CN115745909A - Conjugate FST and preparation method and application thereof - Google Patents
Conjugate FST and preparation method and application thereof Download PDFInfo
- Publication number
- CN115745909A CN115745909A CN202211415973.2A CN202211415973A CN115745909A CN 115745909 A CN115745909 A CN 115745909A CN 202211415973 A CN202211415973 A CN 202211415973A CN 115745909 A CN115745909 A CN 115745909A
- Authority
- CN
- China
- Prior art keywords
- fst
- conjugate
- acid
- solvent
- florfenicol
- 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.)
- Granted
Links
- 238000002360 preparation method Methods 0.000 title claims abstract description 11
- 229960003760 florfenicol Drugs 0.000 claims abstract description 24
- AYIRNRDRBQJXIF-NXEZZACHSA-N (-)-Florfenicol Chemical compound CS(=O)(=O)C1=CC=C([C@@H](O)[C@@H](CF)NC(=O)C(Cl)Cl)C=C1 AYIRNRDRBQJXIF-NXEZZACHSA-N 0.000 claims abstract description 23
- 239000002904 solvent Substances 0.000 claims abstract description 23
- 238000006243 chemical reaction Methods 0.000 claims abstract description 15
- 241000894006 Bacteria Species 0.000 claims abstract description 14
- JGFZNNIVVJXRND-UHFFFAOYSA-N N,N-Diisopropylethylamine (DIPEA) Chemical compound CCN(C(C)C)C(C)C JGFZNNIVVJXRND-UHFFFAOYSA-N 0.000 claims abstract description 13
- KDYFGRWQOYBRFD-UHFFFAOYSA-L succinate(2-) Chemical compound [O-]C(=O)CCC([O-])=O KDYFGRWQOYBRFD-UHFFFAOYSA-L 0.000 claims abstract description 13
- 208000015181 infectious disease Diseases 0.000 claims abstract description 9
- JNMRHUJNCSQMMB-UHFFFAOYSA-N sulfathiazole Chemical compound C1=CC(N)=CC=C1S(=O)(=O)NC1=NC=CS1 JNMRHUJNCSQMMB-UHFFFAOYSA-N 0.000 claims abstract description 8
- 229960001544 sulfathiazole Drugs 0.000 claims abstract description 8
- 238000000034 method Methods 0.000 claims abstract description 6
- 238000003756 stirring Methods 0.000 claims abstract description 4
- 239000003814 drug Substances 0.000 claims description 18
- 150000003839 salts Chemical class 0.000 claims description 14
- 241000588724 Escherichia coli Species 0.000 claims description 13
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 8
- 241000191967 Staphylococcus aureus Species 0.000 claims description 8
- 208000035473 Communicable disease Diseases 0.000 claims description 7
- FALRKNHUBBKYCC-UHFFFAOYSA-N 2-(chloromethyl)pyridine-3-carbonitrile Chemical compound ClCC1=NC=CC=C1C#N FALRKNHUBBKYCC-UHFFFAOYSA-N 0.000 claims description 6
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 6
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 6
- MUBZPKHOEPUJKR-UHFFFAOYSA-N Oxalic acid Chemical compound OC(=O)C(O)=O MUBZPKHOEPUJKR-UHFFFAOYSA-N 0.000 claims description 6
- OFOBLEOULBTSOW-UHFFFAOYSA-N Propanedioic acid Natural products OC(=O)CC(O)=O OFOBLEOULBTSOW-UHFFFAOYSA-N 0.000 claims description 6
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 claims description 6
- 229940014800 succinic anhydride Drugs 0.000 claims description 6
- 241001465754 Metazoa Species 0.000 claims description 5
- 150000001875 compounds Chemical class 0.000 claims description 5
- VZCYOOQTPOCHFL-OWOJBTEDSA-N Fumaric acid Chemical compound OC(=O)\C=C\C(O)=O VZCYOOQTPOCHFL-OWOJBTEDSA-N 0.000 claims description 4
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 claims description 4
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 claims description 4
- 238000010992 reflux Methods 0.000 claims description 4
- VZCYOOQTPOCHFL-UHFFFAOYSA-N trans-butenedioic acid Natural products OC(=O)C=CC(O)=O VZCYOOQTPOCHFL-UHFFFAOYSA-N 0.000 claims description 4
- OISVCGZHLKNMSJ-UHFFFAOYSA-N 2,6-Lutidine Substances CC1=CC=CC(C)=N1 OISVCGZHLKNMSJ-UHFFFAOYSA-N 0.000 claims description 3
- -1 3H-1, 2, 3-triazolo [4,5-B ] pyridin-3-oxy Chemical group 0.000 claims description 3
- 241000282414 Homo sapiens Species 0.000 claims description 3
- 238000006555 catalytic reaction Methods 0.000 claims description 3
- 239000003085 diluting agent Substances 0.000 claims description 3
- BJEPYKJPYRNKOW-REOHCLBHSA-N (S)-malic acid Chemical compound OC(=O)[C@@H](O)CC(O)=O BJEPYKJPYRNKOW-REOHCLBHSA-N 0.000 claims description 2
- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 claims description 2
- CKLJMWTZIZZHCS-REOHCLBHSA-N L-aspartic acid Chemical compound OC(=O)[C@@H](N)CC(O)=O CKLJMWTZIZZHCS-REOHCLBHSA-N 0.000 claims description 2
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 claims description 2
- KDYFGRWQOYBRFD-UHFFFAOYSA-N Succinic acid Natural products OC(=O)CCC(O)=O KDYFGRWQOYBRFD-UHFFFAOYSA-N 0.000 claims description 2
- BJEPYKJPYRNKOW-UHFFFAOYSA-N alpha-hydroxysuccinic acid Natural products OC(=O)C(O)CC(O)=O BJEPYKJPYRNKOW-UHFFFAOYSA-N 0.000 claims description 2
- 229910000147 aluminium phosphate Inorganic materials 0.000 claims description 2
- 235000003704 aspartic acid Nutrition 0.000 claims description 2
- OQFSQFPPLPISGP-UHFFFAOYSA-N beta-carboxyaspartic acid Natural products OC(=O)C(N)C(C(O)=O)C(O)=O OQFSQFPPLPISGP-UHFFFAOYSA-N 0.000 claims description 2
- KDYFGRWQOYBRFD-NUQCWPJISA-N butanedioic acid Chemical compound O[14C](=O)CC[14C](O)=O KDYFGRWQOYBRFD-NUQCWPJISA-N 0.000 claims description 2
- 239000007795 chemical reaction product Substances 0.000 claims description 2
- 235000015165 citric acid Nutrition 0.000 claims description 2
- 239000003937 drug carrier Substances 0.000 claims description 2
- 239000001530 fumaric acid Substances 0.000 claims description 2
- 235000013922 glutamic acid Nutrition 0.000 claims description 2
- 239000004220 glutamic acid Substances 0.000 claims description 2
- VZCYOOQTPOCHFL-UPHRSURJSA-N maleic acid Chemical compound OC(=O)\C=C/C(O)=O VZCYOOQTPOCHFL-UPHRSURJSA-N 0.000 claims description 2
- 239000011976 maleic acid Substances 0.000 claims description 2
- 239000001630 malic acid Substances 0.000 claims description 2
- 235000011090 malic acid Nutrition 0.000 claims description 2
- 235000006408 oxalic acid Nutrition 0.000 claims description 2
- 239000000546 pharmaceutical excipient Substances 0.000 claims description 2
- 238000004519 manufacturing process Methods 0.000 claims 1
- 230000001954 sterilising effect Effects 0.000 abstract description 5
- 238000004659 sterilization and disinfection Methods 0.000 abstract description 5
- 229940124350 antibacterial drug Drugs 0.000 abstract description 3
- 244000052616 bacterial pathogen Species 0.000 abstract description 2
- 239000012295 chemical reaction liquid Substances 0.000 abstract 1
- 241000699670 Mus sp. Species 0.000 description 16
- 230000001580 bacterial effect Effects 0.000 description 13
- 229940079593 drug Drugs 0.000 description 10
- 239000000243 solution Substances 0.000 description 10
- 230000034994 death Effects 0.000 description 9
- 231100000517 death Toxicity 0.000 description 9
- 238000002474 experimental method Methods 0.000 description 9
- 239000001963 growth medium Substances 0.000 description 9
- 238000009631 Broth culture Methods 0.000 description 7
- 239000003242 anti bacterial agent Substances 0.000 description 7
- 229940088710 antibiotic agent Drugs 0.000 description 7
- 239000000725 suspension Substances 0.000 description 7
- 239000007788 liquid Substances 0.000 description 6
- 238000012360 testing method Methods 0.000 description 6
- VHYFNPMBLIVWCW-UHFFFAOYSA-N 4-Dimethylaminopyridine Chemical compound CN(C)C1=CC=NC=C1 VHYFNPMBLIVWCW-UHFFFAOYSA-N 0.000 description 4
- 238000005481 NMR spectroscopy Methods 0.000 description 4
- 230000036528 appetite Effects 0.000 description 4
- 235000019789 appetite Nutrition 0.000 description 4
- 238000001228 spectrum Methods 0.000 description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 4
- 238000001644 13C nuclear magnetic resonance spectroscopy Methods 0.000 description 3
- 238000005160 1H NMR spectroscopy Methods 0.000 description 3
- 206010012735 Diarrhoea Diseases 0.000 description 3
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- 238000007865 diluting Methods 0.000 description 3
- 239000007928 intraperitoneal injection Substances 0.000 description 3
- 238000001819 mass spectrum Methods 0.000 description 3
- 229960000549 4-dimethylaminophenol Drugs 0.000 description 2
- 208000035143 Bacterial infection Diseases 0.000 description 2
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- 206010059866 Drug resistance Diseases 0.000 description 2
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 2
- 239000006142 Luria-Bertani Agar Substances 0.000 description 2
- 208000022362 bacterial infectious disease Diseases 0.000 description 2
- 229910052799 carbon Inorganic materials 0.000 description 2
- 238000010168 coupling process Methods 0.000 description 2
- 230000007547 defect Effects 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 230000003203 everyday effect Effects 0.000 description 2
- 229910052739 hydrogen Inorganic materials 0.000 description 2
- 239000001257 hydrogen Substances 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 239000002689 soil Substances 0.000 description 2
- 239000008223 sterile water Substances 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 239000000273 veterinary drug Substances 0.000 description 2
- 241000282412 Homo Species 0.000 description 1
- 239000012880 LB liquid culture medium Substances 0.000 description 1
- 241000699666 Mus <mouse, genus> Species 0.000 description 1
- 230000009435 amidation Effects 0.000 description 1
- 238000007112 amidation reaction Methods 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 208000022531 anorexia Diseases 0.000 description 1
- 230000003385 bacteriostatic effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000003115 biocidal effect Effects 0.000 description 1
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 1
- 239000003054 catalyst Substances 0.000 description 1
- 238000004440 column chromatography Methods 0.000 description 1
- 230000008878 coupling Effects 0.000 description 1
- 238000005859 coupling reaction Methods 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 230000002354 daily effect Effects 0.000 description 1
- 206010061428 decreased appetite Diseases 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- XDNBJTQLKCIJBV-GOSISDBHSA-N diethoxy-[4-[(r)-methylsulfinyl]phenoxy]-sulfanylidene-$l^{5}-phosphane Chemical compound CCOP(=S)(OCC)OC1=CC=C([S@@](C)=O)C=C1 XDNBJTQLKCIJBV-GOSISDBHSA-N 0.000 description 1
- 238000003113 dilution method Methods 0.000 description 1
- 238000007598 dipping method Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000002224 dissection Methods 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 230000035622 drinking Effects 0.000 description 1
- 238000009509 drug development Methods 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 206010016256 fatigue Diseases 0.000 description 1
- 230000002550 fecal effect Effects 0.000 description 1
- 210000003608 fece Anatomy 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 238000004896 high resolution mass spectrometry Methods 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 238000007912 intraperitoneal administration Methods 0.000 description 1
- 244000144972 livestock Species 0.000 description 1
- 208000004396 mastitis Diseases 0.000 description 1
- 230000003340 mental effect Effects 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- 235000013336 milk Nutrition 0.000 description 1
- 239000008267 milk Substances 0.000 description 1
- 210000004080 milk Anatomy 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 230000010355 oscillation Effects 0.000 description 1
- 230000001575 pathological effect Effects 0.000 description 1
- 239000003761 preservation solution Substances 0.000 description 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000002390 rotary evaporation Methods 0.000 description 1
- 229920006395 saturated elastomer Polymers 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 235000015096 spirit Nutrition 0.000 description 1
- 238000000967 suction filtration Methods 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 230000000007 visual effect Effects 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Images
Classifications
-
- 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
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses a conjugate FST and a preparation method and application thereof. The invention relates to a method for preparing florfenicol succinate, (3H-1, 2, 3-triazolo [4, 5-B)]Pyridine-3-oxy) tri-1-pyrrolidinylhexafluorophosphate in solvent 1, in N 2 Stirring and reacting for 0.4-0.6 h at room temperature in the atmosphere to obtain reaction liquid 1; dissolving sulfathiazole with a solvent 2, adding the solution into the reaction solution 1, and adding the solution into the reaction solution N 2 And under the atmosphere, continuously adding N, N-diisopropylethylamine, reacting at room temperature for 5-7 h, stopping the reaction, removing the solvent, and purifying to obtain the conjugate FST. The preparation method is simple and high in yield, and the obtained conjugate FST has good sterilization and bacteriostasis capabilities on various bacteria, can be used as a broad-spectrum antibacterial drug, and effectively reduces pathogenic bacteria infection.
Description
Technical Field
The invention belongs to the technical field of antibiotic drug development, and particularly relates to a conjugate FST and a preparation method and application thereof.
Background
Antibiotics are the most effective drugs found in humans from soil and metabolites of various microorganisms to treat bacterial infections. With the abuse of antibiotics, bacteria begin to have drug resistance to the antibiotics and become drug-resistant bacteria, so that the difficulty of treating diseases of human and animals is improved. Although the developed new antibiotics can kill drug-resistant bacteria, the development of antibiotics requires screening in soil and various products of microorganisms therein, which requires high cost and long time, and the growth rate of bacterial drug resistance is far faster than the development rate of new antibiotics.
Disclosure of Invention
In order to solve the problems of the prior art, the primary object of the present invention is to provide a conjugate FST.
The invention further aims to provide a preparation method of the conjugate FST.
The invention also aims to provide the application of the conjugate FST in preparing medicines for resisting bacterial infection.
The invention is realized by the following steps that a conjugate FST has a molecular structural formula shown as the following formula (I):
the invention further discloses a pharmaceutically acceptable salt of the conjugate FST, which is selected from any one of salts formed by the conjugate FST and hydrochloric acid, sulfuric acid, phosphoric acid, acetic acid, fumaric acid, maleic acid, oxalic acid, malonic acid, glutamic acid, aspartic acid, succinic acid, citric acid and malic acid.
Preferably, the pharmaceutically acceptable salt of the conjugate FST is a compound with a molecular structural formula shown as the following formula (II):
the invention further discloses a preparation method of the conjugate FST, which comprises the following steps:
(1) Florfenicol succinate, (3H-1, 2, 3-triazolo [4, 5-B)]Pyridine-3-oxy) tri-1-pyrrolidinylhexafluorophosphate in solvent 1, in N 2 Stirring and reacting for 0.4-0.6 h at room temperature in an atmosphere to obtain a reaction solution 1;
(2) Dissolving sulfathiazole in solvent 2, adding into the reaction solution 1, and reacting under N 2 And under the atmosphere, continuously adding N, N-diisopropylethylamine, reacting at room temperature for 5-7 h, stopping the reaction, removing the solvent, and purifying to obtain the conjugate FST.
Preferably, in the step (1), the molar volume ratio of the florfenicol succinate, the (3H-1, 2, 3-triazolo [4,5-B ] pyridine-3-oxy) tri-1-pyrrolidinyl hexafluorophosphate and the solvent 1 is 0.65mmol:0.78mmol: 4-6 mL.
Preferably, in the step (2), the molar volume ratio of the sulfathiazole, the solvent 2, the solvent 1 and the N, N-diisopropylethylamine is 0.65mmol: 6-8 mL: 4-6 mL:1.3mmol.
Preferably, in the step (1) and the step (2), the solvent 1 and the solvent 2 are DMF.
Preferably, in the step (1), the florfenicol succinate is prepared by the following steps: adding florfenicol and succinic anhydride into acetone, carrying out reflux reaction for 5-7 h under the catalysis of 4-dimethylpyridine, and purifying a reaction product to obtain florfenicol succinate; wherein the molar ratio of the florfenicol to the succinic anhydride is 1.5:1, the 4-lutidine accounts for 4 percent of the mass of the florfenicol.
The invention further discloses application of the conjugate FST or pharmaceutically acceptable salts thereof in preparing medicaments for treating infectious diseases.
Preferably, the infectious diseases are infectious diseases caused by bacteria in human beings or animals, and the bacteria comprise escherichia coli and staphylococcus aureus.
The invention further discloses a medicament for treating infectious diseases, which comprises the conjugate FST or the pharmaceutically acceptable salt of the conjugate FST, and at least one pharmaceutically acceptable carrier, excipient or diluent of the conjugate FST or the pharmaceutically acceptable salt thereof
The invention overcomes the defects of the prior art and provides a conjugate FST as well as a preparation method and application thereof, the invention is based on a drug coupling method, a novel conjugate compound is formed by coupling the structure basis of the existing drug, and specifically, florfenicol succinate is obtained by the reflux reaction of florfenicol and succinic anhydride which are dissolved in acetone under the catalysis of 4-lutidine (DMAP), and then the FST is formed by amidation of the carboxyl end of the florfenicol succinate and the amino end of sulfathiazole. The synthetic route is shown as the following formula:
bacterial susceptibility experiments are carried out on the obtained conjugate FST, and the result shows that the conjugate FST has the activity of broad-spectrum antibiotics.
Compared with the defects and shortcomings of the prior art, the invention has the following beneficial effects:
(1) The conjugate FST has good sterilization and bacteriostasis capabilities on various bacteria, can be used as a broad-spectrum antibacterial drug, and effectively reduces pathogenic bacteria infection;
(2) The conjugate FST is simple in preparation method and high in yield.
Drawings
FIG. 1 is a high resolution mass spectrum of FST of a conjugate of the invention;
FIG. 2 shows the mortality of C57 mice treated with each drug group after APEC-O78 infection in application example 2 of the present invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and do not limit the invention.
Example 1
(1) Adding 1.5mmol of florfenicol and 1mmol of succinic anhydride into 20mL of acetone, adding 4wt% of 4-Dimethylpyridine (DMAP) of florfenicol (catalyst) into the system, refluxing for 6h, performing rotary evaporation on the reaction solution at 50 ℃ to remove the solvent, and then slowly adding saturated NaHCO at 50 DEG C 3 And (3) filtering the solution until no bubbles are generated in the solution, slowly adding 0.1mol of hydrochloric acid solution to the pH =2.0 after impurities are removed, refrigerating the liquid in a refrigerator at the temperature of 4 ℃, gradually separating out white solids, performing suction filtration, and washing with distilled water for several times to obtain the compound florfenicol succinate, wherein the yield is 72.7%.
Nuclear magnetic resonance hydrogen spectrum of the florfenicol succinate compound (A) 1 H-NMR) carbon spectrum (C) 13 C-NMR) and high resolution mass spectra HRMS data as follows:
1 H-NMR(400MHz，DMSO-d 6 )δ12.33(s，1H)，8.91(d，J＝8.8Hz，1H)，7.89(d，J＝8.5Hz，2H)，7.61(d，J＝8.4Hz，2H)，6.44(s，1H)，6.01(d，J＝4.3Hz，1H)，4.64-4.29(m，3H)，3.20(s，3H)，2.65(m，2H)，2.54(m，2H)。 13 C-NMR(101MHz，DMSO-d 6 )δ173.45，171.26，163.96，142.74，140.48，127.43，127.00，82.87，81.18，66.33，53.20，43.52，28.93，28.66。Chemical Formula：C 16 H 18 Cl 2 FNO 7 S，Exact Mass：457.0165，HRMS(+TOF MS)：475.0517[M+NH 4 ] + 。
(2) The compound florfenicol succinate (0.65 mmol) and (3H-1, 2, 3-triazolo [4, 5-B)]Pyridine-3-oxy) tris-1-pyrrolidinylhexafluorophosphate (Pyaop, 0.78 mmol) was added to bottle A, 5mL DMF was added, and then the mixture was passed through N 2 Protecting, and stirring for 0.5h at room temperature;
(3) Sulfathiazole (0.65 mmol) was weighed into bottle B, dissolved in 5mL of DMF, transferred to bottle A, washed with 2mL of DMF and transferred to bottle A, N 2 After protection, N-diisopropylethylamine (DIPEA, 1.3 mmol) was added and the reaction was carried out at room temperature for 6 hours. After the reaction was terminated, the solvent was lyophilized with a freeze dryer, and then column chromatography (developing solvent ratio dichloromethane/methanol =20 = 1) was performed to obtain the conjugate FST with a yield of 45% to 55%.
The High Resolution Mass Spectrum (HRMS) of the FST of the conjugate is shown in figure 1, its nuclear magnetic resonance hydrogen spectrum ( 1 H-NMR), carbon spectrum ( 13 C-NMR) and High Resolution Mass Spectrometry (HRMS) data as follows:
1 H NMR(400MHz,DMSO-d 6 )δ12.68(s,1H),10.34(s,1H),8.91(d,J＝8.8Hz,1H),7.92–7.85(m,2H),7.71(q,J＝8.7Hz,4H),7.62(d,J＝8.2Hz,2H),7.24(d,J＝4.6Hz,1H),6.80(d,J＝4.6Hz,1H),6.44(s,1H),6.01(d,J＝4.4Hz,1H),4.66–4.28(m,3H),3.18(s,3H),2.79–2.63(m,4H). 13 C NMR(101MHz,DMSO-d 6 )δ171.51,170.41,168.80,163.99,142.76,142.32,140.52,136.29,127.50,127.08,127.05,124.47,118.52,108.16,82.92,72.65,66.35,53.23,43.52,30.90,28.67.
Chemical Formula：C 25 H 25 Cl 2 FN 4 O 8 S 3 ，Exact Mass：694.0196，HRMS(+TOF MS)：695.0260[M+H] + 。
application example FST susceptibility test for various bacteria
1. Experimental equipment
The device comprises a 96-well plate, a 5 uL-50 uL micropipettor, a 100 uL-1000 uL micropipettor, a matched gun head, a 15mL centrifugal tube, a My turbidimetric tube, an ultra-clean workbench, an autoclave, a blast drying oven, a biochemical incubator, an MH broth culture medium and sterile water.
Relating to strains and sources: escherichia coli 25922 and staphylococcus aureus 29213 are purchased from ATCC, escherichia coli 278, escherichia coli 892, escherichia coli 257, escherichia coli 783, staphylococcus aureus 364 and staphylococcus aureus 424 are all separated from mammitis milk samples, belong to clinical isolates and are deposited in the research institute of Lanzhou livestock and veterinary drugs, national academy of agricultural sciences.
2. Experimental method
1. Placing the prepared MH culture medium, sterile water, a matched gun head and a centrifuge tube into an autoclave for sterilization, wherein the sterilization parameter is 121 ℃, and the sterilization time is 25min.
2. Taking 15mL of a centrifuge tube, respectively and sequentially adding 7mL of nutrient broth and 10 mu L of bacteria liquid, and incubating for 18-24 h at 37 ℃ in a constant-temperature oscillator at an oscillation speed of 170-180 r/min to obtain resuscitation bacteria liquid.
3. The method is characterized in that a micro-double dilution method is adopted for determination, a sterile 96-well plate is taken, 200 mu L of antibacterial drugs are added into a first hole, 100L of MH broth culture medium is respectively added into a second hole to a tenth hole, 100 mu L of MH broth culture medium is sucked from the first hole and added into the second hole, the MH broth culture medium and the MH broth culture medium are mixed uniformly, 100 mu L of MH broth culture medium is sucked from the first hole and then sucked into the third hole, the process is repeated, and 100 mu L of MH broth culture medium is sucked from the tenth hole and then discarded. The drug concentrations are in turn: 200. 100, 50, 25, 12.5, 6.25, 3.125, 1.56, 0.78, 0.39. Mu. Mol/L, 200. Mu.L of the bacterial suspension was added to the eleventh well, and 200. Mu. LMH medium was added to the twelfth well. Then 100. Mu.L of the bacterial suspension was added to each of the 1 to 10 wells so that the final concentration of the bacterial suspension in each tube was about 5X 10 5 CFU/ML. And (3) placing the inoculated 96-well plate in an incubator at 37 ℃ for culture, taking out the 96-well plate for 24h, visually observing the clarity in the hole, and taking the concentration corresponding to the clarified and precipitate-free final hole in the hole by visual observation as the MIC value of the medicine.
3. Results of the experiment
The results of the experiment are shown in table 1 below:
TABLE 1 MIC values of FST against different strains
| Bacterial strains | MIC(μmol/L) |
| Escherichia coli 25922 | 6.25 |
| Staphylococcus aureus 29213 | 6.25 |
| Escherichia coli 278 | 3.12 |
| Escherichia coli 892 | 6.25 |
| Escherichia coli 257 | 6.25 |
| Escherichia coli 783 | 1.56 |
| Staphylococcus aureus 364 | 3.12 |
| Staphylococcus aureus 424 | 6.25 |
As can be seen from Table 1, the FST of the invention has good bacteriostatic effect on the strains used in the test.
Application example 2 evaluation of FST in vivo drug efficacy
1. Experimental methods
1. Preparation and dilution of test bacterium liquid
Firstly, APEC-O78 (Escherichia coli, CVCC1490, available from China institute for veterinary drug) preservation solution is taken out from a refrigerator at the temperature of-80 ℃, and the following operations are carried out:
(1) Dipping a small amount of bacterial liquid by using a sterile inoculating loop, scribing on an LB agar culture medium, and inversely placing a flat plate in a constant-temperature incubator at 37 ℃ for 14 hours;
(2) A single colony on an LB plate is picked by an inoculating loop and inoculated in 200mL of LB liquid culture medium, and the single colony is placed in a constant temperature shaking incubator at 37 ℃ for culture for 14h for later use;
(3) Centrifuging at 4000rpm at 4 deg.C for 5min, re-suspending with sterile PBS, sucking 100 μ L of bacteria culture solution, adding 900 μ L of sterile PBS, shaking, and mixing to obtain 10 -1 Diluting the solution;
(4) Get 10 -1 Diluting the diluent to 10 times -7 、10 -8 And 10 -9 And (3) waiting for a series of bacterial suspensions, diluting each bacterial suspension for 3 times, inoculating each bacterial suspension onto an LB agar plate, culturing for 14 hours, counting bacteria, and multiplying the bacterial suspensions by the dilution times to obtain the total number of colonies.
2. Poison-counteracting dose screening
15C 57 mice were randomly divided into 3 groups, and the prepared bacterial culture was inoculated by intraperitoneal injection, each group was administered with 0.1 mL/mouse. Observing for 4 days, recording the death condition of the mice, and determining the toxic counteracting dose.
3. Test grouping and administration
Mice were randomized into 8 groups (n = 8) before the start of the experiment: a control group, an infection model group, a florfenicol FF group (20 mg/kg), a sulfathiazole ST group (14.3 mg/kg), an FF + ST group (20 mg/kg +14.3 mg/kg), an FST-L group (19.45 mg/kg), an FST-M group (38.9 mg/kg), and an FST-H group (77.8 mg/kg). Healthy controls were injected i.p. with the same volume of sterile PBS solution. According to the measured toxicity attacking dose, the prepared bacterial liquid is administered to the mice except the healthy control group in an intraperitoneal injection mode, the medicine is administered once in the intraperitoneal injection mode every day after infection, and the healthy control group is perfused with the medicine solvent with the same volume. Treatment continued for 4d. After 4d, the mice were all sacrificed and tissues were collected for analysis.
4. Observation index
Clinical symptoms are as follows: during the whole experiment period, the clinical conditions of feeding, drinking, diarrhea, furs, standing, spirits and the like of each group of mice are observed every day; the dead mice were subjected to pathological dissection and the death was recorded.
The mortality rate was calculated: during the trial, the number of morbidity and mortality of each group of mice was observed and recorded in detail daily, and the mortality of each group of mice was calculated:
mortality = number of dead animals/number of test animals
2. Test results
The results of the experiments are shown in FIG. 2, which is the mortality rate of C57 mice treated with each drug group after APEC-O78 infection.
As can be seen from figure 2, the model group died at the beginning of 12 hours, and all died after 24-48 hours reached the peak, and mice were lassitude, anorexia, vertical hair, and loose water in the whole process. The FF group in the control group died 3 mice within 12-98 h, but no death occurred in the following days, and the mice had better spirit, good appetite and reduced diarrhea, and were basically normal on day 5. The ST group died at 12h, reached peak death at 24-48 h, and died continuously for several days until all died on day 5. 4 mice in the equimolar FF and ST groups died between 24 and 96 hours until no death occurred in 96 hours, and the spirit and appetite gradually recovered to normal and the diarrhea decreased. The low and medium dose FST groups had 3 and 4 deaths each between 24 and 96 hours, and recovery began after 96 hours, with no subsequent deaths, mental, appetite, and fecal normality. 2 deaths occurred in the high dose FST group between 24 and 96 hours, and then no deaths occurred, and mice were normal in spirit, appetite, and feces. The blank group did not die throughout the experiment.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents and improvements made within the spirit and principle of the present invention are intended to be included within the scope of the present invention.
Claims (9)
1. A conjugate FST, wherein the molecular structural formula of the conjugate FST is shown as the following formula (I):
2. the pharmaceutically acceptable salt of the conjugate FST according to claim 1, wherein the pharmaceutically acceptable salt of the conjugate FST is selected from any one of salts of the conjugate FST with hydrochloric acid, sulfuric acid, phosphoric acid, acetic acid, fumaric acid, maleic acid, oxalic acid, malonic acid, glutamic acid, aspartic acid, succinic acid, citric acid and malic acid.
3. The pharmaceutically acceptable salt of claim 2, wherein the pharmaceutically acceptable salt of the conjugate FST is a compound having a molecular structural formula of formula (II):
4. a method of preparing the conjugate FST of claim 1, comprising the steps of:
(1) Florfenicol succinate, (3H-1, 2, 3-triazolo [4, 5-B)]Pyridine-3-oxy) tri-1-pyrrolidinylhexafluorophosphate in solvent 1, in N 2 Stirring and reacting for 0.4-0.6 h at room temperature in an atmosphere to obtain a reaction solution 1;
(2) Dissolving sulfathiazole in solvent 2, adding into the reaction solution 1, and reacting under N 2 And under the atmosphere, continuously adding N, N-diisopropylethylamine, reacting at room temperature for 5-7 h, stopping the reaction, removing the solvent, and purifying to obtain the conjugate FST.
5. The preparation method according to claim 4, wherein in step (1), the molar volume ratio of the florfenicol succinate, (3H-1, 2, 3-triazolo [4,5-B ] pyridin-3-oxy) tri-1-pyrrolidinylhexafluorophosphate, solvent 1 is 0.65mmol:0.78mmol: 4-6 mL;
in the step (2), the molar volume ratio of the sulfathiazole, the solvent 2, the solvent 1 and the N, N-diisopropylethylamine is 0.65mmol: 6-8 mL: 4-6 mL:1.3mmol;
in the step (1) and the step (2), the solvent 1 and the solvent 2 are DMF.
6. The method of claim 4, wherein in step (1), the florfenicol succinate is prepared by: adding florfenicol and succinic anhydride into acetone, carrying out reflux reaction for 5-7 h under the catalysis of 4-dimethylpyridine, and purifying a reaction product to obtain florfenicol succinate; wherein the molar ratio of the florfenicol to the succinic anhydride is 1.5:1, the mass of the 4-lutidine is 4 percent of that of the florfenicol.
7. Use of the conjugate FST according to claim 1 or the pharmaceutically acceptable salt according to claim 2 for the manufacture of a medicament for the treatment of an infectious disease.
8. The use of claim 7, wherein the infectious disease is caused by bacteria including Escherichia coli and Staphylococcus aureus in a human or animal.
9. A medicament for the treatment of an infectious disease, which medicament comprises the conjugate FST according to claim 1 or a pharmaceutically acceptable salt of the conjugate FST according to claim 2 or claim 3, and at least one of the conjugate FST or a pharmaceutically acceptable salt of the conjugate FST in a pharmaceutically acceptable carrier, excipient or diluent.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211415973.2A CN115745909B (en) | 2022-11-12 | 2022-11-12 | Conjugate FST and preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211415973.2A CN115745909B (en) | 2022-11-12 | 2022-11-12 | Conjugate FST and preparation method and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN115745909A true CN115745909A (en) | 2023-03-07 |
| CN115745909B CN115745909B (en) | 2024-01-26 |
Family
ID=85370299
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202211415973.2A Active CN115745909B (en) | 2022-11-12 | 2022-11-12 | Conjugate FST and preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN115745909B (en) |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101289416A (en) * | 2008-03-03 | 2008-10-22 | 西南大学 | High-water-solubility florfenicol prodrug quickly released in vivo |
| CN102151269A (en) * | 2011-02-21 | 2011-08-17 | 天津市海纳德动物药业有限公司 | Composition for treating pig paratyphus and preparation method thereof |
| CN104003917A (en) * | 2014-03-14 | 2014-08-27 | 河北润普兽药有限公司 | Preparing method of florfenicol sodium succinate microcrystals |
| CN110642764A (en) * | 2018-06-27 | 2020-01-03 | 黄焕军 | Preparation method of water-soluble florfenicol sodium salt |
| CN113975404A (en) * | 2021-09-22 | 2022-01-28 | 中国农业科学院兰州畜牧与兽药研究所 | Florfenicol polypeptide derivative and application thereof |
| CN115286544A (en) * | 2022-10-08 | 2022-11-04 | 世华合创生物技术开发(山东)有限公司 | Florfenicol derivative, preparation method and application thereof in resisting bacterial infection |
-
2022
- 2022-11-12 CN CN202211415973.2A patent/CN115745909B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101289416A (en) * | 2008-03-03 | 2008-10-22 | 西南大学 | High-water-solubility florfenicol prodrug quickly released in vivo |
| CN102151269A (en) * | 2011-02-21 | 2011-08-17 | 天津市海纳德动物药业有限公司 | Composition for treating pig paratyphus and preparation method thereof |
| CN104003917A (en) * | 2014-03-14 | 2014-08-27 | 河北润普兽药有限公司 | Preparing method of florfenicol sodium succinate microcrystals |
| CN110642764A (en) * | 2018-06-27 | 2020-01-03 | 黄焕军 | Preparation method of water-soluble florfenicol sodium salt |
| CN113975404A (en) * | 2021-09-22 | 2022-01-28 | 中国农业科学院兰州畜牧与兽药研究所 | Florfenicol polypeptide derivative and application thereof |
| CN115286544A (en) * | 2022-10-08 | 2022-11-04 | 世华合创生物技术开发(山东)有限公司 | Florfenicol derivative, preparation method and application thereof in resisting bacterial infection |
Also Published As
| Publication number | Publication date |
|---|---|
| CN115745909B (en) | 2024-01-26 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN113149929B (en) | Pleuromutilin derivative with 1,3, 4-oxadiazole side chain and preparation and application thereof | |
| CN114230519B (en) | Pleuromutilin cinnamate compounds with drug-resistant bacteria resisting activity, and synthetic method and application thereof | |
| JPS60237099A (en) | Antibiotic m43a, m43b, m43c and m43d and related compounds | |
| DK145381B (en) | PROCEDURE FOR THE PREPARATION OF AN ANTIBIOTIC SUBSTANCE, CALLED N-ACETYLDEHYDROTHIENAMYCINE, OR PHARMACEUTICAL ACCEPTABLE SALTS THEREOF | |
| CN111499628A (en) | 2-furoylamide compound and preparation and application thereof | |
| CN114634498A (en) | Pleuromutilin derivative containing thiazole-pyridine benzyl quaternary ammonium salt side chain as well as preparation method and application thereof | |
| CN115745909A (en) | Conjugate FST and preparation method and application thereof | |
| JPS60231698A (en) | Novel glycopeptide antibiotic and manufacture | |
| JPS5850999B2 (en) | Houteimycin AE and its manufacturing method | |
| JPH0631301B2 (en) | Antibacterial 9-deoxo-9a-allyl and propargyl-9a-aza-9a-homoerythromycin A derivatives | |
| CN103421100B (en) | A kind of antibacterial peptide and its preparation method and application | |
| WO2023115691A1 (en) | Pleuromutilin derivative with amino side chain, and preparation method therefor and use thereof | |
| CN103232447B (en) | 3-acetyl-5-acetylimino-2-(N-phenothiazinyl)-1,3,4-thiadiazole, and preparation method and application thereof | |
| CN114940671A (en) | Pleuromutilin derivative with 4-aminobenzenethiol side chain as well as preparation method and application thereof | |
| CN107382893B (en) | Linezolid base cation amphiphilic compound with antibacterial activity and preparation method thereof | |
| CN113975404A (en) | Florfenicol polypeptide derivative and application thereof | |
| FI101402B (en) | A method for preparing a new antibiotic, balhimycin | |
| CN104892507B (en) | Water-soluble methanesulfonic acid sarafloxacin and preparation method thereof | |
| JPS60141293A (en) | Novel carcinostatic antibiotic substance 81-484 and its production | |
| JP4541405B2 (en) | Antibiotic compound | |
| CN107417601A (en) | The aryl-pyridine of 2 acid amides sulfonyl of substitution 4,6 with antibacterial activity | |
| CN117903255B (en) | Antibacterial peptide and preparation method and application thereof | |
| JPS60133000A (en) | Glycopeptide biologically converted product | |
| CN113912484B (en) | 1,4, 6-trihydroxy-8-branched-9, 10-anthraquinone compound and application thereof in preparation of bacteriostat | |
| CN117417280A (en) | Pleuromutilin derivative containing cycloalkyl, preparation method and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |