CN116196324A - Composition of pyrolin and clindamycin and application thereof in bacteria inhibition - Google Patents
Composition of pyrolin and clindamycin and application thereof in bacteria inhibition Download PDFInfo
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- CN116196324A CN116196324A CN202310071226.XA CN202310071226A CN116196324A CN 116196324 A CN116196324 A CN 116196324A CN 202310071226 A CN202310071226 A CN 202310071226A CN 116196324 A CN116196324 A CN 116196324A
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- pyrolin
- clindamycin
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- streptococcus suis
- escherichia coli
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- CNHDIAIOKMXOLK-UHFFFAOYSA-N toluquinol Chemical group CC1=CC(O)=CC=C1O CNHDIAIOKMXOLK-UHFFFAOYSA-N 0.000 title claims abstract description 160
- 229960002227 clindamycin Drugs 0.000 title claims abstract description 62
- KDLRVYVGXIQJDK-AWPVFWJPSA-N clindamycin Chemical compound CN1C[C@H](CCC)C[C@H]1C(=O)N[C@H]([C@H](C)Cl)[C@@H]1[C@H](O)[C@H](O)[C@@H](O)[C@@H](SC)O1 KDLRVYVGXIQJDK-AWPVFWJPSA-N 0.000 title claims abstract description 62
- 239000000203 mixture Substances 0.000 title claims abstract description 21
- 230000005764 inhibitory process Effects 0.000 title abstract description 15
- 241000894006 Bacteria Species 0.000 title abstract description 10
- 241000194021 Streptococcus suis Species 0.000 claims abstract description 37
- 241000588724 Escherichia coli Species 0.000 claims abstract description 34
- 241000606748 Actinobacillus pleuropneumoniae Species 0.000 claims abstract description 33
- 241000606807 Glaesserella parasuis Species 0.000 claims abstract description 33
- 239000003814 drug Substances 0.000 claims abstract description 30
- 230000002401 inhibitory effect Effects 0.000 claims description 15
- 238000000338 in vitro Methods 0.000 claims description 10
- 208000015181 infectious disease Diseases 0.000 claims description 7
- 241000736246 Pyrola Species 0.000 claims description 3
- 206010061372 Streptococcal infection Diseases 0.000 claims description 3
- 206010061126 Escherichia infection Diseases 0.000 claims description 2
- 241000194017 Streptococcus Species 0.000 claims description 2
- 208000020612 escherichia coli infection Diseases 0.000 claims description 2
- 241000606750 Actinobacillus Species 0.000 claims 1
- 244000025254 Cannabis sativa Species 0.000 claims 1
- 241000606790 Haemophilus Species 0.000 claims 1
- 230000000844 anti-bacterial effect Effects 0.000 abstract description 37
- 229940079593 drug Drugs 0.000 abstract description 18
- 230000002195 synergetic effect Effects 0.000 abstract description 8
- 238000011161 development Methods 0.000 abstract description 3
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- 238000009395 breeding Methods 0.000 abstract 1
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- 238000002474 experimental method Methods 0.000 description 4
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- QIGBRXMKCJKVMJ-UHFFFAOYSA-N Hydroquinone Chemical compound OC1=CC=C(O)C=C1 QIGBRXMKCJKVMJ-UHFFFAOYSA-N 0.000 description 2
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- 239000003153 chemical reaction reagent Substances 0.000 description 2
- MYSWGUAQZAJSOK-UHFFFAOYSA-N ciprofloxacin Chemical compound C12=CC(N3CCNCC3)=C(F)C=C2C(=O)C(C(=O)O)=CN1C1CC1 MYSWGUAQZAJSOK-UHFFFAOYSA-N 0.000 description 2
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- UCSJYZPVAKXKNQ-HZYVHMACSA-N streptomycin Chemical compound CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](NC(N)=N)[C@H](O)[C@@H](NC(N)=N)[C@H](O)[C@H]1O UCSJYZPVAKXKNQ-HZYVHMACSA-N 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 229960004885 tiamulin Drugs 0.000 description 2
- UURAUHCOJAIIRQ-QGLSALSOSA-N tiamulin Chemical compound CCN(CC)CCSCC(=O)O[C@@H]1C[C@@](C)(C=C)[C@@H](O)[C@H](C)[C@@]23CC[C@@H](C)[C@]1(C)[C@@H]2C(=O)CC3 UURAUHCOJAIIRQ-QGLSALSOSA-N 0.000 description 2
- 229960005486 vaccine Drugs 0.000 description 2
- LITBAYYWXZOHAW-XDZRHBBOSA-N (2s,5r,6r)-6-[[(2r)-2-[(4-ethyl-2,3-dioxopiperazine-1-carbonyl)amino]-2-phenylacetyl]amino]-3,3-dimethyl-7-oxo-4-thia-1-azabicyclo[3.2.0]heptane-2-carboxylic acid;(2s,3s,5r)-3-methyl-4,4,7-trioxo-3-(triazol-1-ylmethyl)-4$l^{6}-thia-1-azabicyclo[3.2.0]hept Chemical compound C([C@]1(C)S([C@H]2N(C(C2)=O)[C@H]1C(O)=O)(=O)=O)N1C=CN=N1.O=C1C(=O)N(CC)CCN1C(=O)N[C@H](C=1C=CC=CC=1)C(=O)N[C@@H]1C(=O)N2[C@@H](C(O)=O)C(C)(C)S[C@@H]21 LITBAYYWXZOHAW-XDZRHBBOSA-N 0.000 description 1
- KMEGBUCIGMEPME-LQYKFRDPSA-N (2s,5r,6r)-6-[[(2r)-2-amino-2-phenylacetyl]amino]-3,3-dimethyl-7-oxo-4-thia-1-azabicyclo[3.2.0]heptane-2-carboxylic acid;(1r,4s)-3,3-dimethyl-2,2,6-trioxo-2$l^{6}-thiabicyclo[3.2.0]heptane-4-carboxylic acid Chemical compound O=S1(=O)C(C)(C)[C@H](C(O)=O)C2C(=O)C[C@H]21.C1([C@@H](N)C(=O)N[C@H]2[C@H]3SC([C@@H](N3C2=O)C(O)=O)(C)C)=CC=CC=C1 KMEGBUCIGMEPME-LQYKFRDPSA-N 0.000 description 1
- 108010077805 Bacterial Proteins Proteins 0.000 description 1
- 208000035143 Bacterial infection Diseases 0.000 description 1
- 241000606125 Bacteroides Species 0.000 description 1
- 241001070941 Castanea Species 0.000 description 1
- 235000014036 Castanea Nutrition 0.000 description 1
- 206010012735 Diarrhoea Diseases 0.000 description 1
- 241000194032 Enterococcus faecalis Species 0.000 description 1
- 235000007297 Gaultheria procumbens Nutrition 0.000 description 1
- 229930182566 Gentamicin Natural products 0.000 description 1
- CEAZRRDELHUEMR-URQXQFDESA-N Gentamicin Chemical compound O1[C@H](C(C)NC)CC[C@@H](N)[C@H]1O[C@H]1[C@H](O)[C@@H](O[C@@H]2[C@@H]([C@@H](NC)[C@@](C)(O)CO2)O)[C@H](N)C[C@@H]1N CEAZRRDELHUEMR-URQXQFDESA-N 0.000 description 1
- 102000013691 Interleukin-17 Human genes 0.000 description 1
- 108050003558 Interleukin-17 Proteins 0.000 description 1
- GSDSWSVVBLHKDQ-JTQLQIEISA-N Levofloxacin Chemical compound C([C@@H](N1C2=C(C(C(C(O)=O)=C1)=O)C=C1F)C)OC2=C1N1CCN(C)CC1 GSDSWSVVBLHKDQ-JTQLQIEISA-N 0.000 description 1
- 201000009906 Meningitis Diseases 0.000 description 1
- 206010034133 Pathogen resistance Diseases 0.000 description 1
- 241000228143 Penicillium Species 0.000 description 1
- 206010035664 Pneumonia Diseases 0.000 description 1
- 108010093965 Polymyxin B Proteins 0.000 description 1
- 206010040047 Sepsis Diseases 0.000 description 1
- 241000191963 Staphylococcus epidermidis Species 0.000 description 1
- 241000194048 Streptococcus equi Species 0.000 description 1
- WKDDRNSBRWANNC-UHFFFAOYSA-N Thienamycin Natural products C1C(SCCN)=C(C(O)=O)N2C(=O)C(C(O)C)C21 WKDDRNSBRWANNC-UHFFFAOYSA-N 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 230000001154 acute effect Effects 0.000 description 1
- 230000000996 additive effect Effects 0.000 description 1
- 229960004821 amikacin Drugs 0.000 description 1
- LKCWBDHBTVXHDL-RMDFUYIESA-N amikacin Chemical compound O([C@@H]1[C@@H](N)C[C@H]([C@@H]([C@H]1O)O[C@@H]1[C@@H]([C@@H](N)[C@H](O)[C@@H](CO)O1)O)NC(=O)[C@@H](O)CCN)[C@H]1O[C@H](CN)[C@@H](O)[C@H](O)[C@H]1O LKCWBDHBTVXHDL-RMDFUYIESA-N 0.000 description 1
- 229940126575 aminoglycoside Drugs 0.000 description 1
- 229940038195 amoxicillin / clavulanate Drugs 0.000 description 1
- 229960000723 ampicillin Drugs 0.000 description 1
- AVKUERGKIZMTKX-NJBDSQKTSA-N ampicillin Chemical compound C1([C@@H](N)C(=O)N[C@H]2[C@H]3SC([C@@H](N3C2=O)C(O)=O)(C)C)=CC=CC=C1 AVKUERGKIZMTKX-NJBDSQKTSA-N 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000003042 antagnostic effect Effects 0.000 description 1
- 230000008485 antagonism Effects 0.000 description 1
- 230000003064 anti-oxidating effect Effects 0.000 description 1
- 230000002155 anti-virotic effect Effects 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 230000000975 bioactive effect Effects 0.000 description 1
- 230000008827 biological function Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 229960002100 cefepime Drugs 0.000 description 1
- HVFLCNVBZFFHBT-ZKDACBOMSA-N cefepime Chemical compound S([C@@H]1[C@@H](C(N1C=1C([O-])=O)=O)NC(=O)\C(=N/OC)C=2N=C(N)SC=2)CC=1C[N+]1(C)CCCC1 HVFLCNVBZFFHBT-ZKDACBOMSA-N 0.000 description 1
- 229960004261 cefotaxime Drugs 0.000 description 1
- GPRBEKHLDVQUJE-VINNURBNSA-N cefotaxime Chemical compound N([C@@H]1C(N2C(=C(COC(C)=O)CS[C@@H]21)C(O)=O)=O)C(=O)/C(=N/OC)C1=CSC(N)=N1 GPRBEKHLDVQUJE-VINNURBNSA-N 0.000 description 1
- 229960000484 ceftazidime Drugs 0.000 description 1
- NMVPEQXCMGEDNH-TZVUEUGBSA-N ceftazidime pentahydrate Chemical compound O.O.O.O.O.S([C@@H]1[C@@H](C(N1C=1C([O-])=O)=O)NC(=O)\C(=N/OC(C)(C)C(O)=O)C=2N=C(N)SC=2)CC=1C[N+]1=CC=CC=C1 NMVPEQXCMGEDNH-TZVUEUGBSA-N 0.000 description 1
- 229960004755 ceftriaxone Drugs 0.000 description 1
- VAAUVRVFOQPIGI-SPQHTLEESA-N ceftriaxone Chemical compound S([C@@H]1[C@@H](C(N1C=1C(O)=O)=O)NC(=O)\C(=N/OC)C=2N=C(N)SC=2)CC=1CSC1=NC(=O)C(=O)NN1C VAAUVRVFOQPIGI-SPQHTLEESA-N 0.000 description 1
- 229960005091 chloramphenicol Drugs 0.000 description 1
- WIIZWVCIJKGZOK-RKDXNWHRSA-N chloramphenicol Chemical compound ClC(Cl)C(=O)N[C@H](CO)[C@H](O)C1=CC=C([N+]([O-])=O)C=C1 WIIZWVCIJKGZOK-RKDXNWHRSA-N 0.000 description 1
- 229960003405 ciprofloxacin Drugs 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 229940032049 enterococcus faecalis Drugs 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 229960002518 gentamicin Drugs 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 229960002182 imipenem Drugs 0.000 description 1
- ZSKVGTPCRGIANV-ZXFLCMHBSA-N imipenem Chemical compound C1C(SCC\N=C\N)=C(C(O)=O)N2C(=O)[C@H]([C@H](O)C)[C@H]21 ZSKVGTPCRGIANV-ZXFLCMHBSA-N 0.000 description 1
- 230000002147 killing effect Effects 0.000 description 1
- 229960003376 levofloxacin Drugs 0.000 description 1
- 239000003120 macrolide antibiotic agent Substances 0.000 description 1
- 229940041033 macrolides Drugs 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- OSWPMRLSEDHDFF-UHFFFAOYSA-N methyl salicylate Chemical compound COC(=O)C1=CC=CC=C1O OSWPMRLSEDHDFF-UHFFFAOYSA-N 0.000 description 1
- 239000006994 mh medium Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000008506 pathogenesis Effects 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 229920000024 polymyxin B Polymers 0.000 description 1
- 229960005266 polymyxin b Drugs 0.000 description 1
- -1 polyphenol compound Chemical class 0.000 description 1
- 235000013824 polyphenols Nutrition 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- DWHGNUUWCJZQHO-ZVDZYBSKSA-M potassium;(2s,5r,6r)-6-[[(2r)-2-amino-2-(4-hydroxyphenyl)acetyl]amino]-3,3-dimethyl-7-oxo-4-thia-1-azabicyclo[3.2.0]heptane-2-carboxylic acid;(2r,3z,5r)-3-(2-hydroxyethylidene)-7-oxo-4-oxa-1-azabicyclo[3.2.0]heptane-2-carboxylate Chemical compound [K+].[O-]C(=O)[C@H]1C(=C/CO)/O[C@@H]2CC(=O)N21.C1([C@@H](N)C(=O)N[C@H]2[C@H]3SC([C@@H](N3C2=O)C(O)=O)(C)C)=CC=C(O)C=C1 DWHGNUUWCJZQHO-ZVDZYBSKSA-M 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 238000007619 statistical method Methods 0.000 description 1
- 229960005322 streptomycin Drugs 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 229940124530 sulfonamide Drugs 0.000 description 1
- 150000003456 sulfonamides Chemical class 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 229940040944 tetracyclines Drugs 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 244000000023 zoonotic pathogen Species 0.000 description 1
Classifications
-
- 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
- A61K31/045—Hydroxy compounds, e.g. alcohols; Salts thereof, e.g. alcoholates
- A61K31/05—Phenols
-
- 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
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7042—Compounds having saccharide radicals and heterocyclic rings
- A61K31/7052—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides
- A61K31/7056—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing five-membered rings with nitrogen as a ring hetero atom
-
- 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
-
- 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
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Veterinary Medicine (AREA)
- Chemical & Material Sciences (AREA)
- Public Health (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Organic Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- General Chemical & Material Sciences (AREA)
- Oncology (AREA)
- Molecular Biology (AREA)
- Communicable Diseases (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention belongs to the technical field of veterinary medicines, and discloses a pyrolin and clindamycin composition and application thereof in bacteria inhibition. The composition of the pyrolin and the clindamycin has synergistic antibacterial effect on streptococcus suis, escherichia coli, actinobacillus pleuropneumoniae and haemophilus parasuis, reduces the concentration of independent antibacterial effect of the pyrolin and the clindamycin, saves the cost, and has wide application prospect and development value in the fields of food processing, medicine and breeding industry.
Description
Technical Field
The invention belongs to the technical field of veterinary medicines, and particularly relates to a pyrolin and clindamycin composition and application thereof in bacteria inhibition.
Background
Streptococcus Suis (SS) is an important zoonotic pathogen, and can be classified into 33 serotypes according to the difference of capsular polysaccharides, and can infect pigs and humans, causing sepsis, meningitis, pneumonia, arthritis and even death. Actinobacillus pleuropneumoniae (App) has now found 15 serotypes, some of which are non-pathogenic and some of which can cause severe disease. Coli (E.coli) K88, K99, 987P strains often cause diarrhea in newborn piglets within 1 week, and their onset is characterized by acute onset, high mortality, and poor therapeutic effect after onset. Haemophilus parasuis is a contact bacterial infectious disease of pigs caused by haemophilus parasuis, and is also called as multiple serositis and arthritis of pigs, grignard disease and fibrinous serositis of pigs. At present, the focus of bacterial infection control is mainly prevention and antibiotic treatment. However, with the large and unreasonable use of antibiotics, bacterial resistance is very serious.
Clinical trial data shows that different degrees of resistance appear in different countries and regions of isolated streptococcus suis. In the Italian Cucco isolated Streptococcus suis, 61.5% of the strains developed resistance to two antibiotics and 29.5% of the strains developed resistance to three antibiotics. Canadian scholars Arndt tested 379 strains of Streptococcus suis for resistance to tetracycline, tiamulin and spectinomycin, and found that most strains were resistant to tetracycline, tiamulin and spectinomycin. Among 1163 strains of streptococcus suis isolated clinically from vanHout, the netherlands scholars had 78.4% resistance to tetracycline and 48.1% resistance to clindamycin. The clinical 34 strains of streptococcus suis are subjected to drug resistance analysis by Chinese scholars, 91.18 percent of strains are multi-drug resistant, 82.3 to 100.0 percent of strains are drug resistant to sulfonamides, 79.4 to 94.1 percent of strains are drug resistant to aminoglycosides, 88.2 to 97.1 percent of strains are drug resistant to tetracyclines, 79.4 to 85.3 percent of linkeamine drugs are drug resistant, and 50 to 88.2 percent of strains are drug resistant to macrolides. The Chinese scholars carry out drug resistance detection on 154 strains of escherichia coli, 151 strains (98%) are multi-drug resistant bacteria, the drug resistance rate of the escherichia coli to compound neonomine, tetracycline, ampicillin, streptomycin and chloramphenicol is 81% -100%, the drug resistance rate of the amoxicillin/clavulanic acid, cefotaxime, gentamicin, ceftriaxone, ciprofloxacin and amikacin is 31% -66%, and the drug resistance rate of the amoxicillin/clavulanate to levofloxacin, polymyxin B, ceftazidime, cefepime, ampicillin-sulbactam, piperacillin-tazobactam and imipenem is 1% -19%. Thus, searching for new antibiotic substitutes, developing new antibacterial agents, or finding compounds that reverse antibiotic resistance are key approaches to solving the antibiotic resistance problem.
Pyrolin (molecular weight 124.13, molecular formula: C) 7 H 8 O 2 ) Is a natural polyphenol compound, is a representative hydroquinone substance extracted from wintergreen, and has biological functions of antivirus, antibacterial, antioxidation and the like. At present, the literature reports that pyrolin has better antibacterial effect on staphylococcus aureus and escherichia coli, poorer antibacterial effect on pseudomonas aeruginosa, better effect on common putrefying mycorrhizal fungi and penicillium in food, but poorer effect on brown rot fungi and chestnut epidemic fungi (the antibacterial effect of pyrola extract and application research [ D ] of Wangjiyan)]University of southwest, 2007). It is speculated that Pseudomonas aeruginosa is insensitive to pyrolin.
Clindamycin is a linkeamine antibiotic, and the main antibacterial action mechanism is combined with a bacterial ribosome 50S subunit to inhibit peptide chain extension, so that synthesis of bacterial proteins is inhibited. Clindamycin is commonly used to treat infections of bacteria including staphylococcus aureus, staphylococcus epidermidis, streptococcus (except enterococcus faecalis), pneumococcus, bacteroides and the like.
Disclosure of Invention
The invention aims to solve the technical problems of serious drug resistance of the existing streptococcus suis, escherichia coli, actinobacillus pleuropneumoniae and haemophilus parasuis and provides a pyrolin composition, which comprises the following components: pyrolin and clindamycin.
Another problem of the present invention is to provide the use of a pyrolin composition for the preparation of a medicament for the treatment or prophylaxis of infection by streptococcus suis, escherichia coli, haemophilus parasuis, or actinobacillus pleuropneumoniae. The composition provided by the invention has good antibacterial and bactericidal effects on streptococcus suis, escherichia coli, haemophilus parasuis and actinobacillus pleuropneumoniae.
In order to achieve the above purpose, the present invention adopts the following technical scheme:
use of a pyrolin composition in the manufacture of a medicament for the treatment or prophylaxis of infection by streptococcus suis (Streptococcus suis), escherichia coli, haemophilus parasuis (Haemophilus parasuis), or actinobacillus pleuropneumoniae (Actinobacillus pleuropneumoniae), the only active ingredients of said pyrolin composition being pyrolin and clindamycin.
In the above application, preferably, when used for preparing a medicament for treating or preventing streptococcus suis infection, the mass ratio of pyrolin to clindamycin is 1:0.125-32;
in the above application, preferably, when used for preparing a medicament for treating or preventing an escherichia coli infection, the mass ratio of pyrolin to clindamycin is 1:16-32;
in the above application, preferably, when used for preparing a medicament for treating or preventing haemophilus parasuis infection, the mass ratio of pyrolin to clindamycin is 1:8;
in the above application, preferably, the mass ratio of pyrolin to clindamycin is 1:8 when used for preparing a medicament for treating or preventing actinobacillus pleuropneumoniae infection.
The protection scope of the invention also comprises: the application of the pyrolin composition in inhibiting streptococcus suis, escherichia coli, haemophilus parasuis or actinobacillus pleuropneumoniae in vitro is provided, wherein the only effective components of the pyrolin composition are pyrolin and clindamycin.
In the above application, preferably, when used for in vitro inhibition of streptococcus suis, the mass ratio of pyrolin to clindamycin is 1:0.125-32;
in the above application, preferably, when used for in vitro inhibition of E.coli, the mass ratio of pyrolin to clindamycin is 1:16-32;
in the above application, preferably, when used for in vitro inhibition of haemophilus parasuis, the mass ratio of pyrolin to clindamycin is 1:8;
in the above application, it is preferred that the mass ratio of pyrolin to clindamycin is 1:8 when used for in vitro inhibition of actinobacillus pleuropneumoniae.
Compared with the prior art, the invention has the following advantages and effects:
the invention provides a synergistic antibacterial effect of pyrolin and clindamycin for the first time, and the composition can be used for resisting streptococcus suis, escherichia coli, actinobacillus pleuropneumoniae and haemophilus parasuis. The invention proves that the pyrolin and clindamycin composition has good antibacterial effect on streptococcus suis, escherichia coli, actinobacillus pleuropneumoniae and haemophilus parasuis through a plurality of method researches, lays a theoretical foundation for developing novel medicines for resisting streptococcus suis, escherichia coli, actinobacillus pleuropneumoniae and haemophilus parasuis diseases, provides candidate substances for the development of antibiotic substitutes, and has good application prospects in the aspects of preventing and treating streptococcus suis, escherichia coli, actinobacillus pleuropneumoniae and haemophilus parasuis diseases.
After the clindamycin and the pyrolin are combined, the antibacterial concentration of the clindamycin and the antibacterial concentration of the pyrolin are respectively reduced, and the index of the grading antibacterial concentration (FICI) of the combined medicament is 0.1875-0.5, and the clindamycin and the pyrolin have synergistic antibacterial effect (the synergistic antibacterial effect, the FICI is less than or equal to 0.5, the additive effect, the FICI is less than or equal to 0.5 and less than or equal to 1, the irrelevant effect, the FICI is less than or equal to 1 and less than or equal to 2, and the antagonistic effect, the FICI is more than 2). Therefore, the pyrolin can reverse the drug resistance of streptococcus suis, escherichia coli, actinobacillus pleuropneumoniae and haemophilus parasuis to clindamycin, and the clindamycin and pyrolin composition is expected to become novel bioactive molecules for preventing and treating streptococcus suis, escherichia coli, actinobacillus pleuropneumoniae and haemophilus parasuis diseases, provides a solution for preventing and treating drug-resistant streptococcus suis, escherichia coli, actinobacillus pleuropneumoniae and haemophilus parasuis, has good application prospect in preventing and treating streptococcus suis, escherichia coli, actinobacillus pleuropneumoniae and haemophilus parasuis diseases, and provides a theoretical basis for popularization and application of the pyrolin composition as an antibiotic substitute.
Detailed Description
The present invention is further illustrated below with reference to specific examples, which are not intended to limit the invention in any way. Unless specifically stated otherwise, the reagents, methods and apparatus employed in the present invention are those conventional in the art.
Reagents and materials used in the following examples are commercially available unless otherwise specified. The embodiment of the invention is illustrated by taking streptococcus suis (P1/7 strain, 29 strain, ATCC35246 strain and SC19 strain), escherichia coli (K88, K99 and 987P), actinobacillus pleuropneumoniae (4074 strain) and haemophilus parasuis (SH 0165 strain) as examples, and other streptococcus suis, escherichia coli, actinobacillus pleuropneumoniae and haemophilus parasuis have inhibiting and killing effects.
The structural formula of the pyrolin is as follows:
statistical analysis of the following examples of the invention: all experiments were repeated at least 3 times independently.
Example 1:
pyrolin was assayed for the role and mechanism of streptococcus suis (P1/7 strain, 29 strain (Xu Lei. IL-17A in streptococcus suis infection [ D ]. University of agriculture in chinese, 2022.doi: 10.27158/d.cnki.ghznu.2022.000333.), ATCC35246 strain, SC19 strain), escherichia coli (K88, K99, 987P), actinobacillus pleuropneumoniae (4074 strain) (methou, y., li, l., chen, z., yuan, h., chen, h., & Zhou, r. (2013). Adhesion protein ApfA of Actinobacillus pleuropneumoniae is required for pathogenesis and is a potential target for vaccine development and vaccine immunology: CVI,20 (2), 287-294), minimum inhibitory and minimum bactericidal concentrations of haemophilus parasuis (SH 0165 strain):
1. material
Mueller-Hinton (MH) broth (available from BD company) was used in drug susceptibility experiments to examine the bacteriostatic efficacy of drugs.
2. Test method
The purified Streptococcus suis (P1/7 strain, 29 strain, ATCC35246 strain, SC19 strain), escherichia coli (K88, K99, 987P), actinobacillus pleuropneumoniae (4074 strain), and haemophilus parasuis (SH 0165 strain) colonies were picked up by a sterile inoculating loop, and were sufficiently dispersed in MH liquid medium, each of which was prepared to be 5X 10 5 Standard bacterial suspension of CFU/ml;
pyrolactone solutions of different concentrations after dilution with MH broth were added to sterile 96-well polystyrene plates, respectively, with Pyrolactone solution at wells 1 to 10, 50. Mu.l per well, 50. Mu.l of MH broth at well 11 as growth control, and 100. Mu.l of MH broth at well 12 as negative control.
50 μl of standard bacterial suspension was added to wells 1 to 11, incubated at 37℃for 16-20h, and the MIC value was determined by observing the presence or absence of bacterial growth. The bacterial liquid was aspirated from each well, plated onto MH plates, and MBC was determined without colony growth.
3. Results
As shown in Table 1, pyrolin has antibacterial and bactericidal effects on Streptococcus suis (P1/7 strain, 29 strain, ATCC35246 strain, SC19 strain), escherichia coli (K88, K99, 987P), actinobacillus pleuropneumoniae (4074 strain), and haemophilus parasuis (SH 0165 strain), as shown in Table 1.
Table 1 minimum inhibitory and minimum bactericidal concentrations of pyrolin against bacteria
Example 2:
clindamycin was assayed for minimum inhibitory concentration and minimum bactericidal concentration of streptococcus suis (P1/7 strain, 29 strain, ATCC35246 strain, SC19 strain), escherichia coli (K88, K99, 987P), actinobacillus pleuropneumoniae (4074 strain), haemophilus parasuis (SH 0165 strain):
1. material
Mueller-Hinton (MH) broth (available from BD company) was used in drug susceptibility experiments to examine the bacteriostatic efficacy of drugs.
2. Test method
The purified Streptococcus suis (P1/7 strain, 29 strain, ATCC35246 strain, SC19 strain), escherichia coli (K88, K99, 987P), actinobacillus pleuropneumoniae (4074 strain), and haemophilus parasuis (SH 0165 strain) colonies were picked up by a sterile inoculating loop, and were sufficiently dispersed in MH liquid medium, each of which was prepared to be 5X 10 5 Standard bacterial suspension of CFU/ml;
clindamycin solutions of different concentrations after dilution with MH liquid medium in a multiple ratio were added to sterile 96-well polystyrene plates, respectively, with 50. Mu.l of each of the 1 st to 10 th Kong Jiake th clindamycin solutions, 50. Mu.l of MH liquid medium was added to each well, and 50. Mu.l of MH liquid medium was added to each 11 th well as a growth control, and 100. Mu.l of MH liquid medium alone was added to each 12 th well as a negative control.
50 μl of standard bacterial suspension was added to wells 1 to 11, incubated at 37℃for 16-20h, and the MIC value was determined by observing the presence or absence of bacterial growth. The bacterial liquid was aspirated from each well, plated onto MH plates, and MBC was determined without colony growth.
3. Results
As is clear from Table 2, clindamycin has antibacterial and bactericidal effects on Streptococcus suis (P1/7 strain, 29 strain, ATCC35246 strain, SC19 strain), escherichia coli (K88, K99, 987P), actinobacillus pleuropneumoniae (4074 strain), and Haemophilus parasuis (SH 0165 strain), but has weak antibacterial and bactericidal effects on some strains.
TABLE 2 minimum inhibitory and minimum bactericidal concentrations of clindamycin on bacteria
Example 3:
synergistic bacteriostatic action of the combination of pyrolin and clindamycin on Streptococcus suis (P1/7 strain, 29 strain, ATCC35246 strain, SC19 strain), escherichia coli (K88, K99, 987P), actinobacillus pleuropneumoniae (4074 strain), haemophilus parasuis (SH 0165 strain):
1. material
Mueller-Hinton (MH) broth (available from BD company) was used in drug susceptibility experiments to examine the bacteriostatic efficacy of drugs.
2. Test method
The purified streptococcus suis (P1/7 strain, 29 strain, ATCC35246 strain, SC19 strain), escherichia coli (K88, K99, 987P), actinobacillus pleuropneumoniae (4074 strain), haemophilus parasuis (SH 0165 strain) colonies are respectively picked up by a sterile inoculating loop, transferred after shaking overnight, and fully dispersed in MH liquid culture medium to prepare 5×10 5 Standard bacterial suspension of CFU/ml;
50 μl of MH liquid culture medium is added to each row of 2 to 10 holes of a sterile 96-hole polystyrene plate by a row gun, 4-time MIC concentration pyrolin solution is added to the 1 st hole and the 2 nd hole, 50 μl of pyrolin solution is sucked out of the second hole and added to the third hole after being blown and mixed uniformly by the row gun, and the like, the mixture is blown and mixed uniformly and diluted by a multiple ratio, and 50 μl of pyrolin solution is sucked out of the 11 holes and discarded. Wells 12 were filled with 50 μl MH broth as growth control, where each column contained uniform concentrations of pyrolin.
Different concentrations of clindamycin solutions were diluted in sterile 4ml EP tubes with MH broth according to MIC values of different strains for clindamycin, respectively, with a maximum concentration of 4 times MIC. Row 2 was half as concentrated as row 1, and so on, the concentration of 8 rows of added clindamycin was calculated and the corresponding solution was prepared, and then 50 μl of solution was added to 96 well polystyrene plates, with the antibiotic concentrations in each row being consistent.
At this time, each hole in the 96-hole polystyrene board contains clindamycin and pyrolin solutions with different concentrations, the concentration of clindamycin in each row of solutions is the same, and the concentrations of the 1 st row to the 8 th row are reduced in a multiple ratio; the concentrations of the pyrolin solutions contained in each column are the same, and the concentrations of the columns 1 to 10 are reduced in a multiple ratio.
Mu.l of standard bacterial suspension was added to each of wells 1 to 10 and 12, and after addition, 50. Mu.l or 100. Mu.l MH medium was added to each of 96 wells, so that the total system per well was 200. Mu.l. Each row of 11 th wells is a negative control, only pyrolin and clindamycin solution; wells 12 were growth control, standard bacterial fluid only.
And incubating the 96-well polystyrene plate for 16-20h at 37 ℃, observing whether bacteria grow or not, and calculating the synergistic antibacterial index according to the minimum combined antibacterial concentration of the antibiotics and the compounds.
The judgment method is as follows: synergistic antibacterial effect, FICI is less than or equal to 0.5; adding, wherein FICI is less than or equal to 1 and is more than 0.5; irrelevant, wherein FICI is 1< 2; antagonism, FICI >2.
3. Results
As can be seen from Table 3, when pyrolin was used in combination with clindamycin, the pyrolin concentration was 8. Mu.g/ml, the clindamycin concentration was 1. Mu.g/ml, which had an inhibitory effect on the growth of Streptococcus suis (P1/7), and the hierarchical inhibitory concentration index was 0.375; when the pyrolin is combined with clindamycin, the concentration of the pyrolin is 8 mug/ml, the concentration of the clindamycin is 256 mug/ml, the pyrolin has an inhibition effect on the growth of streptococcus suis (29), and the grading antibacterial concentration index is 0.25; the concentration of pyrolin is 16 mug/ml, the concentration of clindamycin is 4 mug/ml, the pyrolin has an inhibiting effect on the growth of streptococcus equi subspecies zooepidemicus (ATCC 35246), and the grading bacteriostatic concentration index is 0.375; the concentration of pyrolin is 16 mug/ml, the concentration of clindamycin is 16 mug/ml, the pyrolin has an inhibition effect on the growth of streptococcus suis (SC 19), and the grading antibacterial concentration index is 0.258; the concentration of pyrolin is 32 mug/ml, the concentration of clindamycin is 1024 mug/ml, the pyrolin has an inhibition effect on the growth of escherichia coli (K88), and the index of the grading antibacterial concentration is 0.5; the concentration of pyrolin is 32 mug/ml, the concentration of clindamycin is 512 mug/ml, the pyrolin has an inhibition effect on the growth of escherichia coli (K99), and the grading antibacterial concentration index is 0.25; the concentration of pyrolin is 32 mug/ml, the concentration of clindamycin is 512 mug/ml, the pyrolin has an inhibition effect on the growth of escherichia coli (987P), and the grading antibacterial concentration index is 0.25; the concentration of pyrolin is 8 mug/ml, the concentration of clindamycin is 64 mug/ml, the pyrolin has an inhibition effect on the growth of actinobacillus pleuropneumoniae (4074 strain), and the index of the grading antibacterial concentration is 0.1875; the concentration of pyrolin is 16 mug/ml, the concentration of clindamycin is 128 mug/ml, the pyrolin has an inhibition effect on the growth of haemophilus parasuis (SH 0165 strain), and the grading antibacterial concentration index is 0.375. The pyrolin and clindamycin have synergistic antibacterial effect. FICI calculation formula: MIC at MIC/a alone at MIC/a combination and MIC at MIC/B alone at mic+b combination at fici=a combination.
Table 3 minimum inhibitory concentration and graded inhibitory concentration index of pyrolin in combination with clindamycin on bacteria
The above examples are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above examples, and any other changes, modifications, substitutions, combinations, and simplifications that do not depart from the spirit and principle of the present invention should be made in the equivalent manner, and the embodiments are included in the protection scope of the present invention.
Claims (10)
1. Pyrola grass extract composition for treating or preventing streptococcus suisStreptococcus suis) Coli @Escherichia coli) Haemophilus parasuisHaemophilus parasuis) Or actinobacillus pleuropneumoniaeActinobacillus pleuropneumoniae) The only effective components of the pyrolin composition are pyrolin and clindamycin.
2. The use according to claim 1, characterized in that: when used for preparing the medicine for treating or preventing streptococcus suis infection, the mass ratio of the pyrolin to the clindamycin is 1:0.125-32.
3. The use according to claim 1, characterized in that: when used for preparing the medicine for treating or preventing the escherichia coli infection, the mass ratio of the pyrolin to the clindamycin is 1:16-32.
4. The use according to claim 1, characterized in that: when used for preparing the medicine for treating or preventing haemophilus parasuis infection, the mass ratio of the pyrolin to the clindamycin is 1:8.
5. The use according to claim 1, characterized in that: when used for preparing a medicament for treating or preventing actinobacillus pleuropneumoniae infection, the mass ratio of pyrolin to clindamycin is 1:8.
6. The application of the pyrolin composition in inhibiting streptococcus suis, escherichia coli, haemophilus parasuis or actinobacillus pleuropneumoniae in vitro is provided, wherein the only effective components of the pyrolin composition are pyrolin and clindamycin.
7. The use according to claim 6, characterized in that: when used for inhibiting streptococcus suis in vitro, the mass ratio of the pyrolin to the clindamycin is 1:0.125-32.
8. The use according to claim 6, characterized in that: when used for inhibiting escherichia coli in vitro, the mass ratio of the pyrolin to the clindamycin is 1:16-32.
9. The use according to claim 6, characterized in that: when used for inhibiting haemophilus parasuis in vitro, the mass ratio of pyrolin to clindamycin is 1:8.
10. The use according to claim 6, characterized in that: when used to inhibit actinobacillus pleuropneumoniae in vitro, the mass ratio of pyrolin to clindamycin is 1:8.
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