A compound and its use in the preparation of antifungal and antitumor drugs

Abstract

The application discloses a compound and application thereof in preparation of antifungal and antitumor drugs, and relates to the technical field of medicines.The application relates to application of a compound in preparation of antifungal and antitumor drugs.The compound can not only effectively inhibit growth of fungi, but also has a significant inhibitory effect on proliferation of human lung cancer cells (PC-9) and breast cancer cells (MCF-7). Therefore, the compound can be used as an ideal candidate for preparation of drugs with antifungal and antitumor activities, and has important development value and clinical application prospect.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical technology, and in particular to a compound and its use in the preparation of antifungal and antitumor drugs. Background Technology

[0002] In recent years, invasive fungal infections (IFI) and malignant tumors have become major public health problems threatening human health. Current mainstream antifungal drugs (such as azoles and polyenes) face an increasingly serious drug resistance crisis. Globally, the rate of fungal drug resistance is rising year by year, and the emergence of cross-resistant strains and multidrug-resistant bacteria has significantly reduced the success rate of clinical treatment, forcing the medical field to urgently explore new antibacterial strategies. At the same time, traditional antitumor drugs (such as chemotherapy drugs and targeted therapies) generally suffer from insufficient selectivity and significant toxic side effects, resulting in limited clinical efficacy. For example, chemotherapy drugs often cannot precisely target tumor tissue due to their toxicity to normal cells, thus limiting treatment effectiveness.

[0003] For patients with malignant tumors complicated by fungal infection, the combined use of antifungal drugs and antitumor drugs faces multiple challenges, mainly in the following aspects: (1) Drug interaction: the two types of drugs may conflict through the CYP450 enzyme metabolism system, affecting efficacy or aggravating toxicity; (2) Pharmacokinetic mismatch: differences in drug absorption, distribution, metabolism and excretion processes may make it difficult to coordinate the treatment window and increase the risk of adverse reactions.

[0004] Recent studies have revealed that fungi (such as Aspergillus) colonizing the tumor microenvironment may promote tumor progression and further exacerbate patients' conditions by regulating immune responses or metabolic pathways. This finding highlights the necessity of developing compounds with both antifungal and antitumor activities.

[0005] To address the aforementioned challenges, multi-target drug design has become a research hotspot. Such drugs aim to simultaneously inhibit fungal growth and tumor cell proliferation, and their advantages include: (1) achieving dual inhibition of fungi and tumors through a single drug, reducing the complexity of treatment regimens; (2) multi-target action can reduce dependence on a single target and delay the development of drug resistance; (3) effectively avoiding drug interaction problems in traditional combination therapy and improving patient prognosis.

[0006] In response to the need for prevention and treatment of the synergistic threat of IFI and malignant tumors, the development of multi-target dual-effect molecules is not only a revolution in clinical treatment models, but also provides a potential solution to the challenges of drug resistance and drug interactions.

[0007] Based on this, the present invention is proposed. Summary of the Invention

[0008] The purpose of this invention is to provide a compound and its application in the preparation of antifungal and antitumor drugs, so as to solve the problems existing in the prior art.

[0009] To achieve the above objectives, the present invention provides the following solution:

[0010] One of the technical solutions of the present invention is to provide the application of a compound in the preparation of antifungal and antitumor drugs, wherein the structure of the compound is shown below:

[0011] .

[0012] Furthermore, the fungi include yeasts and / or filamentous fungi; the tumors include lung cancer and / or breast cancer.

[0013] Furthermore, the yeast includes one or more of Candida glabrata, Cryptococcus neoformans, Candida auris, Candida krusei, Candida albicans, and Candida parapsilosis; the filamentous fungi include one or more of Aspergillus fumigatus, Fusarium, and Mucor.

[0014] The second technical solution of the present invention provides an antifungal and antitumor pharmaceutical composition, the active ingredient of which includes compounds with the following structures:

[0015] .

[0016] Furthermore, the mass content of the compound in the pharmaceutical composition is 0.1-99%.

[0017] Furthermore, the pharmaceutical composition also includes a pharmaceutically acceptable carrier.

[0018] Furthermore, the carrier includes one or more of the following: diluent, excipient, filler, binder, wetting agent, disintegrant, absorption promoter, surfactant, adsorbent carrier, and lubricant.

[0019] Furthermore, the dosage form of the pharmaceutical composition includes injection, tablet, powder, granule, capsule, oral liquid, ointment or cream.

[0020] Furthermore, the fungi include yeasts and / or filamentous fungi; the tumors include lung cancer and / or breast cancer.

[0021] Furthermore, the yeast includes one or more of Candida glabrata, Cryptococcus neoformans, Candida auris, Candida krusei, Candida albicans, and Candida parapsilosis; the filamentous fungi include one or more of Aspergillus fumigatus, Fusarium, and Mucor.

[0022] The present invention discloses the following technical effects:

[0023] This invention relates to the application of a compound in the preparation of antifungal and antitumor drugs. This compound not only effectively inhibits fungal growth but also exhibits significant inhibitory effects on the proliferation of human lung cancer cells (PC-9) and breast cancer cells (MCF-7). Therefore, this compound is an ideal candidate for preparing drugs with both antifungal and antitumor activities, possessing significant developmental value and promising clinical application prospects. Detailed Implementation

[0024] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as a limitation of the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0025] It should be understood that the terminology used in this invention is merely for describing particular embodiments and is not intended to limit the invention. Furthermore, with respect to numerical ranges in this invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Any stated value or intermediate value within a stated range, as well as each smaller range between any other stated value or intermediate value within said range, is also included in this invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.

[0026] Unless otherwise stated, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. While only preferred methods and materials have been described herein, any methods and materials similar or equivalent to those described herein may be used in the implementation or testing of this invention. All references to this specification are incorporated by way of citation to disclose and describe methods and / or materials associated with those references. In the event of any conflict with any incorporated reference, the content of this specification shall prevail.

[0027] Various modifications and variations can be made to the specific embodiments described in this specification without departing from the scope or spirit of the invention, as will be apparent to those skilled in the art. Other embodiments derived from this specification will also be readily apparent to those skilled in the art. This specification and embodiments are merely exemplary.

[0028] The terms “include,” “including,” “have,” “contain,” etc., used in this article are all open-ended terms, meaning that they include but are not limited to.

[0029] It should be noted that any aspects not described in detail in this invention are conventional practices in the field and are not the focus of this invention.

[0030] Example 1

[0031]

[0032] 9-Methoxy-1H-phenanthrene-1-one (1.5 g, 7.14 mmol) was dissolved in a mixture of 20 ml tetrahydrofuran and 2 ml methanol. Triethylamine (1.44 g, 14.28 mmol) was then added, and the reaction mixture was stirred for 10 min. 2-Aminomethylpyridine (1.54 g, 14.28 mmol) was then added, and the reaction was heated to 60 °C and carried out for 24 hours. After the reaction was completed, the mixture was cooled to room temperature, and 5 ml of deionized water was added to quench the reaction. The reaction mixture was extracted with dichloromethane, separated, dried over anhydrous sodium sulfate, and purified by column chromatography to obtain 1.3 g of the title compound as an orange-red solid, with a yield of 63.7%.

[0033] The structure of the prepared product was identified by nuclear magnetic resonance. 1 The following are the H NMR (400 MHz, DMSO-d6) characterizations:

[0034] δ 8.66 (1H), 8.27 (1H), 8.03 (1H), 8.00 (1H), 7.87 (1H), 7.82 (1H), 7.74 (1H), 7.56 (1H), 7.22 (1H), 7.16 (1H), 7.00 (1H), 5.53 (1H), 4.50 (2H).

[0035] Example of effect verification:

[0036] 1. Antifungal activity test

[0037] (1) Test strains: Nine common pathogenic fungal standard strains were selected as test subjects, namely six yeasts: Candida glabrata CBS 138, Cryptococcus neoformans H99S, Candida glabrata 0250100231, Candida krusei 0250302509, Candida albicans SC 5314, and Candida parapsilosis CDC317; and three filamentous fungi: Aspergillus fumigatus Af293, Fusarium 20257823, and Mucor 0250702805.

[0038] (2) Test methods:

[0039] a. Preparation of bacterial suspension: The corresponding strain was thawed and inoculated onto YPD solid medium using an inoculation loop, and grown at 30°C for 24 h to bring the fungus into the exponential growth phase. After two subcultures, single colonies or hyphae of the corresponding strain were picked and placed into a fungal suspension tube containing sterile physiological saline. The concentration of the bacterial suspension was adjusted to 0.5 Mcf using a McFarland turbidimeter.

[0040] b. Preparation of the compound drug solution: The compound drug prepared in Example 1 was first dissolved in DMSO, then diluted with culture medium and Tween 80 to prepare a concentration of 512 µg / mL. -1 The solution for the positive control group was prepared by dissolving the drug in sterile water and then diluting it with culture medium to a concentration of 256 µg / mL. -1 The solution.

[0041] c. Experimental procedure: 100 µl of culture medium was added to each well of a 96-well plate. 100 µl of the prepared drug was added to the first well. After mixing by pipetting, 100 µl was transferred to the second well. This process was repeated serially up to the eleventh well, which served as the negative control group with only culture medium and drug added. Finally, 100 µl of the corresponding bacterial suspension was added to each well except the eleventh well. The twelfth well served as the positive control group with only culture medium and bacteria added. The plates were then incubated for 24 hours.

[0042] This compound effectively inhibited the growth of nine common pathogenic fungal standard strains. The results of the antifungal activity test are shown in Table 1.

[0043] Table 1. Results of antifungal activity tests against standard strains of 9 common pathogenic fungi.

[0044]

[0045] 2. Antitumor activity test

[0046] (1) Experimental tumor cells: Two common tumor cell lines were selected as experimental subjects: human lung cancer cells PC-9 and human breast cancer cells MCF-7. Both cell lines were purchased from the Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences.

[0047] (2) Test methods:

[0048] a. Cell culture medium preparation: Add 10% fetal bovine serum and 1% penicillin antibiotics to 45 mL of RPMI-1640 medium.

[0049] b. Cell preparation: Human lung cancer cells PC-9 and human breast cancer cells MCF-7 in logarithmic growth phase were selected for the experiment. Frozen cells were removed from the -80°C freezer and thawed in a preheated water bath until they reached the appropriate cell density.

[0050] c. Preparation of compound solutions: The compound was added to serum-containing cell culture medium to prepare concentrations of 10, 50, 100, and 150 μg / mL. -1 The solution.

[0051] d. Cell seeding: Add 1 mL of 0.25% trypsin to the cells and incubate at 37°C with 5% CO2 for 5 min to digest and detach the adherent cells. Centrifuge (1000 rpm, 5 min) and count the cells to ensure an appropriate cell concentration.

[0052] e. Cell seeding: Add 100 µL of cell suspension to each well of a 96-well plate, with a cell count of 6000 cells / well. Set up 3 replicates. Also set up a blank control group with only culture medium and no test solution and a cell control group with only cells and culture medium.

[0053] f. Cultivation and Treatment

[0054] Culture conditions: Place the 96-well plate in an incubator at 37°C and 5% CO2 for 24 h to allow the cells to adhere.

[0055] Drug treatment: After the cells adhered, 100 μL of the prepared concentration gradient drug solution was added to each well of the tumor cells, and the cells were cultured for another 72 h.

[0056] Positive control drug: The molecular weight of powdered cisplatin is 300.05 g / mol, and its solubility in water is 1 mg / mL, corresponding to a concentration of 3.33 mM / L. In preparation, 1 mg of the cisplatin powder was weighed and added to 1 mL of pure water. The solution was dissolved by sonication to prepare a stock solution. This stock solution was then diluted with a culture medium to prepare drug solutions with concentrations of 3.0005 µg / mL, 6.001 µg / mL, 12.002 µg / mL, 24.004 µg / mL, and 48.008 µg / mL, respectively.

[0057] g. MTS reagent addition and incubation

[0058] Add MTS reagent: Prepare the reagent at a ratio of MTS:RPMI-1640 medium = 1:9, and add 100 μL of MTS reagent to each well of tumor cells (including the control group) and blank group (protect from light).

[0059] Incubation: Place the 96-well plate back into an incubator at 37°C and 5% CO2 and incubate for 1 h.

[0060] h. Absorbance measurement: After incubation, the absorbance of each well was measured at a wavelength of 490 nm using a microplate reader.

[0061] i. Data analysis: Cell viability was calculated based on absorbance values. Using the absorbance of the control group as a benchmark, the percentage of cell viability in each treatment group was calculated, and then the IC50 was calculated.

[0062] Cell viability (%) = (OD value of drug-treated group - OD value of blank group) / (OD value of control group - OD value of blank group) × 100%.

[0063] This compound can effectively inhibit the proliferation of human lung cancer cells PC-9 and human breast cancer cells MCF-7. The results of the antitumor activity test are shown in Table 2.

[0064] Table 2

[0065]

[0066] The embodiments described above are merely preferred embodiments of the present invention and are not intended to limit the scope of the present invention. Various modifications and improvements made by those skilled in the art to the technical solutions of the present invention without departing from the spirit of the present invention should fall within the protection scope defined by the claims of the present invention.

Claims

1. The use of a compound in the preparation of antifungal and antitumor drugs, characterized in that, The structure of the compound is shown below: ； The fungus is a yeast and / or a filamentous fungus; the tumor is lung cancer and / or breast cancer; The yeast is one or more of Candida glabrata, Cryptococcus neoformans, Candida krusei, Candida albicans, and Candida parapsilosis; the filamentous fungus is one or more of Aspergillus fumigatus, Fusarium, and Mucor.

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