CN113041340A - Biological sterilization gel and preparation method thereof - Google Patents

Biological sterilization gel and preparation method thereof Download PDF

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CN113041340A
CN113041340A CN201911387213.3A CN201911387213A CN113041340A CN 113041340 A CN113041340 A CN 113041340A CN 201911387213 A CN201911387213 A CN 201911387213A CN 113041340 A CN113041340 A CN 113041340A
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percent
preparation
components
antibacterial peptide
solution
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叶贵子
黄青山
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Kunshan Biogreen 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
    • A61K38/04—Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
    • A61K38/10—Peptides having 12 to 20 amino acids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/74—Synthetic polymeric materials
    • A61K31/785—Polymers containing nitrogen
    • 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
    • A61K47/02—Inorganic compounds
    • 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
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/08—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing oxygen, e.g. ethers, acetals, ketones, quinones, aldehydes, peroxides
    • A61K47/10—Alcohols; Phenols; Salts thereof, e.g. glycerol; Polyethylene glycols [PEG]; Poloxamers; PEG/POE alkyl ethers
    • 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
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/16—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing nitrogen, e.g. nitro-, nitroso-, azo-compounds, nitriles, cyanates
    • A61K47/18—Amines; Amides; Ureas; Quaternary ammonium compounds; Amino acids; Oligopeptides having up to five amino acids
    • 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
    • A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
    • A61K47/32—Macromolecular compounds obtained by reactions only involving carbon-to-carbon unsaturated bonds, e.g. carbomers, poly(meth)acrylates, or polyvinyl pyrrolidone
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/06—Ointments; Bases therefor; Other semi-solid forms, e.g. creams, sticks, gels
    • 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

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  • General Chemical & Material Sciences (AREA)
  • Inorganic Chemistry (AREA)
  • Oil, Petroleum & Natural Gas (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
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  • Organic Chemistry (AREA)
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Abstract

The invention discloses a biological sterilization gel and a preparation method thereof, the preparation comprises antibacterial peptide K17 and polyhexamethylene biguanide (PHMB), and the components and the weight percentage content are as follows: 170.1 to 1 percent of antibacterial peptide K; 0.05 to 5 percent of polyhexamethylene biguanide; 0.1% -10% of carbomer; 0.1 to 5 percent of N-acetyl-D-amino; 0.5 to 2 percent of sodium chloride; 0.5-30% of glycerin; the rest is distilled water; the sum of the weight percentages of the components is 100 percent. The antibacterial peptide gel preparation overcomes the defects of strong irritation, easy generation of drug resistance and ineffective drug-resistant bacterial infection of the existing antibacterial preparation, is convenient to use, has strong activity exerting action time and obvious sterilization effect.

Description

Biological sterilization gel and preparation method thereof
Technical Field
The invention belongs to the technical field of biology, and relates to a biological sterilization gel and a preparation method thereof.
Background
Currently, the clinical pathogen infection is quite serious, and it has become a big killer threatening the health of human beings in recent 20 years. Moreover, more and more bacteria are resistant to antibiotic treatment, thereby posing a serious threat to public health. In the united states, 1998, more life is lost due to drug-resistant bacterial infections than on the vietnam battlefield. Pathogens that can be killed by antibiotics at once with ease such as: staphylococcus aureus, pneumococcus, pseudomonas aeruginosa, candida albicans, escherichia coli and the like are increasingly fierce due to drug resistance, and people are increasingly stranded. The drug-resistant staphylococcus aureus is the most active molecule, and the infection caused by the drug-resistant staphylococcus aureus not only has poor or completely ineffective curative effect under conventional treatment, but also can change the ecological condition of normal flora of patients, thereby causing additional pain to the patients, and increasing economic burden and unfortunate property.
In the face of increasingly serious pathogenic infection, chemical therapeutic agents, mainly antibiotics, are clinically and massively applied at present, but with the wide use of antibiotics, a plurality of adverse factors continuously appear, on one hand, the drug-resistant bacteria which appear along with the antibiotics are more and more, and the elimination of the drug-resistant bacteria becomes more and more difficult; on the other hand, the side effect caused by antibiotics is quite serious, and certain harm is caused to the body and mind of a patient. Therefore, the medical field is always searching for a bactericidal preparation which can replace antibiotics, can play a bactericidal effect and is not easy to generate drug-resistant strains.
Disclosure of Invention
The invention aims to overcome the defects that the existing antibacterial preparation has strong irritation, is easy to generate drug resistance and is ineffective to drug-resistant bacterial infection, and provides a biological sterilization gel which is convenient to use, strong in activity exertion action time and obvious in sterilization effect.
The invention also provides a preparation method of the biological bactericidal gel.
The technical scheme of the invention is as follows:
a biological sterilization gel comprises the following components in percentage by weight:
Figure BSA0000198567750000011
the sum of the weight percentages of the components is 100 percent.
The preferable components and weight percentage are as follows:
Figure BSA0000198567750000012
Figure BSA0000198567750000021
the sum of the weight percentages of the components is 100 percent.
More preferred components and weight percentages are:
Figure BSA0000198567750000022
the sum of the weight percentages of the components is 100 percent.
The biological antibacterial peptide K17 is a polypeptide with an amino acid sequence of Lys Trp Lys Ser Phe Ile Lys Lys Leu Thr Ser Lys Phe Leu His Ser Lys Lys Lys Phe, and the preparation, purification and properties of the biological antibacterial peptide K17 are described in patent of invention No. CN 101570569B.
The antibacterial peptide K17 and polyhexamethylene biguanide (PHMB) or salt thereof form a soluble complex, and the addition ratio of the antibacterial peptide K17 to the polyhexamethylene biguanide (PHMB) is 1: 0.5-1: 5.
The action mechanism of the antibacterial peptide K17 is to act on cell membranes, destroy the integrity of the cell membranes and cause the leakage of cell contents to die. The PHMB sterilization process is a physical action, can quickly adsorb and capture bacteria or viruses, and forms polymers on the surface of microorganisms to inhibit the division and replication of the bacteria and the viruses. The inventor analyzes the action characteristics of the two and organically combines the two, thereby achieving the sterilizing effect which is obviously higher than that of the original single use: according to the formula and the method, firstly, the antibacterial peptide K17 and the PHMB form a high-molecular complex, so that on one hand, the antibacterial peptide K17 protein is protected, on the other hand, microorganisms are rapidly gathered and captured under the action of the PHMB, the binding efficiency of the antibacterial peptide K17 and bacterial cells is improved by exponential multiple, and the microorganisms are rapidly and efficiently killed under the combined action of the antibacterial peptide K17 and the PHMB. The antibacterial composition provided by the invention has the advantages that the sterilization rate is obviously improved, the use effective concentration can be lower, the biological safety is higher, the use cost is lower, and the antibacterial composition can be better applied to the antibacterial field.
The N-acetyl-D-amino plays a role in stabilizing protein and enhancing activity, can coat protein or polypeptide and is free from being damaged by other substances, so that the stability of the protein or the polypeptide is enhanced;
the carbomer is a high-molecular polymer of acrylic acid bonded allyl sucrose or pentaerythritol allyl ether, and has strong hygroscopicity. The molecular has a large number of carboxylic acid groups, and the carboxylic acid groups rapidly swell in water to form gel, and have certain strength and elasticity. Carbomer as a novel polymer dressing has wide application, can be used for preparing hydrogel, and can be used as suspending agent, auxiliary emulsifier, etc.
The glycerin has the effects of lubrication and freeze protection, can prolong the local curative effect of an agent on mucous membrane, has good lubrication and skin protection effects, and can improve the application range of the product;
the sodium chloride of the invention is an ionic compound, plays a role in regulating the osmotic pressure of the preparation, assists in enhancing the stability of the preparation, and is suitable for wound surfaces or mucous membranes of different parts.
A method for preparing a formulation of the invention, the method comprising the steps of:
(1) weighing antibacterial peptide K17 and polyhexamethylene biguanide (PHMB) according to a certain proportion, mixing and dissolving in water with pH of 6.5-7.0 at 15-35 deg.C, stirring at 50-150rpm, and standing to obtain solution A.
(2) Respectively weighing sodium chloride, N-acetyl-D-amino and carbomer according to the proportion, preparing a solution B, and uniformly stirring;
(3) weighing glycerol according to the proportion, stirring the solution B, slowly adding the solution A and the glycerol into the solution B, and uniformly stirring to obtain a finished product.
Compared with the prior art, the invention has the beneficial effects that:
(1) the invention adopts antibacterial peptide K17 as a main raw material, and regulator such as carbomer, N-acetyl-D-amino, glycerol, sodium chloride and the like are used as auxiliary materials, and the synergistic effect among the raw materials and the auxiliary materials is utilized to prepare the biological antibacterial gel with high antibacterial efficiency, broad spectrum, mild action and good biocompatibility.
(2) The biological antibacterial gel of the invention has biocompatibility meeting the GB/T16886.11-2011 requirement.
(3) Is suitable for wound surface or mucosa infection caused by various bacteria pathogenic bacteria.
(4) Has no toxic and side effects on organisms. The core antibacterial components are protein products, which can stimulate skin and mucosa, do not generate any toxic or side effect on organisms, can be completely degraded through metabolism, have no residue, and can not be enriched.
Detailed Description
The present invention will be described in further detail with reference to specific embodiments.
Example 1
1. The components and the proportion are as follows: (weight percent)
Figure BSA0000198567750000031
The sum of the weight percentages of the components is 100 percent.
2. Preparation step
(1) The antibacterial peptide K17 and the polyhexamethylene biguanide (PHMB) are respectively weighed according to the proportion, mixed and dissolved in water with the pH value of 6.5 at the temperature of 15 ℃, stirred evenly at the speed of 150rpm and kept stand to obtain solution A.
(2) Respectively weighing sodium chloride, N-acetyl-D-amino and carbomer according to the proportion, preparing a solution B, and uniformly stirring;
(3) weighing glycerol according to the proportion, stirring the solution B, slowly adding the solution A and the glycerol into the solution B, and uniformly stirring to obtain a finished product.
Example 2
1. The components and the proportion are as follows: (weight percent)
Figure BSA0000198567750000032
Figure BSA0000198567750000041
The sum of the weight percentages of the components is 100 percent.
2. Preparation step
(1) The antibacterial peptide K17 and the polyhexamethylene biguanide (PHMB) are respectively weighed according to the proportion, mixed and dissolved in water with the pH value of 7.0 at the temperature of 35 ℃, stirred evenly at the speed of 50rpm and kept stand to obtain a solution A.
(2) Respectively weighing sodium chloride, N-acetyl-D-amino and carbomer according to the proportion, preparing a solution B, and uniformly stirring;
(3) weighing glycerol according to the proportion, stirring the solution B, slowly adding the solution A and the glycerol into the solution B, and uniformly stirring to obtain a finished product.
Example 3
1. The components and the proportion are as follows: (weight percent)
Figure BSA0000198567750000042
The sum of the weight percentages of the components is 100 percent.
2. Preparation step
(1) The antibacterial peptide K17 and the polyhexamethylene biguanide (PHMB) are respectively weighed according to the proportion, mixed and dissolved in water with the pH value of 6.8 at the temperature of 25 ℃, stirred evenly at the speed of 100rpm and kept stand to obtain solution A.
(2) Respectively weighing sodium chloride, N-acetyl-D-amino and carbomer according to the proportion, preparing a solution B, and uniformly stirring;
(3) weighing glycerol according to the proportion, stirring the solution B, slowly adding the solution A and the glycerol into the solution B, and uniformly stirring to obtain a finished product.
Example 4
1. The components and the proportion are as follows: (weight percent)
Figure BSA0000198567750000043
The sum of the weight percentages of the components is 100 percent.
2. Preparation step
(1) The antibacterial peptide K17 and the polyhexamethylene biguanide (PHMB) are respectively weighed according to the proportion, mixed and dissolved in water with the pH value of 6.5 at the temperature of 25 ℃, stirred evenly at the speed of 150rpm and kept stand to obtain solution A.
(2) Respectively weighing sodium chloride, N-acetyl-D-amino and carbomer according to the proportion, preparing a solution B, and uniformly stirring;
(3) weighing glycerol according to the proportion, stirring the solution B, slowly adding the solution A and the glycerol into the solution B, and uniformly stirring to obtain a finished product.
Example 5
1. The components and the proportion are as follows: (weight percent)
Figure BSA0000198567750000051
The sum of the weight percentages of the components is 100 percent.
2. Preparation step
(1) The antibacterial peptide K17 and the polyhexamethylene biguanide (PHMB) are respectively weighed according to the proportion, mixed and dissolved in water with the temperature of 30 ℃ and the pH value of 6.5, stirred evenly at 150rpm and kept stand to obtain solution A.
(2) Respectively weighing sodium chloride, N-acetyl-D-amino and carbomer according to the proportion, preparing a solution B, and uniformly stirring;
(3) weighing glycerol according to the proportion, stirring the solution B, slowly adding the solution A and the glycerol into the solution B, and uniformly stirring to obtain a finished product.
Example 6
The clinical trial report of the biocidal gel of example 1 demonstrates its efficacy:
the first clinical test is as follows: sterilization experiment of biological sterilization gel on staphylococcus aureus, escherichia coli and candida albicans
First, equipment
1. Test bacteria: staphylococcus aureus generation 6 (provided by Jun medical academy of sciences with flowing billy), Escherichia coli generation 6 (provided by Jun medical academy of sciences disinfection and detection center), and Candida albicans generation 6 (provided by Jun medical academy with flowing billy detection center)
2. Sample preparation: biobactericidal gel example 1
3. Neutralizing agent: D/E broth of 2% histidine, 8% lecithin wax, 10% tween-80
4. PBS: 0.03mol/L phosphate buffer solution
5. GNP-9160 type water-proof constant temperature incubator (Shanghai Jing hong experiment equipment Co., Ltd.)
6. Culture medium: common nutrient agar
7. Mechanical stopwatch (504 type)
8. Adjustable constant temperature water bath box (CBN28-3)
Second, method
1. The inspection basis is as follows: sanitary Standard for Disposable sanitary articles GB15979-2002 appendix C3
2. And (3) identification test of a neutralizer: test strains: concentration of staphylococcus aureus sample: 1: 20 diluent, action time: 0.5min
3. Quantitative sterilization test: sample concentration: 1: 20 dilution
4. The test temperature is 20 ℃, and the test is repeated for 3 times
Three, result in
When the test temperature is 20 ℃, the sterilization rate of the sample on staphylococcus aureus and escherichia coli is 100% after the sample is acted by the diluent with the ratio of 1: 2 for 2 minutes, 5 minutes, 10 minutes and 20 minutes.
TABLE 1 Sterilization effect on the experimental bacteria
Figure BSA0000198567750000061
Example 7
EXAMPLE 2 vaginal Mucosa irritation test with Biobactericidal gel
Materials and animals
1. The test substance: the sample is colorless liquid, and the original sample liquid is taken as a test substance.
2. Animals: 6 healthy female New Zealand white rabbits, the common grade, provided in the Jinling breeding rabbit farm, the certification number: SCXK (Su) 2012-0003, weight 2.0-2.4 kg.
3. The test conditions are as follows: animal environmental facility certification number: SYXK (Su) No. 2012-0036; ambient temperature: 22 +/-2 ℃; relative humidity: 40-70 percent; feed source and identification number: jiangsu province, cooperative pharmaceutical bioengineering, Inc., Su A gao Sheng (2002) 009. The main apparatus is as follows: infant scales (05-324), pathological microtomes, microscopes.
Second, method
1. The inspection basis is as follows: the second part of the "Disinfection Specification" of the Ministry of health (2002) 2.3.5 vaginal mucosa irritation test.
2. The test method comprises the following steps: animals were divided into test and control groups of 3 animals each. The animal is fixed on the back, and the perineum and the vaginal orifice are exposed. A small size catheter was connected to a 2mL syringe, moistened with the test substance, gently inserted into the animal's vagina (4-5cm), slowly injected into 2mL of the test substance, and the catheter was withdrawn to complete the staining. normal saline of animals in a control group is treated in the same way, the animals are killed by an air embolism method after 24mL, the complete vagina is taken out after the abdominal incision, the longitudinal incision is carried out, whether congestion, edema and other manifestations exist is observed by naked eyes, the vagina is placed into 10% formalin for fixing for more than 24h for reference when pathological material drawing is carried out, tissues at two ends and 3 parts in the center of the vagina are selected for flaking, and histopathological examination is carried out after HE staining.
Third, experimental results
The mild moderate edema of the vaginal mucosa of each animal in the control group No. 2 and the test group can be seen by visual observation. Histopathological observations are shown in the table below.
TABLE 2 test substance response to vaginal mucosal irritation in animals
Figure BSA0000198567750000071
Fourth, conclusion
The stimulation index of the tested substance to the rabbit vaginal mucosa is 1.56, and the stimulation intensity of the vaginal mucosa is extremely light stimulation.

Claims (2)

1. The biological sterilization gel is characterized by comprising the following components in percentage by weight:
Figure FSA0000198567740000011
the sum of the weight percentages of the components is 100 percent.
2. The preferred composition and weight percent as claimed in claim 1 is:
Figure FSA0000198567740000012
the sum of the weight percentages of the components is 100 percent.
CN201911387213.3A 2019-12-27 2019-12-27 Biological sterilization gel and preparation method thereof Pending CN113041340A (en)

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Application Number Priority Date Filing Date Title
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Country Status (1)

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