Gold complex with aggregation-induced emission characteristic and preparation method and application thereof

Abstract

The invention discloses a gold complex with an aggregation-induced emission characteristic and a preparation method and application thereof, and provides a compound shown as a formula I, a compound shown as a formula II or pharmaceutically acceptable salts thereof. And a fluorescent dye with excellent performance is provided for fungal cell structure dyeing. The compound disclosed by the invention can drive fungal ferroptosis by inducing mitochondrial dysfunction and oxidative stress, so that multimodal mechanism activated ferroptosis such as genes related to fungal iron transport, iron reduction, iron-sulfur cluster synthesis and assembly and heme binding is remarkably up-regulated, and a scheme with a long-term curative effect is provided for clinical prevention and treatment of fungal drug-resistant strains.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of biological medicine, in particular to a gold complex with aggregation-induced emission characteristics and a preparation method and application thereof. BACKGROUND

[0002] Ferroptosis is a regulated form of cell death characterized by iron homeostasis imbalance, lethal accumulation of reactive oxygen species (ROS), depletion of oxidative stress defense systems, and abnormal lipid peroxidation. Increasing evidence suggests that ferroptosis plays a key regulatory role in many diseases, including neurodegenerative diseases (Alzheimer's disease, Parkinson's disease, and Huntington's disease), cancer development, stroke, cerebral hemorrhage, ischemia-reperfusion injury, traumatic brain injury, and kidney degeneration, making it a promising target for developing new therapeutic strategies. The molecular mechanisms of ferroptosis in mammalian cells have been well established, and the development of ferroptosis inhibitors or inducers has become a focus in the field of disease treatment, particularly in the field of anti-tumor therapy. Notably, iron-activated ferroptosis-like cell death has been observed in bacteria. However, research on ferroptosis in the field of antifungal agents is still significantly insufficient, and there are very limited reports on this topic. In particular, it is currently unclear whether ferroptosis can serve as a target for treating fungal infections.

[0003] In recent years, monovalent gold complexes have received attention as effective inducers of ferroptosis. Their mechanism of action involves targeting the inhibition of thioredoxin reductase (TrxR) through covalent binding to selenium-sulfur groups (such as Sec498), leading to irreversible inactivation of the enzyme. TrxR, as a key antioxidant enzyme, maintains the redox homeostasis within cells and regulates the antioxidant defense system by reducing thioredoxin (Trx). Inhibition of TrxR impairs ROS clearance, triggering a series of events, including collapse of the antioxidant system, iron metabolism disorder, impaired repair of lipid peroxidation, and oxidative membrane damage. These findings highlight monovalent gold complexes as a new strategy for inducing ferroptosis and overcoming tumor drug resistance. However, unlike tumor cells, fungal cells can develop resistance through genetic mutations, metabolic reprogramming, or enhanced antioxidant defenses, which may undermine the long-term efficacy of single-pathway ferroptosis inducers. Therefore, developing new drugs capable of activating ferroptosis through multi-modal mechanisms has important clinical significance. SUMMARY

[0004] The present application aims to solve at least one of the above technical problems in the prior art. To this end, the purpose of the present application is to provide a gold complex with aggregation-induced emission characteristics and a preparation method and application thereof.

[0005] To achieve the above-mentioned purpose, the technical solution adopted by the present application is:

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

[0007]

[0008]

[0009] wherein R1, each R2, and each R4 are each independently selected from H, halogen, C1-C6 alkyl, C1-C6 alkoxy; said alkyl, alkoxy is optionally unsubstituted, or substituted with one or more R a substituents;

[0010] each R3 is independently selected from halogen, C6-C10 aryl, C6-C10 aryl phosphine; said aryl, aryl phosphine is optionally unsubstituted, or substituted with one or more R 20 substituents; 20 substituents; a

[0011] each R a is selected from halogen, C1-C6 alkyl.

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

[0013] In some embodiments of the present application, the C1-C6 alkyl 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, 3,3-dimethyl-2-butyl.

[0014] In some embodiments of the present application, the C1-C6 alkoxy is selected from methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, sec-butoxy, isobutoxy, t-butoxy, n-pentoxy, neopentoxy, isopentoxy, t-pentoxy, n-hexoxy, cyclohexoxy, neohexoxy.

[0015] In some embodiments of the present application, the C6-C10 aryl is selected from phenyl, naphthyl, anthryl. 20 In some embodiments of the present application, the C6-C10 aryl is selected from phenyl, naphthyl, anthryl.

[0016] In some embodiments of the present application, the C6-C10 aryl is selected from phenyl, naphthyl, anthryl. 20 In some embodiments of the present application, the C6-C10 aryl phosphine is selected from diphenyl phosphine, triphenyl phosphine.

[0017] ​In some embodiments of the present application, each R3is independently selected from F, Cl, Br, I, phenyl ring, biphenyl, diphenylphosphino, triphenylphosphino; wherein each phenyl is optionally unsubstituted, or substituted with one or more R a substituted.

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

[0019]

[0020]

[0021] In a second aspect of the present application, a method for preparing the compound is provided, comprising the following steps:

[0022] The compound of formula I is prepared by subjecting a compound of formula I-1 to Suzuki coupling reaction with a compound of formula I-2, followed by reaction with imidazole, then quaternary amination reaction with a compound of formula I-3, and then exchange reaction.

[0023]

[0024] The compound of formula II is prepared by subjecting a compound of formula II-1 to Suzuki coupling reaction with a compound of formula II-2, followed by reaction with imidazole, then quaternary amination reaction with a compound of formula I-3, and then exchange reaction.

[0025]

[0026] wherein R1, R2, R4are as defined above; R5, R6are each independently selected from any one of boronic acid group, triflate group, triol borate group, borate ester group; X, X1, X2, X3, X4are each independently selected from halogen.

[0027] In some embodiments of the present application, as boronic acid group, borate ester group, triflate group or triol borate group, there is no particular limitation, for example, the following groups shown in the structural formula can be exemplified:

[0028]

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

[0030] In some embodiments of the present application, the palladium catalyst is selected from at least one of palladium tetra-triphenylphosphine, palladium acetate and 1,1'-bis(diphenylphosphino)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 application, 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 application, 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, the volume ratio of the organic solvent to water is (3-10):1, preferably 5:1; 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 application, the Suzuki coupling reaction is carried out under reflux conditions; the Suzuki coupling reaction is carried out for 1-8 h at a temperature of 70-120°C.

[0034] In some embodiments of the present application, the reaction of the Suzuki coupling reaction product with imidazole is a C-N coupling reaction; the C-N coupling reaction is carried out in the presence of a copper catalyst and a base; the copper catalyst comprises at least one of copper acetate, copper trifluoroacetate, copper triflate, copper acetylacetonate, copper sulfate, copper chloride, copper bromide, copper iodide, cuprous oxide, cuprous bromide, cuprous iodide, 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 C-N 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-volume ratio of the Suzuki coupling reaction product to the solvent is 1:(50-80), preferably 1:(55-65); the reaction temperature for the C-N coupling reaction is 130-180°C, such as 140-180°C; the reaction time is 12-48 h.

[0035] In some embodiments of the present application, the reaction temperature of the quaternary amination reaction is 80-120°C, such as 90-110°C; the reaction time is 10-24h.

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

[0037] In some embodiments of the present application, the anion exchange reaction is carried out using a silver salt, which includes at least one of Ag2O, Ag2CO3, AgOAc, Ag2S. The anion exchange reaction is carried out in the dark; the reaction time is 24-72h.

[0038] In some embodiments of the present application, 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, (IMes)AuC6F5. The reaction time of the ligand exchange reaction is 10-24h.

[0039] In a third aspect of the present application, a pharmaceutical composition is provided, which comprises the compound described above, and optionally, a pharmaceutically acceptable excipient.

[0040] The pharmaceutical composition of the present application is suitable for various administration routes, and thus can be prepared into any dosage form which is pharmaceutically acceptable. For example, the pharmaceutical composition described above can be administered to a patient or subject in need of such treatment by oral, parenteral, rectal or pulmonary administration, etc. When used for oral administration, the pharmaceutical composition can be prepared into oral preparations, such as conventional oral solid preparations, e.g. tablets, capsules, pills, granules, etc.; or oral liquid preparations, e.g. oral solutions, oral suspensions, syrups, etc. When prepared into oral preparations, suitable fillers, binders, disintegrants, lubricants, etc. can be added. When used for parenteral administration, the pharmaceutical composition described above can also be prepared into injections, including injection solutions, sterile powders for injection and concentrated solutions for injection. When prepared into injections, conventional methods in the existing pharmaceutical field can be used for production, and no additional agents can be added or suitable additional agents can be added according to the nature of the drug when the injection is prepared. When used for rectal administration, the pharmaceutical composition can be prepared into suppositories, etc. When used for pulmonary administration, the pharmaceutical composition can be prepared into inhalation preparations, aerosols, powder sprays or spray preparations, etc.

[0041] A pharmaceutically acceptable excipient refers to a substance that is nontoxic, compatible with the active ingredient, and otherwise biologically suitable for use in the body. The choice of a particular excipient will depend on the mode of administration or the type and state of the disease for which a particular patient is being treated. Examples of pharmaceutically acceptable excipients include, but are not limited to, solvents, diluents, dispersing agents, suspending agents, surfactants, isotonic agents, thickening agents, emulsifiers, binders, lubricants, stabilizers, hydrating agents, emulsification accelerators, buffers, absorbents, colorants, ion exchangers, release agents, coating agents, flavoring agents, and antioxidants, and the like, which are conventional in the pharmaceutical art. If necessary, flavoring agents, preservatives, and sweetening agents, and the like, can also be added to the pharmaceutical composition.

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

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

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

[0045] In a fourth aspect of the present application, there is provided a use of the compound in the preparation of a medicament for preventing and / or treating a fungal infection or disease, a fluorescent probe.

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

[0047] In some embodiments of the present application, the fungal infection or disease comprises a fungal susceptible strain infection or disease and / or a fungal drug-resistant strain infection or disease.

[0048] In some embodiments of the application, the fungal infection to be prevented and / or treated is selected from candidaemia, invasive candidiasis, tinea capitis, tinea corporis, tinea pedis, onychomycosis, perionychomycosis, tinea versicolor, oral thrush, fungal keratitis, vaginal candidiasis, respiratory candidiasis, biliary candidiasis, oesophageal candidiasis, urological candidiasis, systemic candidiasis, mucocutaneous candidiasis, aspergillosis, mucormycosis, South American blastomycosis, North American blastomycosis, histoplasmosis, coccidioidomycosis, sporotrichosis, mycetoma, fungal rhinosinusitis or chronic rhinosinusitis. In some embodiments, the infection is candidaemia or invasive candidiasis.

[0049] In some embodiments of the application, the fungal infection to be prevented and / or treated comprises an infection of at least one of Candida albicans, C. glabrata, C. dubliniensis, C. krusei, C. parapsilosis, C. tropicalis, C. orthopsilosis, C. guilliermondii, C. rugosa, C. auris, C. lusitaniae, Aspergillus fumigatus, A. flavus, A. terreus, A. niger, A. candidus, A. clavatus or A. ochraceus.

[0050] In some embodiments of the application, the fungal infection to be prevented and / or treated comprises an infection of at least one of C. albicans, C. krusei, C. glabrata, C. tropicalis and C. parapsilosis.

[0051] In some embodiments of the application, the drug induces fungal ferroptosis, drives fungal mitochondrial dysfunction (such as tricarboxylic acid cycle and / or oxidative phosphorylation dysfunction, reduced membrane potential and / or ATP levels, oxidative stress).

[0052] In some embodiments of the application, the drug induces fungal ferroptosis by inhibiting TrxR activity.

[0053] In some embodiments of the application, the drug is used in a photodynamic therapy, wherein the light exposure conditions are: light intensity 0.01-2.0 W / cm22 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 Experimental Example 1 of the present invention; b represents the ability of complexes G1 to G5 in Experimental 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 In vitro antifungal (standard strain) effect of complexes G1-G5 in Example 8 of the present application, a is a plate antibacterial result, b, c are scanning electron microscopy and transmission electron microscopy results, respectively.

[0064] Figure 7 In vitro imaging (a) and antifungal effect on drug-resistant strains (b) of complexes G1-G5 in Example 10 of the present application.

[0065] Figure 8 Toxicity test of complexes G1-G5 in Test Example 2 of the present application on HaCaT cells (a) and L929 cells (b).

[0066] Figure 9 Lipidomics analysis of the mechanism of complex G3-induced fungal death in Example 11 of the present application; a, b are OPLS-DA analysis of differential genes before and after G3 treatment; c-e are lipid species and lipid unsaturation analysis; f is differential lipid analysis.

[0067] Figure 10 Verification of complex G3-induced fungal ferroptosis by TrxR activity inhibition in Example 12 of the present application; a is the change in antibacterial effect after treatment with different cell death inhibitors; b, c are qualitative and quantitative analysis of lipid peroxidation levels in Candida albicans under different treatment conditions; d is Gpx activity level analysis in the fungus; e is qualitative analysis of reactive oxygen species levels in Candida albicans.

[0068] Figure 11 Verification of complex G3-induced fungal ferroptosis by TrxR activity inhibition in Example 12 of the present application; a is TrxR activity level analysis in the fungus; b is quantitative analysis of reactive oxygen species levels in Candida albicans; c-e are evaluation of cell viability and lipid peroxidation levels after TRi-1 inhibits TrxR, respectively.

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

[0070] Figure 13For the complex G3 in Example 16 of the present application to verify the fungal mitochondrial-iron ion cascade reaction, a is GO enrichment analysis; b is Candida albicans intracellular iron ion staining; c-f are the expression of genes related to fungal iron transport, iron reduction, iron-sulfur cluster synthesis and assembly, and hemoglobin binding, respectively.

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

[0072] Figure 15 For the biological safety evaluation of mice in Example 17 and Example 18 of the present application, a is the HE staining of the heart, liver, spleen, lung, and kidney tissues of mice; b-e are the blood biochemical results of mice; f is the body weight record of mice; g and h are the hemolysis experiment of complex G3.

[0073] Figure 16 For the treatment effect of the mouse fungal keratitis infection model in Example 19 of the present application; a and d represent the recovery of the mouse cornea and the clinical grade score; b and e represent the fluorescein sodium 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 For the treatment effect of the mouse systemic infection model in Example 20 of the present application; a is the survival rate curve of mice; b-f represent the fungal load of mouse organs; g is the HE staining of the main organs.

[0075] Legend: * indicates p<0.05; ** indicates p<0.005; *** indicates p<0.001. DETAILED DESCRIPTION

[0076] The content of the present application is further described in detail through specific examples. The raw materials, reagents or devices used in the examples and comparative examples can be obtained from conventional commercial channels, or can be obtained by existing technical methods, unless otherwise specified. Unless otherwise specified, the test or test method is a conventional method in the art.

[0077] The term "pharmaceutically acceptable" means chemically or physically compatible with the other ingredients constituting a pharmaceutical dosage form, and physiologically compatible with the recipient.

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

[0079] Example 1

[0080] This example prepared gold complex G1, the specific process is as follows:

[0081]

[0082] 1. Synthesis of compound 1c

[0083] Into a 100 mL reaction flask were added 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). To the reaction flask was then added a solvent system consisting of THF (50 mL) and H2O (10 mL). The reaction mixture was refluxed in an 80 °C oil bath, and the progress of the reaction was monitored by thin layer chromatography (TLC) until completion. After cooling to room temperature, the crude product was extracted and isolated, followed by purification through silica gel column chromatography to obtain the target compound 1c as an orange solid (1.05 g, yield 46%).

[0084] 2. Synthesis of compound 1e

[0085] Into a 100 mL reaction flask were added 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), and combined. To the reaction flask was then added DMF (50 mL), and the mixture was heated to reflux for 24 hours. After the reaction was completed, the mixture was cooled, and a large amount of ice water was added to precipitate 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 was used directly in the next reaction without further purification.

[0086] 3. Synthesis of compound 1f

[0087] To a 100 mL reaction flask was added 1e (2.23 g, 5.00 mmol). To the flask was then added iodomethane (1.42 g, 10.00 mmol) and DMF (50 mL). The mixture was refluxed at 100 °C for 12 h. After the reaction was completed, 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 red solid 1f (2.05 g, yield 70%). 1 H NMR (400 MHz, CDC13) δ 10.60 (s, 1H), 8.51 (d, J = 6.4 Hz, 1H), 8.37 (s, 1H), 7.92 (s, 1H), 7.82 (t, J = 6.0 Hz, 3H), 7.30 (t, J = 7.6 Hz, 4H), 7.17 (d, J = 8.4 Hz, 5H), 7.13 (s, 1H), 7.09 (t, J = 7.6 Hz, 2H), 4.37 (s, 3H). 13 C NMR (101 MHz, CDC13) δ 154.0, 149.1, 148.0, 147.0, 136.6, 136.0, 130.2, 129.5, 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] To a 50 mL reaction flask was added 1f (0.18 g, 0.30 mmol) and Ag20 (0.04 g, 0.15 mmol). To the flask was then introduced DCM (30 mL). The mixture was stirred at room temperature for 48 h in the dark. Then (htt)AuCl (0.10 g, 0.30 mmol) was added and stirring was continued for 12 h. The reaction mixture was filtered through celite and the filtrate was evaporated under reduced pressure. The crude product was purified by flash column chromatography on neutral alumina using DCM as eluent and then recrystallized from a DCM / hexane mixture to give the desired red solid G1 (0.15 g, yield 70%). 1 H NMR (400 MHz, CDC13) δ 8.23 (d, J = 7.6 Hz, 1H), 7.85 (d, J = 8.4 Hz, 2H), 7.78 (d, J = 7.6 Hz, 1H), 7.62 (s, 1H), 7.31 (t, J = 7.6 Hz, 4H), 7.20 (d, J = 8.0 Hz, 7H), 7.09 (t, J = 7.2 Hz, 2H), 4.02 (s, 3H). 13C NMR (101 MHz, CDC13) 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 spectrum: m / z: [M+Na] 714.095; found: 714.0757. + Calculated C 28 H 21 AuClN5NaS + : 714.095; found: 714.0757.

[0090] Example 2

[0091] This example prepared gold complex G2, 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, yield 70%). 1 H NMR (600 MHz, CDC13) δ 7.97 (d, J = 8.5 Hz, 1H), 7.85 (d, J = 6.8 Hz, 1H), 7.48 (t, J = 7.4 Hz, 1H).

[0095] 2. Synthesis of compound 2b:

[0096] Into a 250 mL reaction flask was added 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). Subsequently, 1,4-dioxane (100 mL) was added to the reaction flask. The mixture was refluxed at 105 °C for 24 hours. After the reaction was completed, the solvent was removed by evaporation under reduced pressure. The crude product was purified by column chromatography to give the desired yellow solid 2b (0.67 g, yield 51%). 1H NMR (600 MHz, CDC13) δ 8.18 (d, J = 6.2 Hz, 1H), 8.12 (d, J = 8.5 Hz, 1H), 7.63-7.56 (m, 1H), 1.45 (s, 12H).

[0097] 3. Synthesis of compound 2d:

[0098] Compound 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 refluxed at 80 °C for 24 h while monitoring the progress of the reaction 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 red solid 2d (0.26 g, yield 56%). Proton nuclear magnetic resonance (1H NMR) (600 MHz, CDC13) 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, CDC13) 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. Subsequently, DMF (50 mL) was added to the reaction flask and the mixture was heated to reflux for 24 h. After the reaction was completed, 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 h. The crude product was used directly for 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 iodomethane (1.42 g, 10.00 mmol) and DMF (50 mL) to the flask. The mixture was refluxed at 100 °C for 12 h. After completion of the reaction, 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 red solid 2f (2.32 g, yield 79%). 1 H NMR (600 MHz, CDC13) δ 9.00 (s, 1H), 7.91 (dd, J = 24.6, 7.2 Hz, 3H), 7.66 (d, J = 7.6 Hz, 2H), 7.59 (s, 1H), 7.57 (s, 1H), 7.47 (d, J = 8.2 Hz, 2H), 7.34 (t, J = 6.8 Hz, 2H), 7.30-7.18 (m, 6H), 7.18-7.12 (m, 1H), 4.19 (s, 3H). 13 C NMR (101 MHz, CDC13) δ 155.7, 153.4, 149.7, 146.7, 146.3, 135.6, 133.6, 133.0, 130.5, 130.0, 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 Ag20 (0.04 g, 0.15 mmol) were added to a 50 mL reaction flask, followed by DCM (30 mL) to the flask. The mixture was stirred at room temperature for 48 h in the dark. Then (tht)AuCl (0.10 g, 0.30 mmol) was added and stirring was continued for 12 h. The reaction mixture was filtered through celite and the filtrate was evaporated under reduced pressure. The crude product was purified by flash column chromatography on neutral alumina using DCM as eluent and then recrystallized from a DCM / n-hexane mixture to give the desired red solid G2 (0.14 g, yield 67%). 1H NMR (400 MHz, CDC13) δ 8.01 - 7.98 (m, 1H), 7.92 (d, J = 8.4 Hz, 2H), 7.71 - 7.68 (m, 2H), 7.61 (d, J = 8.8 Hz, 2H), 7.37 (t, J = 7.6 Hz, 2H), 7.32 - 7.25 (m, 8H), 7.16 (t, J = 7.4 Hz, 1H), 7.11 (s, 1H), 4.05 (s, 3H). 13 C NMR (101 MHz, CDC13) δ 170.8, 162.1, 155.7, 153.4, 148.4, 147.2, 146.7, 133.8, 132.7, 132.3, 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] This example prepared gold complex G3, the specific process is as follows:

[0107]

[0108] 1. Synthesis of compound 3b:

[0109] 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 bottle. Then a solvent system of THF (50 mL) and H2O (10 mL) was added to the bottle. 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 separated by extraction, and purified by silica gel column chromatography to obtain the target compound 3b as an orange solid (1.03 g, yield 40%).

[0110] 2. Synthesis of compound 3c:

[0111] Into a 100 mL reaction flask was added 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). To the flask was then added DMF (50 mL) and the mixture was heated to reflux for 24 hours. Upon completion of the reaction, 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 step without further purification.

[0112] 3. Synthesis of compound 3d:

[0113] Into a 100 mL reaction flask was added 3c (2.53 g, 5.00 mmol) followed by iodomethane (1.42 g, 10.00 mmol) and DMF (50 mL). The mixture was refluxed at 100 °C for 12 hours. Upon completion of the reaction, 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 yield red solid 3d (2.10 g, 65% yield). 1 H NMR (600 MHz, CDC13) δ 10.82 (s, 1H), 8.52 (s, 1H), 8.34 (s, 1H), 7.73 (d, J = 70.6 Hz, 4H), 7.06 (d, J = 89.8 Hz, 10H), 4.34 (s, 3H), 3.81 (s, 6H). 13 C NMR (151 MHz, CDC13) δ 156.56, 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] To a 50 mL reaction flask was added 3d (0.19 g, 0.30 mmol) and Ag20 (0.04 g, 0.15 mmol) followed by DCM (30 mL). The mixture was stirred at room temperature for 48 h in the dark. Then (tht)AuCl (0.10 g, 0.30 mmol) was added and stirring was continued for 12 h. The reaction mixture was filtered through celite and the filtrate was evaporated under reduced pressure. The crude product was purified by flash column chromatography on neutral alumina using DCM as eluent and then recrystallized from a DCM / n-hexane mixture to give the desired red solid G3 (0.12 g, 53% yield). 1 H NMR (600 MHz, CDC13) δ 8.20 (d, J = 7.6 Hz, 1H), 7.77 (dd, J = 32.2, 8.2 Hz, 3H), 7.61 (d, J = 1.9 Hz, 1H), 7.21 (d, J = 1.9 Hz, 1H), 7.16 (d, J = 8.9 Hz, 4H), 7.04 (d, J = 8.7 Hz, 2H), 6.88 (d, J = 8.9 Hz, 4H), 4.01 (s, 3H), 3.82 (s, 6H). 13 C NMR (151 MHz, CDC13) δ 172.22, 156.43, 154.12, 150.55, 149.67, 140.21, 135.66, 130.00, 127.90, 127.35, 126.75, 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] + Caled for C 30 H 25 AuClN5O2NaS + : 774.1161; found: 774.0986.

[0116] Example 4

[0117] This example prepared gold complex G4, the specific process is as follows:

[0118]

[0119] To a 50 mL reaction vial was added 1f (0.18 g, 0.30 mmol) and Ag20 (0.04 g, 0.15 mmol) followed by DCM (30 mL) to the vial. The mixture was stirred at room temperature for 48 h in the dark. NaCI (0.18 g, 0.30 mmol) and (tht)AuC6F5 (0.14 g, 0.30 mmol) were then added and stirred at room temperature overnight. The solution was filtered through celite and the filtrate was solvent removed under reduced pressure. The crude product was purified by flash column chromatography over neutral alumina using DCM as 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 (400 MHz, CDC13) δ 8.50 (d, J = 7.6 Hz, 1H), 7.90 - 7.78 (m, 4H), 7.31 (t, J = 7.6 Hz, 4H), 7.21 (dd, J = 8.6, 3.8 Hz, 7H), 7.09 (t, J = 7.2 Hz, 2H), 4.11 (s, 3H). 13 C NMR (101 MHz, CDC13) δ 173.1, 154.1, 150.3, 148.7, 147.3, 135.0, 130.2, 129.6, 129.5, 128.4, 126.3, 126.2, 125.2, 123.6, 123.0, 122.5, 121.9, 38.6. 19 F NMR (376 MHz, CDC13) 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] This example prepared gold complex G5, 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 into a 50 mL reaction bottle, then DCM (30 mL) was added into the bottle. The mixture was stirred at room temperature for 24 hours. The reaction mixture was filtered through celite, and the filtrate was removed solvent under reduced pressure. The crude product was purified by recrystallization with DCM / n-hexane mixture to give the desired red solid G5 (0.17 g, yield 76%). 1 H NMR (400 MHz, CD2CI2) δ 7.92 (d, J = 3.2 Hz, 1H), 7.91 (d, J = 3.2 Hz, 1H), 7.81 (d, J = 8.6 Hz, 2H), 7.62 (q, J = 3.8 Hz, 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 = 2.0 Hz, 1H), 7.27 (d, J = 8.0 Hz, 2H), 7.25-7.22 (m, 2H), 7.13-7.08 (m, 7H), 3.95 (s, 3H). 13 C NMR (101 MHz, CD2CI2) δ 155.8, 148.8, 147.0, 146.6, 134.2, 134.0, 132.4, 130.4, 130.4, 129.8, 129.7, 129.7, 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 (160 MHz, CD2CI2) δ 40.24 (s, PPh3), -144.52 (m, PF6 - ). HRMS: m / z: [M-PF6] + calcd for C 46 H 36 AuN5PS + : 918.2173; found: 918.2085.

[0124] Test Example 1

[0125] 1. Prepare the complex G1-G5 solution with a final concentration of 2 x 10 -5 mol·L -1 in DMF, and place it in a cuvette, and detect the UV-absorption spectrum thereof by using a Varian CARY 50 UV spectrometer. Figure 1UV-Vis absorption spectra of G1, G2 in THF / Hex mixed solution at 0% and 90% Hex content, respectively. Figure 1 UV-Vis absorption spectra of G3, G4 in THF / Hex mixed solution at 0% and 99% Hex content, respectively. Figure 1 UV-Vis absorption spectra of G5 in THF / Hex mixed solution at 0% and 70% Hex content, respectively. It can be seen that the complexes G1-G5 have obvious absorption peaks at 400-500 nm.

[0126] 2, Based on their absorption spectra, the fluorescence complexes G1-G5 are excited by 445 nm, 410 nm, 488 nm, 465 nm, 400 nm light, respectively. Figure 2 The fluorescence spectra of complexes G1-G5 (corresponding to G1, G2, G3, G4, G5, respectively) Figure 2 The fluorescence spectra of complexes G1-G5 (corresponding to G1, G2, G3, G4, G5, respectively) in THF / Hex mixed solution, with the addition of poor solvent Hex, the luminescence of the resulting system gradually increases, and reaches a maximum when the Hex content is 90vol%, 90vol%, 99vol%, 99vol%, 70vol%, respectively.

[0127] 3, DCFH-DA (2', 7'-Dichlorodihydrofluorescein diacetate) is used as a ROS active oxygen detection probe to detect the ROS generation of the synthesized fluorescence G1-G5 in solution (aggregated state). DCFH-DA (0.05 mL, 10 mM) is dissolved in 2 mL of 10 mM sodium hydroxide solution, continuously stirred (25°C, 30 minutes) for hydrolysis, and then neutralized with 10 mL of 0.01M phosphate buffer (PBS, pH 7.4), finally obtaining a 40 μM DCFH solution. Take the solution and mix with 10 μL of the sample to be tested (G1-G5, rose red RB or Ce6, all at a concentration of 10 μM) in a quartz cuvette. The sample is irradiated with white light (0.06 W / cm 2 ) light, and the fluorescence intensity is measured at 488 nm excitation wavelength and 525 nm emission wavelength using a fluorescence spectrophotometer (Horiba Fluoromax4). The results are shown in Figure 2a. Figure 3 As shown in Figure 2a, under white light irradiation, except for complexes G2 and G5, the remaining complexes can efficiently and rapidly generate ROS, which is significantly better than the commercial photosensitizer rose red (RB). The ROS generation efficiency of complex G3 is the highest, and the fluorescence intensity is much higher than that of other complexes and commercial dyes within the first 60s, indicating that it has the highest generation efficiency, followed by complex G1.

[0128] 4. Singlet oxygen detection (dimethylglyoxime anthracene ABDA method): ABDA (9,10-anthracene-diyl-bis(methylene) dimalonic acid) was used as a singlet oxygen detection probe. ABDA was dissolved in PBS to prepare a 10 mM stock solution, and 10 μL was mixed with complexes G1-G5, RB or Ce6 (all at a final concentration of 10 μM) in PBS (final volume 2 mL) and transferred to a quartz cuvette. After white light irradiation (0.06 W / cm2), the change in absorbance of ABDA at 378 nm was monitored by UV-visible spectrophotometry (Varian CARY 50), and the decay rate reflected the efficiency of singlet oxygen generation. The results are shown in Table 1. 2 Table 1. Singlet oxygen generation efficiency of complexes G1-G5, RB and Ce6. Figure 3 As shown in Table 1, under white light irradiation, complexes G3 and G5 can completely oxidize ABDA within the first 60 s, reducing its absorption to the baseline standard, while the commercial dyes RB and Ce6 require about 120 s to completely oxidize ABDA, indicating that G3 and G5 have significantly stronger singlet oxygen generation ability than RB and Ce6.

[0129] Example 6

[0130] This example uses complexes G1-G5 to image fungi (standard strains) in vitro, and the specific process is as follows:

[0131] Five clinically relevant Candida species (C. albicans, C. krusei, C. glabrata, C. tropicalis and C. parapsilosis) were selected. The above strains were provided by the Southern Hospital of Southern Medical University. The bacteria were cultured in YPD medium (37°C, 200 rpm) for 16 hours, centrifuged (4000 x g, 5 minutes) and resuspended in a solution containing G1-G5 (10 μM, 500 μL PBS), and incubated at 37°C for 30 minutes. 1 μL of stained bacteria solution was added to a glass slide, covered with a glass slide, and observed using a confocal microscope (Carl Zeiss, LSM 880 with airscan) (excitation wavelength 488 nm, emission wavelength 550-650 nm). As shown in Table 2, all complexes showed significant fluorescence imaging performance on common clinical fungi, including C. albicans, C. krusei, C. glabrata, C. tropicalis and C. parapsilosis. The present application can provide a new fluorescence detection method for clinical fungal detection. Figure 4

[0132] Example 7

[0133] This example uses complex G3 to study the co-localization effect with commercial dyes and the dye imaging performance, and the specific process 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 a, G1-G5 all showed certain antifungal activity (nearly 100% antibacterial efficiency could be achieved within 10 μΜ) for standard strains, among which G3 had the strongest antifungal activity (for Candida glabrata, Candida parapsilosis, Candida albicans and Candida tropicalis, 100% antibacterial efficiency could be achieved when the concentration of G3 was 7.5 μΜ). Meanwhile, compared with dark conditions, the antifungal effect of the complex was significantly enhanced after white light irradiation (100% antibacterial efficiency could be achieved when the concentration of G3 was 2.5 μΜ), indicating that the complex had strong photodynamic effect.

[0139] Example 9

[0140] This example uses complex G3 to destroy the structure of Candida albicans, and the specific process is as follows:

[0141] Scanning electron microscopy (SEM) and transmission electron microscopy (TEM) were used to observe the effect of G3 on the structure destruction of Candida albicans: Candida albicans was cultured in YPD medium for 24 hours, and then the concentration of the bacterial solution was adjusted to 2 x 10 5 CFU / mL and incubated with G3 (10 μΜ) for 30 minutes, followed by white light irradiation (0.06 W / cm 2 ) or dark treatment. Then, the fungi were collected by centrifugation at 4000 rpm for 10 minutes, fixed with 2.5% glutaraldehyde at 4°C overnight, and then fixed with 1% osmium tetroxide for 1-2 hours. The samples were dehydrated with different concentrations of ethanol (30%, 50%, 70%, 80%, 90%, 95% and 100%) for 15 minutes each. Then, the samples were treated with a mixture of ethanol and isoamyl alcohol (1:1) for 30 minutes, and then 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 ethanol gradient dehydration, the samples were treated with pure acetone for 20 minutes, followed by treatment with a mixture of embedding agent and acetone (1:1, v / v) for 1 hour, and then with a mixture of embedding agent and acetone (3:1, v / v) for 3 hours. Then, the samples were treated with pure embedding agent and heated at 70°C overnight. Finally, an ultramicrotome (LEICA EMUC7) was used to prepare ultrathin sections of 70-90 nm. After staining and drying, the sections were observed using a transmission electron microscope (TEM, HITACHI H-7650). As shown in a, b and c, the scanning electron microscopy images showed that the fungal surface collapsed and ruptured after G3 treatment, and the transmission electron microscopy images showed that the contents of the fungal cells were blurred and the cell wall and cell membrane were damaged. These results indicate that G3 can induce fungal death by destroying the internal and membrane structures, further confirming the fungicidal effect of the complex, which can provide a new treatment for clinical fungal infections. Figure 6 As shown in a, b and c, the scanning electron microscopy images showed that the fungal surface collapsed and ruptured after G3 treatment, and the transmission electron microscopy images showed that the contents of the fungal cells were blurred and the cell wall and cell membrane were damaged. These results indicate that G3 can induce fungal death by destroying the internal and membrane structures, further confirming the fungicidal effect of the complex, which can provide a new treatment 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. 2White light irradiation for 30 minutes, the untreated group as control. After treatment, the bacteria were collected by centrifugation (4,000 x g, 5 min, 4°C) and stored in liquid nitrogen. Lipidomics detection and analysis were completed by Shanghai Majorbio Biomedical Technology Co., Ltd. The samples were analyzed using Thermo UHPLC-Q ExactiveHF-XVanquish Horizon system, and the chromatographic column was Accucore C30 column (100 mm x 2.1 mm inner diameter, 2.6 μm; Thermo, USA). The raw data were analyzed by LipidSearch (Thermo, USA) software for lipid identification and quantification, and the subsequent analysis was completed on the Majorbio cloud platform (cloud.majorbio.com). The variable importance projection (VIP) value and t-test P value based on the OPLS-DA model were used to screen the differential metabolites, and the metabolites with VIP>1, P<0.05 and fold change>2 were identified as significant differential metabolites. The results are as follows Figure 9 As shown in a, b, OPLS-DA analysis showed that the lipids of C. albicans changed significantly before and after G3 treatment. More than 680 different lipids were analyzed, including 382 glycerophospholipids (GP), 146 glycerolipids (GL), 133 sphingolipids (SP), 15 fatty acids (FA) and 4 sterol lipids (ST) Figure 9 Further classification and statistical analysis based on the unsaturation of lipid chain showed that there were 433 saturated fatty acids (SFA), 458 monounsaturated fatty acids (MUFA), 415 polyunsaturated fatty acids (PUFA) and 218 odd fatty acyls (ODD), most of which were polyunsaturated fatty acids Figure 9 There were 36 lipids that changed significantly after G3 treatment of fungi, of which 12 lipids decreased and 24 lipids increased. Figure 9 The top 20 lipids with the largest content difference are shown in f, of which the most significant difference is phosphatidic acid (PA). As a precursor of phospholipids, PA is involved in the formation of biological membranes, and the decrease of PA can weaken the structural stability of the cell membrane. Among the significantly increased lipids, most of them are PUFAs, which are key substrates for lipid peroxidation and ferroptosis. These findings lay the foundation for G3 to induce fungal ferroptosis.

[0151] Example 12

[0152] This example studies the induction of fungal ferroptosis by complex G3 through TrxR activity inhibition, and the specific process is as follows:

[0153] C. albicans was cultured overnight to the logarithmic growth phase, and the concentration was adjusted to 1 x 10 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 Support of d). In addition, compound G3 was able to inhibit the activity of thioredoxin reductase (TrxR) in C. albicans in a dose-dependent manner (Fig. 2c). Figure 11 a). Given that TrxR plays a key role in the antioxidant defense system, its activity inhibition would disrupt the antioxidant defense mechanism of the cell, leading to an increase in the level of reactive oxygen species (ROS). Consistent with this, a significant accumulation of ROS was observed after G3 treatment (Fig. 2b). Figure 10 e, Figure 11 b). To further verify the regulatory role of TrxR in ferroptosis, cell viability and lipid peroxidation levels were evaluated after TrxR inhibition by TRi-1 (Fig. 2c-e). Figure 11 c-e). The experimental results showed that TRi-1 treatment led to a decrease in C. albicans survival rate and accumulation of lipid peroxidation products. These results confirmed that G3 could regulate the ferroptosis process of C. albicans by inhibiting the activity of TrxR.

[0154] Example 13

[0155] This example analyzes the GSEA analysis of complex G3 driving mitochondrial dysfunction, the specific process is

[0156] The sample pretreatment step is the same as the aforementioned SEM characterization method: the C. albicans cells treated with G3 (5 μM) combined with white light irradiation (0.06 W / cm 2 , 30 minutes) and untreated (control) were centrifuged (4,000 x g, 15 minutes), washed with PBS and then frozen in liquid nitrogen. RNA sequencing was completed by Shanghai Majorbio Biotech Co., Ltd. Sequencing was performed using the Illumina HiSeq4000 sequencing platform. Data were analyzed online through the Majorbio cloud platform (cloud.majorbio.com). High-quality data were obtained after quality control filtering, and differential expression genes (|fold change|>2, P<0.05) were screened and subjected to pathway enrichment analysis. The results are shown in Figure 12 a, b, GSEA analysis showed that G3 was located in mitochondria, which disrupted the pathways related to mitochondrial function, including the tricarboxylic acid cycle and oxidative phosphorylation.

[0157] Example 14

[0158] This example studies the effect of complex G3 on fungal mitochondrial dysfunction and oxidative stress, the specific process is:

[0159] C. albicans was cultured overnight to the logarithmic growth phase, and the concentration was adjusted to 5 x 10 6CFU / mL (MCF = 5). 500 μL of cell suspension was taken for four treatments: (1) PBS; (2) PBS + white light irradiation (0.06 W / cm 2 , 30 min); (3) G3 (10 μM); (4) G3 (10 μM) + white light irradiation (0.06 W / cm 2 , 30 min). After treatment, cells were collected by centrifugation at 4000 x g for 5 min and washed twice with PBS. JC-1 fluorescent probe (10 μg / mL) was used to stain cells at 37 °C in the dark for 30 min. After the final centrifugation, the stained cells were resuspended in PBS and immediately analyzed by confocal laser scanning microscopy (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 the red-to-green fluorescence density ratio relative to the control group. ATP content determination was performed using an enhanced ATP detection kit (Bi Yun Tian, China) according to the manufacturer's instructions. The mitochondrial ROS level in C. albicans cells treated with G3 (white light irradiation) was detected using the fluorescent probe MitoSOX. The C. albicans suspension was adjusted to 1 x 10 7 CFU / mL and divided into four groups for treatment: (1) PBS; (2) PBS + light irradiation (0.06 W / cm 2 , 30 min); (3) G3 (10 μM); (4) G3 (10 μM) + light irradiation (0.06 W / cm 2 , 30 min). After treatment, the bacterial bodies were collected by centrifugation at 4000 x g for 5 min and stained with MitoSOX (2 μM) for 30 min in the dark, followed by three washes with PBS. Imaging was performed using confocal microscopy (CLSM, LSM880, Carl Zeiss, Germany), MitoSOX: λ ex = 590 nm, λ em = 600-630 nm. The results are shown in Figs. 16c, d, which indicate that G3 significantly reduces mitochondrial membrane potential and ATP levels, indicating mitochondrial dysfunction. Figure 12 The results shown in Fig. 16e indicate that the ROS in the mitochondria increases, and the mitochondria undergo oxidative stress. Figure 12

[0160] Example 15

[0161] This example studies the complex G3 driving ferroptosis through mitochondrial dysfunction and oxidative stress in fungi. The specific process is as follows:

[0162] After treatment with mito-tempo, the lipid peroxidation level in C. albicans and the survival rate of the fungus were detected as described in Example 12 and Example 8. The results are shown in Fig. 17. Figure 12 ​As 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 / cm). 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) G3 (10μM) + white light irradiation (0.06W / cm²) for 30 minutes; (9) G3 (10μM); (10) G3 (10μM) + white light irradiation (0.06W / cm²) for 30 minutes; ... 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] BALB / c mice (body weight 18-22 g) were anesthetized by intraperitoneal injection of sodium pentobarbital (75 mg / kg body weight). The hair on the back of the mice was removed using a hair clipper, and after alcohol disinfection, a full-thickness skin defect was created on the back of the mice using a surgical knife. Each wound was inoculated with 10 μL of a Candida albicans suspension (1 x 10 8 CFU / mL), and after drying, the wound was covered with sterile, breathable adhesive tape. After 24 hours of 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 / cm 2 white light for 30 min); (3) voriconazole group (20 μM, 50 μL); (4) G3 group (20 μM, 50 μL); (5) G3 + light group (20 μM, 50 μL, 0.06 W / cm 2 white light for 30 min). The treatment method was direct application to the wound, which was repeated every 24 h, for a continuous treatment of 3 days. Body weight and wound area were monitored daily. On day 11, blood samples and wound tissue and organs were collected, and the mice were euthanized. The wound tissue was subjected to hematoxylin and eosin (H&E) staining, Masson staining, and immunohistochemical analysis of IL-10, IL-6, and CD31 (to assess neovascularization). Quantitative analysis was performed using ImageJ. H&E staining of the organs (heart, liver, spleen, lung, kidney) was performed to evaluate the biocompatibility of G3. The results are shown in Figure 14 a, b, which show that the voriconazole, G3, and G3 + light treatment groups significantly promoted the healing of the skin wounds in mice, and H&E and Masson staining showed that G3 + light treatment significantly reduced inflammatory cell infiltration, accelerated granulation tissue formation, and promoted collagen deposition (Figs. 3 Figure 14 c, d, h, i). In addition, G3 + light treatment significantly increased the expression of proinflammatory factors IL-6 and TNF-a (Figs. 3 Figure 14 e, f, j, k). At the same time, G3 + light treatment significantly promoted neovascularization, which was beneficial for the repair and healing of the wound (Fig. 3 Figure 14 g, l). These results indicate that G3 has the effect of controlling fungal infection of the wound and promoting wound healing, providing a new treatment for clinical fungal infection of the wound. In addition, H&E staining of the main organs (heart, liver, spleen, lung, kidney) after different treatments showed no obvious pathological damage, and there was no difference in morphology and structure (Fig. 3 Figure 15 a). The liver and kidney function indicators (ALT, AST, CREA, BUN) (Figs. 3 Figure 15 b-e) were within the normal range. During the treatment period, the body weight of the mice in each group also did not change significantly (Fig. 3 Figure 15In the middle f). These results show that G3 has good biocompatibility and safety, further supporting its safety in wound infection control. The present application can provide a new treatment regimen for clinically drug-resistant fungal skin wound infection.

[0169] Example 18

[0170] This example uses hemolysis experiment to study the in vivo biocompatibility of complex G3, the specific process is as follows:

[0171] Take fresh mouse blood, centrifuge at 1500 rpm for 15 minutes, discard the supernatant and wash the red blood cells with normal saline three times. Then dilute the red blood cells with normal saline 20 times to prepare the working suspension. Mix the red blood cell suspension with different concentrations (0-50 μM) of G3 compound with equal volume, and incubate in a 37°C constant temperature incubator for 24 hours. After incubation, centrifuge at 1500 rpm for 15 minutes to collect the supernatant. The experiment also sets up negative control (normal saline treatment group) and positive control (distilled water treatment group). Use the enzyme marker to measure the absorbance value of each group at 405 nm wavelength, and calculate the hemolysis rate according to the following formula: Hemolysis rate (%) = [(G3 treatment group absorbance value - negative control group absorbance value) / (positive control group absorbance value - negative control group absorbance value)] x 100%. The results are as follows Figure 15 As shown in the middle g, h, G3 treatment of red blood cells does not cause obvious hemolysis (hemolysis rate is less than 13% when the complex concentration is 50 μM), indicating that it has good in vivo biocompatibility.

[0172] Example 19

[0173] This example 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 the corneal epithelium was scratched. 10 μL of Candida albicans suspension (1 x 10 8 CFU / ml) was injected into the superficial stroma of the cornea to establish a model of fungal keratitis. 48 h later, according to the previous wound infection model, randomly divided into 5 treatment groups (n = 4): (1) PBS control group (50 μL); (2) PBS + light group (50 μL, 0.06 W / cm 2 white light irradiation for 30 min); (3) voriconazole group (20 μM, 50 μL); (4) G3 group (20 μM, 50 μL); (5) G3 + light group (20 μM, 50 μL, 0.06 W / cm 2White light irradiation for 30 minutes). The treatment method is to directly apply on the corneal surface, 3 times a day. Observe the cornea and take pictures every 2 days. The clinical severity is scored by 3 masked observers according to edema, surface regularity, neovascularization, opaque area and opaque density (each score 0-3, total score 0-15). On the 7th day, the corneal epithelial defect is evaluated with 2 μL of 0.5% sodium fluorescein. On the 10th day, the rats are euthanized, the corneas are washed with PBS, homogenized and spread on YPD plates at 37°C for 24h, and CFU counting is performed. At the same time, the corneal tissue is collected for histological analysis. In addition, the material is used to treat the corneas of healthy mice, and the biocompatibility is evaluated by H&E staining. After 3 days of continuous treatment, the control group corneas still show turbidity and edema, the G3 and voriconazole groups have moderate treatment effect, while the G3+light treatment group has significantly reduced corneal turbidity and edema Figure 16 a), the clinical score is the lowest Figure 16 d), fluorescein sodium staining shows that the corneal epithelial damage of the G3+light treatment group is the smallest (the lesion area is 0.32, while the lesion areas of the control group, light group, voriconazole, G3 group are 1, 0.96, 0.65, 0.45 respectively) Figure 16 b, e). The fungal count is the lowest, and the relative fungal load is as low as 0.12 (the fungal loads of the control group, light group, voriconazole, G3 group are 1, 0.99, 0.41, 0.25 respectively) Figure 16 c, f), the treatment effect is the best. H&E staining shows that the corneal structure of the control group is abnormal, and there are many inflammatory cells, while the corneal infiltration of the G3+light group is significantly reduced, the cornea is thin, and the swelling degree is reduced Figure 16 g). The results show that G3 has great potential and advantage in treating fungal keratitis. The present application can provide a new treatment scheme for clinically drug-resistant fungal corneal infection.

[0175] Example 20

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

[0177] 6-week-old female ICR mice (20-22g) were immunosuppressed by intraperitoneal injection of cyclophosphamide (200mg / kg) 3 days before infection. The mice were infected by intravenous injection of 100 μL of Candida albicans (5×10 6 CFU / ml). 24h later, according to the grouping of the skin wound infection model, the mice were randomly divided into 5 groups (n=4): (1) PBS control group (50 μL); (2) PBS+light group (50 μL, 0.06W / cm 2(3) voriconazole group (500 μM, 50 μL); (4) G3 group (500 μM, 50 μL); (5) G3+light group (500 μM, 50 μL, 0.06 W / cm 2 White light irradiation for 30 min). G3 (500 μM) was injected intravenously once every 24 h for 6 consecutive days. PBS and voriconazole (500 μM) were used as negative and positive controls, respectively. During the treatment, the mice were monitored for death, and the major organs (heart, liver, spleen, lung, kidney) of the dead mice were collected, homogenized in PBS, and plated on YPD agar for CFU counting. The surviving mice were euthanized after completion of the treatment regimen, and their organs were collected and fixed in 4% paraformaldehyde overnight, paraffin-embedded, and sectioned at 5 μm thickness for H&E staining. The results are shown in Figs. 6A-6F. Figure 17 In Fig. 6A, the survival rate of the control mice was low, and all of the mice died within 4 days after infection. G3 and voriconazole treatment significantly prolonged the survival time and survival rate of the infected mice. Meanwhile, G3 and G3+light reduced the fungal load in the organs of the mice (Figs. 6B-6F). Figure 17 In Fig. 6A, the survival rate of the control mice was low, and all of the mice died within 4 days after infection. G3 and voriconazole treatment significantly prolonged the survival time and survival rate of the infected mice. Meanwhile, G3 and G3+light reduced the fungal load in the organs of the mice (Figs. 6B-6F). Figure 17 H&E staining showed that G3 effectively reduced typical pathological changes, such as inflammatory infiltration and necrosis, especially in the kidney (Fig. 6G), indicating the good in vivo therapeutic effect of G3. The present application can provide a new treatment regimen for clinically drug-resistant fungal systemic infection.

[0178] The above examples are the preferred embodiments of the present application, but the embodiments of the present application are not limited by the above examples, and any changes, modifications, substitutions, combinations, simplifications made without departing from the spirit and principles of the present application should be equivalent replacement methods, and are included in the protection scope of the present application.

Claims

1. A compound of Formula I, Formula II, or a pharmaceutically acceptable salt thereof: wherein, R1, each R2, each R4are each independently selected from the group consisting of H, halogen, C1-C6alkyl, C1-C6alkoxy; said alkyl, alkoxy groups are optionally non-substituted, or substituted by one or more R a substituents; 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 of claim 1, wherein: each R3 is independently selected from F, Cl, Br, I, phenyl ring, biphenyl, diphenylphosphino, triphenylphosphino; wherein each phenyl is optionally non-substituted, or substituted with one or more R a substituted.

3. The compound of claim 1, wherein: the compound is selected from the following compounds:

4. A process for the preparation of said compound, characterized in that: comprising the following steps: a compound of Formula I is prepared by Suzuki coupling of a compound of Formula I-1 with a compound of Formula I-2, followed by reaction with imidazole, then quaternary amination with a compound of Formula I-3, and then exchange reaction; a compound of Formula II is prepared by Suzuki coupling of a compound of Formula II-1 with a compound of Formula II-2, followed by reaction with imidazole, then quaternary amination with a compound of Formula I-3, and then exchange reaction; wherein R1, R2, R4 are as defined in any one of claims 1-3; R5, R6 are independently selected from any one of boronic acid group, triflate group, triol borate group, borate ester group; X, X1, X2, X3, X4 are each independently selected from halogen.

5. A pharmaceutical composition comprising a compound of any one of claims 1-3, and optionally, a pharmaceutically acceptable excipient.

6. The pharmaceutical composition of claim 5, wherein: The pharmaceutical composition is a fungal fluorescence imaging agent.

7. Use of a compound of any one of claims 1-3 in the manufacture of a medicament for the prevention and / or treatment of fungal infection or disease, or a fluorescence probe.

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

9. Use according to claim 7, characterized in that: The fungal infection to be prevented and / or treated is selected from candidemia, invasive candidiasis, tinea capitis, tinea corporis, tinea pedis, onychomycosis, periungual mycosis, tinea versicolor, oral thrush, fungal keratitis, vaginal candidiasis, respiratory candidiasis, biliary candidiasis, esophageal candidiasis, urinary tract candidiasis, systemic candidiasis, mucocutaneous candidiasis, aspergillosis, mucormycosis, South American blastomycosis, North American blastomycosis, histoplasmosis, coccidioidomycosis, sporotrichosis, mycotic sinusitis, or chronic sinusitis.

10. Use according to claim 7, characterized in that: The medicament is used in a photodynamic therapy, in which the light irradiation conditions are: light intensity 0.01-2.0 W / cm 2 ; light irradiation time 10-60 min; preferably, the light source is white light of 300-700 nm.

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