2-[(3-methylphenyl)methylthio]-3,4-dihydropyrimidin-4-one for use in the preparation of a medicament for the prevention and / or treatment of Candida albicans

Abstract

This invention discloses the application of 2-[(3-methylphenyl)methylthio]-3,4-dihydropyrimidin-4-one in the preparation of drugs for the prevention and / or treatment of Candida albicans, belonging to the field of biomedical technology. The compound 2-[(3-methylphenyl)methylthio]-3,4-dihydropyrimidin-4-one is a compound targeting copper homeostasis-related molecules in Candida albicans. At a concentration of 100 μM, it significantly inhibits Candida albicans hyphal formation, and the inhibitory effect is further enhanced at a concentration of 200 μM. Using a large wax moth infection model, it was confirmed that this compound can reduce the virulence of Candida albicans in a dose-dependent manner, and the survival rate of the large wax moth in the 2 mg / kg treatment group was approximately 80%. Simultaneously, this compound can effectively inhibit the colonization of Candida albicans in the host, with a significantly lower bacterial load 48 hours after treatment compared to the control group. This invention provides a new drug candidate for the treatment of Candida albicans infection and has good application prospects.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology, specifically relating to the use of 2-[(3-methylphenyl)methylthio]-3,4-dihydropyrimidin-4-one in the preparation of drugs for the prevention and / or treatment of Candida albicans. Background Technology

[0002] Candida albicans is a symbiotic fungus widely found on the skin and mucous membranes of humans and other mammals. Under specific conditions such as weakened host immunity, prolonged use of antibiotics, immunosuppression, or implanted medical devices, it can transform into an opportunistic pathogen, causing invasive infections. Currently, invasive candidiasis encompasses various serious forms, including candidemia, disseminated infections, and deep infections. It is estimated that Candida albicans causes over 150 million mucosal infections annually in high-risk populations worldwide and results in approximately 200,000 deaths from systemic infections. Currently, there are three classes of first-line treatments for invasive candidiasis, targeting only two pathways—ergosterol synthesis and cell wall activity. Compared to the progress in antimicrobial drug development, the development of new antifungal drugs is severely lagging—only one new class of drug has been launched in the past two decades, while 15 new antibiotics have been approved during the same period. Therefore, a deep understanding of the pathogenic mechanism of Candida albicans is of significant guiding importance for the development of novel drug targets.

[0003] One obstacle that pathogenic microorganisms, including Candida albicans, must overcome within the host is micronutrient stress, a defense strategy known as nutritional immunity. To counter this host defense, fungal pathogens have evolved complex mechanisms to precisely balance the delicate boundaries between essential micronutrients. Copper, as a trace metal element, is one of the most fundamental transition elements. It is a cofactor for important enzymes in Candida albicans, such as cytochrome oxidase and superoxide dismutase, participating in respiration, metal uptake, and antioxidant defense. Current research has found that combining copper with nanotechnology as a nanocomposite material for antibacterial coatings can slow down or even inhibit the growth of fungi and other pathogenic microorganisms. To address this challenge, Candida albicans has also evolved complex and precise copper homeostasis regulation mechanisms. For example, the absence of the copper transporter Ctr1 leads to reduced virulence in Candida albicans, and the absence of the copper efflux pump Crp1 affects its survival in acidic and anaerobic conditions (such as the gastrointestinal tract). These studies suggest that these copper homeostasis-related genes may serve as drug targets, playing a crucial role in the development of novel antifungal drugs. Summary of the Invention

[0004] To address the shortcomings of existing technologies, this invention provides the application of 2-[(3-methylphenyl)methylthio]-3,4-dihydropyrimidin-4-one in the preparation of drugs for the prevention and / or treatment of Candida albicans. This compound can inhibit the growth of Candida albicans hyphae and reduce the yeast-hyphae morphology conversion rate, thereby weakening the toxicity of Candida albicans in the host. It is intended to serve as a candidate drug for the treatment of Candida albicans infection, solving the problems of the increasing number of immunocompromised individuals, the limitations of clinical antifungal drugs, and fungal resistance.

[0005] To address the problems in the existing technology, the technical solution adopted by this invention is as follows: Use of 2-[(3-methylphenyl)methylthio]-3,4-dihydropyrimidin-4-one in the preparation of drugs for the prevention and / or treatment of Candida albicans.

[0006] Preferably, the molecular formula of the 2-[(3-methylphenyl)methylthio]-3,4-dihydropyrimidin-4-one is C2 12 H 12 N2OS, the structure is as follows: .

[0007] Preferably, the drug is a drug that inhibits the growth of Candida albicans hyphae.

[0008] Preferably, the drug is a drug that reduces the virulence of Candida albicans.

[0009] Preferably, the drug is a drug that inhibits the colonization of Candida albicans in the host.

[0010] Preferably, the drug is a compound that targets copper homeostasis-related molecules in Candida albicans. Preferably, the copper stable molecule of Candida albicans is Cox17.

[0011] A pharmaceutical composition for the prevention and / or treatment of Candida albicans infection, said pharmaceutical composition comprising an effective amount of 2-[(3-methylphenyl)methylthio]-3,4-dihydropyrimidin-4-one and a pharmaceutically acceptable carrier.

[0012] Preferably, the dosage of 2-[(3-methylphenyl)methylthio]-3,4-dihydropyrimidin-4-one is 0.5-2 mg / kg.

[0013] Preferably, the dosage of 2-[(3-methylphenyl)methylthio]-3,4-dihydropyrimidin-4-one is 2 mg / kg.

[0014] Beneficial effects: Compared with existing technologies, the application of 2-[(3-methylphenyl)methylthio]-3,4-dihydropyrimidin-4-one in the preparation of drugs for the prevention and / or treatment of Candida albicans is presented in this invention. This compound targets copper homeostasis-related molecules in Candida albicans, unlike traditional antifungal compounds that target ergosterol synthesis or cell wall integrity. This compound interferes with the copper homeostasis of Candida albicans, inhibits hyphal generation in wild-type Candida albicans strains, reduces the virulence of wild-type Candida albicans in a large wax moth infection model, significantly reducing the mortality rate of infected large wax moths, and simultaneously inhibits the colonization ability of Candida albicans in the host. That is, this compound leads to a decrease in the morphological conversion efficiency and / or pathogenicity of Candida albicans, providing a favorable direction for the prevention and treatment of Candida albicans. Attached Figure Description

[0015] Figure 1 This is a diagram showing the inhibitory effect of the compound of the present invention on the hyphal growth of Candida albicans; Figure 2 This is a graph showing the detection results of the compound of the present invention in treating systemic Candida albicans wild-type giant wax moth; Figure 3 The effect of the compound of this invention on the colonization of Candida albicans in the host body; Figure 4 This is a diagram showing the relationship between the compound of this invention and the copper stable molecule Cox17 of Candida albicans. Detailed Implementation

[0016] The present invention will now be described in further detail with reference to specific embodiments. The embodiments given are only for illustrating the present invention and are not intended to limit the scope of the present invention. The experimental methods in the following embodiments are conventional methods commonly known to those skilled in the art and do not constitute a limitation on the present invention in any way.

[0017] Wild-type Candida albicans strain SN152 was donated by Malcolm Whiteway of Concordia University, Canada.

[0018] Example 1: The inhibitory effect of 2-[(3-methylphenyl)methylthio]-3,4-dihydropyrimidin-4-one on Candida albicans mycelial growth To investigate the effect of 2-[(3-methylphenyl)methylthio]-3,4-dihydropyrimidin-4-one on Candida albicans mycelial formation.

[0019] Specifically, the steps were as follows: Wild-type Candida albicans strain SN152 was cultured in YPD liquid medium at 30°C in a shaker at 200 rpm until the stationary phase (approximately 18 hours). The next day, it was transferred at a 1 / 20 ratio to 20% liquid FBS, and the compound was added to achieve final concentrations of 100 μM (containing 0.05% DMSO) and 200 μM (containing 0.1% DMSO), respectively. Controls were established with either 0.05% or 0.1% DMSO solution. The culture was continued at 37°C with shaking for 2 hours. Cells were collected and photographed for observation. The results showed that Candida albicans in the control group could form hyphae normally in 20% FBS, while hyphal formation was significantly limited in the 100 μM compound group. Furthermore, the 200 μM compound further inhibited hyphal formation, almost completely preventing hyphae formation, indicating that the compound has an inhibitory effect on Candida albicans hyphal formation.

[0020] Furthermore, we used Autodock software to detect the interaction between 2-[(3-methylphenyl)methylthio]-3,4-dihydropyrimidin-4-one and the copper stable molecule Cox17 from Candida albicans, and the results are as follows: Figure 4 As shown in the figure, the compound interacts with sites such as Cys16 of Cox17, suggesting that the compound has a targeting effect on copper stability.

[0021] Example 2: The effect of 2-[(3-methylphenyl)methylthio]-3,4-dihydropyrimidine-4-one on the inhibition of Candida albicans virulence in a systemic infection model of *Candida albicans*. To investigate the effect of 2-[(3-methylphenyl)methylthio]-3,4-dihydropyrimidin-4-one on the virulence of Candida albicans in a model of infection with the wax moth.

[0022] Specifically, the steps are as follows: In this embodiment, the wild-type Candida albicans strain SN152 was cultured in YPD liquid medium at 30 °C in a shaker at 200 rpm until the stationary phase (approximately 18 hours). The bacterial cells were collected and counted, and the bacterial concentration was adjusted to 3 × 10⁻⁶. 7 / mL, a certain concentration of DMSO solution of 2-[(3-methylphenyl)methylthio]-3,4-dihydropyrimidin-4-one (containing 3.87% DMSO) was added to make the final compound concentration injected into the large wax moth body 0.5 mg / kg, 1 mg / kg, 2 mg / kg. 10 μL of bacterial solution was drawn up with a microsyringe and injected into the caudal peduncle of the large wax moth larva. The control group was not treated (i.e., large wax moths directly injected with 3.87% DMSO solution). The cells were cultured at 37 ℃ and the survival rate was observed. Those that did not respond to external stimuli were considered dead. The results showed that by day 7, all the wax moths in the control group (without drug injection) had died, with an average survival period of 2 days; the survival rate of the wax moths in the 0.5 mg / kg drug treatment group was approximately 13.3%, with an average survival period of 3 days; the survival rate of the wax moths in the 1 mg / kg drug treatment group was approximately 33.3%, with an average survival period of 6 days; and the survival rate of the wax moths in the 2 mg / kg drug treatment group was approximately 80%. The analysis showed a statistically significant difference, indicating that the compound can reduce the virulence of wild-type Candida albicans in the wax moth infection model.

[0023] Example 3

[0024] Effects of 2-[(3-methylphenyl)methylthio]-3,4-dihydropyrimidin-4-one on the colonization of Candida albicans in the host. To further investigate the antibacterial effect of 2-[(3-methylphenyl)methylthio]-3,4-dihydropyrimidine-4-one, the effect of this compound on the colonization ability of Candida albicans in vivo was examined using a large wax moth infection model.

[0025] Specifically, in this embodiment, the wild-type Candida albicans strain SN152 was cultured in YPD liquid medium at 30°C in a shaker at 200 rpm until the stationary phase (approximately 18 hours). The bacterial cells were collected and counted, and the bacterial concentration was adjusted to 3 × 10⁻⁶. 7 / mL, add 2-[(3-methylphenyl)methylthio]-3,4-dihydropyrimidin-4-one DMSO solution (containing 3.87% DMSO) to make the final compound concentration injected into the large wax moth body 2 mg / kg. 10 μL of bacterial suspension was drawn using a microsyringe and injected into the caudal peduncle of the large wax moth larvae. Large wax moths without drug treatment (i.e., directly injected with 3.87% DMSO solution) served as the control group. Large wax moths from both the control and treatment groups were collected at 24 h and 48 h, respectively. 1 mL of PBS buffer was added and the mixture was homogenized. The homogenate was diluted 100-fold and spread onto YPD+chloramphenicol plates. The results showed that at 24 h, the bacterial load in the control group (no drug injection group) was 6.7 × 10⁻⁶. 5 / animal, while the drug-treated group had only 3.3 × 10 5 / animal; at 48h, the bacterial load of *Gnaphalium affine* in the control group (without drug injection) was 1.1 × 10-1. 6 / animal, while the drug-treated group had only 4.3 × 10 5 / each; analysis showed a statistically significant difference, indicating that the compound can reduce the colonization of Candida albicans in the giant wax moth.

[0026] The present invention has been described above by way of example. It should be noted that any simple modifications, alterations or other equivalent substitutions that can be made by those skilled in the art without creative effort without departing from the core of the present invention fall within the protection scope of the present invention.

Claims

1. 2-[(3-methylphenyl)methylthio]-3,4-dihydropyrimidin-4-one in the preparation of drugs for the prevention and / or treatment of Candida albicans.

2. The application according to claim 1, characterized in that, The molecular formula of the 2-[(3-methylphenyl)methylthio]-3,4-dihydropyrimidin-4-one is C 12 H 12 N2OS, the structure is as follows: .

3. The application according to claim 1, characterized in that, The drug is a drug that inhibits the growth of Candida albicans hyphae.

4. The application according to claim 1, characterized in that, The drug is used to reduce the virulence of Candida albicans.

5. The application according to claim 1, characterized in that, The drug is a compound that targets copper homeostasis-related molecules in Candida albicans.

6. The application according to claim 1, characterized in that, The copper stable molecule of Candida albicans is Cox17.

7. The application according to claim 1, characterized in that, The drug is an inhibitor that inhibits the colonization of Candida albicans in the host.

8. A pharmaceutical composition for the prevention and / or treatment of Candida albicans infection, characterized in that, The pharmaceutical composition comprises an effective amount of 2-[(3-methylphenyl)methylthio]-3,4-dihydropyrimidin-4-one and a pharmaceutically acceptable carrier.

9. The pharmaceutical composition according to claim 7, characterized in that, The dosage of 2-[(3-methylphenyl)methylthio]-3,4-dihydropyrimidin-4-one is 0.5-2 mg / kg.

10. The pharmaceutical composition according to claim 8, characterized in that, The dosage of 2-[(3-methylphenyl)methylthio]-3,4-dihydropyrimidin-4-one is 2 mg / kg.

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