WO2024007883A1 - Class of thiazolamine-diazabicyclooctanone conjugated derivatives and use thereof - Google Patents

Abstract

The present invention belongs to the field of medicines and provides a class of thiazolamine-diazabicyclooctanone conjugated derivatives and use thereof. The compound provided by the present invention has good and broad-spectrum inhibition activity on metal β-lactamase (MBL) and serine β-lactamase (SBL) and can be used for preparing an inhibitor for MBL and/or SBL. The compound of the present invention has very great potential for preparing the inhibitor for MBL and/or SBL and antibacterial drugs (especially drugs used to combat drug-resistant bacteria).

Description

A class of thiazolamine-diazabicyclooctanone conjugated derivatives and their uses

This application requires the priority of the Chinese patent application submitted to the China Patent Office on July 5, 2022, with the application number 202210783672.9 and the invention title "A class of thiazolamine-diazabicyclooctanone conjugated derivatives and their uses" , the entire contents of which are incorporated herein by reference.

Technical field

The invention belongs to the field of medicine, and specifically relates to a class of thiazolamine-diazabicyclooctanone conjugated derivatives and their uses.

Background technique

β-lactam antibiotics are currently the most widely used antibiotics in clinical practice, such as penicillins, cephalosporins, carbapenems, monocyclics, etc. Among them, carbapenem antibiotics are a class of β-lactam antibiotics with the broadest antibacterial spectrum and the strongest bactericidal ability. They are one of the most important drugs for the clinical treatment of serious bacterial infections and were once known as the "last resort" against bacterial infections. A line of defense.” However, at present, resistance to β-lactam antibiotics is very common, and the resistance situation is very serious. In the list of drug-resistant bacteria published by the World Health Organization in 2017, carbapenem-resistant Acinetobacter baumannii, Pseudomonas aeruginosa and Enterobacteriaceae are listed as the most dangerous superbugs, and effective treatments are still lacking. Drugs have attracted widespread attention and vigilance around the world. Studies have found that the main mechanism leading to resistance to β-lactam antibiotics, including carbapenems, is that the pathogenic bacteria themselves produce β-lactamase, which hydrolyzes the core efficacy skeleton of the β-lactam ring, thereby causing it to lose its antibacterial activity. .

According to the Ambler classification, β-lactamases can be divided into 4 types: A, B, C, and D. Among them, the active center of types A, C, and D is serine, and they are collectively called serine β-lactamases (SBL). ; The active center of class B contains at least one Zn ²⁺ , so class B β-lactamase is also called metalloβ-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 practice. The β-lactamase inhibitors currently used clinically mainly include clavulanic acid, sulbactam, tazobactam and the newly launched Avibactam. Clavulanic acid, sulbactam, and tazobactam are all irreversible inhibitors that only inhibit most class A beta-lactamases and have no inhibitory effect on class B, C, and D beta-lactamases. Avibactam is a reversible inhibitor that inhibits class A, class C and some class D beta-lactamases. However, its inhibitory effect on class B beta-lactamases is not good.

Avibataan

Due to the exchange and transfer of genetic material between genes expressing drug-resistant enzymes between the same or different pathogenic bacteria, drug-resistant bacteria co-expressed by MBL and SBL continue to appear and spread, posing a huge threat to human life and health. In the face of the increasing number of multi-drug-resistant, extensively-drug-resistant (XDR) and even pan-drug-resistant (PDR) “superbugs”, existing β-lactamase inhibitors are basically Unable to meet clinical needs, it is urgent to develop inhibitors with dual inhibitory effects on MBL and SBL.

Contents of the invention

The object of the present invention is to provide a class of thiazolamine-diazabicyclooctanone conjugated derivatives and their preparation methods and uses.

The present invention provides a compound, its conformational isomer, its optical isomer or its pharmaceutically acceptable salt. The structure of the compound is shown in Formula I:

Among them, L is a linker or none.

Further, the L is selected from C _1-8 alkylene,

X is selected from none and C _1-8 alkylene; Y is selected from none and C _1-8 alkylene;

Ring A is selected from the group consisting of 3- to 6-membered saturated cycloalkyl, 3- to 6-membered saturated heterocyclic, 5- to 6-membered aryl, 5- to 6-membered heteroaryl, fused cycloalkyl, and heterofused cyclic;

m is selected from 0, 1, 2, 3;

R ₁ is each independently selected from hydrogen, C _1-8 alkyl, C _1-8 alkoxy, halogen, hydroxyl, amino, carboxyl, cyano.

Further, the structure of the compound is shown in Formula II:

Among them, n is selected from 0, 1, 2, 3, 4, 5.

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

Among them, x1 is selected from 0, 1, 2, 3, 4, 5;

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

m is selected from 0, 1, 2, 3;

R ₁ is each independently selected from hydrogen, C _1-8 alkyl, C _1-8 alkoxy, halogen, hydroxyl, amino, carboxyl, cyano.

Further, the structure of the compound is shown in Formula IV:

Among them, x2 is selected from 0, 1, 2, 3, 4, 5;

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

R ₁ is selected from hydrogen, C _1-8 alkyl, C _1-8 alkoxy, halogen, hydroxyl, amino, carboxyl, and cyano;

Z is selected from O, S or _CH2 .

Further, the compound is selected from:

The invention provides an antibacterial drug, which is prepared by using the above compound, its conformational isomer, its optical isomer or its pharmaceutically acceptable salt as active ingredients, and adding pharmaceutically acceptable auxiliary materials. .

The present invention also provides the use of the above-mentioned compound, its conformational isomer, its optical isomer or its pharmaceutically acceptable salt in the preparation of β-lactamase inhibitors.

Further, the β-lactamase inhibitor is a metalloβ-lactamase inhibitor, a serine β-lactamase inhibitor, a metalloβ-lactamase and a serine β-lactamase dual inhibitor.

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

Further, the antibacterial drugs are drugs against drug-resistant bacteria, and the drug-resistant bacteria are preferably bacteria resistant to β-lactam antibiotics.

The present invention also provides a combination drug with antibacterial efficacy, which contains the above-mentioned compound, its conformational isomer, its optical isomer or its pharmaceutically active form in unit preparations of the same or different specifications for simultaneous or separate administration. Acceptable salts and beta-lactam antibiotics, and pharmaceutically acceptable carriers.

Definitions of terms used in the present invention: Unless otherwise stated, the initial definition provided for a group or term in this article applies to the group or term in the entire specification; for terms that are not specifically defined in this article, it should be based on the disclosure content and context. , giving the meanings that those skilled in the art can give them.

The minimum and maximum content of carbon atoms in a hydrocarbon group is indicated by a prefix, for example, the prefix C _{a to b} alkyl means any alkyl group containing "a" to "b" carbon atoms. For example, C _1-8 alkyl refers to a linear or branched alkyl group containing 1 to 8 carbon atoms.

"Aryl" refers to an all-carbon monocyclic group having a conjugated pi electron system, such as phenyl. The aryl group does not contain heteroatoms such as nitrogen, oxygen, or sulfur, and the point of attachment to the parent must be on a carbon atom in the ring with a conjugated pi electron system.

"Heteroaryl" refers to a heteroaromatic group containing one to more heteroatoms. Heteroatoms referred to here include oxygen, sulfur and nitrogen. For example, furyl, thienyl, pyridyl, pyrazolyl, pyrrolyl, N-alkylpyrrolyl, pyrimidinyl, pyrazinyl, imidazolyl, tetrazolyl, etc.

"Fused cycloalkyl" refers to a polycyclic cycloalkyl group in which two rings share two adjacent carbon atoms.

"Heterofused cyclyl" refers to a polycyclic heterocyclyl, and two of the polycyclic heterocyclyl groups share two adjacent carbon atoms or heteroatoms.

Halogen is fluorine, chlorine, bromine or iodine.

The connector is Linker.

The compound provided by the invention has good and broad-spectrum inhibitory activity against metallo-β-lactamase (MBL) and serine β-lactamase (SBL), and can be used to prepare inhibitors of MBL and/or SBL. The compounds of the present invention are useful in preparing inhibitors of MBL and/or SBL and antibacterial drugs (especially drugs against drug-resistant bacteria). Has great potential.

Obviously, according to the above content of the present invention, according to the common technical knowledge and common means in the field, without departing from the above basic technical idea of the present invention, various other forms of modifications, replacements or changes can also be made.

The above contents of the present invention will be further described in detail below through specific implementation methods in the form of examples. However, this should not be understood to mean that the scope of the above subject matter of the present invention is limited to the following examples. All technologies implemented based on the above contents of the present invention belong to the scope of the present invention.

Detailed ways

The raw materials and equipment used in the present invention are all known products and are obtained by purchasing commercially available products.

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

1.Synthesis of intermediate a4

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

Dissolve intermediate a2 (1.0eq) in tetrahydrofuran solution (15mL/1.0mmoL), then add 10% potassium hydroxide aqueous solution (15mL/1.0mmoL), and react at 30°C for 8 hours. TLC monitors that the reaction is complete and then concentrates under reduced pressure to remove tetrahydrofuran. Then adjust the pH to 6-7 with 1N hydrochloric acid and a white solid will precipitate. Collect the solid by filtration and dry it at 50°C to obtain white powder intermediate a3, a light yellow solid, yield 83 %. ESI-MS: m/z calcd for C ₉ H ₁₂ BrN ₂ O ₄ S[M+H] ⁺ 323.0, found 323.0, 325.0.

After adding intermediate a3 (1.0eq) and boron trifluoride ether (1.0eq) to the mixed solution of dichloromethane and tetrahydrofuran (5:3, 15mL/1.0mmoL), slowly drop in tert-butyltrichloroacetyl Amino ester (10.0eq), reacted at room temperature for 1 hour, concentrated under reduced pressure, extracted with saturated sodium bicarbonate solution, collected the organic phase, and purified by column chromatography (petroleum ether: ethyl acetate = 20:1) to obtain key intermediate 5 -Bromo-2-(tert-butoxycarbonyl)amino)thiazole-4-carboxylic acid tert-butyl ester (a4, light yellow solid), yield 92%. ESI-MS: m/z calcd for C ₁₃ H ₁₉ BrN ₂ O ₄ S[M+H] ⁺ 379.1, found 379.1, 381.1.

2. Synthesis of amide fragment a7'

(1R, 2S, 5R)-6-(benzyloxy)-7-oxy-1,6-diazabicyclo[3.2.1]octane-2-carboxylic acid (a6, 1.0eq), 2-(7-Azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (1.5eq) and triethylamine (1.0eq) were added to dichloromethane ( 15mL/1.0mmoL) was dissolved and activated for 0.5 hours under stirring at room temperature. After adding substituted amine a5' (1.5eq), the reaction was continued with stirring at room temperature for 5 hours, and then concentrated under reduced pressure. Column chromatography (petroleum ether: ethyl acetate) =6:1-1:1) to obtain the amide fragment intermediate a7', with a yield of 65%-91%. The R in the amide fragment intermediate a7' is changed accordingly according to the structure of the target compound 1-8.

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

Example 1: Synthesis of target compound 5

The synthesis route is:

Step (1): In a three-necked flask, prepare the amide intermediate a7 (1.0eq) of p-alkylenyl aniline, intermediate a4 (2.0eq), copper iodide (0.05eq), triphenylphosphine (0.05eq) and bis(triphenylphosphine)palladium dichloride (0.1eq) were added to the 1,4-dioxane solution (15mL/1.0mmoL), and then heated to 100°C for 6 hours under argon protection. The raw materials were monitored by TLC. After the reaction is complete, concentrate under reduced pressure and extract with ethyl acetate and water. The organic phase is collected and purified by column chromatography (petroleum ether: ethyl acetate = 3:1) to obtain intermediate a8, a light yellow solid, with a yield of 42%. . ESI-MS: m/z calcd for C ₃₅ H ₃₉ N ₅ O ₇ S[M+H] ⁺ 674.3, found 674.3.

Step (2): After dissolving intermediate a8 (1.0eq) with methanol (15mL/1.0mmoL), add 10% palladium on carbon (2.0eq) and triethylamine (2.0eq), and then add hydrogen at room temperature and pressure. The reaction was carried out for 2 hours, filtered after the reaction was complete, and the filtrate was concentrated under reduced pressure. After column chromatography (dichloromethane:methanol=50:1), intermediate a9 was obtained as a white solid, with a yield of 86%. ESI-MS: m/z calcd for C ₂₈ H ₃₇ N ₅ O ₇ S[M+H] ⁺ 588.2, found588.2.

Step (3): After dissolving intermediate a9 (1.0eq) with dichloromethane (15mL/1.0mmoL), add sulfur trioxide pyridine (6.0eq) and triethylamine (10.0eq) respectively, and react at 0°C overnight. , after the reaction was completed, it was concentrated under reduced pressure and purified by column chromatography (dichloromethane: methanol = 15:1) to obtain intermediate a10, yield 51%, ¹ H NMR (400MHz, DMSO-d ₆ ) δ 11.57 (s br,1H),10.01(s,1H),9.01(s br,1H),7.61(d,J＝8.4Hz,2H),7.14(d,J＝8.4Hz,2H),4.11-4.09(m, 1H),4.03(s,1H),3.96(d,J＝6.8Hz,1H),3.34-3.31(m,1H),3.17(d,J＝3.2Hz,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 C ₂₈ H ₃₇ N ₅ O ₁₀ S ₂ [MH] ^- 666.1909,found 666.1912.

Step (4): Dissolve intermediate a10 with dichloromethane (15mL/1.0mmoL), add an equal volume of 15% trifluoroacetic acid in dichloromethane, stir and react at room temperature for 7 hours, concentrate under reduced pressure, and react with high performance liquid phase The target compound 5 was separated by preparative chromatography with a yield of 21%. HRMS m/z:calcd for C ₁₉ H ₂₁ N ₅ O ₈ S ₂ [MH] ^- 510.0759,found 510.0745.

Example 2: Synthesis of target compound 1

Step (1): Synthesis of intermediate a12

Add vinylamine amide intermediate a11 (1.0eq), intermediate a4 (2.0eq), palladium acetate (0.2eq), tris(o-methylphenyl)phosphorus (0.2eq) and triethylamine (3.0eq) to N,N dimethylformamide (5mL/1.0mmoL), and then reacted at 90°C for 8 hours under argon protection. After the raw materials were completely reacted and monitored by TLC, they were concentrated under reduced pressure and extracted with ethyl acetate and water. The organic phase was collected. After purification by column chromatography (petroleum ether: ethyl acetate = 4:1), intermediate a12 was obtained as a light yellow solid, with a yield of 32%. ESI-MS: m/z calcd for C ₃₅ H ₃₉ N ₅ O ₇ S[M+H] ⁺ 600.3, found 600.3.

Steps (2)-(4): Referring to the method of steps (2)-(4) in Example 1, the target compound 1 was synthesized using intermediate a12. The total yield of the three steps was 11%. ¹ H NMR (400MHz, DMSO-d ₆ ) δ 12.06 (s br, 1H), 9.04 (s br, 1H), 6.13 (t, J = 5.6Hz, 1H), 7.32 (s br, 2H), 3.98 (s,1H),3.72(d,J＝6.8Hz,1H),3.19-3.14(m,2H),3.04-2.99(m,3H),2.93(d,J＝7.6Hz,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 N 5 O 8} S ₂ [MH] ^- 434.0440, found 434.0433.

Example 3: Synthesis of target compound 2

Referring to the method of Example 1, the intermediate (1R, 2S, 5R)-6-(benzyloxy)-7-oxy-N-(prop-2-ynyl)-1,6-diazabi Cycl[3.2.1]octane-2-carboxamide was used to synthesize target compound 2, with a total yield of 9% in four steps. ¹ H NMR (400MHz, DMSO-d ₆ ) δ 12.08 (s br, 1H), 9.03 (s br, 1H), 8.15 (t, J = 5.6Hz, 1H), 7.32 (s br, 2H), 4.02 -3.91(m,2H),3.74-3.52(m,2H),3.19-3.13(m,2H),2.52-2.41(m,2H),2.10-1.98(m,4H),1.80-1.66(m, 2H)ppm.HRMS m/z:calcd for C ₁₄ H ₁₉ N ₅ O ₈ S ₂ [MH] ^- 448.0602, found 448.0623.

Example 4: Synthesis of target compound 3

Step (1): Synthesis of intermediate a14

(1R, 2S, 5R)-6-(benzyloxy)-7-oxo-N-(4-(4,4,5,5-tetramethyl-1,3,2-dioxobenzaldehyde -2-yl)methyl)phenyl)-1,6-diazabicyclo[3.2.1]octane-2-carboxamide (1.0eq) and intermediate a4 (1.5eq) are dissolved in 1,4 - Dioxane (10mL/1.0mmoL), then add potassium carbonate (2.0eq), bistriphenylphosphine palladium dichloride (0.2eq), under argon protection, raise the temperature to 100°C and react for 6 hours. After TLC monitors that the reaction is complete, filter the reaction solution to collect the filtrate. After the solvent is evaporated under reduced pressure, extract with water and ethyl acetate (×3). Combine the organic layers, wash with saturated brine, dry over anhydrous sodium sulfate, and concentrate under reduced pressure. After purification by column chromatography (dichloromethane: methanol = 35:1), intermediate a14 was obtained.

Steps (2)-(4): Referring to the method of steps (2)-(4) in Example 1, the target compound 3 was synthesized using intermediate a14. The total yield of the three steps was 12%. HRMS m/z:calcd for C ₁₈ H ₁₈ N ₅ O ₈ S ₂ [MH] ^- 496.0601, found 496.0602.

Example 5: Synthesis of target compound 4

Referring to the method of Example 4, the intermediate (1R, 2S, 5R)-6-(benzyloxy)-7-oxy-N- (3-(4,4,5,5-Tetramethyl-1,3,2-dioxobenzaldehyde-2-yl)methyl)phenyl)-1,6-diazabicyclo[3.2. 1] Octane-2-carboxamide, the target compound 4 was synthesized, and the total yield in four steps was 11%. ¹ H NMR (400MHz, DMSO-d ₆ ) δ12.11(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＝5.6Hz,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 N ₅ O ₈ S ₂ [MH] ^- 496.0601, found 496.0605.

Example 6: Synthesis of target compound 6

Referring to the method of Example 1, the intermediate (1R, 2S, 5R)-6-(benzyloxy)-N-(3-ethynylphenyl)-7-oxy-1,6-diaza Bicyclo[3.2.1]octane-2-carboxamide intermediate was used to synthesize target compound 6, with a total yield of 8% in four steps. HPLC purity 96%, ¹ HRMS m/z:calcd for C ₁₉ H ₂₁ N ₅ O ₈ S ₂ [MH] ^- 510.0759, found 510.0750.

Example 7: Synthesis of target compound 7

Step 1: Synthesis of intermediate a17

Weigh the intermediate a15 (1.0eq), copper iodide (0.1eq), triphenylphosphine (0.05eq), and bis(triphenylphosphine)palladium dichloride (0.05eq) into a three-necked flask respectively. , add dry tetrahydrofuran (10mL/1.0mmoL) after argon protection, then dissolve compound a16 into an equal amount of triethylamine as the tetrahydrofuran solution, slowly drip it into the reaction bottle, stir and react at room temperature for half an hour, and monitor the reaction progress with TLC. After the reaction was completed, the solvent was concentrated under reduced pressure, and then purified by column chromatography (petroleum ether: ethyl acetate = 3:1) to obtain intermediate a17, with a yield of 63%. ESI-MS: m/z calcd for C ₂₆ H ₃₁ N ₃ O ₃ Si[M+H] ⁺ 462.2, found 462.2.

Step 2: Synthesis of intermediate a18

After adding intermediate a17 (1.0eq) to methanol (15mL/1.0mmoL) to dissolve, add 10% potassium hydroxide aqueous solution with 10% solvent volume, stir and react at room temperature for 1 hour, concentrate under reduced pressure and perform column chromatography ( stone After separation and purification (oil ether: ethyl acetate = 2:1), intermediate a18 was obtained with a yield of 83%.

Step 3: Referring to the method of steps (1) to (4) of Example 1, the target compound 7 was synthesized using intermediate a18. The total yield of the four steps was 8%. HPLC purity 95%, ¹ HRMS m/z:calcd for C ₂₀ H ₂₃ N ₅ O ₈ S ₂ [MH] ^- 524.0908, found 524.09018.

Example 8: Synthesis of target compound 8

Referring to the method of Example 7, the intermediate (1R, 2S, 5R)-6-(benzyloxy)-N-(5-bromothiazol-2-yl)-7-oxo-1,6-diazo The target compound 8 was synthesized from heterobicyclo[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 N ₆ O ₈ S ₃ [MH] ^- 517.0268, found 517.0262.

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

Test Example 1: Inhibitory activity of compounds against MBL and SBL

1. Test method

Test drugs: target compounds 1-8 prepared in Examples 1-8 of the present invention, with Tazobactam, Avibactam, and Taniborbactam as controls.

The principle of the activity test is that MBL and SBL enzymes catalyze the reaction of fluorescent substrates to produce fluorescent groups. Testing the fluorescence intensity can reflect the activity of the enzyme. The activity test is carried out in a black 96-well enzyme plate. The total reaction system is 60 μL. The specific operation steps are as follows :

(1) Use DMSO to prepare the solid compound to be tested into a mother solution with a concentration of 100mM, and use MBL activity test buffer (20mM Tris-HCl pH 7.5, 200mM NaCl, 0.01% Triton X-100) or SBL activity test buffer (50mM Phosphate, pH 7.0 or 50mM HEPES, pH 7.2, 200mM NaCl, 1μg/ml BSA), dilute the mother solution into a 3.6mM or 600μM working solution, and then dilute the working solution three times with the test buffer to obtain 10 working solutions with different concentrations. liquid.

(2) Combine the MBL to be tested (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), use activity test buffer to prepare an enzyme solution of appropriate concentration.

(3) Use test buffer to prepare the fluorescent substrate FC-5 into a 30 μM substrate solution for later use.

(4) In a 96-well microplate, add 10 μL of the compound working solution obtained in step (1), 30 μL test buffer, and 10 μL enzyme solution to each well, and incubate at room temperature for 10 minutes.

(5) After the incubation is completed, add 10 μL of fluorescent substrate solution to each well of the 96-well microplate, and use a microplate reader to continuously detect the changes in fluorescence value within 6 minutes. The excitation wavelength λ _ex of the fluorescence detection is 380 nm, and the emission wavelength λ _em is 460nm.

(6) In each experiment, a reaction well without compound was set up as a control, and all measurements included three sets of parallel experiments.

According to the change in fluorescence intensity measured by the microplate reader, the residual activity of the enzyme in each well is calculated. The calculation is published as: Residual activity (%) = (ΔF1)/(ΔF2) × 100, where 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 compound within the same time period. The processed data were fitted using Graphpad Prism 5 software, and the half inhibitory activity value (i.e. IC ₅₀ value) of the enzyme was calculated.

2. Test results

Table 1. IC ₅₀ value of compound’s inhibitory activity against MBL

Note: +++++: IC ₅₀ <10nM; ++++: 10nM<IC ₅₀ <1μM; +++: 1μM<IC ₅₀ <100μM; ++:
_IC50 >100μM.

Table 2. Inhibitory activity IC ₅₀ value of compounds against SBL

Note: +++++: IC ₅₀ <10nM; ++++: 10nM<IC ₅₀ <1μM; +++: 1μM<IC ₅₀ <100μM; ++:
_IC50 >100μM.

It can be seen from the above experiments that the control drugs tazobactam and avibactam have poor inhibitory activity on MBL. However, the compounds of the present invention have no effect on clinically important MBL (including NDM-1, NDM-5, IMP-1, IMP-5, VIM-1 and VIM-2) showed good and broad-spectrum inhibitory activity. Among them, compounds 3, 4, 5, 6, 7 and 8 have nanomolar inhibitory activity against NDM-1, NDM-5, VIM-1 and/or VIM-2; compounds 3 and 6 have inhibitory activity against IMP-1 and/or VIM-2. Or the inhibitory activity of IMP-5 reaches the nanomolar level. Compared with the control drugs tazobactam and avibactam, the compounds of the present invention have better inhibitory activity against MBL. The inhibitory activity of multiple compounds of the present invention (such as 3, 4, 5, 6, 7 and 8) on MBL is equivalent to that of taniborbactam, which is in the clinical evaluation stage.

At the same time, the compounds of the present invention also show good and broad-spectrum inhibitory activity against clinically important SBLs (including KPC-2, TEM-1, SHV-12, CTX-M-14, AmpC and OXA-48). The inhibitory activities of all compounds of the present invention on KPC-2, TEM-1, CTX-M-14, SHV-12, AmpC and OXA-48 are at the nanomolar level. The inhibitory activity of some compounds (such as 2, 5 and 7) against KPC-2, SHV-12, AmpC and OXA-48 is better than that of the control drugs tazobactam and taniborbactam, and is comparable to that of avibactam. Quite active.

The above experimental results show that the compounds provided by the present invention have good and broad-spectrum inhibitory activity against MBL and SBL, which are common in clinical practice, and can be used as inhibitors of MBL and/or SBL for the preparation of antibacterial agents, especially those against drug-resistant bacteria. drug.

In summary, the present invention provides a type of thiazolamine-diazabicyclooctanone conjugated derivatives and their uses. The compound provided by the invention has good and broad-spectrum inhibitory activity against MBL and SBL, and can be used to prepare inhibitors of MBL and/or SBL. The compounds of the present invention have great potential in preparing inhibitors of MBL and/or SBL and antibacterial drugs (especially drugs against drug-resistant bacteria).

Claims (12)

A compound, its conformational isomer, its optical isomer or its pharmaceutically acceptable salt, characterized in that: the structure of the compound is as shown in formula I:
Among them, L is a linker or none. The compound according to claim 1, its conformational isomer, its optical isomer or its pharmaceutically acceptable salt, characterized in that: said L is selected from C _1-8 alkylene, X is selected from none and C _1-8 alkylene; Y is selected from none and C _1-8 alkylene; Ring A is selected from the group consisting of 3- to 6-membered saturated cycloalkyl, 3- to 6-membered saturated heterocyclic, 5- to 6-membered aryl, 5- to 6-membered heteroaryl, fused cycloalkyl, and heterofused cyclic; m is selected from 0, 1, 2, 3; R ₁ is each independently selected from hydrogen, C _1-8 alkyl, C _1-8 alkoxy, halogen, hydroxyl, amino, carboxyl, cyano. The compound according to claim 2, its conformational isomer, its optical isomer or its pharmaceutically acceptable salt, characterized in that: the structure of the compound is shown in formula II:
Among them, n is selected from 0, 1, 2, 3, 4, 5. The compound according to claim 2, its conformational isomer, its optical isomer or its pharmaceutically acceptable salt, characterized in that: the structure of the compound is shown in formula III:
Among them, x1 is selected from 0, 1, 2, 3, 4, 5; y1 is selected from 0, 1, 2, 3, 4, 5; m is selected from 0, 1, 2, 3; R ₁ is each independently selected from hydrogen, C _1-8 alkyl, C _1-8 alkoxy, halogen, hydroxyl, amino, carboxyl, cyano. The compound according to claim 2, its conformational isomer, its optical isomer or its pharmaceutically acceptable salt, characterized in that: the structure of the compound is as shown in formula IV:
Among them, x2 is selected from 0, 1, 2, 3, 4, 5; y2 is selected from 0, 1, 2, 3, 4, 5; R ₁ is selected from hydrogen, C _1-8 alkyl, C _1-8 alkoxy, halogen, hydroxyl, amino, carboxyl, and cyano; Z is selected from O, S or _CH2 . The compound according to claim 1, its conformational isomer, its optical isomer or its pharmaceutically acceptable salt, characterized in that: the compound is selected from:

An antibacterial drug, characterized in that: it uses the compound described in any one of claims 1 to 6, its conformational isomer, its optical isomer or its pharmaceutically acceptable salt as an active ingredient, plus pharmaceutical prepared with acceptable excipients. Use of the compound according to any one of claims 1 to 6, its conformational isomer, its optical isomer or its pharmaceutically acceptable salt in the preparation of β-lactamase inhibitors. The use according to claim 8, characterized in that: the β-lactamase inhibitor is a metalloβ-lactamase inhibitor, a serine β-lactamase inhibitor, a metalloβ-lactamase and serine β -Dual lactamase inhibitor. The use according to claim 8 or 9, characterized in that the β-lactamase inhibitor is an antibacterial drug. The use according to claim 10, characterized in that: the antibacterial drugs are drugs against drug-resistant bacteria, and the drug-resistant bacteria are preferably bacteria resistant to β-lactam antibiotics. A combination drug with antibacterial efficacy, characterized in that it contains the compound of any one of claims 1 to 6, its conformational isomers, and its conformational isomers in unit preparations of the same or different specifications for simultaneous or separate administration. Optical isomers or pharmaceutically acceptable salts thereof and β-lactam antibiotics, and pharmaceutically acceptable carriers.