CN114099640A - Antibacterial peptide composition for inhibiting and killing helicobacter pylori and preparation method and application thereof - Google Patents

Antibacterial peptide composition for inhibiting and killing helicobacter pylori and preparation method and application thereof Download PDF

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CN114099640A
CN114099640A CN202111527966.7A CN202111527966A CN114099640A CN 114099640 A CN114099640 A CN 114099640A CN 202111527966 A CN202111527966 A CN 202111527966A CN 114099640 A CN114099640 A CN 114099640A
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nisin
antibacterial peptide
peg
composition
helicobacter pylori
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王天琪
张庆友
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Likang Rongjian Beijing Biomedical Technology Co ltd
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Likang Rongjian Beijing Biomedical Technology Co ltd
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/164—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/17—Amino acids, peptides or proteins
    • A23L33/18—Peptides; Protein hydrolysates
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/56—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule
    • A61K47/59—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes
    • A61K47/60—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes the organic macromolecular compound being a polyoxyalkylene oligomer, polymer or dendrimer, e.g. PEG, PPG, PEO or polyglycerol
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/04—Antibacterial agents
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs

Abstract

The invention provides a composition for inhibiting and killing helicobacter pylori, which comprises 0.2-50 wt% of antibacterial peptide Nisin, 20-70 wt% of citric acid and 5-70 wt% of collagen peptide as active ingredients. The invention also provides a preparation method of the antibacterial peptide modifier PEG-Nisin and application of the composition in preparing health products or medicines. The series products such as food, health care products, medicines and the like prepared by the composition can inhibit and kill H.pyri for a long time, do not generate drug-resistant bacteria, and provide nutritional support for patients.

Description

Antibacterial peptide composition for inhibiting and killing helicobacter pylori and preparation method and application thereof
Technical Field
The invention belongs to the technical field of biological medicines, and particularly relates to an antibacterial peptide composition for inhibiting and killing helicobacter pylori, and a preparation method and application thereof.
Background
Helicobacter pylori (h. pylori), to which global relatives have conducted extensive clinical, laboratory and epidemiological studies or investigations since its first discovery in australia Warren and Marshal in 1983, was assigned by the world health organization international agency for research on cancer (WTO/IARC) in 1994 as a class I carcinogen. The related diseases caused by the infection of the H.pyri is long-term chronic diseases, and the early detection and eradication of the H.pyri is the key for reducing and reducing the incidence rate of the diseases related to the infection of the H.pyri.
Epidemiological investigations have shown that more than half of the people worldwide are infected with h.pyri, and that the infection rate of children and adults with h.pyri is much higher in developing countries than in developed countries. Children are the most at risk group affected by h. Early control of infection rates and increased eradication rates have become a serious problem in developing the public health care industry in China. Shown by gastrointestinal disease tissue (WGO) data, the drug resistance rate of H.pyrori to various clinically used antibiotics is higher, and the drug resistance rate to nitroformazan is 76%, the drug resistance rate of clarithromycin is 27-28%, and the drug resistance rate of amoxicillin is 3%. The high drug resistance of h.pyri leads to a decrease in the eradication rate of h.pyri, which increases the difficulty of clinical eradication of h.pyri.
Nisin is a polypeptide secreted by lactococcus lactis, a peptide with antibacterial activity consisting of 34 amino acid residues, having a molecular weight of 3510, usually present in the form of a dimer. In the natural state, Nisin has two forms of Nisin A and Nisin Z, and the bacteriostatic ability of the Nisin is stronger than that of the Nisin Z. Nisin is thought to have an effect on both vegetative cells and spores of bacteria, and its main point of action on vegetative cells is the cytoplasmic membrane, inhibiting the biosynthesis of peptidoglycan in the cell wall, thereby hindering the synthesis of the cell wall plasma membrane and phospholipid compounds, causing the leakage of intracellular material, resulting in cell lysis. The effect on the spores is to inhibit their germination during the initial stages of their expansion. Nisin is widely used in more than 60 countries, mainly as a natural preservative for dairy products, meat products, canned foods and beverages. Compared with other preservatives, Nisin has the advantages of safety, no toxicity, strong antibacterial property, good water solubility, good thermal stability and the like, but Nisin also has the defect of narrow antibacterial spectrum. Bacteriocins can be adsorbed on bacteria-producing cells under the condition of pH 6.0-6.5, the cells are collected by centrifugation, and then the bacteriocins are dissociated from the cells under the condition of pH 2. A.Guiotto and Y.Imura et al have tried to respectively modify the antimicrobial peptides nisin A and tachplesin I with 5kDa PEG (references A.Guiotto, et al., IlFarmaco 2003, 58, 45-50 and Y.Imura, et al., Biochim.Biophys.acta 2007, 1768, 1160-1169) to optimize the antibacterial activity, but the results show that the activity of the antimicrobial peptides is sharply reduced or even completely inactivated after being modified with PEG. How to make the antibacterial peptide have the degradation effect of resisting proteolytic enzyme and retain the equivalent antibacterial activity is an important problem to be solved at present.
Disclosure of Invention
Aiming at the situation that helicobacter pylori has higher drug resistance to conventional therapeutic drugs and the technical problem of optimizing the activity of the antibacterial peptide in the prior art, the invention aims to provide an antibacterial peptide composition which has no drug resistance and can obviously inhibit the helicobacter pylori and even eradicate the helicobacter pylori in a patient.
The antibacterial peptide composition provided by the invention is specifically as follows:
a composition for inhibiting and killing helicobacter pylori comprises 0.2-50 wt% of antibacterial peptide Nisin, 20-70 wt% of citric acid and 5-70 wt% of collagen peptide as active ingredients.
In one embodiment according to the present invention, the antimicrobial peptide Nisin is antimicrobial peptide Nisin a or Z.
In one embodiment according to the present invention, the antibacterial peptide Nisin is PEG-Nisin modified by PEG selected from PEG200, PEG400, PEG600, PEG800 or PEG 1000.
The invention also provides application of the composition in preparing a medicament or health-care product for relieving or treating helicobacter pylori infection.
The invention further provides a preparation method of the antibacterial peptide modifier PEG-Nisin, which comprises the following steps:
1) reacting antibacterial peptide Nisin with thionyl chloride to obtain a Nisin-chloroformyl compound;
2) dropwise adding an anhydrous benzene solution of PGE into the Nisin-chloroformyl compound, then slowly adding triethylamine, and heating and refluxing to react in an inert atmosphere until the reaction is complete;
3) filtering, concentrating the filtrate, and precipitating with anhydrous ether to obtain antibacterial peptide modifier PEG-Nisin.
In one embodiment of the invention, the dosage ratio of the antibacterial peptide Nisin to the thionyl chloride is 1: 3.
in one embodiment according to the invention, the ratio of the total amount of the compound in g: mL: and the dosage ratio of the PEG, the Nisin-chloroformyl compound and the triethylamine is 20-100: 60: 7.
the antibacterial peptide composition for inhibiting and killing helicobacter pylori provided by the invention has the following beneficial effects:
1. the series products such as food, health care products, medicines and the like prepared by the composition can inhibit and kill over 95 percent of H.pyri, thereby reducing the possibility of diseases caused by H.pyri infection.
2. The series products such as food, health care products, medicines and the like prepared by the composition can inhibit and kill H.pyri for a long time, do not generate drug-resistant bacteria, and provide nutritional support for patients.
Drawings
FIG. 1 is a graph of the bactericidal profile of an antimicrobial peptide composition according to the present invention;
fig. 2 is a schematic diagram showing the results of an antibacterial experiment in which collagen and antibacterial peptide act on h.pyroris together;
FIG. 3 is a graph showing the results of a bacteriostatic loop experiment, wherein A is a blank control and B is an antimicrobial peptide composition applied according to the present invention;
FIG. 4 is a graph showing the results of the experiment of killing helicobacter pylori in zebra fish by the antibacterial peptide.
Detailed Description
The following examples and figures are provided to further illustrate the present application and are not intended to limit the scope of the present application.
Specific embodiments of the present application will be described in more detail below. These embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the scope of the disclosure to those skilled in the art.
In the following description and in the claims, the terms "include" and "comprise" are used in an open-ended fashion, and thus should be interpreted to mean "include, but not limited to. The description which follows is a preferred embodiment of the present application, but is made for the purpose of illustrating the general principles of the application and not for the purpose of limiting the scope of the application. The protection scope of the present application shall be subject to the definitions of the appended claims.
Unless otherwise specified, the reagents used in the present invention are commercially available.
Example 1 Synthesis of PEG2000-Nisin
1. Synthesis of Nisin-chloroformyl compound
Taking 3.0g of Nisin, adding 9ml of refined thionyl chloride into a 50ml glass bottle, slowly dropwise adding under magnetic stirring, carrying out oil bath at 70 ℃ after dropwise adding is finished, reacting for 1 hour, wherein bubbles are generated in the reaction, and stopping the reaction when no solid exists in the reaction mixture and no bubbles are generated any more. And (3) evaporating excessive thionyl chloride under reduced pressure, adding dry benzene, boiling for several minutes, evaporating the benzene under reduced pressure, and evaporating to dryness to obtain a white solid, namely the Nisin-chloroformyl compound.
2. Synthesis of Nisin modified by polyethylene glycol
PEG 20010 g was taken out, and 30ml of anhydrous benzene was added to a 100ml glass bottle and dissolved under magnetic stirring. Dropwise adding the dissolved polyethylene glycol into chloroformyl chloride, slowly adding 3.5ml of triethylamine, magnetically stirring under nitrogen atmosphere, refluxing in an oil bath at 80 ℃ for 24h, filtering out a precipitate after the reaction is finished, concentrating the filtrate, and repeatedly precipitating with anhydrous ether for three times to obtain a refined product.
Table 1 shows the amount composition of the prepared crosslinked material
Sample (I) Nisin(g) Thionyl chloride (ml) PEG(g)
1 3.0 9.0 10.0(PEG200)
2 3.0 9.0 20.0(PEG400)
3 3.0 9.0 30.0(PEG600)
4 3.0 9.0 40.0(PEG800)
5 3.0 9.0 50.0(PEG1000)
Example 2 bacteriostatic experiments against h
Nisin-PEG400 was diluted in sterile test tubes with 1ml h. pyris medium to a series of concentration gradients, in order 0, 1, 2, 8, 16, 32, 128 MIC. 1ml of bacterial suspension (2.0X 10) was added to each tube in sequence6CFU/ml), the concentration of the product in each test tube is 0, 0.5, 1, 4, 8, 16 and 64MIC in sequence, and the final concentration of the bacterial liquid is about 1.0 multiplied by 106CFU/ml. After fully mixing, placing the mixture in a microaerophilic environment for culturing for 48 to 72 hours. Another tube was filled with 1ml h. pyris and 1ml sterile saline as a blank. According to a set time point0. 8, 16, 24, 32, 40, 48, 56, 64 and 72 hours, taking 100 mul of culture, performing gradient dilution, uniformly coating on a flat plate, placing the flat plate in a microaerophilic incubator at 37 ℃ for incubation for 72 hours, and selecting the flat plate with the colony number of 30-300 for viable colony counting. The sterilization curve of Nisin-PEG400 against h.pyris was plotted as time (x) and the logarithm of the number of bacteria (y) corresponding to that time.
As shown in FIG. 1, the results are obtained from the sterilization curve, and when the concentration is 1MIC or more, the time taken to kill all bacteria is gradually shortened as the concentration increases, and the curve is remarkably shifted to the left. Indicating that the increased concentration resulted in an increased rate of sterilization. When the Nisin-PEG400 concentration is 0.5MIC, a certain bacteriostasis effect can be maintained.
Example 3 bacteriostatic experiment of H.pyroris by co-action of collagen and antibacterial peptide
The antibacterial peptide Nisin-PEG 40010 mg, citric acid 500mg and collagen peptide 490mg were co-dissolved in H.pyroris medium to form a composition stock solution.
Taking a sterile test tube, and diluting the composition stock solution into a series of concentration gradients of 0, 2 and 8MIC in sequence by using 1ml of H. 1ml of bacterial suspension (2.0X 10) was added to each tube in sequence6CFU/ml), the concentration of the product in each test tube is 0, 1 and 4MIC in sequence, and the final concentration of the bacterial liquid is about 1.0 multiplied by 106CFU/ml. After fully mixing, placing the mixture in a microaerophilic environment for culturing for 48 to 72 hours. Another tube was filled with 1ml h. pyris and 1ml sterile saline as a blank. According to the set time points of 0, 8, 16, 24, 32, 40, 48, 56, 64 and 72h, 100 mu l of culture is taken to be diluted in a gradient way and then evenly coated on a flat plate, the flat plate is placed in a microaerophilic incubator at 37 ℃ for incubation for 72h, and the flat plate with the colony number of 30-300 is selected for counting viable bacteria colonies. The sterilization curve of the composition against h.pyroris is plotted as time (x) and the logarithm of the number of bacteria (y) corresponding to the time. As shown in fig. 2, the bactericidal effect of the collagen-containing composition is better than that of the composition without collagen, and it can be seen that the addition of collagen helps to maintain the activity of the antimicrobial peptide.
Example 4
Selecting one or more of antibacterial peptide Nisn or antibacterial peptide Nisin modified by PEG200, 400, 600, 800 and 1000 with the daily oral dose of 0.5g-5g to be combined with citric acid or salt thereof and collagen peptide for administration, and taking a postprandial oral mode, wherein the mass ratio of the main components is 1: 2: 7.
experimental example 5
Selecting 0.5g-5g of antibacterial peptide Nisn or one or more of PEG200, 400, 600, 800 and 1000 modified antibacterial peptide Nisin combined with citric acid or salt thereof, 1/10 of collagen peptide combined dose (mass ratio of main components is 1: 2: 7) dissolved in ddH2In O, laboratory-stage verification of helicobacter pylori inhibition was carried out according to standard procedures and culture, and the results showed that it had a strong inhibitory effect on helicobacter pylori.
Example 6 bacteriostatic Ring test
1. Experimental Material
Helicobacter pylori (ATCC43504) suspension.
Bacteriostatic agent carrier (circular Xinhua No. one qualitative filter paper sheet with diameter of 5mm, pressure steam sterilizing, oven drying at 120 deg.C for 2h, and storing for use).
Micropipette (5-50. mu.l, adjustable).
A vernier caliper.
Nutrient agar culture medium, tryptone soybean agar culture medium and sandcastle agar culture medium.
2. Experimental procedure
(1) Preparing the bacteriostatic tablets:
a sterile and dried filter paper sheet was taken, 20. mu.l of the composition stock solution prepared according to example 3 was dropped on each sheet, and then the filter paper sheet was placed flat in a clean sterile plate, and was left to open and put in an incubator (37 ℃ C.) for baking or was left to dry naturally at room temperature for later use. The dissoluble antibacterial product can be directly made into round pieces (blocks) with the diameter of 5mm and the thickness of not more than 4mm, and every 4 round pieces (blocks) are in a group.
(2) Preparation of negative control sample:
taking a sterile dry filter paper sheet, dripping 20 mu l of sterile distilled water into each sheet, and drying for later use.
(3) Inoculation of test bacteria:
using sterile cotton swabThe dipping concentration is 5 × 105cfu/ml～5×106cfu/ml test bacterial suspension was evenly spread on the surface of nutrient agar medium plate for 3 times. For each application 1 time, the plate should be rotated 60 °, and finally the cotton swab is applied around the edge of the plate for one revolution. The plate was covered and dried at room temperature for 5 min.
(4) Sticking bacteriostatic agent sample pieces:
each test is stuck with 1 infectious bacterium plate, and each plate is stuck with 4 test sample plates and 1 negative control sample plate, and the number of the negative control sample plates is 5. A sample was taken with sterile forceps and placed on the surface of the plate. The distance between the centers of the various pieces is more than 25mm, and the distance between the centers of the various pieces and the periphery of the flat plate is more than 15 mm. Placing the mixture in a 37 ℃ incubator, culturing for 16-18 h and observing the result.
The diameter of the antibacterial ring was measured with a vernier caliper and recorded. The experiment was repeated 3 times.
(5) And (4) evaluating the results:
a) judging the bacteriostatic action:
if the diameter of the bacteriostatic ring is larger than 7mm, the bacteriostatic action is judged, and if the diameter of the bacteriostatic ring is smaller than or equal to 7mm, the bacteriostatic action is judged not.
b) And 3 times of repeated tests show that the product has the bacteriostatic effect, and the product is judged to be qualified.
c) The negative control group should have no bacteriostatic ring, otherwise the test is invalid.
3. Results of the experiment
The results are shown in fig. 3, and the statistical results of the diameters of the inhibition zones are shown in the following table:
bacterial strain Control group Diameter of drug zone of inhibition
Helicobacter pylori
0 8.2±0.6
The antibacterial peptide composition provided by the invention has an obvious inhibiting effect on helicobacter pylori when the concentration is 0.05 mu g/ml.
Example 7 pharmaceutical effect experiment for inhibiting and killing helicobacter pylori based on zebra fish model
1. Experimental Material
AB wild type zebra fish, helicobacter pylori, FITC-d-LYS dye
2. The experimental steps are as follows:
1) selecting healthy young fishes which are hatched for 96 hours;
2) diluting the dye stock solution with DMSO, adding 5 μ L of dye stock solution into per ml of bacterial solution during experiment until the concentration of the dye stock solution reaches 0.1mM, and culturing the bacterial solution in a 37 deg.C incubator in the dark until the OD of the bacterial solution is measured6001-1.5, centrifuging the bacterial liquid, washing with LB culture solution for three times, then washing with PBS culture solution for three times, and observing whether the bacteria are stained or not under a body type microscope;
3) feeding a bacterial liquid with a proper concentration into a zebra fish digestive system in a feeding mode and observing whether the molding is successful or not;
4) the zebra fish successfully molded is put into antibacterial peptide bacteriostatic solution (prepared according to example 3) with different concentrations to continue to observe whether Hp carrying fluorescence is completely inhibited and killed in the zebra fish body.
As shown in the experimental result of FIG. 4, when the concentration of the antibacterial peptide composition provided by the invention is 0.4 mug/ml, the Hp in the zebra fish body can be completely clear after the action time is 24 hours.
Through a comparison experiment of 30 zebra fish in each group, the gastrointestinal tract Hp of the zebra fish fed with the test sample group is obviously changed compared with a normal control group, and the fluorescence is completely disappeared.
The experiments prove that the antibacterial peptide composition provided by the invention has a remarkable effect of inhibiting and killing helicobacter pylori (Hp) in a living animal body.
It should be noted that the above examples are only used to illustrate the technical solutions of the present invention and not to limit them. Although the present invention has been described in detail with reference to the examples given, those skilled in the art can modify the technical solution of the present invention as needed or equivalent substitutions without departing from the spirit and scope of the technical solution of the present invention.

Claims (7)

1. A composition for inhibiting and killing helicobacter pylori is characterized in that active ingredients comprise 0.2-50 wt% of antibacterial peptide Nisin, 20-70 wt% of citric acid and 5-70 wt% of collagen peptide.
2. The composition of claim 1, wherein the antimicrobial peptide Nisin is antimicrobial peptide Nisin a or Nisin Z.
3. The composition of claim 1, wherein the antibacterial peptide Nisin is PEG-Nisin modified by PEG, and the PEG is selected from PEG200, PEG400, PEG600, PEG800 or PEG 1000.
4. Use of a composition according to any one of claims 1 to 3 in the manufacture of a medicament or health product for the alleviation or treatment of helicobacter pylori infection.
5. A preparation method of an antibacterial peptide modifier PEG-Nisin is characterized by comprising the following steps:
1) reacting antibacterial peptide Nisin with thionyl chloride to obtain a Nisin-chloroformyl compound;
2) dropwise adding an anhydrous benzene solution of PGE into the Nisin-chloroformyl compound, then slowly adding triethylamine, and heating and refluxing to react in an inert atmosphere until the reaction is complete;
3) filtering, concentrating the filtrate, and precipitating with anhydrous ether to obtain antibacterial peptide modifier PEG-Nisin.
6. The preparation method according to claim 5, wherein the dosage ratio of the antibacterial peptide Nisin to the thionyl chloride is 1: 3.
7. the method of claim 5, wherein the ratio of the total amount of the compound in g: mL: and the dosage ratio of the PEG, the Nisin-chloroformyl compound and the triethylamine is 20-100: 60: 7.
CN202111527966.7A 2021-12-14 2021-12-14 Antibacterial peptide composition for inhibiting and killing helicobacter pylori and preparation method and application thereof Pending CN114099640A (en)

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