CN113081977B - Anti-infection antibacterial peptide freeze-dried preparation and preparation method thereof - Google Patents
Anti-infection antibacterial peptide freeze-dried preparation and preparation method thereof Download PDFInfo
- Publication number
- CN113081977B CN113081977B CN202110483679.4A CN202110483679A CN113081977B CN 113081977 B CN113081977 B CN 113081977B CN 202110483679 A CN202110483679 A CN 202110483679A CN 113081977 B CN113081977 B CN 113081977B
- Authority
- CN
- China
- Prior art keywords
- antibacterial peptide
- freeze
- preparation
- hours
- parts
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Classifications
-
- 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
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/1767—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from invertebrates
-
- 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
- A61K47/08—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing oxygen, e.g. ethers, acetals, ketones, quinones, aldehydes, peroxides
- A61K47/12—Carboxylic acids; Salts or anhydrides thereof
-
- 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
- A61K47/26—Carbohydrates, e.g. sugar alcohols, amino sugars, nucleic acids, mono-, di- or oligo-saccharides; Derivatives thereof, e.g. polysorbates, sorbitan fatty acid esters or glycyrrhizin
-
- 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
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0014—Skin, i.e. galenical aspects of topical compositions
-
- 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
- A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
- A61K9/19—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles lyophilised, i.e. freeze-dried, solutions or dispersions
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
- A61P17/02—Drugs for dermatological disorders for treating wounds, ulcers, burns, scars, keloids, or the like
-
- 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
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/10—Antimycotics
-
- 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
- A61P31/12—Antivirals
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P33/00—Antiparasitic agents
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Veterinary Medicine (AREA)
- Pharmacology & Pharmacy (AREA)
- Public Health (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Animal Behavior & Ethology (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Epidemiology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Dermatology (AREA)
- Communicable Diseases (AREA)
- Oncology (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Virology (AREA)
- Gastroenterology & Hepatology (AREA)
- Zoology (AREA)
- Immunology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Tropical Medicine & Parasitology (AREA)
- Biochemistry (AREA)
- Molecular Biology (AREA)
- Medicinal Preparation (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The invention discloses an anti-infection antibacterial peptide freeze-dried preparation and a preparation method thereof. The antibacterial peptide freeze-dried preparation comprises the following components in parts by mass: 0.05-0.5 part of antibacterial peptide, 2-5 parts of lactose, 5-10 parts of mannitol and 0.1-1 part of sodium citrate, and is prepared by a freeze-drying process. The antibacterial peptide freeze-dried preparation is a local external anti-infective medicament, is suitable for various infections caused by pathogenic bacteria, particularly drug-resistant bacteria, and has wide application prospect.
Description
Technical Field
The invention belongs to the technical field of biological medicines, and particularly relates to an anti-infection antibacterial peptide freeze-dried preparation and a preparation method thereof.
Background
With the development of biomedical technology, reports of antibacterial peptides for antibacterial research are increasing. The antibacterial peptide takes cell membranes as main targets, and the cell membranes lose barrier functions through unique characteristics of the antibacterial peptide, so that the antibacterial peptide is the focus of the field of biological medicines.
Medical infection refers to local tissue and systemic inflammatory response caused by invasion of pathogens such as bacteria, viruses, fungi, parasites, etc. into the human body. Infection of local tissues such as skin lacerations, tears, burns, abscesses, and the like can cause infection. The CN111643559A patent discloses that the existing medicines for treating the above infections are in various forms, such as granules or paste, but all have certain disadvantages. Researchers develop a traditional Chinese medicine powder for detoxifying and promoting tissue regeneration, and the traditional Chinese medicine powder is particularly suitable for exposure therapy. The powder is a preparation with maximum dispersion degree in solid dosage form, and has small particle size, easy dispersion, simple preparation process and easy dosage control. CN1795871A discloses a Chinese and western medicine powder for treating oral ulcer, which has the advantages of simple preparation process, good curative effect and the like. CN10797262A discloses a western medicine powder for treating children's oral ulcer. CN106727621A discloses a western medicine powder for treating toothache, which has the advantages of small toxic and side effects, quick response and the like. The active ingredients of the powder are traditional Chinese medicine ingredients or a mixture of western medicine active ingredients, the preparation of the powder is mostly carried out at room temperature, and the raw materials have good stability in the preparation process.
The antibacterial peptide is easy to degrade under the wet and hot conditions, and the antibacterial peptide can generate degradation impurities due to local overhigh temperature generated in the preparation process, so that the product quality is poor. Aiming at a heat-unstable polypeptide product, the stability of the antibacterial peptide can be improved by preparing the antibacterial peptide freeze-dried composition by the freeze-drying process of the antibacterial peptide. CN104043098A discloses an antibacterial peptide freeze-dried preparation for treating vaginitis and a preparation method thereof, wherein the antibacterial peptide freeze-dried preparation is prepared by freeze-drying an antibacterial peptide raw material solution, but special dissolution is usually required to be added clinically. The product process is complex, and brings certain inconvenience to the clinical application, etc. If the stable antibacterial peptide composition freeze-dried preparation can be prepared by freeze-drying, the convenience of clinical medication can be improved.
Disclosure of Invention
The invention provides an antibacterial peptide freeze-dried preparation which is prepared by a freeze-drying process. The antibacterial peptide freeze-dried preparation is used for anti-infection treatment, so as to hopefully solve the increasingly serious drug resistance problem of bacteria and the like and relieve the pain of patients.
Aiming at a heat-labile polypeptide product, the stability of the antibacterial peptide can be improved by preparing the antibacterial peptide freeze-dried composition by the antibacterial peptide freeze-drying process.
The invention discloses an anti-infection antibacterial peptide freeze-dried preparation which comprises the following components in parts by mass: 0.05-0.5 part of antibacterial peptide, 2-5 parts of lactose, 5-10 parts of mannitol and 0.1-1 part of sodium citrate.
Wherein, the antibacterial peptide is polypeptide with the following general formula:
segment A-I-A-II-segment B;
i is selected from any of the following amino acid residues: l-leucine, D-leucine, L-valine, D-valine, L-alanine, D-alanine, glycine, L-serine, D-serine, L-lysine and D-lysine (L-and D-optical isomeric forms of L, A, S, V and K, and G);
II is selected from any amino acid residue as follows: any amino acid residue selected from: l-leucine, D-leucine, L-valine, D-valine, L-alanine, D-alanine, glycine, L-serine, D-serine, L-lysine and D-lysine (L-and D-optical isomeric forms of L, A, S, V and K, and G);
segment A is shown as SEQ ID No: 1, the sequence is as follows: KWKSFLKTFK, respectively;
segment B is as shown in SEQ ID No: 2, the sequence is as follows: KTVLHTALKAISS, respectively;
a represents an alanine residue;
in the general formula, the direction is from the N terminal to the C terminal.
Preferably, the antibacterial peptide is NA L And D-NA L 。
The NA is L Has the sequence shown in SEQ ID No: 3, the amino acid sequence of which is: KWKSFLKTFKSAAKTVLHTALKAISS, respectively;
the D-NA L Has the sequence shown in SEQ ID No: 4, the amino acid sequence of which is: KWKSFLKTFKSAAKTVLHTALKAISS (all amino acids are in the D-configuration except the A at position 13 which is in the L-configuration).
The pH value of the anti-infective antibacterial peptide freeze-dried preparation is 4.0-6.0.
Preferably, the antibacterial peptide freeze-dried preparation for resisting infection comprises the following components in parts by mass: 0.05-0.5 part of antibacterial peptide, 3-3.5 parts of lactose, 6-6.5 parts of mannitol and 0.4-0.5 part of sodium citrate.
The invention also provides a preparation method of the antibacterial peptide freeze-dried preparation.
The raw materials of the freeze-dried preparation of the antibacterial peptide provided by the invention comprise the following substances in percentage by mass and volume (g/100 ml): 0.5-5 per mill of antibacterial peptide, 2-5 per mill of lactose, 5-10 percent of mannitol, 0.1-1 percent of sodium citrate and the balance of water for injection.
The preparation method of the antibacterial peptide freeze-dried preparation provided by the invention comprises the following steps: adding the antibacterial peptide, lactose and mannitol into water for injection, stirring for dissolving, adding sodium citrate to adjust the pH value to 4.0-6.0, and supplementing the water for injection to the full amount; sterilizing and filtering with 0.22 μm filter membrane, subpackaging in penicillin bottles, and lyophilizing in vacuum lyophilizer; and after freeze-drying is finished, filling nitrogen into the box body of the vacuum freeze-drying machine, keeping the pressure of the box body to be-0.015 Mpa-0 MPa for 10-15 minutes, then pressing the plug, taking out the box and rolling the cover.
Wherein the specific lyophilization profile of the lyophilization is as follows:
a pre-freezing stage: and cooling the cold trap to below-50 ℃, and maintaining for 3-4 hours to freeze the sample.
A main sublimation stage: controlling the temperature of the heat conducting oil to rise to-10 ℃ within 1-2 hours, and maintaining for 8-12 hours; controlling the temperature of the heat conducting oil to rise to 0 ℃ within 1-2 hours, and maintaining for 15-25 hours.
And (3) analysis and drying stage: controlling the temperature of the heat conducting oil to rise to 30 ℃ within 1-2 hours, and maintaining for 5-10 hours.
Compared with the prior art, the invention has the following beneficial effects:
because the antibacterial peptide is easy to degrade under the wet and hot conditions, the technology dissolves the antibacterial peptide, adds the stabilizer to prepare the antibacterial peptide freeze-dried preparation, and can improve the stability of the antibacterial peptide by storing the antibacterial peptide freeze-dried preparation in a solid state. In addition, the product aims to develop a local external anti-infective drug, and the prepared antibacterial peptide freeze-dried preparation is convenient to carry and can be directly used, so that the compliance of patients is improved.
Detailed Description
The present invention will be further illustrated with reference to the following specific examples, but the present invention is not limited to the following examples. The method is a conventional method unless otherwise specified. The starting materials are commercially available from the open literature unless otherwise specified.
The sequences of the antibacterial peptides used in the following examples (having the sequences of SEQ ID Nos: 3) can be synthesized artificially.
Example 1: screening of antibacterial peptide freeze-drying protective agent
The antibacterial peptide freeze-drying protective agent can be selected from sucrose, trehalose, dextran, lactose, mannitol, glycine, inositol, meglumine, sorbitol, arginine, histidine, glycerol, polyvinylpyrrolidone, cyclodextrin, hydroxypropyl methyl cellulose and the like, the invention focuses on the consideration of the influence of lactose, mannitol, cyclodextrin and hydroxypropyl methyl cellulose as the freeze-drying protective agent on the quality of the antibacterial peptide freeze-dried preparation (the preparation method of the antibacterial peptide freeze-dried preparation is shown in example 3), the specific formula is shown in table 1, and the result is shown in table 2.
TABLE 1 formulation composition of lyophilized formulations of antibacterial peptides containing different lyoprotectants
TABLE 2 stability test results of lyophilized formulations of antibacterial peptides (at 25 deg.C)
As can be seen from the analysis of the results in tables 1 and 2, the antibacterial peptide content of formula 1 is still higher than 98% after being placed at 25 ℃ for 3 months, while the antibacterial peptide content of formula 2 and formula 3 is lower than 95% under the same conditions, so that mannitol and lactose are selected as the antibacterial peptide freeze-drying protective agent.
Example 2: investigation of Freeze-drying Nitrogen-charging Process
The prescription composition table of the antibacterial peptide freeze-dried preparation is as follows:
antibacterial peptide 0.1g
Mannitol 6.2g
Lactose 3.3g
Sodium citrate 0.4g
Adding water for injection to 100ml
By adopting the prescription, the influence of nitrogen filling (the box body pressure is up to-0.01 Mpa) on the quality of the antibacterial peptide freeze-dried preparation is investigated in the freeze-drying process, and the result is shown in Table 3.
TABLE 3 influence of Nitrogen charging on the quality of lyophilized formulations of antimicrobial peptides (at 25 ℃ C.)
As can be seen from the analysis of the results in Table 3, when the lyophilized preparation of antibacterial peptide is placed at 25 ℃ for 3 months, the content of the antibacterial peptide in the non-nitrogen-filled group is less than 95%, the content of the antibacterial peptide in the nitrogen-filled group is more than 98%, and the product quality of the antibacterial peptide lyophilized preparation can be improved by filling nitrogen into the headspace.
Example 3: antibacterial peptide freeze-dried preparation
Lyophilized formulation of antimicrobial peptide (one):
antibacterial peptide 0.05g
Mannitol 6.25g
Lactose 3.3g
Sodium citrate 0.4g
Adding water for injection to 100ml
The antibacterial peptide lyophilized preparation (II):
antibacterial peptide 0.5g
Mannitol 6.0g
Lactose 3.1g
Sodium citrate 0.4g
Adding water for injection to 100ml
Antibacterial peptide lyophilized preparation (III):
antibacterial peptide 0.1g
Mannitol 6.2g
Lactose 3.3g
Sodium citrate 0.4g
Adding water for injection to 100ml
Antibacterial peptide lyophilized preparation (IV):
antibacterial peptide 0.2g
Mannitol 6.1g
Lactose 3.3g
Sodium citrate 0.4g
Adding water for injection to 100ml
The preparation process comprises the following steps: adding antibacterial peptide, lactose and mannitol into water for injection, stirring for dissolving, adding sodium citrate to adjust pH to 5.0, and adding water for injection to full dose. Sterilizing and filtering with 0.22 μm filter membrane, subpackaging in penicillin bottles, semi-filling in a vacuum freeze-drying machine, and freeze-drying, wherein the specific freeze-drying curve is as follows:
a pre-freezing stage: and cooling the cold trap to below-50 ℃, and maintaining for 3-4 hours to freeze the sample.
A main sublimation stage: controlling the temperature of the heat conducting oil to rise to-10 ℃ within 1-2 hours, and maintaining for 8-12 hours; controlling the temperature of the heat conducting oil to rise to 0 ℃ within 1-2 hours, and maintaining for 15-25 hours.
And (3) analysis and drying stage: controlling the temperature of the heat conducting oil to rise to 30 ℃ within 1-2 hours, and maintaining for 5-10 hours.
Nitrogen filling: and after the analysis and drying are finished, filling nitrogen into the box body of the freeze dryer, keeping the pressure of the box body to be-0.015 MPa-0 MPa for 10-15 minutes, then pressing the plug, taking out the box and rolling the cover.
The quality evaluation method of the antibacterial peptide freeze-dried preparation comprises the following steps:
1) related substances are as follows: performing high performance liquid chromatography (appendix V D of second part of 2010 edition in Chinese pharmacopoeia) test by using octadecylsilane chemically bonded silica as filler, collecting 20 μ L of solution, injecting into liquid chromatograph, and recording chromatogram. Related substances and limits: the total impurities in the area normalization method cannot exceed 3.0%.
2) The content is as follows: measuring by high performance liquid chromatography (0512 of the four-part general regulation of the national pharmacopoeia 2015 edition), precisely measuring a proper amount of the product, adding water for dissolution, quantitatively diluting to prepare a solution containing 0.25mg per 1ml, using the solution as a test solution, precisely measuring 10 mu l, injecting into a liquid chromatograph, and recording a chromatogram; and taking another appropriate amount of antibacterial peptide reference substance (antibacterial peptide bulk drug), and determining by the same method.
The results of stability study of the antimicrobial peptide lyophilized preparation (i) and the antimicrobial peptide lyophilized preparation (ii) are shown in table 4.
TABLE 4 stability test results of lyophilized formulations of antibacterial peptides (at 4 ℃ C.)
As shown in Table 4, the antibacterial peptide content of both the lyophilized preparation (I) and the lyophilized preparation (II) can be maintained at more than 98% within 12 months at 4 ℃.
Example 4: skin infection resistance test of antibacterial peptide lyophilized preparation
ICR mice, 20 ± 2g, male. Randomly grouped by weight. Each group comprises 10 animals and 5 groups, and is divided into a model group, a lyophilized preparation matrix control group, a levofloxacin gel positive control group, an antibacterial peptide lyophilized preparation (I) group and an antibacterial peptide lyophilized preparation (II) group. Feeding in a common animal house, feeding with common feed, freely taking water, and illuminating for 12 hours with alternating light and shade.
Test strains: staphylococcus aureus CMCC (B) 26003.
Preparation: the preparation comprises an antibacterial peptide freeze-dried preparation (I), an antibacterial peptide freeze-dried preparation (II), a freeze-dried preparation matrix (blank corresponding to the antibacterial peptide freeze-dried preparation (I)), and levofloxacin gel (0.5% preparation, Shandongming pharmaceutical industry group, Inc.).
The scheme is as follows: (1) staphylococcus aureus strains stored in liquid nitrogen were inoculated into blood plates and cultured overnight at 37 ℃.
(2) The next day, single colonies grown on blood plates were inoculated in MH liquid medium and shake-cultured for 1h at 37 ℃ in a temperature-controlled shaker.
(3) Comparing the bacteria solution to be measured with 0.5 McLeod turbidity standard solution, diluting to 5 × 10 6 CFU/ml for use.
(4) Skin modeling: shearing the back of the mouse, depilating with depilatory cream, removing hair, and polishing with 600 mesh abrasive paper until blood oozing. The subcutaneous injection concentration of the blood-seepage skin is 5 multiplied by 10 6 CFU/ml of bacterial suspension, 0.05 ml.
(5) Administration: levofloxacin gel (0.5% preparation, Shandongming pharmaceutical industry group, Inc.) 0.1ml, antibacterial peptide lyophilized preparation and lyophilized preparation matrix dosage are both 50mg, and the preparation is administered externally 1 time in the morning and evening each day for 3 consecutive days as a treatment course.
(6) After 3 days, the skin of the infected part is taken aseptically, diluted and ground to homogenate according to the volume of 1:10, 20 mul of a coating dish is taken, and viable bacteria are detected and counted.
As a result: the results of the infected skin test are shown in Table 5.
TABLE 5 Effect of lyophilized formulations of antimicrobial peptides on Staphylococcus aureus skin infections
As can be seen from Table 5, the lyophilized preparation of antibacterial peptide has a significant effect in comparison with the matrix control group and the model group, and has a better effect in resisting Staphylococcus aureus skin infection.
The antibacterial peptide freeze-dried preparation is a local external anti-infective medicament, is suitable for various infections caused by pathogenic bacteria, particularly drug-resistant bacteria, and secondary skin infections such as eczema combined infection, ulcer combined infection and the like, and has wide application prospect.
Sequence listing
<110> Jiangsu pilai pharmaceutical Biotechnology GmbH
<120> anti-infection antibacterial peptide freeze-dried preparation and preparation method thereof
<130> GNCZJ211211
<160> 4
<170> SIPOSequenceListing 1.0
<210> 1
<211> 10
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 1
Lys Trp Lys Ser Phe Leu Lys Thr Phe Lys
1 5 10
<210> 2
<211> 13
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 2
Lys Thr Val Leu His Thr Ala Leu Lys Ala Ile Ser Ser
1 5 10
<210> 3
<211> 26
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 3
Lys Trp Lys Ser Phe Leu Lys Thr Phe Lys Ser Ala Ala Lys Thr Val
1 5 10 15
Leu His Thr Ala Leu Lys Ala Ile Ser Ser
20 25
<210> 4
<211> 26
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 4
Lys Trp Lys Ser Phe Leu Lys Thr Phe Lys Ser Ala Ala Lys Thr Val
1 5 10 15
Leu His Thr Ala Leu Lys Ala Ile Ser Ser
20 25
Claims (6)
1. An antibacterial peptide freeze-dried preparation comprises the following components in parts by mass: 0.05-0.5 part of antibacterial peptide, 2-5 parts of lactose, 5-10 parts of mannitol and 0.1-1 part of sodium citrate;
Wherein the antibacterial peptide is NA L And D-NA L ；
The NA L Has the sequence shown in SEQ ID No: 3, the amino acid sequence of which is: KWKSFLKTFKSAAKTVLHTALKAISS, respectively;
the D-NA L Has the sequence shown in SEQ ID No: 4, the amino acid sequence of which is: KWKSFLKTFKSAAKTVLHTALKAISS, and all amino acids are in the D-configuration except for the L configuration of A at position 13.
2. The lyophilized formulation of antimicrobial peptide according to claim 1, wherein: the pH value of the antibacterial peptide freeze-dried preparation is 4.0-6.0.
3. The lyophilized formulation of antimicrobial peptide according to claim 1 or 2, characterized in that: the antibacterial peptide freeze-dried preparation comprises the following components in parts by mass: 0.05-0.5 part of antibacterial peptide, 3-3.5 parts of lactose, 6-6.5 parts of mannitol and 0.4-0.5 part of sodium citrate.
4. The preparation method of the lyophilized preparation of antibacterial peptide according to any one of claims 1 to 3, wherein the raw materials comprise the following substances by mass volume percent: 0.5-5 per mill of antibacterial peptide, 2-5 per mill of lactose, 5-10 percent of mannitol, 0.1-1 percent of sodium citrate and the balance of water for injection.
5. The method of claim 4, wherein: the preparation method comprises the following steps: adding the antibacterial peptide, lactose and mannitol into water for injection, stirring for dissolving, adding sodium citrate to adjust the pH value to 4.0-6.0, and supplementing the water for injection to the full amount; sterilizing and filtering with 0.22 μm filter membrane, subpackaging in penicillin bottles, and lyophilizing in vacuum lyophilizer; and after freeze-drying is finished, filling nitrogen into the box body of the vacuum freeze-drying machine, keeping the pressure of the box body to be-0.015 Mpa-0 MPa for 10-15 minutes, then pressing the plug, taking out the box and rolling the cover.
6. The production method according to claim 5, characterized in that: the lyophilization curve for the lyophilization is as follows:
a pre-freezing stage: cooling the cold trap to below-50 ℃ and maintaining for 3-4 hours;
a main sublimation stage: controlling the temperature of the heat conducting oil to rise to-10 ℃ within 1-2 hours, and maintaining for 8-12 hours; controlling the temperature of the heat conducting oil to rise to 0 ℃ within 1-2 hours, and maintaining for 15-25 hours;
and (3) analysis and drying stage: controlling the temperature of the heat conducting oil to rise to 30 ℃ within 1-2 hours, and maintaining for 5-10 hours.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110483679.4A CN113081977B (en) | 2021-04-30 | 2021-04-30 | Anti-infection antibacterial peptide freeze-dried preparation and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110483679.4A CN113081977B (en) | 2021-04-30 | 2021-04-30 | Anti-infection antibacterial peptide freeze-dried preparation and preparation method thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN113081977A CN113081977A (en) | 2021-07-09 |
| CN113081977B true CN113081977B (en) | 2022-07-29 |
Family
ID=76681396
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110483679.4A Active CN113081977B (en) | 2021-04-30 | 2021-04-30 | Anti-infection antibacterial peptide freeze-dried preparation and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113081977B (en) |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102219831A (en) * | 2011-04-18 | 2011-10-19 | 江阴普莱医药生物技术有限公司 | Antibiotic peptide as well as preparation method and application thereof |
| CN102731629A (en) * | 2012-05-21 | 2012-10-17 | 长春普莱医药生物技术有限公司 | Antibacterial peptide and application thereof |
| CN104043098A (en) * | 2014-04-04 | 2014-09-17 | 广州舒泰生物技术有限公司 | Antimicrobial peptide freeze-dried preparation for treating colpitis and preparation method thereof |
| CN105017384A (en) * | 2015-07-13 | 2015-11-04 | 长春普莱医药生物技术有限公司 | Novel antibacterial peptide and application thereof |
-
2021
- 2021-04-30 CN CN202110483679.4A patent/CN113081977B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102219831A (en) * | 2011-04-18 | 2011-10-19 | 江阴普莱医药生物技术有限公司 | Antibiotic peptide as well as preparation method and application thereof |
| CN102731629A (en) * | 2012-05-21 | 2012-10-17 | 长春普莱医药生物技术有限公司 | Antibacterial peptide and application thereof |
| CN104043098A (en) * | 2014-04-04 | 2014-09-17 | 广州舒泰生物技术有限公司 | Antimicrobial peptide freeze-dried preparation for treating colpitis and preparation method thereof |
| CN105017384A (en) * | 2015-07-13 | 2015-11-04 | 长春普莱医药生物技术有限公司 | Novel antibacterial peptide and application thereof |
Non-Patent Citations (3)
| Title |
|---|
| Rational Design of α-Helical Antimicrobial Peptides with Enhanced Activities and Specificity/Therapeutic Index*;Yuxin Chen等;《J Biol Chem》;20050401;第12316-12329页 * |
| Shiyu Sun等.Specificity and mechanism of action of alpha-helical membrane-active peptides interacting with model and biological membranes by single-molecule force spectroscopy.《Scientific Reports》.2016,第1-10页. * |
| Specificity and mechanism of action of alpha-helical membrane-active peptides interacting with model and biological membranes by single-molecule force spectroscopy;Shiyu Sun等;《Scientific Reports》;20160701;第1-10页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN113081977A (en) | 2021-07-09 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102219831B (en) | Antibiotic peptide as well as preparation method and application thereof | |
| CN105017384B (en) | A kind of antibacterial peptide and its application | |
| JP7229999B2 (en) | Daptomycin preparation | |
| EP1200463B1 (en) | Antimicrobial histone h1 compositions and methods of use thereof | |
| CN106232616A (en) | Amphipathic synthetic antibacterial peptide, its medical composition and its use | |
| CA2376894C (en) | Pharmaceutical composition comprising a protein and an ectoine | |
| CN113209273B (en) | Antibacterial peptide paint and preparation method thereof | |
| JP3100058B2 (en) | Stable pharmaceutical composition containing fibroblast growth factor | |
| CN113081977B (en) | Anti-infection antibacterial peptide freeze-dried preparation and preparation method thereof | |
| US5714458A (en) | Stable pharmaceutical compositions containing a fibroblast growth factor | |
| CN110650970B (en) | Antibacterial peptide and medicine thereof | |
| WO2020056359A1 (en) | Bacteriotherapy against proprionibacterium acnes for the treatment of acne | |
| UA125388C2 (en) | Method of preparation of lyophilized compositions | |
| CN107519136B (en) | Succinylcholine chloride freeze-dried preparation and preparation method thereof | |
| JPH04279530A (en) | Drug delivery system for administering growth factor | |
| CN112755175A (en) | Antibacterial peptide liquid composition and preparation thereof | |
| CN113134075A (en) | Antibacterial peptide emulsifiable paste and preparation method thereof | |
| US6884423B1 (en) | Antimicrobial histone H1 compositions, kits, and methods of use thereof | |
| JP2024503875A (en) | Antimicrobial peptide liquid composition and its formulation | |
| CH657989A5 (en) | BIOLOGICALLY ACTIVE SUBSTANCE, METHOD FOR PRODUCING THE SUBSTANCE, AND IMMUNOACTIVE COMPOSITION CONTAINING THE SUBSTANCE. | |
| CN113081953B (en) | Topical antibacterial peptide gel and preparation method thereof | |
| JPH0149246B2 (en) | ||
| CN111909243B (en) | Antibacterial peptide, antibacterial and itching-relieving pharmaceutical composition and application | |
| CN113717267B (en) | Antibacterial peptide pHTP-1, application thereof and product containing antibacterial peptide pHTP-1 | |
| CN116410273A (en) | Antibacterial polypeptide derived from bacillus subtilis and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |