Use of benzimidazole compounds for the preparation of a medicament for the prevention or treatment of HPV infection

Abstract

The application discloses a kind of benzimidazole compounds with the following structure, its pharmaceutically acceptable salt and their pharmaceutical composition in the application of preparation prevention and / or treatment HPV infection drug, wherein the inhibition EC 50 Value of compound for high-risk HPV16, HPV18 virus strain reaches micromolar concentration level, EC 50 Value is less than 10 μM, even less than 5 μM, with good anti-HPV infection activity, can be used as primary or auxiliary therapeutic drug, for high-risk, suspected high-risk and low-risk HPV infection, especially high-risk HPV infection;In addition, it also has good safety.

Description

Technical Field

[0001] This invention relates to the use of a benzimidazole compound in the preparation of a drug for the prevention or treatment of HPV infection, and more particularly to the use of a benzimidazole compound in the preparation of a drug for the prevention or treatment of high-risk HPV infection. Background Technology

[0002] HPV (human papillomavirus) is a site-specific DNA virus that replicates and multiplies on the surface of epithelial cells and mucous membranes. Currently, more than 200 subtypes of HPV have been identified, with dozens involved in human genital tract infections. Based on their biological characteristics and pathogenicity, HPV can be divided into low-risk and high-risk types. The vast majority of high-risk HPV types are associated with cervical cancer, primarily HPV16 and HPV18.

[0003] Currently, common clinical interventions for clearing HPV include surgical treatment, physical therapy, and drug therapy. Drug therapy is suitable for cervical HPV infections without abnormal pathology. A wide variety of drugs are used clinically, broadly categorized by their mechanisms of action as immunomodulators or enhancers, including interferon, ligand-receptor drugs such as rulintabubal, and gene therapy drugs such as rebvoigen. Due to the large number of drugs available, there is currently no specific cure or universally accepted treatment, and a systematic theoretical and methodological framework has not been established, leading to considerable confusion in clinical treatment. Therefore, exploring more effective drugs for treating HPV infection is an urgent clinical need. Summary of the Invention

[0004] Purpose of the invention: The present invention aims to provide the use of benzimidazole compounds, pharmaceutically acceptable salts thereof, and pharmaceutical compositions thereof in the preparation of medicaments for the prevention or treatment of HPV infection.

[0005] Technical solution: The benzimidazole compounds having the following structure described in this invention are used in the preparation of drugs for the prevention and / or treatment of HPV infection.

[0006]

[0007] in:

[0008] X is selected from O, NR5, NR6, S, C (=O), CR5R6, CR5R6-CR5R6, and R5 and R6 are selected from H.

[0009] C1-C4 alkyl, halogen; X is more preferably CH2, C(O), CHF, CFF, NH, S;

[0010] R1 and R2 are selected from H, C1-C4 alkyl, cyano, C1-C4 alkylcyano, C1-C4 alkoxy, and 6-10 aryl; R1 is more preferably H, methyl, isopropyl, or cyano; R2 is more preferably H, methyl, or isopropyl.

[0011] R3 and R4 are selected from H, C1-C4 alkyl, cyano, or R3 and R4 form a 3-6 membered cycloalkyl group with the C group to which they are attached; R3 is more preferably H or methyl, R4 is more preferably H or methyl, or R3 and R4 form a cyclopropyl group with the C group to which they are attached.

[0012] Preferably, the drug is a drug for the prevention and / or treatment of HPV infection selected from any of the following:

[0013] HPV16, HPV18, HPV31, HPV33, HPV35, HPV39, HPV45, HPV51, HPV52, HPV56, HPV58, HPV59, HPV66, HPV68, HPV26, HPV53, HPV70, HPV73, HP V82, HPV6, HPV11, HPV34, HPV40, HPV42, HPV43, HPV44, HPV54, HPV55, HPV57, HPV61, HPV67, HPV69, HPV71, HPV72, HPV81, HPV83, HPV84.

[0014] Benzimidazole compounds are known to possess anti-HCV activity and are used to treat hepatitis C or hepatitis C-related conditions. This invention discovers that specially substituted benzimidazole compounds have an inhibitory effect on HPV and can be formulated as drugs for treating human papillomavirus infection. These drugs can be used as primary or adjunctive therapies to treat high-risk (HPV16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, 68), suspected high-risk (HPV26, 53, 70, 73, 82), and low-risk (HPV6, 11, 34, 40, 42, 43, 44, 54, 55, 57, 61, 67, 69, 71, 72, 81, 83, 84) HPV infections, particularly high-risk HPV16 and HPV18 infections.

[0015] Preferably, the structure of the compound is as follows:

[0016] X is CR5R6, where R5 and R6 are selected from H, C1-C4 alkyl groups, and halogens;

[0017] R1 and R2 are selected from H, C1-C4 alkyl, cyano, C1-C4 alkylcyano, and C1-C4 alkoxy.

[0018] R3 and R4 are selected from H, C1-C4 alkyl, cyano, or R3 and R4 can form a 3-6 membered cycloalkyl group with the C group to which they are attached.

[0019] Further preferably, the structure of the compound includes:

[0020] R5 and R6 are halogens;

[0021] R1 and R2 are selected from H, C1-C4 alkyl, cyano, and C1-C4 alkoxy.

[0022] R3 and R4 are selected from H, C1-C4 alkyl, cyano, or R3 and R4 forming a 3-6 membered cycloalkyl group with the C group to which they are attached.

[0023] Further preferably, the structure of the compound includes:

[0024] R5 and R6 are halogens;

[0025] R1 and R2 are selected from H and C1-C4 alkyl groups;

[0026] R3 and R4 are selected from H, C1-C4 alkyl, cyano, or R3 and R4 can form a 3-6 membered cycloalkyl group with the C group to which they are attached.

[0027] More preferably, the structure of the compound includes:

[0028] R1 and R2 are selected from H, methyl, ethyl, propyl, isopropyl, and tert-butyl.

[0029] Furthermore, in a more preferred embodiment, the structure of the compound includes:

[0030] R1 and R2 are selected from H, methyl, ethyl, and isopropyl.

[0031] Specifically, the compound has the following structure:

[0032]

[0033] The pharmaceutically acceptable salts of the benzimidazole compounds described in this invention are also used in the preparation of drugs for the prevention and / or treatment of HPV infection, particularly high-risk HPV16 and HPV18 infections. The pharmaceutically acceptable salt is a salt formed by the benzimidazole compound and an acid selected from any of the following:

[0034] Hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, carbonic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, citric acid, malic acid, tartaric acid, lactic acid, pyruvic acid, acetic acid, maleic acid, succinic acid, fumaric acid, salicylic acid, phenylacetic acid, mandelic acid, ferulic acid.

[0035] The pharmaceutical compositions of the benzimidazole compounds or their pharmaceutically acceptable salts as active ingredients described in this invention are also used in the preparation of drugs for the prevention and / or treatment of HPV infection, especially high-risk HPV16 and HPV18 infection.

[0036] The pharmaceutical composition of this invention is administered orally in a solid form, and the dosage form can be tablets, capsules, granules, pills, powders, etc. Suitable excipients include fillers, such as starches, sugars, celluloses, inorganic salts, etc.; binders, such as starch, sodium carboxymethyl cellulose, hydroxypropyl cellulose, methyl cellulose, ethyl cellulose, hydroxypropyl methyl cellulose, povidone, etc.; disintegrants, such as dry starch, sodium carboxymethyl starch, low-substituted hydroxypropyl cellulose, croscarmellose, croscarmellose sodium; and lubricants, such as magnesium stearate, micronized silica gel, talc, etc.

[0037] Beneficial effects: Compared with the prior art, the present invention has the following significant advantages:

[0038] The compound's inhibitory effect on high-risk HPV16 and HPV18 virus strains in application (EC) 50 The values ​​all reached the micromolar concentration level, EC 50 With a value of less than 10 μM, or even less than 5 μM, it has good anti-HPV infection activity and can be used as a primary or adjunctive treatment for high-risk, suspected high-risk, and low-risk HPV infections, especially high-risk HPV infections; in addition, it has good safety. Detailed Implementation

[0039] The technical solution of the present invention will be further described below with reference to the embodiments.

[0040] Example 1:

[0041] The compounds described in this invention can be obtained by the preparation method disclosed in CN102596936A. Representative compounds used in the aforementioned applications are as follows:

[0042]

[0043]

[0044]

[0045]

[0046]

[0047]

[0048] Example 2: Biological Experiment

[0049] I. Experimental Cells and Virus Strains

[0050] 293FT cells, HPV-16 and HPV-18 VLPs.

[0051] II. Experimental Methods

[0052] 1. Antiviral experiment

[0053] (1) 293FT cells were seeded into microplates at a certain density and cultured overnight in a 5% CO2, 37°C incubator.

[0054] (2) On the second day, 50 μL of the test compound and reference compound (10-fold dilution, 8 concentrations, double replicates) and 50 μL of 100 TCID50 HPV pseudovirus were added to the cell wells. At the same time, cell control (cells, no compound treatment and virus infection) and virus control (cells infected with virus, no compound treatment) were set up.

[0055] The highest test concentration of the test compound is 100,000 nM (0.01, 0.1, 1, 10, 100, 1000, 10000, 10000 nM).

[0056] The highest concentration tested for the reference compound GSI-IX was 10000 nM (0.001, 0.01, 0.1, 1, 10, 100, 1000, 10000 nM).

[0057] (3) The mixture was incubated for 3 days in a 5% CO2, 37°C incubator.

[0058] (4) The antiviral activity of the compounds was expressed as the inhibition rate (%) of the compounds at different concentrations against the cytopathic effect induced by the virus. The inhibition rate of the compounds was analyzed using a nonlinear fitting method with GraphPad Prism to calculate the half-maximal effective concentration (EC50) of the compounds. 50 )value.

[0059] 2. Cytotoxicity test

[0060] (1) 293FT cells were seeded into microplates at a certain density and cultured overnight in a 5% CO2, 37°C incubator.

[0061] (2) On the second day, 50 μL of the diluted test compound and reference compound (10-fold dilution, 8 concentrations, double replicates) and 50 μL of culture medium were added to the cell wells that were not infected with the virus.

[0062] The highest test concentration of the test compound is 100,000 nM (0.01, 0.1, 1, 10, 100, 1000, 10000, 10000 nM).

[0063] The highest concentration tested for the reference compound GSI-IX was 10000 nM (0.001, 0.01, 0.1, 1, 10, 100, 1000, 10000 nM).

[0064] (3) The mixture in 5% CO 2. Incubate at 37℃ for 3 days.

[0065] (4) The cytotoxicity of the compounds was expressed as the viability (%) of 293FT cells at different concentrations. Nonlinear fitting analysis of cell viability was performed using GraphPadPrism to calculate the half-maximal cytotoxic concentration (CMC) of the compounds. 50 )value.

[0066] III. Experimental Results

[0067] Based on the results of biological experiments on some compounds, the following (EC) 50 Value division—A: 0–5 μM, B: 5–10 μM, C: 10–15 μM, D: 15–20 μM):

[0068]

[0069]

[0070] As can be seen from the table above, the compounds of this invention exhibit inhibitory effects on high-risk HPV16 and HPV18 virus strains. 50 The values ​​all reached the micromolar concentration level, and the EC values ​​of some compounds were also high. 50 With a concentration of less than 10 μM, or even less than 5 μM, it exhibits good anti-HPV infection activity, especially against high-risk HPV infection; in addition, it has low cytotoxicity and good safety.

[0071] The pharmaceutical composition of this invention is administered orally in a solid form, and the dosage form can be tablets, capsules, granules, pills, powders, etc. Suitable excipients include fillers, such as starches, sugars, celluloses, inorganic salts, etc.; binders, such as starch, sodium carboxymethyl cellulose, hydroxypropyl cellulose, methyl cellulose, ethyl cellulose, hydroxypropyl methyl cellulose, povidone, etc.; disintegrants, such as dry starch, sodium carboxymethyl starch, low-substituted hydroxypropyl cellulose, croscarmellose, croscarmellose sodium; and lubricants, such as magnesium stearate, micronized silica gel, talc, etc.

Claims

1. The use of a benzimidazole compound having the following structure in the preparation of a drug for the prevention and / or treatment of HPV16 and HPV18 infections. ， in: The definitions of X, R1, R2, R3, and R4 satisfy any of the following conditions: (1) X is C (=O), R1 is methyl, R2 is methyl, and R3 and R4 form cyclopropyl groups with the C they are attached to. (2) X is C (=O), R1 is isopropyl, R2 isopropyl, and R3 and R4 form cyclopropyl groups with the C they are attached to. (3) X is CHF, R1 is methyl, R2 is methyl, and R3 and R4 form cyclopropyl groups with the C they are attached to; (4) X is CHF, R1 is isopropyl, R2 isopropyl, and R3 and R4 form cyclopropyl groups with the C they are attached to. (5) X is CFF, R1 is methyl, R2 is methyl, R3 is H, and R4 is H; (6) X is CFF, R1 is methyl, R2 is methyl, and R3 and R4 form cyclopropyl groups with the C they are attached to. (7) X is CFF, R1 is isopropyl, R2 isopropyl, and R3 and R4 form cyclopropyl groups with the C they are attached to.

2. Use according to claim 1, characterized in that, The compound has the following structure: 。 3. The use of a pharmaceutically acceptable salt of a benzimidazole compound according to any one of claims 1-2 in the preparation of a medicament for the prevention and / or treatment of HPV infection, wherein the HPV infection is HPV16 or HPV18 infection.

4. The use of a pharmaceutical composition having a benzimidazole compound as described in any one of claims 1-2 or a pharmaceutically acceptable salt as described in claim 3 as an active ingredient in the preparation of a medicament for the prevention and / or treatment of HPV infection, wherein the HPV infection is HPV16 or HPV18 infection.

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