A class of compounds with antibacterial uses

Abstract

The present invention belongs to the field of pharmaceutical technology and relates to a class of compounds with antibacterial uses, specifically N-allyl-substituted indoline-2-one-3-hydrazine dithiocarboxylate derivatives. The compounds are shown as formula (I): wherein, R1 represents a C1-C4 alkyl group, and R4, R5, R6 and R7 represent a halogen, a C1-C4 alkyl group, a C1-C4 alkoxy group or a trifluoromethoxy group. The compounds involved in the present invention have the activity of inhibiting New Delhi metallo-β-lactamase 1 (NDM-1). When combined with β-lactam antibiotics such as meropenem, they can restore the sensitivity of superbugs to the corresponding antibiotics and can be used as antibacterial drugs.

Description

Technical Field

[0001] The present invention belongs to the field of pharmaceutical technology, and relates to a class of compounds with antibacterial uses, specifically N-allyl-substituted indoline-2-one-3-hydrazine dithiocarboxylate derivatives, a preparation method thereof, and their application in antibacterial drugs. Background Art

[0002] The discovery and development of antibiotics are the greatest breakthroughs in the 20th century. In recent years, due to reasons such as the unreasonable use of antibiotics, bacteria resistant to antibiotics have emerged continuously, increasing the treatment difficulty and cost of bacterial infectious diseases, thus posing a serious threat to human health. As one of the main antibiotics, β-lactam antibiotics (penicillins, carbapenems, cephalosporins, and monobactams) have the characteristics of broad spectrum, high efficiency, and low toxicity, and are therefore one of the most important and commonly used antibiotics in clinical practice. However, with the emergence and spread of β-lactamases (BLs), the clinical application of this class of drugs has faced unprecedented challenges. β-lactamase (MBL) degrades β-lactam antibiotics by catalytic hydrolysis to cleave the β-lactam bond, thereby generating drug resistance.

[0003] According to the homology of amino acid sequences, β-lactamases can be classified into four types: A, B, C, and D. Types A, C, and D are serine-β-lactamases, which act by hydrolysis at the active site serine, and mainly appear in clinically relevant TEM, SHV, CTX-M, and KPC type variants (type A), AmpC and plasmid-encoded CMY type cephalosporinases (type C), and OXA type enzymes (oxacillinases, type D). Class B β-lactamases rely on one or two zinc ions to hydrolyze the β-lactam ring and are therefore called metallo-β-lactamases (MBLs). NDM-1 (New Delhi metallo-β-lactamase-1), a member of the metallo-β-lactamase family, got its name because the first superbug infection carrying metallo-β-lactamase occurred in New Delhi, India. New Delhi metallo-β-lactamase 1 can degrade almost all β-lactam antibiotics (such as penicillins, cephalosporins, and carbapenems), so the bacterial drug resistance problem it brings is more serious. Bacteria carrying NDM-1 mainly include Escherichia coli, Klebsiella pneumoniae, Acinetobacter baumannii, Enterobacter cloacae, Proteus spp., etc., often causing infections in parts such as the urinary tract and lungs.

[0004] By developing metal-β-lactamase inhibitors, the degradation of β-lactam antibiotics by superbugs can be effectively inhibited. When combined with existing β-lactam antibiotics, the sensitivity of superbugs to β-lactam antibiotics can be restored. Although more than 500 metal-β-lactamase inhibitors have been reported in the literature, there are no marketed drugs, and few compounds have entered clinical trials. Therefore, there is an urgent need to develop new structural types of NDM-1 inhibitors to overcome β-lactam antibiotic resistance and meet clinical needs. Summary of the Invention

[0005] The compounds involved in this patent belong to N-allyl-substituted indoline-2-one-3-hydrazine dithiocarboxylate derivatives, which have obvious activity in inhibiting New Delhi metallo-β-lactamase 1 (NDM-1). When combined with meropenem, they can restore the sensitivity of Escherichia coli ATCC BAA-2469 producing NDM-1 enzyme to meropenem, and have the value of further development. The technical problem solved by this invention is to overcome the defect of insufficient anti-drug-resistant bacteria drugs in the prior art, and provide N-allyl-substituted indoline-2-one-3-hydrazine dithiocarboxylate derivatives, preparation methods and their antibacterial uses.

[0006] To solve the technical problems of this invention, the following technical solutions are provided by this invention:

[0007] In the first aspect, a compound represented by formula (1) is provided:

[0008]

[0009] Wherein:

[0010] R1 is selected from C1-C4 alkyl;

[0011] R4, R5, R6 and R7 are independently selected from halogen, C1-C4 alkyl, C1-C4 alkoxy, trifluoromethoxy.

[0012] Preferably, the compound is the compound represented by formula (II):

[0013]

[0014] Wherein, R4, R5, R6 and R7 are independently selected from fluorine, chlorine, bromine, iodine; methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, tert-butyl; methoxy, ethoxy, propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, trifluoromethoxy.

[0015] As a preference, the compound provided by this invention is:

[0016] T1: Methyl N'-(5-fluoro-1-allyl-2-oxoindoline-3-ylidene)hydrazine dithiocarboxylate

[0017] T2: Methyl N'-(6-fluoro-1-allyl-2-oxoindoline-3-ylidene)hydrazinecarbodithioate

[0018] T3: Methyl N'-(7-fluoro-1-allyl-2-oxoindoline-3-ylidene)hydrazinecarbodithioate

[0019] T4: Methyl N'-(4,7-difluoro-1-allyl-2-oxoindoline-3-ylidene)hydrazinecarbodithioate

[0020] T5: Methyl N'-(5-chloro-1-allyl-2-oxoindoline-3-ylidene)hydrazinecarbodithioate

[0021] T6: Methyl N'-(6-chloro-1-allyl-2-oxoindoline-3-ylidene)hydrazinecarbodithioate

[0022] T7: Methyl N'-(7-chloro-1-allyl-2-oxoindoline-3-ylidene)hydrazinecarbodithioate

[0023] T8: Methyl N'-(5-bromo-1-allyl-2-oxoindoline-3-ylidene)hydrazinecarbodithioate

[0024] T9: Methyl N'-(5-iodo-1-allyl-2-oxoindoline-3-ylidene)hydrazinecarbodithioate

[0025] T10: Methyl N'-(7-methyl-1-allyl-2-oxoindoline-3-ylidene)hydrazinecarbodithioate

[0026] T11: Methyl N'-(5-isopropyl-1-allyl-2-oxoindoline-3-ylidene)hydrazinecarbodithioate

[0027] T12: Methyl N'-(5-methoxy-1-allyl-2-oxoindoline-3-ylidene)hydrazinecarbodithioate

[0028] T13: Methyl N'-(6-methoxy-1-allyl-2-oxoindoline-3-ylidene)hydrazinecarbodithioate

[0029] T14: Methyl N'-(5-trifluoromethoxy-1-allyl-2-oxoindoline-3-ylidene)hydrazinecarbodithioate

[0030] T15: Methyl N'-(6-trifluoromethoxy-1-allyl-2-oxoindoline-3-ylidene)hydrazinecarbodithioate.

[0031] In a second aspect, the present invention provides a method for preparing the compound, which comprises the following steps:

[0032]

[0033] Among them, the definitions of R4, R5, R6 and R7 in formulas (III) and (IV) are the same as those in the compound of formula (I).

[0034] (1) At 25 °C, under the catalysis of potassium carbonate, 1 equivalent of indoline-2,3-dione and 1 - 1.2 equivalents of 3-bromopropene were added to a round-bottom flask equipped with a magnetic stirrer. 2 equivalents of potassium carbonate were added, and then the solvent N,N-dimethylformamide was added dropwise. After stirring overnight at 25 °C, water was added, and a solid precipitated. The solid was filtered by suction, repeatedly washed with water 3 times, and the obtained filter cake was dried under reduced pressure at 50 °C to obtain intermediate IV.

[0035] (2) 1 equivalent of intermediate IV was put into a round-bottom flask equipped with a magnetic stirrer, 1 - 1.1 equivalents of hydrazine dithiocarbonate were added, a small amount of glacial acetic acid was added, and then methanol was added. After stirring at 25 °C for 10 hours, the mixture was filtered by suction, repeatedly washed with methanol 3 times, and the obtained filter cake was dried under reduced pressure at 50 °C to obtain the final product I.

[0036] In a third aspect, the present invention provides a pharmaceutical composition, which contains the above-mentioned compound or its stereoisomer, or its pharmaceutically acceptable salt, and one or more pharmaceutically acceptable carriers, diluents, adjuvants, vehicles or excipients.

[0037] In a fourth aspect, the present invention provides the use of the above-mentioned compound or its pharmaceutically acceptable salt in the preparation of a New Delhi metallo-β-lactamase 1 inhibitor, and provides the use of methyl N'-(1-allyl-2-oxoindoline-3-ylidene)hydrazine dithiocarbonate or its pharmaceutically acceptable salt in the preparation of a New Delhi metallo-β-lactamase 1 inhibitor.

[0038] In a fifth aspect, the present invention provides a combined drug, which contains all of the above-mentioned compounds or their stereoisomers, or their pharmaceutically acceptable salts, and one or more pharmaceutically acceptable carriers, diluents, adjuvants, vehicles or excipients and β-lactam antibiotics.

[0039] Preferably, the β-lactam antibiotics are selected from one or more of penicillins, cephalosporins, cephamycins, thienamycins, monocyclic β-lactams and carbapenems: ertapenem, meropenem, doripenem, biapenem, panipenem, ticarcillin, ampicillin, amoxicillin, carbenicillin, piperacillin, azlocillin, mezlocillin, ticarcillin, cefoperazone, cefotaxime, ceftriaxone, cefepime, ceftolozane and ceftazidime.

[0040] Sixth aspect, the present invention provides the use of the above-mentioned combined drugs in the preparation of drugs against NDM-1 enzyme-producing bacteria or superbugs

[0041] Preferably, the NDM-1 enzyme-producing bacteria or superbugs are selected from one or more of Escherichia coli, Klebsiella pneumoniae, Acinetobacter baumannii, Enterobacter cloacae, Proteus, Citrobacter freundii, Klebsiella oxytoca, Morganella morganii, and Providencia

[0042] Compared with the prior art, the present invention has the following beneficial effects:

[0043] The experiment first found that N-allyl-substituted indoline-2-one-3-hydrazine dithiocarboxylate derivatives have obvious inhibitory activity against New Delhi metallo-β-lactamase 1 (NDM-1). When combined with meropenem, it can restore the sensitivity of NDM-1 enzyme-producing Escherichia coli ATCC BAA-2469 to meropenem. Since the structure type is different from the reported NDM-1 inhibitors, it has the value of in-depth research and the potential for clinical application Specific embodiments

[0044] The present invention will be further described below in conjunction with specific embodiments. However, the embodiments are only used to illustrate the present invention, rather than to limit the present invention. The experimental methods used in the embodiments are all conventional methods unless otherwise specified; the materials, reagents, etc. used are reagents and materials that can be obtained from commercial channels unless otherwise specified

[0045] I. Preparation and detection of new compounds

[0046] Example 1: Methyl N'-(5-fluoro-1-allyl-2-oxoindoline-3-ylidene)hydrazine dithiocarboxylate (T1)

[0047]

[0048] Add 5-fluoroindole-2,3-dione (500 mg, 1 eq) and 3-bromopropene (1.1 eq) to a 100 mL round-bottom flask equipped with a magnetic stirrer, add potassium carbonate (2 eq), and then dropwise add 10 mL of the solvent N,N-dimethylformamide. After stirring overnight at 25 °C, add 50 mL of water, and a solid precipitates. Filter by suction, and wash the solid repeatedly with water 3 times. The obtained filter cake is dried under reduced pressure at 50 °C to obtain the intermediate 5-fluoro-1-allyl-indole-2,3-dione (yield: 82%)

[0049] The obtained 5-fluoro-1-allyl-1H-indole-2,3-dione (509 mg) was placed into a 100 mL round-bottom flask equipped with a magnetic stir bar. Methyl hydrazinecarbodithioate (1.02 eq) was added, 2 μL of glacial acetic acid was added, and then 8 mL of methanol was added. After stirring at 25 °C for 10 hours, filtration was carried out, and the residue was repeatedly washed with methanol three times. The obtained filter cake was dried under reduced pressure at 50 °C to obtain 660 mg of the target product methyl N'-(5-fluoro-1-allyl-2-oxoindoline-3-ylidene)hydrazinecarbodithioate.

[0050] Orange-yellow solid, yield 86%. ESI-MS (m / z): 310.40 [M+H] + . 1 H NMR (500 MHz, Chloroform-d) δ 13.77 (s, 1H), 7.46–7.38 (m, 1H), 7.10–7.02 (m, 1H), 6.81 (dd, J = 8.6, 3.8 Hz, 1H), 5.83 (m, J = 10.7, 5.1 Hz, 1H), 5.31–5.24 (m, 2H), 4.38 (d, J = 4.8 Hz, 2H), 2.67 (s, 3H).

[0051] Example 2: Methyl N'-(6-fluoro-1-allyl-2-oxoindoline-3-ylidene)hydrazinecarbodithioate (T2)

[0052]

[0053] Using 6-fluoroindole-2,3-dione as the starting material, the synthesis method was the same as that in Example 1.

[0054] Orange-yellow solid, yield 74%. ESI-MS (m / z): 310.47 [M+H] + . 1 H NMR (500 MHz, Chloroform-d) δ 13.67 (s, 1H), 7.65 (dd, J = 8.3, 5.5 Hz, 1H), 6.91–6.68 (m, 1H), 6.61 (dd, J = 8.7, 2.2 Hz, 1H), 5.88–5.78 (m, 1H), 5.28 (t, J = 13.1 Hz, 2H), 4.39–4.34 (m, 2H), 2.65 (s, 3H).

[0055] Example 3: Methyl N'-(7-fluoro-1-allyl-2-oxoindoline-3-ylidene)hydrazinecarbodithioate (T3)

[0056]

[0057] Using 7-fluoroindole-2,3-dione as the starting material, the synthesis method is the same as that in Example 1.

[0058] Orange-yellow solid, yield 78%. ESI-MS (m / z): 308.55 [M-H]. 1 H NMR (500 MHz, Chloroform-d) δ 13.76 (s, 1H), 7.51 (d, J = 7.3 Hz, 1H), 7.19–6.99 (m, 2H), 5.92 (m, J = 10.7, 5.1 Hz, 1H), 5.48–5.08 (m, 2H), 4.51 (d, J = 5.4 Hz, 2H), 2.67 (s, 3H).

[0059] Example 4: Methyl N'-(4,7-difluoro-1-allyl-2-oxoindoline-3-ylidene)hydrazinecarbodithioate (T4)

[0060]

[0061] Using 4,7-difluoroindole-2,3-dione as the starting material, the synthesis method is the same as that in Example 1.

[0062] Orange-yellow solid, yield 53%. ESI-MS (m / z): 350.33 [M+Na] + . 1 H NMR (400 MHz, Chloroform-d) δ 13.76 (s, 1H), 7.06 (m, J = 10.5, 9.2, 4.0 Hz, 1H), 6.76 (m, J = 9.3, 8.3, 3.0 Hz, 1H), 5.95–5.86 (m, 1H), 5.27–5.21 (m, 2H), 4.50 (dd, J = 5.5, 1.5 Hz, 2H), 2.65 (s, 3H).

[0063] Example 5: Methyl N'-(5-chloro-1-allyl-2-oxoindoline-3-ylidene)hydrazinecarbodithioate (T5)

[0064]

[0065] Using 5-chloroindole-2,3-dione as the starting material, the synthesis method is the same as that in Example 1.

[0066] Orange-yellow solid, yield 74%. ESI-MS (m / z): 348.35 [M+Na] + . 11H NMR (500 MHz, Chloroform-d) δ 13.74 (s, 1H), 7.68 (d, J = 2.1 Hz, 1H), 7.34 (dd, J = 8.4, 2.1 Hz, 1H), 6.82 (d, J = 8.4 Hz, 1H), 5.83 (m, J = 10.7, 5.2 Hz, 1H), 5.37–5.18 (m, 2H), 4.38 (m, J = 5.4, 1.7 Hz, 2H), 2.68 (s, 3H).

[0067] Example 6: Methyl N'-(6-chloro-1-allyl-2-oxoindoline-3-ylidene)hydrazinecarbodithioate (T6)

[0068]

[0069] Using 6-chloroindole-2,3-dione as the starting material, the synthesis method is the same as that in Example 1.

[0070] Orange-yellow solid, yield 77%. ESI-MS (m / z): 326.60 [M+H] + . 1 1H NMR (500 MHz, Chloroform-d) δ 13.72 (s, 1H), 7.63 (d, J = 8.1 Hz, 1H), 7.11 (dd, J = 8.0, 1.8 Hz, 1H), 6.88 (d, J = 1.8 Hz, 1H), 5.84 (m, J = 10.6, 5.1 Hz, 1H), 5.47–5.15 (m, 2H), 4.37 (m, J = 3.4, 1.8 Hz, 2H), 2.67 (s, 3H).

[0071] Example 7: Methyl N'-(7-chloro-1-allyl-2-oxoindoline-3-ylidene)hydrazinecarbodithioate (T7)

[0072]

[0073] Using 7-chloroindole-2,3-dione as the starting material, the synthesis method is the same as that in Example 1.

[0074] Orange-yellow solid, yield 65%. ESI-MS (m / z): 326.22 [M+H] + . 11H NMR (500 MHz, Chloroform-d) δ 13.75 (s, 1H), 7.64 (d, J = 7.4 Hz, 1H), 7.32 (d, J = 8.1 Hz, 1H), 7.06 (t, J = 7.8 Hz, 1H), 5.98 (m, J = 15.9, 10.4, 5.1 Hz, 1H), 5.20 (dd, J = 20.7, 13.8 Hz, 2H), 4.78 (d, J = 4.9 Hz, 2H), 2.67 (s, 3H).

[0075] Example 8: Methyl N'-(5-bromo-1-allyl-2-oxoindoline-3-ylidene)hydrazinecarbodithioate (T8)

[0076]

[0077] Using 5-bromoindole-2,3-dione as the starting material, the synthesis method is the same as that in Example 1.

[0078] Orange-yellow solid, yield 69%. ESI-MS (m / z): 392.42 [M+Na] + . 1 1H NMR (500 MHz, Chloroform-d) δ 13.72 (s, 1H), 7.82 (d, J = 2.0 Hz, 1H), 7.48 (dd, J = 8.4, 2.0 Hz, 1H), 6.77 (d, J = 8.4 Hz, 1H), 5.83 (m, J = 16.1, 10.6, 5.4 Hz, 1H), 5.31–5.21 (m, 2H), 4.38 (d, J = 5.2 Hz, 2H), 2.68 (s, 3H).

[0079] Example 9: Methyl N'-(5-iodo-1-allyl-2-oxoindoline-3-ylidene)hydrazinecarbodithioate (T9)

[0080]

[0081] Using 5-iodoindole-2,3-dione as the starting material, the synthesis method is the same as that in Example 1.

[0082] Orange-yellow solid, yield 85%. ESI-MS (m / z): 440.45 [M+Na] + . 11H NMR (500 MHz, Chloroform-d) δ 13.77 (s, 1H), 7.53 (dd, J = 7.9, 1.4 Hz, 1H), 7.45 (d, J = 7.9 Hz, 1H), 7.30 (s, 1H), 5.87 (m, J = 10.6, 5.1 Hz, 1H), 5.32 (dd, J = 19.2, 13.7 Hz, 2H), 4.40 (m, J = 5.5, 1.7 Hz, 2H), 2.71 (s, 3H).

[0083] Example 10: Methyl N'-(7-methyl-1-allyl-2-oxoindoline-3-ylidene)hydrazinecarbodithioate (T10)

[0084]

[0085] Using 7-methylindole-2,3-dione as the starting material, the synthesis method is the same as that in Example 1.

[0086] Orange-yellow solid, yield 78%. ESI-MS (m / z): 328.45 [M+Na] + . 1 1H NMR (500 MHz, Chloroform-d) δ 13.84 (s, 1H), 7.59 (d, J = 7.4 Hz, 1H), 7.13 (d, J = 7.7 Hz, 1H), 7.02 (t, J = 7.6 Hz, 1H), 5.97 (m, J = 18.2, 10.5, 4.1 Hz, 1H), 5.23 (d, J = 10.7 Hz, 1H), 5.14–5.08 (m, 1H), 4.65–4.58 (m, 2H), 2.67 (s, 3H), 2.49 (s, 3H).

[0087] Example 11: Methyl N'-(5-isopropyl-1-allyl-2-oxoindoline-3-ylidene)hydrazinecarbodithioate (T11)

[0088]

[0089] Using 5-isopropylindole-2,3-dione as the starting material, the synthesis method is the same as that in Example 1.

[0090] Orange-yellow solid, yield 82%. ESI-MS (m / z): 334.70 [M+H] + . 11H NMR (500 MHz, Chloroform-d) δ 13.86 (s, 1H), 7.56 (s, 1H), 7.23 (dd, J = 8.2, 1.8 Hz, 2H), 6.80 (d, J = 8.1 Hz, 1H), 5.84 (m, J = 10.6, 5.2 Hz, 1H), 5.27 (dd, J = 13.6, 3.8 Hz, 2H), 4.38–4.34 (m, 2H), 2.98–2.85 (m, 1H), 2.69 (s, 3H), 1.27 (d, J = 6.9 Hz, 6H).

[0091] Example 12: Methyl N'-(5-methoxy-1-allyl-2-oxoindoline-3-ylidene)hydrazinecarbodithioate (T12)

[0092]

[0093] Using 5-methoxyindole-2,3-dione as the starting material, the synthesis method is the same as that of Example 1.

[0094] Orange-yellow solid, yield 65%. ESI-MS (m / z): 382.60 [M+Na] + . 1 1H NMR (500 MHz, Chloroform-d) δ 13.90 (s, 1H), 7.30 (dd, J = 13.9, 6.5 Hz, 1H), 6.89–6.76 (m, 1H), 5.96–5.80 (m, 1H), 5.31 (m, J = 7.3 Hz, 2H), 4.40 (t, J = 6.6 Hz, 2H), 3.89 (d, J = 6.0 Hz, 3H), 2.73 (d, J = 6.0 Hz, 3H).

[0095] Example 13: Methyl N'-(6-methoxy-1-allyl-2-oxoindoline-3-ylidene)hydrazinecarbodithioate (T13)

[0096]

[0097] Using 6-methoxyindole-2,3-dione as the starting material, the synthesis method is the same as that of Example 1.

[0098] Orange-yellow solid, yield 82%. ESI-MS (m / z): 344.33 [M+23] + . 11H NMR (500 MHz, Chloroform-d) δ 13.73 (s, 1H), 7.61 (d, J = 8.4 Hz, 1H), 6.61 (dd, J = 8.4, 2.2 Hz, 1H), 6.42 (d, J = 2.3 Hz, 1H), 5.84 (m, J = 10.7, 5.2 Hz, 1H), 5.33–5.15 (m, 2H), 4.35 (d, J = 5.2 Hz, 2H), 3.86 (s, 3H), 2.66 (s, 3H).

[0099] Example 14: Methyl N'-(5-trifluoromethoxy-1-allyl-2-oxoindoline-3-ylidene)hydrazinecarbodithioate (T14)

[0100]

[0101] Using 5-trifluoromethoxyindole-2,3-dione as the starting material, the synthesis method is the same as that in Example 1.

[0102] Orange-yellow solid, yield 82%. ESI-MS (m / z): 376.61 [M+H] + . 1 1H NMR (500 MHz, Chloroform-d) δ 13.75 (s, 1H), 7.57 (s, 1H), 7.23 (s, 1H), 6.87 (dd, J = 20.7, 8.6 Hz, 1H), 5.84 (m, J = 15.9, 10.4, 5.3 Hz, 1H), 5.29 (t, J = 13.1 Hz, 2H), 4.41 (s, 2H), 2.68 (s, 3H).

[0103] Example 15: Methyl N'-(6-trifluoromethoxy-1-allyl-2-oxoindoline-3-ylidene)hydrazinecarbodithioate (T15)

[0104]

[0105] Using 6-trifluoromethoxyindole-2,3-dione as the starting material, the synthesis method is the same as that in Example 1.

[0106] Orange-yellow solid, yield 86%. ESI-MS (m / z): 376.67 [M+H] + . 11H NMR (500 MHz, Chloroform-d) δ 13.71 (s, 1H), 7.72 (d, J = 8.3 Hz, 1H), 6.98 (d, J = 8.3 Hz, 1H), 6.74 (s, 1H), 5.84 (m, J = 16.0, 10.4, 5.4 Hz, 1H), 5.31 (d, J = 11.0 Hz, 2H), 4.44–4.32 (m, 2H), 2.68 (s, 3H).

[0107] II. Obtaining and Preparation of the Compound Z1 with a Disclosed Structure

[0108]

[0109] Methyl N'-(1-allyl-2-oxoindoline-3-ylidene)hydrazinecarbodithioate (CAS No.: 364055-05-0)

[0110] Example 1: Purchased from Compound Handling B.V. (Trade name: Specs), company address: Bleiswijkseweg 55, 2712 PB Zoetermeer, The Netherlands. The purchase number is: AG-670 / 33914015

[0111] Example 2: Synthesis and Structural Characterization

[0112]

[0113] Indole-2,3-dione (500 mg, 1 eq) and 3-bromopropene (1.1 eq) were added to a 100 ml round-bottom flask equipped with a magnetic stir bar. Potassium carbonate (2 eq) was added, and then 10 mL of the solvent N,N-dimethylformamide was added dropwise. After stirring overnight at 25 °C, 50 mL of water was added, and a solid precipitated. The solid was filtered by suction, washed repeatedly with water 3 times, and the obtained filter cake was dried under reduced pressure at 50 °C to obtain 553 mg of the intermediate 1-allylindole-2,3-dione (yield: 87%).

[0114] The obtained 1-allyl-1H-indole-2,3-dione was weighed and put into a 100 mL round-bottom flask equipped with a magnetic stir bar. Methyl hydrazinecarbodithioate (1.02 eq) was added, 2 μL of glacial acetic acid was added, and then 8 mL of methanol was added. After stirring at 25 °C for 10 hours, the mixture was filtered by suction, washed repeatedly with methanol 3 times, and the obtained filter cake was dried under reduced pressure at 50 °C to obtain 646 mg of the target product methyl N'-(1-allyl-2-oxoindoline-3-ylidene)hydrazinecarbodithioate.

[0115] Orange-yellow solid, yield 75%. ESI-MS (m / z): 314.31 [M+Na]+ . 1 1H NMR (500 MHz, DMSO-d6) δ 13.87 (s, 1H), 7.59 (d, J = 7.5 Hz, 1H), 7.47 (t, J = 7.8 Hz, 1H), 7.22–7.06 (m, 2H), 5.87 (td, J = 10.8, 5.1 Hz, 1H), 5.24 (dd, J = 34.8, 13.8 Hz, 2H), 4.38 (d, J = 5.1 Hz, 2H), 2.63 (s, 3H).

[0116] III. Biological Activity Assay

[0117] Experimental Example 1: Inhibitory Activity Test of the Compound against NDM-1 Enzyme

[0118] 1. Experimental Materials and Instruments

[0119] · 96-well UV analysis plate: CORNING;

[0120] · N-2-Hydroxyethylpiperazine-N'-2-ethanesulfonic acid free acid (HEPES FREE ACID): Amresco, USA;

[0121] · DMSO: Beijing Innochem Science & Technology Co., Ltd.;

[0122] · Meropenem trihydrate: Shanghai Bidepharm Co., Ltd.;

[0123] · 2,6-Pyridinedicarboxylic acid (DPA): Shanghai Bidepharm Co., Ltd.;

[0124] · Microplate reader: PerkinElmer Enspire 2300 Multiabel Reader

[0125] 2. Final Experimental System

[0126] · Volume 200 μL, NDM-1 enzyme concentration 1 nM, meropenem concentration 250 μM, 50 mM HEPES (pH = 7.35), 250 mM NaCl, 10 μg / mL BSA, 25 °C (DMSO content ≤ 1%).

[0127] 3. Experimental Method

[0128] · Serial dilution, final concentrations of the inhibitor and the positive compound DPA were set as: 200 μM, 100 μM, 50 μM, 25 μM, 12.50 μM, 6.25 μM, 3.13 μM, 1.56 μM, 0.78 μM, 0.39 μM, 0.20 μM, 0.10 μM (12 concentration gradients);

[0129] · Incubate the inhibitor with NDM-1 enzyme (excluding all substrates) for 10 min. Add 100 μL of substrate, mix well to initiate the reaction;

[0130] · Monitor the reaction. Measure the absorbance of the system at 300 nm every 1 min and continuously detect for 30 min at 25 °C. Calculate the enzyme activity at each inhibitor concentration and fit the IC50 using GraphPad Prism 8 software (the fitted curve serves as the original data);

[0131] · Set up three replicate wells in this experiment. Also set up a positive control group (enzyme, substrate, DPA), a negative control group (enzyme, substrate, DMSO), a blank control group (substrate, DMSO), and an experimental group (enzyme, substrate, inhibitor).

[0132] 4. The inhibitory activity of the compound against NDM-1 is shown in Table 1.

[0133] Experimental Example 2: In vitro combined antibacterial activity test of the compound

[0134] 1. Test strains and their culture methods

[0135] · Test strains

[0136] Escherichia coli ATCC BAA-2469 producing NDM-1 enzyme.

[0137] · Culture medium and culture conditions

[0138] CAMHB broth, incubate at 35 - 37 °C for 18 - 20 h and observe the results.

[0139] · Preparation of the culture medium

[0140] Preparation of CAMHB broth: Weigh an appropriate amount of CAMHB dry powder, add an appropriate amount of pure water, stir to dissolve, autoclave at 121 °C for 20 min, and cool to room temperature for standby. Prepare and use immediately.

[0141] · Positive control

[0142] VNRX-5133: Purchased from MedChemExpress (MCE).

[0143] 2. Test method

[0144] · Determine the minimum inhibitory concentration of meropenem against the test strain. Set the final concentration of meropenem to: 128 μg / ml, 64 μg / mL, 32 μg / mL, 16 μg / mL, 8 μg / mL, 4 μg / mL, 2 μg / mL, 1 μg / mL, 0.5 μg / mL, 0.25 μg / mL, 0.125 μg / mL, 0.0625 μg / mL.

[0145] · Determine the minimum inhibitory concentration (MIC) of the compound, and set the final concentrations to: 32 μg / mL, 16 μg / mL, 8 μg / mL, 4 μg / mL, 2 μg / mL, 1 μg / mL.

[0146] · Determine the MIC when meropenem is combined with the compound (32 μg / mL, 16 μg / mL, 8 μg / mL, 4 μg / mL, 2 μg / mL).

[0147] 3. The antibacterial activity of the compound combined with meropenem is shown in Table 1.

[0148] Table 1. Inhibitory activity of the compound against NDM-1

[0149]

[0150]

[0151]

Claims

1. A compound of formula (I) or a pharmaceutically acceptable salt thereof, wherein: R1 is selected from C1-C4 alkyl; R4, R5, R6 and R7 are independently selected from halogen, C1-C4 alkyl, C1-C4 alkoxy, trifluoromethoxy.

2. The compound or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that, The compound is the compound shown in formula (II): wherein, R4, R5, R6 and R7 are independently selected from fluorine, chlorine, bromine, iodine; methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, tert-butyl; methoxy, ethoxy, propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, trifluoromethoxy.

3. The following compound or a pharmaceutically acceptable salt thereof, characterized in that, The compound is selected from the following groups: T1: Methyl N'-(5-fluoro-1-allyl-2-oxoindoline-3-ylidene)hydrazinecarbodithioate T2: Methyl N'-(6-fluoro-1-allyl-2-oxoindoline-3-ylidene)hydrazinecarbodithioate T3: Methyl N'-(7-fluoro-1-allyl-2-oxoindoline-3-ylidene)hydrazinecarbodithioate T4: Methyl N'-(4,7-difluoro-1-allyl-2-oxoindoline-3-ylidene)hydrazinecarbodithioate T5: Methyl N'-(5-chloro-1-allyl-2-oxoindoline-3-ylidene)hydrazinecarbodithioate T6: Methyl N'-(6-chloro-1-allyl-2-oxoindoline-3-ylidene)hydrazinecarbodithioate T7: Methyl N'-(7-chloro-1-allyl-2-oxoindoline-3-ylidene)hydrazinecarbodithioate T8: Methyl N'-(5-bromo-1-allyl-2-oxoindoline-3-ylidene)hydrazinecarbodithioate T9: Methyl N'-(5-iodo-1-allyl-2-oxoindoline-3-ylidene)hydrazinecarbodithioate T10: Methyl N'-(7-methyl-1-allyl-2-oxoindoline-3-ylidene)hydrazinecarbodithioate T11: Methyl N'-(5-isopropyl-1-allyl-2-oxoindoline-3-ylidene)hydrazinecarbodithioate T12: Methyl N'-(5-methoxy-1-allyl-2-oxoindoline-3-ylidene)hydrazinecarbodithioate T13: Methyl N'-(6-methoxy-1-allyl-2-oxoindoline-3-ylidene)hydrazinecarbodithioate T14: Methyl N'-(5-trifluoromethoxy-1-allyl-2-oxoindoline-3-ylidene)hydrazinecarbodithioate T15: Methyl N'-(6-trifluoromethoxy-1-allyl-2-oxoindoline-3-ylidene)hydrazinecarbodithioate.

4. A method for preparing the compound according to claim 1 or 2, characterized in that, Comprising the following steps: wherein, the definitions of R1, R4, R5, R6 and R7 in formulas (III) and (IV) are as defined in the compound according to claim 1 or 2.

5. A pharmaceutical composition, characterized in that, It comprises the compound according to any one of claims 1-3 or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers, diluents, adjuvants, vehicles or excipients.

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

7. Use of compound Z1 or a pharmaceutically acceptable salt thereof in the preparation of a New Delhi metallo-β-lactamase 1 inhibitor: Z1: Methyl N'-(1 - allyl - 2 - oxoindoline - 3 - ylidene)hydrazinecarbodithioate.

8. A combined drug, characterized in that, It comprises the compound according to any one of claims 1-3 or a pharmaceutically acceptable salt thereof in combination with a β-lactam antibiotic.

9. The combined drug according to claim 8, wherein the β-lactam antibiotic is selected from one or more of penicillins, cephalosporins, cephamycins, thienamycins, monocyclic β-lactams, and carbapenems.

10. The combined drug according to claim 8, wherein the β-lactam antibiotic is selected from one or more of ertapenem, meropenem, doripenem, biapenem, panipenem, ticarcillin, ampicillin, amoxicillin, carbenicillin, piperacillin, azlocillin, mezlocillin, cefoperazone, cefotaxime, ceftriaxone, cefepime, ceftolozane, and ceftazidime.

11. Use of the combined drug according to any one of claims 8-10 in the preparation of a drug against bacteria or superbugs producing NDM-1 enzyme.

12. The use according to claim 11, characterized in that, The bacteria or superbugs producing NDM-1 enzyme are selected from one or more of Escherichia coli, Klebsiella pneumoniae, Acinetobacter baumannii, Enterobacter cloacae, Proteus spp., Citrobacter freundii, Klebsiella oxytoca, Morganella morganii, and Providencia spp.

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