Application of cepharanthin combined with terbinafine hydrochloride in the preparation of drugs for drug-resistant Candida albicans

Abstract

The application discloses application of Stephania sinica and terbinafine hydrochloride in preparation of a drug for drug-resistant Candida albicans. When Stephania sinica and terbinafine hydrochloride are combined, a significant synergistic effect is shown on drug-resistant Candida albicans, and the bacteriostatic effect on Candida albicans is significant.

Description

Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to application of cepharanthin in combination with terbinafine hydrochloride in preparing a drug-resistant Candida albicans drug. Background Art

[0002] The statements herein merely provide background information related to the present invention and do not necessarily constitute prior art.

[0003] The morbidity and mortality rates of fungal infections continue to rise, and the drug resistance situation is worrying. Common pathogenic fungi include Candida albicans, Aspergillus, etc.

[0004] Candida albicans, also known as Candida albicans, is a widespread fungus in nature. It is an opportunistic pathogen and the most commonly isolated fungus in clinical practice. When the human immune system is weakened or the bacterial flora is imbalanced, Candida albicans can cause infections such as thrush, pneumonia, enteritis, bronchitis, pyelonephritis, meningitis, and endocarditis.

[0005] The treatment of Candida albicans generally includes topical medications, oral medications and intravenous medications. Topical medications include miconazole nitrate suppositories, ketoconazole cream, and ciclopirox cream; oral medications include itraconazole capsules, terbinafine hydrochloride tablets, fluconazole capsules, etc.; intravenous medications include fluconazole sodium chloride injection, fluconazole glucose injection, voriconazole injection, etc.

[0006] In recent years, the incidence of Candida albicans infection has been on the rise, driven by the increasing number of AIDS patients, the widespread use of highly effective broad-spectrum antibiotics and corticosteroids, the advancement of anti-tumor therapy, and the continued implementation of organ transplantation and interventional therapy. Although a variety of antifungal drugs, such as imidazoles, polyenes, and allylamines, are currently available to effectively prevent and treat candidiasis, the widespread use of these drugs has led to an increasing number of drug-resistant Candida albicans isolated. Existing antifungal drugs are ineffective against these resistant strains, necessitating the urgent need for new therapeutic approaches to combat this resistant strain. Summary of the Invention

[0007] In view of the shortcomings of the existing technology, the purpose of the present invention is to provide a use of cepharanthin in combination with terbinafine hydrochloride in the preparation of a drug-resistant Candida albicans drug.

[0008] In order to achieve the above object, the present invention is implemented through the following technical solutions:

[0009] In a first aspect, the present invention provides a use of cepharanthin in combination with terbinafine hydrochloride in the preparation of a drug-resistant Candida albicans drug.

[0010] Stephania japonica was originally used in traditional Chinese medicine. It is the root of Stephania japonica (Thunb.) Miers, a plant of the Menispermaceae family. It was first reported by botanist Bunzo Hayata in 1914 and purified and named stephania in 20 years by Professor Heisaburo Kondo, a pharmacist at the University of Tokyo. The chemical formula of stephania is C 37 H 38 N2O6, white crystals.

[0011] The combination of cepharanthin and the existing antifungal drug terbinafine hydrochloride has a synergistic effect against drug-resistant Candida albicans, which is of great significance.

[0012] In some embodiments, the drug-resistant Candida albicans is a multidrug-resistant Candida albicans that is highly resistant to imidazole, polyene, and allylamine antifungal drugs.

[0013] In some embodiments, the concentration ratio of cepharanthin and terbinafine hydrochloride when used in combination is 2-8:16-32.

[0014] In some embodiments, when cepharanthin is used in combination with terbinafine hydrochloride, the minimum inhibitory concentration of cepharanthin is 2 μg / mL, and the minimum inhibitory concentration of terbinafine hydrochloride is 16 μg / mL.

[0015] In a third aspect, the present invention provides an anti-drug-resistant Candida albicans drug, which includes a pharmaceutical composition of cepharanthin and terbinafine hydrochloride.

[0016] In some embodiments, the drug further includes a pharmaceutically acceptable carrier or excipient, and the carrier or excipient is selected from at least one of a hydrating agent, a thickener, an emulsifier, a preservative, a stabilizer, an ion exchanger, a glidant, an adhesive, a colorant, a flavoring agent, a sweetener, a precipitation inhibitor, a lubricant, a dispersant, a diluent, a flavoring agent, an antioxidant, an isotonic agent, a suspending agent, an emulsification accelerator, a buffer, a disintegrant, a surfactant, an absorbent, a release agent, and a coating agent.

[0017] The beneficial effects achieved by one or more embodiments of the present invention are as follows:

[0018] Experiments have shown that the combined use of cephalothin and terbinafine hydrochloride exhibits significant synergistic effects against drug-resistant Candida albicans and has a significant antibacterial effect against Candida albicans. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] The accompanying drawings, which constitute a part of the present invention, are used to provide a further understanding of the present invention. The exemplary embodiments of the present invention and their descriptions are used to explain the present invention and do not constitute improper limitations on the present invention.

[0020] Figure 1This is a comparison of the hyphae of cepharanthin combined with terbinafine hydrochloride against drug-resistant Candida albicans. The scale bar is 50μm. DETAILED DESCRIPTION

[0021] It should be noted that the following detailed description is illustrative and is intended to provide further explanation of the present invention. Unless otherwise specified, all technical and scientific terms used in the present invention have the same meaning as commonly understood by those skilled in the art to which the present invention belongs.

[0022] The present invention will be further described below with reference to the embodiments.

[0023] Example 1

[0024] Experimental process and methods

[0025] Preparation of drug stock solution

[0026] Dissolve cepharanthin powder in DMSO to a concentration of 20480 μg / mL, store in sterile cylindrical drug susceptibility tubes, label, and store in a -20°C refrigerator until use. Dissolve terbinafine hydrochloride bulk drug powder in anhydrous ethanol to a concentration of 20480 μg / mL, store in sterile cylindrical drug susceptibility tubes, label, and store in a -20°C refrigerator until use.

[0027] Preparation of strains

[0028] Use a disposable inoculation loop to streak the stored drug-resistant Candida albicans strain onto YPD solid medium, incubate at 35°C for 48 hours, and subculture three times to fully activate the strain. Pick a single colony from the above medium and resuspend it in YPD liquid medium. Incubate at 35°C overnight. Use a McFadden turbidimeter tube to adjust the bacterial suspension to 2×10 6 / mL, for future use.

[0029] Drug combination effect determination

[0030] This example uses a sterile 96-well plate and performs standard operations in accordance with the American Clinical and Laboratory Standards Institute CLSI (M27). First, two drug working solutions of different gradient concentrations are added to the wells of the 96-well plate, 50 μL per well (final concentration of terbinafine hydrochloride: 1 μg / mL, 2 μg / mL, 4 μg / mL, 8 μg / mL, 16 μg / mL, 32 μg / mL, 64 μg / mL, 128 μg / mL, 256 μg / mL, 512 μg / mL, final concentration of cepharanthin: 16 μg / mL, 32 μg / mL, 64 μg / mL, 128 μg / mL, 256 μg / mL, 512 μg / mL); then the prepared standard bacterial suspension (final concentration: 10 3Add 100 μL of the bacterial suspension (100 μL / mL) to the wells of a 96-well plate containing the drug solution. Add 100 μL of bacterial suspension and 100 μL of culture medium to the growth control wells. The last row of wells serves as a negative control (blank control) containing only culture medium. Fill the remaining wells of the drug sensitivity plate to 200 μL with culture medium. After the preparation is complete, place the plate in a constant temperature incubator and incubate at 35°C for 24 hours, then observe the results.

[0031] Interpretation of experimental results

[0032] Refer to CLSI standards, compare with the growth control wells during interpretation, and visually interpret the bacterial growth in each well under natural light. MIC is defined as the drug concentration that can inhibit more than 80%.

[0033] Synergy evaluation

[0034] In the laboratory, the fractional inhibitory concentration index (FICI) is used as the basis for judging the effectiveness of the combined drug sensitivity test: FICI = FIC A +FIC B =MIC A Combined use ÷MIC A Single use + MIC B Combined use ÷MIC B For single use. Where: MIC A and MIC B is the MIC value of drugs A and B when used alone; C A and C B The MIC values ​​of drugs A and B when used together. The effectiveness of the combined drug susceptibility test can be determined based on the range of FICI: FICI ≤ 0.5 indicates synergistic effect.

[0035] Table 1 Effect of cepharanthin combined with terbinafine hydrochloride against 6 strains of drug-resistant Candida albicans

[0036]

[0037] A: Terbinafine hydrochloride; B: Cephalaenopsis indole.

[0038] Mycelium inhibition experiment

[0039] The fungal cells were cultured overnight and the bacterial concentration was adjusted to 1×10 5 cells / mL, different concentrations of drugs were added. The first group was the control group, which did not receive drugs; the second group was added with 1 μg / ml terbinafine hydrochloride; the third group was added with 16 μg / ml cepharanthin; the fourth group was added with 1 μg / ml terbinafine hydrochloride + 16 μg / ml cepharanthin.

[0040] After static culture at 37°C for 3 h (1), 6 h (2), and 24 h (3), the mycelial morphology was observed under a microscope, e.g. Figure 1 shown.

[0041] Experimental Conclusion

[0042] The combination of cephalanthrin and terbinafine hydrochloride synergistically inhibits drug-resistant Candida albicans, showing a significant synergistic effect. Furthermore, the combination of cephalanthrin and terbinafine hydrochloride significantly inhibits hyphae, an important virulence factor of drug-resistant Candida albicans.

[0043] The foregoing description is merely a preferred embodiment of the present invention and is not intended to limit the present invention. Those skilled in the art will readily appreciate that various modifications and variations of the present invention are possible. Any modifications, equivalent substitutions, or improvements made within the spirit and principles of the present invention are intended to be within the scope of protection of the present invention.

Claims

1. Use of cepharanthin combined with terbinafine hydrochloride in the preparation of an anti-drug-resistant Candida albicans drug; the drug-resistant Candida albicans is multidrug-resistant Candida albicans; the concentration ratio of cepharanthin to terbinafine hydrochloride in the anti-drug-resistant Candida albicans drug is 2-8:16-32; the drug also includes a pharmaceutically acceptable carrier.

2. The use according to claim 1, characterized in that: When cephalanthrin and terbinafine hydrochloride were used in combination, the minimum inhibitory concentration of cephalanthrin was 2 μg / mL, and the minimum inhibitory concentration of terbinafine hydrochloride was 16 μg / mL.

3. An anti-drug-resistant Candida albicans drug, characterized in that: The composition comprises a composition of cepharanthin and terbinafine hydrochloride, wherein the concentration ratio of the cepharanthin to terbinafine hydrochloride is 2-8:16-32; the medicine further comprises a pharmaceutically acceptable carrier.

4. The anti-drug-resistant Candida albicans drug according to claim 3, characterized in that: The drug further includes a pharmaceutically acceptable carrier, which is selected from at least one of a thickener, an emulsifier, a preservative, a stabilizer, an ion exchanger, a glidant, a binder, a colorant, a flavoring agent, a sweetener, a precipitation inhibitor, a lubricant, a dispersant, a diluent, an antioxidant, an isotonic agent, a suspending agent, a buffer, a disintegrant, a surfactant, an absorbent, a release agent, and a coating agent.

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