A gold complex with aggregation-induced emission characteristics and a preparation method and application thereof

Abstract

The application discloses a gold complex with an aggregation-induced emission characteristic and a preparation method and application thereof, and provides a compound of formula I, a compound of formula II or a pharmaceutically acceptable salt thereof: The compound of the application has an aggregation-induced emission effect, can effectively avoid self-absorption and interference of biological sample background fluorescence, and provides a fluorescent dye with excellent performance for fungal cell structure staining. The compound of the application can drive fungal iron death through induction of mitochondrial dysfunction and oxidative stress, significantly up-regulate a multi-modal mechanism of activating iron death related to genes of fungal iron transport, iron reduction, iron-sulfur cluster synthesis and assembly and hemoglobin binding, and provide a long-term therapeutic effect for clinical fungal drug-resistant strain prevention and treatment.

Description

Technical Field

[0001] This invention relates to the field of biomedical technology, and in particular to a gold complex with aggregation-induced emission properties, its preparation method, and its application. Background Technology

[0002] Ferroprelation is a regulated form of cell death characterized by iron homeostasis imbalance, lethal accumulation of reactive oxygen species (ROS), depletion of the oxidative stress defense system, and aberrant lipid peroxidation. Growing evidence suggests that ferroprelation plays a crucial regulatory role in numerous diseases, including neurodegenerative diseases (Alzheimer's, Parkinson's, and Huntington's), cancer, stroke, cerebral hemorrhage, ischemia-reperfusion injury, traumatic brain injury, and renal degeneration, making it a promising target for developing novel therapeutic strategies. The molecular mechanisms of ferroprelation in mammalian cells are well-established, and the development of ferroprelation inhibitors or inducers has become a focus in disease treatment, particularly in anti-tumor therapy. Notably, iron-activated ferroprelation-like cell death has been observed in bacteria. However, research on ferroprelation in the antifungal field remains significantly inadequate, with very limited reporting on this topic. In particular, it remains unclear whether ferroprelation can serve as a therapeutic target for fungal infections.

[0003] In recent years, monovalent gold complexes have attracted attention as effective inducers of ferroptosis. Their mechanism of action involves targeting and inhibiting thioredoxin reductase (TrxR) via covalent binding to selenium-sulfur groups (such as Sec498), leading to irreversible enzyme inactivation. TrxR, a key antioxidant enzyme, maintains intracellular redox homeostasis and regulates the antioxidant defense system by reducing thioredoxin (Trx). Inhibition of TrxR impairs ROS clearance, triggering a series of events including antioxidant system collapse, iron metabolism dysregulation, impaired lipid peroxidation repair, and oxidative membrane damage. These findings highlight monovalent gold complexes as a novel strategy for inducing ferroptosis and overcoming tumor resistance. However, unlike tumor cells, fungal cells can develop resistance through gene mutations, metabolic reprogramming, or enhanced antioxidant defenses, potentially undermining the long-term efficacy of single-pathway ferroptosis inducers. Therefore, developing novel drugs that can activate ferroptosis through multimodal mechanisms has significant clinical implications. Summary of the Invention

[0004] The present invention aims to at least solve one of the aforementioned technical problems existing in the prior art. Therefore, the object of the present invention is to provide a gold complex with aggregation-induced emission properties, its preparation method, and its application.

[0005] To achieve the above objectives, the technical solution adopted by the present invention is as follows:

[0006] In a first aspect, the present invention provides a compound of formula I, a compound of formula II, or a pharmaceutically acceptable salt thereof:

[0007]

[0008]

[0009] Wherein, R1, each of R2, and each of R4 are each independently selected from H, halogens, C1-C6 alkyl groups, and C1-C6 alkoxy groups; the alkyl and alkoxy groups are optionally unsubstituted or substituted by one or more R groups. a replace;

[0010] Each R3 is independently selected from halogens, C6–C6. 20 Aryl, C6~C 20 arylphosphinyl; the aryl or arylphosphinyl group may optionally be unsubstituted or substituted with one or more R groups. a replace;

[0011] Each R a Selected from halogens and C1-C6 alkyl groups.

[0012] In some embodiments of the present invention, the halogen is selected from F, Cl, Br, and I.

[0013] In some embodiments of the present invention, the C1-C6 alkyl group is selected from methyl, ethyl, 1-propyl, 2-propyl, 1-butyl, 2-methyl-1-propyl, 2-butyl, 2-methyl-2-propyl, 1-pentyl, 2-pentyl, 3-pentyl, 2-methyl-2-butyl, 3-methyl-2-butyl, 3-methyl-1-butyl, 2-methyl-1-butyl, 1-hexyl, 2-hexyl, 3-hexyl, 2-methyl-2-pentyl, 3-methyl-2-pentyl, 4-methyl-2-pentyl, 3-methyl-3-pentyl, 2-methyl-3-pentyl, 2,3-dimethyl-2-butyl, and 3,3-dimethyl-2-butyl.

[0014] In some embodiments of the present invention, the C1-C6 alkoxy groups are selected from methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, sec-butoxy, isobutoxy, tert-butoxy, n-pentoxy, neopentoxy, isopentoxy, tert-pentoxy, n-hexyloxy, cyclohexyloxy, and neohexyloxy.

[0015] In some embodiments of the present invention, C6 to C 20 The aryl group is selected from benzene ring, naphthyl, and anthracene.

[0016] In some embodiments of the present invention, C6 to C 20 The arylphosphine group is selected from diphenylphosphine and triphenylphosphine.

[0017] In some embodiments of the present invention, each R3 is independently selected from F, Cl, Br, I, benzene ring, biphenyl, diphenylphosphine, and triphenylphosphine; wherein each phenyl group is optionally unsubstituted or substituted with one or more R3 groups. a replace.

[0018] In some embodiments of the present invention, the compound is selected from the following compounds:

[0019]

[0020]

[0021] A second aspect of the present invention provides a method for preparing the aforementioned compound, comprising the following steps:

[0022] Compound I-1 and compound I-2 were subjected to a Suzuki coupling reaction, then reacted with imidazole, followed by a quaternization reaction with compound I-3, and then an exchange reaction to obtain compound I.

[0023]

[0024] Compound II-1 and compound II-2 were subjected to a Suzuki coupling reaction, followed by reaction with imidazole, then quaternization with compound I-3, and finally exchange reaction to obtain compound II.

[0025]

[0026] R1, R2, and R4 are defined as described above; R5 and R6 are each independently selected from any one of borate group, trifluoroborate group, triol borate group, and borate ester group; X, X1, X2, X3, and X4 are each independently selected from halogens.

[0027] In some embodiments of the present invention, there is no particular limitation as to borate group, borate ester group, trifluoroborate group or triol borate group, and for example, the groups shown in the following structural formulas can be exemplified:

[0028]

[0029] In some embodiments of the present invention, the Suzuki coupling reaction is carried out in the presence of a palladium catalyst and a base.

[0030] In some embodiments of the present invention, the palladium catalyst is selected from at least one of tetraphenylphosphine palladium, palladium acetate, and 1,1'-bis(diphenylphosphine)ferrocene palladium dichloride; the molar ratio of the compound of formula I-1 to the palladium catalyst is 1:(0.01-0.2), preferably 1:0.05; the molar ratio of the compound of formula II-2 to the palladium catalyst is 1:(0.01-0.2), preferably 1:0.05.

[0031] In some embodiments of the present invention, the base is selected from one or more of potassium carbonate, sodium carbonate, potassium phosphate, and cesium carbonate. The molar ratio of the compound of formula I-1 to the base is 1:(1-3); the molar ratio of the compound of formula II-2 to the base is 1:(1-3).

[0032] In some embodiments of the present invention, the solvent for the Suzuki coupling reaction is selected from at least one of tetrahydrofuran, N,N-dimethylformamide, dimethyl sulfoxide, toluene, n-hexane, acetonitrile, 1,4-dioxane, and water. The solvent is a mixture of an organic solvent and water, with a volume ratio of organic solvent to water of (3–10):1, preferably 5:1; more preferably, a mixture of tetrahydrofuran and water. The mass-volume ratio of the compound of formula I-1 to the solvent is 1:(30–60), preferably 1:(35–45). The volume ratio of the compound of formula II-2 to the solvent is 1:(30–60), preferably 1:(35–45).

[0033] In some embodiments of the present invention, the Suzuki coupling reaction is carried out under reflux conditions; the Suzuki coupling reaction time is 1 to 8 hours and the temperature is 70°C to 120°C.

[0034] In some embodiments of the present invention, the reaction of the Suzuki coupling reaction product with imidazole is a CN coupling reaction; the CN coupling reaction is carried out in the presence of a copper catalyst and a base; the copper catalyst includes at least one of copper acetate, copper trifluoroacetate, copper trifluoromethanesulfonate, copper acetylacetone, copper sulfate, copper chloride, copper bromide, copper iodide, cuprous oxide, cuprous bromide, cuprous iodide, and cuprous chloride; the molar ratio of the Suzuki coupling reaction product to the copper catalyst is 1:(0.1-0.3), such as 1:0.2; the molar ratio of the Suzuki coupling reaction product to the base is 1:(1-3). The solvent for the CN coupling reaction is selected from at least one of tetrahydrofuran, N,N-dimethylformamide, dimethyl sulfoxide, toluene, n-hexane, acetonitrile, 1,4-dioxane, and water. The mass-to-volume ratio of the Suzuki coupling reaction product to the solvent is 1:(50-80), preferably 1:(55-65); the reaction temperature of the CN coupling reaction is 130-180℃, such as 140-180℃; and the reaction time is 12-48h.

[0035] In some embodiments of the present invention, the reaction temperature of the quaternization reaction is 80–120°C, such as 90–110°C; and the reaction time is 10–24 h.

[0036] In some embodiments of the present invention, the exchange reaction includes anion exchange reaction and ligand exchange reaction.

[0037] In some embodiments of the present invention, the anion exchange reaction is carried out using silver salts, wherein the silver salts include at least one selected from Ag₂O, Ag₂CO₃, AgOAc, and Ag₂S. The anion exchange reaction is carried out under light-protected conditions; the reaction time is 24–72 h.

[0038] In some embodiments of the present invention, the ligand exchange reaction is carried out using a gold complex, which includes at least one of (tht)AuCl, (tht)AuC6F5, (PPh3)AuC6F5, (JohnPhos)AuC6F5, and (IMes)AuC6F5. The reaction time of the ligand exchange reaction is 10–24 h.

[0039] A third aspect of the invention provides a pharmaceutical composition comprising the said compound, and optionally, pharmaceutically acceptable excipients.

[0040] The pharmaceutical compositions of the present invention are suitable for various routes of administration and can therefore be formulated into any pharmaceutically acceptable dosage form. For example, the pharmaceutical compositions described above can be administered to patients or subjects requiring such treatment via oral, parenteral, rectal, or pulmonary administration. For oral administration, the pharmaceutical compositions can be formulated into oral preparations, such as conventional oral solid dosage forms like tablets, capsules, pills, granules, etc.; or into oral liquid preparations, such as oral solutions, oral suspensions, syrups, etc. When formulating oral preparations, suitable fillers, binders, disintegrants, lubricants, etc., can be added. For parenteral administration, the pharmaceutical compositions described above can also be formulated into injections, including injection solutions, sterile powders for injection, and concentrated solutions for injection. When formulating injections, conventional methods in the existing pharmaceutical field can be used; when preparing injections, excipients may not be added, or suitable excipients may be added depending on the properties of the drug. For rectal administration, the pharmaceutical compositions can be formulated into suppositories, etc. For pulmonary administration, the pharmaceutical compositions can be formulated into inhaled preparations, aerosols, powder inhalers, or sprays, etc.

[0041] Pharmaceutically acceptable excipients are substances that are non-toxic, compatible with the active ingredient, and otherwise biologically suitable for use in organisms. The selection of specific excipients will depend on the route of administration or the type and state of disease in the treatment of a particular patient. Examples of pharmaceutically acceptable excipients include, but are not limited to, solvents, diluents, dispersants, suspending agents, surfactants, isotonic agents, thickeners, emulsifiers, binders, lubricants, stabilizers, hydrating agents, emulsification accelerators, buffers, absorbents, colorants, ion exchangers, release agents, coating agents, flavoring agents, and antioxidants, which are common in the pharmaceutical field. Flavoring agents, preservatives, and sweeteners may also be added to the pharmaceutical composition where necessary.

[0042] In some embodiments of the present invention, the pharmaceutical composition further includes at least one antifungal agent.

[0043] In some embodiments of the present invention, the at least one antifungal agent is an azole drug, echinocandin, amphotericin B deoxycholate, amphotericin B cochleate, 5-fluorocytosine, terbinafine, griseofulvin, VL-2397, ibrexafungerp, orotomide F901318, or a combination thereof. In some embodiments of the present invention, the azole drug is ketoconazole, fluconazole, posaconazole, itraconazole, voriconazole, isaconazole, or miconazole. In some embodiments, the echinocandin is caspofungin, anidoxuridine, micafungin, or rezafungin.

[0044] In some embodiments of the present invention, the pharmaceutical composition is a fungal fluorescence imaging agent.

[0045] A fourth aspect of the invention provides the use of the said compound in the preparation of medicaments for the prevention and / or treatment of fungal infections or diseases, and in fluorescent probes.

[0046] In some embodiments of the present invention, the fluorescent probe is used for fungal fluorescence imaging.

[0047] In some embodiments of the present invention, the fungal infection or disease includes infection or disease caused by susceptible fungal strains and / or infection or disease caused by drug-resistant fungal strains.

[0048] In some embodiments of the present invention, the fungal infection to be prevented and / or treated is selected from candidemia, invasive candidiasis, tinea capitis, tinea corporis, tinea pedis, onychomycosis, perionychomycosis, tinea versicolor, oral thrush, fungal keratitis, vaginal candidiasis, respiratory candidiasis, biliary candidiasis, esophageal candidiasis, urinary tract candidiasis, systemic candidiasis, cutaneous and mucosal candidiasis, aspergillosis, mucosomycosis, South American blastomycosis, North American blastomycosis, histoplasmosis, coccidioidomycosis, sporotrichosis, fungal sinusitis, or chronic sinusitis. In some embodiments, the infection is candidemia or invasive candidiasis.

[0049] In some embodiments of the present invention, the fungal infection to be prevented and / or treated includes infection with at least one of the following: Candida albicans, Candida glabrata, Candida dubliniensis, Candida krusei, Candida parapsilosis, Candida tropicalis, Candida orthopsilosis, Candida guilliermondii, Candida rugosa, Candida auris, Candida lusitaniae, Aspergillus fumigatus, Aspergillus flavus, Aspergillus terreus, Aspergillus niger, Aspergillus candidus, Aspergillus clavatus, or Aspergillus ochraceus.

[0050] In some embodiments of the present invention, the fungal infection to be prevented and / or treated includes at least one infection of Candida albicans, Candida krusei, Candida glabrata, Candida tropicalis, and Candida parapsilosis.

[0051] In some embodiments of the present invention, the drug induces fungal iron death and drives fungal mitochondrial dysfunction (such as tricarboxylic acid cycle and / or oxidative phosphorylation disorders, decreased membrane potential and / or ATP levels, oxidative stress).

[0052] In some embodiments of the invention, the drug induces fungal iron death by inhibiting TrxR activity.

[0053] In some embodiments of the present invention, the drug is used in photodynamic therapy, wherein the light conditions for photodynamic therapy are: light intensity 0.01–2.0 W / cm².2 The illumination time is 10 to 60 minutes; preferably, the light source is white light of 300 to 700 nm.

[0054] The beneficial effects of this invention are:

[0055] The compounds of this invention possess aggregation-induced emission effects, which can effectively avoid interference from self-absorption and background fluorescence of biological samples, providing a high-performance fluorescent dye for staining fungal cell structures.

[0056] The compounds of this invention have good biocompatibility and excellent antibacterial effects against a variety of fungi. Under light conditions, the antibacterial effect is significantly enhanced, playing a synergistic role in chemophotodynamic antifungal activity.

[0057] The compounds of this invention can activate ferroptosis in fungi through multimodal mechanisms, such as inducing mitochondrial dysfunction and oxidative stress to drive ferroptosis, and significantly upregulating genes related to fungal iron transport, iron reduction, iron-sulfur cluster synthesis and assembly, and heme binding, providing a long-term effective solution for the prevention and control of drug-resistant fungal strains in clinical practice. Attached Figure Description

[0058] Figure 1 The images show the UV-Vis absorption spectra of complexes G1 to G5 in Experimental Example 1 of this invention; where a and b are the UV-Vis absorption spectra of G1 and G2 in THF / Hex mixed solutions at 0% and 90% Hex content, respectively; c and d are the UV-Vis absorption spectra of G3 and G4 in THF / Hex mixed solutions at 0% and 99% Hex content, respectively; and e is the UV-Vis absorption spectrum of G5 in THF / Hex mixed solutions at 0% and 70% Hex content, respectively.

[0059] Figure 2 The complexes G1 to G5 in Experimental Example 1 of this invention (corresponding to respectively) Figure 2 Fluorescence spectra of (a, b, c, d, e) under THF / Hex mixed solution conditions.

[0060] Figure 3 In this paper, a represents the ROS generation capacity of complexes G1 to G5 in Test Example 1 of the present invention; b represents the ability of complexes G1 to G5 in Test Example 1 of the present invention to generate singlet oxygen.

[0061] Figure 4 The in vitro imaging effect of complexes G1 to G5 of the present invention on fungi (standard strains) in Example 6 of the present invention.

[0062] Figure 5 The co-localization effect of complex G3 with commercial dye in Example 7 of the present invention (a), as well as the dye imaging photostability (b) and imaging no-wash staining (c).

[0063] Figure 6 The in vitro antifungal (standard strain) effects of complexes G1 to G5 in Example 8 of this invention are shown in Figure a, which is the antibacterial result on plate, and Figures b and c are the results of scanning electron microscopy and transmission electron microscopy, respectively.

[0064] Figure 7 In vitro imaging (a) and anti-drug-resistant fungal strain effects (b) of complexes G1-G5 in Example 10 of the present invention.

[0065] Figure 8 This study investigates the toxicity of complexes G1 to G5 in HaCaT cells (a) and L929 cells (b) in Experimental Example 2 of this invention.

[0066] Figure 9 This is a lipidomics analysis of the mechanism of fungal death induced by complex G3 in Example 11 of the present invention; a and b are OPLS-DA analysis of differentially expressed genes before and after G3 treatment; c to e are lipid types and lipid unsaturation analysis; f is differential lipid analysis.

[0067] Figure 10 This is to verify the effect of complex G3 in TrxR activity inhibition in inducing fungal ferroptosis in Example 12 of the present invention; a shows the changes in antibacterial effect after treatment with different cell death inhibitors; b and c show the qualitative and quantitative analysis of lipid peroxidation levels in Candida albicans under different treatment conditions; d shows the analysis of Gpx activity levels in the bacteria; e shows the qualitative analysis of reactive oxygen species levels in Candida albicans.

[0068] Figure 11 This is to verify the effect of complex G3 in inducing fungal iron death by inhibiting TrxR activity in Example 12 of the present invention; a is the analysis of TrxR activity level in the fungus; b is the quantitative analysis of reactive oxygen species level in Candida albicans; c to e are the assessment of cell viability and lipid peroxidation level after TRi-1 inhibits TrxR, respectively.

[0069] Figure 12 This invention validates the mitochondrial oxidative stress-induced ferroptosis induced by complex G3 in Examples 13-15; a and b are GSEA analyses from transcriptome sequencing, representing downregulation of the TCA cycle and oxidative phosphorylation pathway, respectively; c and d are mitochondrial membrane potential and ATP levels before and after G3 treatment; e is the detection of mitochondrial ROS levels; f and g are the effects of treatment with the mitochondrial reactive oxygen species scavenger Mito-TEMPO on G3-induced lipid peroxidation and antifungal effects, respectively.

[0070] Figure 13To verify the effect of complex G3 on the fungal mitochondrial-iron cascade reaction in Example 16 of this invention, a is GO enrichment analysis; b is iron staining in Candida albicans; c-f are the gene expression of fungal iron transport, iron reduction, iron-sulfur cluster synthesis and assembly, and heme binding, respectively.

[0071] Figure 14 For the validation of the mouse skin wound fungal infection model in Example 17 of this invention, a and b represent the recovery status of mouse skin wounds; c and h represent HE staining and quantification of granulation tissue; d and i represent Masson staining and collagen quantification results; e, f, j, and k represent the staining and quantification results of IL-6 and TNF-α, respectively; g and l represent the staining and quantification of the neovascularization marker CD31, respectively.

[0072] Figure 15 For the biosafety evaluation of mice in Examples 17 and 18 of this invention, a is HE staining of mouse heart, liver, spleen, lung and kidney tissues; b to e are mouse blood biochemical results; f is mouse weight record; g and h represent the hemolysis experiment of complex G3.

[0073] Figure 16 The treatment effect of the mouse fungal keratitis infection model in Example 19 of this invention is shown in Figure 19. a and d represent the corneal recovery status and clinical grade score of the mice; b and e represent the sodium fluorescein staining and quantification of the mouse cornea; c and f represent the fungal load on the mouse cornea; g represents the HE staining of the cornea before and after treatment.

[0074] Figure 17 The treatment effect of the mouse systemic infection model in Example 20 of the present invention is shown in Figure 1; a is the survival rate curve of mice; b to f represent the bacterial load in mouse organs; g is HE staining of major organs.

[0075] Figure labels: * indicates p<0.05; ** indicates p<0.005; *** indicates p<0.001. Detailed Implementation

[0076] The present invention will be further described in detail below through specific embodiments. Unless otherwise specified, the raw materials, reagents, or apparatus used in the embodiments and comparative examples are all available from conventional commercial sources or can be obtained by existing technical methods. Unless otherwise specified, the test or experimental methods are conventional methods in the art.

[0077] The term "pharmaceutical acceptable" means that a drug is chemically or physically compatible with other components that make up a drug dosage form and is physiologically compatible with receptors.

[0078] As used herein, the term "salt" refers to any pharmaceutically acceptable salt commonly used in the pharmaceutical industry, such as non-toxic acid addition salts, metal salts, or metal complexes. Examples of acid addition salts include organic acids (e.g., acetic acid, lactic acid, palmitic acid, maleic acid, citric acid, cholic acid, capric acid, caprylic acid, lauric acid, glutaric acid, glucuronic acid, glyceric acid, glycocholic acid, glyoxylic acid, isocitric acid, isovaleric acid, lactic acid, malic acid, oxaloacetic acid, oxalosuccinic acid, propionic acid, pyruvic acid, ascorbic acid, succinic acid, benzoic acid, palmitic acid, succinic acid, salicylic acid, tartaric acid, methanesulfonic acid, toluenesulfonic acid, and trifluoroacetic acid) and inorganic acids (e.g., hydrochloric acid, hydrobromic acid, sulfuric acid, and phosphoric acid). Representative alkali metal or alkaline earth metal salts include sodium, lithium, potassium, calcium, and magnesium salts.

[0079] Example 1

[0080] In this embodiment, gold complex G1 was prepared. The specific process is as follows:

[0081]

[0082] 1. Synthesis of compound 1c

[0083] Compounds 1a (1.47 g, 5.00 mmol), 1b (1.45 g, 5.00 mmol), Pd(PPh3)4 (0.28 g, 0.25 mmol), and K2CO3 (1.38 g, 10.00 mmol) were added to a 100 mL reaction flask. A solvent system consisting of THF (50 mL) and H2O (10 mL) was then added to the flask. The reaction mixture was refluxed in an oil bath at 80 °C, and the reaction progress was monitored by thin-layer chromatography (TLC) until completion. After cooling to room temperature, the crude product was extracted and separated, and then purified by silica gel column chromatography to give the target compound 1c as an orange solid (1.05 g, 46% yield).

[0084] 2. Synthesis of compound 1e

[0085] 1c (0.82 g, 2.00 mmol), 1d (0.16 g, 2.40 mmol), CuSO4·5H2O (0.10 g, 0.40 mmol), and K2CO3 (0.36 g, 2.60 mmol) were added to a 100 mL reaction flask and combined. Then, 50 mL of DMF was added to the flask, and the mixture was heated under reflux for 24 hours. After the reaction was complete, the mixture was cooled, and a large amount of ice water was added, precipitating a large amount of orange solid 1e. The solid was collected by filtration and dried in a vacuum oven at 50 °C for 24 hours. The crude product could be used directly in the next reaction without further purification.

[0086] 3. Synthesis of compound 1f

[0087] 1e (2.23 g, 5.00 mmol) was added to a 100 mL reaction flask. Iodomethane (1.42 g, 10.00 mmol) and DMF (50 mL) were then added to the flask. The mixture was refluxed at 100 °C for 12 hours. After the reaction was complete, the mixture was cooled and a large amount of ice water was added, precipitating a large amount of orange solid. The solid was collected by filtration and purified by column chromatography to give red solid 1f (2.05 g, 70% yield). 1 H NMR (400MHz, CDCl3) δ10.60 (s, 1H), 8.51 (d, J=6.4Hz, 1H), 8.37 (s, 1H), 7.92 (s, 1H), 7.82 (t, J=6.0H z, 3H), 7.30 (t, J=7.6Hz, 4H), 7.17 (d, J=8.4Hz, 5H), 7.13 (s, 1H), 7.09 (t, J=7.6Hz, 2H), 4.37 (s, 3H). 13 C NMR (101MHz, CDCl3) δ154.0, 149.1, 148.0, 147.0, 136.6, 136.0, 130.2, 12 9.5, 128.5, 126.3, 125.3, 124.3, 124.2, 123.9, 123.3, 122.4, 122.0, 38.1.

[0088] 4. Synthesis of gold complex G1:

[0089] Add 1f (0.18 g, 0.30 mmol) and Ag₂O (0.04 g, 0.15 mmol) to a 50 mL reaction flask. Then introduce DCM (30 mL) into the flask. Stir the mixture at room temperature in the dark for 48 hours. Then add (tht) AuCl (0.10 g, 0.30 mmol) and continue stirring for 12 hours. Filter the reaction mixture through diatomaceous earth, and remove the solvent from the filtrate under reduced pressure. Purify the crude product by rapid column chromatography on neutral alumina using DCM as the eluent, and then recrystallize from the DCM / hexane mixture to give the desired red solid G1 (0.15 g, 70% yield). 1 H NMR (400MHz, CDCl3) δ8.23 (d, J=7.6Hz, 1H), 7.85 (d, J=8.4Hz, 2H), 7.78 (d, J=7.6Hz, 1H), 7. 62 (s, 1H), 7.31 (t, J=7.6Hz, 4H), 7.20 (d, J=8.0Hz, 7H), 7.09 (t, J=7.2Hz, 2H), 4.02 (s, 3H). 13C10 NMR (101 MHz, CDCl3) δ 172.2, 154.1, 148.8, 147.2, 135.4, 130.1, 129.5, 129.4, 128.3, 126.7, 126.3, 125.2, 123.7, 123.1, 122.3, 122.0, 38.9. High-resolution mass spectrometry: m / z: [M+Na] + Calculated value C 28 H 21 AuClN5NaS + : 714.095; Measured value: 714.0757.

[0090] Example 2

[0091] In this embodiment, gold complex G2 was prepared. The specific process is as follows:

[0092]

[0093] 1. Synthesis of compound 2a:

[0094] Benzothiadiazole (3.00 g, 22.00 mmol) was dissolved in 48% hydrobromic acid (20 mL). Bromine (0.84 mL, 16.30 mmol) was added dropwise over 1 hour using a constant-pressure dropping funnel. The mixture was heated to 150 °C and refluxed for 2 hours. After cooling, the reaction mixture was poured into water (500 mL), the solid was filtered, and purified by column chromatography to give the desired white solid 2a (3.30 g, 70% yield). 1 H NMR (600MHz, CDCl3) δ7.97 (d, J=8.5Hz, 1H), 7.85 (d, J=6.8Hz, 1H), 7.48 (t, J=7.4Hz, 1H).

[0095] 2. Synthesis of compound 2b:

[0096] Add 2a (1.08 g, 5.00 mmol), pinacol diborane (1.52 g, 6.00 mmol), Pd(dppf)Cl2 (0.18 g, 0.25 mmol), and KOAc (1.47 g, 15.00 mmol) to a 250 mL reaction flask. Then add 1,4-dioxane (100 mL). Reflux the mixture at 105 °C for 24 hours. After the reaction is complete, remove the solvent by evaporation under reduced pressure. Purify the crude product by column chromatography to give the desired yellow solid 2b (0.67 g, 51% yield). 1H NMR (600MHz, CDCl3) δ8.18 (d, J=6.2Hz, 1H), 8.12 (d, J=8.5Hz, 1H), 7.63-7.56 (m, 1H), 1.45 (s, 12H).

[0097] 3. Synthesis of compound 2d:

[0098] Compounds 2b (0.26 g, 1.00 mmol), 2c (0.81 g, 2.00 mmol), Pd(PPh3)4 (0.06 g, 0.05 mmol), and K2CO3 (0.28 g, 2.00 mmol) were added to a 50 mL reaction flask. A mixture of tetrahydrofuran (THF) (20 mL) and water (2 mL) was added to the flask, and the mixture was refluxed at 80 °C for 24 hours, with the reaction progress monitored by thin-layer chromatography (TLC). After cooling, the mixture was extracted and separated. The target product was purified by silica gel column chromatography to give a red solid 2d (0.26 g, 56% yield). ¹H NMR (600 MHz, CDCl₃) δ 7.96 (s, ¹H), 7.85 (d, J = 8.0 Hz, 2H), 7.66 (s, 2H), 7.37 (d, J = 8.2 Hz, 2H), 7.30 (t, J = 7.0 Hz, 2H), 7.20 (d, J = 7.6 Hz, 2H), 7.17 (d, J = 8.0 Hz, 2H), 7.09 (t, J = 7.4 Hz, 1H), 7.05 (d, J = 7.8 Hz, 2H). Carbon nuclear magnetic resonance (¹³C NMR) (101 MHz, CDCl₃) δ 155.75, 153.47, 147.64, 147.07, 146.69, 133.93, 132.38, 131.51, 130.18, 129.78, 129.59, 127.10, 125.90, 125.11, 123.87, 123.30, 120.11, 115.56.

[0099] 4. Synthesis of compound 2e:

[0100] 2d (0.82 g, 2.00 mmol), 1d (0.16 g, 2.40 mmol), CuSO4·5H2O (0.10 g, 0.40 mmol), and K2CO3 (0.36 g, 2.60 mmol) were added to a 100 mL reaction flask. Then, 50 mL of DMF was added, and the mixture was heated under reflux for 24 hours. After the reaction was complete, the mixture was cooled, and a large amount of ice water was added to precipitate a large amount of orange solid 2e. The solid was collected by filtration and dried in a vacuum oven at 50 °C for 24 hours. The crude product could be used directly in the next reaction without further purification.

[0101] 5. Synthesis of compound 2f

[0102] 2e (2.23 g, 5.00 mmol) was added to a 100 mL reaction flask, followed by methyl iodoform (1.42 g, 10.00 mmol) and DMF (50 mL). The mixture was refluxed at 100 °C for 12 hours. After the reaction was complete, the mixture was cooled and a large amount of ice water was added to precipitate a large amount of orange solid. The solid was collected by filtration and purified by column chromatography to give a red solid 2f (2.32 g, 79% yield). 1 H NMR (600MHz, CDCl3) δ9.00 (s, 1H), 7.91 (dd, J = 24.6, 7.2Hz, 3H), 7.66 (d, J = 7.6Hz, 2H), 7.59 (s, 1H), 7.57 (s,1H),7.47(d,J=8.2Hz,2H),7.34(t,J=6.8Hz,2H),7.30-7.18(m,6H),7.18-7.12(m,1H),4.19(s,3H). 13 C NMR (101MHz, CDCl3) δ155.7,153.4,149.7,146.7,146.3,135.6,133.6,133.0,130.5,13 0.0,129.8,127.5,127.5,126.1,125.1,124.7,124.4,123.4,123.0,121.0,120.4,37.8.

[0103] 6. Synthesis of gold complex G2:

[0104] 2f (0.18 g, 0.30 mmol) and Ag₂O (0.04 g, 0.15 mmol) were added to a 50 mL reaction flask, followed by the addition of DCM (30 mL). The mixture was stirred at room temperature in the dark for 48 hours. Then, (tht) AuCl (0.10 g, 0.30 mmol) was added, and stirring was continued for 12 hours. The reaction mixture was filtered through diatomaceous earth, and the solvent was removed from the filtrate under reduced pressure. The crude product was purified by rapid column chromatography with neutral alumina using DCM as the eluent, followed by recrystallization from a DCM / n-hexane mixture to give the desired red solid G₂ (0.14 g, 67% yield). 1H NMR (400MHz, CDCl3) δ8.01-7.98(m,1H),7.92(d,J=8.4Hz,2H),7.71-7.68(m,2H),7.61(d,J=8.8H z, 2H), 7.37 (t, J = 7.6Hz, 2H), 7.32-7.25 (m, 8H), 7.16 (t, J = 7.4Hz, 1H), 7.11 (s, 1H), 4.05 (s, 3H). 13 C NMR (101MHz, CDCl3) δ170.8,162.1,155.7,153.4,148.4,147.2,146.7,133.8,132.7,13 2.3,130.3,129.8,127.3,125.9,125.6,124.5,124.3,123.1,122.0,121.9,120.2,38.7. HRMS:m / z:[M+Na] + calcd for C 28 H 21 AuClN5NaS + :714.095; found:714.0762.

[0105] Example 3

[0106] In this embodiment, gold complex G3 was prepared. The specific process is as follows:

[0107]

[0108] 1. Synthesis of compound 3b:

[0109] Compounds 3a (1.75 g, 5.00 mmol), 1b (1.46 g, 5.00 mmol), Pd(PPh3)4 (0.28 g, 0.25 mmol), and K2CO3 (1.38 g, 10.00 mmol) were added to a 100 mL reaction flask. A solvent system of THF (50 mL) and H2O (10 mL) was then added. The reaction mixture was refluxed in an oil bath at 80 °C, and the reaction progress was monitored by TLC until completion. After cooling to room temperature, the crude product was extracted and purified by silica gel column chromatography to give the target compound 3b as an orange solid (1.03 g, 40% yield).

[0110] 2. Synthesis of compound 3c:

[0111] 3b (1.03 g, 2.00 mmol), 1d (0.16 g, 2.40 mmol), CuSO4·5H2O (0.10 g, 0.40 mmol), and K2CO3 (0.36 g, 2.60 mmol) were added to a 100 mL reaction flask. DMF (50 mL) was then added, and the mixture was heated under reflux for 24 hours. After the reaction was complete, the mixture was cooled, and a large amount of ice water was added to precipitate a large amount of orange solid 3c. The solid was collected by filtration and dried in a vacuum oven at 50 °C for 24 hours. The crude product was used directly in the next reaction without further purification.

[0112] 3. Synthesis of compound 3d:

[0113] 3c (2.53 g, 5.00 mmol) was added to a 100 mL reaction flask, followed by methyl iodoform (1.42 g, 10.00 mmol) and DMF (50 mL). The mixture was refluxed at 100 °C for 12 hours. After the reaction was complete, the mixture was cooled and a large amount of ice water was added to precipitate a large amount of orange solid 3d. The solid was collected by filtration and purified by column chromatography to give red solid 3d (2.10 g, 65% yield). 1 H NMR (600MHz, CDCl3) δ 10.82 (s, 1H), 8.52 (s, 1H), 8.34 (s, 1H), 7.73 (d, J = 70.6Hz, 4H), 7.06 (d, J = 89.8Hz, 10H), 4.34 (s, 3H), 3.81 (s, 6H). 13 C NMR (151MHz, CDCl3) δ156.56,154.11,150.00,148.05,139.97,137.00,136.50,130.11, 127.43,126.58,125.84,124.45,123.60,122.87,122.45,118.96,114.91,55.53,37.77.

[0114] 4. Synthesis of complex G3:

[0115] 3d (0.19 g, 0.30 mmol) and Ag₂O (0.04 g, 0.15 mmol) were added to a 50 mL reaction flask, followed by the addition of DCM (30 mL). The mixture was stirred at room temperature in the dark for 48 hours. Then, AuCl (0.10 g, 0.30 mmol) was added, and stirring was continued for 12 hours. The reaction mixture was filtered through diatomaceous earth, and the solvent was removed from the filtrate under reduced pressure. The crude product was purified by rapid column chromatography with neutral alumina using DCM as the eluent, followed by recrystallization from a DCM / n-hexane mixture to give the desired red solid G₃ (0.12 g, 53% yield). 1 H NMR (600MHz, CDCl3) δ8.20(d,J=7.6Hz,1H),7.77(dd,J=32.2,8.2Hz,3H),7.61(d,J=1.9Hz,1H),7.21(d,J =1.9Hz,1H),7.16(d,J=8.9Hz,4H),7.04(d,J=8.7Hz,2H),6.88(d,J=8.9Hz,4H),4.01(s,3H),3.82(s,6H). 13 C NMR (151MHz, CDCl3) δ172.22,156.43,154.12,150.55,149.67,140.21,135.66,130.00,127.90,127.35,12 6.75,125.79,123.10,121.92,119.23,114.87,70.58,55.53,38.86,31.60,22.66,14.13.HRMS:m / z:[M+Na] + calcd for C 30 H 25 AuClN5O2NaS + :774.1161; found:774.0986.

[0116] Example 4

[0117] In this embodiment, gold complex G4 was prepared. The specific process is as follows:

[0118]

[0119] 1f (0.18 g, 0.30 mmol) and Ag₂O (0.04 g, 0.15 mmol) were added to a 50 mL reaction flask, followed by the addition of DCM (30 mL). The mixture was stirred at room temperature in the dark for 48 hours. Then, NaCl (0.18 g, 0.30 mmol) and (tht) AuC₆F₅ (0.14 g, 0.30 mmol) were added, and the mixture was stirred overnight at room temperature. The solution was filtered through diatomaceous earth, and the solvent was removed from the filtrate under reduced pressure. The crude product was purified by rapid column chromatography with neutral alumina using DCM as the eluent, followed by recrystallization from a DCM / n-hexane mixture to give the desired red solid G₄ (0.19 g, 75% yield). 1 H NMR (400MHz, CDCl3) δ8.50(d,J=7.6Hz,1H),7.90-7.78(m,4H),7.31(t,J=7.6Hz,4H),7.21(dd,J=8.6,3.8Hz,7H),7.09(t,J=7.2Hz,2H),4.11(s,3H). 13 C NMR (101MHz, CDCl3) δ173.1,154.1,150.3,148.7,147.3,135.0,130.2,12 9.6,129.5,128.4,126.3,126.2,125.2,123.6,123.0,122.5,121.9,38.6. 19 F NMR(376MHz, CDCl3)δ-116.38,-159.73,-163.06.HRMS:m / z:[M+Na] + calcd for C 34 H 21 AuF5N5NaS + :846.1182; found:846.0911.

[0120] Example 5

[0121] In this embodiment, gold complex G5 was prepared. The specific process is as follows:

[0122]

[0123] G2 (0.14 g, 0.20 mmol), AgPF6 (0.05 g, 0.20 mmol), and 5a (0.05 g, 0.20 mmol) were added to a 50 mL reaction flask, followed by the addition of DCM (30 mL). The mixture was stirred at room temperature for 24 hours. The reaction mixture was filtered through diatomaceous earth, and the solvent was removed from the filtrate under reduced pressure. The crude product was purified by recrystallization from a DCM / n-hexane mixture to give the desired red solid G5 (0.17 g, 76% yield). 1 H NMR (400MHz, CD2Cl2) δ7.92(d,J=3.2Hz,1H),7.91(d,J=3.2Hz,1H),7.81(d,J=8.6Hz,2H),7.62(q,J=3.8Hz,2H),7.52-7.47(m,3H),7.44 -7.39(m,9H),7.36(s,2H),7.35(s,1H),7.32(d,J=2.0Hz,1H),7.27(d,J=8.0Hz,2H),7.25-7.22(m,2H),7.13-7.08(m,7H),3.95(s,3H). 13 C NMR(101MHz,CD2Cl2)δ155.8,148.8,147.0,146.6,134.2,134.0,132.4,130.4,130.4,129.8,129.7,12 9.7,129.6,128.4,127.8,127.2,127.1,125.8,125.7,125.6,124.2,124.1,123.1,122.9,120.3,38.5. 31 P NMR(160MHz,CD2Cl2)δ40.24(s,PPh3),-144.52(m,PF6 - ).HRMS:m / z:[M-PF6) + calcd forC 46 H 36 AuN5PS + :918.2173; found:918.2085.

[0124] Experimental Example 1

[0125] 1. Using DMF as solvent, prepare a solution with a final concentration of 2×10⁻⁶. -5 mol·L -1 Solutions of complexes G1 to G5 were placed in cuvettes, and their UV-absorption spectra were detected using a Varian CARY 50 UV spectrometer. Figure 1In the figure, a and b are the UV-Vis absorption spectra of G1 and G2 in THF / Hex mixed solution at 0% and 90% Hex content, respectively. Figure 1 c and d are the UV-Vis absorption spectra of G3 and G4 in THF / Hex mixed solution at 0% and 99% Hex content, respectively; Figure 1 In the figure, e represents the UV-Vis absorption spectra of G5 in a THF / Hex mixed solution at 0% and 70% Hex content, respectively. It can be seen that complexes G1–G5 exhibit distinct absorption peaks in the 400–500 nm range.

[0126] 2. Based on their absorption spectra, fluorescent complexes G1 to G5 were excited by light at 445 nm, 410 nm, 488 nm, 465 nm, and 400 nm, respectively. Figure 2 For coordination compounds G1 to G5 (corresponding to respectively) Figure 2 The fluorescence spectra of (a, b, c, d, e) under THF / Hex mixed solution conditions show that the luminescence of the resulting system gradually increases with the addition of the undesirable solvent Hex, and reaches its maximum value when the Hex content is 90 vol%, 90 vol%, 99 vol%, 99 vol%, and 70 vol%, respectively.

[0127] 3. DCFH-DA (2',7'-Dichlorodihydrofluorescein diacetate) was used as a ROS (reactive oxygen species) detection probe to detect the ROS generation in solution (aggregated state) of the synthesized fluorescent G1–G5 samples. DCFH-DA (0.05 mL, 10 mM) was dissolved in 2 mL of 10 mM sodium hydroxide solution and hydrolyzed with continuous stirring (25 °C, 30 min). The solution was then neutralized with 10 mL of 0.01 M phosphate-buffered saline (PBS, pH 7.4) to obtain a 40 μM DCFH solution. This solution was mixed with 10 μL of the test samples (G1–G5, rose red RB, or Ce6, all at 10 μM) in a quartz cuvette. White light (0.06 W / cm²) was used for analysis. 2 The sample was irradiated, and the fluorescence intensity was measured alternately using a fluorescence spectrophotometer (Horiba Fluoromax4) at an excitation wavelength of 488 nm and an emission wavelength of 525 nm. The results are as follows: Figure 3 As shown in Figure a, under white light irradiation, all complexes except G2 and G5 can generate ROS efficiently and rapidly, significantly outperforming the commercial photosensitizer rose red (RB). Complex G3 exhibits the highest ROS generation efficiency, with fluorescence intensity far exceeding that of other complexes and commercial dyes within the first 60 seconds, indicating its highest generation efficiency, followed by complex G1.

[0128] 4. Singlet Oxygen Detection (Anthracene Dimethylmalonate (ABDA) Method): ABDA (9,10-anthracene-diyl-bis(methylene)dimalonic acid) is used as the singlet oxygen detection probe. Prepare a 10 mM stock solution by dissolving ABDA in PBS. Take 10 μL and mix it with complexes G1–G5, RB, or Ce6 (all at a final concentration of 10 μM) in PBS (final volume 2 mL). Transfer the mixture to a quartz cuvette. Irradiate with white light (0.06 W / cm²). 2 Afterwards, the absorbance change of ABDA at 378 nm was monitored using a UV-Vis spectrophotometer (Varian CARY 50), and its decay rate reflected the singlet oxygen generation efficiency. The results are as follows... Figure 3 As shown in Figure b, under white light irradiation, complexes G3 and G5 can completely oxidize ABDA within the first 60 seconds, reducing its absorption to the baseline standard, while commercial dyes RB and Ce6 require about 120 seconds to completely oxidize ABDA. This indicates that G3 and G5 have a significantly stronger ability to generate singlet oxygen than RB and Ce6.

[0129] Example 6

[0130] This embodiment utilizes complexes G1-G5 to perform in vitro imaging of fungi (standard strains). The specific process is as follows:

[0131] Five clinically relevant Candida species were selected (Candida albicans, Candida krusei, Candida glabrata, Candida tropicalis, and Candida parapsilosis). All strains were provided by Nanfang Hospital, Southern Medical University. The bacterial cells were cultured in YPD medium (37℃, 200 rpm) with shaking for 16 hours, centrifuged (4000×g, 5 min), and resuspended in a solution containing G1–G5 (10 μM, 500 μL PBS) and incubated at 37℃ for 30 min. 1 μL of the stained bacterial suspension was added to a glass slide, covered with a coverslip, and observed using a confocal microscope (Carl Zeiss, LSM 880 with airscan) (excitation wavelength 488 nm, emission wavelength 550–650 nm). Results are as follows: Figure 4 As shown, all complexes exhibit significant fluorescence imaging performance against common clinical fungi, including Candida albicans, Candida krusei, Candida glabrata, Candida tropicalis, and Candida parapsilosis. This invention can provide a novel fluorescence detection method for clinical fungal testing.

[0132] Example 7

[0133] This embodiment utilizes the co-localization effect of complex G3 with commercial dyes and the dye imaging performance to study the process, which is as follows:

[0134] Following the staining with complex G3, the bacterial cells were further incubated with the commercial mitochondrial dye MitoTracker Deep Red (100 nM) at 37°C for 20 minutes. The fluorescence signals of complex G3 (excitation / emission wavelength: 488 / 550–630 nm) and MitoTracker (excitation / emission wavelength: 580 / 630–650 nm) were detected using a laser confocal microscope, respectively, to visualize the fungal structure and the localization of mitochondria. Figure 5 As shown in Figure a, the complex G3 is mainly localized to the cellular lipids and mitochondria of Candida albicans, and exhibits strong co-localization with the dyes BODIPY (Pearson's coefficient 0.81) and Mito-tracker (Pearson's coefficient 0.73).

[0135] Light stability, no-wash performance: such as Figure 5 As shown in Figures b and c, after 300 seconds of continuous laser bleaching, the fluorescence intensity of G3 remained stable, while the fluorescence intensity of the commercial dye BODIPY493 / 503 decreased by more than 15%, indicating that G3 has excellent photostability. After staining Candida albicans with G3 or BODIPY493 / 503 for 30 minutes, centrifugation, and direct laser confocal imaging, G3-imprinted Candida albicans showed high image contrast with negligible background fluorescence, while BODIPY493 / 503 showed some background fluorescence interference. This invention provides a high-performance fluorescent dye for staining fungal cell structures.

[0136] Example 8

[0137] This embodiment utilizes complexes G1-G5 for in vitro antifungal activity (standard strains), and the specific process is as follows:

[0138] Five clinically relevant Candida species were selected (Candida albicans, Candida krusei, Candida glabrata, Candida tropicalis, and Candida parapsilosis). All strains were provided by Nanfang Hospital, Southern Medical University. Single colonies were incubated in YPD medium at 37°C for 16 hours until the logarithmic growth phase. The bacterial suspension concentration was adjusted to 1×10⁻⁶. 6 CFU / mL, incubated with G1–G5 (0–10 μM, 500 μL PBS) at 37°C for 30 minutes. After treatment, each sample was irradiated with white light (0.06 W / cm²). 2 (30 minutes) or protected from light (control). Spread 20 μL of bacterial suspension onto YPD agar plates and incubate at 37°C for 24 hours. Count the colony-forming units (CFU). The bactericidal efficiency is calculated using the formula: Bactericidal efficiency (%) = (AB) / A × 100% (A: average CFU of the control group; B: average CFU of the treatment group). Results are as follows... Figure 6As shown in Figure a, for the standard strains, G1–G5 all exhibited certain antifungal activity (achieving almost 100% antifungal efficiency at concentrations below 10 μM), with G3 showing the strongest antifungal activity (achieving 100% antifungal efficiency at a concentration of 7.5 μM for *Candida glabrata*, *Candida parapsilosis*, *Candida albicans*, and *Candida tropicalis*). Furthermore, compared to dark conditions, the antifungal effect of the complexes was significantly enhanced after white light irradiation (achieving 100% antifungal efficiency at a concentration of 2.5 μM for G3), indicating a strong photodynamic effect.

[0139] Example 9

[0140] This embodiment utilizes complex G3 to disrupt the structure of Candida albicans. The specific process is as follows:

[0141] Scanning electron microscopy (SEM) and transmission electron microscopy (TEM) were used to observe the structural disruption effect of G3 on Candida albicans: After Candida albicans was cultured in YPD medium for 24 hours, the bacterial concentration was adjusted to 2×10⁻⁶. 5 CFU / mL, and incubated with G3 (10 μM) for 30 minutes, followed by white light irradiation (0.06 W / cm²). 2 Alternatively, the samples were treated in the dark. Subsequently, the fungi were collected by centrifugation at 4000 rpm for 10 minutes and fixed overnight at 4°C with 2.5% glutaraldehyde, followed by fixation with 1% osmium tetroxide for 1–2 hours. The samples were then dehydrated sequentially with different concentrations of ethanol (30%, 50%, 70%, 80%, 90%, 95%, and 100%), maintaining each concentration for 15 minutes. Next, they were treated with a 1:1 mixture of ethanol and isoamyl alcohol for 30 minutes, followed by treatment with pure isoamyl alcohol for 1 hour. Finally, critical point drying and gold plating were performed, and the samples were observed using a scanning electron microscope (SEM, HITACHI SU8010). For transmission electron microscopy (TEM) samples, after gradient dehydration with ethanol, they were treated with pure acetone for 20 minutes, followed by treatment with a 1:1 (v / v) mixture of embedding agent and acetone for 1 hour, and then with a 3:1 (v / v) mixture of embedding agent and acetone for 3 hours. Next, the sections were treated with pure embedding medium and heated overnight at 70°C. Finally, ultrathin sections of 70–90 nm were prepared using an ultramicrotome (LEICA EMUC7). After staining and drying, the sections were observed using a transmission electron microscope (TEM, HITACHI H-7650). Figure 6 As shown in Figures b and c, scanning electron microscopy images reveal the collapse and rupture of the fungal surface after G3 treatment, while transmission electron microscopy images show blurred fungal cell contents and disruption of the cell wall and cell membrane. These results demonstrate that G3 can induce fungal death by disrupting internal and membrane structures, further confirming the fungicidal effect of the complex and providing a new treatment option for clinical fungal infections.

[0142] Example 10

[0143] This embodiment utilizes complexes G1-G5 for in vitro imaging and antifungal effects (drug-resistant strains). The specific process is as follows:

[0144] Five clinically isolated drug-resistant Candida species (Candida albicans, Candida krusei, Candida glabrata, Candida tropicalis, and Candida parapsilosis) were collected. All strains were provided by Nanfang Hospital, Southern Medical University. Fungal imaging and antifungal performance testing were performed according to the methods in Examples 6 and 8. Results are as follows: Figure 7 As shown, complex G3 also exhibits good imaging and antifungal effects against clinically isolated drug-resistant strains.

[0145] Experimental Example 2

[0146] This experiment tested the cytotoxicity of complexes G1–G5. The specific procedure was as follows:

[0147] MTT assay for cell viability: HaCaT and L929 cells were used to evaluate the in vitro cytotoxicity of G1–G5 cells using the MTT assay. Cells were cultured in DMEM medium containing 10% fetal bovine serum (FBS). HaCaT and L929 cells were sputtered at a rate of 1 × 10⁻⁶ cells / mL. 4 Cells were seeded at a density of 100 μL / well in 96-well plates and cultured at 37°C and 5% CO2 for 24 hours. Different concentrations of G1–G5 (0–15 μM) were added, and the culture was continued for another 24 hours. The medium was then replaced with fresh medium containing 0.5 mg / mL MTT. After incubation for 4 hours, the supernatant was discarded, and 150 μL of DMSO was added to each well to dissolve formazan crystals. The absorbance at 570 nm was measured using a microplate reader (PerkinElmer Victor3). Cell viability was calculated using the formula: Cell viability (%) = (A-treated group - A-blank) / (A-untreated group - A-blank) × 100%. As shown in Figure 8, the effects of complexes G1, G3, and G4 on normal cell viability at concentrations with antibacterial activity were negligible (cell viability was greater than 80% at a complex concentration of 7.5 μM). This demonstrates that complexes G1, G3, and G4 have good biocompatibility, further supporting their potential as selective antifungal agents.

[0148] Example 11

[0149] This embodiment uses lipidomics analysis to investigate the mechanism by which complex G3 induces fungal death. The specific process is as follows:

[0150] Candida albicans was cultured in YPD medium at 37°C for 24 hours, and the bacterial cells were collected and resuspended in PBS to a concentration of 5 × 10⁻⁶. 6 CFU / mL. Take 1 mL of bacterial suspension and incubate with G3 (5 μM) at 37°C for 30 minutes, then add 0.06 W / cm³. 2Irradiated with white light for 30 minutes, with the untreated group serving as a control. After treatment, bacterial cells were collected by centrifugation (4,000×g, 5 minutes, 4℃) and flash-frozen in liquid nitrogen. Lipidomics analysis was performed by Shanghai MajorBio Biotechnology Co., Ltd. Samples were analyzed using a Thermo UHPLC-Q ExactiveHF-XVanquish Horizon system with an Accucore C30 column (100mm×2.1mm inner diameter, 2.6μm; Thermo, USA). Raw data were used for lipid identification and quantification using LipidSearch (Thermo, USA) software, and subsequent analyses were performed on the MajorBio cloud platform (cloud.majorbio.com). Differentially metabolized metabolites were screened based on the variable importance projection (VIP) value and t-test p-value using the OPLS-DA model. Metabolites with VIP>1, p<0.05, and a fold change>2 were identified as significantly differentially metabolites. Results are as follows: Figure 9 As shown in Figures a and b, OPLS-DA analysis revealed significant changes in lipids in *Candida albicans* before and after G3 treatment. The analysis identified over 680 different lipids, including 382 glycerophospholipids (GP), 146 glycerolipids (GL), 133 sphingolipids (SP), 15 fatty acids (FA), and 4 sterol lipids (ST). Figure 9 (c) Further classification and statistical analysis. Further classification and statistical analysis based on the degree of lipid chain unsaturation showed 433 saturated fatty acids (SFA), 458 monounsaturated fatty acids (MUFA), 415 polyunsaturated fatty acids (PUFA), and 218 odd-numbered fatty acid acyl groups (ODD), the majority of which were polyunsaturated fatty acids. Figure 9 (d, e). After G3 treatment of fungi, 36 lipids showed significant changes, of which 12 lipids decreased and 24 lipids increased. Figure 9 The top 20 lipids with the greatest differences in abundance were identified in the sample, with phosphatidic acid (PA) showing the most significant variation. PA, as a precursor to phospholipids, participates in biomembrane formation; a decrease in PA can weaken the structural stability of cell membranes. Among the lipids with significant increases, most were PUFAs, whose accumulation is a key substrate for lipid peroxidation and ferroptosis. These findings lay the foundation for understanding G3-induced fungal ferroptosis.

[0151] Example 12

[0152] This embodiment studies the ferroptosis induced by complex G3 through TrxR activity inhibition in fungi. The specific process is as follows:

[0153] Culture Candida albicans overnight to the logarithmic growth phase, then adjust the concentration to 1×10⁻⁶ with PBS. 6CFU / mL. Take 500 μL of cell suspension and pre-incubate with the following inhibitors at 37℃ for 30 minutes: (1) Ferraphobia inhibitors: Liproxstatin-1 (Lip-1) and deferoxamine (DFX); (2) Necrostatin-1 (Nec-1) inhibitor; (3) Autophagy inhibitor: 3-methyladenine (3-MA); (4) Apoptosis inhibitor: Z-VAD-FMK. After inhibitor pretreatment, add G3 (5 μM) and continue incubation for 30 minutes. After the reaction is terminated, serially dilute with sterile PBS, take 20 μL and spread it on YPD agar plates. After incubation at 37℃ for 24 hours, take pictures of the plates and count the colonies using ImageJ software with the Colony Counter plugin. The total ROS and lipid ROS levels in G3-treated (white light irradiation) Candida albicans cells were detected by fluorescent probes DCFH-DA and C11-BODIPY630 / 650, respectively. The concentration of the Candida albicans suspension was adjusted to 1×10⁻⁶. 7 CFU / mL, divided into four treatment groups: (1) PBS; (2) PBS + light (0.06W / cm²). 2 (3) G3 (10μM); (4) G3 (10μM) + light (0.06W / cm²) 2 After treatment, the bacterial cells were collected by centrifugation at 4000g for 5 minutes and stained with DCFH-DA (20μM) and BODIPY 665 / 676 (10μM) for 30 minutes under dark conditions, followed by washing three times with PBS. Imaging was performed using a confocal microscope (CLSM, LSM880, Carl Zeiss, Germany). DCFH-DA: λ ex =488nm, λ em =490~520nm; BODIPY 665 / 676: Oxidation state (λ) ex =588nm, λ em =590~630nm), reduced state (λ) ex =633nm, λ em =630~670nm); Total glutathione peroxidase (GPX) activity was determined using the NADPH method Total glutathione peroxidase assay kit (Beyotime, China), and thioredoxin reductase (TrxR) activity was determined using the thioredoxin reductase activity assay kit (Beijing Bio-Sens Biotechnology Co., Ltd.). Results are as follows: Figure 10 As shown in Figure a, treatment with only the ferroptosis inhibitors Lip-1 and DFX significantly inhibited the antifungal activity of G3, indicating that ferroptosis may occur in Candida albicans after G3 treatment. This conclusion is further supported by increased lipid peroxidation levels (…). Figure 10 Changes in glutathione peroxidase (Gpx) levels (b, c) Figure 10 Supported by d). Furthermore, compound G3 inhibited the activity of thioredoxin reductase (TrxR) in Candida albicans in a dose-dependent manner. Figure 11 (a) Given the crucial role of TrxR in the antioxidant defense system, its inhibition would disrupt cellular antioxidant defense mechanisms, leading to increased levels of reactive oxygen species (ROS). Consistent with this, significant ROS accumulation was observed after G3 treatment. Figure 10 e, Figure 11 (b) To further verify the regulatory role of TrxR in ferroptosis, cell viability and lipid peroxidation levels were assessed after TrxR was inhibited by TRi-1. Figure 11 (c-e). Experimental results showed that TRi-1 treatment led to a decrease in the survival rate of Candida albicans and the accumulation of lipid peroxidation products. These results confirm that G3 can regulate the ferroptosis process in Candida albicans by inhibiting TrxR activity.

[0154] Example 13

[0155] This embodiment uses GSEA analysis to investigate mitochondrial dysfunction driven by complex G3. The specific procedure is as follows:

[0156] The sample pretreatment steps are the same as the aforementioned SEM characterization method: samples treated with G3 (5 μM) combined with white light irradiation (0.06 W / cm²) were then processed. 2 Candida albicans cells were collected by centrifugation (4,000×g, 15 min) from treated (30 min) and untreated (control) cells, washed with PBS, and then flash-frozen in liquid nitrogen. RNA sequencing was performed by Shanghai MajorBio Technology Co., Ltd. Sequencing was performed using the Illumina HiSeq4000 sequencing platform. Data were analyzed online through the MajorBio cloud platform (cloud.majorbio.com). Raw reads were filtered to obtain high-quality data. Differentially expressed genes (fold change|>2, P<0.05) were screened and pathway enrichment analysis was performed. Results are as follows: Figure 12 As shown in a and b, GSEA analysis revealed that G3, after being located in the mitochondria, disrupts pathways related to mitochondrial function, including the tricarboxylic acid cycle and oxidative phosphorylation.

[0157] Example 14

[0158] This embodiment investigates the effects of complex G3 on fungal mitochondrial dysfunction and oxidative stress. The specific process is as follows:

[0159] Culture Candida albicans overnight to the logarithmic growth phase, then adjust the concentration to 5 × 10⁻⁶ with PBS. 6CFU / mL (MCF = 5). 500 μL of cell suspension was subjected to four treatments: (1) PBS; (2) PBS + white light irradiation (0.06 W / cm²). 2 (3) G3 (10μM); (4) G3 (10μM) + white light irradiation (0.06W / cm²) for 30 minutes; (5) G3 (10μM) + white light irradiation (0.06W / cm²) for 30 minutes; (6) G3 (10μM) + white light irradiation (0.06W / cm²) for 30 minutes; (7) G3 (10μM); (8) 2 After treatment, cells were collected by centrifugation at 4000×g for 5 minutes and washed twice with PBS. Staining was performed using the JC-1 fluorescent probe (10 μg / mL) at 37℃ in the dark for 30 minutes. After final centrifugation, the stained cells were resuspended in PBS and immediately analyzed using a confocal laser scanning microscope (CLSM, LSM880, Carl Zeiss, Germany). Aggregate detection parameters: excitation wavelength 488 nm, emission wavelength 550-595 nm; monomer detection parameters: excitation wavelength 488 nm, emission wavelength 500-550 nm. The fluorescence index was expressed as the ratio of red-green fluorescence density to the control group. ATP content was determined using an enhanced ATP assay kit (Beyotime, China), following the manufacturer's instructions. The mitochondrial ROS level in G3-treated (white light irradiated) Candida albicans cells was detected using the MitoSOX fluorescent probe. The concentration of the Candida albicans suspension was adjusted to 1×10⁻⁶. 7 CFU / mL, divided into four treatment groups: (1) PBS; (2) PBS + light (0.06W / cm²). 2 (3) G3 (10μM); (4) G3 (10μM) + light (0.06W / cm²) 2 After treatment, the cells were collected by centrifugation at 4000g for 5 minutes, stained with MitoSOX (2μM) for 30 minutes in the dark, and washed three times with PBS. Imaging was performed using a confocal microscope (CLSM, LSM880, Carl Zeiss, Germany). MitoSOX: λ ex =590nm, λ em =600~630nm. Results are as follows: Figure 12 As shown in c and d, G3 leads to a significant decrease in mitochondrial membrane potential and ATP levels, indicating mitochondrial dysfunction. Figure 12 The presence of e indicates an increase in ROS in mitochondria, suggesting oxidative stress in the mitochondria.

[0160] Example 15

[0161] This embodiment studies the effects of complex G3 on fungal mitochondrial dysfunction and oxidative stress-driven ferroptosis. The specific process is as follows:

[0162] As described in Examples 12 and 8, lipid peroxidation levels and fungal survival rates in *Candida albicans* were detected after treatment with mito-tempo. The results are as follows: Figure 12As shown in f and g, after using the mitochondrial reactive oxygen species scavenger Mito-TEMPO, the increased lipid peroxidation level and antifungal effect in G3-treated Candida albicans were reversed to some extent, indicating that G3 drives ferroptosis in Candida albicans by inducing mitochondrial dysfunction and oxidative stress.

[0163] Example 16

[0164] This embodiment studies the effect of complex G3 on the fungal mitochondrial-iron cascade reaction, and the specific process is as follows:

[0165] Transcriptome sequencing was performed as described in Example 13, and differentially expressed genes were analyzed for GO enrichment. Candida albicans was cultured overnight to the logarithmic growth phase, and the concentration was adjusted to 5 × 10⁻⁶ PBS. 6 CFU / mL. Take 500 μL of cell suspension and perform the following four treatments respectively: (1) PBS; (2) PBS + white light irradiation (0.06 W / cm²). 2 (3) G3 (10μM); (4) G3 (10μM) + white light irradiation (0.06W / cm²) for 30 minutes; (5) G3 (10μM) + white light irradiation (0.06W / cm²) for 30 minutes; (6) G3 (10μM) + white light irradiation (0.06W / cm²) for 30 minutes; (7) G3 (10μM); (8) 2 After treatment, cells were collected by centrifugation at 4000g for 5 minutes, washed twice with PBS, and incubated at 37°C in the dark for 30 minutes with 10 μg / mL FerroOrange fluorescent probe (Dongren Chemical Research Institute). After staining, cells were centrifuged again (4000g, 5 minutes), resuspended in PBS, and immediately imaged using a confocal laser scanning microscope (CLSM, LSM880, Carl Zeiss, Germany) with excitation wavelength of 543 nm and emission wavelength of 550–600 nm. Figure 13 As shown in Figure a, GO enrichment analysis indicated that G3 treatment significantly altered the iron ion homeostasis of fungi, and G3 significantly increased the iron ion level in Candida albicans. Figure 13 (b) Mitochondria, as the central hub of iron metabolism, maintain iron homeostasis by coordinating the synthesis and storage of iron-sulfur clusters and heme. Further gene expression profiling revealed that G3 treatment significantly upregulated genes related to fungal iron transport, iron reduction, iron-sulfur cluster synthesis and assembly, and heme binding. Figure 13 (c~f).

[0166] Example 17

[0167] This embodiment studies the in vivo antifungal effect of complex G3. The specific process is as follows:

[0168] Six-week-old female BALB / c mice (weighing 18–22 g) were anesthetized via intraperitoneal injection of sodium pentobarbital (75 mg / kg body weight). The fur on the backs of the mice was removed using a shaver, and after disinfection with alcohol, full-thickness skin defects were created on the backs using a scalpel. Each wound was inoculated with 10 μL of Candida albicans suspension (1 × 10⁻⁶ mg / kg body weight). 8 After drying, the wound was covered with sterile breathable tape. 24 hours after infection, the mice were randomly divided into 5 treatment groups (4 mice per group): (1) PBS control group (50 μL); (2) PBS + light group (50 μL, 0.06 W / cm²). 2 (3) White light irradiation for 30 min); (4) Voriconazole group (20 μM, 50 μL); (5) G3 group (20 μM, 50 μL); (6) G3+ light irradiation group (20 μM, 50 μL, 0.06 W / cm²). 2 White light irradiation for 30 minutes). The treatment method involved direct application to the wound, repeated every 24 hours for 3 consecutive days. Body weight and wound area were monitored daily. On day 11, blood samples, wound tissue, and organs were collected from mice, and the mice were euthanized. Wound tissue was stained with hematoxylin and eosin (H&E), Masson staining, and immunohistochemically analyzed for IL-10, IL-6, and CD31 (to assess angiogenesis). Quantitative analysis was performed using ImageJ. Organs (heart, liver, spleen, lung, kidney) were stained with H&E to evaluate the biocompatibility of G3. Results are as follows: Figure 14 As shown in a and b, compared with the PBS solution-treated control group, the voriconazole, G3, and G3+ light-treated groups significantly promoted the healing of mouse skin wounds. H&E and Masson staining showed that G3+ light treatment significantly reduced inflammatory cell infiltration, accelerated granulation tissue formation, and promoted collagen deposition. Figure 14 (c, d, h, i). Furthermore, G3+ light treatment significantly increased the expression of pro-inflammatory factors IL-6 and TNF-α (c, d, h, i). Figure 14 (e, f, j, k). Meanwhile, G3+ phototherapy significantly promoted angiogenesis, which is beneficial for wound repair and healing. Figure 14 These results indicate that G3 has the effect of controlling fungal infection of wounds and promoting wound healing, providing a new treatment option for fungal wound infections in clinical practice. Furthermore, H&E staining of major organs (heart, liver, spleen, lung, and kidney) showed no obvious pathological damage after different treatments, and there were no differences in morphology and structure. Figure 15 (a) Liver and kidney function indicators (ALT, AST, CREA, BUN) Figure 15 All values ​​(b-e) were within the normal range. During the treatment period, the body weight of the mice in each group did not change significantly. Figure 15(f). These results indicate that G3 has good biocompatibility and safety, further supporting its safety in wound infection control. This invention can provide a new treatment option for clinical drug-resistant fungal skin wound infections.

[0169] Example 18

[0170] This embodiment uses a hemolysis test to study the in vivo biocompatibility of complex G3. The specific process is as follows:

[0171] Fresh mouse blood was collected and centrifuged at 1500 rpm for 15 minutes. The supernatant was discarded, and the red blood cells were washed three times with physiological saline. The red blood cells were then diluted 20-fold with physiological saline to prepare a working suspension. The red blood cell suspension was mixed with equal volumes of different concentrations (0-50 μM) of G3 compound and incubated at 37°C for 24 hours. After incubation, the supernatant was collected by centrifugation at 1500 rpm for 15 minutes. A negative control (physiological saline treatment group) and a positive control (distilled water treatment group) were also included in the experiment. The absorbance of each group at 405 nm was measured using a microplate reader, and the hemolysis rate was calculated using the following formula: Hemolysis rate (%) = [(Absorbance of G3 treatment group - Absorbance of negative control group) / (Absorbance of positive control group - Absorbance of negative control group)] × 100%. Results are as follows: Figure 15 As shown in g and h, G3 treatment of erythrocytes did not cause significant hemolysis (the hemolysis rate was less than 13% when the complex concentration was 50 μM), indicating its biocompatibility in vivo.

[0172] Example 19

[0173] This embodiment uses a rat corneal fungal infection model to study the in vivo antifungal effect of complex G3. The specific process is as follows:

[0174] Female Sprague-Dawley rats (8 weeks old, 150–170 g) were anesthetized by intraperitoneal injection of 1% sodium pentobarbital, and their corneal epithelium was abraded. 10 μL of Candida albicans suspension (1 × 10⁻⁶) was then added. 8 A fungal keratitis model was established by injecting CFU / ml into the superficial corneal stroma. After 48 hours, patients were randomly divided into 5 treatment groups (n=4) according to the previous wound infection model: (1) PBS control group (50 μL); (2) PBS + light irradiation group (50 μL, 0.06 W / cm²). 2 (3) White light irradiation for 30 min); (4) Voriconazole group (20 μM, 50 μL); (5) G3 group (20 μM, 50 μL); (6) G3+ light irradiation group (20 μM, 50 μL, 0.06 W / cm²). 2White light irradiation for 30 minutes). Treatment involved direct application to the corneal surface three times daily. The cornea was observed and photographed every two days. Clinical severity was assessed by three masked observers based on edema, surface regularity, neovascularization, opacity area, and opacity density (each score ranged from 0-3, total from 0-15). On day 7, corneal epithelial defects were assessed using 2 μL of 0.5% sodium fluorescein. On day 10, rats were euthanized, and corneas were harvested, rinsed with PBS, homogenized, and spread onto YPD plates for incubation at 37°C for 24 hours, followed by CFU counting. Corneal tissue was also collected for histological analysis. Furthermore, the material was used to treat the corneas of healthy mice, and H&E staining was used to evaluate its biocompatibility. After three days of continuous treatment, the control group still showed corneal opacity and edema, the G3 and voriconazole groups showed moderate treatment effects, while the G3+ light irradiation group showed significant reduction in corneal opacity and edema. Figure 16 (a) The lowest clinical score ( Figure 16 (d) Fluorescein staining showed that the corneal epithelial damage was minimal in the G3+ phototherapy group (lesion area was 0.32), while the lesion areas in the control group, phototherapy group, voriconazole group, and G3 group were 1, 0.96, 0.65, and 0.45, respectively. Figure 16 (b, e). The lowest number of fungal colonies was observed, with a relative fungal load as low as 0.12 (the fungal loads in the control group, light group, voriconazole group, and G3 group were 1, 0.99, 0.41, and 0.25, respectively). Figure 16 The treatment effect was best in the C and F groups. H&E staining showed that the control group had abnormal corneal structure and more inflammatory cells, while the G3+ light irradiation group had significantly reduced corneal infiltration, corneal thinning, and reduced swelling. Figure 16 (g). The results demonstrate the significant potential and advantages of G3 in treating fungal keratitis. This invention can provide a new treatment option for clinical drug-resistant fungal corneal infections.

[0175] Example 20

[0176] This embodiment uses a systemic fungal infection model to study the in vivo antifungal effect of complex G3. The specific process is as follows:

[0177] Six-week-old female ICR mice (20–22 g) were immunosuppressed by intraperitoneal injection of cyclophosphamide (200 mg / kg) three days before infection. Mice were then intravenously injected with 100 μL of Candida albicans (5 × 10⁻⁶ g / kg). 6 Infection model was established using CFU / ml. After 24 hours, the skin wound infection model was randomly divided into 5 groups (n=4): (1) PBS control group (50μL); (2) PBS + light irradiation group (50μL, 0.06W / cm²). 2(3) White light irradiation for 30 min); (4) Voriconazole group (500 μM, 50 μL); (5) G3 group (500 μM, 50 μL); (6) G3+ light irradiation group (500 μM, 50 μL, 0.06 W / cm²). 2 White light irradiation for 30 minutes). G3 (500 μM) was administered intravenously every 24 hours for 6 consecutive days. PBS and voriconazole (500 μM) served as negative and positive controls, respectively. Mice mortality was monitored during treatment, and major organs (heart, liver, spleen, lung, kidney) from dead mice were collected, homogenized in PBS, and plated on YPD agar for CFU counting. Surviving mice were euthanized after completing the treatment protocol, and their organs were collected, fixed overnight in 4% paraformaldehyde, embedded in paraffin, and sectioned at 5 μm thickness for H&E staining. Results are as follows. Figure 17 In the control group (A), the survival rate was low, with all mice dying within 4 days of infection. Treatment with G3 and voriconazole significantly prolonged the survival time and improved the survival rate of infected mice. Simultaneously, G3 and G3+ light exposure reduced the fungal load in mouse organs. Figure 17 (b-f). H&E staining showed that G3 effectively alleviated typical pathological changes, such as inflammatory infiltration and necrosis, especially in the kidneys ( Figure 17 The results (g) showed that G3 had good in vivo therapeutic effects. This invention can provide a new treatment option for systemic infections caused by drug-resistant fungi in clinical practice.

[0178] The above embodiments are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above embodiments. Any changes, modifications, substitutions, combinations, or simplifications made without departing from the spirit and principle of the present invention shall be considered equivalent substitutions and shall be included within the protection scope of the present invention.

Claims

1. Compounds of Formula I, Formula II, or pharmaceutically acceptable salts thereof: ； ； in, R1, each of R2, and each of R4 are each independently selected from H, halogens, C1-C6 alkyl groups, and C1-C6 alkoxy groups; the alkyl and alkoxy groups are optionally unsubstituted or substituted by one or more R groups. a replace; Each R3 is independently selected from halogens, C6~C6. 20 Aryl, C6~C 20 arylphosphinyl; the aryl or arylphosphinyl group may optionally be unsubstituted or substituted with one or more R groups. a replace; Each R a Selected from halogens and C1-C6 alkyl groups.

2. The compound according to claim 1, characterized in that: Each R3 is independently selected from F, Cl, Br, I, benzene ring, biphenyl, diphenylphosphinyl, and triphenylphosphinyl; Each phenyl group is optionally unsubstituted, or substituted with one or more R groups. a replace.

3. The compound according to claim 1, characterized in that: The compound is selected from the following compounds: 、 、 、 、 。 4. A method for preparing the compound according to any one of claims 1 to 3, characterized in that: Includes the following steps: Compound I-1 and compound I-2 were subjected to a Suzuki coupling reaction, then reacted with imidazole, followed by a quaternization reaction with compound I-3, and then an exchange reaction to obtain compound I. 、 、 ； Compound II-1 and compound II-2 were subjected to a Suzuki coupling reaction, followed by reaction with imidazole, then quaternization with compound I-3, and finally exchange reaction to obtain compound II. 、 ； Wherein, R1, R2, and R4 are defined as described in any one of claims 1 to 3; R5 and R6 are each independently selected from any one of borate group, trifluoroborate group, triol borate group, and borate ester group; X, X1, X2, X3, and X4 are each independently selected from halogens.

5. A pharmaceutical composition comprising the compound according to any one of claims 1 to 3, and optionally, pharmaceutically acceptable excipients.

6. The pharmaceutical composition according to claim 5, characterized in that: The pharmaceutical composition is a fungal fluorescence imaging agent.

7. The use of the compound according to any one of claims 1 to 3 or the pharmaceutical composition according to claim 5 or 6 in the preparation of drugs for the prevention and / or treatment of fungal infections or diseases, or fluorescent probes.

8. The application according to claim 7, characterized in that: The fluorescent probe is used for fungal fluorescence imaging.

9. The application according to claim 7, characterized in that: Fungal infections to be prevented and / or treated are selected from candidemia, invasive candidiasis, tinea capitis, tinea corporis, tinea pedis, onychomycosis, periungual candidiasis, tinea versicolor, oral thrush, fungal keratitis, vaginal candidiasis, respiratory candidiasis, biliary candidiasis, esophageal candidiasis, urinary candidiasis, systemic candidiasis, cutaneous and mucosal candidiasis, aspergillosis, mucosomycosis, South American blastomycosis, North American blastomycosis, histoplasmosis, coccidioidomycosis, sporotrichosis, or chronic sinusitis.

10. The application according to claim 9, characterized in that: The chronic sinusitis mentioned includes fungal sinusitis.

11. The application according to claim 7, characterized in that: The drug is used in photodynamic therapy, wherein the light conditions for photodynamic therapy are: light intensity 0.01~2.0 W / cm². 2 The illumination time is 10~60min.

12. The application according to claim 11, characterized in that: In the photodynamic therapy, the light conditions are: the light source is white light of 300~700nm.

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