CN108658991B - Preparation method and application of 3, 5-disubstituted methylpyrazolo [1,5-a ] pyrimidine-7-phenolate analogue and derivative - Google Patents

Preparation method and application of 3, 5-disubstituted methylpyrazolo [1,5-a ] pyrimidine-7-phenolate analogue and derivative Download PDF

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CN108658991B
CN108658991B CN201710212683.0A CN201710212683A CN108658991B CN 108658991 B CN108658991 B CN 108658991B CN 201710212683 A CN201710212683 A CN 201710212683A CN 108658991 B CN108658991 B CN 108658991B
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徐利锋
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Abstract

The invention aims to provide 3- (2, 4-difluorophenyl) -5-substituted-2-methylpyrazolo [1,5-a ]]Pyrimidine-7-phenolate derivatives and analogues have the following general formula or prodrug, and provide a preparation method, a pharmacological activity experimental method and pharmacological activity thereof, which are used for discovering antibacterial and antifungal activity, pharmaceutical chemistry research and a preparation method, and application of the compounds as antibacterial and antifungal medicines.R and M in the structural formula I + The definition is shown in the specification. The structural formula I of the invention is 3- (2, 4-difluorophenyl) -5-substituted-2-methylpyrazolo [1,5-a ]]Pyrimidine-7-phenolate derivatives and analogs, including use in combination with other known antibacterial and antifungal agents, including use in combination with therapeutic agents that accompany various complications of bacterial infections, such as inflammation, viruses, and immune system disorders.

Description

Preparation method and application of 3, 5-disubstituted methylpyrazolo [1,5-a ] pyrimidine-7-phenolate analogue and derivative
Technical Field
The present invention relates to the discovery of 3- (2, 4-difluorophenyl) -5-substituted-2-methylpyrazolo [1,5-a ] pyrimidine-7-phenoxide analogues and derivatives as antibacterial and antifungal activity, pharmaceutical chemistry studies and methods of preparation. The invention also relates to application of the compounds as antibacterial and antifungal medicaments.
Background
Due to the large number of antibacterial drugs of abuse, the available antibacterial drugs may develop resistant strains, exhibit resistance to various antibacterial drugs, such as methicillin-resistant staphylococcus aureus (MRSA), vancomycin-resistant enterococci (VRE), etc., known as "superbacteria" are produced. The spread of infection "superbacteria" has constituted a serious threat to human health, and thus it has been urgent to develop new antibacterial agents active against drug-resistant bacteria.
The entire literature is searched, and no report on the use of 3- (2, 4-difluorophenyl) -5-substituted-2-methylpyrazolo [1,5-a ] pyrimidine-7-phenoxide with antibacterial and antifungal activity is found. Other biological activity research patents of compounds with different structures and similarity in structural parent nucleus are: WO 2008062026, EP 2086947 and US 20130252951, which patent invent compounds which differ from the compounds of the invention in structure, but which have similarities in the parent nucleus structure, are also independent of the invention in their biological activity and application indications, and are all useful in anti-inflammatory therapy; the compounds of US 20160022742 differ from the compounds of the invention only in that they have similarities in the parent nucleus structure and their biological activity and use are not relevant to the present invention and are useful in the treatment of liver dysfunction.
In summary, all the documents are searched so far, and the patent invention and articles of the application of the compound structure 3- (2, 4-difluorophenyl) -5-substituted-2-methylpyrazolo [1,5-a ] pyrimidine-7-phenolate derivatives and analogues represented by the structural formula I and the antibacterial and antifungal bioactivity of the compound are not reported.
Disclosure of Invention
The invention aims to provide chemical synthesis and preparation of 3- (2, 4-difluorophenyl) -5-substituted-2-methylpyrazolo [1,5-a ] pyrimidine-7-phenolate derivatives, which have the following general formula and prodrugs, and provide preparation, pharmacological activity experimental methods and pharmacological activity application thereof.
The invention aims at realizing the following molecular formula structure:
wherein M is described by structural formula I + The method comprises the following steps: li (Li) + 、K + 、Na + 、Ca + /2, Mg + /2, Cu + 2 and Fe + One of the compounds/2 may be a salt formed by reacting heterocyclic phenolic hydroxyl groups with various bases and alkali metals, for example, inorganic bases such as metal Li, metal Na, metal K, sodium carbonate, sodium hydride, sodium hydroxide, potassium hydroxide and the like, organic bases to form phenoxide or double salts; the nitrogen atom in the structural formula I has basic passable acid,such as hydrochloric acid, sulfuric acid, nitric acid, fumaric acid, maleic acid, succinic acid, acetic acid, citric acid, tartaric acid, carbonic acid, phosphoric acid, oxalic acid, and the like, to form inorganic acid salts, organic acid salts, or double salts.
Wherein, the 3- (2, 4-difluorophenyl) -5-substituted-2-methylpyrazolo [1,5-a ] pyrimidine-7-phenoxide derivatives and analogues as shown in the structural formula I, R as shown in the structural formula I is H, optionally substituted aryl, alicyclic and/or fused combination of aryl and alicyclic, and can be one of substituent groups 1, 2, 3, 4, 5, 6, 7, 8 and 9 as shown in the structural formula II, wherein, the alicyclic ring of the aromatic rings as shown in the chemical structures 8 and 9 is a three-carbon, four-carbon or five-carbon alicyclic ring.
Wherein X depicted in formula II may be absent, may each be the same or different substituent, and is optionally substituted hydrogen, halogen, hydroxy, mercapto, cyano, carbonyl, substituted carbonyl, aldehyde, ketone, nitro, carboxyl, substituted carboxyl, carboxylate, secondary amino, tertiary amino, di-, tri-, tetra-, penta-, or hexa-alkyl oxy, arylalkoxy, aryloxy, heteroaryloxy, alkylthio, mercapto, arylalkylthio, arylthio, heteroarylthio, ester, acyloxy, phosphoric acid oxy, sulfonic acid oxy, aryloxy, quaternary ammonium salt, amide, hydrazine, oxime, hydrazone, nitrogenous aliphatic hydrocarbon, nitrogenous aromatic hydrocarbon, nitrogenous cyclic, nitrogenous aromatic heterocyclic, phosphide, phosphate, mono-, di-, tri-, tetra-, or pentacarbon alkyl, and combinations thereof.
The 3- (2, 4-difluorophenyl) -5-substituted-2-methylpyrazolo [1,5-a ] pyrimidine-7-phenolate derivatives and analogues as shown in the structural formula I comprise antibacterial pharmacological activity and application as antibacterial drugs, antifungal pharmacological activity and application as antifungal drugs, wherein the antifungal pharmacological activity and application as antifungal drugs comprise compatibility with other known antibacterial, antifungal, anti-inflammatory and antiviral drugs, and immune drugs, and compatibility with treatment drugs for inflammation and inflammation diseases, fungi and fungal diseases, viruses and viral diseases, immune system diseases and other complications associated with bacterial infection, and the like, and the dosage of the 3- (2, 4-difluorophenyl) -5-substituted-2-methylpyrazolo [1,5-a ] pyrimidine-7-phenolate derivatives and analogues is 0.02 mg/kg-250 mg/kg (intravenous, intramuscular injection, oral, local administration and other administration routes); various methods of treatment and pathway treatments, wherein the bacteria are gram positive bacteria: staphylococci, pneumococci, enterococci, streptococci, streptococcus bovis, streptococcus pneumoniae, streptococcus mutans, streptococcus pyogenes, streptococcus agalactiae, streptococcus viridae, streptococcus bovis, streptococcus agalactiae B, group streptococcus viridae, diphtheria bacillus, clostridium, erysipelas, bacillus anthracis, bacillus tetani, bacillus cereus, bacillus subtilis, clostridium, bacillus cereus, bacillus subtilis, bacillus anthracis, bacillus diphtheria, clostridium, tetani, bacillus perfringens, clostridium perfringens spiral, actinomycetes, tubercle bacteria, gram positive drug-resistant bacteria, methicillin-resistant bacteria Lin Putao, vancomycin-resistant bacteria, staphylococcus-induced clindamycin-resistant bacteria, high-level aminoglycosides bacteria, penicillin-resistant streptococcus pneumoniae, multi-drug-resistant bacillus baumannii, drug-resistant and multi-resistant bacteria, bacillus tuberculosis, streptococcus, enterococcus, pseudomonas aeruginosa, escherichia coli, pseudomonas, neisseria, and other bacteria resistant to influenza, and the bacteria of the families of the group of the bacteria.
The specific compound structures of the 3- (2, 4-difluorophenyl) -5-substituted-2-methylpyrazolo [1,5-a ] pyrimidine-7-phenolate derivatives and analogues of the structural formula I are shown in Table 1, and examples 1 to 33, but are not limited to the examples: 3- (2, 4-difluorophenyl) -2-methyl-5- (4- (trifluoromethyl) phenyl) pyrazolo [1,5-a ] pyrimidin-7-ol sodium, 5- (4-cyanophenyl) -3- (2, 4-difluorophenyl) -2-methylpyrazolo [1,5-a ] pyrimidin-7-ol sodium, 3- (2, 4-difluorophenyl) -2-methyl-5- (4-nitrophenyl) pyrazolo [1,5-a ] pyrimidin-7-ol sodium, 3- (2, 4-difluorophenyl) -5- (4-methoxyphenyl) -2-methylpyrazolo [1,5-a ] pyrimidin-7-ol sodium, 3- (2, 4-difluorophenyl) -5- (4- (dimethylamino) phenyl) -2-methylpyrazolo [1,5-a ] pyrimidin-7-ol sodium, 3- (2, 4-difluorophenyl) -2-methyl-5- (4-phenol sodium) -pyrazolo [1,5-a ] pyrimidine-7-ol sodium, 5- (4-nitrophenyl) -2-methylpyrazolo [1,5-a ] pyrimidine-7-ol sodium, 5- (4-methoxyphenyl) -2-methylpyrazolo [1,5-a ] pyrimidine-7-ol sodium, 3- (4-fluorophenyl) -2-methyl-pyrazolo [1,5-a ] pyrimidine-7-ol sodium, 1-b-4-carboxylate sodium, sodium 5- (4-sulfoacid sodium phenyl) -3- (2, 4-difluorophenyl) -2-methylpyrazolo [1,5-a ] pyrimidin-7-ol, sodium 3- (2, 4-difluorophenyl) -2-methyl-5- (4-sulfamoylphenyl) pyrazolo [1,5-a ] pyrimidin-7-ol, sodium 3- (2, 4-difluorophenyl) -2-methyl-5- (4-phenoxyphenyl) pyrazolo [1,5-a ] pyrimidin-7-ol, sodium 5- (4-formylphenyl) -3- (2, 4-difluorophenyl) -2-methylpyrazolo [1,5-a ] pyrimidin-7-ol, sodium 3, 5-bis (2, 4-difluorophenyl) -2-methylpyrazolo [1,5-a ] pyrimidin-7-ol, sodium 5- (4-ethoxyphenyl) -3- (2, 4-difluorophenyl) -2-methylpyrazolo [1,5-a ] pyrimidin-7-ol, sodium 3- (2, 4-difluorophenyl) -2-methyl-pyrazolo [1,5-a ] pyrazolo [ 7-ol, sodium 3- (2, 4-difluorophenyl) -2-methylpyrazolo [1,5-a ] pyrimidine-7-ol, sodium 3- (3-methyl-7-o-7-ol, sodium 5- (6-cyanopyridin-3-yl) -3- (2, 4-difluorophenyl) -2-methylpyrazolo [1,5-a ] pyrimidin-7-carboxylate, sodium 3- (2, 4-difluorophenyl) -2-methyl-5- (6-nitropyridin-3-yl) pyrazolo [1,5-a ] pyrimidin-7-carboxylate, sodium 3- (2, 4-difluorophenyl) -5- (6-methoxypyridin-3-yl) -2-methylpyrazolo [1,5-a ] pyrimidin-7-carboxylate, sodium 3- (2, 4-difluorophenyl) -5- (6- (dimethylamino) pyridin-3-yl) -2-methylpyrazolo [1,5-a ] pyrimidin-7-carboxylate, sodium 3- (2, 4-difluorophenyl) -5- (6-phenolate sodium pyridin-3-yl) -2-methylpyrazolo [1,5-a ] pyrimidin-7-carboxylate, sodium 5- (4-cyanophenyl) -3- (2, 4-difluorophenyl) -2-methylpyrazolo [1,5-a ] pyrimidin-7-carboxylate, potassium 3- (2, 4-difluorophenyl) -2-methylpyrazolo [1,5-a ] pyrimidin-7-carboxylate, sodium 3- (2, 4-difluorophenyl) -2-methylpyrazolo [ 1-3-yl ] pyrimidine-7-carboxylate, sodium, 3- (2, 4-difluorophenyl) -2-methyl-5- (4- (trifluoromethyl) phenyl) pyrazolo [1,5-a ] pyrimidin-7-phenol potassium, 3- (2, 4-difluorophenyl) -2-methyl-5- (4-nitrophenyl) pyrazolo [1,5-a ] pyrimidin-7-phenol potassium, 3- (2, 4-difluorophenyl) -5- (4- (dimethylamino) phenyl) -2-methylpyrazolo [1,5-a ] pyrimidin-7-phenol potassium, 3- (2, 4-difluorophenyl) -2-methyl-5- (4-sulfamoylphenyl) pyrazolo [1,5-a ] pyrimidin-7-phenol potassium, 3- (2, 4-difluorophenyl) -2-methyl-5- (6- (trifluoromethyl) pyridin-3-yl) pyrazolo [1,5-a ] pyrimidin-7-phenol potassium, 5- (6-cyanopyridin-3-yl) -3- (2, 4-difluorophenyl) -2-methylpyrazolo [1,5-a ] pyrimidine-7-phenol potassium, 3- (2, 4-difluorophenyl) -2-methyl-5-a ] pyrazolo [1,5-a ] pyrimidine-7-phenol sodium, 3-fluoro-1, 4-difluoro-a ] pyrazolo [1, 5-methyl-5- (6-trifluoromethyl) pyridin-3-yl), sodium 3- (2, 4-difluorophenyl) -2-methyl-5- (6-sulfamylpyridin-3-yl) pyrazolo [1,5-a ] pyrimidin-7-carboxylate, sodium 3- (2, 4-difluorophenyl) -2-methyl-5- (6-phenoxypyridin-3-yl) pyrazolo [1,5-a ] pyrimidin-7-carboxylate, sodium 3- (2, 4-difluorophenyl) -5- (4- ((2- (dimethylamino) ethyl) carbamoyl) phenyl) -2-methylpyrazolo [1,5-a ] pyrimidin-7-carboxylate, sodium 5- (6-formylpyridin-3-yl) -3- (2, 4-difluorophenyl) -2-methylpyrazolo [1,5-a ] pyrimidin-7-carboxylate, sodium 5- (6-acetoxypyridin-3-yl) -3- (2, 4-difluorophenyl) -2-methylpyrazolo [1,5-a ] pyrimidine-7-carboxylate.
Pharmacological activity of 3- (2, 4-difluorophenyl) -5-substituted-2-methylpyrazolo [1,5-a ] pyrimidine-7-phenoxide derivatives and analogues of formula I and use as antibacterial and antifungal agents, the various infections caused by bacterial and fungal infections also include inflammatory and inflammatory diseases associated with bacterial infections, fungal and fungal diseases, viral and viral diseases, and complications of immune system diseases: upper and lower respiratory tract infections, skin soft tissue infections, urinary tract infections, sepsis, endocarditis, etc. caused by methicillin-sensitive staphylococci, hemolytic streptococcus and pneumococcus; can also be used for urinary tract infection and pneumonia caused by haemophilus influenzae, proteus mirabilis, escherichia coli sensitive strain, gram positive coccus such as streptococcus and Streptococcus pneumoniae, and respiratory tract infection, urinary tract infection, skin soft tissue infection, septicemia, bone, joint infection, abdominal cavity infection and pelvic cavity infection caused by sensitive strain in haemophilus influenzae, escherichia coli and Proteus mirabilis, and infection such as haemolytic streptococcus, pneumococcus and sensitive staphylococcus aureus; endocarditis, gas gangrene, anaerobe infection, anthrax, syphilis, gonorrhea and the like caused by streptococcus grass and enterococcus.
3- (2, 4-difluorophenyl) -2-methylpyrazolo [1,5-a ] pyrimidine-7-phenoxide derivatives and analogues of formula I and their use, wherein the compounds are compatible or co-administered with at least one or a combination of known antibacterial, antifungal, anti-inflammatory agents or pharmaceutically acceptable salts or prodrugs of the agents, including but not limited to: beta-lactams: penicillin, procaine penicillin, benzathine penicillin, methicillin, oxacillin, cloxacillin, dicloxacillin sodium, ampicillin, amoxicillin, seatoxillin, carboxicillin, sulbenicillin, temoxicillin, furbenicillin, piperacillin, azlocillin, mezlocillin, ticarcillin, mexillin, apaxillin, apoxillin, ampicillin, flucloxacillin, sultaxillin, pivoxillin, phthalamicillin, bazacillin, bixacillin, furacillin, ceftriaxone, cefpirome, cefuroxime, ceftioxime, ceftiofur, cefathiamidine, cefacetonitrile, cefpiramide, cefazolin, fluxillin, flucloxillin, sulbactam cefmenoxime, cefoperazone, cefaclor, ceftizoxime, ceftazidime, cefonicid, cefdinir, cefixime, cefbuperazone, cefpiramide, ceftizoxime, cefpodoxime proxetil, ceftibetan, cefetamet, ceftizoxime, cefprozil, ceftibuzene, cefepime, cefalexin, cefradine, cefaclor, cefadroxil, cefamandole, cefsulodin, cefmetazole, cefotetan, cefminox, cefcapene, ceftazidime, ceftizoxime, cefradine, ceftriaxone, chlorocarbon, flomoxef, macrolides: dirithromycin, roxithromycin, clarithromycin, fluoroerythromycin, azithromycin, natamycin, tacmesilate, erythromycin, etomycin, clarithromycin, kitasamycin, milamycin, leucomycin, midecamycin, azithromycin, tsunamycin, spiramycin, acetylspiramycin, aminoglycosides: netilmicin, astemicin, arbekacin, ipramicin streptomycin, calicheamicin, gentamicin, tobramycin, amikacin, sisomicin, neomycin, paromomycin, fosamicin, minocycline, ipramicin, dbimicin, dardimicin, spectinomycin, streptomycin, kanamycin, etimicin, dbecaxing, amid alcohols: chloramphenicol, amber chloramphenicol, palmitomycetin, thiamphenicol, lincomycin, clindamycin phosphate, polypeptide polyenes: cyclosporin, teicoplanin, perlomycin polymyxin, vancomycin, norvancomycin, bacitracin, polymyxin B, fusidic acid, mi Kamei, rifamycins: rifabutin, rifapentine, rifaximin, rifampin, rifamycin, rifadine, quinolones: enoxacin, tolofloxacin, norfloxacin, ciprofloxacin, lomefloxacin, sefloxacin, pefloxacin, fleroxacin, temafloxacin, sarafloxacin, moxifloxacin, tervafloxacin, glafloxacin, ofloxacin, levofloxacin, pa Chu Shaxing, rufloxacin, sulfaisoxazole, sulfamethoxazole, sulfadiazine sodium, silver sulfadiazine, trimethoprim, piprad, nitrofurantoin, furazolidone, nalidixic acid, amiofloxacin, gatifloxacin, pazufloxacin, trofefloxacin, moxifloxacin acid, tetracyclines: tetracycline, methacycline, minocycline, aureomycin, doxycycline, oxytetracycline, doxycycline, metacycline, demeclocycline, guanadine, beta-lactamase inhibitor: clavulanic acid, sulbactam, triazolbactam, carbapenem antibiotics: imipenem, cilastatin, panipenem, betaminon, meropenem, cephalosporins, sulfonamides: sulfamone, sulfadimidine, sulfabenzzole, sulfamonomethoxine, iodoamine pair-methoxine, iodoamine doxine, sulfamidine, zinc sulfadiazine, sulfalin, succinsulfathiazole, bistrimethoprim, phthalothiazole, thiamycin, clavulanic acid, aztreonam, imipenem, faropenem, cilostatin, sulbactam, triazobactam, carrimonam, sisomicin, chloramphenicol palmitate, fosfomycin, SV, bromo Mo Pulin, octenidine, urotropine, meng Deli amine, bismuth subsalicylate, metronidazole disodium, shu Pai ketone, neo-sterilization, metronidazole, doxorubicin, epirubicin, pirarubicin, idarubicin, mupirocin, nitroimidazole, tinidazole, picolinic acid, nitrofurantoin, nitrofurans: furazolidone, trimethoprim, methylfurans: sulfasalazine, antifungal: sulconazole, lanoconazole, ji Nuokang azole, butoconazole, chloroconazole, fenticonazole nitrate, sertaconazole, oxiconazole, bifonazole, fluconazole, itraconazole, saperconazole, clotrimazole, econazole, tioconazole, miconazole, ketoconazole, naftifine, butenafine, ciclopirox, amorolfine, amphotericin B, clarithromycin, flucytosine, terbinafine, nystatin, griseofulvin, and clindamycin.
The 3- (2, 4-difluorophenyl) -2-methylpyrazolo [1,5-a ] pyrimidine-7-phenoxide derivatives and analogues of the structural formula I have pharmacological activity and application as antibacterial and antifungal medicaments, and the application of the derivatives and analogues in combination is characterized in that: the route of administration of the pharmaceutical compounds of the invention includes: administration is by oral, parenteral, subcutaneous, intravenous, intramuscular, intraperitoneal, transdermal, buccal, intrathecal, intracranial, intranasal, or topical routes.
Detailed Description
The present invention will be further illustrated by the following examples, which are merely illustrative of the present invention and are not intended to represent the scope of the invention as defined by the claims.
Chemical synthesis and preparation
(1) Preparation of important intermediates
3- (2, 4-difluorophenyl) -5-substituted-2-methylpyrazolo [1,5-a ] as shown in structural formula I]Pyrimidine-7-phenolate derivatives and analogues, their preparation method, characterized by: and (3) performing cyclization reaction under the action of a catalyst to prepare an intermediate 2, and reacting the carbonyl ethyl acetate compound containing various substituents with 4- (2, 4-difluorophenyl) -3-methyl-1H-pyrazol-5-amine under the action of the catalyst to form a C-N bond to complete the cyclization reaction. The catalyst is one of dehydrating agent, organic acid and/or inorganic acid catalyst, one of tetrahydrofuran, 1, 4-dioxane, acetonitrile, N-dimethylformamide, N-dimethylacetamide, N-hexane, toluene and the like is adopted as solvent, and the reaction temperature is controlled at-40 DEG C o C to 180 o Under the condition C, a key intermediate 2,3- (2, 4-difluorophenyl) -2-methylpyrazolo [1,5-a ] can be formed]Pyrimidine-7-ols of the formula:
(2) Preparation of phenoxide salts
3- (2, 4-difluorophenyl) -5-substituted-2-methylpyrazolo [1,5-a ]]The preparation method of pyrimidine-7-phenolate derivatives and analogues comprises the following steps: under the action of inorganic base, one of the following reagents (tetrahydrofuran, 1, 4-dioxane, acetonitrile, N-dimethylformamide, N-dimethylacetamide, etc.) is adopted as solvent, and the reaction temperature is controlled at-40 DEG C o C to 180 o Under the condition C, the phenolate compound 3 can be formed, and the reaction formula is as follows:
pharmaceutically acceptable salts of the compounds of the invention are also within the scope of the invention, the acids of which may be salified by reaction with bases, such as sodium carbonate, sodium hydride, potassium hydroxide, and the like. The structure containing nitrogen atoms has basicity and can be reacted with acid to form salts such as hydrochloric acid, fumaric acid, maleic acid, succinic acid, acetic acid, citric acid, tartaric acid, carbonic acid, phosphoric acid, oxalic acid and the like.
Prodrugs of the compounds of the present invention are also within the scope of the invention. The drug of the invention can be modified into a prodrug, which increases its water solubility and molecular volume and reduces its toxicity.
Examples of synthesis and preparation (examples and structures 1-33 are shown in Table 1)
Preparation of sodium 3- (2, 4-difluorophenyl) -2-methyl-5- (4- (trifluoromethyl) phenyl) pyrazolo [1,5-a ] pyrimidin-7-carboxylate of example 1
(1) In a 100 ml eggplant-shaped bottle, 0.209 g of 4- (2, 4-difluorophenyl) -3-methyl-1H-pyrazol-5-amine, 0.260 g of ethyl 3-oxo-3- (4- (trifluoromethyl) phenyl) propionate and 20. 20ml g of N, N-dimethylformamide were sequentially added at room temperature under nitrogen protection and stirring 120 o C for 5 hours, TLC detects completion of the reaction. Cooling, filtering, extracting with ethyl acetate three times (30 ml×2), backwashing with saturated saline twice (20 ml×2), drying with anhydrous sodium sulfate, recovering solvent under reduced pressure to obtain crude product, subjecting crude product to column chromatography to obtain crude solid, subjecting to silica gel column chromatography,by mobile phase CH 2 Cl 2 : CH 3 Gradient eluting with OH at different ratio concentrations to obtain eluate, and recrystallizing to obtain key intermediate 3- (2, 4-difluorophenyl) -2-methyl-5- (4- (trifluoromethyl) phenyl) pyrazolo [1, 5-a)]Pyrimidine-7-ols. IR (KBr, cm) -1 ) 3246, 2986, 1612, 1565, 1478, 1326, 1171, 1104, 1064, 1014, nuclear magnetic resonance spectrometry 1 H NMR (DMSO-d 6 ) δ11.53(b,1H), 7.75(d, J = 8.0, 1H), 7.72(d, J = 7.5, 2H), 7.68(d, J = 7.5, 2H), 7.07 (d, J = 8.0,1H), 6.82 (s,1H), 6.74 (s,1H), 2.06(s,3H)；
(2) In a 100 ml eggplant-shaped bottle, 3- (2, 4-difluorophenyl) -2-methyl-5- (4- (trifluoromethyl) phenyl) pyrazolo [1,5-a ] was sequentially added at room temperature ]Pyrimidine-7-sodium phenolate 0.427 g, sodium hydroxide 0.40 g, tetrahydrofuran and water (1:1) mixed solvent 20 ml, reflux reacting for 5 hours under nitrogen protection and stirring, recovering volume under reduced pressure, cooling, filtering to obtain crude solid, recrystallizing to obtain the target product, 3- (2, 4-difluorophenyl) -2-methyl-5- (4- (trifluoromethyl) phenyl) pyrazolo [1, 5-a)]Pyrimidine-7-ols. IR (KBr.cm-1) from 3659 to 2840, 1612, 1565, 1517, 1479, 1419, 1375, 1326, 1263, 1173, 1104, 1063, 1014, 964, 932, 851, 791, NMR spectroscopy 1 H NMR (DMSO-d 6 ) δ7.75(d, J = 8.0, 1H), 7.72(d, J = 7.5, 2H), 7.68(d, J = 7.5, 2H), 7.07 (d, J = 8.0,1H), 6.82 (s,1H), 6.74 (s,1H), 2.06(s, 3H)。
Example 2 preparation of sodium 5- (4-cyanophenyl) -3- (2, 4-difluorophenyl) -2-methylpyrazolo [1,5-a ] pyrimidin-7-ol
By adopting the similar preparation methods of (1) and (2) in example 1, respectively, 0.209 g of 4- (2, 4-difluorophenyl) -3-methyl-1H-pyrazol-5-amine and 0.217 g of 3- (4-cyanophenyl) -3-oxopropanoic acid ethyl ester were sequentially added to a eggplant-shaped bottle of 100 ml to react, to obtain an intermediate; and continuing to react with sodium hydroxide to obtain a target product. IR (KBr.cm-1) from 3661 to 2840, 2220, 1610, 1566, 1479, 1327, 1175, 1106, 1065, 964, NMR 1 H NMR (DMSO-d 6 ) δ7.97 (d, J = 7.5, 2H), 7.82 (d, J = 7.5, 2H), 7.75 (d, J = 8.0, 1H), 7.07 (d, J = 8.0, 1H), 6.82 (s, 1H), 6.74 (s, 1H), 2.06 (s, 3H)。
Example 3 preparation of sodium 3- (2, 4-difluorophenyl) -2-methyl-5- (4-nitrophenyl) pyrazolo [1,5-a ] pyrimidine-7-carboxylate
By adopting the similar preparation methods of (1) and (2) in example 1, respectively, 0.209 g of 4- (2, 4-difluorophenyl) -3-methyl-1H-pyrazol-5-amine and 0.237 g of 3- (4-nitrophenyl) -3-oxopropanoic acid ethyl ester were sequentially added to a eggplant-shaped bottle of 100 ml to react, to obtain an intermediate; and continuing to react with sodium hydroxide to obtain a target product. IR (KBr.cm-1) from 3664 to 2845, 1614, 1567, 1481, 1329, 1250, 1177, 1106, 1066, 1016, NMR 1 H NMR (DMSO-d 6 ) δ 8.32(d, J = 7.5, 2H), 8.05 (d, J = 7.5, 2H), 7.75 (d, J = 8.0, 1H), 7.07 (d, J = 8.0, 1H), 6.82 (s, 1H), 6.74 (s, 1H), 2.06 (s, 3H)。
Example 4 preparation of sodium 3- (2, 4-difluorophenyl) -5- (4-methoxyphenyl) -2-methylpyrazolo [1,5-a ] pyrimidin-7-ol
The intermediate was obtained by sequentially adding 0.209 g of 4- (2, 4-difluorophenyl) -3-methyl-1H-pyrazol-5-amine and 0.222 g g of ethyl 3- (4-methoxyphenyl) -3-oxopropanoate to a eggplant-shaped bottle of 100 ml by using the similar production methods of (1) and (2) in example 1, respectively; and continuing to react with sodium hydroxide to obtain a target product. IR (KBr.cm-1) from 3657 to 2838, 1610, 1563, 1477, 1324, 1250, 1171, 1102, 1061, 962, NMR 1 H NMR (DMSO-d 6 )
δ7.75 (d, J = 8.0, 1H), 7.55 (d, J = 7.5, 2H), 7.07 (m, 3H), 6.82 (s, 1H), 6.74 (s, 1H), 3.83 (s, 3H), 2.06 (s, 3H)。
Example 5 preparation of sodium 3- (2, 4-difluorophenyl) -5- (4- (dimethylamino) phenyl) -2-methylpyrazolo [1,5-a ] pyrimidin-7-carboxylate
Using the similar preparation methods of (1) and (2) in example 1, respectively, 0.209 g of 4- (2, 4-difluorophenyl) -3-methyl-1H-pyrazol-5-amine and 0.235 g of 3- (4- (dimethylamino) phenyl) -3-oxopropanoic acid ethyl ester were sequentially added to a 100 ml eggplant-shaped bottle to react, to obtain an intermediate; and continuing to react with sodium hydroxide to obtain a target product. IR (KBr.cm-1) determination by IR spectroscopy 3669-2837, 1 610, 1562, 1476, 1340, 1170, 1100, 1060, 1011; nuclear magnetic resonance spectrometry 1 H NMR (DMSO-d 6 )
δ7.75 (d, J = 8.0, 1H), 7.61 (d, J = 7.5, 2H), 7.07 (d, J = 8.0, 1H), 6.85 (d, J = 7.5, 2H), 6.82 (s, 1H), 6.74 (s, 1H), 3.06 (s, 6H), 2.06 (s, 3H)。
Example 6 preparation of sodium 3- (2, 4-difluorophenyl) -2-methyl-5- (4-phenolate sodium) -pyrazolo [1,5-a ] pyrimidine-7-carboxylate
By adopting the similar preparation methods of (1) and (2) in example 1, respectively, 0.209 g of 4- (2, 4-difluorophenyl) -3-methyl-1H-pyrazol-5-amine and 0.208 g g of 3- (4-hydroxyphenyl) -3-oxopropanoic acid ethyl ester were sequentially added to a eggplant-shaped bottle of 100 ml to react, to obtain an intermediate; and continuing to react with sodium hydroxide to obtain a target product. IR (KBr.cm-1) from 3658 to 2841, 1612, 1564, 1224, 1172, 1104, 1061, 1012, 960 from nuclear magnetic resonance spectrometry 1 H NMR (DMSO-d 6 ) δ7.75 (d, J = 8.0, 1H), 7.49 (d, J = 7.5, 2H), 7.07 (d, J = 8.0,1H), 6.86 (d, J = 7.5, 2H), 6.82 (s,1H), 6.74 (s, 1H), 2.06 (s, 3H)。
EXAMPLE 7 preparation of sodium 5- (4-carboxylate phenyl) -3- (2, 4-difluorophenyl) -2-methylpyrazolo [1,5-a ] pyrimidin-7-ol sodium
Using the similar preparation methods of (1) and (2) in example 1, respectively, 0.209 g of 4- (3-ethoxy-3-oxopropionyl) benzoic acid 0.236 g was added sequentially to a 100 ml eggplant-shaped bottle to react, thereby obtaining an intermediate; and continuing to react with sodium hydroxide to obtain a target product. IR (KBr.cm-1) from 3665 to 2810, 1617, 1600, 1480, 1326, 1173, 1104, 1063, 1014, NMR 1 H NMR (DMSO-d 6 ) δ8.10(d, J = 7.5, 2H), 8.0 (d, J = 7.5, 2H), 7.75 (d, J = 8.0, 1H), 7.07 (d, J = 8.0, 1H), 6.82 (s, 1H), 6.74 (s, 1H), 2.06 (s, 3H)。
Example 8 preparation of sodium 5- (4-sulfonate phenyl) -3- (2, 4-difluorophenyl) -2-methylpyrazolo [1,5-a ] pyrimidin-7-ol sodium
The preparation method similar to the preparation method (1) and (2) in the example 1 is adopted, and 4- (2, 4-difluoro) is added into a 100 ml eggplant-shaped bottle in sequencePhenyl) -3-methyl-1H-pyrazol-5-amine 0.209 g,4- (3-ethoxy-3-oxopropionyl) benzenesulfonic acid 0.256 g to afford intermediate; and continuing to react with sodium hydroxide to obtain a target product. IR (KBr.cm-1) from 3670 to 2810, 1612, 1565, 1479, 1326, 1200, 1104, 1063, 1014, NMR 1 H NMR (DMSO-d 6 )
δ8.01 (d, J = 7.5, 2H), 7.75 (d, J = 8.0, 1H)，7.64 (d, J = 7.5, 2H), 7.07 (d, J = 8.0, 1H), 6.82 (s, 1H), 6.74 (s, 1H), 2.06 (s, 3H)。
Example 9 preparation of sodium 3- (2, 4-difluorophenyl) -2-methyl-5- (4-sulfamoylphenyl) pyrazolo [1,5-a ] pyrimidin-7-carboxylate
Using the similar preparation methods of (1) and (2) in example 1, respectively, 0.209 g of ethyl 4- (2, 4-difluorophenyl) -3-methyl-1H-pyrazol-5-amine, 0.271 g g of 3-oxo-3- (4-sulfamoylphenyl) propionate was sequentially added to a eggplant-shaped bottle of 100 ml to react, to obtain an intermediate; and continuing to react with sodium hydroxide to obtain a target product. IR (KBr.cm-1) from 3665 to 2840, 1612, 1565, 1479, 1320, 1173, 1104, 1063, 1017, NMR 1 H NMR (DMSO-d 6 ) δ8.07 (d, J = 7.5, 2H), 7.92 (d, J = 7.5, 2H), 7.75 (d, J = 8.0, 1H), 7.07 (d, J = 8.0, 1H), 6.82 (s, 1H), 6.74 (s, 1H), 2.06(s, 3H), 2.0 (b, 2H)。
Example 10 preparation of sodium 3- (2, 4-difluorophenyl) -2-methyl-5- (4-phenoxyphenyl) pyrazolo [1,5-a ] pyrimidine-7-carboxylate
By adopting the similar preparation methods of (1) and (2) in example 1, respectively, and adding 0.209 g of 4- (2, 4-difluorophenyl) -3-methyl-1H-pyrazol-5-amine and 0.284 g g of 3-oxo-3- (4-phenoxyphenyl) propionic acid ethyl ester into a eggplant-shaped bottle of 100 ml in sequence, the reaction was carried out to obtain an intermediate; and continuing to react with sodium hydroxide to obtain a target product. IR (KBr.cm-1) from 3659 to 2840, 1612, 1565, 1479, 1326, 1173, 1104, 1063, 1014, NMR 1 H NMR (DMSO-d 6 ) δ7.60-7.07 (m, 11H), 6.82 (s, 1H), 6.74 (s, 1H), 2.06 (s, 3H)。
Example 11 preparation of sodium 5- (4-formylphenyl) -3- (2, 4-difluorophenyl) -2-methylpyrazolo [1,5-a ] pyrimidin-7-ol
By adopting the similar preparation methods of (1) and (2) in example 1, respectively, 0.209 g of 4- (2, 4-difluorophenyl) -3-methyl-1H-pyrazol-5-amine and 0.235 g of 3- (4-carbamoylphenyl) -3-oxopropanoic acid ethyl ester were sequentially added to a eggplant-shaped bottle of 100 ml to react, to obtain an intermediate; and continuing to react with sodium hydroxide to obtain a target product. IR (KBr.cm-1) from 3669 to 2840, 1750, 1565, 1479, 1326, 1173, 1104, 1063, 1014, NMR 1 H NMR (DMSO-d 6 )
δ8.09 (d, J = 7.5, 2H), 7.97 (d, J = 7.5, 2H), 7.75 (d, J = 8.0, 1H), 7.50 (b, 2H),7.07 (d, J = 8.0, 1H), 6.82 (s, 1H), 6.74 (s, 1H), 2.06 (s, 3H)。
Example 12 preparation of sodium 3, 5-bis (2, 4-difluorophenyl) -2-methylpyrazolo [1,5-a ] pyrimidin-7-carboxylate
Using the similar preparation methods of (1) and (2) in example 1, respectively, 0.209 g of 4- (2, 4-difluorophenyl) -3-methyl-1H-pyrazol-5-amine and 0.228 g of 3- (2, 4-difluorophenyl) -3-oxopropanoic acid ethyl ester were sequentially added to a eggplant-shaped bottle of 100 ml to react, to obtain an intermediate; and continuing to react with sodium hydroxide to obtain a target product. IR (KBr.cm-1) from 3669 to 2850, 1622, 1565, 1481, 1333, 1104, 1063, NMR 1 H NMR (DMSO-d 6 ) δ7.75 (d, J = 7.5, 2H), 7.07 (d, J = 7.5, 2H), 6.82 (s, 1H), 6.74 (s, 2H), 2.06 (s, 3H)。
EXAMPLE 13 preparation of sodium 5- (4-ethoxyphenyl) -3- (2, 4-difluorophenyl) -2-methylpyrazolo [1,5-a ] pyrimidin-7-ol
By adopting the similar preparation methods of (1) and (2) in example 1, respectively, 0.209 g of 4- (2, 4-difluorophenyl) -3-methyl-1H-pyrazol-5-amine and 0.250 g of 3- (4-acetoxyphenyl) -3-oxopropanoic acid ethyl ester were sequentially added to a eggplant-shaped bottle of 100 ml to react, to obtain an intermediate; IR (KBr.cm-1) for infrared spectrometry of target products, which is obtained by continuing to react with sodium hydroxide, 3652-2838, 1750, 1612, 1565, 1479, 1326, 1173, 1106, 1061, 1014, NMR spectrometry 1 H NMR (DMSO-d 6 )
δ7.83(d, J = 7.5, 2H), 7.75 (d, J = 8.0, 1H), 7.15 (d, J = 7.5, 2H), 7.07 (d, J = 8.0, 1H), 6.82 (s, 1H), 6.74 (s,1H), 2.28 (s, 3H), 2.06 (s, 3H)。
Example 14 preparation of sodium 3- (2, 4-difluorophenyl) -2-methyl-5- (6- (trifluoromethyl) pyridin-3-yl) pyrazolo [1,5-a ] pyrimidin-7-carboxylate
Using the similar preparation methods of (1) and (2) in example 1, respectively, 0.209 g of ethyl 4- (2, 4-difluorophenyl) -3-methyl-1H-pyrazol-5-amine and 0.261 g of 3-oxo-3- (6- (trifluoromethyl) pyridin-3-yl) propionic acid were sequentially added to a eggplant-shaped bottle of 100 ml to react, thereby obtaining an intermediate; and continuing to react with sodium hydroxide to obtain a target product. IR (KBr.cm-1) from 3659 to 2840, 1612, 1565, 1479, 1326, 1173, 1104, 1063, 1018, NMR 1 H NMR (DMSO-d 6 ) δ8.58 (s,1H), 7.88 (d, J = 7.5, 1H), 7.75 (d, J = 8.0, 1H), 7.35 (d, J = 7.5, 1H), 7.07 (d, J = 8.0, 1H), 6.82 (s, 1H), 6.74 (s, 1H), 2.06 (s, 3H)。
Example 15 preparation of sodium 5- (6-cyanopyridin-3-yl) -3- (2, 4-difluorophenyl) -2-methylpyrazolo [1,5-a ] pyrimidin-7-ol
Using the similar preparation methods of (1) and (2) in example 1, respectively, 0.209 g of 4- (2, 4-difluorophenyl) -3-methyl-1H-pyrazol-5-amine and 0.218 g of 3- (6-cyanopyridin-3-yl) -3-oxopropanoic acid ethyl ester were sequentially added to a 100 ml eggplant-shaped bottle to react, to obtain an intermediate; and continuing to react with sodium hydroxide to obtain a target product. IR (KBr.cm-1) from 3651 to 2831, 2240, 1612, 1565, 1479, 1326, 1173, 1104, 1063, 1017, NMR 1 H NMR (DMSO-d 6 ) δ9.20 (s, 1H), 8.35 (d, J = 7.5, 1H), 7.75 (d, J = 8.0, 1H), 7.73 (d, J = 7.5, 1H), 7.07 (d, J = 8.0, 1H), 6.82 (s, 1H), 6.74 (s, 1H), 2.06 (s, 3H)。
EXAMPLE 16 preparation of sodium 3- (2, 4-difluorophenyl) -2-methyl-5- (6-nitropyridin-3-yl) pyrazolo [1,5-a ] pyrimidin-7-carboxylate
The intermediate was obtained by sequentially adding 0.209 g of 4- (2, 4-difluorophenyl) -3-methyl-1H-pyrazol-5-amine and 0.238 g of ethyl 3- (6-nitropyridin-3-yl) -3-oxopropionate to a 100-ml eggplant-shaped bottle using the similar preparation methods of (1) and (2) in example 1, respectively; continuing to react with sodium hydroxideThe target product should be obtained. IR (KBr.cm-1) from 3651 to 2840, 1612, 1565, 1479, 1375, 1326, 1173, 1104, 1063, 1017, NMR 1 H NMR (DMSO-d 6 ) δ9.07 (s, 1H), 8.64 (d, J = 7.5, 1H), 8.15 (d, J = 7.5, 1H), 7.75 (d, J = 8.0, 1H), 7.07 (d, J = 8.0, 1H), 6.82 (s, 1H), 6.74 (s, 1H), 2.06 (s, 3H)。
Example 17 preparation of sodium 3- (2, 4-difluorophenyl) -5- (6-methoxypyridin-3-yl) -2-methylpyrazolo [1,5-a ] pyrimidin-7-carboxylate
By adopting the similar preparation methods of (1) and (2) in example 1, respectively, and adding 0.209 g of 4- (2, 4-difluorophenyl) -3-methyl-1H-pyrazol-5-amine and 0.223 g g of 3- (6-methoxypyridin-3-yl) -3-oxopropanoic acid ethyl ester into a eggplant-shaped bottle of 100 ml in sequence, the reaction was carried out to obtain an intermediate; and continuing to react with sodium hydroxide to obtain a target product. IR (KBr.cm-1) from 3649 to 2840, 1612, 1565, 1479, 1326, 1250, 1173, 1105, 1066, NMR 1 H NMR (DMSO-d 6 )
δ7.97 (d, J = 7.5, 1H), 7.91 (s, 1H), 7.75(d, J = 8.0, 1H)，7.07 (d, J = 8.0, 1H), 6.88 (s, 1H), 6.74 (s, 1H), 6.60 (d, J = 7.5, 1H), 3.80(s, 3H), 2.06(s, 3H)。
Example 18 preparation of sodium 3- (2, 4-difluorophenyl) -5- (6- (dimethylamino) pyridin-3-yl) -2-methylpyrazolo [1,5-a ] pyrimidin-7-carboxylate
Using the similar preparation methods of (1) and (2) in example 1, respectively, 0.209 g of 4- (2, 4-difluorophenyl) -3-methyl-1H-pyrazol-5-amine and 0.236 g of 3- (6- (dimethylamino) pyridin-3-yl) -3-oxopropanoic acid ethyl ester were sequentially added to a 100 ml eggplant-shaped bottle to react, to obtain an intermediate; IR (KBr.cm-1) for infrared spectrometry of target product by continuously reacting with sodium hydroxide, namely 3639-2850, 1612, 1565, 1479, 1326, 1200, 1104, 1063, 1021, and NMR spectrometry 1 H NMR (DMSO-d 6 )
δ8.33 (s, 1H), 7.75 (d, J = 8.0, 1H), 7.66 (d, J = 7.5, 1H), 7.07 (d, J = 8.0, 1H), 6.88 (s, 1H), 6.76 (d, J = 7.5, 1H), 6.74 (s, 1H), 3.15 (s, 6H), 2.06 (s, 3H)。
Example 19 preparation of sodium 3- (2, 4-difluorophenyl) -5- (6-phenolpyridin-3-yl) -2-methylpyrazolo [1,5-a ] pyrimidin-7-phenolate
By adopting the similar preparation methods of (1) and (2) in example 1, respectively, 0.209 g of 4- (2, 4-difluorophenyl) -3-methyl-1H-pyrazol-5-amine and 0.209 g g of 3- (6-hydroxypyridin-3-yl) -3-oxopropanoic acid ethyl ester were sequentially added to a eggplant-shaped bottle of 100 ml to react, to obtain an intermediate; IR (KBr.cm-1) for infrared spectrometry of target products, which is obtained by continuing to react with sodium hydroxide, 3659-2840, 1612, 1565, 1479, 1326, 1250, 1173, 1104, 1063, 1014, 964, NMR spectrometry 1 H NMR (DMSO-d 6 ) δ7.97 (d, J = 7.5, 1H), 7.91 (s,1H), 7.75 (d, J = 8.0, 1H), 7.07 (d, J = 8.0, 1H), 6.88 (s, 1H), 6.74 (s, 1H), 6.60 (d, J = 7.5, 1H), 2.06 (s, 3H)。
EXAMPLE 20 preparation of Potassium 5- (4-cyanophenyl) -3- (2, 4-difluorophenyl) -2-methylpyrazolo [1,5-a ] pyrimidin-7-ol
By adopting the similar preparation methods of (1) and (2) in example 1, respectively, 0.209 g of 4- (2, 4-difluorophenyl) -3-methyl-1H-pyrazol-5-amine and 0.217 g of 3- (4-cyanophenyl) -3-oxopropanoic acid ethyl ester were sequentially added to a eggplant-shaped bottle of 100 ml to react, to obtain an intermediate; and continuing to react with potassium hydroxide to obtain a target product. IR (KBr.cm-1) from 3661 to 2840, 2220, 1610, 1566, 1479, 1327, 1175, 1106, 1065, 964, NMR 1 H NMR (DMSO-d 6 ) δ7.97 (d, J = 7.5, 2H), 7.82 (d, J = 7.5, 2H), 7.75 (d, J = 8.0, 1H), 7.07 (d, J = 8.0, 1H), 6.82 (s, 1H), 6.74 (s, 1H), 2.06 (s, 3H)。
Example 21 preparation of sodium 5- (6-carboxylate pyridin-3-yl) -3- (2, 4-difluorophenyl) -2-methylpyrazolo [1,5-a ] pyrimidin-7-ol sodium
By adopting the similar preparation methods of (1) and (2) in example 1, respectively, and adding 0.209 g of 4- (2, 4-difluorophenyl) -3-methyl-1H-pyrazol-5-amine and 0.237 g g of 5- (3-ethoxy-3-oxopropionyl) pyridine carboxylic acid into a eggplant-shaped bottle of 100 ml in sequence, the reaction was carried out to obtain an intermediate; IR (KBr.cm-1) of target product obtained by continuing to react with sodium hydroxide, namely 3659-2840, 1660, 1565, 1479, 1326, 1173, 1104, 1063, 1020 of target product, and NMR of target product 1 H NMR (DMSO-d 6 ) δ9.23 (s, 1H), 8.40 (d, J = 7.5, 1H), 8.35 (d, J = 7.5, 1H), 7.75 (d, J = 8.0, 1H), 7.07 (d, J = 8.0, 1H), 6.88 (s, 1H), 6.74 (s, 1H), 2.06(s, 3H)。
Example 22 preparation of Potassium 3- (2, 4-difluorophenyl) -2-methyl-5- (4- (trifluoromethyl) phenyl) pyrazolo [1,5-a ] pyrimidin-7-ol
The preparation method similar to the preparation methods (1) and (2) in example 1 was adopted, respectively, and 3- (2, 4-difluorophenyl) -2-methyl-5- (4- (trifluoromethyl) phenyl) pyrazolo [1,5-a was sequentially added to a 100 ml eggplant-shaped bottle]Pyrimidine-7-phenol 0.427 g, potassium hydroxide 0.40 g, and reaction for 5 hours to obtain an intermediate; continuously reacting with potassium hydroxide to obtain a target product, namely 3- (2, 4-difluorophenyl) -2-methyl-5- (4- (trifluoromethyl) phenyl) pyrazolo [1, 5-a)]Pyrimidine-7-ols. IR (KBr.cm-1) from 3659 to 2840, 1612, 1565, 1479, 1326, 1173, 1104, 1063, 1014, 964, 932, 851, 791, NMR 1 H NMR (DMSO-d 6 ) δ7.75(d, J = 8.0, 1H), 7.72(d, J = 7.5, 2H), 7.68(d, J = 7.5, 2H), 7.07 (d, J = 8.0, 1H), 6.82 (s, 1H), 6.74 (s, 1H), 2.06(s, 3H)。
Example 23 preparation of Potassium 3- (2, 4-difluorophenyl) -2-methyl-5- (4-nitrophenyl) pyrazolo [1,5-a ] pyrimidin-7-ol
By adopting the similar preparation methods of (1) and (2) in example 1, respectively, 0.209 g of 4- (2, 4-difluorophenyl) -3-methyl-1H-pyrazol-5-amine and 0.237 g of 3- (4-nitrophenyl) -3-oxopropanoic acid ethyl ester were sequentially added to a eggplant-shaped bottle of 100 ml to react, to obtain an intermediate; and continuing to react with potassium hydroxide to obtain a target product. IR (KBr.cm-1) from 3664 to 2845, 1614, 1567, 1481, 1329, 1250, 1177, 1106, 1066, 1016, NMR 1 H NMR (DMSO-d 6 )
δ8.32(d, J = 7.5, 2H), 8.05 (d, J = 7.5, 2H), 7.75 (d, J = 8.0, 1H), 7.07 (d, J = 8.0, 1H), 6.82 (s, 1H), 6.74 (s, 1H), 2.06 (s, 3H)。
EXAMPLE 24 preparation of Potassium 3- (2, 4-difluorophenyl) -5- (4- (dimethylamino) phenyl) -2-methylpyrazolo [1,5-a ] pyrimidin-7-ol
The preparation method similar to the preparation method of (1) and (2) in the example 1 is adopted respectively, and the eggplant shape of the product is 100 mlBottle, add 0.209 g of 4- (2, 4-difluorophenyl) -3-methyl-1H-pyrazol-5-amine and 0.235. 0.235 g of 3- (4- (dimethylamino) phenyl) -3-oxopropanoic acid ethyl ester in sequence for reaction to obtain an intermediate; and continuing to react with potassium hydroxide to obtain a target product. IR (KBr.cm-1) from 3656 to 2837, 1610, 1562, 1476, 1340, 1170, 1100, 1060, 1011, NMR 1 H NMR (DMSO-d 6 )
δ7.75 (d, J = 8.0, 1H), 7.61(d, J = 7.5, 2H), 7.07 (d, J = 8.0, 1H), 6.85 (d, J = 7.5, 2H), 6.82 (s, 1H), 6.74 (s, 1H), 3.06 (s, 6H), 2.06 (s, 3H)。
EXAMPLE 25 preparation of Potassium 3- (2, 4-difluorophenyl) -2-methyl-5- (4-sulfamoylphenyl) pyrazolo [1,5-a ] pyrimidin-7-ol
Using the similar preparation methods of (1) and (2) in example 1, respectively, 0.209 g of ethyl 4- (2, 4-difluorophenyl) -3-methyl-1H-pyrazol-5-amine, 0.271 g g of 3-oxo-3- (4-sulfamoylphenyl) propionate was sequentially added to a eggplant-shaped bottle of 100 ml to react, to obtain an intermediate; and continuing to react with potassium hydroxide to obtain a target product. IR (KBr.cm-1) from 3665 to 2840, 1612, 1565, 1479, 1320, 1173, 1104, 1063, 1017, NMR 1 H NMR (DMSO-d 6 ) δ8.07 (d, J = 7.5, 2H), 7.92 (d, J = 7.5, 2H), 7.75 (d, J = 8.0, 1H), 7.07 (d, J = 8.0, 1H), 6.82 (s, 1H), 6.74 (s, 1H), 2.06(s, 3H), 2.0 (b, 2H)。
EXAMPLE 26 preparation of Potassium 3- (2, 4-difluorophenyl) -2-methyl-5- (6- (trifluoromethyl) pyridin-3-yl) pyrazolo [1,5-a ] pyrimidin-7-ol
Using the similar preparation methods of (1) and (2) in example 1, respectively, 0.209 g of ethyl 4- (2, 4-difluorophenyl) -3-methyl-1H-pyrazol-5-amine and 0.261 g of 3-oxo-3- (6- (trifluoromethyl) pyridin-3-yl) propionic acid were sequentially added to a eggplant-shaped bottle of 100 ml to react, thereby obtaining an intermediate; and continuing to react with potassium hydroxide to obtain a target product. IR (KBr.cm-1) from 3659 to 2840, 1612, 1565, 1479, 1326, 1173, 1104, 1063, 1018, NMR 1 H NMR (DMSO-d 6 ) δ8.58 (s,1H), 7.88(d, J = 7.5, 1H), 7.75(d, J = 8.0, 1H), 7.35(d, J = 7.5, 1H), 7.07 (d, J = 8.0, 1H), 6.82 (s, 1H), 6.74 (s,1H), 2.06 (s, 3H)。
EXAMPLE 27 preparation of 5- (6-cyanopyridin-3-yl) -3- (2, 4-difluorophenyl) -2-methylpyrazolo [1,5-a ] pyrimidin-7-ol potassium
Using the similar preparation methods of (1) and (2) in example 1, respectively, 0.209 g of 4- (2, 4-difluorophenyl) -3-methyl-1H-pyrazol-5-amine and 0.218 g of 3- (6-cyanopyridin-3-yl) -3-oxopropanoic acid ethyl ester were sequentially added to a 100 ml eggplant-shaped bottle to react, to obtain an intermediate; and continuing to react with potassium hydroxide to obtain a target product. IR (KBr.cm-1) from 3651 to 2831, 2240, 1612, 1565, 1479, 1326, 1173, 1104, 1063, 1017, NMR 1 H NMR (DMSO-d 6 ) δ9.20 (s, 1H), 8.35(d, J = 7.5, 1H), 7.75 (d, J = 8.0, 1H), 7.73(d, J = 7.5, 1H), 7.07 (d, J = 8.0, 1H), 6.82 (s, 1H), 6.74 (s, 1H), 2.06 (s, 3H)。
EXAMPLE 28 preparation of sodium 3- (2, 4-difluorophenyl) -2-methyl-5- (6-sulfinate pyridin-3-yl) pyrazolo [1,5-a ] pyrimidin-7-ol
By adopting the similar preparation methods of (1) and (2) in example 1, respectively, 0.209 g of 4- (2, 4-difluorophenyl) -3-methyl-1H-pyrazol-5-amine and 0.257 g of 5- (3-ethoxy-3-oxopropionyl) pyridine-2-sulfinic acid were sequentially added to a eggplant-shaped bottle of 100 ml to react, to obtain an intermediate; IR (KBr.cm-1) for infrared spectrometry of target products, which is obtained by continuing to react with sodium hydroxide, namely 3666-2840, 1612, 1565, 1479, 1326, 1253, 1173, 1104, 1063 and 1016, nuclear magnetic resonance spectrometry 1 H NMR (DMSO-d 6 ) δ8.19 (d, J = 7.5, 1H), 7.83 (d, J = 7.5, 1H), 7.75 (d, J = 8.0, 1H), 7.07 (d, J = 8.0, 1H), 6.88 (s, 1H), 6.74 (s,1H) , 2.06 (s, 3H), 2.0 (b, 1H)。
Example 29 preparation of sodium 3- (2, 4-difluorophenyl) -2-methyl-5- (6-sulfamoylpyridin-3-yl) pyrazolo [1,5-a ] pyrimidin-7-carboxylate
Using the similar preparation methods of (1) and (2) in example 1, respectively, 0.209 g of ethyl 4- (2, 4-difluorophenyl) -3-methyl-1H-pyrazol-5-amine, 0.272 g g of 3-oxo-3- (6-sulfamoylpyridin-3-yl) propionate was sequentially added to a eggplant-shaped bottle of 100 ml to react, to obtain an intermediate; and continuing to react with sodium hydroxide to obtain a target product. IR Spectrometry (KBr)Cm-1) 3651-2840, 1612, 1565, 1479, 1330, 1173, 1063, 1021, nuclear magnetic resonance spectrometry 1 H NMR (DMSO-d 6 ) δ9.24 (s,1H), 8.42 (d, J = 7.5, 1H), 7.75 (d, J = 8.0, 1H), 7.57 (d, J = 7.5, 1H), 7.07 (d, J = 8.0, 1H), 6.88 (s, 1H), 6.74 (s,1H) , 2.06 (s, 3H), 2.0 (b, 2H)。
EXAMPLE 30 preparation of sodium 3- (2, 4-difluorophenyl) -2-methyl-5- (6-phenoxypyridin-3-yl) pyrazolo [1,5-a ] pyrimidin-7-carboxylate
By adopting the similar preparation methods of (1) and (2) in example 1, respectively, and adding 0.209 g of 4- (2, 4-difluorophenyl) -3-methyl-1H-pyrazol-5-amine and 0.285 g g of 3-oxo-3- (6-phenoxypyridin-3-yl) propionic acid ethyl ester into a eggplant-shaped bottle of 100 ml in sequence, the reaction was carried out to obtain an intermediate; and continuing to react with sodium hydroxide to obtain a target product. IR (KBr.cm-1) from 3639 to 2860, 1612, 1565, 1479, 1326, 1255, 1173, 1104, 1063, 1009, NMR 1 H NMR (DMSO-d 6 ) δ7.96-6.9 (m, 10H), 6.88 (s, 1H), 6.74 (s, 1H), 2.06 (s, 3H)。
Example 31 preparation of sodium 3- (2, 4-difluorophenyl) -5- (4- ((2- (dimethylamino) ethyl) carbamoyl) phenyl) -2-methylpyrazolo [1,5-a ] pyrimidin-7-carboxylate
Using the similar preparation methods of (1) and (2) in example 1, respectively, 0.209 g of 4- (2, 4-difluorophenyl) -3-methyl-1H-pyrazol-5-amine and 0.306 g of 3- (4- ((2- (dimethylamino) ethyl) carbamoyl) phenyl) -3-oxopropanoic acid ethyl ester were sequentially added to a 100 ml eggplant-shaped bottle to react, to obtain an intermediate; and continuing to react with sodium hydroxide to obtain a target product. IR (KBr.cm-1) from 3656 to 2837, 1680, 1640, 1562, 1476, 1410, 1340, 1170, 1100, 1060, 1011, NMR 1 H NMR (DMSO-d 6 ) δ8.09(d, J = 7.5, 3H) 7.97(d, J = 7.5, 2H), 7.75(d, J = 8.0, 1H), 7.07 (d, J = 8.0,1H), 6.82 (s, 1H), 6.74 (s, 1H), 3.60 (t, J = 7.5, 2H), 2.57 (t, J = 7.5, 2H), 2.26 (s, 6H), 2.06 (s,3H)。
EXAMPLE 32 preparation of sodium 5- (6-formylpyridin-3-yl) -3- (2, 4-difluorophenyl) -2-methylpyrazolo [1,5-a ] pyrimidin-7-ol
Respectively using those in example 1The similar preparation method of (1) and (2), adding 0.209 g of 4- (2, 4-difluorophenyl) -3-methyl-1H-pyrazol-5-amine and 0.236 g g of 3-oxo-3- (6-phenoxypyridin-3-yl) ethyl propionate into a eggplant-shaped bottle of 100 ml in sequence for reaction to obtain an intermediate; and continuing to react with sodium hydroxide to obtain a target product. IR (KBr.cm-1) from 3659 to 2840, 1720, 1565, 1479, 1326, 1173, 1104, 1063, 1021, NMR 1 H NMR (DMSO-d 6 )
δ9.11 (s, 1H), 8.44 (d, J = 7.5, 1H), 8.41 (d, J = 7.5, 1H), 8.37 (b, 2H), 7.75 (d, J = 8.0, 1H), 7.07 (d, J = 8.0, 1H), 6.88 (s, 1H), 6.74 (s, 1H), 2.06 (s, 3H)。
Example 33 preparation of sodium 5- (6-Acetoxypyridin-3-yl) -3- (2, 4-difluorophenyl) -2-methylpyrazolo [1,5-a ] pyrimidin-7-ol
Using the similar preparation methods of (1) and (2) in example 1, respectively, 0.209 g of ethyl 4- (2, 4-difluorophenyl) -3-methyl-1H-pyrazol-5-amine, 0.251 g g of 3- (6-acetoxypyridin-3-yl) -3-oxopropanoate was sequentially added to a 100 ml eggplant-shaped bottle to react, to obtain an intermediate; and continuing to react with sodium hydroxide to obtain a target product. IR (KBr.cm-1) from 3622 to 2830, 1730, 1565, 1479, 1326, 1173, 1104, 1065, NMR 1 H NMR (DMSO-d 6 ) δ7.97 (d, J = 7.5, 1H), 7.91 (s, 1H), 7.75 (d, J = 8.0, 1H), 7.07 (d, J = 8.0, 1H), 6.88 (s, 1H), 6.74 (s, 1H), 6.60 (d, J = 7.5, 1H), 2.28 (s, 3H), 2.06 (s, 3H)。
Table 1 example compounds
Examples Structural formula Molecular formula Molecular weight
1 C 20 H 11 F 5 N 3 NaO 427.07
2 C 20 H 11 F 2 N 4 NaO 384.08
3 C 19 H 11 F 2 N 4 NaO 3 404.07
4 C 20 H 14 F 2 N 3 NaO 2 389.10
5 C 21 H 17 F 2 N 4 NaO 402.13
6 C 19 H 11 F 2 N 3 Na 2 O 2 397.06
7 C 20 H 11 F 2 N 3 Na 2 O 3 425.06
8 C 19 H 11 F 2 N 3 Na 2 O 3 S 445.03
9 C 19 H 13 F 2 N 4 NaO 3 S 438.06
10 C 25 H 16 F 2 N 3 NaO 2 451.11
11 C 20 H 13 F 2 N 4 NaO 2 402.09
12 C 19 H 10 F 4 N 3 NaO 395.07
13 C 21 H 14 F 2 N 3 NaO 3 417.09
14 C 19 H 10 F 5 N 4 NaO 428.07
15 C 19 H 10 F 2 N 5 NaO 385.08
16 C 18 H 10 F 2 N 5 NaO 3 405.06
17 C 19 H 13 F 2 N 4 NaO 2 390.09
18 C 20 H 16 F 2 N 5 NaO 403.12
19 C 18 H 10 F 2 N 4 Na 2 O 2 398.06
20 C 20 H 11 F 2 KN 4 O 400.05
21 C 19 H 10 F 2 N 4 Na 2 O 3 426.05
22 C 20 H 11 F 5 KN 3 O 443.05
23 C 19 H 11 F 2 KN 4 O 3 420.04
24 C 21 H 17 F 2 KN 4 O 418.10
25 C 19 H 13 F 2 KN 4 O 3 S 454.03
26 C 19 H 10 F 5 KN 4 O 444.04
27 C 19 H 10 F 2 KN 5 O 401.05
28 C 18 H 10 F 2 N 4 Na 2 O 3 S 446.02
29 C 18 H 12 F 2 N 5 NaO 3 S 439.05
30 C 24 H 15 F 2 N 4 NaO 2 452.11
31 C 24 H 22 F 2 N 5 NaO 2 473.16
32 C 19 H 12 F 2 N 5 NaO 2 403.09
33 C 20 H 13 F 2 N 4 NaO 3 418.09
Example 34 in vitro antibacterial Experimental example
Examples of injection preparation
Weighing 0.50 g compound 1, adding ethanol 60 ml, stirring to dissolve, adding 60 ml of 1, 2-propylene glycol and 10 ml Tween 80 after dissolving, stirring and mixing uniformly, adding water for injection to total volume of 500 ml, filtering with 0.22 μm filter membrane, subpackaging, sterilizing at 100deg.C under hot pressure for 30 min, detecting leak, performing total detection, and packaging to obtain 5 mg/5 ml (ammonia bottle) for 100 pieces.
Materials and methods
1. Standard strain: bacillus cereus 2, bacillus subtilis 168, enterococcus faecalis 29212, enterococcus faecalis F2518 (vre), staphylococcus aureus 29231, staphylococcus aureus 43300 (MRSA), staphylococcus aureus 703 (MRSA), streptococcus pneumoniae 6303 (PRSP), streptococcus pyogenes M2, streptococcus agalactiae group B, streptococcus pneumoniae 10351, bacillus anthracis 1, bacillus diphtheriae, clostridium perfringens, candida stomatitis, dermatophytes;
2. Test sample: compound 1, compound 2, compound 3, compound 4, compound 5, compound 6, compound 7, compound 8, compound 9, compound 10, compound 11, compound 12, compound 13, compound 14, compound 15, compound 16, compound 17, compound 18, compound 19, compound 20, compound 21, compound 22, compound 23, compound 24, compound 25, compound 26, compound 27, compound 28, compound 29, compound 30, compound 31, compound 32, compound 33
3. Method of
(1) And (3) sterilization: sterilizing the required experimental equipment and culture solution at 121deg.C for 30min under high pressure; ultraviolet irradiation for 30min in a sterile room;
(2) Bacteria increasing bacteria
Preparation of broth medium: accurately weighing 6g of pancreatic protein soybean broth culture medium in a 500ml beaker, adding 200ml distilled water, heating to dissolve thoroughly, transferring into a triangular flask, adding a cotton plug, wrapping, and sterilizing under high pressure;
preparation of a slant culture medium: weighing 3.8g of pancreatic protein soybean agar in a 500ml beaker, adding 100ml of distilled water, heating for complete dissolution, transferring into a triangular flask, adding a cotton plug, and packaging for high-pressure sterilization. After cooling, sub-packaging in 7 test tubes, wherein each test tube is about 10-15 ml, and the test tube is inclined at a proper angle and cooled for standby;
Bacterial amplification: the ATCC 4300 sealed vial was opened, a small amount of the bacterial powder was collected with a small sterilized ophthalmic forceps, transferred into a 5ml centrifuge tube, and added with 0.6ml of the pancreatic protein soybean broth medium, and mixed well. Average to 7 slant culture medium, namely 80 μl/tube, and uniformly coating. Placing the mixture into an incubator, and culturing for 24 hours at 37 ℃;
bacterial suspension preparation and bacterial count: the bacteria in the large tube were washed off with culture medium and transferred to a sterile centrifuge tube. Uniformly mixing to obtain bacterial suspension. A drop was removed from the pipette and bacterial density was measured. A piece of clean blood cell counting plate is taken, a cover glass is covered on a counting area, the taken bacterial suspension is diluted by a certain multiple by normal saline and is blown uniformly, a small amount of liquid is sucked by a liquid-transferring gun, and a drop (not too much) is dripped into grooves on two sides of a middle platform of the counting plate along the lower edge of the cover glass, so that the bacterial suspension fills the counting area by utilizing the surface tension of liquid, and no bubbles are generated. The plates were placed under a microscope for bacterial count. The total number of bacteria in 16 cells was counted, and the bacterial liquid concentration was calculated using the following formula:
bacterial Density (number/ml) =16 cell bacterial count×10 4 X dilution factor
(3) Inoculation of 96-well plates
Sample preparation: firstly, fully dissolving a drug to be tested with a small amount of DMSO, preparing the drug to be tested into a required initial concentration by using a culture medium, and sequentially diluting the drug to be tested into each gradient;
preparing bacterial liquid: based on the measurement result of the bacterial concentration, the culture solution (TSB) for bacterial suspension was diluted to 1.07X 10 7 Bacteria liquid with cfμ/ml concentration;
dosing scheme: the experiment is divided into a positive control group, a normal saline group, a blank control group and each test drug group, wherein each test drug group, the normal saline group and the blank group are respectively provided with 6 gradient holes, and the positive control group is provided with 7 gradient holes. 50 μl of the bacterial suspension, 30 μl of the culture solution and 20 μl of each sample solution were sequentially added to each well;
culturing and observing: the 96-well plate after sample addition is placed in an incubator for culture. The incubation temperature was 37℃and the incubation time was 24 hours. After the culture was completed, the growth of each colony was observed in the ultra clean bench. The bacterial liquid is clear, free from turbidity and free from bacterial colony at the bottom of the hole. The concentration was defined as the Minimum Inhibitory Concentration (MIC) of the drug.
Results and conclusions
The stock solution is diluted by a micro dilution method on a microporous dilution plate of a 96-well plate (1 st to 10 th holes), 50 mu L of bacterial solution is added into the 1 st to 11 th holes, the concentration of the liquid solution in the 1 st to 10 th holes is reduced by times, and 100 mu L of culture medium is added into the 12 th holes to serve as a blank control. After shaking and mixing the microporous dilution plates, placing the plates in a covered porcelain plate filled with wet gauze, culturing the plates for 24 hours at 37 ℃, and observing the results under a light source with a black background. The bacteria grow in a ball shape, diffuse turbidity or the bottom is in a button-like sediment, and the lowest drug concentration contained in the sterile growth hole is the lowest antibacterial concentration. The results are shown in Table 2 and Table 2 (subsequent Table)
And (3) injection:
(1) Bacteria and fungi (with the same is drug-resistant bacteria)
1. Bacillus cereus 2;2. bacillus subtilis 168;3. enterococcus faecalis 29212; enterococcus faecalis F2518 (VRE); 5. staphylococcus aureus 29231; staphylococcus aureus 43300 (MRSA); staphylococcus aureus 703 (MRSA); streptococcus pneumoniae 6303 (PRSP); 9, streptococcus pyogenes M2;10. group B streptococcus agalactiae; 11. streptococcus pneumoniae 10351;12. bacillus anthracis 1;13. diphtheria bacillus; 14, perfringens; 15. candida stomatitis; 16. dermatophytes;
(2) MIC value (. Mu.M) measurement values
-representing MIC values (μm) >150 μg/mL, + representing MIC values (μm) < 150 μg/mL, ++ representing MIC values (μm) < 100 μg/mL, ++ representing MIC values (μm) < 50 μg/mL;
conclusion:
(1) Table 2 shows that the compounds of the examples of the present invention have excellent antibacterial activity. Of particular note are MIC values (μm) for example compounds 2 and 3 for staphylococcus aureus 43300 (MRSA) and for staphylococcus aureus 703 (MRSA) are < 50 μg/mL, respectively;
(2) The compounds 1,2,3,4,6,7,9,10,11,12,13,14,15,16,17,18,19,20,21,22,23,24,25,26,27,29,30,31,32 and 33 respectively have inhibiting effects on a plurality of different resistant bacteria, such as enterococcus faecalis F2518 (VRE), staphylococcus aureus 43300 (MRSA), staphylococcus aureus 703 (MRSA) and streptococcus pneumoniae 6303 (PRSP), and MIC values (mu M) are respectively in the interval of < 100 mu g// mL;
(3) Compounds 1,2,3,6,9,10,14,15,16,20,22,24,26,27 and 31 were effective against bacillus cereus 2, bacillus subtilis 168, enterococcus faecalis 29212, enterococcus faecalis F2518 (VRE), staphylococcus aureus 29231, staphylococcus aureus 43300 (MRSA), staphylococcus aureus 703 (MRSA), streptococcus pneumoniae 6303 (PRSP) and streptococcus pyogenes M2, with MIC values (μm) in the < 100 μg// mL interval, respectively;
(4) Compounds 1,5,6,9,13,14, 16,17,21,22,23,24,25,26,27,28,29,30,31,32 and 33 were effective against oral candida, with MIC values (μm) in the < 100 μg// mL interval, respectively.
EXAMPLE 35 in vivo antibacterial Experimental example
1. Material
Test sample: compound 3, compound 5, compound 9, compound 10, compound 30, compound 1, compound 2;
test animals: healthy mice of Kunming species, weight 19-21 g, male and female half groups, other groups are used by single sex, provided by animal center of medical institute of Beijing medical science academy of sciences;
strains: staphylococcus aureus 43300 (MRSA);
2. method of
Mice were randomly divided into a blank control group, a positive control group, and a test drug group, 10 mice per group, and male and female halves. Inoculating abdominal cavity bacteria (MRSA-2152) at a concentration of 5.0X10 according to weight of 0.2ml/10g 6 cfu/ml, tail vein injection administration was performed immediately after inoculation, and a second administration was performed after 6 hours. After observation for 30 days, the survival time of each group of animals is recorded, and the life extension rates of the positive control group and the sample group are calculated:
percent life extension = (subject day of survival-day of survival in blank)/day of survival in blank x 100%
3. Results and conclusions
Note that: compound 1 is compound 30+ciprofloxacin with the ratio of 1:1; compound 2 is compound 5+ ciprofloxacin with the ratio of 1:1
In vivo experiments show that the compound 3, the compound 5, the compound 9, the compound 10, the compound 30, the compound 1 and the compound 2 have certain inhibition effect on drug-resistant staphylococcus aureus 43300 (MRSA), the life extension rate exceeds 50% when in administration, and the life extension rate of the compound is obviously improved compared with that of a single prescription, so that the life extension rate of the compound can be obviously enhanced when the compound is used. The above samples can be used as a further in-depth study against MRSA.

Claims (6)

1. 3- (2, 4-difluorophenyl) -5-substituted-2-methylpyrazolo [1,5-a ] pyrimidine-7-phenoxide derivative, characterized in that the derivative has at least one of the following structures:
2. use of a 3- (2, 4-difluorophenyl) -5-substituted-2-methylpyrazolo [1,5-a ] pyrimidine-7-phenoxide derivative according to claim 1 for the preparation of a medicament having antibacterial activity, wherein the bacterium is a gram-positive bacterium: staphylococci, enterococcus faecalis, streptococcus bovis, streptococcus pneumoniae, streptococcus agalactiae, diphtheria bacillus, bacillus anthracis, bacillus cereus, and Bacillus perfringens.
3. The use according to claim 2, wherein the bacteria are gram-positive bacteria, which are: streptococcus pyogenes.
Use of sodium 3- (2, 4-difluorophenyl) -2-methyl-5- (4-nitrophenyl) pyrazolo [1,5-a ] pyrimidine-7-carboxylate in the preparation of a medicament having antibacterial activity, wherein the bacteria are staphylococcus aureus 43300 and/or staphylococcus aureus 703.
5. The use according to any one of claims 2 to 4, wherein the medicament is administered by the route of: at least one of oral, intravenous, intramuscular, intraperitoneal, transdermal.
6. The use according to any one of claims 2 to 4, wherein the medicament is administered by the route of: at least one of buccal, intrathecal, intracranial, intranasal.
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