CN112778297B - Benzothiazole compound and preparation method and application thereof - Google Patents
Benzothiazole compound and preparation method and application thereof Download PDFInfo
- Publication number
- CN112778297B CN112778297B CN201911079527.7A CN201911079527A CN112778297B CN 112778297 B CN112778297 B CN 112778297B CN 201911079527 A CN201911079527 A CN 201911079527A CN 112778297 B CN112778297 B CN 112778297B
- Authority
- CN
- China
- Prior art keywords
- compound
- benzothiazole
- pharmaceutically acceptable
- general formula
- acceptable salts
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/38—Heterocyclic compounds having sulfur as a ring hetero atom
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
- A61K31/425—Thiazoles
- A61K31/428—Thiazoles condensed with carbocyclic rings
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
- A61P31/06—Antibacterial agents for tuberculosis
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D409/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms
- C07D409/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms containing two hetero rings
- C07D409/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms containing two hetero rings linked by a chain containing hetero atoms as chain links
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
- C07D417/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings
- C07D417/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings linked by a chain containing hetero atoms as chain links
Abstract
The invention belongs to the field of medicines, and particularly relates to a benzothiazole compound, a preparation method and an anti-tuberculosis application thereof. The benzothiazole compound is shown as follows, and is a high-efficiency, low-toxicity and stable antituberculosis drug.
Description
Technical Field
The invention belongs to the field of medicines, and particularly relates to a benzothiazole compound, a preparation method and an anti-tuberculosis application thereof.
Background
Tuberculosis (TB) is the highest death rate disease in infectious diseases caused by single cause in the world, and the Tuberculosis is classified as a disease seriously endangering global public health by the world health organization. The existing antituberculosis first-line treatment drugs used clinically generally have the defects of long treatment period, great toxic and side effects and wide drug resistance, and cannot meet the curing requirements. Therefore, the research and development of new action mechanisms and antituberculosis drugs with novel frameworks are urgent.
TB is caused by Mycobacterium Tuberculosis (MTB). The cell wall plays a vital role in the survival and propagation of the bacterium, the prevention of the formation of the MTB cell wall becomes a main idea of the development of the current anti-tuberculosis drugs, and the anti-tuberculosis drugs such as isoniazid, ethambutol, cycloserine and the like are developed aiming at related pathways of cell wall synthesis. In MTB, arabinose is a synthetic raw material of arabinogalactan, an important component of cell walls. The synthesis of DPA (Decaprenyl-Phosphoryl-D-Arabidopsis), an important precursor of Arabinose synthesis, requires the participation of Decaprenyl Phosphoryl beta-D-2' -epimerase 1(DprE 1). After the activity of DprE1 is inhibited, the synthesis of DPA can be effectively blocked, the synthesis of arabinose can be further blocked, the MTB cell wall can not be normally synthesized, and the aim of killing MTB can be achieved. The DprE1 enzyme is only present in the pathogen MTB, and has no related homologous protein in human host cells, so that the DprE1 enzyme becomes a brand-new anti-tuberculosis target.
Inhibitors targeted to DprE1 can be broadly classified into two broad classes, covalent and non-covalent, depending on the mechanism of action. Most covalently bound inhibitors covalently bind to the Cys387 active site in DprE1, resulting in irreversible inactivation of DprE 1. However, a series of mutations in DprE1 at Cys387 (Cys387Ser, Cys387Gly, Cys387Ala, etc.) resulted in a dramatic increase in the resistance profile of MTB to covalent inhibitors. The advent of DprE1 non-covalently bound inhibitors has solved the disadvantages of covalently bound inhibitors represented by Benzothiazinones (BTZs). In 5 months 2013, the first inhibitor, TCA1(ethyl (2- (benzo [ d ] thiazole-2-carboxamide) thiophene-3-carbonyl) carbamate, which is non-covalently bound to Dpr E1, was found in CAS No.:864941-32-2), and the chemical structure of TCA1 is shown in WO2014/190199Al (Table A, Cpd ID 1), which shows good bactericidal activity against both replicative and non-replicative MTB types, which BTZs do not have. The TCA1 has excellent activity and unique action mechanism on MTB strains, so that the MTB strains become an excellent antitubercular lead compound, but the research on the reconstruction of the TCA1 structure is few at present, and in addition, the clinical antitubercular first-line treatment drugs rifampicin and the like generally have the defects of long treatment period, high toxic and side effects and wide drug resistance. Thus, there remains a need for highly effective, low toxicity, stable antitubercular drugs.
Disclosure of Invention
The emergence of the non-covalently bound inhibitor TCA1 targeting DprE1 provides a new direction for the treatment of drug-resistant MTB and the development of DprE1 inhibitors. The invention provides a benzothiazole TCA1 derivative, which has superior performances in the aspects of DprE1 activity inhibition, drug resistance and non-drug resistance MTB strain inhibition, cytotoxicity and the like, and can be used as an antituberculosis drug. The invention is a more efficient, low-toxicity and stable antitubercular drug unexpectedly discovered in the reasonable optimization of TCA 1.
Accordingly, an object of the present invention is to provide benzothiazole compounds of the following general formula (I) or pharmaceutically acceptable salts thereof.
Another object of the present invention is to provide a process for preparing benzothiazole compounds of the following general formula (I) or pharmaceutically acceptable salts thereof.
The invention also provides application of the benzothiazole compound with the general formula (I) or pharmaceutically acceptable salt thereof in preparing medicines.
The invention provides benzothiazole compounds with the following general formula (I) or pharmaceutically acceptable salts thereof,
wherein R1 may be:
preferably, R1 may be:
more preferably, R1 may be:
in a specific embodiment, the benzothiazole compounds of the general formula (I) or pharmaceutically acceptable salts thereof are the following compounds or pharmaceutically acceptable salts thereof:
in a most preferred embodiment, the benzothiazole compounds of formula (I) or pharmaceutically acceptable salts thereof are the following compounds or pharmaceutically acceptable salts thereof:
the present invention provides a process for preparing benzothiazole compounds of the general formula (I) or pharmaceutically acceptable salts thereof, which comprises the steps of:
(1) synthesis of Compound B: adding cyanoacetic acid, the R1 substituted methyl carbamate compound A and acetic anhydride into a reaction vessel, heating in an oil bath, carrying out Ar protection, after full reaction, pouring the reaction solution into ice water, extracting with ethyl acetate, washing with a saturated sodium bicarbonate solution once, drying and concentrating to obtain a compound B;
(2) synthesis of Compound C: adding the compound B, 2, 5-dihydroxy-1, 4-dithiane, methanol and morpholine into a reaction vessel, carrying out oil bath for 2 hours to completely react, directly passing through a column, and eluting to obtain a compound C;
(3) synthesis of Compound (I): adding 2-formic acid benzothiazole and a compound C into a reaction container, adding pyridine, stirring for dissolving, dropwise adding phosphorus oxychloride, after stirring fully, adding dichloromethane and water into the system, quickly stirring to separate an organic layer, washing with a saturated sodium bicarbonate solution once, directly passing through a short silica gel column, and eluting to obtain a compound (I); or adding 2-formic acid benzothiazole and dichloromethane into a reaction container, cooling to 0 ℃, dropwise adding oxalyl chloride, reacting for several hours, adding pyridine into the system, stirring for several minutes, adding a compound C, reacting for several hours, adding methanol into the system to quench the reaction, washing once with a saturated sodium bicarbonate solution, directly mixing the sample with a silica gel column, and eluting to obtain the compound (I).
In addition, the invention provides application of the benzothiazole compound with the general formula (I) or pharmaceutically acceptable salt thereof in preparing antituberculosis drugs.
Preferably, the use of a benzothiazole compound of general formula (I) or a pharmaceutically acceptable salt thereof in the manufacture of an anti-tubercular medicament as a DprE1 inhibitor.
Preferably, the use of a benzothiazole compound of general formula (I) or a pharmaceutically acceptable salt thereof for the manufacture of an antitubercular medicament as H37Rv inhibitor.
Drawings
FIG. 1 shows the compound TCA1 in deuterated DMSO according to the experimental examples of the present invention1H-NMR spectrum.
FIG. 2 is a mass spectrum of TCA1, a compound involved in the experimental examples of the present invention.
FIG. 3 is a graph showing the inhibition of Mycobacterium tuberculosis DprE1 by the compounds LZDT1 and TCA1 of the present invention.
FIG. 4 is a graph showing the bactericidal activity of the compounds LZDT1 and TCA1 against M.tuberculosis H37 Rv.
FIG. 5 is a toxicity curve of the compound LZDT1 of the present invention on human hepatoma cell HepG 2.
FIG. 6 is a toxicity curve of TCA1 on human hepatoma cell HepG 2.
Detailed Description
The present invention will be described below with reference to examples, but the present invention is not limited thereto. The experimental procedures shown in the following examples are conventional unless otherwise specified. The reagents and materials shown are all commercially available products.
Preparation example 1 preparation of the Compound LZDT1
Adding 2.3g of cyanoacetic acid, 3g of compound A1 (cyclopropyl methyl carbamate) and 5mL of acetic anhydride into a 25mL single-neck bottle, heating the mixture in an oil bath at 85-90 ℃, protecting the mixture with Ar, reacting for 3-4h for complete reaction, pouring the reaction solution into ice water, extracting with ethyl acetate, washing with a saturated sodium bicarbonate solution once, drying and concentrating to obtain 1.5g of a reddish brown solid compound B1.
To a 100mL single neck flask were added compound B11 g, 0.42g of 2, 5-dihydroxy-1, 4-dithiane, 12mL of methanol, and 0.53g of morpholine, oil bath 65 deg.C, 2h to complete the reaction, column pass directly, EA: PE ═ 1: 2 to obtain 210mg of the solid compound C1.
Adding 166mg of 2-formic acid benzothiazole and 8mL of dichloromethane into a 25mL two-neck flask, cooling at 0 ℃, dropwise adding 129mg of oxalyl chloride, reacting for 2 hours, adding 175mg of pyridine into the system, stirring for 10min, adding a compound C1212 mg, reacting for 3 hours, adding 2mL of methanol into the system, quenching the reaction, washing once with 10mL of saturated sodium bicarbonate solution, directly stirring the sample, and passing through a column, EA: PE ═ 1: 2 to obtain 35mg of a pure solid compound LZDT1 with the yield of 9.9%.
Or adding 60mg of 2-formic acid benzothiazole and 180.5 mg of a compound in a 25mL two-opening bottle, adding 1mL of pyridine, stirring for dissolving, dropwise adding 0.05mL of phosphorus oxychloride at the temperature of-5 ℃, stirring for 1 hour, adding 5mL of dichloromethane and 1mL of water in the system, quickly stirring, separating an organic layer, washing the organic layer once with a saturated sodium bicarbonate solution, drying and concentrating, directly mixing the sample with a silica gel column, EA: PE ═ 1: 2, 15mg of the pure compound LZDT1 is obtained in a yield of 15%.
1H NMR(400MHz,DMSO-d6)δ13.10(s,1H),10.93(s,1H),8.32(d,J＝7.9Hz,2H),7.81(d,J＝5.9Hz,1H),7.68(dt,J＝19.1,7.3Hz,2H),7.21(d,J＝5.8Hz,1H),4.03(d,J＝7.3Hz,2H),1.27–1.14(m,1H),0.59(h,J＝4.5Hz,2H),0.38(t,J＝4.8Hz,2H).MS(ESI)cacld.forC18H15N3O4S2 401.97[M+1]+.
Preparation example 2 preparation of the Compound LZDT2
Adding 2.7g of cyanoacetic acid, 3.9g of compound A2 (cyclobutylmethyl carbamate) and 8mL of acetic anhydride into a 50mL single-neck bottle, heating the mixture in an oil bath at 85-90 ℃, carrying out Ar protection, reacting for 3-4h for completely reacting, pouring the reaction liquid into ice water, extracting with ethyl acetate, washing with a saturated sodium bicarbonate solution once, drying and concentrating to obtain 2.2g of a reddish brown solid compound B2.
To a 100mL single neck flask were added compound B21 g, 0.39g of 2, 5-dihydroxy-1, 4-dithiane, 10mL of methanol, and 0.49g of morpholine, oil bath 65 deg.C, 2h to complete the reaction, column pass directly, EA: PE ═ 1: 2 to give 0.4g of solid compound C2.
Adding 74mg of 2-benzoic acid benzothiazole and a compound C2105 mg into a 25mL two-neck bottle, adding 1mL of pyridine, stirring, dissolving, dropwise adding 0.05mL of phosphorus oxychloride at 5 ℃, stirring for 1h, adding 5mL of dichloromethane and 1mL of water into the system, quickly stirring, separating an organic layer, washing the organic layer once with a saturated sodium bicarbonate solution, directly passing through a short silica gel column, EA: PE 1:3 gave 5mg of pure LZDT2 as a solid with a yield of 4.16%.
1H NMR(400MHz,Chloroform-d)δ13.03(s,1H),8.26(d,J＝8.2Hz,1H),7.99(d,J＝8.0Hz,1H),7.94(s,1H),7.60(t,J＝7.6Hz,1H),7.53(t,J＝7.5Hz,1H),7.13(d,J＝5.9Hz,1H),6.96(d,J＝5.8Hz,1H),4.26(d,J＝7.0Hz,2H),2.71(p,J＝7.4Hz,1H),2.15–2.08(m,2H),2.02–1.9(m,2H),1.88–1.78(m,2H).MS(ESI)cacld.for C19H17N3O4S2 416.1[M+1]+.
Preparation example 3 preparation of the Compound LZDT3
Adding 0.62g of cyanoacetic acid, 1g of compound A3 (cyclopentyl methyl carbamate) and 8mL of acetic anhydride into a 50mL single-neck bottle, heating the mixture in an oil bath at 85-90 ℃ under the protection of Ar, reacting for 3-4h for complete reaction, pouring the reaction solution into ice water, extracting with ethyl acetate, washing with saturated sodium bicarbonate once, drying and concentrating to obtain 1.5g of a white-like solid compound B3.
A100 mL single vial was charged with B31.34g, 2, 5-dihydroxy-1, 4-dithiane 0.485g, methanol 13mL and morpholine 0.61g in an oil bath at 65 deg.C for 2h to complete the reaction, column passed directly, EA: PE ═ 1: 2 to give 0.85g of solid compound C3.
Adding 16.7mg of 2-benzoic acid benzothiazole and a compound C325 mg into a 25mL two-neck bottle, adding 1mL of pyridine, stirring to dissolve, dropwise adding 0.05mL of phosphorus oxychloride at the temperature of-5 ℃, stirring for 1h, adding 5mL of dichloromethane and 1mL of water into the system, quickly stirring, separating an organic layer, washing the organic layer once with a saturated sodium bicarbonate solution, directly passing through a short silica gel column, EA: PE 1:3 to give 3mg of pure LZDT3 as a solid with a yield of 7.5%.
1H NMR(400MHz,Chloroform-d)δ13.03(s,1H),8.27(d,J＝8.2Hz,1H),8.00(d,J＝8.0Hz,1H),7.88(s,1H),7.65–7.51(m,2H),7.11(d,J＝6.0Hz,1H),6.96(d,J＝5.9Hz,1H),4.17(d,J＝7.2Hz,2H),2.30(p,J＝7.7Hz,1H),1.80(m,2H)1.64(dd,J＝12.8,7.3Hz,3H),1.26(m,3H).MS(ESI)cacld.for C20H19N3O4S2 430.1[M+1]+.
Preparation example 4 preparation of the Compound LZDT4
Adding 1.39g of cyanoacetic acid, 2.48g of a compound A4 (cyclohexyl methyl carbamate) and 8mL of acetic anhydride into a 50mL single-neck bottle, heating the mixture in an oil bath at 85-90 ℃, protecting the mixture with Ar, reacting for 3-4h completely, pouring the reaction solution into ice water, extracting with ethyl acetate, washing with saturated sodium bicarbonate once, drying and concentrating to obtain 1.2g of a reddish brown solid compound B4.
A100 mL single vial was charged with B40.957g, 0.325g 2, 5-dihydroxy-1, 4-dithiane, 10mL methanol and 0.41g morpholine in an oil bath at 65 deg.C for 2h to complete the reaction, column passed directly, EA: PE ═ 1: 2 to give 0.3g of solid compound C4.
Adding 68mg of 2-benzoic acid benzothiazole and a compound C4100 mg into a 25mL two-neck bottle, adding 1mL of pyridine, stirring and dissolving, dropwise adding 0.05mL of phosphorus oxychloride at 5 ℃, stirring for 1h, adding 5mL of dichloromethane and 1mL of water into the system, quickly stirring, separating an organic layer, washing the organic layer once with a saturated sodium bicarbonate solution, directly passing through a short silica gel column, EA: elution with PE 1:3 gave 7mg of pure compound LZDT4 as a solid in 4.45% yield.
1H NMR(400MHz，Chloroform-d)δ13.03(s，1H)，8.26(d，J＝8.2Hz，1H)，8.00(d，J＝8.0Hz，1H)，7.89(s，1H)，7.60(t，J＝7.7Hz，1H)，7.54(d，J＝7.6Hz，1H)，7.11(d，J＝5.9Hz，1H),6.96(d，J＝5.9Hz，1H)，4.10(d，J＝6.4Hz，2H)，1.84–1.73(m，9H)，1.25(m，2H)。MS(ESI)cacld.for C21H21N3O4S2 444.0[M+1]+.
Experimental example 1 inhibition experiment of Mycobacterium Tuberculosis (MTB) Dpr E1 with the compound of the present invention
Experiment: DprE1, Flavin Adenine Dinucleotide (FAD), horseradish peroxidase (HRP), Amplex Red were dissolved in 50mM glycylglycine buffer (pH 8.0, 200mM potassium glutamate, 0.002% Brij-35, 1% DMSO) to final concentrations of 300nM, 1. mu.M, 0.2. mu.M and 50. mu.M, respectively. Mix well and aspirate the appropriate volume into a 384 well blackboard (Corning, cat # 3573). The compound TCA1 was obtained from Life Chemicals Inc. (ethyl (2- (benzol [ d ])]thiazole-2-carboxamide) thiophene-3-carbonyl) carbamate, CAS No. 864941-32-2), which1The H-NMR spectrum and the mass spectrum are shown in FIGS. 1 and 2, and are the same in the following experimental examples. FIG. 1 shows compound TCA1 dissolved in deuterated DMSO1H-NMR spectrum. FIG. 2 is a mass spectrum of compound TCA 1. 0-50 mu M of the compound to be tested (including the compound LZDT1 and a positive control drug TCA1) after gradient dilution is added into each hole, each concentration is repeated for 2 times, and the hole added with the same volume of DMSO is used as a negative control. After incubation at 30 ℃ for 10min, farnesyl-phosphoryl-. beta. -D-ribofuranose (FPR) was added to a final concentration of 300. mu.M to maintain a final volume of 30. mu.L. After being mixed uniformly, fluorescence detection is carried out on a full-wavelength multifunctional microplate reader Tecan M200 by using a kinetic mode, wherein the wavelength of excitation light is lambda ex which is 530nm, and the wavelength of emission light is lambda em which is 595 nm. The inhibitory median concentration (IC) of TCA1 and the compound LZDT1 of the invention for DprE1 activity was determined by fitting the initial reaction rates and the corresponding concentrations of the test compounds (including the compound LZDT1 of the invention and the positive control drug TCA1)50)。
As a result: the results of the experiment are shown in FIG. 3. FIG. 3 is a graph showing the inhibition of Mycobacterium tuberculosis DprE1 by the compounds LZDT1 and TCA1 of the present invention. IC of compound LZDT1 of the present invention against Mycobacterium Tuberculosis (MTB) DprE150Is 0.0158 +/-0.0028 mu M and is obviously smaller than the IC of positive control drug TCA1 on Mycobacterium Tuberculosis (MTB) Dpr E150(0.0583±0.0099μM)。
And (4) conclusion: the experimental result shows that the compound LZDT1 with higher inhibitory activity to DprE1 of Mycobacterium Tuberculosis (MTB) is obtained by the invention.
Experimental example 2 inhibition experiment of Mycobacterium tuberculosis H37Rv with the Compound of the present invention
Experiment: A7H 9 medium for culturing Mycobacterium tuberculosis H37Rv strain was prepared. 0.94g of 7H9 medium powder was weighed, 0.4mL of glycerol and 0.1mL of Tween80 were added, the mixture was fully dissolved with double distilled water and the volume was adjusted to 180mL, and the mixture was sterilized at 121 ℃ for 10 min. Before use, 20mL of filter-sterilized growth-promoting agent OADC (0.255g NaCl, 1.5g BSA-V, 0.6g glucose, 0.016g sodium oleate, 0.0012g catalase, double distilled water to 30mL) was added.
The H37Rv strain was inoculated into a 250mL Erlenmeyer flask containing 40mL of 7H9 medium (final kanamycin concentration is 20. mu.g/mL), and cultured in a constant temperature shaker at 37 ℃ at 110rpm for 10 days to the middle logarithmic growth phase to obtain H37Rv strain liquid. It was diluted to OD with fresh 7H9 medium (OADC added)6000.05 to 0.06 (in this case,. about.10)6CFU/mL fungal concentration). Pipette 100. mu.L of the bacterial suspension into each well of a 96-well microplate using a multi-channel pipette. mu.L of the compound of the invention LZDT1(4mg/mL) dissolved in DMSO was added to each well and two-fold gradient dilutions were performed in sequence. DMSO was used as a negative control, rifampicin and TCA1 were used as positive controls, and each test plate was subsequently sealed with Parafilm sealing film and incubated in a 37 ℃ incubator for 7 days. 30 mu L of resazurin is added into each hole, incubation is continued at 37 ℃ for 24 hours, the growth of bacteria is observed under a microscope, and the light absorption value of 570nm is detected by using a multifunctional microplate reader. The final concentration of the compound of the present invention in wells in which the growth of H37Rv strain was inhibited by more than 90% (fluorescence value less than 90% of that of the negative control group) was defined as the Minimum Inhibitory Concentration (MIC) of the compound against H37Rv strain.
As a result: according to the change of the absorbance values corresponding to different test concentrations, the Minimum Inhibitory Concentration (MIC) of the three compounds of LZDT1, rifampicin and TCA1 of the invention on H37Rv strain is calculated90) The results are shown in Table 1. The MIC of the LZDT1 compound to H37Rv strain is much higher than the MIC of TCA1 to H37Rv strainMIC of the strain.
TABLE 1 minimum inhibitory concentrations of the compounds LZDT1, rifampicin and TCA1 of the invention against H37Rv strain
And (4) conclusion: the experimental result shows that the antibacterial activity of the compound LZDT1 of the invention on the H37Rv strain is higher, and the antibacterial activity of the compound LZDT1 on the H37Rv strain is equivalent to that of a first-line antituberculosis drug rifampicin.
Experimental example 3 detection of fungicidal Activity of the Compound of the present invention against Mycobacterium tuberculosis H37Rv
Experiment: preparing a 7H10-ADC culture medium plate, treating the mycobacterium tuberculosis for 7 days by using a compound with a concentration corresponding to MIC, coating 100 mu l of bacterial liquid on the 7H10-ADC plate, culturing for 4 weeks at 37 ℃, counting CFU (circulating fluid infusion) and taking the compound concentration with the CFU reduced by 99% as the minimum bactericidal concentration. And determining the influence of the compound on the growth curve of the strain according to the bacteriostatic concentration and bactericidal concentration of the compound.
Mycobacterium tuberculosis with OD value of 0.1 is added into a 15mL centrifuge tube, then 5 mu g/mL compound is added to the centrifuge tube to be cultured in a shaker at 37 ℃ and 80rpm, bacterial liquid is taken and coated on a 7H10-ADC plate after 0 day, 3 days, 7 days, 14 days and 21 days of culture respectively, the bacterial liquid is cultured for 4 weeks at 37 ℃ to count CFU, and a sterilization curve is drawn. While 2. mu.g/mL rifampicin and 5. mu.g/mL TCA1 and DMSO were used as controls.
As a result: LZDT1 was treated at a concentration of 5. mu.g/mL, and after 3 days CFU began to decrease, and after 21 days the bacterial concentration decreased by 4 logs. The results of the experiment are shown in FIG. 4. FIG. 4 is a graph showing the bactericidal activity of the compounds LZDT1 and TCA1 against M.tuberculosis H37 Rv.
And (4) conclusion: the experimental result shows that the bactericidal effect of the compound LZDT1 of the invention is better than that of TCA1 in the first 7 days, and the bactericidal effect of the compound LZDT1 of the invention is equivalent to that of positive control rifampin (rifamycin) in 2 mu g/mL.
Experimental example 4 toxicity test of the Compound of the present invention against human hepatoma cell HepG2
Experiment: expanding and culturing HepG2 cells, and when the cells grow to a logarithmic phase,adjusting the cell density to 3X 104cells/mL, seeded into 96-well cell culture plates, 100. mu.L per well, 3 replicates per concentration gradient. TCA1 and LZDT1 were applied to cells using complete medium at a total of 8 concentration gradients of 0, 1.5625, 3.125, 6.25, 12.5, 25, 50, 100. mu.g/mL, respectively. After 24h, 48h and 72h of incubation, 10. mu.L of CCK-8 was added to each well at 37 ℃ with 5% CO2Culturing in an incubator for 2h in a dark place. OD values at the same time point were measured at a wavelength of 492nm using a microplate reader, and the influence of cell proliferation was analyzed using the measured OD values.
Inhibition (%) - (control OD value-experimental OD value)/(control OD value-blank OD value) × 100%
As a result: TCA1 and intermediate Inhibitory Concentration (IC) of LZDT1 compound in human hepatoma cell HepG2 proliferation for 72h50) 62.57 μ g/mL and 82.78 μ g/mL, respectively. At 24h and 48h, the proliferation inhibition of TCA1 and the compound LZDT1 of the invention on human hepatoma cell HepG2 is not more than 50%. The experimental results are shown in table 2, fig. 5 and fig. 6. FIG. 5 is a toxicity curve of the compound LZDT1 of the present invention on human hepatoma cell HepG 2. FIG. 6 is the toxicity curve of TCA1 on human hepatoma cell HepG 2.
TABLE 2 Medium concentration of the inventive compounds LZDT1 and TCA1 for inhibiting proliferation of human hepatoma cell HepG2
And (4) conclusion: the toxicity of the compound LZDT1 to human hepatoma cells HepG2 is lower than that of TCA1, which shows that the toxicity of the compound LZDT1 to cells is reduced after the compound is modified to TCA 1.
Based on the above description of the summary of the invention, a person skilled in the art can apply the invention in its entirety, and all changes that are the same principle or similar are to be considered as included in the scope of the invention.
Claims (6)
1. Benzothiazole compounds of the following general formula (I) or pharmaceutically acceptable salts thereof,
wherein R1 is:
2. benzothiazole compounds of general formula (I) according to claim 1, or their pharmaceutically acceptable salts, which are the following compounds or their pharmaceutically acceptable salts:
3. a process for preparing benzothiazole compounds of general formula (I) according to claim 1, comprising the steps of:
(1) synthesis of Compound B: adding cyanoacetic acid, the R1 substituted methyl carbamate compound A and acetic anhydride into a reaction vessel, heating in an oil bath, carrying out Ar protection, after full reaction, pouring the reaction solution into ice water, extracting with ethyl acetate, washing with a saturated sodium bicarbonate solution once, drying and concentrating to obtain a compound B;
(2) synthesis of Compound C: adding the compound B, 2, 5-dihydroxy-1, 4-dithiane, methanol and morpholine into a reaction vessel, carrying out oil bath for 2 hours to completely react, directly passing through a column, and eluting to obtain a compound C;
(3) synthesis of Compound (I): adding 2-formic acid benzothiazole and a compound C into a reaction container, adding pyridine, stirring for dissolving, dropwise adding phosphorus oxychloride, after stirring fully, adding dichloromethane and water into the system, quickly stirring to separate an organic layer, washing with a saturated sodium bicarbonate solution once, directly passing through a short silica gel column, and eluting to obtain a compound (I); or adding 2-formic acid benzothiazole and dichloromethane into a reaction container, cooling to 0 ℃, dropwise adding oxalyl chloride, reacting for several hours, adding pyridine into the system, stirring for several minutes, adding a compound C, reacting for several hours, adding methanol into the system to quench the reaction, washing once with a saturated sodium bicarbonate solution, directly mixing the sample with the solution, passing through a silica gel column, and eluting to obtain a compound (I);
wherein R1 is as defined in claim 1.
4. Use of the benzothiazole compounds of general formula (I) according to claim 1 or their pharmaceutically acceptable salts for the preparation of antitubercular agents.
5. Use of a benzothiazole compound of general formula (I) according to claim 1 or its pharmaceutically acceptable salts for the preparation of an anti-tubercular drug as DprE1 inhibitor.
6. Use of the benzothiazole compounds of general formula (I) according to claim 1 or their pharmaceutically acceptable salts for the preparation of antitubercular agents as H37Rv inhibitors.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911079527.7A CN112778297B (en) | 2019-11-07 | 2019-11-07 | Benzothiazole compound and preparation method and application thereof |
| PCT/CN2020/125774 WO2021088753A1 (en) | 2019-11-07 | 2020-11-02 | Benzothiazole compounds, and preparation method therefor and use thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911079527.7A CN112778297B (en) | 2019-11-07 | 2019-11-07 | Benzothiazole compound and preparation method and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN112778297A CN112778297A (en) | 2021-05-11 |
| CN112778297B true CN112778297B (en) | 2022-05-20 |
Family
ID=75747623
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201911079527.7A Active CN112778297B (en) | 2019-11-07 | 2019-11-07 | Benzothiazole compound and preparation method and application thereof |
Country Status (2)
| Country | Link |
|---|---|
| CN (1) | CN112778297B (en) |
| WO (1) | WO2021088753A1 (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116808028B (en) * | 2023-08-25 | 2023-11-14 | 深圳市第三人民医院(深圳市肝病研究所) | Application of benzothiazole derivative compound as antituberculosis compound |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20120095021A1 (en) * | 2010-09-14 | 2012-04-19 | Council Of Scientific And Industrial Research | Synthsis of new benzothiazole derivatives as potential anti-tubercular agents |
| US20130072494A1 (en) * | 2009-02-06 | 2013-03-21 | Elan Pharmaceuticals, Inc. | Inhibitors of jun n-terminal kinase |
| CN105473578A (en) * | 2013-05-24 | 2016-04-06 | 加州生物医学研究所 | Compounds for treatment of drug resistant and persistent tuberculosis |
| CN109988129A (en) * | 2019-04-29 | 2019-07-09 | 中国科学院微生物研究所 | Compound and its preparing the application in antituberculotic |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1986637A1 (en) * | 2006-02-13 | 2008-11-05 | Laboratoires Serono SA | Sulfonamide derivatives for the treatment of bacterial infections |
| WO2009158118A2 (en) * | 2008-05-30 | 2009-12-30 | University Of Notre Dame Du Lac | Anti-bacterial agents from benzo[d]heterocyclic scaffolds for prevention and treatment of multidrug resistant bacteria |
-
2019
- 2019-11-07 CN CN201911079527.7A patent/CN112778297B/en active Active
-
2020
- 2020-11-02 WO PCT/CN2020/125774 patent/WO2021088753A1/en active Application Filing
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20130072494A1 (en) * | 2009-02-06 | 2013-03-21 | Elan Pharmaceuticals, Inc. | Inhibitors of jun n-terminal kinase |
| US20120095021A1 (en) * | 2010-09-14 | 2012-04-19 | Council Of Scientific And Industrial Research | Synthsis of new benzothiazole derivatives as potential anti-tubercular agents |
| CN105473578A (en) * | 2013-05-24 | 2016-04-06 | 加州生物医学研究所 | Compounds for treatment of drug resistant and persistent tuberculosis |
| CN109988129A (en) * | 2019-04-29 | 2019-07-09 | 中国科学院微生物研究所 | Compound and its preparing the application in antituberculotic |
Non-Patent Citations (3)
| Title |
|---|
| Discovery of Pyrazolopyridones as a Novel Class of Noncovalent DprE1 Inhibitor with Potent Anti-Mycobacterial Activity;Manoranjan Panda et al.;《J.Med.Chem.》;20140512;第57卷;第4761-4771页 * |
| Identification of Morpholino Thiophenes as Novel Mycobacterium tuberculosis Inhibitors, Targeting QcrB;Laura A. T. Cleghorn et al.;《J.Med.Chem.》;20180626;第61卷;第6592-6608页 * |
| 苯并噻唑衍生物的生物活性及其构效关系研究进展;武祥龙;《化学与生物工程》;20141231;第31卷(第5期);第1-6、9页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| WO2021088753A1 (en) | 2021-05-14 |
| CN112778297A (en) | 2021-05-11 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| EA035816B1 (en) | Prodrugs of a jak inhibitor compound for treatment of gastrointestinal inflammatory disease | |
| EP1438298B1 (en) | Piperazine derivatives with ccr1 receptor antagonist activity | |
| Thanna et al. | Synthesis and evaluation of new 2-aminothiophenes against Mycobacterium tuberculosis | |
| CN108135887A (en) | Oxazolidinone compounds and its application method as antiseptic | |
| CN106866571A (en) | Heterocyclic urea compound and its pharmaceutical composition and application | |
| Almansour et al. | A solvent free, four-component synthesis and 1, 3-dipolar cycloaddition of 4 (H)-pyrans with nitrile oxides: Synthesis and discovery of antimycobacterial activity of enantiomerically pure 1, 2, 4-oxadiazoles | |
| BR112018004863B1 (en) | COMPOUNDS USED FOR THE PREVENTION OR THERAPEUTIC APPLICATION OF HYPERURICEMIA OR GOUT | |
| CN112778297B (en) | Benzothiazole compound and preparation method and application thereof | |
| Uria-Nickelsen et al. | Novel topoisomerase inhibitors: microbiological characterisation and in vivo efficacy of pyrimidines | |
| Donehower et al. | New developments on the mechanisms of action of antineoplastic drugs | |
| CN111249283A (en) | Pyrimidine derivatives having anticancer effect | |
| EP3371172A1 (en) | Tetrahydroindazoles and medical uses thereof | |
| TWI790024B (en) | Triazolo pyrazine compound and application thereof | |
| WO2011060976A1 (en) | Tryptamine-derived compounds as antibacterial agents | |
| CN110981888B (en) | N-aryl dithiopyrryl ketonuria and amino ester derivatives, preparation and application thereof | |
| CN105163733B (en) | Furan ketone compound as kinase inhibitor | |
| WO2018170664A1 (en) | Oxazolidinone compounds and methods of use thereof as antibacterial agents | |
| KR102198101B1 (en) | Composition for antibiotics against staphylococcus aureus and mycobacterium | |
| Murugesan et al. | 2-Mercapto-quinazolinones as inhibitors of NDH-2 and Mycobacterium tuberculosis: Structure-activity relationships, mechanism of action and ADME characterization. | |
| TW201522316A (en) | Shikimate pathway inhibitors and the use thereof | |
| JPS6252757B2 (en) | ||
| RU2815045C1 (en) | (e)-5-amino-2-oxo-1-((e)-2-oxo-4-phenylbut-3-en-1-ylidene)-1,2,6,7,8,9-hexahydrobenzo[4,5]thieno[3,2-e]pyrrolo[1,2-a]pyrimidine-3-carboxamide, having anticancer activity in pulmonary melanoma therapy | |
| RU2675240C1 (en) | Pyranoindoles with anti-tuberculosis activity | |
| CN108689958A (en) | A kind of indoleamine 2 containing diazanyl, 3- is bis- to add oxidase inhibitor | |
| JPS6122090A (en) | Amino-substituted phenothiazine type rifamycin and its use of drug |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |