Chlorinated compound and application thereof as anti-mycobacterium tuberculosis medicine

Abstract

The invention relates to a chlorinated compound and application thereof as an anti-mycobacterium tuberculosis drug. The invention provides an anti-Mycobacterium tuberculosis infection agent, which is characterized in that the anti-Mycobacterium tuberculosis infection agent takes a compound shown as a formula I and / or a pharmaceutically acceptable salt thereof as an active ingredient, and is used for treating infectious diseases caused by Mycobacterium tuberculosis H37Ra.

Description

Technical Field

[0001] The invention belongs to the field of medicine, and in particular relates to a class of chlorinated compounds and their application as anti-tuberculosis drugs. Background Art

[0002] Tuberculosis is a chronic, infectious disease that seriously threatens human life and health. It ranks second in mortality among infectious diseases worldwide and has become an important public health issue worldwide (Telenti, A., et al, Lancet. 1993, 341, 647–650). According to data released by WHO's "Global Tuberculosis Report 2023", there are about 10 million new cases of tuberculosis patients worldwide each year, of which 1.3 million to 1.5 million will die from tuberculosis (World Health Organization, 2023); so far, tuberculosis is still the leading cause of death from infectious diseases worldwide (Urban, M., et al, Eur. J. Med. Chem. 2021, 212, 113139.). Isoniazid and rifampicin are the main clinical first-line drugs for the treatment of tuberculosis, but as the dosage of patients increases and the treatment cycle is prolonged during the treatment process, the side effects of the drugs increase and the drug resistance of Mycobacterium tuberculosis becomes increasingly obvious (Singh, R., et al, J. Appl. Microbiol. 2019, 128, 1547-1567.); the emergence of multi- / extensively drug-resistant Mycobacterium tuberculosis has made the global anti-tuberculosis situation increasingly severe (Monde, N., et al, Antibiotics. 2023, 12, 166.). The research on anti-tuberculosis drugs requires large investment and a long research cycle. There has been no breakthrough since the advent of rifampicin in 1963 (Dartois, VA, et al, Nat. Rev. Microbiol. 2022, 20, 685–701.). The pathogen Mycobacterium tuberculosis has strong pathogenicity, slow growth rate, harsh research conditions (biosafety level 3 or above laboratories), uncertainty of important metabolic targets in the in vivo environment, and lack of chemical diversity in sample libraries used for screening, which are all important reasons for the slow development of anti-tuberculosis drugs. Summary of the invention

[0003] The object of the present invention is to provide a chlorinated compound and its use as an anti-tuberculosis bacillus drug, which can meet the above-mentioned requirements of the prior art.

[0004] The present invention provides a chloro compound 1-3 of formula I or a pharmaceutically acceptable salt thereof or a solvate of the salt thereof, characterized in that the compound has the following structure:

[0005] Wherein, R is phenyl, o-chlorostyryl, m-chlorostyryl, specifically,

[0006] The compound of the present invention has strong inhibitory activity on mycobacterium tuberculosis, which is equivalent to positive drugs rifampicin and isoniazid, can be used as a drug for treating mycobacterium tuberculosis infection, and has broad application prospects.

[0007] In another embodiment of the present invention, an anti-Mycobacterium tuberculosis drug is provided, characterized in that it comprises compounds 1-3 of formula I or pharmaceutically acceptable salts thereof as an active ingredient.

[0008] The term "pharmaceutically acceptable salt" in the present invention refers to non-toxic addition salts of inorganic or organic acids and / or bases; see "Salt selection for basic drugs", Int. J. Pharm. 1986, 33, 201-217. DETAILED DESCRIPTION

[0009] In order to facilitate further understanding of the present invention, the following examples are provided to explain it in more detail; however, these examples are only for a better understanding of the invention and are not intended to limit the scope or implementation principles of the present invention. The implementation methods of the present invention are not limited to the following contents.

[0010] Example

[0011] Example 1: 2-chloro-6-(5-hydroxy-3,7,8-trimethoxy-4-oxo-4H-chromen-2-yl)phenyl benzoate (Compound 1)

[0012]

[0013] Compound 2-(3-chloro-2-hydroxyphenyl)-5-hydroxy-3,7,8-trimethoxy-4H-chromen-4-one (hereinafter referred to as "Compound A") (50.0 mg, 0.13 mmol) was dissolved in dichloromethane (15 mL), and reacted with benzoic acid reagent (1-2 eq) in the presence of 4-dimethylaminopyridine (63.5 mg, 0.52 mmol, 4 eq) and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (136.1 mg, 0.71 mmol, 4 eq); the reaction solution was stirred at 45°C for reaction; after the reaction was completed as detected by TLC, water was added to stop the reaction, the reaction solution was extracted three times with dichloromethane, the combined organic phase was concentrated under reduced pressure and evaporated to dryness; a yellow solid was obtained by normal phase silica gel column chromatography with a yield of 42%.

[0014] 1 H NMR(400MHz,DMSO-d6)δppm 12.18(1H,s),8.02(2H,d,J=7.3Hz),7.92(1H,dd,J=8.1,1.1Hz),7.81–7.78(1H,m),7.71(1H, t,J=7.4Hz),7.60–7.53(3H,overlapped),6.58(1H,s),3.87(3H,s),3.68(3H,s),3.63(3H,s). 13 C NMR(100MHz,DMSO-d6)δ178.6,163.4,159.0,156.9,154.0,148.8,145.2,139.9,135.0,133.3,130.7,13 0.3,130.3,129.6,129.6,128.8,128.3,128.1,127.9,126.5,105.3,96.6,61.4,61.0,57.1; HRESIMSm / z 483.0836[M+H] + (calcd for C 25 H 20 O8Cl + ,483.0841); melting point 150.1-150.7℃.

[0015] Example 2: 2-chloro-6-(5-hydroxy-3,7,8-trimethoxy-4-oxo-4H-chromen-2-yl)phenyl(E)-3-(2-chlorophenyl)acrylate (Compound 2)

[0016]

[0017] A yellow flaky solid was obtained by referring to the preparation method of Example 1; the yield was 50%; 1 H NMR(400MHz,acetone-d6)δppm 12.31(1H,s),8.16(1H,d,J=16.0Hz),7.89(1H,dd,J=7.7,1.4Hz),7.83–7.77(2H,overlapped),7.57 –7.37(4H, overlapped),6.80(1H,d,J=16.0Hz),6.47(1H,s),3.91(3H,s),3.81(3H,s),3.75(3H,s). 13C NMR(100MHz,acetone-d6)δ179.6,163.3,159.7,158.0,154.5,149.6,146.1,142.9,140.6,135.2,133.2,132.8 ,132.5,130.8,130.7,129.6,129.0,128.9,128.3,128.0,127.2,119.6,106.1,96.4,61.4,60.8,56.7; HRESIMS m / z 543.0602(calcd for C 27 H 21 O8Cl2 + ,543.0608); melting point 150.5-151.2℃.

[0018] Example 3: 2-chloro-6-(5-hydroxy-3,7,8-trimethoxy-4-oxo-4H-chromen-2-yl)phenyl(E)-3-(3-chlorophenyl)acrylate (Compound 3)

[0019]

[0020] A yellow flaky solid was obtained by referring to the preparation method of Example 1; the yield was 60%; 1 H NMR(400MHz,acetone-d6)δppm 12.30(1H,s),7.86–7.74(4H,overlapped),7.65(1H,m),7.55(1H,t,J=8.0Hz),7.48–7.41(2 H, overlapped), 6.80 (1H, d, J = 16.0Hz), 6.46 (1H, s), 3.92 (3H, s), 3.80 (3H, s), 3.78 (3H, s). 13 C NMR(100MHz,acetone-d6)δ178.9,162.7,159.1,157.4,153.9,148.9,145.7,145.5,139.9,136.2,134.5,132.5 ,130.6,130.5,130.0,128.9,128.2,128.2,127.2,127.0,126.5,117.8,105.4,95.7,60.8,60.1,56.0.HRESIMS m / z543.0600[M+H] + (calcd for C 27 H 21 O8Cl2 +,543.0608); melting point 153.5-154.3℃.

[0021] Example 4: Mycobacterium tuberculosis activity of compounds 1-3 of the present invention

[0022] The Mycobacterium tuberculosis activity test method of the compounds 1-3 of the present invention is carried out according to the CLSI (Clinical and Laboratory Standards Institute). Performance standards for antimicrobial usceptibility testing; nineteenth informational supplement. M100-S19 (Wayne, 2009).

[0023] The activity test results showed that compounds 1–3 showed strong inhibitory activity against Mycobacterium tuberculosis M. tuberculosis H37Ra (MIC, 0.72–1.18 μM), which was comparable to the positive drugs rifampicin (MIC, 0.17 μM) and isoniazid (MIC, 1.5 μM), and was improved compared to compound A (MIC, 2.64 μM).

[0024] Example 5: Other antibacterial activity tests of compounds 1-3 of the present invention

[0025] Test strains: Staphylococcus aureus, Pseudomonas aeruginosa.

[0026] Test method: Ciprofloxacin was used as the positive control and DMSO was used as the negative control. Three parallel experiments were set up. The two bacteria were cultured in LB medium at 37°C for 8 hours and then diluted to 10 6 cfu / mL, using 96-well plates for detection, 198μL bacterial solution and 2μL test sample per well, detection concentration is 25μg / mL. Determination of MIC value: Observe from the back of the 96-well plate with the naked eye, record the clear wells to determine whether the bacteria grow, and thus determine the inhibitory effect and MIC value.

[0027] Results: At a concentration of 25 μg / mL, compounds 1–3 of the present invention had no significant inhibitory activity against the two bacteria tested.

[0028] Example 6: Other antibacterial activity tests of compounds 1-3 of the present invention

[0029] Test strains: Aspergillus fumigatus, Penicillium italicum, Penicillium digitatum.

[0030] Test method: 100 μL PDB and 100 μL spore suspension were used as blank controls, and amphotericin B was used as a positive control. Three parallel experiments were set up. The fungus was inoculated on PDA medium and cultured at 28±1℃ for 7 days for activation. Sterile water was added, and the spores on the agar surface were gently scraped off. The spore suspension was placed in a sterilized 250mL conical flask, and the filtrate volume was finally made to reach 100mL, which was used as the mother solution. The mother solution concentration was prepared to 10 7 cfu / mL; use a 96-well plate for detection, add 100 μL PDB, 98 μL of the above spore suspension and 2 μL sample to each well; set the maximum evaluation concentration to 25 μg / mL, place in a 28°C constant temperature incubator and culture for 48 hours, observe the results, and take the samples with no precipitation wells (no mycelium growth) as the activity results.

[0031] Results: At a concentration of 25 μg / mL, compounds 1–3 of the present invention had no significant inhibitory activity against the three fungi tested.

[0032] Through in vitro screening of the activity against other pathogenic bacteria, combined with Example 4, the compounds of the present invention have strong activity in inhibiting Mycobacterium tuberculosis and have certain selectivity, and are ineffective against other fungi and bacteria.

[0033] Experiments show that the compound of the present application has a strong inhibitory activity against Mycobacterium tuberculosis, and is higher than compound A. It can be made into an anti-Mycobacterium tuberculosis infection drug and has broad application prospects.

Claims

1. A chloro compound of formula I or a pharmaceutically acceptable salt thereof or a solvate of its salt, characterized in that The compound has the following structure: Wherein, R is phenyl, o-chlorostyryl, m-chlorostyryl, specifically, 2. An anti-mycobacterium tuberculosis agent, characterized in that Contains the compound of formula I or a pharmaceutically acceptable salt thereof as claimed in claim 1 as an active ingredient.

3. Use of the compound of formula I or a pharmaceutically acceptable salt thereof according to claim 1 in the preparation of an anti-Mycobacterium tuberculosis agent.

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