CN113332277B - Application of diketopyrrolopyrrole compound in preparation of antifungal drugs - Google Patents

Abstract

The invention discloses a new application of a compound shown as a formula I. The new application is the application of the compound shown in the formula I or the pharmaceutically acceptable salt thereof in preparing products for resisting fungi, particularly Candida albicans infection. According to the invention, the compound shown in the formula I is subjected to anti-Candida albicans activity determination by a microplate double dilution method, and the result shows that the compound shown in the formula I has better bacteriostatic activity on Candida albicans, and is expected to develop a new medicine for treating Candida albicans infection.

Description

Application of diketopyrrolopyrrole compound in preparation of antifungal drugs

Technical Field

The invention belongs to the field of medicines, and particularly relates to an application of a diketopyrrolone compound in preparation of antifungal medicines.

Background

Along with the spread of chronic diseases such as hypertension, hyperlipidemia, diabetes and the like, the increase of the number of patients suffering from cancer radiotherapy, chemotherapy and organ transplantation, the wide use of immunosuppressive agents, the decrease of the immunocompetence of human bodies caused by the global epidemic of AIDS and the like, and the incidence rate of fungal infection is obviously increased. Currently, antifungal drugs used clinically mainly include polyenes (polyenes), allylamines (allyalmines), flucytosines (Flucytosine), and other drugs, but the drug resistance of pathogenic fungi is becoming more severe, and the required dosage has to be gradually increased, so that the research and development of new drugs for treating fungal infection diseases are imperative.

Candida is the most common pathogenic bacterium of fungi, and is also called candida. It often parasitizes on skin, oral cavity, vagina and intestinal mucosa of human, and is liable to cause candida infectious diseases when the immune function of organism is low or the micro-ecological environment of normal parasitizing part is disordered. Clinically common Candida species include Candida albicans (Candida albicans), candida glabrata, and Candida guilliermondii. Among them, candida albicans is a pathogenic fungus widely spread in human, often causing acute, subacute or chronic infection, and is one of the most important pathogens of hospital acquired infection at present. It has been found that candida albicans does not normally cause diseases on mucosal surfaces of healthy persons, such as oral cavity, intestinal tract, etc., but causes serious systemic infection in patients with compromised or suppressed immune systems, such as chemotherapy patients, organ transplant patients or aids patients, with a mortality rate of up to 40%.

Therefore, the development of a novel safe and effective anti-candida albicans infection drug has important practical significance.

Disclosure of Invention

One object of the present invention is to provide a novel use of 1- [4- (difluoromethoxy) phenyl ] pyrrole-2,5-dione or a pharmaceutically acceptable salt thereof.

The 1- [4- (difluoromethoxy) phenyl ] pyrrole-2,5-dione has a CAS No.68255-65-2, and the structural formula is shown as the formula I:

the novel application of the compound shown in the formula I or the pharmaceutically acceptable salt thereof provided by the invention is at least one of the following 1) to 7):

1) The application in preparing fungus bacteriostat;

2) The use thereof for the preparation of a product inhibiting the activity of fungi;

3) The application in preparing products for resisting fungal infection;

4) The application in preparing products for preventing and/or treating diseases caused by fungal infection;

5) Use in inhibiting fungal activity;

6) Use against fungal infections;

7) The application in preventing and/or treating diseases caused by fungal infection.

Another object of the present invention is to provide a fungal bacteriostatic agent.

The active ingredient of the fungus bacteriostatic agent provided by the invention is a compound shown in a formula I or a pharmaceutically acceptable salt thereof.

It is yet another object of the present invention to provide a product.

The active ingredient of the product provided by the invention is a compound shown in a formula I or pharmaceutically acceptable salt thereof;

the product has at least one of the following effects:

a) Inhibiting fungal activity;

b) Resisting fungal infection;

c) Preventing and/or treating diseases caused by fungi.

The product of the invention can be specifically a medicine.

When necessary, one or more pharmaceutically acceptable carriers can be added into the medicine; the carrier includes diluent, excipient, filler, binder, wetting agent, disintegrating agent, absorption enhancer, surfactant, adsorption carrier, lubricant, etc. which are conventional in the pharmaceutical field.

The above medicine can be made into various forms such as injection, tablet, powder, granule, capsule, oral liquid, paste, cream, etc.; the medicaments in various dosage forms can be prepared according to the conventional method in the pharmaceutical field.

The above medicine can be introduced into body such as muscle, intradermal, subcutaneous, intravenous, and mucosal tissue by injection, spray, nasal drop, eye drop, penetration, absorption, physical or chemical mediated method; or mixed or coated with other materials and introduced into body.

The invention also provides a method for inhibiting the activity of fungi.

The method for inhibiting the activity of the fungi comprises the following steps: a substance comprising a compound represented by the formula I or a pharmaceutically acceptable salt thereof as an active ingredient is used for inhibiting the activity of fungi.

The minimum use concentration of the compound shown in the formula I or the pharmaceutically acceptable salt thereof in the substances taking the compound shown in the formula I or the pharmaceutically acceptable salt thereof as the active ingredient is not lower than the Minimum Inhibitory Concentration (MIC) of fungi inhibited by the compound.

The method is a non-disease diagnostic treatment method. For example, as a positive control in screening for drugs sensitive to fungi (e.g., candida albicans).

In the invention, the fungus is specifically candida albicans. The Candida albicans includes Candida albicans sensitive strains (e.g., candida albicans SC 5314), and Candida albicans resistant strains (e.g., C.albicans # 17, C.albicans g 5).

According to the invention, the compound shown in the formula I is subjected to anti-Candida albicans activity determination by adopting a microplate double dilution method, and the result shows that the compound shown in the formula I has better bacteriostatic activity on Candida albicans, and is expected to develop a new medicine for treating Candida albicans infection.

Detailed Description

The present invention will be further illustrated with reference to the following specific examples, but the present invention is not limited to the following examples. The experimental procedures in the following examples are conventional unless otherwise specified. The test materials used in the following examples were purchased from a conventional biochemical reagent store unless otherwise specified. The quantitative tests in the following examples, all set up three replicates and the results averaged.

The following examples used 1- [4- (difluoromethoxy) phenyl ] pyrrole-2,5-dione of formula I, CAS No.68255-65-2, available from Tao Shu Biotech, inc., cat # in Shanghai: f1065-0202;

or according to the method described in patent DE 3144526C 2.

EXAMPLE 1Activity of Compounds of formula I against Candida albicans

RPMI Media 1640, gibco, USA. Fluconazole, itraconazole and amphotericin B Sigma-Aldrich, USA.

1) Preparation of bacterial liquid

Candida albicans sensitive (Candida albicans SC 5314), candida albicans resistant (C.albicans 17#, C.albicans g 5) (Gong Jiao, wu Ju, ikeh Melanie,tao Li, zhang Yulong, bing Jian, nobile Clarisa J., huang Guanghua.Antifungal 1Activity of Mammali Serum amyloid A1 againt Candida albicans and Chemotherpy.64 (1): e 01975-19) prepared at (2-5). Times.10 ⁵ cells/mL of culture.

2) Preparation of test Compounds solutions of formula I

Compound I is prepared into 4mg/mL mother solution by using sterile DMSO as a solvent, and then diluted by the sterile DMSO sequentially to obtain diluted solutions with the concentrations of 2mg/mL, 1mg/mL, 500. Mu.g/mL, 250. Mu.g/mL, 125. Mu.g/mL, 62.5. Mu.g/mL, 31.25. Mu.g/mL, 15.625. Mu.g/mL, 7.81. Mu.g/mL, 3.91. Mu.g/mL, 1.95. Mu.g/mL and 0.98. Mu.g/mL.

The positive control drugs (fluconazole, itraconazole and amphotericin B) are sequentially prepared into mother liquor of 320 mu g/mL by taking sterile DMSO as a solvent, and then the mother liquor is sequentially diluted by using the sterile DMSO to obtain dilution solutions with the concentrations of 160 mu g/mL, 80 mu g/mL, 40 mu g/mL, 20 mu g/mL, 10 mu g/mL, 5 mu g/mL and 2.5 mu g/mL, and the dilution solutions are used as the positive control drugs when the Candida albicans is detected.

3) Determining the minimum inhibitory concentration of the compound of formula I for inhibiting Candida albicans

a. A96-well cell culture plate was prepared, and 40. Mu.L of RPMI Media 1640 medium was added to each well.

b. Taking a 96-well cell culture plate which completes the step a, and grouping the 96-well cell culture plate as follows:

positive control group (7 wells): respectively adding 2 mu L of 7-dilution positive control drug diluent prepared in the step 2);

experimental groups (7 wells per compound): adding 2 mu L of the compound diluent prepared in the step 2) respectively;

negative control group (7 wells): add 2. Mu.L of sterile DMSO each.

c. Taking a 96-well cell culture plate which finishes the step b, adding 40 mu L of the bacterial liquid obtained in the step 1) into each well, culturing at 35 ℃ for 24 hours, and observing the growth condition of candida albicans in each well: if the wells are turbid, the corresponding concentrations of the compound are indicated to have no anti-Candida albicans activity; if the wells are clear, the corresponding concentrations of the compounds are indicated to have anti-Candida albicans activity. For each compound, the final concentration of the compound (concentration of the compound in the added dilution/40) corresponding to the well in which the growth of candida albicans was completely inhibited was the minimum inhibitory concentration, MIC value, of the compound against candida albicans. The results are shown in Table 1

TABLE 1 test results of the anti-Candida albicans activity of the compound of formula I (MIC value, μ g/ml)

As shown in Table 1, the compound 1- [4- (difluoromethoxy) phenyl ] pyrrole-2,5-dione represented by formula I provided by the invention has antifungal effect.

Claims (1)

1. Use of a compound of formula I or a pharmaceutically acceptable salt thereof in at least one of:

(formula I)

1) The application in preparing fungus bacteriostat;

2) The use thereof for the preparation of a product inhibiting the activity of fungi;

3) The application in preparing products for resisting fungal infection;

4) The application in preparing products for preventing and/or treating diseases caused by fungal infection;

the fungus is candida albicans drug-resistant strain;

the candida albicans drug-resistant strain isCandida albicans17 orCandida albicans g5；

The product is a medicine.