A thiazolamide MBL inhibitor and its preparation method and use

Abstract

The present invention provides a thiazolylamine MBL inhibitor and a preparation method and use thereof, belonging to the field of pharmaceutical chemistry. The thiazolylamine MBL inhibitor is a compound shown in formula I, or a salt thereof, or an isomer thereof, or a hydrate thereof, or a solvate thereof, or a prodrug thereof. The compound of the present invention has good inhibitory activity against metallo-β-lactamase (MBL), especially against multiple clinically relevant MBL subtypes, such as VIM, NDM, IMP and other subtypes, has strong Gram-negative bacterial permeability, can be used to prepare a broad-spectrum MBL inhibitor, and has good development prospects as a drug molecule to overcome antibacterial resistance.

Description

Technical Field

[0001] The present invention belongs to the field of pharmaceutical chemistry, and specifically relates to a thiazolamine MBL inhibitor and a preparation method and use thereof. Background Art

[0002] β-lactam antibiotics have the characteristics of broad antibacterial spectrum, high safety and good clinical efficacy, and are currently the most commonly used antibiotics. However, due to the abuse and irrational use of antibiotics, bacteria have developed serious resistance to antibiotics, and human health is seriously threatened. In 2019, the World Health Organization listed the problem of antimicrobial resistance as one of the top ten health threats in the world. Among the resistant pathogenic microorganisms, the problem of antibiotic resistance of Gram-negative bacteria is particularly severe. Gram-negative bacteria such as Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter that are resistant to carbapenem antibiotics have been listed as extremely dangerous pathogens by the World Health Organization. Current studies have shown that there are many mechanisms of resistance of Gram-negative bacteria, including preventing antibiotics from entering bacteria, expelling antibiotics through efflux pumps, changing the target of antibiotic action, and hydrolyzing antibiotics. Among them, the production of β-lactamase to hydrolyze antibiotics is its main mechanism against β-lactam antibiotics. Based on differences in amino acid sequences and catalytic mechanisms, β-lactamases can be divided into two categories: serine β-lactamase (SBL) and metallo-β-lactamase (MBL). Developing β-lactamase inhibitors and combining them with β-lactam antibiotics to inhibit the hydrolysis of β-lactamases and restore the antibacterial activity of antibiotics is an effective treatment strategy for combating resistant Gram-negative bacterial infections. To date, six SBL inhibitors have been successfully marketed, while there are no clinically available inhibitors for MBL.

[0003] Based on differences in amino acid sequence and active site structure, MBL can be further divided into three subclasses: B1, B2, and B3. Among them, the B1 class MBL includes the most clinically relevant VIM, NDM, and IMP subtypes, which are found in almost all major Gram-negative pathogens, including Klebsiella pneumoniae, Pseudomonas aeruginosa, Acinetobacter baumannii, and other common clinical pathogens. MBL has a wide range of substrate specificity and can hydrolyze all β-lactam antibiotics and β-lactam SBL inhibitors except monocyclic ones. Some MBL can even hydrolyze the non-β-lactam SBL inhibitor avibactam. Recently, it has been reported that NDM-1 can slowly hydrolyze the monocyclic β-lactam antibiotic aztreonam, which may mean that MBL has acquired the ability to hydrolyze monocyclic β-lactam antibiotics in the process of continuous evolution. In addition, some MBL genes exist on bacterial mobile gene elements and can be rapidly and widely spread among bacteria through horizontal gene transfer. Therefore, the production of MBL enzymes in bacteria is one of the most worrying factors of drug resistance, and the development of effective MBL inhibitors is urgent and of great clinical significance.

[0004] At present, many types of MBL inhibitors have been reported, including thiol, carboxylic acid, phosphate, etc. However, due to the large number of MBL subtypes, the large differences in structure, and the presence of flexible and changeable loops in the active site, most inhibitors lack broad-spectrum inhibitory activity against multiple MBL subtypes, especially clinically relevant VIM, NDM, IMP and other subtypes. In addition, the low permeability of the bacterial outer membrane is also a common disadvantage of the MBL inhibitors reported so far. Therefore, it is urgent to develop MBL inhibitors with broad-spectrum inhibitory activity and good antibacterial activity to provide new lead compounds for the research and development of clinically effective MBL inhibitors. Summary of the invention

[0005] The purpose of the present invention is to provide a thiazolamine MBL inhibitor and a preparation method and use thereof.

[0006] The present invention provides a compound represented by formula I, or a salt thereof, or an isomer thereof, or a hydrate thereof, or a solvate thereof, or a prodrug thereof:

[0007]

[0008] in,

[0009] R1 is selected from hydrogen, C1-C8 alkyl, halogen, hydroxy, carboxyl, amino, nitro, -NHR1';

[0010] R1' is selected from C1-C8 alkyl substituted by 0-5 R3, -C(O)R3';

[0011] R3 are independently selected from 5-10 membered aryl and 5-10 membered heteroaryl;

[0012] R3' is selected from C1-C8 alkyl;

[0013] L is selected from none, C1-C8 alkylene substituted with 0-5 R4, -S-, -O-,

[0014] R4 are independently selected from C1 to C8 alkyl;

[0015] R2 is selected from C1-C8 alkyl, C2-C8 alkenyl, C2-C8 alkynyl, 5-10 membered aryl substituted by 0-5 R5, 3-10 membered heteroaryl substituted by 0-5 R5, 3-10 membered heterocyclyl substituted by 0-5 R5, 3-10 membered cycloalkyl substituted by 0-5 R5;

[0016] R5 is independently selected from C1-C8 alkyl substituted by 0-5 R6, C1-C8 alkoxy substituted by 0-5 R6, halogen, carboxyl, nitro, amino, hydroxyl, -C(O)R7, -C(O)OR7, 5-10 membered aryl substituted by 0-5 R6, 5-10 membered heteroaryl substituted by 0-5 R6;

[0017] R6 is independently selected from C1-C8 alkyl, halogen, hydroxyl, carboxyl, amino, nitro, cyano, C2-C8 alkynyl, 5-10 membered aryl;

[0018] R7 is selected from C1-C8 alkyl, halogen, hydroxyl, carboxyl, amino, nitro, cyano, 5-10 membered aryl, 5-10 membered heteroaryl;

[0019] When R1 is selected from amino and R2 is selected from phenyl, L2 is not selected from C1 alkylene;

[0020] When R1 is selected from hydrogen and R2 is selected from phenyl, L2 is not selected from C1 alkylene, C1 alkylene substituted with one C1 alkyl, or C2 alkylene;

[0021] When R1 is selected from amino, L is selected from none, R2 is not selected from thienyl, C1-C5 alkyl;

[0022] When R1 is selected from amino, R2 is selected from C1 alkyl, L is not selected from C2-C4 alkylene, none, C2 alkylene substituted with 1 C1 alkyl, C1 alkylene substituted with 1 C1 alkyl;

[0023] When R1 is selected from hydrogen and R2 is selected from C1 alkyl, L is not selected from C2 alkylene substituted with 1 C1 alkyl, C1 alkylene substituted with 1 C1 alkyl;

[0024] When R1 is selected from amino, L is selected from C1 alkylene, R is not selected from

[0025] Further,

[0026] R1 is selected from hydrogen, C1-C8 alkyl, halogen, hydroxy, carboxyl, amino, nitro, -NHR1';

[0027] R1' is selected from C1-C8 alkyl substituted by 0-3 R3, -C(O)R3';

[0028] R3 are independently selected from phenyl, pyridyl, thienyl, pyrazolyl, furyl;

[0029] R3' is selected from C1-C8 alkyl;

[0030] L is selected from none, C1-C8 alkylene substituted with 0-3 R4, -S-, -O-,

[0031] R4 are independently selected from C1 to C8 alkyl;

[0032] R2 is selected from C1-C8 alkyl, C2-C8 alkenyl, C2-C8 alkynyl, phenyl substituted by 0-3 R5, thienyl substituted by 0-3 R5, piperidinyl substituted by 0-3 R5, azetidine substituted by 0-3 R5, naphthyl substituted by 0-3 R5;

[0033] R5 is independently selected from C1-C8 alkyl substituted by 0-3 R6, C1-C8 alkoxy substituted by 0-3 R6, halogen, carboxyl, nitro, amino, hydroxyl, -C(O)R7, -C(O)OR7, phenyl substituted by 0-3 R6, pyridyl substituted by 0-3 R6;

[0034] R6 is independently selected from C1-C8 alkyl, halogen, hydroxyl, carboxyl, amino, nitro, cyano, C2-C8 alkynyl, phenyl;

[0035] R7 is selected from C1-C8 alkyl, halogen, hydroxy, carboxyl, amino, nitro, cyano, phenyl, furyl.

[0036] Further,

[0037] R1 is selected from hydrogen, halogen, amino, -NHR1';

[0038] R1' is selected from C1-C3 alkyl substituted by 0-1 R3, -C(O)R3';

[0039] R3 are independently selected from phenyl, pyridyl, thienyl, pyrazolyl, furyl;

[0040] R3' is selected from C1-C3 alkyl;

[0041] L is selected from none, C1-C6 alkylene substituted with 0-1 R4, -S-, -O-,

[0042] R4 are independently selected from C1 to C3 alkyl;

[0043] R2 is selected from C1-C7 alkyl, C2-C4 alkenyl, C2-C4 alkynyl, phenyl substituted by 0-2 R5, thienyl substituted by 0-1 R5, piperidinyl substituted by 0-1 R5, azetidine substituted by 0-1 R5, naphthyl substituted by 0-1 R5;

[0044] R5 is independently selected from C1-C4 alkyl substituted by 0-3 R6, C1-C3 alkoxy substituted by 0-3 R6, halogen, carboxyl, nitro, amino, hydroxyl, -C(O)R7, -C(O)OR7, phenyl substituted by 0-1 R6, pyridyl substituted by 0-1 R6;

[0045] R6 is independently selected from C1-C3 alkyl, halogen, hydroxyl, carboxyl, amino, nitro, cyano, C2-C4 alkynyl, and phenyl;

[0046] R7 is selected from C1-C4 alkyl, halogen, hydroxy, carboxyl, amino, nitro, cyano, phenyl, furyl.

[0047] Furthermore, the compound is as shown in Formula II:

[0048]

[0049] in,

[0050] L is selected from none, C1-C6 alkylene substituted with 0-1 R4, -S-, -O-,

[0051] R4 are independently selected from C1 to C3 alkyl;

[0052] R2 is selected from C1-C7 alkyl, C2-C4 alkenyl, C2-C4 alkynyl, phenyl substituted by 0-2 R5, thienyl substituted by 0-1 R5, piperidinyl substituted by 0-1 R5, azetidine substituted by 0-1 R5, naphthyl substituted by 0-1 R5;

[0053] R5 is independently selected from C1-C4 alkyl substituted by 0-3 R6, C1-C3 alkoxy substituted by 0-3 R6, halogen, carboxyl, nitro, amino, hydroxyl, -C(O)R7, -C(O)OR7, phenyl substituted by 0-1 R6, pyridyl substituted by 0-1 R6;

[0054] R6 is independently selected from C1-C3 alkyl, halogen, hydroxyl, carboxyl, amino, nitro, cyano, C2-C4 alkynyl, and phenyl;

[0055] R7 is selected from C1-C4 alkyl, halogen, hydroxy, carboxyl, amino, nitro, cyano, phenyl, furyl.

[0056] Furthermore, the compound is as shown in Formula III:

[0057]

[0058] in,

[0059] R1 is selected from hydrogen, halogen, amino, -NHR1';

[0060] R1' is selected from C1-C3 alkyl substituted by 0-1 R3, -C(O)R3';

[0061] R3 are independently selected from phenyl, pyridyl, thienyl, pyrazolyl, furyl;

[0062] R3' is selected from C1-C3 alkyl;

[0063] L is selected from none, C1-C6 alkylene substituted with 0-1 R4, -S-, -O-,

[0064] R4 are independently selected from C1 to C3 alkyl;

[0065] R 5a , R 5b , R 5c Each of the following groups is independently selected from hydrogen, C1-C4 alkyl substituted by 0-3 R6, C1-C3 alkoxy substituted by 0-3 R6, halogen, carboxyl, nitro, amino, hydroxyl, -C(O)R7, -C(O)OR7, phenyl substituted by 0-1 R6, and pyridyl substituted by 0-1 R6;

[0066] R6 is independently selected from C1-C3 alkyl, halogen, hydroxyl, carboxyl, amino, nitro, cyano, C2-C4 alkynyl, and phenyl;

[0067] R7 is selected from C1-C4 alkyl, halogen, hydroxy, carboxyl, amino, nitro, cyano, phenyl, furyl.

[0068] Furthermore, the compound is as shown in Formula IV:

[0069]

[0070] in,

[0071] R1 is selected from hydrogen, halogen, amino, -NHR1';

[0072] R1' is selected from C1-C3 alkyl substituted by 0-1 R3, -C(O)R3';

[0073] R3 are independently selected from phenyl, pyridyl, thienyl, pyrazolyl, furyl;

[0074] R3' is selected from C1-C3 alkyl;

[0075] L is selected from none, C1-C6 alkylene substituted with 0-1 R4, -S-, -O-,

[0076] R4 are each independently selected from C1 to C3 alkyl.

[0077] Furthermore, the compound is one of the following compounds:

[0078]

[0079]

[0080] The present invention also provides the use of the aforementioned compound, or its salt, or its isomer, or its hydrate, or its solvate, or its prodrug in the preparation of MBL inhibitors and / or antibacterial drugs.

[0081] The present invention also provides a drug, which is a preparation prepared by taking the aforementioned compound, or its salt, or its isomer, or its hydrate, or its solvate, or its prodrug as an active ingredient and adding pharmaceutically acceptable excipients or auxiliary ingredients.

[0082] The present invention also provides a use of a thiazolamine compound, or a salt thereof, or an isomer thereof, or a hydrate thereof, or a solvate thereof, or a prodrug thereof in the preparation of an MBL inhibitor and / or an antibacterial drug, characterized in that the thiazolamine compound is one of the following compounds:

[0083]

[0084] The compounds and derivatives provided in the present invention can be named according to the IUPAC (International Union of Pure and Applied Chemistry) or CAS (Chemical Abstracts Service, Columbus, OH) nomenclature system.

[0085] Definitions of terms used in the present invention: Unless otherwise stated, the initial definitions provided for groups or terms in this document apply to the groups or terms throughout the specification; for terms that are not specifically defined in this document, the meaning that a person skilled in the art can give them should be given based on the disclosure and context.

[0086] "Substitution" refers to the replacement of a hydrogen atom in a molecule by another different atom or molecule.

[0087] The minimum and maximum carbon atom content in a hydrocarbon group is indicated by a prefix, for example, the prefix C a ~C b Alkyl refers to any alkyl group containing from "a" to "b" carbon atoms. Thus, for example, "C1-C8 alkyl" refers to an alkyl group containing 1 to 8 carbon atoms; "C1-C8 alkoxy" refers to an alkoxy group containing 1 to 8 carbon atoms.

[0088] "Alkyl" refers to a saturated hydrocarbon chain having a specified number of carbon atoms. For example, C1-C8 alkyl refers to an alkyl group having 1 to 8 carbon atoms, i.e., 1, 2, 3, 4, 5, 6, 7 or 8 carbon atoms. Alkyl groups can be straight or branched. Representative branched alkyl groups have one, two or three branches. Alkyl groups include methyl, ethyl, propyl (n-propyl and isopropyl), butyl (n-butyl, isobutyl and tert-butyl), pentyl (n-pentyl, isopentyl and neopentyl) and hexyl, etc.

[0089] "Halogen" is fluorine, chlorine, bromine or iodine.

[0090] The compounds of the present invention have good inhibitory activity against metallo-β-lactamase (MBL), especially against multiple clinically relevant MBL subtypes, such as VIM, NDM, IMP and other subtypes, have strong Gram-negative bacterial permeability, can be used to prepare broad-spectrum MBL inhibitors, and have good development prospects as drug molecules for overcoming antimicrobial resistance.

[0091] Obviously, according to the above contents of the present invention, in accordance with common technical knowledge and customary means in the art, without departing from the above basic technical ideas of the present invention, other various forms of modification, replacement or change may be made.

[0092] The above contents of the present invention are further described in detail below through specific implementation methods in the form of embodiments. However, this should not be understood as the scope of the above subject matter of the present invention being limited to the following examples. All technologies realized based on the above contents of the present invention belong to the scope of the present invention. DETAILED DESCRIPTION

[0093] Unless otherwise specified, the raw materials and equipment used in the specific embodiments of the present invention are all known products and are obtained by purchasing commercially available products.

[0094] Example 1. Synthesis of Compounds 1, 2, 11-14, 18, 20-22, 26, 28, 29, 39, 40, 42, 45, 48, 49, 67-69 and 71-74

[0095] Synthesis route: The substrate is the corresponding aldehyde

[0096]

[0097] 1. Synthesis of compound 39

[0098]

[0099] Ethyl dichloroformate (1 eq) and n-hexanal (0.8 eq) were dissolved in ether (10 ml), and sodium methoxide (1.1 eq) was added dropwise at 0°C. After the addition was completed, the mixture was stirred for two hours and then stirred at room temperature for 5 hours. Saturated brine (about 50 ml) was added to the reaction solution, and then extracted with ether (3×30 ml). After concentration, the mixture was dissolved in methanol, and thiourea (0.8 eq) was added. The mixture was refluxed at 65°C for 5 hours to stop the reaction. The methanol was dried by spin drying, dissolved in water, adjusted to slightly alkaline (pH=8) with aqueous ammonia, extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to silica gel column chromatography (PE:EA=1:1, v / v) to obtain intermediate a (yellow solid, yield about 70%).

[0100] Dissolve intermediate a (1 eq) in ethanol and water (volume ratio = 3:1) and add sodium hydroxide (3 eq). After reacting at room temperature for 2 hours, stop the reaction. Adjust the reaction solution to slightly acidic (pH = 5), spin dry, and perform silica gel column chromatography (DCM: CH3OH = 30:1, v / v). Compound 39 (yield 60%) was obtained.

[0101] Compound 39: 1 H NMR (400MHz, DMSO-d6) δ6.98 (s br, 2H), 3.03-2.98 (m, 2H), 1.54-1.50 (m, 2H), 1.38-1.24 (m, 4H), 0.86 (t, J = 6.4Hz, 3H) ppm. 13 C NMR (101MHz, DMSO-d6) δ164.37,163.99,137.86,31.27,31.13,26.62,22.25,14.32ppm.HRMS:m / z calcd forC9H 14 N2O2S[M+H] + 215.0849, found 215.0843; [M-H2O+H] + 197.0749, found 197.0737.

[0102] 2. Synthesis of compounds 1, 2, 11-14, 18, 20-22, 26, 28, 29, 40, 42, 45, 48, 49, 67-69 and 71-74

[0103] According to the synthesis method described in compound 39, the raw material n-hexanal was replaced by phenylacetaldehyde to prepare compound 1, the raw material n-hexanal was replaced by 2-phenylpropanal to prepare compound 2, the raw material n-hexanal was replaced by n-valeraldehyde to prepare compound 11, the raw material n-hexanal was replaced by n-heptylaldehyde to prepare compound 12, the raw material n-hexanal was replaced by n-octanal to prepare compound 13, the raw material n-hexanal was replaced by n-butyraldehyde to prepare compound 14, the raw material n-hexanal was replaced by p-nitrophenylacetaldehyde to prepare compound 18, the raw material n-hexanal was replaced by acetaldehyde to prepare compound 20, the raw material n-hexanal was replaced by thiophene-2-acetaldehyde to prepare compound 21, the raw material n-hexanal was replaced by isobutylaldehyde to prepare compound 22, the raw material n-hexanal was replaced by 2-methylbutyraldehyde to prepare compound 26, the raw material n-hexanal was replaced by formaldehyde to prepare compound 28, the raw material n-hexanal was replaced by 3-methylbutyraldehyde to prepare compound 29, the raw material n-hexanal was replaced by 4-(2-oxoethyl)piperidine-1-carboxylic acid tert-butyl Compound 40 was prepared by replacing the raw material n-hexanal with 4-pentenal to prepare compound 42, the raw material n-hexanal was replaced with 2-(1-benzoylpiperidin-4-yl)acetaldehyde to prepare compound 45, the raw material n-hexanal was replaced with 4-aminophenylacetaldehyde to prepare compound 48, the raw material n-hexanal was replaced with 2-(1-furanoylpiperidin-4-yl)acetaldehyde to prepare compound 49, the raw material n-hexanal was replaced with 3-(4-isopropylphenyl)-2-methylpropanal to prepare compound 67 , the raw material n-hexanal was replaced by 3-(3-trifluoromethylphenyl) propionaldehyde to prepare compound 68, the raw material n-hexanal was replaced by lily of the valley aldehyde to prepare compound 69, the raw material n-hexanal was replaced by 3,5-dichlorophenylacetaldehyde to prepare compound 71, the raw material n-hexanal was replaced by 3,4-dichlorophenylacetaldehyde to prepare compound 72, the raw material n-hexanal was replaced by 3,5-dibromophenylacetaldehyde to prepare compound 73, and the raw material n-hexanal was replaced by 3-chloro-5-bromo-phenylacetaldehyde to prepare compound 74. The structures of each compound are shown below:

[0104]

[0105]

[0106] The characterization data of each compound are shown below:

[0107] Compound 1: 1 H NMR(400MHz,DMSO-d6)δ7.33-7.20(m,7H),4.34(s,2H)ppm. 13C NMR(101MHz,DMSO-d6)δ165.26,163.57,140.51,136.16,136.04,129.00,128.78,127.00,32.50ppm.HRMS:m / z calcd for C 11 H 10 N2O2S[M+H] + 235.0536, found 235.0541; [M-H2O+H] + 217.0436, found 217.0433.

[0108] Compound 2: 1 H NMR (400MHz, DMSO-d6) δ7.33-7.28(m,4H),7.24-7.18(m,1H),7.08(sbr,2H),5.22(q,J=7.2Hz,1H),1.54(d,J=7.2Hz,3H)ppm. 13 C NMR(101MHz,DMSO-d6)δ164.96,163.94,145.50,143.11,136.08,128.91,127.28,126.94,36.74,23.08ppm.HRMS:m / z calcd for C 12 H 12 N2O2S[M+H] + 249.0692, found 249.0684; [M-H2O+H] + 231.0592, found 231.0578.

[0109] Compound 11: 1 H NMR(400MHz,DMSO-d6)δ12.23(s br,1H),6.88(s,2H),2.97(t,J=2.4Hz,2H),1.54-1.47(m,2H),1.34-1.27(m,2H),0.88(t,J=6.8Hz,3H)ppm.HRMS:m / zcalcd for C8H 12 N2O2S[M+H] + 201.0692, found 201.0694; [M-H2O+H] + 183.0592,found183.0573;[M+Na] + 223.0517, found 223.0508.

[0110] Compound 12:1 H NMR(400MHz, DMSO-d6)δ6.89(s,2H),2.96(t,J=8.0Hz,2H),1.51(t,J=8.0Hz,2H),1.26-1.23(m,6H),0.86-0.82(m,3H)ppm.HRMS:m / z calcd for C 10 H 16 N2O2S[M+H] + 229.1005, found 229.0995; [M-H2O+H] + 211.0905, found 211.0887; C 10 H 16 N2O2S[M+Na] + 251.0830, found 251.0521.

[0111] Compound 13: 1 H NMR (400MHz, DMSO-d6) δ6.88(s,2H),2.96(t,J=8.0Hz,2H),1.51(s,2H),1.25(d,J=7.2Hz,8H),0.85(d,J=6.0Hz,3H)ppm.HRMS:m / z calcd for C 11 H 18 N2O2S[M+H] + 243.1162, found 243.1147; [M-H2O+H] + 225.1062, found 225.1042.

[0112] Compound 14: 1 H NMR(400MHz, DMSO-d6)δ8.68(s,2H),2.97(t,J=7.2Hz,2H),1.61-1.54(m,2H),0.91(t,J=7.2Hz,3H)ppm.HRMS:m / z calcd for C7H 10 N2O2S[M+H] + 187.0536, found 187.0536; [M-H2O+H] + 169.0436, found 169.0421.

[0113] Compound 18: 1 H NMR (400MHz, DMSO-d6) δ8.14 (d, J = 8.4Hz, 2H) 7.76-7.53 (m, br, 4H) ppm.

[0114] Compound 20: 1 H NMR (400MHz, DMSO-d6) δ6.71 (s, 2H), 2.31 (s, 3H) ppm.

[0115] Compound 21: 1 H NMR(400MHz,DMSO-d6)δ8.82(s br,2H),7.43(d,J=4.8Hz,1H),7.02-6.98(m,2H),4.59(s,2H)ppm.HRMS:m / z calcd for C9H8N2O2S2[M+H] + 241.0100,found241.0097;[M-H2O+H] + 223.0000, found 222.9991.

[0116] Compound 22: 1 H NMR(400MHz,DMSO-d6)δ7.88-7.60(m,1H),6.97(s,1H),4.25(s,1H),1.14(s,6H)ppm.HRMS:m / z calcd for C7H 10 N2O2S[M+H] + 187.0536, found 187.0559; [M-H2O+H] + 169.0436, found 169.0449.

[0117] Compound 26: HRMS: m / z calcd for C8H 12 N2O2S[M+H] + 201.0692, found 201.0702; [M-H2O+H] + 183.0592, found 183.0592; [M+Na] + 223.0517, found 223.0527.

[0118] Compound 28: 1 H NMR(400MHz,DMSO-d6)δ7.20(s,2H),7.00(s,1H)ppm.HRMS:m / zcalcd for C4H4N2O2S[M+H] + 145.0066, found 145.0067; [M-H2O+H] + 126.9966,found126.9958;[M+Na]+ 166.9881, found 166.9891.

[0119] Compound 29: 1 H NMR (400MHz, DMSO-d6) δ6.97 (s, 2H), 2.94 (d, J = 6.8Hz, 2H), 1.78-1.71 (m, 1H), 0.88 (d, J = 6.8Hz, 6H) ppm.HRMS: m / z calcd for C8H 12 N2O2S[M+H] + 201.0692, found 201.0688; [M-H2O+H] + 183.0592, found 183.0579; [M+Na] + 223.0517, found 223.0514.

[0120] Compound 40: 1 H NMR(400MHz,DMSO-d6)δ3.88(s br,2H),3.03-3.00(m,2H),2.67(sbr,2H),1.61-1.56(m,3H),1.38(m,9H),1.05-1.02(m,2H)ppm.HRMS:m / z calcd forC 15 H 23 N3O4S[M+H] + 342.1482,found 342.1486.[M+Na] + 364.1307, found 364.1298.

[0121] Compound 42: 1 H NMR (400MHz, DMSO-d6) δ8.78(s br,2H),5.86-5.76(m,1H),5.08-5.00(m,2H),3.10(t,J=7.2Hz,2H),2.31(t,J=7.2Hz,2H)ppm. 13 C NMR(101MHz,DMSO-d6)δ166.76,160.37,137.10,135.71,116.85,34.45,25.84ppm.HRMS:m / z calcd for C8H 10 N2O2S[M+H] + 199.0536, found 199.0533; [M-H2O+H] + 181.0436, found 181.0428.

[0122] Compound 45: HRMS: m / z calcd for C 17 H 19 N3O3S[M+H] + 346.1220, found 346.1211.

[0123] Compound 48: LC-MS m / z: 250.1 [M+H] + .

[0124] Compound 49: LC-MS m / z: 236.1 [M+H] + .

[0125] Compound 67: 1 H NMR(400MHz,DMSO-d6)δ7.13-7.08(m,4H),6.88(s,2H),4.14(s,1H),2.86-2.76(m ,2H),2.68-2.63(m,1H),1.17(d,J=6.8Hz,6H),1.12(d,J=6.8Hz,3H)ppm.HRMS:m / z calcd for C 16 H 20 N2O2S[M+H] + 305.1318, found 305.1312.

[0126] Compound 68: 1 H NMR (400MHz, DMSO-d6) δ7.58-7.51(m,4H),6.89(s,2H),3.31(t,J=8.0Hz,2H),2.95(t,J=8.0Hz,2H)ppm.HRMS:m / z calcd for C 13 H 11 F3N2O2S[M+H] + 317.0566, found 317.0528.

[0127] Compound 69: 1 H NMR(400MHz,DMSO-d6)δ7.86(s br,2H),7.29(d,J=7.6Hz,2H),7.10(d,J=7.6Hz,2H),4.18-4.11(m,1H),2.83-2 .78(m,1H),2.73-2.68(m,1H),1.25(s,9H),1.15(d,J=6.8Hz,3H)ppm.HRMS:m / z calcd for C 17 H22 N2O2S[M+H] + 319.1475, found 319.1461.

[0128] Compound 71: 1 H NMR (400MHz, DMSO-d6) δ8.60 (s br, 2H), 7.50 (s, 1H), 7.38 (d, J = 1.6Hz, 2H), 4.39 (s, 2H) ppm. HRMS: m / z calcd for C 11 H8Cl2N2O2S[M+H] + 302.9756,found302.9757,304.9731.

[0129] Compound 72: HRMS: m / z calcd for C 11 H8Cl2N2O2S[M+H] + 302.9756,found 302.9752,304.9729.

[0130] Compound 73: HRMS: m / z calcd for C 11 H8Br2N2O2S[M+H] + 390.8746,found 390.8749,392.8722,394.8707.

[0131] Compound 74: HRMS: m / z calcd for C 11 H8BrClN2O2S[M+H] + 346.9251,found346.9250,348.9221.

[0132] Example 2. Synthesis of Compounds 5, 17, 43, 46, 47, 50, 52, 55, 58, 60, 64, 65, 66

[0133] Synthesis route: The substrate is the corresponding carboxylic acid

[0134]

[0135] 1. Synthesis of compound 43

[0136]

[0137] The raw material 1-(tert-butoxycarbonyl)azetidine-3-acetic acid (1 eq) was placed in a reaction bottle, dissolved with dry tetrahydrofuran, and then lithium aluminum hydroxide (1.5 eq) was slowly added under an ice bath. After the addition was completed, the reaction was moved to room temperature and monitored by TLC. After the reaction was completed, the reaction was stopped, and 1% of the volume of water (such as adding 200 mg of lithium aluminum hydroxide and adding 2 ml of water) was added under an ice bath. Then, a sodium hydroxide aqueous solution with a mass fraction of 10% was added with the same volume of water, and then twice the volume of water was added before stirring for about 15 minutes, and then a small amount of anhydrous magnesium sulfate was added. After stirring for 15 minutes, the mixture was filtered and the tetrahydrofuran was dried by spin drying. The remaining solution was extracted 3 times with a small amount of water and ethyl acetate (the volume ratio of water to ethyl acetate was 1:2). The organic phase was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and spin dried to obtain intermediate a (yield of about 50-70%).

[0138] Place intermediate a (1 eq) in a reaction bottle, dissolve it with dichloromethane, add dessmartin oxidant (1.2 eq), stir at room temperature for about 1 h, monitor by TLC, and stop the reaction when the reaction is complete. Filter the filtrate with a cylindrical funnel, dry and concentrate the filtrate under reduced pressure, add fine silica gel, spin dry and pass through a column, and PE to obtain intermediate b (yield of about 40-60%).

[0139] After dissolving ethyl dichloroformate (1 eq) and intermediate b (0.8 eq) in ether, sodium methoxide (1.1 eq) was added dropwise at 0°C. After stirring for two hours, the mixture was moved to room temperature and stirred for 5 hours. An appropriate amount of saturated brine (50 ml) was added, and then extracted with ether (3×30 ml). The organic phase was concentrated and dissolved in methanol. Thiourea (0.8 eq) was added and refluxed at 65°C for 5 hours to stop the reaction. The methanol was dried by spin drying, dissolved in water, adjusted to slightly alkaline (pH=8) with aqueous ammonia, extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and chromatographed on a silica gel column (PE:EA=1:1, v / v) to obtain intermediate c (yellow solid, yield of about 50-80%).

[0140] Dissolve intermediate c (1 eq) in ethanol and water (volume ratio 3:1) and add sodium hydroxide (3 eq). After reacting at room temperature for 2 hours, stop the reaction. Adjust the reaction solution to slightly acidic (pH = 5), spin dry, and perform silica gel column chromatography (DCM: CH3OH = 30:1, v / v). Compound 43 (yield 60%) was obtained. LC-MS m / z: 314.1 [M+H] + .

[0141] 2. Synthesis of compounds 5, 17, 46, 47, 50, 52, 55, 58, 60, 64, 65, and 66

[0142] According to the synthesis method described in compound 43, the raw material 1-(tert-butoxycarbonyl)azetidine-3-acetic acid was replaced by phenylpropionic acid to prepare compound 5, the raw material 1-(tert-butoxycarbonyl)azetidine-3-acetic acid was replaced by p-nitrophenylacetic acid to prepare compound 17, the raw material 1-(tert-butoxycarbonyl)azetidine-3-acetic acid was replaced by 3-nitrophenylacetic acid to prepare compound 46, the raw material 1-(tert-butoxycarbonyl)azetidine-3-acetic acid was replaced by 3-aminophenylacetic acid to prepare compound 47, the raw material 1-(tert-butoxycarbonyl)azetidine-3-acetic acid was replaced by 4-biphenylacetic acid to prepare compound 50, the raw material 1-(tert-butoxycarbonyl)azetidine-3-acetic acid was replaced by 4-(trifluoromethoxy)phenylacetic acid to prepare To compound 52, the raw material 1-(tert-butoxycarbonyl)azetidine-3-acetic acid is replaced by 4-trifluoromethylphenylacetic acid to prepare compound 55, the raw material 1-(tert-butoxycarbonyl)azetidine-3-acetic acid is replaced by 4-isopropylphenylacetic acid to prepare compound 58, the raw material 1-(tert-butoxycarbonyl)azetidine-3-acetic acid is replaced by 4-propylphenylacetic acid to prepare compound 60, the raw material 1-(tert-butoxycarbonyl)azetidine-3-acetic acid is replaced by 3-(2-thiophene)propionic acid to prepare compound 64, the raw material 1-(tert-butoxycarbonyl)azetidine-3-acetic acid is replaced by naproxen to prepare compound 65, and the raw material 1-(tert-butoxycarbonyl)azetidine-3-acetic acid is replaced by ibuprofen to prepare compound 66. The structures of each compound are shown below:

[0143]

[0144] The characterization data of each compound are shown below:

[0145] Compound 5: 1 H NMR (400MHz, DMSO-d6) δ7.28-7.21(m,7H),3.27(t,J=8.0Hz,2H),2.84(t,J=8.0Hz,2H)ppm. 13 C NMR(101MHz,DMSO-d6)δ164.71,163.30,141.09,136.45,135.74,128.85,128.79,126.60,37.17,28.66ppm.HRMS:m / z calcd for C 12 H 12 N2O2S[M+H] + 249.0692, found 249.0689; [M-H2O+H] + 231.0592, found 231.0579; [M+Na]+ 271.0512, found 271.0512.

[0146] Compound 17: 1 H NMR (400MHz, DMSO-d6) δ8.21(d,J=8.8Hz,2H),7.68(d,J=9.2Hz,2H),7.56(s,2H),3.67(s,2H)ppm.HRMS:m / z calcd fo C 11 H9N3O4S[M+H] + 280.0387, found 280.0380.

[0147] Compound 46: 1 H NMR (400MHz, DMSO-d6) δ8.14(s,1H),8.08(d,J=6.8Hz,1H),7.74(d,J=6.8Hz,1H),7.61(t,J=7.2Hz,1H),7.06(s,2H),4.58(s,2H)ppm.HRMS:m / z calcd forC 11 H9N3O4S[M+H] + 280.0387, found 280.0382.

[0148] Compound 47: 1 H NMR (400MHz, DMSO-d6) δ6.94-6.90(m,2H),6.41-6.36(m,3H),6.19(s,2H)ppm. 13 C NMR (101MHz, DMSO-d6) δ164.79,149.21,141.21,137.34,129.36,116.25,114.16,112.62, 32.90ppm.HRMS:m / z calcd for C 11 H 11 N3O2S[M+H] + 250.0645,found250.0644; [M-H2O+H] + 232.0545, found 232.0539.

[0149] Compound 50: 1 H NMR (400MHz, DMSO-d6) δ7.65-7.59(m,4H),7.45(t,J=7.6Hz,2H),7.37-7.32(m,3H),7.04(s br,2H)ppm. 13C NMR (101MHz, DMSO-d6) δ165.13,163.95,140.37,139.98,138.85,137.13,136.01,129.38,129.33,127.78,127.28,127.02,32.12ppm.HRMS:m / z calcd for C 17 H 14 N2O2S[M+H] + 311.0849, found 311.0843.

[0150] Compound 52: 1 H NMR (400MHz, DMSO-d6) δ7.37 (d, J = 8.8 Hz, 2H), 7.29 (d, J = 8.8 Hz, 2H), 7.02 (s br, 2H), 4.38 (s, 2H) ppm. 13 C NMR(101MHz,DMSO-d6)δ165.31,164.05,147.37,140.25,130.51,121.82,121.54,119.28,116.73,31.66ppm.HRMS:m / z calcd forC 12 H9N3O2F3S[M+H] + 319.0359, found 319.0373.

[0151] Compound 55: 1 H NMR (400MHz, DMSO-d6) δ7.68 (d, J = 8.0 Hz, 2H), 7.56 (s br, 2H), 7.48 (d, J = 8.0 Hz, 2H), 4.46 (s, 2H) ppm. 13 C NMR (101MHz, DMSO-d6) δ165.72,163.01,145.07,134.25,129.57,127.90,127.58,125.88,123.43,32.03ppm.

[0152] Compound 58: 1 H NMR (400MHz, DMSO-d6) δ7.17-7.14(m,4H),6.90(s,2H),4.29(s,2H),2.87-2.80(m,1H),1.17(d,J=6.8Hz,6H)ppm. 13C NMR (101MHz, DMSO-d6) δ164.88,164.18,146.94,138.24,129.39,128.67,126.82,118.77,33.52,32.14,24.39ppm.HRMS:m / z calcdfor C 14 H 16 N2O2S[M+H] + 277.1005, found 277.0992.

[0153] Compound 60: 1 H NMR (400MHz, DMSO-d6) δ8.92(s br,1H),6.91(s,2H),4.29(s,2H),2.51(t,J=7.6Hz,2H),1.60-1.51(m,2H),0.87(t,J=7.6Hz,3H)ppm. 13 C NMR (101MHz, DMSO-d6) δ164.93,164.11,140.66,138.06,137.29,136.69,128.88,128.61,37.35,32.21,24.58,14.13ppm.HRMS:m / z calcd for C 14 H 16 N2O2S[M+H] + 277.1005, found 277.0998.

[0154] Compound 64: 1 H NMR (400MHz, DMSO-d6) δ7.32(d,J=4.8Hz,1H),6.94-6.92(m,2H),6.87(d,J=6.4Hz,2H),3.39-3.37(m,2H),3.05(t,J=7.6Hz,2H)ppm.HRMS:m / z calcd forC 10 H 10 N2O2S2[M+H] + 255.0256, found 255.0254; [M-H2O+H] + 237.0156, found 237.0148.

[0155] Compound 65: 1H NMR(400MHz,DMSO-d6)δ8.24(s br,2H),7.83(d,J=9.2Hz,1H),7.79-7.73(m,2H),7.41(d,J=8.4Hz,1H),7.29(s,1H),7.16(dd, J=9.2Hz,J=2.0Hz,1H),5.32(t,J=7.6Hz,1H),3.86(s,3H),1.67(d,J=7.2Hz,3H)ppm.HRMS:m / z calcd forC 17 H 16 N2O3S[M+H] + 329.0954, found 329.0947.

[0156] Compound 66: 1 H NMR(400MHz,DMSO-d6)δ7.19(d,J=8.0Hz,2H),7.08(d,J=8.0Hz,2H),6.92(s,2H),5.19-5.17(m,1H) ,2.40(d,J=7.2Hz,2H),1.82-1.74(m,1H),1.52(d,J=6.8Hz,3H),0.84(d,J=6.8Hz,6H)ppm.HRMS:m / z calcd for C 16 H 20 N2O2S[M+H] + 305.1318, found 305.1299.

[0157] Example 3. Synthesis of Compounds 51, 54, 57, and 62

[0158] Synthesis route: The substrate is the corresponding alcohol

[0159]

[0160] 1. Synthesis of Compound 51

[0161]

[0162] 3-Methoxyphenylethanol (1 eq) was placed in a reaction bottle, dissolved with dichloromethane (20 ml), and dessmartin oxidant (1.2 eq) was added. The reaction was stirred at room temperature for about 1 h and monitored by TLC. After the reaction was completed, the reaction was stopped and filtered through a cylindrical funnel. The filtrate was dried and concentrated under reduced pressure. Fine silica gel was added, and the mixture was spin-dried and passed through a column. PE was used to obtain intermediate a (yield of about 60%).

[0163] Ethyl dichloroformate (1 eq) and intermediate a (0.8 eq) were dissolved in ether (10 ml), and sodium methoxide (1.1 eq) was added dropwise at 0°C. After the addition was completed, the mixture was stirred for two hours and then stirred at room temperature for 5 hours. Saturated brine (50 ml) was added to the reaction solution, which was then extracted with ether (3×30 ml). The organic phase was concentrated and dissolved in methanol. Thiourea (0.8 eq) was added and refluxed at 65°C for 5 hours to stop the reaction. The methanol was dried by spin drying, dissolved in water, adjusted to slightly alkaline (pH=8) with aqueous ammonia, extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and chromatographed on a silica gel column (PE:EA=1:1, v / v) to obtain intermediate b (yellow solid, yield of about 50%).

[0164] The intermediate b (1 eq) was dissolved in ethanol and water (volume ratio = 3:1), and then sodium hydroxide (3 eq) was added. After reacting at room temperature for 2 hours, the reaction was stopped. The reaction solution was adjusted to slightly acidic (pH = 5), dried by spin drying, and chromatographed on a silica gel column (DCM: CH3OH = 30:1, v / v). Compound 51 (yield 60%) was obtained.

[0165] Compound 51: 1 H NMR (400MHz, DMSO-d6) δ8.20 (s br, 2H), 7.28 (t, J = 8.0Hz, 1H), 6.89-6.85 (m, 3H), 4.37 (s, 2H), 3.78 (s, 3H) ppm. 13 C NMR (101MHz, DMSO-d6) δ166.13,162.01,159.90,141.19,135.63,131.98,130.19,121.09,114.77,112.54,55.48,32.34ppm.HRMS:m / z calcd for C 12 H 12 N2O3S[M+H] + 265.0641, found 265.0637.

[0166] 2. Synthesis of compounds 54, 57, and 62

[0167] According to the synthesis method described for compound 51, the raw material 3-methoxyphenethanol was replaced by thiophene-3-ethanol to prepare compound 54, the raw material 3-methoxyphenethanol was replaced by 3-bromophenethanol to prepare compound 57, and the raw material 3-methoxyphenethanol was replaced by p-fluorophenethanol to prepare compound 62. The structures of each compound are shown below:

[0168]

[0169] The characterization data of each compound are shown below:

[0170] Compound 54: 1 H NMR (400MHz, DMSO-d6) δ7.46-7.45 (m, 1H), 7.22 (s, 1H), 7.03 (s, 2H), 6.99 (d, J = 4.4Hz, 1H), 4.34 (s, 2H) ppm. 13 C NMR(101MHz,DMSO-d6)δ164.93,164.19,140.94,137.41,135.79,128.69,126.71,122.16,27.50ppm.HRMS:m / z calcd forC9H8N2O2S2[M+H] + 241.0100, found 241.0101.

[0171] Compound 57: 1 H NMR(400MHz, DMSO-d6)δ7.45-7.41(m,2H),7.26(d,J=4.0Hz,2H),7.01-6.90(m,2H),4.44-4.36(m,2H)ppm.HRMS:m / z calcd for C 11 H9BrN2O2S[M+H] + 312.9641,found 312.9652,314.9628.

[0172] Compound 62: 1 H NMR(400MHz,DMSO-d6)δ7.76(s br,2H),7.32-7.29(m,2H),7.17-7.11(m,2H),4.34(s,2H)ppm.HRMS:m / z calcd for C 11 H9FN2O2S[M+H] + 253.0442,found253.0434;[M-H2O+H] + 235.0341, found 235.0327.

[0173] Example 4. Synthesis of Compounds 41 and 44

[0174] Compound 44 and compound 41 were obtained by removing Boc from compound 43 and compound 40, respectively.

[0175] 1. Synthesis of compound 44

[0176]

[0177] Compound 43 was dissolved in dichloromethane (10 ml) in a reaction bottle, trifluoroacetic acid (1 ml) was added under ice bath, and after addition, the mixture was stirred at room temperature for several hours, and the reaction was completed by TLC monitoring. The reaction was stopped, the solvent was dried, water (30 ml) was added to dissolve, and the mixture was adjusted to weak alkalinity (pH=8) with saturated sodium bicarbonate, extracted with ethyl acetate (3×30 ml), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (DCM:CH3OH=50:1; 30:1; 10:1, v / v) to obtain compound 44 with a yield of about 80%. LC-MS m / z: 214.1 [M+H] + .

[0178] 2. Synthesis of Compound 41

[0179] According to the synthesis method of compound 44, the raw material compound 43 was replaced by compound 40 to prepare compound 41. The structure of compound 41 is shown below:

[0180]

[0181] The characterization data of compound 41 are as follows: HRMS: m / z calcd for C 10 H 15 N3O2S[M+H] + 242.0958, found 242.0956.

[0182] Example 5. Synthesis of Compounds 53, 56, 61 and 63

[0183] Synthesis route:

[0184]

[0185] Wherein, X is a bromine atom, and R is a different substituent.

[0186] 1. Synthesis of compound 53

[0187]

[0188] Place intermediate a (1 eq) in a reaction flask, dissolve it with DCM (20 ml), and slowly drop boron tribromide (3 eq) in an ice bath. After the addition is completed, stir at room temperature for a few hours, monitor by TLC, and the reaction is completed. Stop the reaction, spin dry the solvent, add a small amount of water (30 ml), and adjust to weak alkalinity (pH = 8) with saturated sodium bicarbonate, extract with ethyl acetate (3×30 ml), dry over anhydrous sodium sulfate, concentrate under reduced pressure, and perform column chromatography with PE:EA=(1:1, v / v) to obtain intermediate b as a yellow solid (yield of about 30%).

[0189] The intermediate b (1 eq) was added to a reaction bottle, dissolved with dichloromethane (10 ml), and then DMAP (1.05 eq) and di-tert-butyl dicarbonate (1.5 eq) were added in sequence. After the addition was completed, the reaction was stirred at room temperature for 1 hour. After TLC monitoring, the reaction was completed, and a small amount of thin layer chromatography silica gel powder was added, and the mixture was concentrated under reduced pressure. After being spin-dried, the intermediate c was obtained by column chromatography (pure DCM: CH3OH=80:1, v / v), with a yield of about 80%.

[0190] Place intermediate c (1 eq) in a reaction bottle, dissolve it with DMF, and then add potassium carbonate (3 eq) and 3-bromopropylene (1.5 eq) in sequence. After stirring at room temperature for several hours, monitor by TLC that the reaction is complete, stop the reaction, add 10 times the volume of water as the solvent, extract with ethyl acetate (3×50 ml), concentrate the organic phase, add a small amount of thin layer chromatography silica gel powder, concentrate under reduced pressure, spin dry, and purify by column chromatography (PE:EA=(2:1, v / v) to obtain intermediate d (yield is about 60%).

[0191] The intermediate d (1 eq) was dissolved in dichloromethane (10 ml) in a reaction bottle, and trifluoroacetic acid (3 eq) was added dropwise at low temperature. After the addition, the mixture was reacted at room temperature for about one hour and monitored by TLC. After the reaction was completed, the reaction was stopped and the reaction solution was spin-dried. A small amount of water (30 ml) was added to dissolve the mixture, and the mixture was adjusted to weak alkalinity (pH = 8) with saturated sodium bicarbonate. The mixture was extracted with ethyl acetate (3×20 ml), and the organic phase was concentrated. A small amount of thin layer chromatography silica gel powder was added, and the mixture was concentrated under reduced pressure and spin-dried to obtain the intermediate e in a yield of about 50%.

[0192] The intermediate e (1 eq) was dissolved in ethanol and water (volume ratio = 3:1), and then sodium hydroxide (3 eq) was added. After reacting at room temperature for 2 hours, the reaction was stopped. The reaction solution was adjusted to slightly acidic (pH = 6), dried by spin drying, and chromatographed on a silica gel column (DCM: CH3OH = 30:1, v / v). Compound 53 (yield 60%) was obtained.

[0193] Compound 53: 1 H NMR(400MHz,DMSO-d6)δ8.88(s br,3H),7.03(d,J=8.4Hz,2H),6.66(d,J=8.4Hz,2H),5.89-5.76(m,1H),5.25(d,J=1 6.8Hz,1H),5.12(d,J=10.4Hz,1H),4.32(s,2H),3.82(d,J=5.2Hz,2H)ppm.HRMS:m / z calcd for C 14 H 14 N2O3S[M+H] + 291.0798, found 291.0791.

[0194] 2. Synthesis of compounds 56, 61, and 63

[0195] According to the synthesis method described for compound 53, the raw material 3-bromopropylene was replaced with 4-bromo-1-butene to prepare compound 56, the raw material 3-bromopropylene was replaced with benzyl bromide to prepare compound 61, and the intermediate b was directly hydrolyzed to prepare compound 63. The structures of each compound are shown below:

[0196]

[0197] The characterization data of the compounds are shown below:

[0198] Compound 56: 1 H NMR (400MHz, DMSO-d6) δ9.25(s,2H),7.03(d,J=6.4Hz,2H),6.67(d,J=8.0Hz,2H),5.87-5.77(m ,1H),5.11-5.01(m,2H),4.39-4.22(m,2H),3.24-3.17(m,2H),2.28-2.26(m,2H)ppm.HRMS:m / z calcd for C 15 H 16 N2O3S[M+H] + 305.0954, found 305.0947.

[0199] Compound 61: 1 H NMR (400MHz, DMSO-d6) δ7.14-7.09(m,4H),7.37-7.26(m,5H),7.05(d,J=8.4Hz,2H),6.69(d,J=8.4Hz,2H),4.50(s,2H),4.23(s,2H)ppm.HRMS:m / z calcdfor C 18 H 16 N2O3S[M+H] + 341.0954, found 341.0944.

[0200] Compound 63: 1 H NMR (400MHz, DMSO-d6) δ9.53(s br,1H),8.94(s br,1H),7.09(d,J=8.4Hz,2H),6.74(d,J=8.4Hz,2H),4.24(s,2H)ppm.HRMS:m / z calcd for C 11 H 10 N2O3S[M+H] +251.0485, found 251.0482.

[0201] Example 6. Synthesis of Compounds 59 and 70

[0202] Compounds 59 and 70 were synthesized by Suzuki reaction using p-bromothiazolamide as a substrate intermediate.

[0203] 1. Synthesis of compound 59

[0204] Synthesis route:

[0205]

[0206] Place p-bromothiazolamide (1 eq), phenylboronic acid (3 eq), potassium carbonate (3 eq), and bistriphenylphosphine palladium dichloride (0.1 eq) in a reaction flask, dissolve with toluene (15 ml), and reflux at 100° C. under Ar2 protection for several hours. Monitor by TLC. After the reaction is completed, stop the reaction, spin dry the solvent, add water (50 ml) to dissolve, extract with ethyl acetate (3×40 ml), dry the organic phase with anhydrous sodium sulfate, concentrate under reduced pressure and spin dry column chromatography PE:EA=(2:1, v / v) to obtain a yellow solid intermediate a (yield is about 80%).

[0207] Dissolve the intermediate a (1 eq) in ethanol and water (volume ratio = 3:1) in a total volume (12 ml), add sodium hydroxide (3 eq), react at room temperature for 2 hours, and then stop the reaction. Adjust the reaction solution to slightly acidic (pH = 6), spin dry, and perform silica gel column chromatography (DCM: CH3OH = 30:1, v / v). Compound 59 (yield 60%) was obtained.

[0208] Compound 59: 1 H NMR (400MHz, DMSO-d6) δ7.58-7.54(m,4H),7.32-7.30(m,4H),7.01(s,2H),4.37(s,2H),2.93-2.90(m,1H),1.23(d,J=6.8Hz,6H)ppm. 13 C NMR (101MHz, DMSO-d6) δ165.08,163.92,147.97,139.64,138.86,137.96,137. 04,136.08,129.28,127.30,127.09,126.97,33.55,32.13,24.32ppm.HRMS:m / z calcd for C 20 H 20 N2O2S[M+H] + 353.1318, found 353.1317.

[0209] 2. Synthesis of Compound 70

[0210] Synthesis route:

[0211]

[0212] Add p-bromothiazolamide (1 eq) to a reaction bottle, dissolve it with dichloromethane (20 ml), then add DMAP (1.05 eq) and di-tert-butyl dicarbonate (1.5 eq) in sequence. After the addition is complete, stir and react at room temperature for 1 hour. Monitor by TLC. After the reaction is complete, stop the reaction, add a small amount of thin layer chromatography silica gel powder, concentrate under reduced pressure, spin dry, and perform column chromatography on PE:EA=10:1, 8:1 (v / v) to obtain intermediate a with a yield of about 40%.

[0213] Place intermediate a (1 eq), phenylboronic acid (3 eq), cesium carbonate (3 eq), and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (0.3 eq) in a reaction flask, dissolve with 1,4-dioxane (15 ml), and reflux at 100° C. under Ar2 protection for several hours. Monitor by TLC. After the reaction is complete, stop the reaction, spin dry the solvent, add water (50 ml) to dissolve, extract with ethyl acetate (3x40 ml), dry the organic phase with anhydrous sodium sulfate, concentrate under reduced pressure and spin dry on a column chromatography PE:EA=(8:1, 5:1, 3:1, 2:1, v / v) to obtain a yellow solid intermediate b (yield is about 50%).

[0214] The intermediate b (1 eq) was dissolved in dichloromethane (15 ml) in a reaction bottle, and trifluoroacetic acid (3 eq) was added dropwise at low temperature. After the addition, the mixture was reacted at room temperature for about one hour and monitored by TLC. After the reaction was completed, the reaction was stopped and the reaction solution was spin-dried. A small amount of water (30 ml) was added to dissolve the mixture, and the mixture was adjusted to a weak alkaline state (pH = 8) with saturated sodium bicarbonate. The mixture was extracted with ethyl acetate (3 × 20 ml), and the organic phase was concentrated. A small amount of thin layer chromatography silica gel powder was added and the mixture was concentrated under reduced pressure. The intermediate c was obtained by column chromatography (pure DCM: CH3OH = 50:1, v / v) with a yield of about 70%.

[0215] Dissolve the intermediate c (1 eq) in a total volume (12 ml) of ethanol and water (volume ratio 3:1), add sodium hydroxide (3 eq), react at room temperature for 2 hours, and then stop the reaction. Adjust the reaction solution to slightly acidic (pH = 6), spin dry, and perform silica gel column chromatography (DCM: CH3OH = 30:1, v / v). Compound 70 (yield 60%) was obtained.

[0216] Compound 70: 1H NMR (400MHz, DMSO-d6) δ8.64(d,J=6.0Hz,2H),7.78-7.73(m,4H),7.39(d,J=8.4Hz,2H),7.05(s,2H),4.41(s,2H)ppm.

[0217] Example 7. Synthesis of Compounds 35-38

[0218] Synthesis route of compounds 35-38:

[0219]

[0220] R=CH3,Br,CH3O,C(CH3)3

[0221] Synthesis of Compounds 35-38

[0222] The raw materials p-methylphenethanol (204 mg, 1.50 mmol), p-bromophenethanol (100 mg, 0.50 mmol), p-methoxyphenethanol (200 mg, 1.31 mmol) and p-tert-butylphenethanol (200 mg, 1.12 mmol) were placed in a reaction bottle, dissolved in dichloromethane (15 mL), and Dess-Martin (764 mg, 1.80 mmol), (253 mg, 0.60 mmol), (669 mg, 1.58 mmol) and (517 mg, 1.35 mmol) were added respectively. The temperature was raised to 40°C for reflux reaction. After 2 h, TLC spot plate detection was performed. After the reaction was complete, the dichloromethane was dried by spin drying, and saturated sodium thiosulfate aqueous solution and ethyl acetate were added for extraction. The extract was concentrated and subjected to column chromatography (PE→PE:EA=30:1, v / v) to obtain 40 mg of colorless liquid compound a (p-methyl) with a yield of 20%; and light yellow liquid compound b 47 mg (p-bromine), yield 47%; colorless liquid compound c 110 mg (p-methoxy), yield 55.8%; and colorless liquid compound d 92 mg (p-tert-butyl), yield 46.3%.

[0223] Compound a (350 mg, 2.61 mmol), b (100 mg, 0.50 mmol), c (110 mg, 0.73 mmol), d (200 mg, 1.10 mmol) were placed in reaction bottles, dissolved in anhydrous ether (15 mL), and then methyl dichloroacetate (338 μL, 3.26 mmol), (65 μL, 0.63 mmol), (99 μL, 0.95 mmol), (150 μL, 1.50 mmol) were added. After precooling at 0 °C, 30% sodium methoxide (497 μL, 3.59 mmol), (86 μL, 0.69 mmol), (152 μL, 1.10 mmole) were slowly added dropwise. ol), (240μL, 1.72mmol), react for 1h and then move to room temperature to react for 3h, then detect by TLC, after the reaction is complete, extract with saturated sodium chloride solution and anhydrous ether respectively, concentrate the extract and add methanol (10mL) to dissolve, then add thiourea (198mg, 2.61mmol), (38mg, 0.50mmol), c (56mg, 0.73mmol), (90mg, 1.13mmol) respectively, heat to 65℃ for reflux reaction, detect by TLC after 3h, after the reaction is complete, concentrate and purify by column chromatography (PE:EA=5:1→PE:EA=1:1, v / v) to obtain pale yellow solid compound e respectively. 70mg (p-methyl), yield 10.3%; pale yellow solid compound f 120mg (p-bromine), yield 73.3%; pale yellow solid compound g 131mg (p-methoxy), yield 68.1% and yellow viscous liquid compound h 120mg (p-tert-butyl), yield 36.5%.

[0224] Compounds e (70 mg, 0.27 mmol), f (120 mg, 0.37 mmol), g (131 mg, 0.47 mmol), and h (120 mg, 0.39 mmol) were dissolved in EtOH:H2O (6:3 mL), and sodium hydroxide (22 mg, 0.80 mmol), (44 mg, 1.11 mmol), (56 mg, 1.41 mmol), and (47 mg, 1.17 mmol) were added respectively, and the temperature was raised to 65°C for reflux reaction. After 1 h, TLC spot plate detection was performed. After the reaction was complete, ethanol was spin-dried and 2M HCl was used to adjust the pH to about 7, and a white solid was precipitated. After filtering, the solid was washed out with dichloromethane:methanol=20:1 (v / v) to obtain white compound 35 (28 mg, yield 42%); white solid compound 36 (60 mg, yield 52%); solid compound 38 (55 mg, yield 44%); solid compound 37 (52 mg, yield 46%). The structures of each compound are shown below:

[0225]

[0226] The characterization data of each compound are shown below:

[0227] Compound 35: 1 H NMR (400MHz, DMSO-d6) δ7.12 (dd, J=13.2Hz, J=7.2Hz, 4H), 4.40 (s, 2H), 2.25 (s, 3H) ppm. HRMS: m / z calcd for C 12 H 12 N2O2S[MH] - 247.0546, found 247.0560.

[0228] Compound 36: 1 H NMR (400MHz, DMSO-d6) δ7.46 (d, J = 8Hz 2H), 7.21 (d, J = 8Hz, 2H), 6.94 (s, 2H), 4.38 (s, 2H) ppm.

[0229] Compound 37: 1 H NMR (400MHz, DMSO-d6) δ7.31(d,J=8.0Hz,2H),7.15(d,J=8.0Hz,2H),6.90(s,2H),4.28(s,2H),1.25(s,9H)ppm.

[0230] Compound 38: 1 H NMR (400MHz, DMSO-d6) δ7.16(d,J=8.0Hz,2H),6.95(s,1H),6.86(d,J=8.4Hz,2H),4.26(s,2H),3.72(s,3H)ppm.

[0231] Example 8. Synthesis of Compounds 3, 4, 6, 9, 10, 15, 19, 23, 25, 30, 32, 33 and 34

[0232]

[0233] Dissolve intermediate a (400 mg, 1.6 mmol, 1 eq) in hypophosphorous acid, slowly add sodium nitrite (222.3 mg, 3.2 mmol, 2 eq) in water under ice bath, react at 0°C for 1 hour and then move to room temperature for 2 hours, stop the reaction, dilute with water, adjust to weak base with sodium bicarbonate, extract with ethyl acetate, dry over anhydrous sodium sulfate, concentrate under reduced pressure, and perform silica gel column chromatography (PE:EA=8:1, 6:1, 4:1, 2:1, v / v) to obtain intermediate b (230 mg, yield 58%).

[0234] Dissolve intermediate b (200 mg, 0.86 mmol, 1 eq) in ethanol and water, add sodium hydroxide (103 mg, 2.57 mmol, 3 eq), react at room temperature for 2 hours, and then stop the reaction. Adjust the reaction solution to slightly acidic, spin dry, and perform silica gel column chromatography (DCM: CH3OH = 50:1, 30; 1, v / v). Compound 3 (160 mg, yield 80%) was obtained.

[0235] Compound 3: 1 H NMR (400MHz, DMSO-d6) δ13.10(s,1H),8.91(s,1H),7.34-7.24(m,5H),4.57(s,2H)ppm. 13 C NMR(101MHz,DMSO-d6)δ163.91,152.10,149.05,142.51,140.28,129.15,128.99,127.25,32.78ppm.HRMS:m / z calcd for C 11 H9NO2S[M+H] + 220.0427,found220.0429; [M-H2O+H] + 202.0327, found 202.0317.

[0236]

[0237] The raw material (200 mg, 0.76 mmol, 1 eq) was dissolved in acetonitrile, and tert-butyl nitrite (452 ​​μl, 3.8 mmol, 5 eq) was added dropwise at 0°C. After reacting for 20 min, copper chloride (260 mg, 1.52 mmol, 2 eq) was added and the mixture was moved to room temperature for reaction. After 12 hours, the reaction was stopped, the acetonitrile was dried by spin drying, and extracted with water and ethyl acetate, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to silica gel column chromatography (PE: EA = 20: 1, v / v) to obtain intermediate a (120 mg, yield 60%).

[0238] Dissolve intermediate a (100 mg, 0.36 mmol, 1 eq) in ethanol and water, add sodium hydroxide (43 mg, 1.06 mmol, 3 eq), react at room temperature for 2 hours, and then stop the reaction. Adjust the reaction solution to slightly acidic, spin dry, and perform silica gel column chromatography (DCM: CH3OH=50:1, 30:1, v / v). Solid compound 9 (73 mg, yield 73%) was obtained.

[0239] Compound 9: 1H NMR (400MHz, DMSO-d6) δ13.27(s,1H),7.32-7.28(m,2H),7.23-7.21(m,3H),3.47(t,J=8.0,Hz,2H),2.92(t,J=8.0,Hz,2H)ppm.HRMS:m / z calcd forC 12 H 10 ClNO2S[MH] - 266.0048, found 266.0015.

[0240] The synthesis methods of compounds 4, 6, 19, 23, 25, 30 and 32 are the same as compound 3.

[0241] The synthesis methods of compounds 10, 15, 33 and 34 are the same as compound 9.

[0242]

[0243]

[0244] The characterization data of each compound are shown below:

[0245] Compound 4: 1 H NMR (400MHz, DMSO-d6) δ13.08(s,1H),8.94(s,1H),7.32(d,J=4.4Hz,4H),7.26-7.20(m,1H),5.37(q,J=7.2Hz,1H),1.64(d,J=7.2Hz,3H)ppm. 13 C NMR(101MHz,DMSO-d6)δ163.85,155.33,151.87,145.12,141.79,129.07,127.45,127.22,37.14,23.79ppm.HRMS m / z calcd for C 12 H 11 NO2S[M+H] + 234.0583, found 234.0583; [M-H2O+H] + 216.0483,found216.0470;[M+Na] + 256.0408, found 256.0401.

[0246] Compound 6: 11H NMR (400 MHz, DMSO-d6) δ 12.95 (s, 1H), 8.88 (s, 1H), 7.31 - 7.18 (m, 5H), 3.50 (t, J = 8.0 Hz, 2H), 2.94 (t, J = 8.0 Hz, 2H) ppm. 13 13C NMR (101 MHz, DMSO-d6) δ 163.78, 151.28, 148.65, 142.69, 140.83, 128.88, 128.82, 126.68, 37.26, 28.93 ppm. HRMS: m / z calcd for C 12 H 11 NO2S [M + H] + 234.0583, found 234.0599; [M - H2O + H] + 216.0483, found 216.0477; [M + Na] + 256.0408, found 256.0408.

[0247] Compound 10: 1 1H NMR (400 MHz, DMSO-d6) δ 13.25 (s, 1H), 7.31 - 7.28 (m, 2H), 7.21 - 7.19 (m, 3H), 3.47 (t, J = 8.0 Hz, 2H), 2.92 (t, J = 8.0 Hz, 2H) ppm. HRMS: m / z calcd for C 12 H 10 BrNO2S [M + H] + 311.9688, found 311.9679, 313.9659; [M - H2O + H] + 293.9588, found 293.9570, 295.9550.

[0248] Compound 15: HRMS: m / z calcd for C 12 H 10 INO2S [M + H] + 359.9550, found 359.9554; C 12 H 10 INO2S [M + H] + 360.9583, found 360.9595; C 12 H 10 INO2S [M + Na] + 381.9369, found 381.9363; C 12 H 10INO2S[M+Na] + 382.9403, found 3882.9408.

[0249] Compound 19: 1 H NMR (400MHz, DMSO-d6) δ13.16(s,1H),9.21(s,1H),8.28(dd,J=6.8,J=1.6,2H),7.83(d,J=8.4,2H)ppm.HRMS:m / z calcd for C 10 H6N2O4S[MH] - 248.9976, found 248.9973.

[0250] Compound 23: 1 H NMR (400MHz, CDCl3) δ8.81 (s, 1H), 4.33 (s, 1H), 1.25 (d, J = 16.4, 6H) ppm. HRMS: m / z calcd for C 10 H9NO2S[M+H] + 172.0427, found 172.0432; [M-H2O+H] + 154.0327, found 154.0323.

[0251] Compound 25: LC-MS m / z: 186.1 [M+H]+.

[0252] Compound 30: 1 H NMR (400MHz, DMSO-d6) δ8.69(s,1H),3.18(s,2H),1.86(s,1H),0.88(s,6H)ppm.

[0253] Compound 32: 1 H NMR(400MHz,DMSO-d6)δ9.17(s,1H),8.52(s,1H)ppm.HRMS:m / zcalcd for C4H3NO2S[MH] - 127.9812,found 127.9915,[M-H2O] - 110.9778, found110.9760.

[0254] Compound 33: 1 H NMR(400MHz,DMSO-d6)δ7.30-7.24(m,5H),4.61(s,2H)ppm.HRMS:m / zcalcd for C 11H8ClNO2S[MH] - 251.9892, found 251.9854.

[0255] Compound 34: 1 H NMR(400MHz,DMSO-d6)δ7.29-7.23(m,5H),4.62(s,2H)ppm.HRMS:m / zcalcd for C 11 H8BrNO2S[MH] - 295.9386,found 295.9349,297.9327.

[0256] Example 9, Synthesis of Compound 7

[0257]

[0258] Dissolve 2-amino-5-bromoformic acid ethyl ester (300 mg, 1.19 mmol, 1 eq) in 5 ml of toluene, add thiophene boronic acid (456 mg, 3.58 mmol, 3 eq), cesium carbonate (1.17 g, 3.58 mmol, 3 eq), Pd(pph3)4 (138 mg, 0.119 mmol, 0.1 eq), and then heat to 110°C and reflux for 5 hours. Stop the reaction, spin dry toluene, extract with water and ethyl acetate, dry over anhydrous sodium sulfate, concentrate under reduced pressure, and perform silica gel column chromatography (PE:EA=8:1, 6:1, 4:1, 2:1, v / v) to obtain intermediate a (210 mg, yield 70%).

[0259] Dissolve intermediate a (200 mg, 0.83 mmol, 1 eq) in ethanol and water, add sodium hydroxide (100 mg, 2.5 mmol, 3 eq), react at room temperature for 2 hours, and then stop the reaction. Adjust the reaction solution to slightly acidic, spin dry, and perform silica gel column chromatography (DCM: CH3OH = 50:1, 30:1, 10:1, v / v). Obtain compound 7 (40 mg, yield 20%).

[0260] Compound 7: 1 H NMR (400MHz, DMSO-d6) δ8.56(s br,2H),7.89(d,J=2.0Hz,1H),7.63-7.61(m,1H),7.36(dd,J=4.8Hz,J=0.8Hz,1H)ppm.HRMS:m / z calcd for C8H6N2O2S2[M+H] + 226.9943, found 226.9945; [M-H2O+H] +208.9843, found 258.9767; [M+Na] + 248.9768, found 258.9767.

[0261] Example 10. Synthesis of Compounds 8 and 16

[0262]

[0263] Sodium flakes (69mh, 2.9mmol, 1.5eq) were added to ethanol. After complete dissolution, 2-amino-5-ethylbromoformate (500mg, 1.99mmol, 1eq) and p-chlorothiophenol (230mg, 1.6mmol, 0.8eq) were added. The mixture was moved to 80°C for reaction for two hours. The reaction was stopped, filtered, and the filtrate was dried by rotary evaporation. After dissolving in ethyl acetate, the yellow solid intermediate a (230mg, yield 46%) was obtained by silica gel column chromatography (PE:EA=7:1, 5:1, 3:1, 1:1, v / v).

[0264] Dissolve intermediate a (120 mg, 0.38 mmol, 1 eq) in ethanol and water, add sodium hydroxide (46 mg, 1.14 mmol, 3 eq), react at room temperature for 2 hours, and then stop the reaction. Adjust the reaction solution to slightly acidic, spin dry, and perform silica gel column chromatography (DCM: CH3OH = 50:1, 30:1, 10:1). Obtain red solid compound 8 (80 mg, yield 76%).

[0265] Compound 8: 1 H NMR(400MHz,DMSO-d6)δ7.45-7.38(m,6H)ppm.HRMS:m / z calcd forC 10 H7ClN2O2S2[M+H] + 286.9710, found 286.9700; [M-H2O+H] + 268.9610, found 268.9590.

[0266] After dissolving intermediate a (200 mg, 0.64 mmol, 1 eq) in dry dichloromethane, MCPBA (328 mg, 1.9 mmol, 3 eq) was added at 0°C. After stirring at room temperature for 6 hours, the reaction was stopped and filtered. The filtrate was subjected to silica gel column chromatography (DCM:CH3OH=40:1) to obtain light yellow solid intermediate b (130 mg, yield 65%).

[0267] Dissolve intermediate b (100 mg, 0.29 mmol, 1 eq) in ethanol and water, add sodium hydroxide (35 mg, 0.87 mmol, 3 eq), react at room temperature for 2 hours, and then stop the reaction. Adjust the reaction solution to slightly acidic, spin dry, and perform silica gel column chromatography (DCM: CH3OH=50:1, 30:1, v / v). Obtain yellow solid compound 16 (37 mg, yield 37%).

[0268] Compound 16: 1 H NMR (400MHz, DMSO-d6) δ13.75 (s, 1H), 8.23 ​​(s, 2H), 7.94 (d, J = 8.4Hz, 2H), 7.72 (d, J = 8.4Hz, 2H) ppm.HRMS: m / z calcd for C 10 H7ClN2O4S2[M+H] + 318.9609, found 318.9607; [M-H2O+H] + 300.9509, found 300.9509; [M+Na] + 340.9433, found 340.9436.

[0269] Example 11. Synthesis of Compound 27

[0270]

[0271] Accurately weigh the compound 2-amino-5-(sec-butyl)thiazole-4-carboxylic acid methyl ester 7b (200 mg, 0.9 mmol) and dissolve it in 8 ml of dry DCM. Add triethylamine (283 mg, 2.8 mmol) and acetyl chloride (110 mg, 1.4 mmol) under ice bath. After the addition, warm to room temperature for reaction. After 3 h, TLC detection showed that the reaction was complete. The reaction was stopped and concentrated under reduced pressure to remove DCM. Purification by column chromatography (PE: EA = 4: 1) gave an off-white solid, i.e., compound 2-acetylamino-5-(sec-butyl)thiazole-4-carboxylic acid methyl ester 9b (190 mg, yield 79.5%).

[0272] Using compound 2-acetylamino-5-(sec-butyl)thiazole-4-carboxylic acid methyl ester 9b as starting material, according to the general method of ester hydrolysis, it was purified by column chromatography (DCM:MeOH=10:1) to obtain a white solid, namely compound 2-acetylamino-5-(sec-butyl)thiazole-4-carboxylic acid 9 (compound 27, 82 mg, yield 86.7%).

[0273] Compound 27: HRMS: m / z calcd for C 10 H 14 N2O3S[M+H] +243.0798, found 243.0814; [M-H2O+H] + 225.0698, found 225.0708; [M+Na] + 265.0623, found 265.0622.

[0274] Compound 31 was synthesized according to the synthetic method of compound 27 by replacing acetyl chloride with benzoyl chloride.

[0275]

[0276] Compound 31: 1 H NMR (400MHz, DMSO-d6) δ7.41-7.22(m,5H),4.07(s,2H),2.92(s,2H),1.69(s,1H),0.83(d,J=3.2Hz,6H)ppm.HRMS:m / z calcd for C 15 H 18 N2O2S[M+H] + 291.1162, found 291.1152; [M-H2O+H] + 273.1062, found 273.1054; [M+Na] + 313.0987, found 313.0979.

[0277] The beneficial effects of the present invention are demonstrated below through specific test examples.

[0278] Test Example 1: Inhibitory activity of the compounds of the present invention on MBL

[0279] 1. Test methods

[0280] The activity test reaction was carried out in a black 96-well ELISA plate with a total system of 60 μL. The specific operation steps are as follows:

[0281] (1) The solid compound to be tested was prepared into a 100 mM stock solution with DMSO, and the stock solution was diluted into a 3.6 mM or 600 μM working solution with test buffer (20 mM Tris-HCl pH 7.5, 200 mM NaCl, 0.01% Triton X-100). The working solution was then diluted three-fold with test buffer into 9 concentration gradients to obtain 10 working solutions of different concentrations.

[0282] (2) Prepare the MBL enzyme solution with a test buffer to a certain concentration (VIM-2, VIM-1 and VIM-5: 1.2 nM; NDM-1: 0.5 nM; IMP-1: 4.8 nM), and prepare the substrate FC-5 with a test buffer to a 30 μM substrate solution for later use.

[0283] (3) In a 96-well ELISA plate, 10 μL of the compound working solution obtained in step (1), 30 μL of the test buffer, and 10 μL of the enzyme solution were added to each well in sequence and incubated at room temperature for 10 min.

[0284] (4) Add 10 μL of substrate solution to each well of a 96-well ELISA plate and use a Tecan ELISA reader to continuously measure the fluorescence value (λ ex =380nm,λ em =460nm) changes.

[0285] Each experiment set up a reaction well without adding compound as a control, and all measurements included three sets of parallel experiments. According to the change of fluorescence intensity measured by the microplate reader, the residual activity of the enzyme in each well was calculated, and the calculation was expressed as: residual activity (%) = (ΔF I ) / (ΔF C )×100,ΔF I ΔF is the fluorescence change value of the well containing the compound within a certain period of time. C is the fluorescence change value in the control well without adding the compound in the same time period. The processed data were fitted using Graphpad Prism 5 software to obtain IC 50 value.

[0286] 2. Test results

[0287] Table 1. Inhibitory activity of each compound on MBL a

[0288]

[0289]

[0290]

[0291] a ****: indicates good inhibitory activity (IC 50 <1μM; ***: represents moderate inhibitory activity (1μM) <IC 50 <100μM; **: represents weak inhibitory activity (IC 50 >100μM)

[0292] The test results show that the compounds of the present invention have good inhibitory activity against MBL, especially against multiple MBL subtypes, especially against clinically relevant subtypes such as VIM, NDM, IMP, etc., among which compound 64 is the best.

[0293] Test Example 2: Antibacterial activity of the compound of the present invention in combination with meropenem

[0294] 1. Test methods

[0295] (1) Resuscitation of strains: Resuscitate the strains from a -80°C freezer, streak the strains onto non-resistant MHA plates using an inoculation loop, and culture at 35°C for 18-20 h.

[0296] (2) Drug dilution: Weigh meropenem solid and dissolve it in DMSO to make a 12.8 mg / ml stock solution. Dilute the stock solution with CAMHB medium to a 512 μg / mL working solution. Add 100 μL CAMHB to a transparent 96-well plate, then add 100 μL meropenem working solution, then dilute 9 concentrations by 2 times, and set up a growth control well without meropenem. Add 2 μL of the compound at a concentration of 1 mg / mL to each well.

[0297] (3) Bacterial dilution: Use an inoculation loop to pick a single clone from the plate and place it in saline. Adjust the OD 630 to 0.08-0.13 (equivalent to 1×10 8 CFU / mL), dilute the bacterial solution 100 times with CAMHB, and add 100 μL to each well. At this time, the concentration of meropenem is 128 μg / mL to 0.25 μg / mL, and the concentration of enzyme inhibitor is 10 μg / mL.

[0298] (4) Static culture and result determination: Place the 96-well plate in a 35°C incubator and incubate for 16-20 hours. Observe the results the next day, and the minimum concentration that can completely or significantly inhibit bacterial growth is the minimum inhibitory concentration (MIC) of meropenem.

[0299] 2. Test results

[0300] Table 2. Antibacterial activity of each compound in combination with meropenem against clinical isolates expressing MBL

[0301]

[0302]

[0303] The above test results indicate that most of the compounds of the present invention, such as compounds 1, 2, 21, 54, 64, etc., have good antibacterial activity against clinically isolated strains such as Escherichia coli and Klebsiella pneumoniae expressing MBL when used in combination with meropenem.

[0304] In summary, the compounds of the present invention have good inhibitory activity against metallo-β-lactamase (MBL), especially against multiple clinically relevant MBL subtypes, such as VIM, NDM, IMP and other subtypes, have strong Gram-negative bacteria permeability, can be used to prepare broad-spectrum MBL inhibitors, and have good development prospects as drug molecules to overcome antimicrobial resistance.

Claims

1. The compound shown below, or its salt:

2. A drug, characterized in that: The preparation is prepared by taking the compound described in claim 1 or its salt as active ingredient and adding pharmaceutically acceptable auxiliary materials or auxiliary ingredients.

3. Use of the compound of formula I or its salt in the preparation of MBL inhibitors and / or the compound of formula I or its salt in combination with meropenem in the preparation of antibacterial drugs: R1 is selected from halogen, amino, -NHR1'; R1' is selected from C1-C3 alkyl substituted by 0-1 R3, -C(O)R3'; R3 are independently selected from 5-10 membered aryl and 5-10 membered heteroaryl; R3' is selected from C1-C3 alkyl; L is selected from C1-C6 alkylene substituted with 0-1 R4, -O-, R4 are independently selected from C1 to C3 alkyl; R2 is selected from C1-C7 alkyl, C2-C4 alkenyl, C2-C4 alkynyl, phenyl substituted by 0-2 R5, thienyl substituted by 0-1 R5, piperidinyl substituted by 0-1 R5, azetidine substituted by 0-1 R5, naphthyl substituted by 0-1 R5; R5 is independently selected from C1-C4 alkyl substituted by 0-3 R6, C1-C3 alkoxy substituted by 0-3 R6, carboxyl, nitro, amino, hydroxyl, -C(O)R7, -C(O)OR7, phenyl substituted by 0-1 R6, pyridyl substituted by 0-1 R6; R6 are independently selected from C1-C3 alkyl and halogen; R7 is selected from C1-C4 alkyl; When R1 is selected from amino, R2 is selected from C1 alkyl, L is not selected from C2-C4 alkylene, none, C2 alkylene substituted with 1 C1 alkyl, C1 alkylene substituted with 1 C1 alkyl; When R1 is selected from amino and R2 is selected from phenyl, L is not selected from C1 alkylene; When R1 is selected from amino, L is selected from C1 alkylene, R is not selected from 4. The use according to claim 3, characterized in that: The compound is shown in Formula II: in, L is selected from C1-C6 alkylene substituted with 0-1 R4, -O-, R4 are independently selected from C1 to C3 alkyl; R2 is selected from C1-C7 alkyl, C2-C4 alkenyl, C2-C4 alkynyl, phenyl substituted by 0-2 R5, thienyl substituted by 0-1 R5, piperidinyl substituted by 0-1 R5, azetidine substituted by 0-1 R5, naphthyl substituted by 0-1 R5; R5 is independently selected from C1-C4 alkyl substituted by 0-3 R6, C1-C3 alkoxy substituted by 0-3 R6, carboxyl, nitro, amino, hydroxyl, -C(O)R7, -C(O)OR7, phenyl substituted by 0-1 R6, pyridyl substituted by 0-1 R6; R6 are independently selected from C1-C3 alkyl and halogen; R7 is selected from C1-C4 alkyl.

5. The use according to claim 3, characterized in that: The compound is shown in formula III: in, R1 is selected from halogen, amino, -NHR1'; R1' is selected from C1-C3 alkyl substituted by 0-1 R3, -C(O)R3'; R3 are independently selected from phenyl, pyridyl, thienyl, pyrazolyl, furyl; R3' is selected from C1-C3 alkyl; L is selected from C1-C6 alkylene substituted with 0-1 R4, -O-, R4 are independently selected from C1 to C3 alkyl; R 5a , R 5b , R 5c Each of the following groups is independently selected from hydrogen, C1-C4 alkyl substituted by 0-3 R6, C1-C3 alkoxy substituted by 0-3 R6, carboxyl, nitro, amino, hydroxyl, -C(O)R7, -C(O)OR7, phenyl substituted by 0-1 R6, and pyridyl substituted by 0-1 R6; R6 are independently selected from C1-C3 alkyl and halogen; R7 is selected from C1-C4 alkyl.

6. The use according to claim 4, characterized in that: The compound is shown in Formula IV: in, R1 is selected from halogen, amino, -NHR1'; R1' is selected from C1-C3 alkyl substituted by 0-1 R3, -C(O)R3'; R3 are independently selected from phenyl, pyridyl, thienyl, pyrazolyl, furyl; R3' is selected from C1-C3 alkyl; L is selected from C1-C6 alkylene substituted with 0-1 R4, -O-, R4 are each independently selected from C1 to C3 alkyl.

7. Use of the following compounds or their salts in the preparation of MBL inhibitors and / or the following compounds or their salts in combination with meropenem in the preparation of antibacterial drugs:

8. Use of a thiazolamide compound or a salt thereof in the preparation of an MBL inhibitor and / or in combination with meropenem in the preparation of an antibacterial drug, characterized in that: The thiazole amine compound is one of the following compounds:

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