Use of 2-chloro-n-(4-isopropylphenyl)acetamide and pharmaceutically acceptable salts thereof as inhibitors of mycobacterium tuberculosis

Abstract

The application discloses a use of 2-chloro-N-(4-isopropylphenyl)acetamide and pharmaceutically acceptable salts thereof as a mycobacterium tuberculosis inhibitor, and belongs to the field of medicines. The 2-chloro-N-(4-isopropylphenyl)acetamide and pharmaceutically acceptable salts thereof are used for preparing a medicine for preventing tuberculosis. An effective component of the mycobacterium tuberculosis inhibitor is 2-chloro-N-(4-isopropylphenyl)acetamide and pharmaceutically acceptable salts thereof. The test result of the 2-chloro-N-(4-isopropylphenyl)acetamide in vitro anti-mycobacterium tuberculosis shows that the 2-chloro-N-(4-isopropylphenyl)acetamide has a better inhibiting effect on the mycobacterium tuberculosis, and the minimum inhibiting concentration is 0.4 mu g / mL.

Description

Technical Field

[0001] This invention relates to pharmaceuticals, and more particularly to the use of 2-chloro-N-(4-isopropylphenyl)acetamide and its pharmaceutically acceptable salts as an inhibitor of Mycobacterium tuberculosis. Background Technology

[0002] 2-Chloro-N-(4-isopropylphenyl)acetamide is an organic synthetic intermediate with a bifunctional active group, combining the reactivity of the chloroacetyl group with the steric hindrance of the isopropylphenyl group. This class of compounds first appeared in the field of herbicide development in the 1970s as a candidate structure for acetolactate synthase (ALS) inhibitors. In recent years, with the rise of the "fragment linking" strategy in medicinal chemistry, its value as a pharmacophore carrier has been rediscovered, showing unique advantages, especially in the optimization of antibacterial and anti-inflammatory lead compounds. Its structural characteristics are significantly correlated with known bioactive molecules such as chlorsulfuron herbicides and arylacetamide analgesics.

[0003] Tuberculosis is a chronic infectious disease caused by Mycobacterium tuberculosis. While the bacteria can invade various organs throughout the body, it primarily affects the lungs, leading to pulmonary tuberculosis. Mycobacterium tuberculosis (M. tuberculosis), commonly known as tubercle bacillus, is the pathogen that causes tuberculosis. Currently, tuberculosis treatment mainly involves chemotherapy. Rifampin, isoniazid, ethambutol, streptomycin, and pyrazinamide are first-line anti-tuberculosis drugs. Surgical treatment can be performed for drug failure or for life-threatening localized lesions. Symptomatic treatment is also important, such as managing fever and preventing hemoptysis. Summary of the Invention

[0004] Purpose of the invention: The purpose of this invention is to provide the use of 2-chloro-N-(4-isopropylphenyl)acetamide and its pharmaceutically acceptable salts; another purpose of this invention is to provide a Mycobacterium tuberculosis inhibitor.

[0005] Technical solution: Use of 2-chloro-N-(4-isopropylphenyl)acetamide and its pharmaceutically acceptable salts for the preparation of drugs for the prevention of tuberculosis.

[0006] Furthermore, its application in the preparation of Mycobacterium tuberculosis inhibitors.

[0007] Furthermore, tuberculosis prevention drugs are introduced into the body through injection, oral administration, spraying, nasal drops, eye drops, penetration, absorption, physical or chemical mediation; or they are introduced into the body or wounds after being mixed or encapsulated with other substances.

[0008] Furthermore, it can be applied using solid, semi-solid, or liquid formulations.

[0009] Furthermore, tuberculosis prevention drugs also include pharmaceutically acceptable carriers; said carriers include one or more of the following pharmaceutically available diluents, excipients, fillers, binders, humectants, disintegrants, absorption enhancers, surfactants, absorbent carriers, and lubricants.

[0010] Furthermore, this includes its cis-trans structure and stereoisomers.

[0011] Furthermore, Mycobacterium tuberculosis includes susceptible bacteria, single-drug resistant bacteria, multidrug resistant bacteria, multidrug resistant bacteria, and extensively drug-resistant Mycobacterium tuberculosis.

[0012] Furthermore, tuberculosis prevention drugs can be formulated into injections, tablets, powders, granules, capsules, oral liquids, ointments, creams, or other suitable dosage forms specified in the pharmacopoeia.

[0013] Furthermore, the tuberculosis prevention drug contains 0.5-99 wt% of 2-chloro-N-(4-isopropylphenyl)acetamide and its pharmaceutically acceptable salts, based on the total weight of the tuberculosis prevention and / or tuberculosis prevention drug.

[0014] A Mycobacterium tuberculosis inhibitor, the active ingredient being 2-chloro-N-(4-isopropylphenyl)acetamide and its pharmaceutically acceptable salt.

[0015] Beneficial effects: The results of the in vitro antibacterial test of 2-chloro-N-(4-isopropylphenyl)acetamide against Mycobacterium tuberculosis showed that 2-chloro-N-(4-isopropylphenyl)acetamide has a good inhibitory effect on Mycobacterium tuberculosis, with a minimum inhibitory concentration of 0.4 μg / mL. Attached Figure Description

[0016] Figure 1 This is a chromatogram of the detection method for 2-chloro-N-(4-isopropylphenyl)acetamide using microdilution.

[0017] Figure 2 This is a schematic diagram of the chemical structure of 2-chloro-N-(4-isopropylphenyl)acetamide. Detailed Implementation

[0018] To make the technical solution of the present invention clearer, the present invention will be further described in detail below with reference to the accompanying drawings and specific embodiments.

[0019] Unless otherwise specified, the experimental methods used in this invention are conventional methods; the materials and reagents used are commercially available unless otherwise specified.

[0020] The 2-chloro-N-(4-isopropylphenyl)acetamide and its pharmaceutically acceptable salts in this invention have the following properties: Figure 2Compounds with the structure shown. Any cis-trans and stereoisomers having this structural feature are contained in 2-chloro-N-(4-isopropylphenyl)acetamide and its pharmaceutically acceptable salts.

[0021] The present invention relates to 2-chloro-N-(4-isopropylphenyl)acetamide and its pharmaceutically acceptable salts, which are used in the preparation of drugs for the prevention and / or treatment of tuberculosis.

[0022] The Mycobacterium tuberculosis in this invention includes susceptible bacteria, single-drug resistant bacteria, multidrug resistant bacteria, multidrug resistant bacteria, and extensively drug resistant Mycobacterium tuberculosis.

[0023] Mycobacterium tuberculosis can be a member of the Mycobacterium tuberculosis complex (MTC).

[0024] The Mycobacterium tuberculosis complex (MTC) comprises Mycobacterium tuberculosis, Mycobacterium africanum, Mycobacterium bovis, Mycobacterium bovis BCG, Mycobacterium canetti, Mycobacterium caprae, Mycobacterium microti, and Mycobacterium pinnipedii. These mycobacteria are the pathogens of tuberculosis in humans and animals. Mycobacterium tuberculosis is the leading cause of tuberculosis in humans.

[0025] The tuberculosis described in this invention is caused by infection with Mycobacterium tuberculosis.

[0026] Mycobacterium tuberculosis may exhibit drug resistance, including single-drug resistance, multidrug resistance, multidrug resistance, and extensive drug resistance.

[0027] In another aspect, the 2-chloro-N-(4-isopropylphenyl)acetamide of the present invention and its pharmaceutically acceptable salts are used to treat diseases caused by Mycobacterium tuberculosis infection, wherein Mycobacterium tuberculosis is selected from the above-mentioned members.

[0028] Diseases caused by Mycobacterium tuberculosis infection include, but are not limited to, pulmonary tuberculosis, tuberculous pleurisy, tuberculous bronchitis, and extrapulmonary tuberculosis, such as tuberculous meningitis, lymph node tuberculosis, intestinal tuberculosis, tuberculous peritonitis, renal tuberculosis, reproductive system tuberculosis, bone and joint tuberculosis, and cutaneous tuberculosis.

[0029] In this invention, "pharmaceutically acceptable" means those compounds (including salts), materials, compositions, and dosage forms that are suitable for contact with human and animal tissues without excessive toxicity, irritation, or other problems or complications, within the limits of reasonable medical judgment, and in proportion to a reasonable benefit / risk ratio.

[0030] Pharmaceutically acceptable salts of this invention include inorganic and organic salts. A preferred class of salts are those formed by the compounds of this invention with acids. Representative pharmaceutically acceptable acid addition salts include, but are not limited to: 4-acetaminobenzoate, acetate, adipic acid salt, alginate, ascorbate, aspartate, besylate, benzoate, hydrogen sulfate, hydrogen tartrate, butyrate, calcium edetate, camphorate, camphor sulfonate, decanoate, and hexanoate. ate), octanoate, cinnamate, citrate, cyclohexanesulfonate, digluconate, 2,5-dihydroxybenzoate, bissuccinate, dodecyl sulfate, edetate (ethylenediaminetetraacetic acid), etopoate (lauryl sulfate), ethane-1,2-disulfonate (ethanedisulfonate), esylate, formate, fumarate, galactate (mucilage), gentianate (2,5-dihydroxybenzoate), gluceptate, gluconeate, etc. Glucuronide, glucuronide, glutamate, glutamate, glyceryl phosphate, glycolate, hexylresorcinol, hippurate, heparin (N,N′-di(dehydroabi)-ethylenediamine), hydrobromide, hydrochloride, hydroiodide, hydroxynaphthylcarboxylate, isobutyrate, lactate, lacturonate, laurate, malate, maleate, malonate, mandelate, mesylate, methyl sulfate, mucilage, naphthalene-1,5-disulfonate (naphthalene disulfonate), naphthalene-2-sulfonate (naphthalene sulfonate), nicotinate Nitrates, oleates, palmitates, p-aminobenzenesulfonates, p-aminosalicylate, pyroxyl naphthalate, pantothenate, pectinates, persulfates, phenylacetates, phenylethyl barbiturates, phosphates, polygalacturonic acid salts, propionates, p-toluenesulfonates, pyroglutamate, pyruvate, salicylates, sebacic acid salts, stearates, hypoacetates, succinates, aminosulfonates, sulfates, tannins, tartrates, theophylline (8-chlorotheophylline), thiocyanates, triethyliodide, undecanoate, undecenoate, and valerate.

[0031] Another preferred class of salts are salts formed by the compounds of the present invention with a base, including but not limited to aluminum, 2-amino-2-(hydroxymethyl)-1,3-propanediol (TRIS, aminobutanetriol), arginine, phenethylbenzylamine (N-benzylphenylethylamine), benzylamine (N,N'-dibenzylethylenediamine), bis-(2-hydroxyethyl)amine, bismuth, calcium, chloroprocaine, choline, crimidazole (1-p-chlorobenzyl-2-pyrrolidine-1′-methylbenzimidazole), cyclohexylamine, diphenylmethylethylenediamine, diethylamine, diethyltriamine, dimethylamine, dimethylethanolamine, dopamine, ethanolamine, ethylenediamine, L-histidine, iron, isoquinoline, p-methylpyridine, lithium, lysine, magnesium, meglumine (N-methylglucosamine), piperazine, piperidine, potassium, procaine, quinine, quinoline, sodium, strontium, tert-butylamine, and zinc.

[0032] In this invention, pharmaceutically acceptable salts are preferably phosphates or ammonium salts.

[0033] The 2-chloro-N-(4-isopropylphenyl)acetamide and its pharmaceutically acceptable salt of the present invention can be prepared by methods well known to those skilled in the art, with no particular limitation on the reaction parameters of each step, and can also be obtained commercially.

[0034] The present invention provides a drug for the prevention and / or treatment of tuberculosis containing a safe and effective amount of 2-chloro-N-(4-isopropylphenyl)acetamide and its pharmaceutically acceptable salts, as well as a pharmaceutically acceptable carrier or excipient. Such carriers include (but are not limited to): saline, buffer solutions, glucose, water, glycerol, ethanol, powders, and combinations thereof. The pharmaceutical formulation should be matched to the route of administration.

[0035] Taking pharmaceutical compositions as an example, the preventive and / or tuberculosis drugs of the present invention can be formulated into injectable forms, for example, prepared by conventional methods using physiological saline or aqueous solutions containing glucose and other excipients. Pharmaceutical compositions such as tablets and capsules can be prepared by conventional methods. Pharmaceutical compositions such as injections, solutions, tablets, and capsules are preferably manufactured under sterile conditions. The preventive and / or tuberculosis drugs of the present invention can also be formulated into powders for nebulized inhalation.

[0036] The tuberculosis prevention and / or tuberculosis medication of the present invention can be introduced into the body, such as muscles, subcutaneous tissue, intradermal tissue, veins, bones, or mucous membranes, by means of injection, oral administration, spray, nasal drops, eye drops, penetration, absorption, or physical or chemical mediation; or it can be introduced into the body or wound after being mixed or encapsulated with other substances.

[0037] Drugs for the prevention and / or treatment of tuberculosis prepared from 2-chloro-N-(4-isopropylphenyl)acetamide and its pharmaceutically acceptable salts can be formulated into various forms, including injections, tablets, powders, granules, capsules, oral liquids, ointments, and creams. All of these dosage forms can be prepared according to methods used in the pharmaceutical field.

[0038] Example

[0039] Mycobacterium tuberculosis H37Rv was the quality control strain for drug susceptibility proficiency testing at the National Tuberculosis Reference Laboratory of the Chinese Center for Disease Control and Prevention in 2024.

[0040] The microdilution method is used, and the specific operating steps are as follows:

[0041] (1) Transplantation culture:

[0042] H37Rv was aseptically transferred to modified LJ medium and cultured at 37°C for 28 days.

[0043] (2) Prepare drug-containing tablets:

[0044] (21) Preparation of compound storage solution: Based on the weight and potency of 2-chloro-N-(4-isopropylphenyl)acetamide, dissolve it in dimethyl sulfoxide (DMSO) to prepare a drug storage solution with a concentration of 10 mg / mL.

[0045] (22) Serial dilution:

[0046] (221) Add 99.5 μL of 7H9 liquid medium containing 10% OADC to the second column of the 96-well plate, add 50 μL to the remaining columns, and add 200 μL of PBS per well to the outermost ring of the 96-well plate to prevent drying.

[0047] (222) Add 0.51 μL of 10 mg / mL 2-chloro-N-(4-isopropylphenyl)acetamide to each well in the second column of a 96-well plate to achieve a maximum working concentration of 51.2 μg / mL. After mixing by pipetting, add 50 μL of the mixture to each well in the second column and mix with 50 μL of liquid culture medium. Repeat this process up to the tenth column, discarding 50 μL of the mixture after each mixing. This process was repeated to achieve compound concentrations of 102.4, 51.2, 25.6, 12.8, and 0.4 μg / mL.

[0048] (223) Set DMSO as a negative control, rifampin as a positive control, and a blank control. Each compound was tested in triplicate. Repeat 3 times.

[0049] (3) Prepare bacterial suspension:

[0050] (31) Scrape colonies from the solid culture medium into a saline solution containing 0.2% Tween and glass beads. Vortex for at least 30 seconds. After the bacterial suspension has settled naturally for 15 minutes, adjust the turbidity to 0.5 McFarland standard using a turbidimeter.

[0051] (32) Transfer 15 μL of bacterial suspension to 7H9 liquid culture medium containing 15 ml of 10% OADC and vortex to mix for 30 seconds. Obtain an inoculum of 1×10⁵ CFU / mL (range 5×10⁴ to 5×10⁵ CFU / mL). Complete the preparation and transfer of bacterial suspension within 30 minutes.

[0052] (4) Inoculation: Add 50 μL of bacterial suspension to each well of the drug-containing microplate and make the final concentration of the compound 51.2, 25.6, 12.8 to 0.2 μg / mL.

[0053] (5) Sealing: Cover all wells with adhesive sealing film to ensure that all wells are completely covered. After capping the 96-well cell culture plate, place it in a transparent plastic sealing bag and heat-seal it to ensure sufficient airtightness.

[0054] (6) Incubation: Incubate at 37℃ for 7-10 days and check its growth. If the growth is not good after 10 days, put it back into the incubator and continue incubating the drug sensitivity plate for 11 days. During the incubation process, a maximum of two plates can be stacked.

[0055] (7) Interpretation of minimum inhibitory concentration (MIC) results: (as shown in Table 1)

[0056] (71) Results can be read visually or using a microplate bottom scanner (do not remove the sealing film). Growth is characterized by turbidity or colony deposition at the bottom of the wells. The MIC value is the lowest compound concentration that significantly inhibits growth.

[0057] (72) Read the blank, negative and positive control wells of reference strain H37Rv. If there is growth of the strain in the blank control well, the result is invalid, indicating that the liquid culture medium is contaminated. If there is no positive growth in the negative control well, the result is invalid, indicating that the DMSO concentration affects the bacterial activity. If positive growth appears in the positive control well outside the quality control concentration range of the quality control strain, the result is invalid, indicating that there is a problem in the operation process.

[0058] PBS PBS PBS PBS PBS PBS PBS PBS PBS PBS PBS PBS PBS 51.2 25.6 12.8 6.4 3.2 1.6 0.8 0.4 0.2 0.1 PBS PBS 51.2 25.6 12.8 6.4 3.2 1.6 0.8 0.4 0.2 0.1 PBS PBS 51.2 25.6 12.8 6.4 3.2 1.6 0.8 0.4 0.2 0.1 PBS PBS Positive blank Negative / / / / / / / PBS PBS Positive blank Negative / / / / / / / PBS PBS Positive blank Negative / / / / / / / PBS PBS PBS PBS PBS PBS PBS PBS PBS PBS PBS PBS PBS

[0059] Drug concentration unit: μg / mL

[0060] The experiment showed that 2-chloro-N-(4-isopropylphenyl)acetamide has a good inhibitory effect on Mycobacterium tuberculosis, with a minimum inhibitory concentration of 0.4.

[0061] The embodiments described above are merely illustrative of several implementations of the present invention, and while the descriptions are specific and detailed, they should not be construed as limiting the scope of the present invention. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these modifications and improvements all fall within the scope of protection of the present invention. Therefore, the scope of protection of this patent should be determined by the appended claims.

Claims

1. The use of 2-chloro-N-(4-isopropylphenyl)acetamide and its pharmaceutically acceptable salts, characterized in that, This is used to prepare drugs for the prevention of tuberculosis caused by Mycobacterium tuberculosis H37Rv.

2. The use of 2-chloro-N-(4-isopropylphenyl)acetamide and its pharmaceutically acceptable salts according to claim 1, characterized in that, Application in the preparation of Mycobacterium tuberculosis H37Rv inhibitors.

3. The use of 2-chloro-N-(4-isopropylphenyl)acetamide and its pharmaceutically acceptable salts according to claim 1, characterized in that, Tuberculosis prevention drugs are introduced into the body through physical or chemical means; or they are introduced into the body or wounds after being mixed or encapsulated with other substances.

4. The use of 2-chloro-N-(4-isopropylphenyl)acetamide and its pharmaceutically acceptable salts according to claim 3, characterized in that, It can be applied using solid, semi-solid, or liquid formulations.

5. The use of 2-chloro-N-(4-isopropylphenyl)acetamide and its pharmaceutically acceptable salts according to claim 1, characterized in that, Tuberculosis prevention drugs also include pharmaceutically acceptable carriers.

6. The use of 2-chloro-N-(4-isopropylphenyl)acetamide and its pharmaceutically acceptable salts according to claim 2, characterized in that, Mycobacterium tuberculosis H37Rv is a susceptible bacterium.

7. The use of 2-chloro-N-(4-isopropylphenyl)acetamide and its pharmaceutically acceptable salts according to claim 1, characterized in that, Tuberculosis prevention drugs can be formulated into injections, tablets, powders, granules, capsules, oral liquids, ointments, creams, or other suitable dosage forms specified in the pharmacopoeia.

8. The use of 2-chloro-N-(4-isopropylphenyl)acetamide and its pharmaceutically acceptable salts according to claim 1, characterized in that, Tuberculosis prevention drugs contain 0.5-99 wt% of 2-chloro-N-(4-isopropylphenyl)acetamide and its pharmaceutically acceptable salts, based on the total weight of the prevention and / or tuberculosis drugs.

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