Thiazole amine-diazabicyclooctanone conjugate derivatives and uses thereof

Abstract

The application provides a kind of thiazole amine-diazabicyclo octanone conjugate derivative and its use, belong to the field of medicine.The compound provided by the application has good, broad-spectrum inhibitory activity on metal beta lactamase (MBL) and serine beta lactamase (SBL), and can be used for preparing MBL and / or SBL inhibitor.The compound of the application has great potential in preparing MBL and / or SBL inhibitor and antibacterial drug (especially drug for resisting drug-resistant bacteria).

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of medicine, and particularly relates to a class of thiazole amine-diazabicyclooctanone conjugate derivatives and uses thereof. BACKGROUND

[0002] β-lactam antibiotics are the most widely used antibiotics in clinical, such as penicillins, cephalosporins, carbapenems, monobactams, etc. Among them, carbapenem antibiotics are the most widely used and the strongest bactericidal class of β-lactam antibiotics, and are one of the most important drugs for the treatment of serious bacterial infections in clinical, and have been hailed as the "last line of defense" against bacterial infections. However, β-lactam antibiotic resistance is very common, and the drug resistance situation is very serious. In the list of drug-resistant bacteria announced by the World Health Organization in 2017, carbapenem-resistant Acinetobacter baumannii, Pseudomonas aeruginosa and Enterobacteriaceae bacteria are listed as the most dangerous superbugs, and there is still a lack of effective treatment drugs, which has attracted worldwide attention and vigilance. Research has found that the most important mechanism of β-lactam antibiotics, including carbapenem, resistance is that the pathogenic bacteria themselves produce β-lactamase to hydrolyze the β-lactam ring core pharmacophore, thereby losing antibacterial activity.

[0003] According to the Am-bler classification method, β-lactamase can be divided into A, B, C, D 4 types, among which the active center of A, C, D class is serine, collectively known as serine β-lactamase (SBL); the active center of B class contains at least one Zn 2+ Therefore, B class β-lactamase is also called metallo-β-lactamase (MBL). The combination of β-lactam antibiotics and β-lactamase inhibitors has become one of the main means to overcome antibiotic resistance in clinical. The β-lactamase inhibitors currently used in clinical mainly include clavulanic acid, sulbactam, tazobactam and newly listed avibactam (Avibactam). Clavulanic acid, sulbactam and tazobactam are irreversible inhibitors, which only have inhibitory effect on most A class β-lactamase, and have no inhibitory effect on B, C, D class β-lactamase. Avibactam is a reversible inhibitor, which has inhibitory effect on A class, C class and part of D class β-lactamase, but its inhibitory effect on B class β-lactamase is not good.

[0004]

[0005] Due to the genetic material exchange and transfer of the genes expressing drug resistance enzymes between the same or different pathogenic bacteria species, drug-resistant bacteria co-expressing MBL and SBL are constantly emerging and spreading, which poses a great threat to human life and health. In the face of the increasing number of multiple drug-resistant, extensively-drug resistant (XDR) and even pan-drug resistant (PDR)'superbacteria', the existing β-lactamase inhibitors cannot meet the clinical needs, and inhibitors with dual inhibition of MBL and SBL need to be developed urgently. SUMMARY

[0006] The present application aims to provide a class of thiazole amine-diazabicyclo octanone conjugate derivatives and their preparation method and use.

[0007] The present application provides a compound, its conformational isomer, its optical isomer or its pharmaceutically acceptable salt, the structure of the compound is shown in formula I:

[0008]

[0009] L is a linker or nothing.

[0010] Further, L is selected from C 1-8 alkylene,

[0011] X is selected from nothing, C 1-8 alkylene; Y is selected from nothing, C 1-8 alkylene;

[0012] A ring is selected from 3-6 membered saturated cycloalkyl, 3-6 membered saturated heterocyclyl, 5-6 membered aryl, 5-6 membered heteroaryl, fused cycloalkyl, heterofused cyclyl;

[0013] m is selected from 0, 1, 2, 3;

[0014] R1is each independently selected from hydrogen, C 1-8 alkyl, C 1-8 alkoxy, halogen, hydroxyl, amino, carboxyl, cyano.

[0015] Further, the structure of the compound is shown in formula II:

[0016]

[0017] n is selected from 0, 1, 2, 3, 4, 5.

[0018] Further, the structure of the compound is shown in formula III:

[0019]

[0020] wherein x1 is selected from 0, 1, 2, 3, 4, 5;

[0021] y1 is selected from 0, 1, 2, 3, 4, 5;

[0022] m is selected from 0, 1, 2, 3;

[0023] each R1 is independently selected from hydrogen, C 1-8 alkyl, C 1-8 alkoxy, halogen, hydroxyl, amino, carboxyl, cyano.

[0024] Further, the structure of the compound is shown in formula IV:

[0025]

[0026] wherein x2 is selected from 0, 1, 2, 3, 4, 5;

[0027] y2 is selected from 0, 1, 2, 3, 4, 5;

[0028] R1 is selected from hydrogen, C 1-8 alkyl, C 1-8 alkoxy, halogen, hydroxyl, amino, carboxyl, cyano.

[0029] Z is selected from O, S or CH2.

[0030] Further, the compound is selected from:

[0031]

[0032] The present application provides an antibacterial drug, which is prepared from the above compound, conformational isomer thereof, optical isomer thereof or pharmaceutically acceptable salt thereof as an active ingredient, plus a pharmaceutically acceptable adjuvant.

[0033] The present application also provides the use of the above compound, conformational isomer thereof, optical isomer thereof or pharmaceutically acceptable salt thereof in the preparation of a β-lactamase inhibitor.

[0034] Further, the β-lactamase inhibitor is a metallo-β-lactamase inhibitor, a serine β-lactamase inhibitor, a metallo-β-lactamase and serine β-lactamase dual inhibitor.

[0035] Further, the β-lactamase inhibitor is an antibacterial drug.

[0036] Further, the antibacterial drug is a drug against drug-resistant bacteria, preferably a drug against bacteria resistant to β-lactam antibiotics.

[0037] The present application also provides a combination drug with antibacterial efficacy, which comprises the above-mentioned compound, conformational isomer, optical isomer or pharmaceutically acceptable salt thereof and β-lactam antibiotic, and a pharmaceutically acceptable carrier.

[0038] Definitions of terms used in the present application: Unless otherwise indicated, the initial definition of a group or term provided herein applies throughout the specification; for terms not specifically defined herein, the meaning given such terms by one of ordinary skill in the art in light of the disclosure and context will prevail.

[0039] The minimum and maximum number of carbon atoms in a hydrocarbon group is indicated by a prefix, e.g., the prefix C a～b Alkyl refers to a straight or branched chain alkyl group containing 1 to 8 carbon atoms. 1～8 Alkyl refers to a straight or branched chain alkyl group containing 1 to 8 carbon atoms.

[0040] "Aryl" refers to a fully carbon monocyclic ring group having a conjugated pi electron system, e.g., phenyl. The aryl group does not contain heteroatoms, such as nitrogen, oxygen, or sulfur, and the point of attachment to the parent molecule must be through a carbon atom on the ring having the conjugated pi electron system.

[0041] "Heteroaryl" refers to a heteroaromatic group containing one to several heteroatoms. The heteroatoms referred to herein include oxygen, sulfur, and nitrogen. Examples include furyl, thienyl, pyridyl, pyrazolyl, pyrrolyl, N-alkyl pyrrolyl, pyrimidinyl, pyrazinyl, imidazolyl, tetrazolyl, and the like.

[0042] "Cycloalkyl" refers to a saturated or unsaturated cyclic alkyl group.

[0043] "Heterocycloalkyl" refers to a saturated or unsaturated cyclic heteroalkyl group.

[0044] Halogen is fluorine, chlorine, bromine or iodine.

[0045] Linker.

[0046] The compound provided by the present application has good and broad-spectrum inhibitory activity on metallo-beta-lactamase (MBL) and serine beta-lactamase (SBL), and can be used for preparing an inhibitor of MBL and / or SBL. The compound of the present application has great potential in preparing an inhibitor of MBL and / or SBL and an antibacterial drug (especially a drug against drug-resistant bacteria).

[0047] Obviously, according to the above content of the present application, according to the ordinary technical knowledge and means in the art, other various forms of modification, replacement or change can be made without departing from the above basic technical idea of the present application.

[0048] The above content of the present application will be further illustrated in detail by the following specific embodiments in the form of examples. However, it should not be understood that the scope of the above subject matter of the present application is limited to the following examples. Any technology achieved based on the above content of the present application belongs to the scope of the present application. DETAILED DESCRIPTION

[0049] The raw materials and equipment used in the present application are known products, which are obtained by purchasing commercially available products.

[0050] The following is the synthesis method of intermediates a4 and a7'.

[0051] 1. Synthesis of intermediate a4

[0052]

[0053] 2-Amino-5-bromo-4-thiazolecarboxylic acid ethyl ester (a1, 1.0 eq), di-tert-butyl dicarbonate (1.5 eq), 4-dimethylaminopyridine (0.2 eq) and triethylamine (3.0 eq) were added to dry dichloromethane (15 mL / 1.0 mmoL) respectively, and reacted at room temperature for 2 hours. TLC was used to monitor the complete reaction of the raw material 1, and column chromatography (petroleum ether: ethyl acetate = 10:1) was used for purification to obtain compound intermediate 5-bromo-2-(tert-butoxycarbonyl) amino) thiazole-4-carboxylic acid ethyl ester (a2, light yellow solid) with a yield of 92%. ESI-MS: m / z calcd for C 11 H 15 BrN2O4S[M+H] + 351.0, found 351.0, 353.0.

[0054] Intermediate a2 (1.0 eq) was dissolved in tetrahydrofuran solution (15 mL / 1.0 mmoL), and then 10% potassium hydroxide aqueous solution (15 mL / 1.0 mmoL) was added. The reaction was carried out at 30°C for 8 hours. After the reaction was completed by TLC, tetrahydrofuran was removed by reduced pressure concentration, and then 1N hydrochloric acid was used to adjust the pH to 6-7, so that white solids were precipitated. The solids were collected by filtration and dried at 50°C to obtain white powder intermediate a3, light yellow solid, with a yield of 83%. ESI-MS: m / z calcd for C9H 12 BrN2O4S[M+H] + 323.0, found 323.0, 325.0.

[0055] To a solution of intermediate a3 (1.0 eq) and boron trifluoride etherate (1.0 eq) in a mixture of dichloromethane and tetrahydrofuran (5:3, 15 mL / 1.0 mmol) was added tert-butyltrichloroacetimidate (10.0 eq) dropwise. The reaction mixture was stirred at room temperature for 1 h, then concentrated under reduced pressure. The organic phase was collected and purified by column chromatography (petroleum ether: ethyl acetate = 20:1) to give the key intermediate tert-butyl 5-bromo-2-(tert-butoxycarbonyl)amino)thiazole-4-carboxylate (a4, yellowish solid) in 92% yield. ESI-MS: m / z calcd for C 13 H 19 BrN2O4S[M+H] + 379.1, found 379.1, 381.1.

[0056] 2. Synthesis of amide fragment a7’

[0057]

[0058] To a solution of (1R,2S,5R)-6-(benzyloxy)-7-oxo-1,6-diazabicyclo[3.2.1]octane-2- carboxylic acid (a6, 1.0 eq), 2-(7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate (1.5 eq) and triethylamine (1.0 eq) in dichloromethane (15 mL / 1.0 mmol) was added substituted amine a5’ (1.5 eq) after activation for 0.5 h at room temperature. The reaction mixture was stirred at room temperature for 5 h, then concentrated under reduced pressure. The organic phase was collected and purified by column chromatography (petroleum ether: ethyl acetate = 6:1-1:1) to give the amide fragment intermediate a7’ in 65-91% yield. The R in the amide fragment intermediate a7’ was changed according to the structure of target compounds 1-8.

[0059] The following is the synthesis method of target compounds 1-8.

[0060] Example 1: Synthesis of target compound 5

[0061] The synthetic route is as follows:

[0062]

[0063] Step (1): In a three-necked flask, amide intermediate a7 (1.0 eq), intermediate a4 (2.0 eq), cuprous iodide (0.05 eq), triphenylphosphine (0.05 eq) and bis(triphenylphosphine)palladium(II) chloride (0.1 eq) were added to a 1,4-dioxane solution (15 mL / 1.0 mmoL), after argon protection, 100 °C for 6 hours, TLC monitoring until the raw material was completely reacted, concentrated under reduced pressure, extracted with ethyl acetate and water, the organic phase was collected and purified by column chromatography (petroleum ether: ethyl acetate = 3:1) to obtain intermediate a8, light yellow solid, yield 42%. ESI-MS: m / z calcd for C 35 H 39 N5O7S[M+H] + 674.3, found 674.3.

[0064] Step (2): Intermediate a8 (1.0 eq) was dissolved in methanol (15 mL / 1.0 mmoL), then 10% palladium-carbon (2.0 eq) and triethylamine (2.0 eq) were added, and hydrogen was introduced at room temperature and normal pressure for 2 hours. After the reaction was completed, filtration, reduced pressure concentration of the filtrate, column chromatography (dichloromethane:methanol = 50:1) to obtain intermediate a9, white solid, yield 86%. ESI-MS: m / z calcd for C 28 H 37 N5O7S[M+H] + 588.2, found 588.2.

[0065] Step (3): Intermediate a9 (1.0 eq) was dissolved in dichloromethane (15 mL / 1.0 mmoL), then sulfur trioxide pyridine (6.0 eq), triethylamine (10.0 eq) were added respectively, and reacted at 0 °C overnight. After the reaction was completed, reduced pressure concentration, column chromatography (dichloromethane:methanol = 15:1) to obtain intermediate a10, yield 51%, 1 H NMR (400 MHz, DMSO-d6) δ 11.57 (s br, 1H), 10.01 (s, 1H), 9.01 (s br, 1H), 7.61 (d, J = 8.4 Hz, 2H), 7.14 (d, J = 8.4 Hz, 2H), 4.11-4.09 (m, 1H), 4.03 (s, 1H), 3.96 (d, J = 6.8 Hz, 1H), 3.34-3.31 (m, 1H), 3.17 (d, J = 3.2 Hz, 2H), 3.02 (s, 2H), 2.12-2.05 (m, 1H), 1.93-1.90 (m, 1H), 1.80-1.68 (m, 2H), 1.50 (s, 9H), 1.46 (s, 9H) ppm. HRMS m / z: calcd for C28 H 37 N5O 10 S2[M-H] - 666.1909, found 666.1912.

[0066] Step (4): Intermediate a10 was dissolved in dichloromethane (15 mL / 1.0 mmoL), then 15% trifluoroacetic acid in dichloromethane was added, and the reaction was stirred at room temperature for 7 hours. After concentration under reduced pressure, the target compound 5 was obtained by high performance liquid preparative chromatography separation with a yield of 21%. HRMS m / z: calcd for C 19 H 21 N5O8S2[M-H] - 510.0759, found 510.0745.

[0067] Example 2: Synthesis of target compound 1

[0068]

[0069] Step (1): Synthesis of intermediate a12

[0070]

[0071] Vinylamine amide intermediate a11 (1.0 eq), intermediate a4 (2.0 eq), palladium acetate (0.2 eq), tris (o-methylphenyl) phosphine (0.2 eq) and triethylamine (3.0 eq) were added to N,N dimethylformamide (5 mL / 1.0 mmoL), and the reaction was carried out at 90°C for 8 hours under argon protection. After TLC monitoring showed that the raw material was completely reacted, the reaction was concentrated under reduced pressure, extracted with ethyl acetate and water, and the organic phase was collected and purified by column chromatography (petroleum ether: ethyl acetate = 4:1) to obtain intermediate a12, a light yellow solid, with a yield of 32%. ESI-MS: m / z calcd for C 35 H 39 N5O7S[M+H] + 600.3, found 600.3.

[0072] Steps (2)-(4): Refer to the method of steps (2)-(4) of Example 1, and use intermediate a12 to synthesize target compound 1, with a total yield of 11% in three steps. 1H NMR (400 MHz, DMSO-d6) δ 12.06 (s br, 1H), 9.04 (s br, 1H), 6.13 (t, J = 5.6 Hz, 1H), 7.32 (s br, 2H), 3.98 (s, 1H), 3.72 (d, J = 6.8 Hz, 1H), 3.19-3.14 (m, 2H), 3.04-2.99 (m, 3H), 2.93 (d, J = 7.6 Hz, 1H), 2.10-2.04 (m, 1H), 1.86-1.82 (m, 1H), 1.78-1.61 (m, 2H) ppm. HRMS m / z: calcd for C 13 H 17 N5O8S2[M-H] - 434.0440, found 434.0433.

[0073] Example 3: Synthesis of target compound 2

[0074]

[0075] Referring to the method of Example 1, using intermediate (1R,2S,5R)-6- (benzyloxy)-7-oxo-N-(prop-2-ynyl)-1,6-diazabicyclo[3.2.1]octane-2-carboxamide, the target compound 2 was synthesized in a total yield of 9% over four steps. 1 H NMR (400 MHz, DMSO-d6) δ 12.06 (s br, 1H), 9.04 (s br, 1H), 6.13 (t, J = 5.6 Hz, 1H), 7.32 (s br, 2H), 3.98 (s, 1H), 3.72 (d, J = 6.8 Hz, 1H), 3.19-3.14 (m, 2H), 3.04-2.99 (m, 3H), 2.93 (d, J = 7.6 Hz, 1H), 2.10-2.04 (m, 1H), 1.86-1.82 (m, 1H), 1.78-1.61 (m, 2H) ppm. HRMS m / z: calcd for C 14 H 19 N5O8S2[M-H] - 448.0602, found448.0623.

[0076] Example 4: Synthesis of target compound 3

[0077]

[0078] Step (1): Synthesis of intermediate a14

[0079]

[0080] Intermediate a14 was synthesized by the method described in Example 1, steps (2)-(4), using intermediate (1R,2S,5R)-6-(benzyloxy)-7-oxo-N-(4-(4,4,5,5-tetramethyl-1,3,2-dioxobenzol-2-yl)methyl)phenyl)-1,6-diazabicyclo[3.2.1]octane-2-carboxamide (1.0 eq) and intermediate a4 (1.5 eq). The reaction was carried out in 1,4-dioxane (10 mL / 1.0 mmol) with potassium carbonate (2.0 eq) and dichlorobis(triphenylphosphine) palladium (0.2 eq) under argon at 100 °C for 6 h. After the reaction was completed by TLC, the reaction mixture was filtered and the filtrate was concentrated under reduced pressure. The residue was extracted with water and ethyl acetate (x 3), and the combined organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography (dichloromethane:methanol = 35:1) to give intermediate a14.

[0081] Steps (2)-(4): The target compound 3 was synthesized by the method described in Example 1, steps (2)-(4), using intermediate a14. The overall yield was 12% for three steps. HRMS m / z: calcd for C 18 H 18 N5O8S2[M-H] - 496.0601, found 496.0602.

[0082] Example 5: Synthesis of target compound 4

[0083]

[0084] The target compound 4 was synthesized by the method described in Example 4, using intermediate (1R,2S,5R)-6-(benzyloxy)-7-oxo-N-(3-(4,4,5,5-tetramethyl-1,3,2-dioxobenzol-2-yl)methyl)phenyl)-1,6-diazabicyclo[3.2.1]octane-2-carboxamide. The overall yield was 11% for four steps. 1 H NMR (400 MHz, DMSO-d6) δ 12.11 (s br, 1H), 9.72 (s, 1H), 9.05 (s br, 1H), 7.62 (s, 1H), 7.52-7.43 (m, 4H), 7.08 (d, J = 5.6 Hz, 1H), 4.11-4.05 (m, 3H), 3.75-3.58 (m, 5H), 2.11-1.95 (m, 2H), 1.76-1.61 (m, 2H) ppm. C 18 H 18 N5O8S2[M-H] - 496.0601, found 496.0605.

[0085] Example 6: Synthesis of target compound 6

[0086]

[0087] The target compound 6 was synthesized by the method of Example 1 using the intermediate (1R, 2S, 5R)-6-(benzyloxy)-N-(3-ethynylphenyl)-7-oxo-1, 6-diazabicyclo[3.2.1]octane-2-carboxamide, in a total yield of 8% in four steps. HPLC purity 96%, 1 HRMS m / z: calcd for C 19 H 21 N5O8S2[M-H] - 510.0759, found 510.0750.

[0088] Example 7: Synthesis of target compound 7

[0089]

[0090] Step 1: Synthesis of intermediate a17

[0091]

[0092] The intermediate a15 (1.0 eq), cuprous iodide (0.1 eq), triphenylphosphine (0.05 eq), bis(triphenylphosphine)palladium dichloride (0.05 eq) were weighed into a three-necked flask, and dry tetrahydrofuran (10 mL / 1.0 mmol) was added under argon protection. Compound a16 was dissolved in an equal volume of triethylamine in tetrahydrofuran, and slowly added dropwise into the reaction flask. The reaction was stirred at room temperature for half an hour, and the progress of the reaction was monitored by TLC. After the reaction was completed, the solvent was removed by concentration under reduced pressure, and the intermediate a17 was obtained by column chromatography (petroleum ether: ethyl acetate = 3:1) with a yield of 63%. ESI-MS: m / z calcd for C 26 H 31 N3O3Si[M+H] + 462.2, found 462.2.

[0093] Step 2: Synthesis of intermediate a18

[0094] The intermediate a17 (1.0 eq) was dissolved in methanol (15 mL / 1.0 mmol), and 10% aqueous potassium hydroxide solution was added at a volume of 10%. The reaction was stirred at room temperature for 1 hour, and then concentrated under reduced pressure. The intermediate a18 was obtained by column chromatography (petroleum ether: ethyl acetate = 2:1) with a yield of 83%.

[0095] Step 3: Refer to the method of Example 1, Step (1)-(4), the target compound 7 was synthesized from intermediate a18 with a total yield of 8% in four steps. HPLC purity 95%, 1 HRMS m / z: calcd for C 20 H 23 N5O8S2[M-H] - 524.0908, found 524.09018.

[0096] Example 8: Synthesis of target compound 8

[0097]

[0098] Refer to the method of Example 7, the target compound 8 was synthesized from intermediate (1R, 2S, 5R)-6-(benzyloxy)-N-(5-bromothiazol-2-yl)-7-oxo-1,6-diazabicyclo[3.2.1]octane-2-carboxamide (a19) with a total yield of 5% in six steps. HPLC purity 95%, 1 HRMS m / z: calcd for C 16 H 18 N6O8S3[M-H] - 517.0268, found 517.0262.

[0099] The beneficial effects of the present application are demonstrated by the following specific test examples.

[0100] Test Example 1: Inhibitory activity of compounds on MBL and SBL

[0101] 1. Test method

[0102] Test drugs: target compounds 1-8 prepared in Examples 1-8 of the present application, Tazobactam, Avibactam, and Taniborbactam were used as controls.

[0103] The principle of activity test is that MBL and SBL enzymes catalyze the reaction of fluorescent substrate to produce fluorescent groups, and the activity of enzymes can be tested by testing the fluorescence intensity; the activity test is carried out in a black 96-well enzyme plate, and the total reaction system is 60 μL, and the specific operation steps are as follows:

[0104] (1) The test compound is prepared into a 100 mM stock solution with DMSO, and then diluted into a 3.6 mM or 600 μM working solution with MBL activity test buffer (20 mM Tris-HCl pH 7.5, 200 mM NaCl, 0.01% Triton X-100) or SBL activity test buffer (50 mM Phosphate, pH 7.0 or 50 mM HEPES, pH 7.2, 200 mM NaCl, 1 μg / ml BSA). The working solution is diluted three times with the test buffer to obtain 10 different concentrations.

[0105] (2) The MBL (including NDM-1, NDM-5, IMP-1, IMP-4, VIM-1 and VIM-2) and SBL (KPC-2, TEM-1, SHV-12, CTX-M-14, AmpC and OXA-48) to be tested are prepared into enzyme solutions with appropriate concentrations with the activity test buffer.

[0106] (3) The fluorescent substrate FC-5 is prepared into a 30 μM substrate solution with the test buffer for use.

[0107] (4) In the 96-well enzyme-labeled 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 are sequentially added to each well, and incubated at room temperature for 10 minutes.

[0108] (5) After the incubation is completed, 10 μL of the fluorescent substrate solution is added to each well of the 96-well enzyme-labeled plate, and the fluorescence value change is detected continuously for 6 minutes using an enzyme-labeled instrument. The excitation wavelength λ ex of the fluorescence detection is 380 nm, and the emission wavelength λ em is 460 nm.

[0109] (6) A reaction well without the addition of the compound is set as a control for each experiment, and all the determinations contain three parallel experiments.

[0110] According to the change of the fluorescence intensity detected by the enzyme-labeled instrument, the residual activity of the enzyme in each well is calculated, and the calculation formula is: residual activity (%) = (ΔF1) / (ΔF2) x 100, wherein ΔF1 is the fluorescence change value of the well containing the test compound within a certain period of time, and ΔF2 is the fluorescence change value of the control well without the addition of the compound within the same period of time. The processed data is fitted using the Graphpad Prism 5 software to obtain the half-inhibitory activity value (i.e. IC 50 value) of the enzyme.

[0111] 2. Test results

[0112] Table 1. Inhibitory activity IC 50 value of the compound on MBL

[0113]

[0114] Note: +++++: IC 50 <10 nM; +++++: 10 nM < IC 50 <1 μM; +++: 1 μM < IC 50 <100 μM; ++: IC 50 >100 μM.

[0115] Table 2. Inhibitory activity IC of compounds on SBL 50 value

[0116]

[0117] Note: +++++: IC 50 <10 nM; +++++: 10 nM < IC 50 <1 μM; +++: 1 μM < IC 50 <100 μM; ++: IC 50 >100 μM.

[0118] From the above tests, it can be seen that the inhibitory activity of the control drugs tazobactam and avibactam on MBL is poor. However, the compounds of the present application exhibit good and broad-spectrum inhibitory activity on clinically important MBL (including NDM-1, NDM-5, IMP-1, IMP-5, VIM-1 and VIM-2). Among them, the inhibitory activity of compounds 3, 4, 5, 6, 7 and 8 on NDM-1, NDM-5, VIM-1 and / or VIM-2 reaches the nanomolar level; the inhibitory activity of compounds 3 and 6 on IMP-1 and / or IMP-5 reaches the nanomolar level. Compared with the control drugs tazobactam and avibactam, the compounds of the present application have better inhibitory activity on MBL. The inhibitory activity of multiple compounds (such as 3, 4, 5, 6, 7 and 8) of the present application on MBL is comparable to that of the tienamstat which is in the clinical evaluation stage.

[0119] At the same time, the compounds of the present application also exhibit good and broad-spectrum inhibitory activity on clinically important SBL (including KPC-2, TEM-1, SHV-12, CTX-M-14, AmpC and OXA-48). The inhibitory activity of all compounds of the present application on KPC-2, TEM-1, CTX-M-14, SHV-12, AmpC and OXA-48 is at the nanomolar level. Among them, the inhibitory activity of some compounds (such as 2, 5 and 7) on KPC-2, SHV-12, AmpC and OXA-48 is better than that of the control drugs tazobactam and tienamstat, and is comparable to that of avibactam.

[0120] The above experimental results show that the compound provided by the application has good and broad-spectrum inhibitory activity on common MBL and SBL in clinic, and can be used as an inhibitor of MBL and / or SBL, and can be used for preparing antibacterial drugs, in particular, drugs for resisting drug-resistant bacteria.

[0121] In conclusion, the application provides a class of thiazole amine-diazabicyclooctanone conjugate derivatives and uses thereof. The compound provided by the application has good and broad-spectrum inhibitory activity on MBL and SBL, and can be used for preparing an inhibitor of MBL and / or SBL. The compound of the application has great potential in preparing an inhibitor of MBL and / or SBL and an antibacterial drug, in particular, a drug for resisting drug-resistant bacteria.

Claims

1. A compound or a pharmaceutically acceptable salt thereof, characterized in that: The structure of the compound is shown in Formula III: Wherein, x1 is selected from 1, 2; y1 is selected from 0, 1; m is selected from 0, 1, 2, 3; R1 are each independently selected from hydrogen, C 1-8 Alkyl, C 1-8 Alkoxy, halogen, hydroxy, amino, carboxyl, cyano.

2. A compound or a pharmaceutically acceptable salt thereof, characterized in that: The structure of the compound is shown in Formula IV: Wherein, x2 is selected from 1, 2; y2 is selected from 0, 1; R1 is selected from hydrogen, C 1-8 Alkyl, C 1-8 Alkoxy, halogen, hydroxy, amino, carboxyl, cyano; Z is selected from O, S or CH2.

3. A compound or a pharmaceutically acceptable salt thereof, characterized in that: The compound is selected from:

4. An antibacterial drug, characterized in that: The invention is prepared by taking the compound according to any one of claims 1 to 3 or a pharmaceutically acceptable salt thereof as an active ingredient and adding pharmaceutically acceptable excipients.

5. Use of the compound according to any one of claims 1 to 3 or a pharmaceutically acceptable salt thereof in the preparation of a β-lactamase inhibitor.

6. The use according to claim 5, characterized in that: The β-lactamase inhibitor is a metallo-β-lactamase inhibitor, a serine-β-lactamase inhibitor, or a metallo-β-lactamase and serine-β-lactamase dual inhibitor.

7. The use according to claim 5 or 6, characterized in that: The β-lactamase inhibitor is an antibacterial drug.

8. The use according to claim 7, characterized in that: The antibacterial drug is a drug for resisting drug-resistant bacteria.

9. The use according to claim 8, characterized in that: The drug-resistant bacteria are bacteria resistant to β-lactam antibiotics.

10. A combined drug with antibacterial efficacy, characterized in that: It contains the compound or pharmaceutically acceptable salt thereof and a β-lactam antibiotic according to any one of claims 1 to 3, and a pharmaceutically acceptable carrier, in unit preparations of the same or different specifications for simultaneous or separate administration.

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