CN113249231B - Gram-positive bacterium resisting compound derived from polar region source fungi as well as preparation method and application thereof - Google Patents

Gram-positive bacterium resisting compound derived from polar region source fungi as well as preparation method and application thereof Download PDF

Info

Publication number
CN113249231B
CN113249231B CN202110555886.6A CN202110555886A CN113249231B CN 113249231 B CN113249231 B CN 113249231B CN 202110555886 A CN202110555886 A CN 202110555886A CN 113249231 B CN113249231 B CN 113249231B
Authority
CN
China
Prior art keywords
neonectria
compound
inhibiting
growth
diseases caused
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
Application number
CN202110555886.6A
Other languages
Chinese (zh)
Other versions
CN113249231A (en
Inventor
张涛
余利岩
何文妮
毛会金
柏菁璘
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Institute of Medicinal Biotechnology of CAMS
Original Assignee
Institute of Medicinal Biotechnology of CAMS
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Institute of Medicinal Biotechnology of CAMS filed Critical Institute of Medicinal Biotechnology of CAMS
Priority to CN202110555886.6A priority Critical patent/CN113249231B/en
Publication of CN113249231A publication Critical patent/CN113249231A/en
Application granted granted Critical
Publication of CN113249231B publication Critical patent/CN113249231B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K10/00—Animal feeding-stuffs
    • A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
    • A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K20/00—Accessory food factors for animal feeding-stuffs
    • A23K20/10—Organic substances
    • A23K20/116—Heterocyclic compounds
    • A23K20/137—Heterocyclic compounds containing two hetero atoms, of which at least one is nitrogen
    • 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
    • A23L31/00—Edible extracts or preparations of fungi; Preparation or treatment thereof
    • 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
    • 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
    • A61K31/33—Heterocyclic compounds
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/06—Fungi, e.g. yeasts
    • 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
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/14—Fungi; Culture media therefor
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P17/00—Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms
    • C12P17/14—Nitrogen or oxygen as hetero atom and at least one other diverse hetero ring atom in the same ring
    • 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
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Organic Chemistry (AREA)
  • Zoology (AREA)
  • Polymers & Plastics (AREA)
  • Microbiology (AREA)
  • General Health & Medical Sciences (AREA)
  • Biotechnology (AREA)
  • Mycology (AREA)
  • Food Science & Technology (AREA)
  • Wood Science & Technology (AREA)
  • Medicinal Chemistry (AREA)
  • Biochemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Public Health (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Veterinary Medicine (AREA)
  • Animal Behavior & Ethology (AREA)
  • Genetics & Genomics (AREA)
  • Animal Husbandry (AREA)
  • Epidemiology (AREA)
  • Biomedical Technology (AREA)
  • General Engineering & Computer Science (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Botany (AREA)
  • Nutrition Science (AREA)
  • Natural Medicines & Medicinal Plants (AREA)
  • Alternative & Traditional Medicine (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Medical Informatics (AREA)
  • Physiology (AREA)
  • Molecular Biology (AREA)
  • Oncology (AREA)
  • Communicable Diseases (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Virology (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

The invention discloses a gram-positive bacterium resistant compound derived from polar region source fungi, and a preparation method and application thereof. The gram-positive bacteria resistant compound is derived from polar origin fungi, namely Neonectria sp (Neonectria sp.) CPCC401225, and the registration number of the gram-positive bacteria resistant compound in the China general microbiological culture Collection center is CGMCC No. 20277. The invention separates the enniatins compound (Enniantins) with better anti-gram-positive bacteria activity from the Neonectria fungi for the first time. The compound shown in the formula I, II or III has a simple preparation process, and is suitable for researching an anti-gram-positive bacteria lead compound or preparing an anti-gram-positive bacteria medicament.

Description

Gram-positive bacterium resisting compound derived from polar region source fungi as well as preparation method and application thereof
Technical Field
The invention relates to a gram-positive bacterium resistant compound derived from polar source fungi, a preparation method and application thereof, belonging to the field of biological medicine.
Background
Gram-positive bacteria are important pathogenic bacteria, in recent years, the increase of the infection proportion of gram-positive cocci is one of the problems that serious infectious diseases of a respiratory system are paid more attention to in clinic, and the bacteria are wide in distribution and fast in transmission and bring great difficulty to clinical treatment once infected.
In recent years, drug resistance has been increasing year by year due to abuse of antibiotics, and pulmonary infection caused by staphylococci and enterococci is more serious than other gram-positive bacteria, and endocarditis and septicemia are easily caused. Among them, methicillin-resistant staphylococcus aureus (MRSA), methicillin-resistant staphylococcus epidermidis (MRSE) and enterococcus have become extremely problematic in clinical practice. Besides, other beta-lactam antibiotics and cephalosporin antibiotics with the same structures as methicillin have drug resistance. At present, vancomycin which is commonly used clinically also gradually shows a drug resistance phenomenon. However, the newly found anti-infective drugs are few, so that the significance of finding and researching the drug which has high efficiency and low toxicity and can resist gram-positive bacteria is great.
Disclosure of Invention
The technical problem to be solved by the invention is how to prepare the drug for resisting drug-resistant gram-positive bacteria.
In order to solve the technical problems, the invention firstly provides a fungus belonging to the genus Neonectria, wherein the fungus belonging to the genus Neonectria is a fungus belonging to the genus Neonectria (Neonectria sp.) CPCC401225, and the fungus belonging to the genus Neonectria (Neonectria sp.) CPCC401225 is registered with CGMCC No.20277 in the China general microbiological culture Collection center.
The invention also provides a culture of the Neonectria fungus, wherein the culture is obtained by culturing the Neonectria fungus (Neonectria sp.) CPCC401225 in a microbial culture medium (namely a fermentation product, such as a fermentation liquid containing the Neonectria fungus and a substance secreted into a liquid culture medium, or a solid fermentation product containing the Neonectria fungus and a substance secreted into a solid culture medium).
The invention also provides a microbial inoculum, which contains the Neonectria fungus or/and metabolites of the Neonectria fungus or/and the culture.
The microbial inoculum can have at least one of the following functions:
a1, treating and/or preventing diseases caused by staphylococcus aureus,
a2, treating and/or preventing diseases caused by enterococcus faecium,
a3, treatment and/or prevention of diseases caused by Escherichia coli,
a4, treating and/or preventing diseases caused by Klebsiella pneumoniae,
a5, treating and/or preventing diseases caused by Acinetobacter baumannii,
a6, inhibiting the growth of staphylococcus aureus,
a7, inhibiting the growth of enterococcus faecium,
a8, inhibiting the growth of Escherichia coli,
a9, inhibiting the growth of Klebsiella pneumoniae,
a10, inhibiting the growth of acinetobacter baumannii.
Above, the metabolite may be a fermentation broth of the fungus of the genus Neonectria. The fermentation broth of the fungus belonging to the genus Neonectria can be prepared as follows: culturing the Neonectria fungus in a liquid fermentation medium, and collecting fermentation liquor (containing the Neonectria fungus and substances secreted into the liquid medium), wherein the fermentation liquor is a metabolite of the Neonectria fungus.
The active ingredients of the microbial inoculum can be the Neonectria fungus or/and metabolites of the Neonectria fungus or/and cultures of the Neonectria fungus, and can also contain other biological ingredients or non-biological ingredients, and the other active ingredients of the microbial inoculum can be determined by the technicians in the field according to the effects of the microbial inoculum.
In the above microbial inoculum, the microbial inoculum may further comprise a carrier. The carrier may be a solid carrier or a liquid carrier.
In the microbial inoculum, the dosage form of the microbial inoculum can be various dosage forms, including but not limited to liquid, emulsion, suspending agent, powder, granules, wettable powder or water dispersible granules and the like.
The invention also provides a product containing the Neonectria fungus, the culture or the microbial inoculum. Such products include, but are not limited to, pharmaceuticals, foods, nutraceuticals, animal feeds, or animal feed additives.
In the above medicament, the medicament may comprise a pharmaceutically acceptable carrier. The carrier material includes, but is not limited to, water-soluble carrier materials (such as polyethylene glycol, polyvinylpyrrolidone, organic acids, etc.), poorly soluble carrier materials (such as ethyl cellulose, cholesterol stearate, etc.), enteric carrier materials (such as cellulose acetate phthalate, carboxymethyl cellulose, etc.). Among these, water-soluble carrier materials are preferred. The materials can be prepared into various dosage forms, including but not limited to tablets, capsules, dripping pills, aerosols, pills, powders, solutions, suspensions, emulsions, granules, liposomes, microspheres, transdermal agents, buccal tablets, suppositories, freeze-dried powder injections and the like. Can be common preparation, sustained release preparation, controlled release preparation and various microparticle drug delivery systems. In order to prepare the unit dosage form into tablets, various carriers well known in the art can be widely used. Examples of the carrier are, for example, diluents and absorbents such as starch, dextrin, calcium sulfate, lactose, mannitol, sucrose, sodium chloride, glucose, urea, calcium carbonate, kaolin, microcrystalline cellulose, aluminum silicate and the like; wetting agents and binders such as water, glycerin, polyethylene glycol, ethanol, propanol, starch slurry, dextrin, syrup, honey, glucose solution, acacia slurry, gelatin slurry, sodium carboxymethylcellulose, shellac, methyl cellulose, potassium phosphate, polyvinylpyrrolidone and the like; disintegrating agents such as dried starch, alginate, agar powder, brown algae starch, sodium bicarbonate and citric acid, calcium carbonate, polyoxyethylene, sorbitol fatty acid ester, sodium dodecylsulfate, methyl cellulose, ethyl cellulose, etc.; disintegration inhibitors such as sucrose, glyceryl tristearate, cacao butter, hydrogenated oil and the like; absorption accelerators such as quaternary ammonium salts, sodium lauryl sulfate and the like; lubricants, for example, talc, silica, corn starch, stearate, boric acid, liquid paraffin, polyethylene glycol, and the like. The tablets may be further formulated into coated tablets, such as sugar-coated tablets, film-coated tablets, enteric-coated tablets, or double-layer and multi-layer tablets. In order to prepare the dosage form for unit administration into a pill, various carriers well known in the art can be widely used. Examples of the carrier are, for example, diluents and absorbents such as glucose, lactose, starch, cacao butter, hydrogenated vegetable oil, polyvinylpyrrolidone, kaolin, talc and the like; binders such as acacia, tragacanth, gelatin, ethanol, honey, liquid sugar, rice paste or batter, etc.; disintegrating agents, such as agar powder, dried starch, alginate, sodium dodecylsulfate, methylcellulose, ethylcellulose, etc. In order to prepare the unit dosage form into suppositories, various carriers known in the art can be widely used. As examples of the carrier, there may be mentioned, for example, polyethylene glycol, lecithin, cacao butter, higher alcohols, esters of higher alcohols, gelatin, semisynthetic glycerides and the like. In addition, colorants, preservatives, flavors, flavorings, sweeteners or other materials may also be added to the pharmaceutical preparation, if desired.
The above medicine can be human medicine or animal medicine (veterinary medicine). In the above medicines, the dosage forms of the medicines include, but are not limited to, tablets, capsules, dropping pills, aerosols, pills, powders, solutions, suspensions, emulsions, granules, liposomes, microspheres, transdermal agents, buccal tablets, suppositories or freeze-dried powder injections and the like.
The food can be solid food, or liquid food such as beverage.
The product may have at least one of the functions of A1-A16:
a1, treating and/or preventing diseases caused by staphylococcus aureus,
a2, treating and/or preventing diseases caused by enterococcus faecium,
a3, treatment and/or prevention of diseases caused by Escherichia coli,
a4, treating and/or preventing diseases caused by Klebsiella pneumoniae,
a5, treating and/or preventing diseases caused by Acinetobacter baumannii,
a6, inhibiting the growth of staphylococcus aureus,
a7, inhibiting the growth of enterococcus faecium,
a8, inhibiting the growth of Escherichia coli,
a9, inhibiting the growth of Klebsiella pneumoniae,
a10, inhibiting the growth of Acinetobacter baumannii.
The application of the Neonectria fungus, the culture or the microbial inoculum in preparing any functional product also belongs to the protection scope of the invention:
a1, treating and/or preventing diseases caused by staphylococcus aureus,
a2, treating and/or preventing diseases caused by enterococcus faecium,
a3, treatment and/or prevention of diseases caused by Escherichia coli,
a4, treating and/or preventing diseases caused by Klebsiella pneumoniae,
a5, treating and/or preventing diseases caused by Acinetobacter baumannii,
a6, inhibiting the growth of staphylococcus aureus,
a7, inhibiting the growth of enterococcus faecium,
a8, inhibiting the growth of Escherichia coli,
a9, inhibiting the growth of Klebsiella pneumoniae,
a10, inhibiting the growth of acinetobacter baumannii.
The application of the Neonectria fungus, the culture or the microbial inoculum in preparing the compound 1, the compound 2 and/or the compound 3 also belongs to the protection scope of the invention;
the compound 1 is shown as a formula I, the compound 2 is shown as a formula II, and the compound 3 is shown as a formula III:
Figure GDA0003647622340000041
Figure GDA0003647622340000051
the present invention also provides a process for the preparation of a compound, the process comprising: culturing a fungus of the genus Neonectria in a culture medium to obtain a fermentation product, and obtaining the compound from the fermentation product; the Neonectria fungus is Neonectria fungus (Neonectria sp.) CPCC401225, which is registered with CGMCC No.20277 in China general microbiological culture Collection center; the structural formula of the compound is shown as formula I or II or III:
Figure GDA0003647622340000052
in the above method, the culture medium is a culture medium for a fungus of the genus Neonectria (a culture medium capable of culturing a fungus of the genus Neonectria), and may be a solid culture medium, a semi-solid culture medium, or a liquid culture medium. The microbial culture medium may be a rice culture medium, a potato dextrose agar medium, or other fungal fermentation media well known to those skilled in the art, and the like. The rice culture medium can be made of rice and water, can also be made of rice, water and inorganic salt, can also be made of rice, water and nitrogen source, and can also be made of rice, water, nitrogen source and inorganic salt. The potato glucose agar medium can be prepared from potato, glucose, agar and water, or from potato, glucose, agar, water and inorganic salt, or from potato, carbon source, agar, water and nitrogen source, or from potato, carbon source, agar, nitrogen source and inorganic salt. The other fungal fermentation media known to those skilled in the art may be prepared from one or more of a fast-acting, slow-acting carbon source, one or more of a fast-acting, slow-acting nitrogen source, water and/or inorganic salts, and the like.
In the above method, the rice may be rice or brown rice. The rice or brown rice is a product of rice. The rice refers to a fruit from which rice husk is not removed, and is composed of husk, pericarp, seed coat, endosperm, aleurone layer, endosperm and embryo. The brown rice is the product of rice with husk removed and other parts reserved; rice refers to a product in which only the endosperm is retained and the rest of the rice is completely removed. The carbon source is a nutrient for microbial growth, and is a carbon-containing compound, including quick-acting and slow-acting carbon sources such as saccharides, grease, organic acids, organic acid esters and micromolecular alcohols. The nitrogen source is a substance for providing nitrogen elements required by the nutrition of microorganisms, and comprises quick-acting and slow-acting nitrogen sources such as peanut cake powder, soybean cake powder, yeast powder, peptone, ammonia water, ammonium salt, nitrate and the like.
In the above method, the temperature of the culture may be 15 ℃. The culture time may be 30-40 days.
In the above preparation method, obtaining the compound from the fermentation product comprises the steps of: 1) extracting the fermentation product with an organic solvent taking ethyl acetate as a leaching solution, and collecting a leaching product containing the compound; 2) subjecting the extract to adsorption chromatography using silica gel as an adsorbent, eluting with a solution capable of dissolving the compound, and collecting an eluate containing the compound; 3) separating the compound from the eluate.
In the above preparation method, in the 2), the type of the silica gel in the adsorption chromatography may be chromatography silica gel H. The solution in which the compound is soluble may be a dichloromethane methanol solution. The elution procedure used in the silica gel column chromatography was as follows: 1) eluting 3 column volumes with dichloromethane; 2) then, using 200: 1 (a liquid consisting of dichloromethane and methanol in a volume ratio of dichloromethane to methanol of 200: 1) eluting 3 column volumes; 3) then, using 200: 3 (a liquid consisting of dichloromethane and methanol in a volume ratio of dichloromethane to methanol of 200: 3) elution for 3 column volumes; 4) and then 200: 5 dichloromethane-methanol solution (a liquid consisting of dichloromethane and methanol in a volume ratio of dichloromethane to methanol of 200: 5) eluting 3 column volumes; 5) and then 200: 7 (a liquid consisting of dichloromethane and methanol in a volume ratio of dichloromethane to methanol of 200: 7) eluting 3 column volumes; 6) and then 200: 9 (a liquid consisting of dichloromethane and methanol in a volume ratio of dichloromethane to methanol of 200: 9) elution for 3 column volumes; the eluted liquid was collected continuously from the start of the elution procedure, and the 16 th column volume to 18 th column volume (i.e., 200: 9 methylene chloride-methanol eluted liquid) was collected and designated fraction E. The fraction E contains the compound 1, the compound 2 and the compound 3.
In the above process, 3) washing from the washingThe separation in the liquid removal to obtain the compound can be realized by adopting medium-pressure ODS liquid-phase preparation and separation. The fraction E was subjected to liquid-phase preparation of a separation medium using medium-pressure ODS, and the separation medium was Lichroprep RP-C18. The elution procedure used in the medium-pressure ODS liquid-phase preparative separation was as follows: performing linear gradient elution with methanol-water solution (the methanol-water solution is a liquid composed of methanol and water, the volume concentration of methanol is linearly increased from 10% to 100% within 180 min), the flow rate is 15mL/min, and collecting t R The (retention time) was the peak eluted in 145-158min, which was designated as fraction Fr.E8. E8 contains said compound 1, said compound 2 and said compound 3.
The 3) above may further comprise subjecting the fraction fr.e8 to column chromatography with gel LH20 to obtain the compound 1, the compound 2 and the compound 3. The elution procedure used in gel LH20 column chromatography was as follows: the method comprises the following steps of 1: 1 dichloromethane-methanol solution (liquid consisting of dichloromethane and methanol in a 1: 1 ratio by volume of dichloromethane to methanol) eluted for 2 column volumes, 565.2mL column volumes. The eluted liquid is collected continuously from the beginning of the elution program, 20mL of the eluted liquid is collected in each fraction, and the collected eluted liquid of the 7 th fraction to the 8 th fraction is named as Fr.E8-4.
The step 3) can further comprise the step of separating the Fr.E8-4 by using SB-C3 liquid phase preparation to obtain a compound 1, a compound 2 and a compound 3. The separation chromatographic column used in the liquid phase preparation of the SB-C3 is Agilent SB-C3. The mobile phase is 65% acetonitrile-water (liquid consisting of acetonitrile and water, the volume ratio of the acetonitrile to the water in the liquid is 65: 35), the flow rate is 2mL/min, and an elution peak with the retention time of 22.5min is collected to obtain a compound 1; collecting the elution peak with retention time of 26.5min to obtain a compound 2; the elution peak with retention time of 28.5min was collected to give compound 3.
The invention also provides a bacterial inhibitor or antibacterial medicament comprising a compound of formula I or II or III:
Figure GDA0003647622340000071
Figure GDA0003647622340000081
the term "pharmaceutically acceptable salt" refers to salts which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals without excessive toxicity, irritation, allergic response, and the like, commensurate with a reasonable benefit/risk ratio. Pharmaceutically acceptable salts are well known in the art. Pharmaceutically acceptable salts are described in detail, for example, in s.m. berge, et al, j.pharmaceutical Sciences,1977,66: 1. The salts may be prepared by reacting the free base functionality of the compounds of the present invention with a suitable organic acid.
The active ingredient of the bacterial inhibitor or antibacterial drug may be the compound or a pharmaceutically acceptable salt thereof, and the active ingredient of the bacterial inhibitor or antibacterial drug may further contain other biological or non-biological components, and other active ingredients may be determined by those skilled in the art according to antiviral effects.
The bacteria may be staphylococcus aureus, enterococcus faecium, escherichia coli, klebsiella pneumoniae, or acinetobacter baumannii.
The invention also provides methods of inhibiting gram-positive bacteria from infecting an animal.
The invention provides a method for inhibiting gram-positive bacteria from infecting animals, which comprises administering a compound shown as a formula I or a pharmaceutically acceptable salt thereof to a receptor animal to inhibit gram-positive bacteria from infecting animals.
The invention also provides a method for treating or/and preventing influenza.
The method for treating or/and preventing influenza provided by the invention comprises the step of administering the compound shown as the formula I, II or III or the pharmaceutically acceptable salt thereof to a receptor animal to treat or/and prevent the influenza.
As noted above, the gram-positive bacteria can be embodied to inhibit replication of gram-positive bacteria.
As above, the gram-positive bacterium may be a gram-positive influenza bacterium. The gram-positive influenza bacteria can be gram-positive bacteria. The gram-positive bacteria resistant medicine can be a medicine for treating or/and preventing influenza.
In the present invention, the animal may be a mammal, such as a human; the animal can also be other animals, such as birds, infected with gram-positive bacteria other than mammals.
As mentioned above, the bacterial inhibitor or antibacterial agent may further comprise a suitable carrier or excipient in addition to the compound represented by formula I, II or III or a pharmaceutically acceptable salt thereof. The carrier material herein includes, but is not limited to, water-soluble carrier materials (e.g., polyethylene glycol, polyvinylpyrrolidone, organic acids, etc.), poorly soluble carrier materials (e.g., ethyl cellulose, cholesterol stearate, etc.), enteric carrier materials (e.g., cellulose acetate phthalate, carboxymethyl cellulose, etc.). Among these, water-soluble carrier materials are preferred. The materials can be prepared into various dosage forms, including but not limited to tablets, capsules, dripping pills, aerosols, pills, powders, solutions, suspensions, emulsions, granules, liposomes, transdermal agents, buccal tablets, suppositories, freeze-dried powder injections and the like. Can be common preparation, sustained release preparation, controlled release preparation and various microparticle drug delivery systems. In order to prepare the unit dosage form into tablets, various carriers well known in the art can be widely used. Examples of the carrier are, for example, diluents and absorbents such as starch, dextrin, calcium sulfate, lactose, mannitol, sucrose, sodium chloride, glucose, urea, calcium carbonate, kaolin, microcrystalline cellulose, aluminum silicate and the like; wetting agents and binders such as water, glycerin, polyethylene glycol, ethanol, propanol, starch slurry, dextrin, syrup, honey, glucose solution, acacia slurry, gelatin slurry, sodium carboxymethylcellulose, shellac, methyl cellulose, potassium phosphate, polyvinylpyrrolidone and the like; disintegrating agents such as dried starch, alginate, agar powder, brown algae starch, sodium bicarbonate and citric acid, calcium carbonate, polyoxyethylene, sorbitol fatty acid ester, sodium dodecylsulfate, methyl cellulose, ethyl cellulose, etc.; disintegration inhibitors such as sucrose, glyceryl tristearate, cacao butter, hydrogenated oil and the like; absorption accelerators such as quaternary ammonium salts, sodium lauryl sulfate and the like; lubricants, for example, talc, silica, corn starch, stearate, boric acid, liquid paraffin, polyethylene glycol, and the like. The tablets may be further formulated into coated tablets, such as sugar-coated tablets, film-coated tablets, enteric-coated tablets, or double-layer and multi-layer tablets. In order to prepare the dosage form for unit administration into a pill, various carriers well known in the art can be widely used. Examples of the carrier are, for example, diluents and absorbents such as glucose, lactose, starch, cacao butter, hydrogenated vegetable oil, polyvinylpyrrolidone, kaolin, talc and the like; binders such as acacia, tragacanth, gelatin, ethanol, honey, liquid sugar, rice paste or batter, etc.; disintegrating agents, such as agar powder, dried starch, alginate, sodium dodecylsulfate, methylcellulose, ethylcellulose, etc. In order to prepare the unit dosage form as a suppository, various carriers well known in the art may be widely used. As examples of the carrier, there may be mentioned, for example, polyethylene glycol, lecithin, cacao butter, higher alcohols, esters of higher alcohols, gelatin, semisynthetic glycerides and the like. In order to prepare the unit dosage form into preparations for injection, such as solutions, emulsions, lyophilized powders and suspensions, all diluents commonly used in the art, for example, water, ethanol, polyethylene glycol, 1, 3-propanediol, ethoxylated isostearyl alcohol, polyoxylated isostearyl alcohol, polyoxyethylene sorbitol fatty acid esters, etc., can be used. In addition, for the preparation of isotonic injection, sodium chloride, glucose or glycerol may be added in an appropriate amount to the preparation for injection, and conventional cosolvents, buffers, pH adjusters and the like may also be added. In addition, colorants, preservatives, flavors, flavorings, sweeteners or other materials may also be added to the pharmaceutical preparation, if desired. The preparation can be used for injection administration, including subcutaneous injection, intravenous injection, intramuscular injection, intracavity injection and the like; for administration to the cavities, e.g., rectally and vaginally; administration to the respiratory tract, e.g., nasally; administration to the mucosa. The above route of administration is preferably by injection.
As indicated above, the compounds of formula I include racemates, enantiomers, diastereomers, tautomers, polymorphs, pseudopolymorphs, amorphous forms, hydrates or solvates.
The compound shown in the formula I, II or III is derived from Neonectria fungi, and the enniatins compound (Enniantins) with better anti-gram-positive bacteria activity is obtained by separating from the Neonectria fungi for the first time. The compound shown in the formula I, II or III has a simple preparation process, and is suitable for researching an anti-gram-positive bacteria lead compound or preparing an anti-gram-positive bacteria medicament.
Deposit description
And (3) classification and naming: neonectria sp.
The strain number is as follows: CPCC401225
The name of the depository: china general microbiological culture Collection center
The preservation unit is abbreviated as: CGMCC (China general microbiological culture Collection center)
The address of the depository: west road No. 1, north chen, chaoyang district, beijing, zip code: 100101
The preservation date is as follows: 10 and 12 months in 2020
Registration number of the preservation center: CGMCC No.20277
Drawings
FIG. 1 is a UV spectrum of Compound 1.
FIG. 2 is a UV spectrum of Compound 2.
FIG. 3 is a UV spectrum of Compound 3.
FIG. 4 is a mass spectrum of Compound 1.
FIG. 5 is a mass spectrum of Compound 2.
FIG. 6 is a mass spectrum of Compound 3.
FIG. 7 shows that Compound 1 is dissolved in DMSO-d 6 In (1) 1 H-NMR spectrum.
FIG. 8 shows that Compound 2 is dissolved in DMSO-d 6 In (1) 1 H-NMR spectrum.
FIG. 9 shows that Compound 3 is dissolved in DMSO-d 6 In (1) 1 H-NMR spectrum.
FIG. 10 shows that Compound 1 is dissolved in DMSO-d 6 In (1) 13 C-NMR spectrum.
FIG. 11 shows that Compound 2 is dissolved in DMSO-d 6 In (1) 13 C-NMR spectrum.
FIG. 12 shows that Compound 3 is dissolved in DMSO-d 6 In (1) 13 C-NMR spectrum.
Detailed Description
The present invention is described in further detail below with reference to specific embodiments, which are given for the purpose of illustration only and are not intended to limit the scope of the invention. The examples provided below serve as a guide for further modifications by a person skilled in the art and do not constitute a limitation of the invention in any way.
The experimental procedures in the following examples, unless otherwise indicated, are conventional and are carried out according to the techniques or conditions described in the literature in the field or according to the instructions of the products. Materials, reagents, instruments and the like used in the following examples are commercially available unless otherwise specified. The quantitative tests in the following examples, all set up three replicates and the results averaged.
Example 1 preparation of gram-Positive bacteria-resisting Compound and gram-Positive bacteria-resisting Activity thereof
Method for preparing anti-gram-positive bacteria compound by fermentation
1. Isolation and characterization of Neonectria fungus (Neonectria sp.) CPCC401225
1.1 fungi of the genus Neonectria (Neonectria sp.) CPCC401225 was isolated from a sample isolated from the lichen arctica (Sanionia unicatata (Hedw.) Loeske). The method comprises the following steps: collected Arctic moss is put into a self-sealing bag and taken back to a laboratory for treatment. Weighing 1g of young root, rinsing with 75% ethanol water solution for 30s, washing with sterile water for 3 times, cutting the young root with sterile scissors, placing into a sterilized mortar, and adding quartz sand for grinding. Adding the grinding fluid into a centrifuge tube filled with 9mL of sterile water, wherein the sample dilution is 10 -1 After shaking up, 10 are prepared in sequence -2 、10 -3 、10 -4 、10 -5 And (4) diluting the solution in a gradient manner. The different gradient dilutions were pipetted at 200. mu.L each and applied evenly to PDA plates, each concentration was repeated 3 times. Culturing in 15 deg.C incubator. 5-7dThen, according to the color, edge, shape, transparency, etc. of the colony, selecting single colony, streaking 3 times on PDA culture medium plate, culturing, and storing in refrigerator after growing. The strain numbered CPCC401225 was taken for the following identification.
The preservation method of the strain CPCC401225 comprises the following steps: storing in 20% glycerol freezing tube at-80 deg.C.
1.2 identification of fungi of the genus Neonectria (Neonectria sp.) CPCC401225
1.2.1 morphological characteristics
The bacterial colony of the strain CPCC401225 is loose in texture, dry in appearance, opaque, light yellow brown, developed in aerial hyphae, and in the shape of a tight or loose cobweb, villous or cotton flocculent; the colony is tightly connected with the culture medium.
1.2.2 molecular biological identification
Selecting a bacterial strain CPCC401225 from a solid culture medium, putting the bacterial strain CPCC401225 in a sterile 1.5mL centrifuge tube, adding liquid nitrogen, cooling, and rapidly grinding a sample into powder by using a special glass grinding rod; adding 300 μ L of 2 × CTAB extract, mixing, and water bath at 95 deg.C for 2 min; adding phenol-chloroform mixture (phenol: chloroform: isoamyl alcohol (25: 24: 1, V/V/V)) at an equal volume (300. mu.L), shaking, mixing, and centrifuging at 12000rpm for 5 min; transferring the supernatant into a new 1.5mL sterile centrifuge tube, adding isopropanol with the same volume, shaking, mixing uniformly, transferring into a silica membrane adsorption centrifuge column, standing for 2min, and centrifuging at 12000rpm for 30 s; discarding the liquid, adding 500 μ L EB washing solution into the silica membrane adsorption centrifugal column, and centrifuging at 12000rpm for 30 s; centrifugally drying the silicon column at 12000rpm for 2 min; transferring the silica membrane adsorption centrifugal column into a new 1.5mL sterile centrifuge tube, adding 50 mu L TE eluent at 65 ℃, and centrifuging at 12000rpm for 30 s; discard the silica column, collect 50 μ L liquid in the centrifuge tube to obtain the crude extract of total DNA. The crude extract of the DNA was subjected to sequencing by Shanghai Biotech Co. The sequence determination results are submitted to NCBI and compared with the corresponding gene sequences of the effectively described strains in related genera, and the affiliated groups of the strains are preliminarily determined.
The result shows that the ITS gene sequence of the strain CPCC401225 is shown as the sequence 1 in the sequence table, and the ITS gene sequence has 99.44 percent of similarity with the ITS gene sequence of the strain Neonectria ramularialian strain CBS 182.36.
Strain CPCC401225 was determined to be a fungus of the genus Neonectria (Neonectria sp) based on the morphological characteristics of strain CPCC401225 and ITS gene sequences.
The strain CPCC401225 has been preserved in China general microbiological culture Collection center (CGMCC) at 10 months and 12 days in 2020, with the preservation number of CGMCC No. 20277. Hereinafter referred to as Neonectria fungus (Neonectria sp.) CPCC 401225.
2. Fermentation culture
(1) A strain of CPCC401225, spores of a fungus belonging to the genus Neonectria (Neonectria sp.), was inoculated into a slant culture medium and cultured at 15 ℃ for 7 days to obtain a slant strain.
Wherein the slant culture medium comprises potato extract powder, glucose, agar and water. The concentrations of the components in the slant culture medium are respectively as follows: 4g/L of potato extract powder, 20g/L of glucose and 15g/L of agar; the pH value of the slant culture medium is 5.6 +/-0.2.
(2) 100mL of seed medium was placed in 500mL glass bottles, sterilized at 121 ℃ for 20 minutes, and inoculated with mycelia taken from the slant medium on which the culture was completed in step (1). The culture was carried out at 15 ℃ for 7 days with shaking (200rpm) to obtain a seed solution.
Wherein the seed culture medium comprises potato extract powder, glucose and water. The concentrations of the components in the seed culture medium are respectively as follows: soaking potato powder in a concentration of 4 g/L; 20g/L of glucose; the pH value of the liquid culture medium is 5.6 +/-0.2.
(3) A500 mL Erlenmeyer flask containing 100g of rice was soaked in 100mL of water and sterilized at 121 ℃ for 20 minutes to obtain a fermentation medium. And (3) inoculating the seed solution obtained in the step (2) into a fermentation culture medium (10 mL of the seed solution is inoculated into each bottle of the fermentation culture medium), and statically culturing at 15 ℃ for 40 days to obtain a fermentation product (after 7 days of culture, the culture medium is scattered to small pieces in an aseptic environment, and the culture is continued, wherein the culture period can be carried out by shaking irregularly so as to ensure the air to enter in the culture process).
Secondly, separating and purifying the compound resisting gram-positive bacteria and identifying
1. Separation and purification of compounds resisting gram-positive bacteria
1.1 fermentation product extraction
Ultrasonically extracting the fermentation product obtained in the first step with ethyl acetate for 3 times, wherein the extraction is 5L multiplied by 3, filtering the obtained ethyl acetate solution by using a Buchner funnel, and concentrating the filtrate under reduced pressure to obtain a crude ethyl acetate extract.
1.2 preparation of Compound 1, Compound 2 and Compound 3
The crude ethyl acetate extract obtained in step 1.1 is separated by chromatography silica gel H, the column volume is 6358.5mL, and the silica gel H is a product of Qingdao ocean chemical industry Co. The elution procedure used in the gel column chromatography was as follows: 1) eluting 3 column volumes with dichloromethane; 2) then, using 200: 1 (a liquid consisting of dichloromethane and methanol in a volume ratio of dichloromethane to methanol of 200: 1) eluting 3 column volumes; 3) then, using 200: 3 (a liquid consisting of dichloromethane and methanol in a volume ratio of dichloromethane to methanol of 200: 3) elution for 3 column volumes; 4) and then 200: 5 dichloromethane-methanol solution (a liquid consisting of dichloromethane and methanol in a volume ratio of dichloromethane to methanol of 200: 5) eluting 3 column volumes; 5) and then 200: 7 (a liquid consisting of dichloromethane and methanol in a volume ratio of dichloromethane to methanol of 200: 7) eluting 3 column volumes; 6) and then 200: 9 (a liquid consisting of dichloromethane and methanol in a volume ratio of dichloromethane to methanol of 200: 9) elution for 3 column volumes; the eluted liquid was collected continuously from the start of the elution procedure, and the 16 th column volume to 18 th column volume (i.e., 200: 9 methylene chloride-methanol eluted liquid) was collected and designated fraction E.
Fraction E was subjected to preparative separation of the compound using a medium pressure ODS liquid phase using a medium of Lichroprep RP-C18 (product of Angela technology Co., Ltd.), 30 mm. times.200 mm (diameter. times.length), a column volume of 141.3mL, and a column pressure of 20 MPa. The elution procedure used in the medium-pressure ODS liquid-phase preparative separation was as follows: performing linear gradient elution with methanol water solution (the methanol water solution is a liquid composed of methanol and water, and the volume concentration of methanol is 1)Linear from 10% to 100% in 80 min) at a flow rate of 15mL/min, and collecting t R The (retention time) was the peak eluted in 145-158min, which was designated as fraction Fr.E8.
E8 fraction Fr was separated by column chromatography on gel LH20, 20mm X1800 mm (diameter. times. length) in a column volume of 565.2mL, using gel LH20 from GE. The elution procedure used in gel LH20 column chromatography was as follows: the method comprises the following steps of 1: 1 dichloromethane-methanol solution (liquid consisting of dichloromethane and methanol in a volume ratio of 1: 1) eluted 2 column volumes. The eluted liquid is collected continuously from the beginning of the elution program, 20mL of the eluted liquid is collected in each fraction, and the collected eluted liquid of the 7 th fraction to the 8 th fraction is named as Fr.E8-4.
Fr.E8-4 was subjected to liquid phase preparative separation using SB-C3 to give Compound 1, Compound 2 and Compound 3 using Agilent SB-C3, 9.6 mm. times.250 mm (diameter. times.length) column volume 72.34 mL. The mobile phase is 65% acetonitrile-water (liquid consisting of acetonitrile and water, the volume ratio of the acetonitrile to the water in the liquid is 65: 35), the flow rate is 2mL/min, and an elution peak with the retention time of 22.5min is collected to obtain a compound 1; collecting the elution peak with retention time of 26.5min to obtain a compound 2; the elution peak with retention time of 28.5min was collected to give compound 3.
2. Identification of gram-positive bacteria-resistant compounds 1-3
Identifying the obtained compounds 1-3:
(1) appearance: the compounds 1 to 3 are white amorphous powders.
(2) Solubility: the compounds 1 to 3 are easily soluble in methanol, acetonitrile, DMSO, etc.
(3) Ultraviolet spectrum identification: FIG. 1 shows that the ultraviolet spectrum of an acetonitrile solution of Compound 1 has a maximum absorption peak at 205 nm. FIG. 2 shows that the UV spectrum of the acetonitrile solution of Compound 2 has a maximum absorption peak at 210 nm. FIG. 3 shows that the UV spectrum of an acetonitrile solution of Compound 3 has a maximum absorption peak at 195 nm.
(4) Mass spectrum: FIG. 4 is an ESIMS mass spectrum of Compound 1, showing that [ M + Na ] thereof] + The peak was m/z 662.42. FIG. 5 is an ESIMS mass spectrum of Compound 2, shown[M+Na] + Peak M/z 676.25, ESIMS mass spectrum of Compound 3 showing [ M + Na ] in FIG. 6] + The peak was m/z 690.58, and ESIMS testing was performed using a Thermo Scientific LTQ Orbitrap XL mass spectrometry system. Methanol is used as a solvent.
(5) Nuclear magnetic resonance spectroscopy: FIG. 7, FIG. 8 and FIG. 9 are for Compound 1, Compound 2 and Compound 3, respectively 1 H-NMR spectrum. FIG. 10, FIG. 11 and FIG. 12 are for Compound 1, Compound 2 and Compound 3, respectively 13 C-NMR spectra, compared with the literature, were studied for the NMR spectra of the three compounds and assigned to the NMR signals, see Table 1.
NMR measurements of Compound 1, Compound 2 and Compound 3 Using a Bruker 600MHz instrument (R: (R)) 1 H 600MHz； 13 C150 MHz), the solvent is DMSO-d 6 (solvent Peak correction. delta H 2.50/δ C 39.5)。
TABLE 1 assignment of peaks in NMR spectra of Compound 1, Compound 2 and Compound 3: ( 1 H NMR 600MHz； 13 C NMR 150MHz；DMSO-d 6 )
Figure GDA0003647622340000141
Figure GDA0003647622340000151
The structures of compound 1 (compound shown in formula I), compound 2 (compound shown in formula II) and compound 3 (compound shown in formula III) are finally determined as follows:
Figure GDA0003647622340000152
3. detection of gram-positive bacterium-resistant Activity of Compounds 1 to 3
MIC determination experiments were performed using the microdilution method. The test bacteria include Staphylococcus aureus (ATCC 33591, Staphylococcus aureus subsp. aureus Rosenbach, methicillin-resistant strain), Enterococcus faecium (ATCC700221, Enterococcus faecium (Orla-Jensen) Schleifer and Kilpper-Balz, vancomycin-resistant strain), Escherichia coli (ATCC 25922, Escherichia coli (Migula) Castellani and Chalmers), Klebsiella pneumoniae (ATCC 700603, Escherichia pneoniae (Schroeter) Trevisan), and Acinetobacter baumannii (ATCC 19606, Bacillus anitratum Schaub and Hauber).
Bacterial liquid culture: the frozen strains or single colonies picked from the plates were inoculated in 1Ml MH broth for about 18 hours overnight.
Sample dilution: stock solutions of compounds 1-3 were diluted to 256. mu.g/ml with MH broth, 100. mu.l of this concentration sample solution was pipetted into the first row of a 96-well plate, and 100. mu.l of MH broth was pipetted into each of the remaining rows. Sucking 100 μ l MH broth with a row gun, adding into the first row, mixing, sucking 100 μ l of the above mixed solution, adding into the second row, and diluting twice to the last row, wherein the concentration of the sample in each row of each well is 128, 64, 32, 16, 8, 4, 2, 1, 0.5, 0.25, 0.125, 0.0625, and 0.03125 μ g/ml. Levofloxacin (levofloxacin) was used as a control.
Inoculating experimental bacteria: taking the concentration as 10 6 CFU/ml of the bacterial suspension was added to each well at 10. mu.l per well, and the final concentration of bacteria in each well was 5X 10 6 And (3) CFU/ml, culturing at the constant temperature of 37 ℃ for 18h, and observing the result, wherein the minimum concentration of the medicine contained in the bacteria-free growing hole is the Minimum Inhibitory Concentration (MIC).
The results are shown in Table 2, and show that the compounds 1-3 have good inhibition effects on staphylococcus aureus and enterococcus faecium, and the MIC values of the compound 1 on the staphylococcus aureus and the enterococcus faecium are 32 mu g/ml and 8 mu g/ml respectively; MIC values of the compound 2 to staphylococcus aureus and enterococcus faecium are respectively 32 mu g/ml and 8 mu g/ml, MIC values of the compound 3 to staphylococcus aureus and enterococcus faecium are respectively 64 mu g/ml and 32 mu g/ml, and the compounds 1-3 can also inhibit escherichia coli, klebsiella pneumoniae and acinetobacter baumannii to a certain extent.
TABLE 2 MIC test results for antibacterial Activity of Compound 1, Compound 2 and Compound 3
Figure GDA0003647622340000161
The present invention has been described in detail above. It will be apparent to those skilled in the art that the invention can be practiced in a wide range of equivalent parameters, concentrations, and conditions without departing from the spirit and scope of the invention and without undue experimentation. While the invention has been described with reference to specific embodiments, it will be appreciated that the invention can be further modified. In general, this application is intended to cover any variations, uses, or adaptations of the invention following, in general, the principles of the invention and including such departures from the present disclosure as come within known or customary practice within the art to which the invention pertains. The use of some of the essential features is possible within the scope of the claims attached below.
<110> institute of medical and Biotechnology of Chinese academy of medical sciences
<120> gram-positive bacterium resisting compound derived from polar region source fungi, and preparation method and application thereof
<160> 1
<170> PatentIn version 3.5
<210> 1
<211> 488
<212> DNA
<213> Neonectria sp.
<400> 1
ggcgtggccg cgctgcgatc cagtgcgagt gttgctacta cgcagaggaa gctacagcga 60
gaccgccact agatttaggg gacggcgcgc cgcgagggca cgccgatccc caacaccaag 120
cccgggggct tgagggttga aatgacgctc gaacaggcat gcccgccaga atactggcgg 180
gcgcaatgtg cgttcaaaga ttcgatgatt cactgaattc tgcaattcac attacttatc 240
gcatttcgct gcgttcttca tcgatgccag aaccaagaga tccgttgttg aaagttttga 300
tttatttaat cgtgttactc agaagatact gtataaaaca agagtttggg ggtcctctgg 360
cgggccgccg gagcgggcac cgccgaggca acgataggta tgttcacagg ggtttgggag 420
ttgtaaactc ggtaatgatc cctccgctgg ttcaccaacg gagaccttgt tacgactttt 480
acttcctc 488

Claims (9)

  1. The fungus of the genus Neonectria is Neonectria fungus (Neonectria sp.) CPCC401225, and the accession number of the Neonectria fungus (Neonectria sp.) CPCC401225 in the China general microbiological culture Collection center is CGMCC No. 20277.
  2. 2. A culture of a fungus belonging to the genus Neonectria as defined in claim 1, which is obtained by culturing a fungus belonging to the genus Neonectria (Neonectria sp.) CPCC401225 as defined in claim 1 in a microbial culture medium.
  3. 3. A microbial preparation comprising the fungus belonging to the genus Neonectria of claim 1 or/and the culture of claim 2.
  4. 4. The microbial inoculum of claim 3, wherein: the microbial inoculum has at least one of the following functions:
    a1, treating and/or preventing diseases caused by staphylococcus aureus,
    a2, treating and/or preventing diseases caused by enterococcus faecium,
    a3, treatment and/or prevention of diseases caused by Escherichia coli,
    a4, treating and/or preventing diseases caused by Klebsiella pneumoniae,
    a5, treating and/or preventing diseases caused by Acinetobacter baumannii,
    a6, inhibiting the growth of staphylococcus aureus,
    a7, inhibiting the growth of enterococcus faecium,
    a8, inhibiting the growth of Escherichia coli,
    a9, inhibiting the growth of Klebsiella pneumoniae,
    a10, inhibiting the growth of acinetobacter baumannii.
  5. 5. A product containing the Neonectria fungus CPCC401225 as defined in claim 1, the culture as defined in claim 2, or the microbial agent as defined in claim 3 or 4, said product being a pharmaceutical, a food product, a nutraceutical, an animal feed, or an animal feed additive.
  6. 6. The product of claim 5, wherein: the product has at least one of the following functions:
    a1, treating and/or preventing diseases caused by staphylococcus aureus,
    a2, treating and/or preventing diseases caused by enterococcus faecium,
    a3, treatment and/or prevention of diseases caused by Escherichia coli,
    a4, treating and/or preventing diseases caused by Klebsiella pneumoniae,
    a5, treating and/or preventing diseases caused by Acinetobacter baumannii,
    a6, inhibiting the growth of staphylococcus aureus,
    a7, inhibiting the growth of enterococcus faecium,
    a8, inhibiting the growth of Escherichia coli,
    a9, inhibiting the growth of Klebsiella pneumoniae,
    a10, inhibiting the growth of acinetobacter baumannii.
  7. 7. Use of the fungus CPCC401225 belonging to the genus Neonectria of claim 1, or the culture of claim 2, or the bacterial agent of claim 3 or 4 for the preparation of a functional product selected from the group consisting of:
    a1, treating and/or preventing diseases caused by staphylococcus aureus,
    a2, treating and/or preventing diseases caused by enterococcus faecium,
    a3, treatment and/or prevention of diseases caused by Escherichia coli,
    a4, treating and/or preventing diseases caused by Klebsiella pneumoniae,
    a5, treating and/or preventing diseases caused by Acinetobacter baumannii,
    a6, inhibiting the growth of staphylococcus aureus,
    a7, inhibiting the growth of enterococcus faecium,
    a8, inhibiting the growth of Escherichia coli,
    a9, inhibiting the growth of Klebsiella pneumoniae,
    a10, inhibiting the growth of acinetobacter baumannii.
  8. 8. Use of the Neonectria fungus CPCC401225 of claim 1, the culture of claim 2, or the inoculant of claim 3 or 4 for the preparation of Compound 1, Compound 2 and/or Compound 3, wherein Compound 1 is represented by formula I, Compound 2 is represented by formula II, and Compound 3 is represented by formula III:
    Figure FDA0003683919050000021
    Figure FDA0003683919050000031
  9. 9. a method of preparing a compound comprising: culturing a fungus of the genus Neonectria in a culture medium to obtain a fermentation product, and obtaining the compound from the fermentation product; the Neonectria fungus is Neonectria fungus (Neonectria sp.) CPCC401225, which is registered with CGMCC No.20277 in China general microbiological culture Collection center; the structural formula of the compound is shown as formula I or II or III:
    Figure FDA0003683919050000032
    Figure FDA0003683919050000041
CN202110555886.6A 2021-05-21 2021-05-21 Gram-positive bacterium resisting compound derived from polar region source fungi as well as preparation method and application thereof Active CN113249231B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN202110555886.6A CN113249231B (en) 2021-05-21 2021-05-21 Gram-positive bacterium resisting compound derived from polar region source fungi as well as preparation method and application thereof

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN202110555886.6A CN113249231B (en) 2021-05-21 2021-05-21 Gram-positive bacterium resisting compound derived from polar region source fungi as well as preparation method and application thereof

Publications (2)

Publication Number Publication Date
CN113249231A CN113249231A (en) 2021-08-13
CN113249231B true CN113249231B (en) 2022-08-02

Family

ID=77183511

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202110555886.6A Active CN113249231B (en) 2021-05-21 2021-05-21 Gram-positive bacterium resisting compound derived from polar region source fungi as well as preparation method and application thereof

Country Status (1)

Country Link
CN (1) CN113249231B (en)

Family Cites Families (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2007127218A2 (en) * 2006-04-26 2007-11-08 Taro Pharmaceuticals U.S.A., Inc. Streptomyces-derived antimicrobial compound and method of using same against antibiotic-resistant bacteria
ES2368236B1 (en) * 2009-12-23 2012-09-26 Merck Sharp & Dohme Corp. ANTIBACTERIAL AGENTS.
KR101764349B1 (en) * 2011-01-04 2017-08-03 한국생명공학연구원 A novel flavimycin compound having peptide deformylayse inhibition and antibacterial activity
WO2012093859A2 (en) * 2011-01-04 2012-07-12 한국생명공학연구원 Antifungal composition including polycyclic peptide compound and method for producing same
CN103565798B (en) * 2012-07-30 2015-12-23 成都中医药大学 The novelty teabag of Pogostone and derivant thereof
UA126438C2 (en) * 2018-12-27 2022-10-05 Ігор Семенович Марков Polyvalent combined inactivated liquid vaccine against bacterial and fungal pathogens of the nasopharynx, mouth and eyes, a method of its manufacture and a method of treatment and prevention thereof
UA126440C2 (en) * 2018-12-27 2022-10-05 Ігор Семенович Марков Polyvalent combined inactivated liquid vaccine against pathogens of bacterial and fungal diseases of the genitourinary system, a method of its manufacture and a method of treatment and prevention thereof
CN111394361B (en) * 2018-12-27 2022-04-26 中粮生物科技（北京）有限公司 Brevibacillus laterosporus, composition and application thereof
KR102225240B1 (en) * 2019-07-03 2021-03-10 경북대학교 산학협력단 Use of novel recombinant protein Ablysin killing pathogenic multidrug resistant bacteria

Also Published As

Publication number Publication date
CN113249231A (en) 2021-08-13

Similar Documents

Publication Publication Date Title
CN111548954B (en) Four anthraquinone compounds and preparation method and application thereof
CN115851500B (en) Lactobacillus plantarum and application thereof
JPH04217683A (en) Antibiotic
CN113249231B (en) Gram-positive bacterium resisting compound derived from polar region source fungi as well as preparation method and application thereof
JPH04217697A (en) Antibiotics
CN111848558B (en) Anti-influenza virus compound derived from Talaromyces, and preparation method and application thereof
US8003602B2 (en) Antibacterial compounds
US20100227918A1 (en) Streptomyces-derived antimicrobial compound and method of using same against antibiotic-resistant bacteria
CN116144503A (en) Fungus of genus Subplenodonomus, antibacterial compound, preparation method and application thereof
CN115109708B (en) Fungus and antibacterial compound with photinia and preparation method and application thereof
JP2008513486A (en) Antibiotic compound
RU2444526C2 (en) Novel antibacterial compounds
TW202031257A (en) Metabolite of lactic acid bacteria and use of the metabolite of lactic acid bacteria for protecting lungs
CN110698537B (en) Natural Rakicidins compound Rakicidin B1-2 and fermentation extraction method thereof
CN103304628B (en) Nosiheptide derivative and uses thereof
JP5816196B2 (en) Extract from Kibdelosporangium as antibacterial agent
CN107652262B (en) Preparation method and application of amicumacecin isocoumarin compound
CN116240252A (en) Trideceth derivative and preparation method and application thereof
CN116875561A (en) Phage, preparation and application thereof
WO2007127293A2 (en) Novel streptomyces-derived antimicrobial compound and method of using same against antibiotic-resistant bacteria
CN111533756A (en) Spiroketal compound derived from plant pathogenic fungi, preparation method and application
US20100144658A1 (en) Novel antibiotics, bispolides a1, a2, and a3 as well as bispolides b1, b2a, b2b and b3 and processes for producing said antibiotics
JPH03209384A (en) Novel microbial transformation product

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