratio BRIEF DESCRIPTION OF DRAWINGS

[0048] Figure 1 Appearance of cockroach polypeptide foams prepared by adding different foaming agents, A, B: Foam No. I; C, D: Foam No. IV; E, F: Foam No. V

[0049] Figure 2 Appearance of cockroach polypeptide liposomes, A: liposomes on day 0, B: liposomes on day 3, C: liposomes on day 14

[0050] Figure 3 Colon pathological sections of mice in different administration groups, A: normal group, B: model control group, C: mesalamine group, D: PAP group, E: PAP Foam No. I group, F: PAP Foam No. II group, G: PAP Foam No. III group DETAILED DESCRIPTION

[0051] The present application will be further described in conjunction with specific examples so that those skilled in the art can better understand the present application and implement it, but the examples are not intended to limit the present application.

[0052] In the present application, the medicinal material used is fresh whole worms, and the source of the cockroaches in the present application is Periplaneta americana, and commercially available products can be used.

[0053] Example 1: Quality and activity index evaluation of cockroach polypeptides under different amounts of pepsin

[0054] Fresh cockroaches 50 g were soaked in 1 L of water for 2 h, then washed to remove residues, and the cockroaches were taken out, an equal amount of pure water was added, and the mixture was crushed. An amount of 5 times the volume of a pH 1.0 hydrochloric acid aqueous solution (0.1 mol / L) was added, and a certain proportion of pepsin (%) was added (m / V). The mixture was stirred at 37℃ under the condition of 200 rpm for 4 h. The mixture was coarsely filtered, and the residue was discarded. The filtrate was adjusted to neutral pH with 10 mM sodium hydroxide solution, and centrifuged at 8000 rpm at 4℃ for 15 min. The upper layer of oil and the lower layer of precipitate were discarded, and the filtrate was filtered through a 0.45 μm filter head. The filtrate was loaded into a 0.5 KD dialysis bag and dialyzed for 4 times, each time for 2 h. The solution in the dialysis bag was collected in a 10 KD ultrafiltration centrifuge tube, and centrifuged at 8000 rpm for 30 min. The lower layer of the centrifugate was collected, and freeze-dried to obtain cockroach polypeptide (PAP).

[0055] Table 1: Quality and activity index evaluation of cockroach polypeptides under different amounts of enzyme (pepsin)

[0056]

[0057] According to the results of Table 1, with the increase of the proportion of pepsin, the quality of the cockroaches hydrolysate gradually increased, and the in vitro activity IC 50 values gradually decreased, and when the enzyme amount exceeded 1%, there was no significant difference in in vitro changes. In order to enrich more amount of cockroach peptides, therefore, the preferred amount of pepsin enzyme is 1.5%.

[0058] Example 2 Quality and activity index evaluation of cockroach peptides under different pepsin enzymolysis time

[0059] Fresh cockroaches 50 g, soaked in 1 L water for 2 h, then washed and removed the residue, then added equal amount of pure water, crushed, added 5 times amount of pH 1.0 hydrochloric acid solution (0.1 mol / L), added 1.5% pepsin (m / V), 37°C incubation under 200 rpm stirring for a certain time, coarse filtration, discarded the residue; the filtrate was adjusted to neutral pH with 10 mM sodium hydroxide solution, centrifuged at 8000 rpm at 4°C for 15 min, discarded the upper layer of oil and the lower layer of precipitate, filtered through a 0.45 μm filter head, the filtrate was loaded into a 0.5 KD dialysis bag and dialyzed for 4 times, each time for 2 h; the solution in the dialysis bag was centrifuged at 8000 rpm for 30 min in a 10 KD ultrafiltration centrifuge tube, the lower layer of centrifugate was collected, freeze-dried, and cockroach peptides (PAP) were obtained.

[0060] Table 2 Quality and activity index evaluation of cockroach peptides under different pepsin enzymolysis time

[0061]

[0062] According to the results of Table 2, with the extension of the enzymolysis time, the quality of the cockroach hydrolysate first increased and then decreased, and the in vitro activity IC 50 values first decreased and then increased, and when the enzymolysis time exceeded 4 h, the content of cockroach peptides decreased and the in vitro activity decreased. In order to enrich more amount of active cockroach peptides, the preferred pepsin enzymolysis time is 4 h.

[0063] Example 3 Yield and activity index evaluation of cockroach hydrolysate under different amounts of trypsin

[0064] Fresh cockroaches 50g, soaked in 1L water for 2h, then washed and removed the residue, added equal amount of pure water, crushed, added 5 times amount of pH 1.0 hydrochloric acid solution (0.1mol / L), added 1.5% pepsin (m / V), 37℃ incubation under 200rpm stirring for 4h, coarse filtration, discarded the residue; the filtrate was adjusted to pH 7.4 with 10mM sodium hydroxide solution, added a certain proportion of trypsin (% m / V), 37℃ incubation under 200rpm stirring for 2h, 8000rpm 4℃ centrifugation for 15min, discarded the upper oil and the lower precipitate, 0.45μm filter head filtration, the filtrate was loaded into 0.5KD dialysis bag for dialysis for 4 times, 2h each time; the solution in the dialysis bag was centrifuged in 10KD ultrafiltration centrifuge tube under 8000rpm for 30min, the lower centrifugal liquid was collected, freeze-dried, and cockroach polypeptide (PAP) was obtained.

[0065] Table 3 yield and activity index evaluation of cockroach enzymatic hydrolysate under different enzyme amount (trypsin)

[0066]

[0067] According to the results in Table 3, with the increase of trypsin proportion, the quality of cockroach polypeptide increased first and then decreased, the in vitro activity IC 50 value decreased first and then increased, the quality of cockroach polypeptide gradually increased, and the in vitro activity IC 50 value gradually decreased, when the enzyme amount exceeded 1%, the quality and activity of polypeptide decreased. Therefore, the preferred trypsin enzyme amount was 1.0%.

[0068] Example 4 quality and activity index evaluation of cockroach polypeptide under different enzymolysis time of trypsin

[0069] Fresh cockroaches 50g, soaked in 1L water for 2h, then washed and removed the residue, added equal amount of pure water, crushed, added 5 times amount of pH 1.0 hydrochloric acid solution (0.1mol / L), added 1.5% pepsin (m / V), 37℃ incubation under 200rpm stirring for 4h, coarse filtration, discarded the residue; the filtrate was adjusted to pH 7.4 with 10mM sodium hydroxide solution, added 1.0% trypsin (% m / V), 37℃ incubation under 200rpm stirring for a certain time, 8000rpm 4℃ centrifugation for 15min, discarded the upper oil and the lower precipitate, 0.45μm filter head filtration, the filtrate was loaded into 0.5KD dialysis bag for dialysis for 4 times, 2h each time; the solution in the dialysis bag was centrifuged in 10KD ultrafiltration centrifuge tube under 8000rpm for 30min, the lower centrifugal liquid was collected, freeze-dried, and cockroach polypeptide (PAP) was obtained.

[0070] Table 4 quality and activity index evaluation of cockroach polypeptide under different enzymolysis (trypsin) time

[0071]

[0072] According to the results of Table 4, with the extension of enzymolysis time, the quality of cockroach polypeptide increased first and then decreased, and the in vitro activity IC 50 value decreased first and then increased. When the enzymolysis time was 2 h, the quality of cockroach polypeptide was the highest, and the in vitro activity IC 50 value was the lowest. In order to enrich more amount of cockroach polypeptide, therefore, the enzymolysis time of trypsin is preferably 2 h.

[0073] Example 5 Quality and activity index evaluation of cockroach extract obtained by different preparation processes

[0074] Fresh cockroaches 50 g were dried at 50°C, crushed, soaked in 3 times petroleum ether at room temperature for 24 h, repeated soaking for 3 times, and the cockroach bodies were taken out and dried at 60°C for standby. 5 times amount of distilled water and 1.0% trypsin (%, m / V) were added to the cockroach powder, and stirred at 200 rpm under the condition of 37°C for 2 h, then the temperature was increased to 80°C to kill the trypsin, and after cooling, filtration was performed, and the filtrate was dried at 40°C to obtain the cockroach extract (PAP-11).

[0075] Fresh cockroaches 50 g were dried at 50°C, crushed, soaked in 3 times petroleum ether at room temperature for 24 h, repeated soaking for 3 times, and the cockroach bodies were taken out and dried at 60°C for standby. 5 times amount of distilled water and 1.0% trypsin (%, m / V) were added to the cockroach powder, and stirred at 200 rpm under the condition of 37°C for 2 h, then the temperature was increased to 80°C to kill the trypsin, and after cooling, filtration was performed, and the filtrate was dried at 40°C to obtain the cockroach extract (PAP-11).

[0076] Table 5 Quality and activity index evaluation of cockroach extract obtained by different preparation processes

[0077]

[0078] According to the results of Table 5, the quality and in vitro activity of cockroach polypeptide obtained by different preparation processes were found. The content of cockroach polypeptide obtained by trypsin enzymolysis process (PAP11) was significantly higher than that obtained by water extraction and alcohol precipitation process (PAP12), but the IC 50 value of the in vitro anti-inflammatory, antioxidant and repair-promoting ability of the extract was higher, and the activity was weaker than that of the water extraction and alcohol precipitation process, and also weaker than that of the active polypeptide fraction in other examples.

[0079] Example 6 Preparation and evaluation of blank foam

[0080] Dissolve 0.45 g of poloxamer 188 in water, then add 1.20 mL of glycerin, 0.48 g of alkyl glycoside 0810, 0.45 g of gum arabic, 0.3 g of benzoic acid, and make up to 15 mL with water, homogenize for 30 min with ultrasonic, filter to get a brown clear liquid, fill into a container, screw on a foam pump, generate foam by hand-pressing the emulsion pump, and the blank foam is obtained.

[0081] Evaluation method of blank foam:

[0082] The existing external use foam belongs to pharmaceutical aerosol, and should meet the quality standards of aerosol itself. However, due to the unique physical properties of foam, in the process of prescription and technology development, appropriate performance evaluation indexes should be used according to the particularity of foam, so as to better meet the quality standards of external use foam. Foamability and foam stability are the two most basic indexes in the evaluation of foam. Among them, foamability is usually characterized by parameters such as foaming amount, foaming ratio and foaming time, which reflect the size of foam amount and whether the foam is easy to form. The stability of foam refers to the persistence of foam generation, that is, the difficulty of defoaming, which is usually characterized by half-life.

[0083] Comprehensive consideration, the micro-morphology, foam density, expansion rate, defoaming rate and comprehensive index of foam are weighted and scored according to the weight coefficient of 10, 10, 20, 20 and 40.

[0084] Table 6 Evaluation method of foam

[0085]

[0086] Note: The appearance score includes the uniformity of the original solution, the toughness of the foam, the fineness of the foam and the distribution of the foam. The uniformity of the original solution: the original solution has insoluble matter and is opaque, which is poor, the original solution has no insoluble matter but is not clear, which is general, and the original solution has no insoluble matter and is uniform and clear, which is good; the toughness of the foam: the foam is dry and easy to break by hand rubbing, which is poor, the foam is slightly moist, and the foam diameter becomes larger and does not break after long rubbing, which is general, and the foam is moist, and the shape of the foam does not change much after long rubbing, which is good; the fineness of the foam: the single foam is obvious and visible with large diameter, which is poor, the single foam is small in diameter and can be distinguished, which is general, and the single foam is not easy to distinguish, and the diameter cannot be measured, which is good; the distribution of the foam: the foam liquid film is thin, and the liquid is almost concentrated in the Plateau channel, which is poor, the gas bubble distribution is appropriate, the foam liquid film is thick, which is general, and the gas bubble distribution is narrow and uniform, and the liquid in the foam is evenly distributed in the liquid film and the Plateau channel, which is good.

[0087] Foam density = m 泡 / v 泡 Foam expansion rate = (v 泡 -v 液 ) / v 液

[0088] Foam defoaming rate = (hi - h2) / hi * 100% Foam comprehensive index = v 泡 *t 半衰期

[0089] Wherein: m 泡 is the mass of the foam emitted, v 泡 is the corresponding foam volume;

[0090] v 液 is the volume of liquid required to form the corresponding volume of foam;

[0091] hi is the initial height of the foam, h2 is the height of the foam after 10 min;

[0092] t 半衰期 is the time required for the foam volume to decay by half.

[0093] Example 7 Preparation of PAP Foam I

[0094] Take cockroach polypeptide (PAP) 0.3 g dissolved in water; add 0.45 g of poloxamer 188, mix, dissolve, then add 1.20 mL of glycerol, 0.48 g of alkyl glycoside 0810, 0.45 g of gum arabic, 0.03 g of benzoic acid, make up to 15 mL with water, homogenize by ultrasonic for 30 min, filter to obtain a brown clear liquid, fill into a container, screw on a foam pump, generate foam by hand pressing the emulsion pump, and PAP Foam I is obtained.

[0095] Example 8 Preparation of PAP Foam II

[0096] Take cockroach polypeptide (PAP) 0.3 g dissolved in water; add 0.45 g of poloxamer 188, mix, dissolve, then add 1.20 mL of glycerol, 0.48 g of alkyl glycoside 0810, 0.45 g of gum arabic, 0.03 g of benzoic acid, make up to 15 mL with water, homogenize by ultrasonic for 30 min, filter to obtain a brown clear liquid, fill into a container, screw on a foam pump, generate foam by hand pressing the emulsion pump, and PAP Foam I is obtained.

[0097] Example 9 Preparation of PAP Foam III

[0098] Take cockroach polypeptide (PAP) 0.3 g dissolved in water; add 0.45 g of poloxamer 188, mix, dissolve, then add 1.20 mL of glycerol, 0.48 g of alkyl glycoside 0810, 0.45 g of gum arabic, 0.03 g of benzoic acid, make up to 15 mL with water, homogenize by ultrasonic for 30 min, filter to obtain a brown clear liquid, fill into a container, screw on a foam pump, generate foam by hand pressing the emulsion pump, and PAP Foam I is obtained.

[0099] The physical properties of PAP foam agents I to III are compared according to the evaluation method for foam agents, as shown in Table 7.

[0100] Table 7. Comprehensive Evaluation of PAP Foaming Agents I-III

[0101]

[0102] Comparative Example 1

[0103] Dissolve 0.3g of cockroach polypeptide (PAP) in water; add 0.45g of poloxamer 188, mix well to dissolve, then add 1.20mL of glycerol, 0.15g of Tween-80, 0.45g of gum arabic, and 0.03g of benzoic acid. Add water to make up to 15mL, sonicate for 30min to homogenize, filter to obtain a clear brown liquid, pour into a container, screw on the cap and attach a foam pump, and generate foam by hand-pressing the emulsion pump to obtain PAP foam agent No. IV.

[0104] Comparative Example 2

[0105] Dissolve 0.3g of cockroach polypeptide (PAP) in water; add 0.45g of poloxamer 188, mix well to dissolve, then add 1.20mL of glycerol, 0.15g of soybean lecithin, 0.45g of gum arabic, and 0.03g of benzoic acid. Add water to make up to 15mL, sonicate for 30min to homogenize, filter to obtain a clear brown liquid, pour into a container, screw on the cap and attach a foam pump, and generate foam by hand-pressing the emulsion pump to obtain PAP foam agent No. V.

[0106] Compare the differences in physical properties of PAP foam agents I, IV, and V based on the evaluation methods for foam agents:

[0107] Table 8. Comprehensive Evaluation of PAP Foaming Agents I, IV, and V

[0108]

[0109] Replace the foaming agent alkyl glycoside (APG0810) in the formula with an equal amount of Tween-80 and soybean lecithin, keeping other ingredients unchanged, to create a foaming agent. For example... Figure 1 As shown, based on appearance, the solution of PAP foaming agent I is clear, the solution of foaming agent IV is viscous and has poor clarity, and the solution of foaming agent V is cloudy. PAP foaming agent I produces rich, fine, and uniform foam, while foaming agent IV produces foam with obvious pores and stratification. Foaming agent V's foam dissipates instantly and hardly forms any foam. Based on the weighted score of the foaming agents, the result is: PAP foaming agent I > PAP foaming agent IV > PAP foaming agent V. Therefore, not all surfactants can form strong and stable foams with other excipients.

[0110] Comparative Example 3

[0111] Precisely weigh 12 mg of soybean lecithin and 2.4 mg of cholesterol, respectively, and place them in a 100 mL tomato bottle. Add 5 mL of dichloromethane to dissolve them thoroughly. Rotate the bottle on a rotary evaporator at 200 rpm to remove the dichloromethane, and a uniform lipid film is formed on the bottle wall. Weigh 3.0 mg of PAP and prepare a 1.0 mg / mL solution with PBS (pH = 7.4). Pour the solution into the above-mentioned tomato bottle and shake it on a constant temperature shaker at 30°C and 200 rpm for 0.5 h to obtain the Lipo-PAP primary product. The Lipo-PAP primary product is extruded twice through a 0.20 μm microporous filter membrane liposome extruder to obtain a milky Lipo-PAP suspension, which is the PAP liposome.

[0112] Place PAP foam agent No. I and PAP liposome in a room temperature environment for 14 days, and observe the differences in appearance and stability of the two. After 14 days in a room temperature environment, the PAP foam agent No. I solution is clear and transparent, and no precipitate is produced. However, the liposome has a slight white precipitate from the third day, and a large amount of white flocculent precipitate is produced on the 14th day. Figure 2 )。

[0113] Test Example 1 Treatment effect of cockroach polypeptide foam on ulcerative colitis mice by in situ enema

[0114] 1. Experimental materials

[0115] 1.1 Experimental animals

[0116] Forty-eight healthy SPF C57BL / 6 mice, male, 6-8 weeks old, body weight 18-22 g, purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd., license number: SCXK (Jing) 2021-0006; raised in the IVC system of the Animal Experimental Center of Nanjing University of Chinese Medicine, temperature 20-25°C, humidity 40-70% RH, 12 h light and dark alternation, experimental unit use license number: SYXK (Su) 2018-0049.

[0117] 1.2 Test drugs

[0118] The blank foam agent, PAP, PAP foam agent No. I, PAP foam agent No. II, and PAP foam agent No. III of the present application;

[0119] Mesalazine (batch number: 1902002, Tianjin Lisheng Pharmaceutical Co., Ltd., specification: 0.25 g).

[0120] 1.3 Experimental reagents

[0121] Dextran sulfate sodium (batch number: 11032-222, Shenzhen Lijing Company);

[0122] Anhydrous ethanol, AR (Batch number: 2017, Tianjin Fuchen Chemical Reagent Factory);

[0123] Isoflurane, (Batch number: 20190701, Shandong Keyuan Pharmaceutical Co., Ltd.);

[0124] Chloral hydrate, AR (Batch number: 20170203, National Pharmaceutical Group Chemical Reagent Co., Ltd.);

[0125] Formaldehyde solution, AR (Batch number: 20170523, Tianjin Fushen Chemical Reagent Technology Co., Ltd.);

[0126] Disodium hydrogen phosphate, AR (Batch number: 20171008, Tianjin Fushen Chemical Reagent Technology Co., Ltd.);

[0127] Sodium dihydrogen phosphate, AR (Batch number: 20160318, Tianjin Fushen Chemical Reagent Technology Co., Ltd.);

[0128] Normal saline, NS (Batch number: A15031602, Guizhou Tiandi Pharmaceutical Co., Ltd.);

[0129] Hidden blood kit, (Batch number: 20190323, Nanjing Jianshen Biological Engineering Institute);

[0130] Mouse interleukin 6 (IL-6) kit, batch number: 20220207-20188A; Mouse interleukin 17A (IL-17A) kit, batch number: 20220207-20171A; Mouse myeloperoxidase (MPO) kit, batch number: 20220207-28122A; Mouse epidermal growth factor (EGF) kit, batch number: 20220207-20203A; All purchased from Shanghai Zoben Biological Technology Co., Ltd.

[0131] 1.4 Main instruments

[0132] High speed centrifuge (Anhui Zhongke Zhongjia Science and Technology Instrument Co., Ltd.), Model: HC-30118R;

[0133] Electronic balance (Mettler-Toledo Instrument Co., Ltd.), Model: AL204-IC;

[0134] Precise electronic analytical balance (Mettler-Toledo Instrument Co., Ltd.), Model: METTLER AE240;

[0135] Vortex instrument (Haimen Qishenbel Instrument Manufacturing Co., Ltd.), Model: VORT FX-5;

[0136] Freeze dryer (Cold-Sim Company, USA), Model: FD8-4a;

[0137] Common optical microscope (Chongqing Optical Instrument Factory), model: OLYMPUS CX31;

[0138] Pathological tissue float-drying instrument (Changzhou Zhongwei Electronic Instrument Co., Ltd.), model: PHY-III;

[0139] Paraffin section machine (Leica Company), model: RM2245;

[0140] Freezing table (Changzhou Zhongwei Electronic Instrument Co., Ltd.), model: BMJ-C;

[0141] Biological tissue embedding machine (Xiaogan Hongye Medical Instrument Co., Ltd.), model: BM-VIII;

[0142] Biological tissue float-drying machine (Hubei Xiaogan Yaguang Medical Electronic Technology Co., Ltd.), model: YT-7FB;

[0143] Automatic dehydration machine (produced by Germany Leica Company), model: Leica ASP-300S;

[0144] Medical image analysis system (Chengdu Tailian Technology Co., Ltd.), model: BI-2000;

[0145] Macro camera, model: DIGITAI CAMERA D7100, macro lens, model: AF-S Micro 60 / 2.8G ED, all purchased from Nikon.

[0146] 2. Experimental method

[0147] 2.1 Establishment of experimental animal model

[0148] After C57BL / 6 mice were adaptively fed for 7 days, except that the normal control group was given pure water, the rest of the mice were given 3% (w / v) dextran sulfate sodium salt (DSS) aqueous solution for free drinking for 7 days, the solution was changed every 2 days, and after the modeling was completed, the mice were fed with pure water for 7 days, a total of 14 days.

[0149] 2.2 Experimental animal grouping and administration

[0150] Randomly selected 6 mice as normal control group, the remaining mice were modeled according to the above method, on the 7th day of modeling, the model mice were scored by disease activity index (Disease activity index, DAI) (0-3 points for mild inflammation, 4-6 points for mild inflammation, 7-9 points for moderate inflammation, 10-12 points for severe inflammation), after excluding the mice with mild inflammation, the remaining mice were divided into model control group, mesalamine group (200mg / kg), PAP group, PAP foam agent I group, PAP foam agent II group, PAP foam agent III group according to the method of DAI stratification and random sampling, 6 in each group. The next day of grouping, each administration group was given 0.1 mL / 20g·d enema with corresponding drugs, and the normal control group and the model control group were given 0.1 mL / 20g·d blank foam agent, for 7 consecutive days.

[0151] 2.3 Index observation and detection method

[0152] 2.3.1 Disease activity index

[0153] The body weight of mice was weighed every day during the experiment, the stool characteristics and fecal occult blood of mice were observed, and the DAI score of each mouse was calculated to evaluate the disease activity.

[0154] 2.3.2 Colon mucosal damage index

[0155] After the last administration, the mice were fasted for 24 hours without water, and then executed by cervical dislocation, and the entire colon from about 1cm from the anus to the end of the cecum was cut off, laid on a white porcelain plate, measured the length of the colon with a steel ruler, cut off along the mesentery, washed the lumen contents with ice physiological saline, and scored the colon mucosal damage index (Colon mucocal damage index, CMDI) according to the literature standard.

[0156] 2.3.3 Histological score

[0157] Half of the colon tissue was cut longitudinally, fixed with 10% (W / V) neutral formalin, dehydrated with paraffin, and embedded with paraffin to make 5μm pathological sections, stained with hematoxylin-eosin (HE), sealed with neutral gum, observed the pathological changes under a microscope, and scored the colon histological score (Histoligical score, HS) according to the literature standard.

[0158] 2.3.4 Cytokine determination of colon tissue

[0159] The other half of the colon tissue was longitudinally cut, and a 10% (W / V) tissue homogenate was prepared with pre-cooled normal saline. The mixture was centrifuged at 3000 r / min for 10 min at 4 ℃, and the supernatant was collected. The contents of TNF-α, IL-6, MPO, and EGF in the colon tissue were detected according to the relevant kit operation manual.

[0160] 3. Results

[0161] 3.1 Effect on the general condition of ulcerative colitis (UC) mice

[0162] The normal control group of mice had normal diet and water, smooth and clean hair, good mental state, feces with black color, hard texture and ellipsoidal shape, and urine with light yellow color and normal color. The mental state of the model control group of mice improved over time, and the food intake increased slightly, but the tail still attached to the loose stool, or the stool with blood. The mental state of the mesalazine group of mice improved significantly, and the activity increased, and there was no yellow sticky stool. The feces were basically shaped and turned black. The mental state of the PAP group of mice improved, the phenomenon of huddling and curling decreased, the anus was less dirty, the feces were basically shaped and the color turned black, and the body weight gradually increased. The mental state of the PAP foam I, II and III groups of mice was good, the activity increased, the anus had no sticky stool, the feces were shaped and the color turned black, and the body weight gradually increased.

[0163] 3.2 Effect on the DAI score of UC mice

[0164] Mauchly's sphericity test showed that the DAI score data of each group of mice from day 0 to day 14 did not conform to the sphericity assumption (P<0.05), so the results were analyzed after correcting the degrees of freedom by Greenhousc-Geisser method. The results of repeated measurement analysis of variance showed that: (1) interaction (time*group): the interaction of DAI score of mice was statistically significant (P<0.01), which indicated that the effect of time factor on DAI score of mice in different groups was different. (2) Time effect: the time effect of DAI score of mice was statistically significant (P<0.01), which indicated that the DAI score of mice at different time points was different. (3) Group effect: the group effect of DAI score of mice was statistically significant (P<0.01), which indicated that the DAI score of mice in each group was different. In order to evaluate the group difference at the same time point, according to the results of single factor analysis of variance, on the 0th day of the experiment, the DAI scores of each group were low and had no significant difference (P>0.05), that is, all mice were normal and at the same level. On the 7th day of the experiment, the last day of modeling, the DAI score of the normal control group was low, the DAI score of all the modeling mice was significantly higher than that of the normal group (P<0.01), and there was no significant difference between each modeling group, which indicated that the UC model was successfully replicated. On the 14th day of the experiment, the drug treatment ended, compared with the normal control group, the DAI scores of the model control group and each drug treatment group were significantly increased (P<0.01), compared with the model control group, all treatment groups could effectively reduce the DAI score (P<0.01), among which except for group I compared with mesalazine group and PAP group respectively (P<0.05), the rest of the treatment groups had no obvious difference, and the DAI scores were significantly reduced.

[0165] Table 9 Effect of PAP on DAI score of UC mice n=6

[0166]

[0167] Note: compared with the normal group, * P<0.05, ** P<0.01; compared with the model group, △ P<0.05, △△ P<0.01; compared with the mesalazine group, ▲ P<0.05, ▲ P<0.01; compared with the PAP group, ▼ P<0.05, ▼▼ P<0.01.

[0168] 3.3 Effect on colon-related indicators of UC mice

[0169] The colon mucosa of normal control mice showed no congestion, edema, erosion and ulcer. The intestinal wall of model control mice showed congestion, edema and thickening, and the intestinal mucosa showed necrosis and ulcer formation, with the most severe lesions at the anal end of the colon. The colon mucosa of mice in each drug treatment group showed some improvement. The mesalamine group showed congestion and edema, and the intestinal wall showed small superficial ulceration. The PAP group showed mild congestion and intestinal wall thickening. The foam agent groups I, II and III showed varying degrees of relief of congestion, edema and ulceration. Compared with the normal control group, the colon length of the model control group was significantly shorter (P<0.01), and the CMDI score was significantly higher (P<0.01). Compared with the model control group, the colon length of all treatment groups was significantly longer (P<0.05 or P<0.01), and the CMDI score was significantly lower (P<0.05 or P<0.01). Compared with the mesalamine group, the colon length and CMDI score of the foam agent group I showed no significant difference (P>0.05), and the colon length and CMDI score of the foam agent groups II and III showed no significant difference compared with the PAP group (P>0.05).

[0170] Table 10 Effect of PAP on colon-related indicators in UC mice n = 6

[0171]

[0172] Note: Compared with the normal group, * P<0.05, ** P<0.01; compared with the model group, △ P<0.05, △△ P<0.01; compared with the mesalamine group, ▲ P<0.05, ▲ P<0.01; compared with the PAP group, ▼ P<0.05, ▼▼ P<0.01.

[0173] As Figure 3As shown, under the microscope, the mucosa and its underlying structure of the normal control group were complete and clear, the glands were arranged in order, and obvious crypts and goblet cells were observed, without hyperemia, edema, and inflammatory cell infiltration. In the model control group, the colonic mucosa was shed and damaged, and a large number of inflammatory cells infiltrated into the mucosa muscular layer and muscular layer. In the mesalamine group, the PAP group, and the foam agent I, II, and III groups, the colonic goblet cells and crypts were improved to different degrees, and the inflammatory cell infiltration was significantly reduced. Compared with the normal control group, the epithelial cell score, inflammatory cell infiltration score, and HS total score of the model control group were significantly increased (P<0.01). Compared with the model control group, the epithelial cell score, inflammatory cell infiltration score, and HS total score of each treatment group were reduced to different degrees (P<0.01). Among them, the epithelial cell integrity and total score evaluation of the foam agent I group were significantly better than those of the mesalamine group and the PAP group (P<0.05).

[0174] Table 11 Effect of PAP on the HS score of the colon of UC mice n = 6

[0175]

[0176]

[0177] Note: Compared with the normal group, * P<0.05, ** P<0.01; compared with the model group, △ P<0.05, △△ P<0.01; compared with the mesalamine group, ▲ P<0.05, ▲▲ P<0.01; compared with the PAP group, ▼ P<0.05, ▼▼ P<0.01.

[0178] 3.4 Effect on cytokines in the colon tissue of UC mice

[0179] Compared with the normal group, the expression levels of inflammatory factors IL-17A, IL-6, and oxidative stress-related activating enzyme MPO were significantly increased in the colonic tissue of mice in the model group (P < 0.01), while the expression level of tissue repair-related factor EGF was significantly decreased (P < 0.01). Compared with the model group, the expression levels of inflammatory factors IL-17A, IL-6, and MPO were significantly decreased in the colonic tissue of mice in each treatment group (P < 0.01), while the expression level of EGF was significantly increased (P < 0.01). Among them, the foam preparation group I showed the most significant effect in inhibiting the expression of inflammatory factors IL-17A and IL-6 and reducing MPO content (P < 0.01), and the effect was more significant compared with the mesalazine group and the PAP group (P < 0.05 or P < 0.01). This indicates that the PAP fraction of cockroach polypeptide has a therapeutic effect on UC mice, and the effect is more significant after it is prepared into a foam preparation.

[0180] Table 12 Effects of PAP drug groups on cytokines in colon tissue of UC mice ( n=6)

[0181]

[0182] Note: Compared with the normal group, * P < 0.05 ** P < 0.01; compared with the model group, △ P < 0.05 △△ P < 0.01; compared with the mesalazine group, ▲ P < 0.05 ▲ P < 0.01; compared with the PAP group, ▼ P < 0.05 ▼▼ P < 0.01.

[0183] The results of this study suggest that the PAP fraction of cockroaches, administered via enema, can improve the disease activity index in mice, promote colonic mucosal repair, and reduce the secretion of inflammatory factors, thus exerting its therapeutic effect on ulcerative colitis (UC).

Claims

1. Use of a cockroach polypeptide active site with a molecular weight greater than 500 daltons and less than 10,000 daltons or a composition thereof for the preparation of a medicament for treating ulcerative colitis, The cockroach polypeptide active site is prepared as follows: 1) Fresh cockroaches are soaked in clean water for 2 h; 2) The sample of step 1) is washed to remove residues, the surface moisture of the cockroaches is filtered dry, an amount of pure water equal to the amount of water used in step 1) is added, and the mixture is crushed into a paste for use; 3) The paste obtained in step 2) is mixed with 5 times the amount of 0.1 mol / L hydrochloric acid solution, then 0.5% to 1.5% (m / V) pepsin is added, and the mixture is stirred at 200 rpm under 37°C for 2 to 6 h, then the mixture is coarsely filtered, and the filtrate is collected; 4) The filtrate obtained in step 3) is adjusted to pH 7.4 with 10 mM sodium hydroxide solution, then 0.5% to 1.5% (m / V) trypsin is added, and the mixture is stirred at 200 rpm under 37°C for 1 to 4 h, then the mixture is centrifuged at 8000 rpm and 4°C for 15 min, the upper layer of oil and the lower layer of precipitate are discarded, the mixture is filtered through a 0.45 μm filter head, and the filtrate is collected, thereby obtaining a cockroach fresh product enzymatic product; The cockroach polypeptide active site is extracted as follows: S1: The cockroach fresh product enzymatic product is placed in a 0.5 KD dialysis bag, and dialysis is performed 4 times, each time for 2 h; S2: The solution in the dialysis bag in step S1 is transferred to a 10 KDa ultrafiltration centrifuge tube, and the mixture is centrifuged at 8000 rpm and 4°C for 30 min, then the lower layer of centrifugate is collected, and the mixture is freeze-dried, thereby obtaining a cockroach polypeptide active site.

2. Use of a cockroach polypeptide foam prepared from a cockroach polypeptide active site with a molecular weight greater than 500 daltons and less than 10,000 daltons for the preparation of a medicament for treating ulcerative colitis; wherein, The cockroach polypeptide active site is prepared as follows: 1) Fresh cockroaches are soaked in clean water for 2 h; 2) The sample of step 1) is washed to remove residues, the surface moisture of the cockroaches is filtered dry, an amount of pure water equal to the amount of water used in step 1) is added, and the mixture is crushed into a paste for use; 3) The paste obtained in step 2) is mixed with 5 times the amount of 0.1 mol / L hydrochloric acid solution, then 0.5% to 1.5% (m / V) pepsin is added, and the mixture is stirred at 200 rpm under 37°C for 2 to 6 h, then the mixture is coarsely filtered, and the filtrate is collected; 4) The filtrate obtained in step 3) is adjusted to pH 7.4 with 10 mM sodium hydroxide solution, then 0.5% to 1.5% (m / V) trypsin is added, and the mixture is stirred at 200 rpm under 37°C for 1 to 4 h, then the mixture is centrifuged at 8000 rpm and 4°C for 15 min, the upper layer of oil and the lower layer of precipitate are discarded, the mixture is filtered through a 0.45 μm filter head, and the filtrate is collected, thereby obtaining a cockroach fresh product enzymatic product; The cockroach polypeptide active site is extracted as follows: S1: The cockroach fresh product enzymatic product is placed in a 0.5 KD dialysis bag, and dialysis is performed 4 times, each time for 2 h; S2: The solution in the dialysis bag in step S1 is transferred to a 10 KDa ultrafiltration centrifuge tube, and the mixture is centrifuged at 8000 rpm and 4°C for 30 min, then the lower layer of centrifugate is collected, and the mixture is freeze-dried, thereby obtaining a cockroach polypeptide active site. S2: the solution in the dialysis bag in step S1 is transferred to a 10 KDa ultrafiltration centrifuge tube, centrifuged at 8000 rpm at 4℃ for 30 min, the lower centrifugal liquid is collected, and freeze-drying is performed, to obtain the cockroach polypeptide effective part; The cockroach polypeptide foam agent comprises the following components in parts by weight: 0.3 parts of the cockroach polypeptide effective part, 0.45 parts of poloxamer 188, 0.48 parts of alkyl polyglycoside, 1.51 parts of glycerol, 0.45 parts of gum arabic and 0.03 parts of benzoic acid.

3. Use of a cockroach polypeptide foam agent prepared from a cockroach polypeptide effective part with a molecular weight greater than 500 daltons and less than 10,000 daltons for preparing a drug for treating ulcerative colitis; wherein, The cockroach polypeptide effective part is prepared by the following method: 1) Fresh cockroaches are soaked in clean water for 2 h; 2) The sample of step 1) is washed to remove residues, the surface moisture of the cockroaches is filtered and dried, 1 times the amount of pure water is added, and the mixture is crushed into a paste for standby; 3) The paste obtained in step 2) is mixed with 5 times the amount of 0.1 mol / L hydrochloric acid solution, 0.5%~1.5% pepsin is added, and the mixture is stirred at 200 rpm at 37℃ for 2~6 h, then the mixture is coarsely filtered, and the filtrate is collected; 4) The filtrate obtained in step 3) is adjusted to pH 7.4 with 10 mM sodium hydroxide solution, 0.5%~1.5% trypsin is added, and the mixture is stirred at 200 rpm at 37℃ for 1~4 h, then the mixture is centrifuged at 8000 rpm at 4℃ for 15 min, the upper oil and the lower precipitate are discarded, the mixture is filtered through a 0.45 μm filter head, and the filtrate is collected, to obtain a cockroach fresh product enzymatic product; The cockroach polypeptide effective part is extracted by the following method: S1: The cockroach fresh product enzymatic product is loaded into a 0.5 KD dialysis bag, and dialysis is performed 4 times, each time for 2 h; S2: The solution in the dialysis bag in step S1 is transferred to a 10 KDa ultrafiltration centrifuge tube, centrifuged at 8000 rpm at 4℃ for 30 min, the lower centrifugal liquid is collected, and freeze-drying is performed, to obtain the cockroach polypeptide effective part; The cockroach polypeptide foam agent comprises the following components: 0.3 parts of the cockroach polypeptide effective part, 0.45 parts of poloxamer 188, 0.75 parts of sodium dodecyl sulfate, 1.51 parts of glycerol, 0.45 parts of gum arabic and 0.03 parts of benzoic acid.

4. Use of a cockroach polypeptide foam agent prepared from a cockroach polypeptide effective part with a molecular weight greater than 500 daltons and less than 10,000 daltons for preparing a drug for treating ulcerative colitis; wherein, The cockroach polypeptide effective part is prepared by the following method: 1) Fresh cockroaches are soaked in clean water for 2 h; 2) The sample of step 1) is washed to remove residues, the surface moisture of the cockroaches is filtered and dried, 1 times the amount of pure water is added, and the mixture is crushed into a paste for standby; 3) The paste obtained in step 2) is mixed with 5 times the amount of 0.1 mol / L hydrochloric acid solution, 0.5%~1.5% pepsin is added, and the mixture is stirred at 200 rpm at 37℃ for 2~6 h, then the mixture is coarsely filtered, and the filtrate is collected; 4) the filtrate obtained in step 3) is adjusted to pH 7.4 with 10 mM sodium hydroxide solution, 0.5%~1.5% trypsin by mass is added, and the mixture is incubated at 37℃ with stirring at 200 rpm for 1~4 h, then centrifuged at 8000 rpm at 4℃ for 15 min, the upper layer of oil and the lower layer of precipitate are discarded, and the filtrate is collected by filtration through a 0.45 μm filter, thereby obtaining the cockroach fresh product enzymatic hydrolysate; The cockroach polypeptide effective part extraction method is as follows: S1: the cockroach fresh product enzymatic hydrolysate is loaded into a 0.5 KD dialysis bag, and dialysis is performed 4 times, each time for 2 h; S2: the solution in the dialysis bag in step S1 is transferred to a 10 KDa ultrafiltration centrifuge tube, centrifuged at 8000 rpm at 4℃ for 30 min, and the lower layer of centrifugate is collected and freeze-dried, thereby obtaining the cockroach polypeptide effective part; The cockroach polypeptide foam agent comprises the following components: 0.3 parts of the cockroach polypeptide effective part, 0.45 parts of poloxamer 188, 0.75 parts of sodium dodecyl sulfate, 1.51 parts of glycerol, 0.75 parts of methyl cellulose, and 0.03 parts of benzoic acid.

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