Pyrazole cyanoethylene pyridine ketonic acid compound as well as preparation method and medical application thereof

Abstract

The invention relates to a pyrazol cyanogen vinyl pyridine ketonic acid compound and a preparation method and medical application thereof, and belongs to the technical field of chemical synthesis, the pyrazol cyanogen vinyl pyridine ketonic acid compound is shown in general formulas I-III, and the compound has good inhibitory activity on one or more of gram-positive bacteria and gram-negative bacteria, can be used for preparing antibacterial drugs, and can be used for preparing antibacterial drugs. More efficient and safe candidate drugs are provided for clinical anti-infection treatment, and clinical treatment problems such as increasingly serious microbial drug resistance, stubborn pathogenic microorganisms and newly appearing harmful microorganisms can be solved.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of chemical synthesis, and particularly relates to a pyrazole cyanovinylpyridone acid compound, a preparation method thereof and medical application. BACKGROUND

[0002] The widespread use and even abuse of antibiotics has led to a rapid increase in antibiotic resistance, and there is an urgent need to develop new anti-infective drugs, especially therapeutic drugs for drug-resistant microbial strains.

[0003] Five-membered aromatic heterocyclic pyrazole has a planar conjugated structure, and six delocalized π electrons endow it with aromatic characteristics. At the same time, due to the different electronic properties of the pyrrole-like nitrogen atom and the pyridine-like nitrogen atom of pyrazole, different anions and cations can be obtained under different acid-base conditions, so that pyrazole can interact with biological targets through different non-covalent bonds, such as π-π stacking, π-cation (anion), hydrogen bonding, etc., to produce strong and effective biological effects. Since last century, many pyrazole derivatives with various biological activities have been developed, which have shown great potential in the agricultural chemical industry, making pyrazole an important structural unit for drug discovery and development. However, in recent years, the research on the pharmacological activity of pyrazoles has mainly focused on anti-inflammatory, anticancer, antiviral, etc., and relatively less on antibacterial. Literature reports that pyrazole derivatives can inhibit microbial growth and overcome drug resistance through various modes of action, such as inhibiting the synthesis of DNA, cell wall and protein in the strain, damaging the cell membrane, etc., which reveals the great potential of pyrazole compounds in combating the crisis of microbial drug resistance.

[0004] Quinolones are one of the most commonly used antibacterial drugs in clinic, which have good pharmacokinetic properties, high oral bioavailability and excellent efficacy. These drugs interact with DNA gyrase or DNA topoisomerase IV to induce irreparable DNA breakage, and exert strong antibacterial efficacy. However, with the long-term and extensive use of this class of drugs, target gene mutations, membrane permeability changes caused by pathogen defense system regulation and outer membrane protein changes have led to the rapid evolution and widespread distribution of drug-resistant strains, and there is an urgent need to develop new antibacterial drugs with novel structures. Previous studies have shown that modification of various important drug fragments at the C-7 position of the quinolone skeleton is a promising strategy that can bypass antibiotic resistance mechanisms and maintain inhibitory potency against multi-drug resistant bacteria. In particular, many azole-modified quinolones have excellent antibacterial effects, low drug resistance and good biocompatibility. In this study, different linking fragments were used at the C-7 position of the quinolone to covalently link the aromatic pyrazole ring to the core framework of the quinolone drug, obtaining pyrazole cyanovinylpyridone acid molecules with a new framework structure. The new framework structure molecules may interact with the target through various non-covalent forces to improve the affinity of the compound with DNA, enzymes or other receptors in the body, and enhance the antibacterial activity. At the same time, the construction of new structure molecules may lead to the discovery of new antibacterial mechanisms, which is expected to overcome serious bacterial resistance. SUMMARY

[0005] Therefore, one of the purposes of the present application is to provide pyrazole cyanovinylpyridone acid compounds and their pharmaceutically acceptable salts; the second purpose is to provide a preparation method of pyrazole cyanovinylpyridone acid compounds and their pharmaceutically acceptable salts; the third purpose is to provide the application of pyrazole cyanovinylpyridone acid compounds and their pharmaceutically acceptable salts in the preparation of antibacterial drugs, so as to provide more efficient and safe candidate drugs for clinical antibacterial infection treatment, and help to solve the increasingly serious drug resistance, stubborn pathogenic microorganisms and newly emerging harmful microorganisms and other clinical treatment problems.

[0006] In order to achieve the above-mentioned purposes, the present application provides the following technical solutions:

[0007] 1. Pyrazole cyanovinylpyridone acid compounds and their pharmaceutically acceptable salts, the structure is shown in general formula I and II:

[0008]

[0009] In the formula,

[0010] n is an integer from 0 to 17;

[0011] R 1 is alkyl, alkenyl, alkynyl, cycloalkyl and heterocycloalkyl, hydroxyl, cyano, ester, carboxyl, phenyl, substituted phenyl or heteroaryl;

[0012] R 2 , R3 It can be hydrogen, halogen, or alkyl.

[0013] Preferred:

[0014] n can be any one of 0, 1 or 3;

[0015] R 1 It is any one of alkyl, alkenyl, cycloalkyl, phenyl, substituted phenyl, or heteroaryl;

[0016] R 2 R 3 It is any one of hydrogen, halogen, or alkyl.

[0017] Preferably, it is any one of the following compounds:

[0018]

[0019] Preferably, the pharmaceutically acceptable salt is a hydrochloride, bromate, iodate, sulfate, nitrate, trifluoroacetate, or acetate.

[0020] 2. The method for preparing the pyrazolium cyanide pyridinone acid compound and its pharmaceutically acceptable salt, wherein the method is as follows:

[0021] a) Compounds of general formulas IV and V are added to ethanol, followed by the addition of various pyrazole aldehydes, and then refluxed under the action of a base to obtain pyrazole cyanide vinylpyridinone acid compounds of general formulas I and II.

[0022]

[0023] in:

[0024] n is 0, 1, or 3.

[0025] R 1 It can be alkyl, alkenyl, cycloalkyl, phenyl, substituted phenyl, or heteroaryl;

[0026] b. Add the compound shown in general formula VI to ethanol, add various pyrazole aldehyde compounds, and then reflux the reaction under the action of acid to obtain the pyrazole cyanide pyridinone acid compound shown in general formula III.

[0027]

[0028] c. Preparation of pharmaceutically acceptable salts of pyrazolium cyanide pyridinone acid compounds represented by general formulas I to III: Dissolve the pyrazolium cyanide pyridinone acid compounds represented by general formulas I to III in an organic solvent, add a pharmaceutically acceptable acid and react until no precipitate is formed, thus obtaining pharmaceutically acceptable salts of pyrazolium cyanide pyridinone acid compounds represented by formulas I to III.

[0029] Preferably,

[0030] In step a, the base is piperidine, the molar ratio of intermediate IV or V, pyrazole aldehyde compound and base is 1:1-2:0.2-1, and the reaction is carried out in ethanol at 60-90°C for 8-24 hours.

[0031] In step b, the acid is glacial acetic acid, the molar ratio of intermediate VI, pyrazole aldehyde compound and acid is 1:1-2:0.2-0.5, and the reaction is carried out in ethanol at 60-90°C for 1-4 hours.

[0032] In step c, the organic solvent is at least one of chloroform, acetone, acetonitrile, diethyl ether, tetrahydrofuran, dioxane or N,N-dimethylformamide; and the pharmaceutically acceptable acid is hydrochloric acid or sulfuric acid.

[0033] 3. Use of the pyrazole cyanovinylpyridone acid compound and its pharmaceutically acceptable salt in the preparation of an antibacterial drug.

[0034] Preferably, the bacteria are any one or more of methicillin-resistant Staphylococcus aureus ATCC 43300, Staphylococcus aureus ATCC 12600, Staphylococcus aureus ATCC 25923, Staphylococcus aureus ATCC 29213, Enterococcus faecalis p1-2007226001, Streptococcus pyogenes ATCC 19615, Klebsiella pneumoniae CMCC(B)46117, Escherichia coli ATCC 8739, Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 15442, Pseudomonas aeruginosa ATCC 27853 or Acinetobacter baumannii ATCC 19606.

[0035] 4. A preparation containing the pyrazole cyanovinylpyridone acid compound and its pharmaceutically acceptable salt.

[0036] Preferably, the preparation is one of a tablet, a capsule, a granule, an injection, a powder injection, an eye drop, a liniment, a suppository, an ointment or an aerosol.

[0037] The pyrazole cyanoethylene pyridone acid compound, the preparation method and the application thereof are provided in the present application, the five-membered aromatic heterocyclic ring pyrazole is hybridized with the important functional fragment such as cyanoethylene and hydrazone at the C-7 position of quinolone by using the drug design splicing principle, a series of pyrazole cyanoethylene pyridone acid compounds are designed and synthesized, the compounds are found to have certain inhibitory activity on gram-positive bacteria (methicillin-resistant Staphylococcus aureus ATCC 43300, Staphylococcus aureus ATCC 12600, Staphylococcus aureus ATCC 25923, Staphylococcus aureus ATCC 29213, Enterococcus faecalis p1-2007226001, Streptococcus pyogenes ATCC 19615) and gram-negative bacteria (Klebsiella pneumoniae CMCC (B) 46117, Escherichia coli ATCC 8739, Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 15442, Pseudomonas aeruginosa ATCC 27853, Acinetobacter baumannii ATCC 19606), and can be used for preparing antibacterial drugs, providing more efficient and safe candidate drugs for clinical antibacterial treatment, and helping to solve the increasingly serious drug resistance, stubborn pathogenic microorganisms and new emerging harmful microorganisms and other clinical treatment problems.

[0038] Other advantages, objects, and features of the present application will be better understood from the following specification taken in conjunction with the accompanying drawings. It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not intended to be restrictive of the present application, as claimed. DETAILED DESCRIPTION

[0039] Other advantages and benefits of the present application will become apparent to those skilled in the art, upon perusal of the following specification. The present application can be put into practice by means and methods other than those specifically described herein, and the details of construction and the arrangements of parts herein described are not limited to the specifically described embodiments.

[0040] Preparation of Example 1, intermediates IV and V

[0041]

[0042] Intermediate IV and V were prepared according to the method described in the reference “[1] Kamal, A.; Devaiah, V.; Reddy, K. L.; Kumar, M. S. Synthesis and biological activity of fluoroquinolone-pyrrolo[2,1-c][1,4]benzodiazepine conjugates. Bioorg. Med. Chem. 2005, 13, 2021-2029. [2] Valery, N. C.; Nataliya, N. M.; Fedor, V. A.; Svetlana, K. K.; Emiliya, V. N.; Marina, A. E.; Mikhail, I. K.; Marionella, A. K. Synthesis and antimycobacterial evaluation of new (2-oxo-2H-chromen-3-yl) substituted fluoroquinolones. J. Fluorine Chem. 2018, 208, 15-23.”

[0043] Example 2, Preparation of intermediate VI

[0044]

[0045] Intermediate VI was prepared according to the method described in the reference “Liu Zhonghua, Liu Fang, Xue Deming, Li Tao, Gao Liuzhou, Xie Yuluo, Feng Yanfei, Yan Qiang, Wu Shumin, Ni Lili, Hu Guoqiang. Chiral 7-(piperazine-substituted pyrazole aldehyde condensation isoniazid hydrazone) fluoroquinolone carboxylic acid derivatives and their preparation method and application. Chinese invention patent, application publication number: CN104402902A.”

[0046] Example 3, Preparation of compound I-1-1:

[0047]

[0048] Intermediate IV-1 (100 mg, 0.36 mmol), 1H-pyrazole-3-carboxaldehyde (42 mg, 0.44 mmol) and piperidine (6 mg, 0.07 mmol) were reacted in ethanol (8 mL) at 78 °C for 12 hours. After the reaction was completed, it was cooled to room temperature, water (10 mL) was added and stirred constantly, and the obtained solid was filtered and recrystallized to obtain 113 mg of white solid I-1-1, yield: 87.1%, melting point: 288.8-290.4 °C; 1H NMR (600 MHz, 25 °C, DMSO-d6) δ 14.14 (s, 1H), 9.10 (s, 1H), 8.29 (d, J = 6.1 Hz, 1H), 8.13 (d, J = 10.5 Hz, 1H), 8.02 (s, 1H), 8.00 (d, J = 2.4 Hz, 1H), 7.14 (d, J = 2.4 Hz, 1H), 4.70 (q, J = 7.1 Hz, 2H), 1.47 (t, J = 7.1 Hz, 3H) ppm.

[0049] Example 4, Preparation of compound I-1-2:

[0050]

[0051] Intermediate IV-1 (100 mg, 0.36 mmol), 1-methyl-1H-pyrazole-3-carbaldehyde (48 mg, 0.44 mmol) and piperidine (6 mg, 0.07 mmol) were reacted in ethanol (8 mL) at 78 °C for 12 hours. After the reaction was completed, it was cooled to room temperature, water (10 mL) was added and stirred constantly, the obtained solid was filtered and recrystallized to obtain 121 mg of white solid I-1-2, yield: 85.5%, melting point: >300 °C; 1 H NMR (600 MHz, 25 °C, DMSO-d6 + CF3COOH) δ 9.02 (s, 1H), 8.13 (d, J = 5.5 Hz, 1H), 8.04 (d, J = 9.9 Hz, 1H), 7.78 (s, 1H), 7.68 (d, J = 2.8 Hz, 1H), 7.27 (d, J = 2.8 Hz, 1H), 4.55 (q, J = 7.3 Hz, 2H), 3.84 (s, 3H), 1.33 (t, J = 7.3 Hz, 3H) ppm.

[0052] Example 5, Preparation of compound I-2-1:

[0053]

[0054] Intermediate IV-1 (100 mg, 0.36 mmol), 1H-pyrazole-4-carbaldehyde (42 mg, 0.44 mmol) and piperidine (6 mg, 0.07 mmol) were reacted in ethanol (8 mL) at 78 °C for 12 hours. After the reaction was completed, it was cooled to room temperature, water (10 mL) was added and stirred constantly, the obtained solid was filtered and recrystallized to obtain 118 mg of yellow solid I-2-1, yield: 85.1%, melting point: >300 °C; 1HNMR (400 MHz, 25 °C, DMSO-d6) δ 14.21 (s, 2H), 9.09 (s, 1H), 8.35 (s, 2H), 8.19 (d, J = 6.3 Hz, 1H), 8.11 (d, J = 10.8 Hz, 1H), 8.02 (s, 1H), 4.67 (t, J = 7.2 Hz, 2H), 1.47 (t, J = 7.1 Hz, 3H) ppm.

[0055] Example 6, Preparation of compound I-2-2:

[0056]

[0057] Intermediate IV-1 (100 mg, 0.36 mmol), 1 -methyl- lH-pyrazole-4-carboxaldehyde (48 mg, 0.44 mmol) and piperidine (6 mg, 0.07 mmol) were reacted in ethanol (8 mL) at 78 °C for 12 hours. After the reaction was completed, it was cooled to room temperature, water (10 mL) was added and stirred constantly, the obtained solid was filtered and recrystallized to obtain 118 mg of yellow solid I-2-2, yield: 88.1%, melting point: 271.6-273.4 °C; 1 H NMR (600 MHz, 25 °C, DMSO-d6 + CF3COOH) δ 9.01 (s, 1H), 8.49 (s, 1H), 8.37 (s, 1H), 8.03 (dd, J = 7.9, 5.5 Hz, 2H), 7.70 (s, 1H), 4.53 (q, J = 7.3 Hz, 2H), 3.91 (s, 3H), 1.33 (t, J = 7.3 Hz, 3H) ppm.

[0058] Example 7, Preparation of compound I-2-3:

[0059]

[0060] Intermediate IV-1 (100 mg, 0.36 mmol), 1 -methyl- lH-pyrazole-4-carboxaldehyde (48 mg, 0.44 mmol) and piperidine (6 mg, 0.07 mmol) were reacted in ethanol (8 mL) at 78 °C for 12 hours. After the reaction was completed, it was cooled to room temperature, water (10 mL) was added and stirred constantly, the obtained solid was filtered and recrystallized to obtain 118 mg of yellow solid I-2-2, yield: 88.1%, melting point: 271.6-273.4 °C; 1H NMR (400 MHz, 25 °C, DMSO-d6) δ 14.90 (s, 1H), 13.36 (s, 1H), 9.09 (s, 1H), 8.34 (s, 1H), 8.21 (d, J = 6.3 Hz, 1H), 8.10 (d, J = 10.6 Hz, 1H), 7.79 (s, 1H), 4.69 (q, J = 7.1 Hz, 2H), 2.43 (s, 3H), 1.47 (t, J = 7.1 Hz, 3H) ppm.

[0061] Example 8, Preparation of compound I-2-4:

[0062]

[0063] Intermediate IV-1 (100 mg, 0.36 mmol), 3-phenyl-lH-pyrazole-4-carboxaldehyde (75 mg, 0.44 mmol) and piperidine (6 mg, 0.07 mmol) were reacted in ethanol (8 mL) at 78 °C for 12 hours. After the reaction was completed, it was cooled to room temperature, water (10 mL) was added and stirred constantly, the obtained solid was filtered and recrystallized to obtain 141 mg of yellow solid I-2-4, yield: 82.8%, melting point: >300 °C; 1 H NMR (600 MHz, DMSO-d6) δ 14.31 (s, 2H), 9.08 (s, 1H), 8.62 (s, 1H), 8.22 (d, J = 6.2 Hz, 1H), 8.09 (d, J = 10.7 Hz, 1H), 7.78 (s, 1H), 7.68 - 7.66 (m, 2H), 7.53 (q, J = 10.0, 8.7 Hz, 3H), 4.66 (t, J = 7.2 Hz, 2H), 1.42 (t, J = 7.1 Hz, 3H) ppm.

[0064] Example 9, Preparation of compound I-2-5:

[0065]

[0066] Intermediate IV-1 (100 mg, 0.36 mmol), 3-phenyl-lH-pyrazole-4-carboxaldehyde (75 mg, 0.44 mmol) and piperidine (6 mg, 0.07 mmol) were reacted in ethanol (8 mL) at 78 °C for 12 hours. After the reaction was completed, it was cooled to room temperature, water (10 mL) was added and stirred constantly, the obtained solid was filtered and recrystallized to obtain 141 mg of yellow solid I-2-4, yield: 82.8%, melting point: >300 °C; 1H NMR (600 MHz, 25 °C, DMSO-d6) δ 14.90 (s, 1H), 9.10 (s, 1H), 8.54 (s, 1H), 8.22 (d, J = 6.1 Hz, 1H), 8.13 (d, J = 10.5 Hz, 1H), 7.75 (s, 1H), 4.69 (d, J = 7.2 Hz, 2H), 3.92 (s, 3H), 2.32 (s, 3H), 1.46 (t, J = 7.1 Hz, 3H) ppm.

[0067] Example 10, Preparation of compound I-2-6:

[0068]

[0069] Intermediate IV-1 (100 mg, 0.36 mmol), 1,3,5-trimethyl-lH-pyrazole-4-carboxaldehyde (60 mg, 0.44 mmol) and piperidine (6 mg, 0.07 mmol) were reacted in ethanol (8 mL) at 78 °C for 12 hours. After the reaction was completed, it was cooled to room temperature, water (10 mL) was added and stirred constantly, the obtained solid was filtered and recrystallized to obtain 111 mg of yellow solid I-2-6, yield: 77.1%, melting point: 288.5-289.7 °C; 1 H NMR (600 MHz, 25 °C, DMSO-d6) δ 14.88 (s, 1H), 9.10 (s, 1H), 8.18 (d, J = 6.1 Hz, 1H), 8.13 (d, J = 10.5 Hz, 1H), 7.86 (d, J = 1.5 Hz, 1H), 4.68 (t, J = 7.2 Hz, 2H), 3.76 (s, 3H), 2.38 (s, 3H), 2.27 (s, 3H), 1.47 (t, J = 7.1 Hz, 3H) ppm.

[0070] Example 11, Preparation of compound I-2-7:

[0071]

[0072] Intermediate IV-1 (100 mg, 0.36 mmol), 5-chloro-l,3-dimethyl-lH-pyrazole-4-carboxaldehyde (69 mg, 0.44 mmol) and piperidine (6 mg, 0.07 mmol) were reacted in ethanol (8 mL) at 78 °C for 12 hours. After the reaction was completed, it was cooled to room temperature, water (10 mL) was added and stirred constantly, the obtained solid was filtered and recrystallized to obtain 113 mg of white solid I-2-7, yield: 73.4%, melting point: 279.1-281.5; 1H NMR (400 MHz, DMSO-d6) δ 14.86 (s, 1H), 9.12 (s, 1H), 8.23 (d, J = 6.2 Hz, 1H), 8.16 (d, J = 10.6 Hz, 1H), 7.79 (s, 1H), 4.69 (d, J = 7.2 Hz, 2H), 3.85 (s, 3H), 2.34 (s, 3H), 1.46 (s, 3H) ppm.

[0073] Example 12, Preparation of compound I-3:

[0074]

[0075] Intermediate IV-1 (100 mg, 0.36 mmol), 1-methyl-1H-pyrazole-5-carbaldehyde (48 mg, 0.44 mmol) and piperidine (6 mg, 0.07 mmol) were reacted in ethanol (8 mL) at 78 °C for 12 hours. After the reaction was completed, it was cooled to room temperature, water (10 mL) was added and stirred constantly, and the obtained solid was filtered and recrystallized to obtain 92 mg of yellow solid I-3, yield: 68.7%, melting point: 288.4-290.6 °C; 1 H NMR (600 MHz, 25 °C, DMSO-d6) δ 14.83 (s, 1H), 9.12 (s, 1H), 8.36 (d, J = 5.7 Hz, 1H), 8.18 (d, J = 10.4 Hz, 1H), 8.03 (s, 1H), 7.70 (s, 1H), 7.28 (s, 1H), 4.69 (d, J = 7.2 Hz, 2H), 4.05 (s, 3H), 1.47 (d, J = 7.3 Hz, 3H) ppm.

[0076] Example 13, Preparation of compound I-4-1:

[0077]

[0078] Intermediate IV-2 (80 mg, 0.26 mmol), 1,3-dimethylpyrazole-4-carbaldehyde (39 mg, 0.32 mmol) and piperidine (5 mg, 0.05 mmol) were reacted in ethanol (8 mL) at 78 °C for 24 hours. After the reaction was completed, it was cooled to room temperature, water (10 mL) was added and stirred constantly, and the obtained solid was filtered and recrystallized to obtain 67 mg of yellow solid I-4-1, yield: 62.0%, melting point: 282.3-284.4 °C; 1H NMR (600 MHz, 25 °C, DMSO-d6 + CF3COOH) δ 8.97 (s, 1H), 8.66 (s, 1H), 8.03 - 8.01 (m, 2H), 7.63 (s, 1H), 4.46 (t, J = 7.7 Hz, 2H), 3.86 (s, 3H), 2.21 (s, 3H), 1.66 (t, J = 7.7 Hz, 2H), 1.10 (q, J = 7.5 Hz, 2H), 0.59 (t, J = 7.4 Hz, 3H) ppm.

[0079] Example 14, Preparation of compound I-4-2:

[0080]

[0081] Intermediate IV-3 (80 mg, 0.28 mmol), 1,3-dimethylpyrazole-4-carboxaldehyde (42 mg, 0.34 mmol) and piperidine (5 mg, 0.05 mmol) were reacted in ethanol (8 mL) at 78 °C for 12 hours. After the reaction was completed, it was cooled to room temperature, water (10 mL) was added and stirred constantly, the obtained solid was filtered and recrystallized to obtain 75 mg of yellow solid I-4-2 with a yield of 68.2%, melting point: 260.2-262.7 °C; 1 H NMR (600 MHz, 25 °C, DMSO-d6) δ 14.80 (s, 1H), 9.10 (s, 1H), 8.52 (s, 1H), 8.13 (d, J = 6.1 Hz, 1H), 8.09 (d, J = 10.8 Hz, 1H), 7.68 (s, 1H), 6.12 (ddd, J = 16.8, 10.5, 5.2 Hz, 1H), 5.39 - 5.33 (m, 4H), 3.92 (s, 3H), 2.31 (s, 3H) ppm.

[0082] Example 15, Preparation of compound I-4-3:

[0083]

[0084] Intermediate IV-4 (80 mg, 0.28 mmol), 1,3-dimethylpyrazole-4-carboxaldehyde (42 mg, 0.34 mmol) and piperidine (5 mg, 0.05 mmol) were reacted in ethanol (8 mL) at 78 °C for 12 hours. After the reaction was completed, it was cooled to room temperature, water (10 mL) was added and stirred constantly, the obtained solid was filtered and recrystallized to obtain 75 mg of yellow solid I-4-2 with a yield of 68.2%, melting point: 260.2-262.7 °C; 1H NMR (600 MHz, 25 °C, DMSO-d6 + CF3COOH) δ 8.94 (s, 1H), 8.60 (s, 1H), 8.45 (d, J = 5.9 Hz, 1H), 7.93 (d, J = 10.4 Hz, 1H), 7.60 (s, 1H), 3.80 (s, 3H), 3.73 (dt, J = 7.1, 3.3 Hz, 1H), 2.16 (s, 3H), 1.20 (d, J = 7.2 Hz, 2H), 0.97 (d, J = 4.2 Hz, 2H) ppm.

[0085] Example 16, Preparation of compound I-4-4:

[0086]

[0087] Intermediate IV-5 (90 mg, 0.30 mmol), 1,3-dimethylpyrazole-4-carboxaldehyde (47 mg, 0.36 mmol) and piperidine (5 mg, 0.05 mmol) were reacted in ethanol (8 mL) at 78 °C for 24 h. After the reaction was completed, it was cooled to room temperature, water (10 mL) was added and stirred constantly, the obtained solid was filtered and recrystallized to obtain 80 mg of yellow solid I-4-4 with a yield of 76.9%, melting point: 274.1-276.2 °C; 1 H NMR (600 MHz, 25 °C, DMSO-d6 + CF3COOH) δ 9.26 (s, 1H), 8.67 (s, 1H), 8.13 (d, J = 5.7 Hz, 1H), 8.05 (d, J = 10.2 Hz, 1H), 7.62 (s, 1H), 4.32 (d, J = 7.3 Hz, 2H), 3.88 (s, 3H), 2.23 (s, 3H), 1.08 (td, J = 7.7, 3.9 Hz, 1H), 0.57 (d, J = 7.4 Hz, 2H), 0.26 (d, J = 5.1 Hz, 2H) ppm.

[0088] Example 17, Preparation of compound I-4-5:

[0089]

[0090] Intermediate IV-6 (90 mg, 0.27 mmol), 1,3-dimethylpyrazole-4-carboxaldehyde (41 mg, 0.33 mmol) and piperidine (5 mg, 0.05 mmol) were reacted in ethanol (8 mL) at 78 °C for 24 h. After the reaction was completed, it was cooled to room temperature, water (10 mL) was added and stirred constantly, the obtained solid was filtered and recrystallized to obtain 81 mg of yellow solid I-4-5 with a yield of 68.1%, melting point: 275.6-277.9 °C; 1H NMR (600 MHz, 25 °C, DMSO-d6) δ 14.88 (s, 1H), 9.10 (s, 1H), 8.54 (s, 1H), 8.25 (d, J = 6.1 Hz, 1H), 8.12 (d, J = 10.4 Hz, 1H), 7.73 (s, 1H), 4.64 (d, J = 7.6 Hz, 2H), 3.92 (s, 3H), 2.46 (q, J = 7.8 Hz, 1H), 2.31 (s, 3H), 1.66 (ddd, J = 18.4, 10.8, 6.7 Hz, 4H), 1.50 (q, J = 6.3, 5.7 Hz, 2H), 1.33 - 1.28 (m, 2H) ppm.

[0091] Example 18, Preparation of compound I-4-6:

[0092]

[0093] Intermediate IV-7 (80 mg, 0.24 mmol), 1,3-dimethylpyrazole-4-carboxaldehyde (35 mg, 0.29 mmol) and piperidine (5 mg, 0.05 mmol) were reacted in ethanol (8 mL) at 78 °C for 12 hours. After the reaction was completed, it was cooled to room temperature, water (10 mL) was added and stirred constantly, the obtained solid was filtered and recrystallized to obtain 82 mg of yellow solid I-4-6, yield 78.1%, melting point: >300 °C; 1 H NMR (600 MHz, DMSO-d6) δ 14.81 (s, 1H), 9.35 (s, 1H), 8.51 (s, 1H), 8.11 (d, J = 10.3 Hz, 2H), 7.44 - 7.36 (m, 6H), 5.95 (s, 2H), 3.90 (s, 3H), 2.26 (s, 3H) ppm.

[0094] Example 19, Preparation of compound I-4-7:

[0095]

[0096] Intermediate IV-8 (80 mg, 0.23 mmol), 1,3-dimethylpyrazole-4-carboxaldehyde (34 mg, 0.27 mmol) and piperidine (5 mg, 0.05 mmol) were reacted in ethanol (8 mL) at 78 °C for 12 hours. After the reaction was completed, it was cooled to room temperature, water (10 mL) was added and stirred constantly, the obtained solid was filtered and recrystallized to obtain 84 mg of yellow solid I-4-7, yield 80.8%, melting point: >300 °C; 1H NMR (600 MHz, 25 °C, DMSO-d6) δ 14.73 (s, 1H), 9.30 (s, 1H), 8.52 (s, 1H), 8.12 (d, J = 10.8 Hz, 1H), 8.08 (d, J = 6.2 Hz, 1H), 7.54 (s, 1H), 7.44 - 7.40 (m, 2H), 7.28 (dd, J = 10.8, 8.4 Hz, 1H), 7.20 (td, J = 7.6, 1.2 Hz, 1H), 6.02 (s, 2H), 3.91 (s, 3H), 2.27 (s, 3H) ppm.

[0097] Example 20, Preparation of compound I-4-8:

[0098]

[0099] Intermediate IV-9 (80 mg, 0.23 mmol), 1,3-dimethylpyrazole-4-carboxaldehyde (34 mg, 0.27 mmol) and piperidine (5 mg, 0.05 mmol) were reacted in ethanol (8 mL) at 78 °C for 12 hours. After the reaction was completed, it was cooled to room temperature, water (10 mL) was added and stirred constantly, the obtained solid was filtered and recrystallized to obtain 80 mg of yellow solid I-4-8 with a yield of 76.9%, melting point: 281.6-283.1 °C; 1 H NMR (600 MHz, 25 °C, DMSO-d6) δ 14.73 (s, 1H), 9.30 (s, 1H), 8.52 (s, 1H), 8.12 (d, J = 10.8 Hz, 1H), 8.08 (d, J = 6.2 Hz, 1H), 7.54 (s, 1H), 7.44 - 7.40 (m, 2H), 7.28 (dd, J = 10.8, 8.4 Hz, 1H), 7.20 (td, J = 7.6, 1.2 Hz, 1H), 6.02 (s, 2H), 3.91 (s, 3H), 2.27 (s, 3H) ppm.

[0100] Example 21, Preparation of compound I-4-9:

[0101]

[0102] Intermediate IV-10 (80 mg, 0.23 mmol), 1,3-dimethylpyrazole-4-carboxaldehyde (34 mg, 0.27 mmol) and piperidine (5 mg, 0.05 mmol) were reacted in ethanol (8 mL) at 78 °C for 12 hours. After the reaction was completed, it was cooled to room temperature, water (10 mL) was added and stirred constantly, the obtained solid was filtered and recrystallized to obtain 80 mg of yellow solid I-4-9 with a yield of 76.9%, melting point: >300 °C; 1H NMR (600 MHz, DMSO-d6) δ 14.76 (s, 1H), 9.38 (s, 1H), 8.49 (s, 1H), 8.11 (d, J = 10.7 Hz, 1H), 7.99 (d, J = 6.1 Hz, 1H), 7.74 (d, J = 8.0 Hz, 2H), 7.58 (d, J = 8.0 Hz, 2H), 7.34 (s, 1H), 6.06 (s, 2H), 3.90 (s, 3H), 2.20 (s, 3H) ppm.

[0103] Example 22, Preparation of compound I-4-10:

[0104]

[0105] Intermediate IV-11 (100 mg, 0.25 mmol), 1,3-dimethylpyrazole-4-carboxaldehyde (37 mg, 0.30 mmol) and piperidine (4 mg, 0.04 mmol) were reacted in ethanol (8 mL) at 78 °C for 8 hours. After the reaction was completed, it was cooled to room temperature, water (10 mL) was added and stirred constantly, and the obtained solid was filtered and recrystallized to obtain 68 mg of yellow solid I-4-10 with a yield of 54.0%, melting point: >300 °C; 1 H NMR (600 MHz, DMSO-d6) δ 14.76 (s, 1H), 9.38 (s, 1H), 8.49 (s, 1H), 8.11 (d, J = 10.7 Hz, 1H), 7.99 (d, J = 6.1 Hz, 1H), 7.74 (d, J = 8.0 Hz, 2H), 7.58 (d, J = 8.0 Hz, 2H), 7.34 (s, 1H), 6.06 (s, 2H), 3.90 (s, 3H), 2.20 (s, 3H) ppm.

[0106] Example 23, Preparation of compound I-4-11:

[0107]

[0108] Intermediate IV-12 (80 mg, 0.23 mmol), 1,3-dimethylpyrazole-4-carboxaldehyde (34 mg, 0.27 mmol) and piperidine (4 mg, 0.04 mmol) were reacted in ethanol (8 mL) at 78 °C for 8 hours. After the reaction was completed, it was cooled to room temperature, water (10 mL) was added and stirred constantly, and the obtained solid was filtered and recrystallized to obtain 80 mg of yellow solid I-4-11 with a yield of 76.9%, melting point: >300 °C; 1H NMR (600 MHz, 25 °C, DMSO-d6) δ 14.81 (s, 1H), 9.33 (s, 1H), 8.51 (s, 1H), 8.11 - 8.08 (m, 2H), 7.42 (s, 1H), 7.28 (d, J = 7.9 Hz, 2H), 7.18 (d, J = 7.9 Hz, 2H), 5.89 (s, 2H), 3.91 (s, 3H), 2.27 (s, 3H), 2.26 (s, 3H) ppm.

[0109] Example 24, Preparation of compound I-4-12:

[0110]

[0111] Intermediate IV-13 (100 mg, 0.27 mmol), 1,3-dimethylpyrazole-4-carboxaldehyde (40 mg, 0.32 mmol) and piperidine (5 mg, 0.05 mmol) were reacted in ethanol (8 mL) at 78 °C for 8 hours. After the reaction was completed, it was cooled to room temperature, water (10 mL) was added and stirred constantly, and the obtained solid was filtered and recrystallized to obtain 106 mg of yellow solid I-4-12 with a yield of 82.2%, melting point: >300 °C; 1 H NMR (600 MHz, 25 °C, DMSO-d6) δ 14.72 (s, 1H), 9.29 (s, 1H), 8.52 (s, 1H), 8.13 (d, J = 10.7 Hz, 1H), 8.07 (d, J = 6.1 Hz, 1H), 7.55 (s, 1H), 7.49 (td, J = 8.7, 6.3 Hz, 1H), 7.35 (td, J = 8.9, 4.5 Hz, 1H), 7.08 (td, J = 8.5, 2.6 Hz, 1H), 5.98 (s, 2H), 3.91 (s, 3H), 2.27 (s, 3H) ppm.

[0112] Example 25, Preparation of compound I-4-13:

[0113]

[0114] Intermediate IV-14 (80 mg, 0.20 mmol), 1,3-dimethylpyrazole-4-carboxaldehyde (29 mg, 0.24 mmol) and piperidine (4 mg, 0.04 mmol) were reacted in ethanol (8 mL) at 78 °C for 8 hours. After the reaction was completed, it was cooled to room temperature, water (10 mL) was added and stirred constantly, and the obtained solid was filtered and recrystallized to obtain 81 mg of yellow solid I-4-13 with a yield of 80.2%, melting point: >300 °C; 1H NMR (600 MHz, 25 °C, DMSO-d6) δ 14.70 (s, 1H), 9.26 (s, 1H), 8.49 (s, 1H), 8.14 (d, J = 10.7 Hz, 1H), 7.78 - 7.76 (m, 2H), 7.50 (s, 1H), 7.35 (dd, J = 8.4, 2.1 Hz, 1H), 7.08 (d, J = 8.4 Hz, 1H), 5.98 (s, 2H), 3.89 (s, 3H), 2.22 (s, 3H) ppm.

[0115] Example 26, Preparation of compound I-4-14:

[0116]

[0117] Intermediate IV-15 (100 mg, 0.27 mmol), 1,3-dimethylpyrazole-4-carboxaldehyde (40 mg, 0.32 mmol) and piperidine (5 mg, 0.05 mmol) were reacted in ethanol (8 mL) at 78 °C for 8 hours. After the reaction was completed, it was cooled to room temperature, water (10 mL) was added and stirred constantly, and the obtained solid was filtered and recrystallized to obtain 95 mg of yellow solid I-4-14 with a yield of 73.6%, melting point: >300 °C; 1 H NMR (600 MHz, 25 °C, DMSO-d6) δ 14.70 (s, 1H), 9.26 (s, 1H), 8.49 (s, 1H), 8.14 (d, J = 10.7 Hz, 1H), 7.78 - 7.76 (m, 2H), 7.50 (s, 1H), 7.35 (dd, J = 8.4, 2.1 Hz, 1H), 7.08 (d, J = 8.4 Hz, 1H), 5.98 (s, 2H), 3.89 (s, 3H), 2.22 (s, 3H) ppm.

[0118] Example 27, Preparation of compound I-4-15:

[0119]

[0120] Intermediate IV-16 (100 mg, 0.27 mmol), 1,3-dimethylpyrazole-4-carboxaldehyde (40 mg, 0.27 mmol) and piperidine (5 mg, 0.05 mmol) were reacted in ethanol (8 mL) at 78 °C for 12 hours. After the reaction was completed, it was cooled to room temperature, water (10 mL) was added and stirred constantly, and the obtained solid was filtered and recrystallized to obtain 116 mg of yellow solid I-4-15 with a yield of 89.9%, melting point: >300 °C; 1H NMR (600 MHz, 25 °C, DMSO-d6) δ 14.81 (s, 1H), 9.31 (s, 1H), 8.49 (d, J = 4.3 Hz, 2H), 8.11 - 8.08 (m, 2H), 7.88 - 7.85 (m, 1H), 7.68 (d, J = 7.9 Hz, 1H), 7.45 (s, 1H), 7.35 (dd, J = 7.4, 4.9 Hz, 1H), 6.06 (s, 2H), 3.90 (s, 3H), 2.26 (s, 3H) ppm.

[0121] Example 28, Preparation of compound I-4-16:

[0122]

[0123] Intermediate IV-17 (80 mg, 0.24 mmol), 1,3-dimethylpyrazole-4-carboxaldehyde (35 mg, 0.28 mmol) and piperidine (4 mg, 0.04 mmol) were reacted in ethanol (8 mL) at 78 °C for 12 h. After the reaction was completed, it was cooled to room temperature, water (10 mL) was added and stirred constantly, and the obtained solid was filtered and recrystallized to obtain 87 mg of yellow solid I-4-16 with a yield of 82.9%, melting point: >300 °C; 1 H NMR (600 MHz, 25 °C, DMSO-d6) δ 14.81 (s, 1H), 9.31 (s, 1H), 8.49 (d, J = 4.3 Hz, 2H), 8.11 - 8.08 (m, 2H), 7.88 - 7.85 (m, 1H), 7.68 (d, J = 7.9 Hz, 1H), 7.45 (s, 1H), 7.35 (dd, J = 7.4, 4.9 Hz, 1H), 6.06 (s, 2H), 3.90 (s, 3H), 2.26 (s, 3H) ppm.

[0124] Example 29, Preparation of compound II:

[0125]

[0126] Intermediate V (100 mg, 0.33 mmol), 1,3-dimethylpyrazole-4-carboxaldehyde (49 mg, 0.40 mmol) and piperidine (5 mg, 0.05 mmol) were reacted in ethanol (8 mL) at 78 °C for 12 h. After the reaction was completed, it was cooled to room temperature, water (10 mL) was added and stirred constantly, and the obtained solid was filtered and recrystallized to obtain 85 mg of yellow solid II with a yield of 63.0%, melting point: >300 °C; 1H NMR (400 MHz, 25 °C, DMSO-d6) δ 14.89 (s, 1H), 9.10 (s, 1H), 8.52 (s, 1H), 7.72 (d, J = 9.9 Hz, 1H), 7.59 (s, 1H), 5.02 (dt, J = 6.9, 2.2 Hz, 1H), 4.69 (dd, J = 11.6, 1.8 Hz, 1H), 4.52 (dd, J = 11.5, 2.5 Hz, 1H), 3.91 (s, 3H), 2.25 (s, 3H), 1.50 (d, J = 6.7 Hz, 3H) ppm.

[0127] Example 30, Preparation of compound III-1 :

[0128]

[0129] Intermediate VI (100 mg, 0.34 mmol), 1-methyl-1H-pyrazole-3-carboxaldehyde (45 mg, 0.41 mmol) and acetic acid (4 mg, 0.07 mmol) were reacted in ethanol (8 mL) at 78 °C for 2 hours. After the reaction was completed, it was cooled to room temperature, water (10 mL) was added and stirred constantly, the obtained solid was filtered and recrystallized to obtain 94 mg of yellow solid III-1 with a yield of 71.8%, melting point: >300 °C; 1 H NMR (600 MHz, DMSO-d6) δ 15.36 (s, 1H), 10.30 (s, 1H), 8.95 (s, 1H), 8.28 (d, J = 3.5 Hz, 1H), 7.70 (d, J = 2.3 Hz, 1H), 7.62 (d, J = 12.4 Hz, 1H), 6.46 (d, J = 2.3 Hz, 1H), 4.96 (d, J = 7.1 Hz, 1H), 4.63 (d, J = 11.4 Hz, 1H), 4.44 (d, J = 11.2 Hz, 1H), 3.85 (s, 3H), 1.48 (d, J = 6.9 Hz, 3H) ppm.

[0130] Example 31, Preparation of compound III-2:

[0131]

[0132] Intermediate VI (100 mg, 0.34 mmol), 1-methyl-1H-pyrazole-3-carboxaldehyde (45 mg, 0.41 mmol) and acetic acid (4 mg, 0.07 mmol) were reacted in ethanol (8 mL) at 78 °C for 2 hours. After the reaction was completed, it was cooled to room temperature, water (10 mL) was added and stirred constantly, the obtained solid was filtered and recrystallized to obtain 94 mg of yellow solid III-1 with a yield of 71.8%, melting point: >300 °C; 1H NMR (600 MHz, 25 °C, DMSO-d6) δ 15.42 (s, 1H), 9.94 (s, 1H), 8.93 (s, 1H), 8.24 (d, J = 3.2 Hz, 1H), 7.86 (s, 1H), 7.59 (d, J = 12.5 Hz, 1H), 4.95 (t, J = 6.7 Hz, 1H), 4.61 (dd, J = 11.4, 1.9 Hz, 1H), 4.42 (dd, J = 11.5, 2.4 Hz, 1H), 3.77 (s, 3H), 2.32 (s, 3H), 1.48 (d, J = 6.8 Hz, 3H) ppm.

[0133] Example 32, in vitro antimicrobial activity of pyrazole cyano vinyl pyridone acid compounds

[0134] The minimum inhibitory concentration (MIC) of the pyrazole cyano vinyl pyridone acid compounds prepared in Examples 3-31 against gram-positive bacteria (methicillin-resistant Staphylococcus aureus ATCC 43300, Staphylococcus aureus ATCC 12600, Staphylococcus aureus ATCC 25923, Staphylococcus aureus ATCC 29213, Enterococcus faecalis p1-2007226001, Streptococcus pyogenes ATCC 19615) and gram-negative bacteria (Klebsiella pneumoniae CMCC (B) 46117, Escherichia coli ATCC 8739, Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 15442, Pseudomonas aeruginosa ATCC 27853, Acinetobacter baumannii ATCC 19606) was determined by the method recommended by the Clinical and Laboratory Standards Institute (CLSI) using a two-fold dilution method. The test compound was dissolved in a small amount of dimethyl sulfoxide and diluted with culture solution to 64 μg / mL. After continuous dilution, the microorganism was inoculated and incubated at 37 °C for 24 hours. After the culture plate was placed on a shaker and shaken well, the MIC was determined at a wavelength of 600 nm and read by the naked eye. The results are shown in Tables 1 and 2.

[0135] Table 1, in vitro activity of the pyrazole cyano vinyl pyridone acid compounds prepared in Examples 3-31 against gram-positive bacteria (MIC, μg / mL)

[0136]

[0137]

[0138] As shown in Table 1, the pyrazole cyanovinylpyridone acid compounds I-III prepared in the present application exhibit good inhibitory effect on the tested gram-positive bacteria, and the dimethylpyrazole derivatives have relatively better inhibitory activity. In particular, the ofloxacin derivative II exhibits very good inhibitory activity on the five tested gram-positive bacteria, with the MIC value of 0.0625-0.5 μg / mL, which is much better than the reference drugs norfloxacin, ciprofloxacin and levofloxacin. When the connecting segment is changed from cyanovinyl to hydrazone, the activity is greatly decreased, which indicates the advantage of cyanovinyl structure in improving the antibacterial activity.

[0139] Table 2, in vitro activity of the pyrazole cyanovinylpyridone acid compounds prepared in Examples 3-31 against gram-negative bacteria (MIC, μg / mL)

[0140]

[0141] As shown in Table 2, the pyrazole cyanovinylpyridone acid compounds I-III prepared in the present application exhibit moderate to good inhibitory effect on the tested gram-negative bacteria, and the phenylpyrazole derivatives and dimethylpyrazole derivatives have relatively better inhibitory activity. Among them, the 2,4-dichlorobenzyl compounds I-4-13 and the ofloxacin derivative II exhibit excellent antibacterial activity against the tested Klebsiella pneumoniae, Escherichia coli and Acinetobacter baumannii, with the MIC value of 0.0625-0.25 μg / mL, which is better than the three reference drugs. In addition, when the connecting segment is changed to hydrazone, the antibacterial activity is obviously decreased.

[0142] Example 33, pharmaceutical use of the pyrazole cyanovinylpyridone acid compounds

[0143] According to the above antimicrobial activity test results, the pyrazole cyanovinylpyridone acid compounds of the present application have good antibacterial activity, and can be made into antibacterial drugs for clinical use. These drugs can be single formulations, such as those prepared from one kind of pyrazole cyanovinylpyridone acid compound and pharmaceutically acceptable excipients; or can be compound formulations, such as those prepared from one kind of pyrazole cyanovinylpyridone acid compound, an antibacterial active ingredient (such as norfloxacin, etc.) and pharmaceutically acceptable excipients, or those prepared from several pyrazole cyanovinylpyridone acid compounds of different structures and pharmaceutically acceptable excipients. The formulation types include but are not limited to tablet, capsule, powder, granule, drop pill, injection, powder injection, solution, suspension, emulsion, suppository, ointment, gel, film, aerosol, transdermal absorption patch, etc., as well as various sustained-release, controlled-release formulations and nano-preparations.

[0144] 1. Preparation of compound II tablet

[0145] Prescription: Compound II 10g, corn starch 50g, lactose 187g, magnesium stearate 3.0g, 70% ethanol solution, q.s. 1000 tablets.

[0146] Method of preparation: corn starch is dried at 105°C for 5 hours; Compound II is mixed with lactose and corn starch, and then the mixture is made into soft material with 70% ethanol solution, and then wet granules are prepared by sieving. Magnesium stearate is added, and then the mixture is tableted to obtain tablets, each weighing 250mg, and containing 10mg of active ingredient.

[0147] 2. Preparation of Compound II Capsules

[0148] Prescription: Compound II 25g, modified starch (120 mesh) 12.5g, microcrystalline cellulose (100 mesh) 7.5g, low-substituted hydroxypropyl cellulose (100 mesh) 2.5g, talc (100 mesh) 2.0g, sweetening agent 1.25g, orange flavor 0.25g, pigment q.s., water q.s., 1000 capsules.

[0149] Method of preparation: the prescription amount of Compound II is micronized and pulverized into fine powder, which is then mixed with the prescription amount of modified starch, microcrystalline cellulose, low-substituted hydroxypropyl cellulose, talc, sweetening agent, orange flavor and pigment. The mixture is made into soft material with water, and then granulated with a 12-14 mesh sieve. The granules are dried at 40-50°C, sieved, and then filled into empty capsules to obtain the capsules, each weighing 50mg, and containing 25mg of active ingredient.

[0150] 3. Preparation of Compound II Granules

[0151] Prescription: Compound II-1-22 26g, dextrin 120g, sucrose 280g.

[0152] Method of preparation: Compound II, dextrin and sucrose are mixed, and then wet granulation is performed. The granules are dried at 60°C, and then divided into portions.

[0153] 4. Preparation of Compound II Injection

[0154] Prescription: Compound II 10g, propylene glycol 500mL, water for injection 500mL, 1000mL.

[0155] Method of preparation: Compound II is weighed, and then added to propylene glycol and water for injection. After stirring and dissolving, 1g of activated carbon is added. After stirring thoroughly, the mixture is left to stand for 15 minutes. The activated carbon is removed by filtration with a 5μm titanium rod. The filtrate is then filtered with microporous membranes with pore sizes of 0.45μm and 0.22μm, respectively. Finally, the filtrate is filled into 10mL ampoules, and then sterilized by flowing steam at 100°C for 45 minutes to obtain the injection.

[0156] 5. Preparation of Compound II Powder Injection

[0157] Preparation: Compound II sterile powder is sub-packed under sterile condition, and the preparation is completed.

[0158] 6. Preparation of Compound II eye drops

[0159] Prescription: Compound II 3.78 g, sodium chloride 0.9 g, boric acid buffer solution, and distilled water to 1000 mL.

[0160] Preparation: Compound II and sodium chloride are weighed and added to 500 mL of distilled water. After complete dissolution, the pH is adjusted to 6.5 with boric acid buffer solution, and distilled water is added to 1000 mL. After uniform stirring, the mixture is filtered through a microporous filter, filled, sealed, and sterilized by flowing steam at 100°C for 1 hour, and the preparation is completed.

[0161] 7. Preparation of Compound II liniment

[0162] Prescription: Compound II 4 g, potassium soap 7.5 g, camphor 5 g, and distilled water to 100 mL.

[0163] Preparation: Camphor is dissolved in 95% volume percentage ethanol solution and prepared for use. Potassium soap is heated and liquefied and prepared for use. Compound II is weighed, and the potassium soap solution and camphor ethanol solution are added under constant stirring. Distilled water is gradually added until the total amount, and the preparation is completed.

[0164] 8. Preparation of Compound II suppository

[0165] Prescription: Compound II 4 g, gelatin 14 g, glycerol 70 g, and distilled water to 100 mL, and 100 suppositories are prepared.

[0166] Preparation: Gelatin and glycerol are weighed and added to 100 mL of distilled water. When the mixture is heated to 60°C in a water bath and becomes paste-like, Compound II is added, and the mixture is stirred uniformly. When the mixture is near solidification, it is poured into a vaginal suppository mold, cooled, and solidified, and the preparation is completed.

[0167] 9. Preparation of Compound II ointment

[0168] Prescription: Compound II 0.5-2 g, cetyl alcohol 6-8 g, white vaseline 8-10 g, liquid paraffin 8-19 g, monoglyceride 2-5 g, polyoxyethylene (40) stearate 2-5 g, glycerol 5-10 g, and nipagin ethyl ester 0.1 g, and distilled water to 100 g.

[0169] Preparation: Cetyl alcohol, white vaseline, liquid paraffin, monoglyceride, and polyoxyethylene (40) stearate are heated and completely dissolved, mixed uniformly, and kept at 80°C as an oil phase. Nipagin ethyl ester is added to glycerol and distilled water, heated to 85°C to dissolve, and then added to the oil phase under constant stirring. After emulsification, Compound II is added, the mixture is stirred and cooled, and the preparation is completed.

[0170] 10. Preparation of compound II aerosol

[0171] Prescription: compound II 2.5g, Span20 3g, talc (100 mesh) 4g, trichloromonofluoromethane to the appropriate amount.

[0172] Preparation: compound II, Span20 and talc are dried in a vacuum drying oven for several hours, cooled to room temperature in a desiccator, and then crushed into powder by an air flow crusher. The powder is mixed according to the prescription amount, filled into a sealed container, and then trichloromonofluoromethane is added to the specified amount.

[0173] In summary, the present application provides pyrazole cyano vinyl pyridone acid compounds and their pharmaceutically acceptable salts. The present application uses drug design splicing principle. Unlike the traditional quinolone antibacterial drugs using aliphatic heterocyclic modification at C-7 position, the present study uses pyrazole cyano vinyl or pyrazole hydrazone to reconstruct at quinolone C-7 position, and introduces alkyl, alkenyl, aryl or heterocyclic group at quinolone N-1 position to adjust the physicochemical properties and affinity to target. A series of pyrazole cyano vinyl pyridone acid compounds and their pharmaceutically acceptable salts are designed and synthesized. These compounds are found to have good inhibitory activity on gram-positive bacteria (methicillin-resistant Staphylococcus aureus ATCC 43300, Staphylococcus aureus ATCC 12600, Staphylococcus aureus ATCC 25923, Staphylococcus aureus ATCC 29213, Enterococcus faecalis p1-2007226001, Streptococcus pyogenes ATCC 19615) and gram-negative bacteria (Klebsiella pneumoniae CMCC(B)46117, Escherichia coli ATCC 8739, Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 15442, Pseudomonas aeruginosa ATCC 27853, Acinetobacter baumannii ATCC 19606) in vitro, and can be used for preparing antibacterial drugs, providing more efficient and safe candidate drugs for clinical antibacterial treatment, and helping to solve the increasingly serious drug resistance, stubborn pathogenic microorganisms and new harmful microorganisms and other clinical treatment problems.

[0174] Finally, it should be pointed out that the above examples are only used to illustrate the technical solutions of the present application and are not limiting. Although the present application has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solutions of the present application can be modified or replaced by equivalents without departing from the purpose and scope of the present application, and all should be covered in the scope of the claims of the present application.

Claims

1. A pyrazole cyanovinylpyridone acid compound represented by the following formula: ###0001### and pharmaceutically acceptable salts thereof, characterized in that, The structure is shown in general formula I-III: In the formula, n is an integer from 0 to 17; R 1 is alkyl, alkenyl, alkynyl, cycloalkyl and heterocycloalkyl, hydroxy, cyano, ester, carboxyl, phenyl, substituted phenyl or heteroaryl; R 2 , R 3 is hydrogen, halogen or alkyl.

2. The pyrazole cyanoacetylene pyridone acid compound and its pharmaceutically acceptable salt according to claim 1, wherein n is any one of 0, 1 or 3. n is any one of 0, 1 or 3; R 1 is any of alkyl, alkenyl, cycloalkyl, phenyl, substituted phenyl or heteroaryl; R 2 , R 3 is any of hydrogen, halogen or alkyl.

3. The pyrazole cyanovinylpyridone acid compound and pharmaceutically acceptable salt thereof according to Claim 1, wherein The compound is any one of the following compounds:

4. The pyrazole cyanovinylpyridone acid compounds and pharmaceutically acceptable salts thereof according to Claim 1, wherein The pharmaceutically acceptable salt is hydrochloride, bromide, iodide, sulfate, nitrate, trifluoroacetate or acetate.

5. A process for preparing the pyrazole cyanovinylpyridone acid compound and pharmaceutically acceptable salt thereof according to any one of claims 1 to 4, characterized by, The method is as follows: a. adding the compounds shown in general formula IV and V into ethanol, adding various pyrazole aldehyde compounds and refluxing under the action of a base to obtain the pyrazole cyanoacetylene pyridone acid compounds shown in general formula I and II, wherein: n is any one of 0, 1 or 3, R 1 is alkyl, alkenyl, cycloalkyl, phenyl, substituted phenyl or heteroaryl; b. adding the compound shown in general formula VI into ethanol, adding various pyrazole aldehyde compounds and refluxing under the action of an acid to obtain the pyrazole cyanoacetylene pyridone acid compound shown in general formula III; c. preparation of the pharmaceutically acceptable salt of the pyrazole cyanoacetylene pyridone acid compound shown in general formula I-III: dissolving the pyrazole cyanoacetylene pyridone acid compound shown in general formula I-III in an organic solvent, adding a pharmaceutically acceptable acid and reacting until no precipitate is generated to obtain the pharmaceutically acceptable salt of the pyrazole cyanoacetylene pyridone acid compound shown in general formula I-III.

6. The method according to claim 5, wherein In step a, the base is piperidine, the substance amount ratio of the intermediate IV or V, the pyrazole aldehyde compound and the base is 1:1-2:0.2-1, and the reaction is specifically carried out in ethanol at 60-90℃ for 8-24 hours; In step b, the acid is glacial acetic acid, the substance amount ratio of the intermediate VI, the pyrazole aldehyde compound and the acid is 1:1-2:0.2-0.5, and the reaction is specifically carried out in ethanol at 60-90℃ for 1-4 hours; In step c, the organic solvent is at least one of chloroform, acetone, acetonitrile, diethyl ether, tetrahydrofuran, dioxane or N,N-dimethylformamide; and the pharmaceutically acceptable acid is hydrochloric acid or sulfuric acid.

7. Use of the pyrazole cyanoacetylene pyridone acid compound and its pharmaceutically acceptable salt according to any one of claims 1-4 in the preparation of an antibacterial drug.

8. Use according to claim 7, wherein the compound is ###0002### The bacteria are any one or more of methicillin-resistant Staphylococcus aureus ATCC 43300, Staphylococcus aureus ATCC 12600, Staphylococcus aureus ATCC 25923, Staphylococcus aureus ATCC 29213, Enterococcus faecalis p1-2007226001, Klebsiella pneumoniae CMCC(B)46117, Escherichia coli ATCC 8739, Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 15442, Pseudomonas aeruginosa ATCC 27853 or Acinetobacter baumannii ATCC 19606.

9. A preparation containing the pyrazole cyanoacetylene pyridone acid compound and its pharmaceutically acceptable salt according to any one of claims 1-4.

10. The formulation of claim 9, wherein, The preparation is one of a tablet, a capsule, a granule, an injection, a powder injection, an eye drop, a liniment, a suppository, an ointment or an aerosol.

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