N-heterocyclic carbene-gold (i) -alkynyl complex, and preparation method and application thereof

Abstract

The application discloses an NHC-gold (I)-alkynyl complex and a preparation method and application thereof. The structural general formula of the NHC-gold (I)-alkynyl complex is shown as formula I: wherein R1 in the formula I is selected from any one of hydrogen, a substituted or unsubstituted C1-C10 alkyl group,,, and R2 is selected from any one of hydrogen and a substituted or unsubstituted C1-C10 alkyl group; R3 is selected from any one of hydrogen, a halogen atom, a substituted or unsubstituted six-membered heterocycle, wherein the substituted group is a nitrogen atom or a methyl group; R4 and R5 are independently hydrogen, or R4 and R5 are connected to form an unsaturated six-membered ring. The NHC-gold (I)-alkynyl complex can generate active gold through an aggregation-induced ligand exchange reaction under the condition that no external catalyst is involved, the active gold has excellent catalytic activity and thiol reaction activity, and can be used for antitumor treatment.

Description

Technical Field

[0001] This invention relates to the field of compound technology, and in particular to an NHC-gold(I)-alkynyl complex, its preparation method and application. Background Technology

[0002] Gold is a common precious metal, and due to its excellent chemical and biological properties, related gold compounds are widely used in catalysis and disease treatment. Based on the strong relativistic effect of gold, the 6s orbital of the gold atom contracts and the 5d orbital expands, giving gold ions strong π-acidity. Therefore, gold ions can serve as mild and efficient catalysts for activating carbon-carbon unsaturated bonds. Utilizing the catalytic properties of gold, the luminescence of activated alkyne-based catalytic probes can be used to detect the formation of active gold species.

[0003] In related technologies, gold complexes, based on their ability to bind to biomolecules including enzymes and DNA, have been extensively studied as anticancer and antirheumatic drugs, and gold-based drugs have a significant impact on human health. Due to the thiophilic nature of gold, gold-based drugs such as auronoxine target TrxR (thioredoxin reductase), a highly expressed enzyme in tumor cells. However, in practical applications, it has been found that due to the presence of a large number of non-target thiol proteins under physiological conditions, including a large amount of albumin in the blood, these proteins can bind to gold, causing off-target effects. To address the problem of gold complexes being inactivated by non-target proteins before reaching the tumor target, many researchers have designed gold complexes as prodrugs, allowing them to exist stably in non-tumor tissues. Once inside tumor cells, the antitumor effect of the gold complexes is specifically activated by external stimuli (such as light, heat, ultrasound) or by palladium-catalyzed bio-orthogonal metal-transfer. However, these methods have certain limitations, such as limited light penetration and the fact that palladium can be inactivated by high concentrations of thiol species under physiological conditions. Therefore, a large amount of palladium catalyst is required to achieve the desired antitumor effect.

[0004] Based on this, the present invention aims to provide an N-heterocyclic carbene (hereinafter referred to as NHC)-gold(I)-alkynyl complex, its preparation method and application, which can produce active gold in tumor cells without the need for other catalysts. Summary of the Invention

[0005] This invention aims to at least solve one of the technical problems existing in the prior art. To this end, this invention proposes an NHC-gold(I)-alkynyl complex or its pharmaceutically acceptable salt, ester, solvate, or prodrug, which can catalyze the generation of active gold without the participation of an exogenous catalyst, and can be used to prepare antitumor drugs.

[0006] The present invention also proposes a pharmaceutical composition.

[0007] This invention also proposes a method for preparing NHC-gold(I)-acetylenic complexes.

[0008] The present invention also proposes the use of the above-described NHC-gold(I)-alkynyl complex as shown in Formula I, or its pharmaceutically acceptable salts, esters, solvates, and prodrugs, in the preparation of active gold.

[0009] The present invention also proposes the use of the above-described NHC-gold(I)-alkynyl complex as shown in Formula I, or its pharmaceutically acceptable salt, ester, solvate, or prodrug in the preparation of antitumor drugs.

[0010] In a first aspect, the present invention provides an NHC-gold(I)-alkynyl complex of formula I or a pharmaceutically acceptable salt, ester, solvate, or prodrug thereof. ; In formula I, R1 is selected from hydrogen, substituted or unsubstituted C1-C10 alkyl groups, , , Any one of them, wherein the substituted group is an azide group; The R2 is selected from hydrogen, substituted or unsubstituted C1 to C10 alkyl groups; The R3 is selected from any one of hydrogen, halogen atoms, substituted or unsubstituted six-membered heterocycles, wherein the substituted group is a nitrogen atom or a methyl group; R4 and R5 are either hydrogen atoms or are connected to form an unsaturated six-membered ring.

[0011] The NHC-gold(I)-acetylenic complex according to embodiments of the present invention has at least the following beneficial effects: The NHC-gold(I)-alkynyl complex of this invention can undergo aggregation-induced ligand exchange reaction. By adjusting the ratio of organic solvent and water / PBS in a mixed solvent, this invention demonstrates that the NHC-gold(I)-alkynyl complex exhibits solubility-dependent aggregation-induced ligand exchange properties. Specifically, under mixed solvent conditions with a low proportion of organic solvent, the complex does not dissolve well but can be well dispersed (i.e., it is within a certain solubility threshold). Under these conditions, intermolecular interactions occur in the complex molecules, leading to ligand exchange and the generation of active gold species. Furthermore, the active gold generated after the reaction of the NHC-gold(I)-alkynyl complex of this invention exhibits excellent catalytic and thiol-reactive activities, and can inhibit tumor cell proliferation, making it suitable for anti-tumor therapy.

[0012] In some embodiments of the present invention, R3 in Formula I is selected from any one of hydrogen, halogen atoms, or methyltetraazine groups.

[0013] In some embodiments of the present invention, the NHC-gold(I)-alkynyl complex is selected from any one of compounds of formula I-1 to I-9.

[0014] .

[0015] In some embodiments of the present invention, the NHC-gold(I)-alkynyl complex is selected from any one of compounds of formula I-5 to I-9.

[0016] In some embodiments of the present invention, the NHC-gold(I)-alkynyl complex is a compound of formula I-7 or formula I-8 (i.e., compound IMeTAGalAc-Au-PA or IMeTAGal-Au-PA).

[0017] The compounds IMeTAGalAc-Au-PA or IMeTAGalAc-Au-PA of the present invention can react with esterases and / or β-galactosidases expressed in tumor cells. Specifically, IMeTAGalAc-Au-PA can be progressively cleaved by esterases and β-galactosidases, and IMeTAGal-Au-PA can be cleaved by β-galactosidase. The solubility (hydrophilicity) of the products before and after cleavage decreases relative to the complex itself before cleavage, which helps to improve the aggregation effect, triggers ligand exchange reactions, and then generates active gold to inhibit tumor cell proliferation.

[0018] In some embodiments of the present invention, the pharmaceutically acceptable salt of the NHC-gold(I)-alkynyl complex includes any one of hydrochloride, nitrate, acetate, methanesulfonate, phosphate, citrate, fumarate, sulfate, succinate, tartrate, citrate, hydrobromide, hydroiodide, lactate, benzylsulfonate, cinnamate, salicylate, malonate, glutarate, and malate.

[0019] A second aspect of the present invention provides a method for preparing NHC-gold(I)-acetylene complexes as shown in Formulas I-7, comprising the following steps: In a solvent, the compound shown in Formula II is mixed with the compound shown in Formula III and reacted to obtain the product; wherein Formula I-7 is shown below: ; The structural formulas of the compound shown in Formula II and the compound shown in Formula III are as follows: .

[0020] The preparation method according to the embodiments of the present invention has at least the following beneficial effects: the preparation method of the present invention is simple, safe, and low in cost, and can be used for industrial production.

[0021] A third aspect of the invention provides a pharmaceutical composition comprising a therapeutically effective amount of the NHC-gold(I)-alkynyl complex of formula I as described in any one of the first aspects, or a pharmaceutically acceptable salt, ester, solvate, or prodrug thereof.

[0022] The pharmaceutical composition according to embodiments of the present invention has at least the following beneficial effects: the pharmaceutical composition of the present invention can inhibit the proliferation of tumor cells in vivo without palladium metal catalyst, and can be used as an anti-tumor drug.

[0023] In some embodiments of the present invention, the pharmaceutical composition further comprises compounds BCN-OOct and / or BCN-OAc, wherein the chemical structural formulas of BCN-OOct and BCN-OAc are as follows: .

[0024] The NHC-gold(I)-alkynyl complex of the present invention, when used in combination with compounds BCN-OOct and / or BCN-OAc, can significantly enhance the killing power against tumor cells and play a positive role in improving the efficacy of tumor treatment.

[0025] In some embodiments of the present invention, the pharmaceutical composition further includes pharmaceutically acceptable excipients.

[0026] In some embodiments of the present invention, the pharmaceutically acceptable excipients include at least one of solvents, excipients, diluents, binders, disintegrants, dispersants, flavoring agents, suspending agents, surfactants, isotonic agents, thickeners, emulsifiers, preservatives, flow aids, and lubricants.

[0027] In some embodiments of the present invention, the dosage form of the drug is any one of tablets, capsules, granules, injections, powder for injection, eye drops, liniments, suppositories, ointments, and aerosols.

[0028] In some embodiments of the invention, the excipient comprises water.

[0029] In some embodiments of the present invention, the filler includes at least one of starch and sucrose.

[0030] In some embodiments of the present invention, the adhesive includes at least one of cellulose derivatives, alginate, gelatin, and polyvinylpyrrolidone.

[0031] In some embodiments of the present invention, the wetting agent includes glycerin.

[0032] In some embodiments of the present invention, the disintegrant includes at least one of agar, calcium carbonate, and sodium bicarbonate.

[0033] In some embodiments of the present invention, the absorption enhancer includes a quaternary ammonium compound.

[0034] In some embodiments of the present invention, the surfactant includes hexadecyl alcohol.

[0035] In some embodiments of the present invention, the adsorbent carrier includes at least one of kaolin and soap clay.

[0036] In some embodiments of the present invention, the lubricant includes at least one of talc, calcium stearate, magnesium stearate, and polyethylene glycol.

[0037] In some embodiments of the present invention, the dosage form of the pharmaceutical composition is one of various dosage forms conventional in the art.

[0038] In some embodiments of the present invention, the dosage form of the pharmaceutical composition is a solid, semi-solid, or liquid form, which may be an aqueous solution, a non-aqueous solution, or a suspension.

[0039] In some embodiments of the present invention, the dosage form of the pharmaceutical composition is tablets, capsules, soft capsules, granules, pills, oral liquids, dry suspensions, drop pills, dry extracts, injections, or infusions.

[0040] It is understood that the pharmaceutical compositions of the present invention can be introduced into the body (such as muscle, intradermal, subcutaneous, venous, or mucosal tissue) by oral administration, injection, spraying, penetration, absorption, or physical or chemical mediated methods; or introduced into the body after being mixed or encapsulated with other substances.

[0041] A fourth aspect of the invention provides the use of the NHC-gold(I)-alkynyl complex of formula I as described in any of the first aspects, or a pharmaceutically acceptable salt, ester, solvate, or prodrug thereof, in the preparation of active gold.

[0042] According to the application of the embodiments of the present invention, at least the following beneficial effects are achieved: the NHC-gold(I)-alkynyl complex of the present invention can generate active gold through aggregation and ligand exchange reaction. The active gold has excellent catalytic activity and thiol reactivity and can be used for anti-tumor therapy. In some embodiments of the present invention, when the NHC-gold(I)-alkynyl complex is a compound of formula I-1 to I-6 or formula I-9, the method for preparing active gold includes: The NHC-gold(I)-alkynyl complex is reacted in a solvent and / or buffer solution to obtain the final product.

[0043] In some embodiments of the present invention, the solvent is a non-dissolving solvent, that is, a solvent that reduces the dispersion effect of the NHC-gold(I)-acetylenic complex.

[0044] Preferably, the solvent is selected from DMSO (dimethyl sulfoxide). d6 -At least one of DMSO (deuterated dimethyl sulfoxide) and MeCN (methyl ethyl carbonyl nitrogen).

[0045] In some embodiments of the present invention, the phosphate buffer is included.

[0046] In some embodiments of the present invention, the temperature of the reaction can be adjusted according to actual conditions, such as 25~40℃, preferably 37±2℃.

[0047] In some embodiments of the present invention, when the NHC-gold(I)-alkynyl complex is a compound represented by formula I-7 (IMeTAGalAc-Au-PA), the method for preparing active gold includes: The NHC-gold(I)-alkynyl complex is mixed with esterase and β-galactosidase in a solvent and / or buffer, and then purified after reaction.

[0048] In some embodiments of the present invention, when the NHC-gold(I)-alkynyl complex is a compound represented by formula I-8 (IMeTAGal-Au-PA), the method for preparing active gold includes: The NHC-gold(I)-alkynyl complex (IMeTAGal-Au-PA) is mixed with β-galactosidase in a solvent and / or buffer, and then purified after reaction.

[0049] A fifth aspect of the invention provides the use of the NHC-gold(I)-alkynyl complex as shown in Formula I as described in any of the first aspects, or a pharmaceutically acceptable salt, ester, solvate, or prodrug thereof, in the preparation of an antitumor drug.

[0050] According to the embodiments of the present invention, at least the following beneficial effects are observed: The NHC-gold(I)-alkynyl complex of the present invention can generate NHC-Au-NHC and an active gold product through ligand exchange, which has a strong killing effect on various tumor cells, including leukemia cells, and its inhibitory activity on tumor cells is further enhanced when it is used in combination with modified trans-cyclooctyne. Furthermore, cell experiments of the present invention have also revealed that the inhibitory activity of the compound IMeTAGalAc-Au-PA on cells is positively correlated with the expression level of GLB1 (the gene encoding β-galactosidase) in the cells; that is, the higher the expression level of β-galactosidase, the smaller the IC50 value of the compound IMeTAGalAc-Au-PA for that cell line, showing great promise for the treatment of cancers with high expression of β-galactosidase.

[0051] In some embodiments of the present invention, the tumor is a tumor capable of expressing β-galactosidase and / or esterase.

[0052] In some embodiments of the present invention, the tumor includes leukemia, lung cancer, colon cancer, and melanoma.

[0053] Furthermore, it is understood that the term "substituted or unsubstituted" in this invention refers to two situations in which one or more hydrogen atoms on the substituted group can be "substituted" or "not substituted" by one or more substituents. The number of substitutions ranges from monosubstituted to the maximum number of substitutions.

[0054] Other features and advantages of the present invention will be set forth in the following description. Attached Figure Description

[0055] The present invention will be further described below with reference to the accompanying drawings and embodiments, wherein: Figure 1 Here is the HRMS plot of compound 11 of the present invention; Figure 2 The HRMS image of the compound IMeTAGalAc-Au-PA of this invention is shown below. Figure 3 The image shows the HRMS plot of the compound IMeTAGal-Au-PA of this invention. Figure 4 The HRMS image of the compound IMeTAEtOH-Au-PA of this invention; Figure 5 The ligand exchange of the NHC-gold(I)-alkynyl complex 1a of this invention generates NHC-Au-NHC and PA-Au-PA. 1 H-NMR spectrum; Figure 6 This is a schematic diagram of the reaction mechanism by which the aggregation of the NHC-gold(I)-alkynyl complex induces ligand exchange to generate active gold in this invention; Figure 7 The figure shows the active gold-catalyzed Probe-1 luminescence generated by the solubility-related aggregation-induced ligand exchange of the NHC-gold(I)-alkynyl complex of the present invention. Figure 8 This is a schematic diagram of the stepwise enzymatic digestion process of the β-galactosidase-responsive gold (I) complex IMeTAGalAc-Au-PA of the present invention, and an HPLC characterization diagram of the extracellular enzymatic digestion process. Figure 9 The HPLC characterization of the degree of reaction of the β-galactosidase-responsive gold (I) complex IMeTAGalAc-Au-PA, the enzymatically digested products of IMeTAGal-Au-PA and IMeTAEtOH-Au-PA, with GSH under the same solvent conditions is shown. Figure 10 This is an HPLC characterization of the intracellular enzymatic digestion process of the β-galactosidase-responsive gold (I) complex IMeTAGalAc-Au-PA (100 μM) of the present invention. Figure 11 This is an HPLC characterization diagram of the intracellular enzymatic digestion process of the β-galactosidase-responsive gold (I) complex IMeTAGalAc-Au-PA (300 μM) of the present invention. Detailed Implementation

[0056] The following will describe the concept and technical effects of the present invention clearly and completely with reference to embodiments, so as to fully understand the purpose, features and effects of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, not all embodiments. Other embodiments obtained by those skilled in the art based on the embodiments of the present invention without creative effort are all within the scope of protection of the present invention.

[0057] The terms "preferred," "more preferably," etc., used in this invention refer to embodiments of the invention that provide certain beneficial effects under certain circumstances. However, other embodiments may also be preferred under the same or other circumstances. Furthermore, the description of one or more preferred embodiments does not imply that other embodiments are unavailable, nor is it intended to exclude other embodiments from the scope of this invention.

[0058] When a numerical range is disclosed herein, the range is considered continuous and includes the minimum and maximum values ​​of the range, as well as every value between the minimum and maximum values. Furthermore, when the range refers to integers, it includes every integer between the minimum and maximum values ​​of the range. Additionally, when multiple ranges are provided to describe a feature or characteristic, the ranges may be combined. In other words, unless otherwise specified, all ranges disclosed herein should be understood to include any and all subranges to which they are incorporated.

[0059] In the description of this invention, the reference term "and / or" includes all and any combination of one or more of the associated listed items.

[0060] In the description of this invention, the terms "one embodiment," "some embodiments," "illustrative embodiment," "example," "specific example," or "some examples," etc., refer to specific features, structures, materials, or characteristics described in connection with that embodiment or example, which are included in at least one embodiment or example of the invention. In this specification, the illustrative expressions of the above terms do not necessarily refer to the same embodiment or example. Furthermore, the specific features, structures, materials, or characteristics described may be combined in any suitable manner in one or more embodiments or examples.

[0061] In the embodiments of this invention, the RPMI 1640 medium and fetal bovine serum (FBS) used in cell experiments were both obtained from Gibco, USA, and penicillin and streptomycin were purchased from HyClone, USA. Unless otherwise specified, all chemicals were commercially available, primarily from Bidex Pharmaceuticals and Energie Chemicals.

[0062] The cell lines used in the experiment included PC9 (human lung cancer cells), HCC827 (human lung cancer cells), A549 (human lung cancer cells), HCT116 (human colon cancer cells), A375 (human melanoma cells), Caco-2 (human colorectal adenocarcinoma cells), THP-1 (human monocytic leukemia), K562 (human chronic myeloid leukemia), MOLM-13 (human acute myeloid leukemia), Jurkat (human T-lymphocytic leukemia), and C1498 (mouse acute myeloid leukemia). MOLM-13, C1498, and SKOV3 cells were obtained from Professor Zhang Xiaolei's research group at the School of Pharmaceutical Sciences, Sun Yat-sen University, while the remaining cells were obtained from the American Type Culture Collection (ATCC). In the embodiments or tests of this invention, the cancer cells are cultured in the following manner: Take the culture flask containing the cancer cells needed for the experiment, collect the cell suspension, centrifuge, discard the original culture medium, add 2 mL of fresh culture medium, and mix well by pipetting. Transfer 10 μL of the cell suspension to a cell counting plate and count the cells; plate the cells in a 96-well plate, seeding 10,000 cells per well and performing 3 replicates for each compound. Calculate the required cell suspension volume by counting the cells, dilute the required number of cells with culture medium to 10,000 cells per well, and seed the plate with 100 μL of culture medium. Mix well by pipetting, seed the cells using a pipette, and incubate in a CO2 incubator at 37 ℃. Label the cells.

[0063] In other embodiments where specific conditions are not specified, the procedures shall be performed under standard conditions or conditions recommended by the manufacturer. Reagents or instruments whose manufacturers are not specified are all commercially available products.

[0064] Example 1: Compound 1a This embodiment provides an NHC-gold(I)-alkynyl complex 1a (hereinafter referred to as compound 1a), the structural formula of which is shown in Formula I-1: .

[0065] The preparation method of the above compound 1a is as follows: ; The compounds 1,3-dimethylimidazole chloride (0.5 mmol, 66.3 mg), DMSAuCl (0.5 mmol, 147 mg), and K2CO3 (1.5 mmol, 207 mg) were added to a 25 mL Schlenk tube. A magnetic stir bar was added to the tube, and then 5 mL of acetone was added to dissolve the raw materials. The reaction tube was transferred to an oil bath and heated to 60 °C. The reaction was carried out for 1 h. After the reaction was completed, the mixture was filtered, and the filtrate was evaporated to dryness to obtain the crude product IDM-Au-Cl.

[0066] The characterization data of IDM-Au-Cl are as follows: 1 H NMR (400 MHz, Chloroform- d ) δ 6.93 (s, 2H), 3.83 (s, 6H).

[0067] IDM-Au-Cl (0.2 mmol, 66 mg), phenylacetylene (1.2 eq., 0.24 mmol), and NaOMe (5 eq. 1 mmol) were dissolved in 15 mL MeOH and reacted at 80 °C for 1 h. The solid obtained by rotary evaporation was redissolved in IDM and filtered. The solution was concentrated to ~1 mL, and recrystallized from n-pentane to give a white solid compound 1a.

[0068] The characterization data of compound 1a are as follows: 1 H NMR (400 MHz, DMSO- d 6) δ 7.39 (d, J = 2.3 Hz, 2H), 7.25 (d, J = 4.0Hz, 3H), 7.19 (m, 2H), 3.76 (s, 6H).

[0069] Example 2: Compound FPA This embodiment provides an NHC-gold(I)-alkynyl complex FPA (hereinafter referred to as compound FPA), whose structural formula is shown in Formula I-2: .

[0070] The preparation method of the above compound FPA is as follows: ; IDM-Au-Cl (0.2 mmol, 66 mg), p-fluorophenylacetylene (1.2 eq., 0.24 mmol), and NaOMe (5 eq. 1 mmol) were dissolved in 15 mL MeOH and reacted at 80 °C for 1 h. The solid obtained by rotary evaporation was redissolved in IDM and filtered. The solution was concentrated to ~1 mL, and recrystallized from n-pentane to give a yellowish-white solid compound FPA.

[0071] The characterization data of compound FPA are as follows: 1 H NMR (400 MHz, DMSO- d 6) δ 7.39 (s, 2H), 7.32 – 7.26 (m, 2H), 7.08 (dd, J = 10.0, 7.9 Hz, 2H), 3.75 (s, 6H).

[0072] Example 3: Compound Nap This embodiment provides an NHC-gold(I)-alkynyl complex Nap (hereinafter referred to as compound Nap), the structural formula of which is shown in Formula I-3: .

[0073] The preparation method of the above compound Nap is as follows: ; IDM-Au-Cl (0.2 mmol, 66 mg), 1-ethynylnaphthalene (1.2 eq., 0.24 mmol), and NaOMe (5 eq. 1 mmol) were dissolved in 15 mL MeOH and reacted at 80 °C for 1 h. The solid obtained by rotary evaporation was redissolved in IDM-Au-Cl, filtered, and the solution was concentrated to ~1 mL. Recrystallization was performed with n-pentane to give a white solid compound Nap.

[0074] The characterization data of compound Nap are as follows: 1 H NMR (400 MHz, Chloroform- d ) δ 8.66 (d, J = 8.3 Hz, 1H), 7.78 (d, J = 7.9 Hz, 1H), 7.70 (dd, J = 13.1, 7.8 Hz, 2H), 7.51 (t, J = 7.4 Hz, 1H), 7.46 (d, J = 7.5 Hz, 1H), 7.36 (t, J = 7.7 Hz, 1H), 6.88 (s, 2H), 3.87 (s, 6H).

[0075] Example 4: Compound Oct This embodiment provides an NHC-gold(I)-alkynyl complex Oct (hereinafter referred to as compound Oct), whose structural formula is shown in Formula I-4: .

[0076] The preparation method of the above compound Oct is as follows: ; Compound 1-methyl-3-octylimidazolium chloride (0.5 mmol, 115.4 mg), DMSAuCl (0.5 mmol, 147 mg), and K₂CO₃ (1.5 mmol, 207 mg) were added to a 25 mL Schlenk tube with a magnetic stir bar. 5 mL of acetone was added to dissolve the starting material. The reaction tube was transferred to an oil bath and heated to 60 °C for 1 h. After the reaction was complete, the mixture was filtered, and the filtrate was evaporated to dryness to obtain the crude product Oct-Au-Cl, which was an oily compound. All of the crude product was added to the reaction tube, along with phenylacetylene (0.5 mmol, 51 mg) and NaOMe (5 eq. 2.5 mmol) dissolved in 15 mL of MeOH. The reaction was carried out at 80 °C for 1 h. The solid obtained was evaporated to dryness, redissolved in DCM, filtered, and the solution was concentrated to ~1 mL. Recrystallization from n-pentane yielded a white solid.

[0077] The characterization data of compound Oct are as follows: 1 H NMR (400 MHz, Chloroform- d ) δ 7.51 (dt, J = 8.2, 1.2 Hz, 2H), 7.25– 7.19 (m, 2H), 7.19 – 7.15 (m, 1H), 6.92 – 6.87 (m, 2H), 4.18 (t, J = 7.3Hz, 2H), 3.85 (s, 3H), 1.83 (d, J = 7.7 Hz, 2H), 1.35 – 1.24 (m, 10H), 0.92 –0.84 (m, 3H).

[0078] Example 5: Compound N3 This embodiment provides an NHC-gold(I)-alkynyl complex N3 (hereinafter referred to as compound N3), the structural formula of which is shown in Formula I-5: .

[0079] The specific reaction process is as follows: .

[0080] The specific preparation process is as follows: The synthetic methods of compounds 1 and 2 are reported in the references. Specifically, compound 1 was synthesized by the method described in the reference "SUN J, WANG J, CHENG W, et al. Chitosan functionalized ionic liquid as a recyclable biopolymer-supported catalyst for cycloaddition of CO2 [J]. Green Chemistry, 2012, 14(3): 654-60."; compound 2 was synthesized by the method described in the reference "SAK H, MAWICK M, KRAUSE N. Sustainable Gold Catalysis in Water Using Cyclodextrin-tagged NHC-Gold Complexes [J]. ChemCatChem, 2019, 11(23): 5821-9.".

[0081] Compound 2 (0.5 mmol, 116 mg), DMSAuCl (0.5 mmol, 147 mg), and K2CO3 (1.5 mmol, 207 mg) were added to a 25 mL Schlenk tube with a magnetic stir bar. Then, 5 mL of acetone was added to dissolve the starting material. The reaction tube was transferred to an oil bath and heated to 60 °C for 1 h. After the reaction was complete, the mixture was filtered, and the filtrate was evaporated to dryness to obtain crude product 3. All of crude product 3 was added to a 25 mL Schlenk tube, along with phenylacetylene (0.5 mmol, 51 mg) and sodium methoxide (1 mmol, 51 mg), followed by 5 mL of methanol. The reaction tube was transferred to an oil bath and heated to 80 °C for 1 h. After the reaction was complete, the mixture was evaporated to dryness, redissolved, and filtered. The filtrate was recrystallized in a MeOH / diethyl ether system to obtain product N3 as a white powder with a yield of 45%.

[0082] The characterization data of product N3 are as follows: 1 H NMR (400 MHz, DMSO- d6 ) δ 7.61 (d, J = 1.9 Hz, 1H), 7.55 (d, J =1.9 Hz, 1H), 7.21 – 7.19 (m, 3H), 7.15 – 7.08 (m, 2H), 4.44 – 4.40 (m, 2H), 3.88 (s, 3H), 3.84 (t, J = 5.6 Hz, 2H). 13 C NMR (126 MHz, DMSO- d6 ) δ 183.75,167.52, 137.66, 131.51, 128.53, 125.70, 124.10, 122.64, 102.59, 51.42, 50.08,38.04.

[0083] Example 6: Compound Tz This embodiment provides an NHC-gold(I)-alkynyl complex Tz (hereinafter referred to as compound Tz), the structural formula of which is shown in Formula I-6: .

[0084] The specific reaction process is as follows: .

[0085] The specific preparation process is as follows: Compound 4 was synthesized according to the method reported in the literature “XIONG H, GU Y, ZHANG S, et al. Iridium-catalyzed C–Hamidation of s-tetrazines [J]. Chemical Communications, 2020, 56(34): 4692-5.”

[0086] The pink powder of compound 4 (0.15 mmol, 29 mg), IDM-Au-Cl (0.15 mmol, 49 mg), and sodium methoxide (0.3 mmol, 16.2 mg) were dissolved in 5 mL of methanol. The reaction tube was incubated at 80 °C for 1 h. After the reaction was completed, the solution was evaporated to dryness, redissolved, filtered, concentrated, dissolved in a small amount of DCM, and recrystallized with n-pentane to give compound Tz as a rose-red powder with a yield of 73%.

[0087] The characterization data of compound Tz are as follows: 1 H NMR (400 MHz, DMSO-d6) δ 8.36 (d, J = 8.4 Hz, 2H), 7.52 (d, J =8.4 Hz, 2H), 7.41 (s, 2H), 3.78 (s, 6H), 2.98 (s, 3H). 13 C NMR (126 MHz, Chloroform-d) δ 187.65, 166.74, 164.07, 133.67, 133.09, 130.43, 129.22,127.46, 121.80, 104.89, 37.93, 21.13.

[0088] Example 7: Compound IMeTAGalAc-Au-PA This embodiment provides an NHC-gold(I)-alkynyl complex IMeTAGalAc-Au-PA (hereinafter referred to as compound IMeTAGalAc-Au-PA), the structural formula of which is shown in Formula I-7: .

[0089] The specific reaction process is as follows: ; The specific preparation process is as follows: (1) Compounds 5, 6, and 7 were synthesized according to the methods reported in the literature. Specifically: Compound 5 was prepared according to the reference "BAUMANN K, KORDIĆ L, MOČIBOB M, et al. Synthesis and In Vitro Screening of Novel Heterocyclic β-d-Gluco- and β-d-Galactoconjugates as Butyrylcholinesterase Inhibitors [J / OL] 2019, 24(15)"; Compound 6 was prepared according to the reference "BABIUCH K, DAG A, ZHAO J, et al. Carbohydrate-Specific Uptake of Fucosylated Polymeric Micelles by Different Cancer CellLines [J]. Biomacromolecules, 2015, 16(7): 1948-57"; Compound 7 was prepared according to the reference "HOOGBOOM J, SWAGER T M. Increased Alignment of Electronic Polymers in Liquid Crystals via Hydrogen Bonding Extension [J]. Journal of the American Chemical Society, 2006, 128(47): 15058-9".

[0090] (2) Compound 7 (5.5 mmol, 1.51 g) and N-methylimidazole (5 mmol, 0.41 g) were dissolved in 10 mL of acetonitrile and the mixture was heated in an oil bath under reflux for 24 h. After the reaction was completed, the solvent was evaporated and the crude product was recrystallized in MeOH / diethyl ether to obtain compound 8 as brownish-yellow crystals with a yield of 46%.

[0091] The characterization data for compound 8 are as follows: 1 H NMR (400 MHz, DMSO-d6) δ 9.17 (s, 1H), 7.77 (t, J = 1.8 Hz, 1H), 7.75 (t, J = 1.8 Hz, 1H), 5.31 (s, 2H), 3.89 (s, 3H), 1.04 (d, J = 3.0 Hz,21H). 13C NMR (126 MHz, Chloroform-d) δ 137.04, 124.02, 121.05, 96.22, 91.97, 40.96, 37.18, 18.46, 10.89.

[0092] (3) Compound 8 (0.25 mmol, 89 mg), DMSAuCl (0.25 mmol, 74 mg), and K2CO3 (0.5 mmol, 69 mg) were added to a 25 mL Schlenk tube. A magnetic stir bar was added to the tube, and then 5 mL of acetone was added to dissolve the raw materials. The reaction tube was transferred to an oil bath and heated to 60 °C. The reaction was carried out for 3 h. After the reaction was completed, the mixture was filtered, the filtrate was evaporated to dryness, and a small amount of DCM was added to redissolve the filtrate. The mixture was recrystallized from n-pentane to obtain compound 9 as a white powder with a yield of 68%.

[0093] The characterization data for compound 9 are as follows: 1H NMR (400 MHz, Chloroform-d) δ 7.25 (d, 1H), 6.97 (d, J = 2.0 Hz,1H), 5.07 (s, 2H), 3.84 (s, 3H), 1.07 (m, J = 1.9 Hz, 21H). 13C NMR (126 MHz, Chloroform-d) δ 183.93, 123.30, 122.17, 120.78, 119.73, 98.89, 98.28, 89.88, 41.83, 41.71, 38.71, 38.33, 18.54, 11.03.

[0094] (4) Compound 9 (0.17 mmol, 87 mg) was added to a 25 mL Schlenk tube, followed by phenylacetylene (0.187 mmol, 19 mg) and sodium methoxide (0.34 mmol, 19 mg). Then, 5 mL of methanol was added. The reaction tube was transferred to an oil bath and heated to 80 °C. The reaction was allowed to proceed for 1 h. After the reaction was completed, the solution was evaporated to dryness, redissolved, and filtered. The filtrate was evaporated to dryness to obtain the crude product of compound 10. Compound 10 was completely dissolved in DMF, and an appropriate amount of CsF was added. The reaction was allowed to proceed at room temperature for 2 h. After the reaction was completed, the solution was evaporated to dryness, redissolved in DCM, and filtered. The filtrate was collected, concentrated, and recrystallized from DCM / n-pentane to obtain compound 11 as a yellowish-white powder with a yield of 56%.

[0095] The HRMS graph of compound 11 is shown below. Figure 1 As shown, its characterization data is as follows: 1H NMR (400 MHz, Chloroform-d) δ 7.69 (t, J = 6.5 Hz, 1H), 7.51 (d, J= 7.5 Hz, 2H), 7.22 (d, J = 7.4 Hz, 1H), 7.18 (d, J = 7.2 Hz, 1H), 7.12 (d, J= 1.9 Hz, 1H), 6.94 (d, J = 1.9 Hz, 1H), 5.62 (d, J = 6.4 Hz, 2H), 3.88 (s,3H), 3.87 – 3.83 (m, 1H). (5) Compound 11 (0.1 mmol, 41.8 mg), compound 6 (0.101 mmol, 42.15 mg), CuI (0.01 mmol, 1.9 mg), and TEA (0.04 mmol, 5.5 μL) were added to a 25 mL Schlenk tube under a nitrogen atmosphere. 5 mL of acetonitrile was added to dissolve the raw materials. After reacting at room temperature for 24 h, the solvent was evaporated and the solution was passed through an Al2O3 column (DCM / MeOH=150:1) to obtain compound IMeTAGalAc-Au-PA as a white powder with a yield of 20%.

[0096] HRMS graph of compound IMeTAGalAc-Au-PA is shown below Figure 2 As shown, its characterization data is as follows: 1H NMR (400 MHz, Chloroform-d) δ 8.35 – 8.32 (m, 1H), 7.64 (t, J =6.5 Hz, 1H), 7.52 – 7.39 (m, 1H), 7.34 (t, J = 7.7 Hz, 2H), 7.25 – 7.20 (m,1H), 7.18 (d, J = 2.0 Hz, 1H), 7.01 (d, J = 2.0 Hz, 1H), 5.65 (d, J = 6.5 Hz,2H), 5.37 (dd, J = 3.5, 1.2 Hz, 1H), 5.17 (dd, J = 10.5, 8.0 Hz, 1H), 4.95(dd, J = 10.5, 3.5 Hz, 1H), 4.80 (ddd, J = 14.2, 8.0, 6.7 Hz, 1H), 4.66 (ddd,J = 13.6, 6.4, 4.5 Hz, 1H), 4.51 (d, J = 8.0 Hz, 1H), 4.41 (ddd, J = 10.9,6.7, 4.4 Hz, 1H), 4.26 (ddd, J = 10.4, 7.9, 6.4 Hz, 1H), 4.13 (dd, J = 6.7,2.5 Hz, 2H), 3.94 (s, 3H), 3.91 – 3.85 (m, 1H), 2.12 (s, 3H), 2.03 (s, 3H), 1.94 (s, 3H), 1.88 (s, 3H). HRMS (Exact mass 835.21, found 836.2191 (+1)).

[0097] Example 8: Compound IMeTAGal-Au-PA This embodiment provides an NHC-gold(I)-alkynyl complex IMeTAGal-Au-PA (hereinafter referred to as compound IMeTAGal-Au-PA), the structural formula of which is shown in Formula I-8: .

[0098] The reaction process is as follows: .

[0099] Specific preparation methods include: Compound IMeTAGalAc-Au-PA (0.05 mmol, 41.78 mg) and sodium methoxide (0.1 mmol, 5.4 mg) were added to a 25 mL reaction flask. The starting material was dissolved in 5 mL of methanol and reacted at room temperature for 3 h. The pH was then neutralized to neutral by adding 1 M hydrochloric acid. The solvent was evaporated and the solution was passed through a silica gel column (DCM / MeOH = 5:1) to obtain compound IMeTAGal-Au-PA as a white powder with a yield of 90%.

[0100] The HRMS graph of compound IMeTAGal-Au-PA is shown below. Figure 3 As shown, its characterization data is as follows: HRMS (Exact mass 667.17, found 668.1773 (+1)).

[0101] Example 9: Compound IMeTAEtOH-Au-PA This embodiment provides an NHC-gold(I)-alkynyl complex IMeTAEtOH-Au-PA (hereinafter referred to as compound IMeTAEtOH-Au-PA), the structural formula of which is shown in Formula I-9: .

[0102] The reaction process is as follows: .

[0103] Specific preparation methods include: Azide ethanol was prepared according to the reference "KNIGHT NML, THOMPSON JDF, PARKINSON JA, et al. Iridium-Catalysed C(sp3)−H Activation and Hydrogen Isotope Exchange via Nitrogen-Based Carbonyl Directing Groups [J]. Advanced Synthesis & Catalysis, 2024, 366(11): 2577-86." Azide ethanol (0.2 mmol, 17.4 mg), compound 11 (0.2 mmol, 83.65 mg), CuI (0.02 mmol, 3.8 mg), and TEA (0.08 mmol, 11 μL) were added to a 25 mL Schlenk tube under a nitrogen atmosphere. 5 mL of acetonitrile was added to dissolve the starting material. After reacting at room temperature for 24 h, the solvent was evaporated, and the solution was passed through an Al₂O₃ column (DCM / MeOH = 150:1) to obtain compound IMeTAGalAc-Au-PA as a yellowish-white powder, with a yield of 68%.

[0104] The HRMS graph of compound IMeTAEtOH-Au-PA is shown below. Figure 4 As shown, its characterization data is as follows: 1 H NMR (400 MHz, Chloroform-d) δ 8.32 (d, J = 7.6 Hz, 1H), 7.60 (t, J= 6.5 Hz, 1H), 7.35 (t, J = 7.6 Hz, 2H), 7.25 – 7.19 (m, 2H), 7.16 (d, J =1.9 HRMS (Exact mass) 505.12, found 506.1251 (+1)).

[0105] Example 1: Verification of ligand exchange reaction of NHC-gold(I)-alkynyl complex 1a This test example uses the NHC-gold(I)-alkynyl complex 1a from Example 1 to verify whether the NHC-gold(I)-alkynyl complex can undergo ligand exchange, and the products generated by ligand exchange. The specific process is as follows: Weigh 2.5 nmol of compound 1a and dissolve it in 200 μL of DMSO. Add another 200 μL of PBS and shake at 37 °C for 12 h. After the reaction, add 400 μL of DMSO to dissolve the insoluble matter in the EP tubes, and then add another 200 μL of d6-DMSO. Take 250 μL of liquid from each EP tube and dilute it with 300 μL of DMSO. Detect the difference in the proton NMR spectra before and after the reaction using 1H-NMR.

[0106] Test results as follows Figure 5As shown, the 1H-NMR spectrum reveals a significant difference in the proton NMR spectra of compound 1a before and after the reaction. The solvent conditions used in the reaction ensured that compound 1a was in a poorly soluble state. Under these conditions, it can be inferred that intermolecular interactions occur in compound 1a, leading to ligand exchange and the formation of the ligand exchange products NHC-Au-NHC and PA-Au-PA, with the following structural formulas: .

[0107] Example 2: Verification of the catalytic effect of the active gold product This study investigates whether the active gold formed after ligand exchange in the above-mentioned NHC-gold(I)-alkynyl complexes (such as compound 1a, compound FPA, compound Nap, and compound Oct) can possess catalytic activity, such as catalyzing the luminescence of the coumarin precursor probe Probe-1 (a gold-catalyzed luminescent probe that can be used to detect active gold species). The specific experimental procedure is as follows: Seven EP tubes were taken from each group, and 1 mL of MeCN / PBS mixed solvent was added to each tube (the proportion of MeCN in the solvent was 100%, 80%, 60%, 40%, 20%, 10%, and 0%). Then, 20 μM Au and 100 μM Probe-1 were added, and the EP tubes were placed at 37 °C and shaken for 24 h. The reaction mechanism is illustrated in the diagram below. Figure 6 As shown.

[0108] Test results as follows Figure 7 As shown, NHC-gold(I)-alkynyl complexes 1a, FPA, Nap, and Oct can all catalyze Probe-1 luminescence in mixed solvents with low MeCN content.

[0109] Furthermore, based on the above detection example 1, it can be seen that ligand exchange of compound 1a will generate two compounds, NHC-Au-NHC and PA-Au-PA. Further research found that NHC-Au-NHC cannot catalyze the luminescence of Probe-1, while PA-Au-PA can. This indicates that ligand exchange of compound 1a can produce the active gold species PA-Au-PA. Combined with the above experiments, it can be seen that compounds FPA, Nap, and Oct can all achieve comparable technical effects.

[0110] Example 3: Detection of the inhibitory effect of compounds N3 or Tz on tumor cells This test case investigated the inhibitory effects of compound N3 or compound Tz in combination with modified trans-cyclooctylene on various tumor cells. The specific steps are as follows: Tumor cells were seeded at 5000 cells per well in 96-well plates. After 24 h, the concentration of trans-cyclooctyne was fixed at 25 μM, and culture medium containing different concentrations of compounds N3 or Tz was added. After incubation at 37 ℃ in a CO2 incubator for 48 h, 15 μL of LTT solution was added to each well, and the plates were incubated for another 3 h. The 96-well plates were then removed, and 100 μL of DMSO was added to each well to dissolve the formazan. The absorbance at 490 nm was measured using a microplate reader, and the IC50 value of the compounds was calculated.

[0111] Table 1: Toxicity of compounds Tz / N3 combined with BCN on different cell lines

[0112] Note: The cell viability of BCN-OOct or BCN-OAc at 25 μM is >90%. The chemical structural formulas of BCN-OOct and BCN-OAc are as follows: .

[0113] The test results showed that compounds N3 or Tz had a certain inhibitory effect on tumors, while the combination of BCN-OOct or BCN-OAc further enhanced the toxicity to various tumor cells. It is speculated that this is because the cyclooctyne moiety introduced after the click chemical reaction is more hydrophobic, so the gold complex after the reaction is more likely to undergo ligand exchange to produce active gold.

[0114] Example 4: Detection of compound IMeTAGalAc-Au-PA via enzymatic digestion reaction This assay tested whether the compound IMeTAGalAc-Au-PA could be stepwise cleaved by esterase and β-galactosidase. The specific steps are as follows: 200 μM of compound IMeTAGalAc-Au-PA was added to 1 mL of PBS. Esterase 10 U / mL was added and the reaction was carried out at 37 °C for 3 h. Then β-Gal 5 U / mL was added and the reaction was continued at 37 °C overnight. After the reaction was stopped, the protein was precipitated with acetone and the supernatant was analyzed by HPLC.

[0115] Test results as follows Figure 8 As shown, HPLC results indicate that compound IMeTAGalAc-Au-PA can be first cleaved by Esterase to generate compound IMeTAGal-Au-PA, and compound IMeTAGal-Au-PA can be further cleaved by β-galactosidase to generate IMeTAEtOH-Au-PA.

[0116] Example 5: Detection of the activity of the active gold thiol group in the enzyme digestion product. This assay example tested the products of IMeTAGalAc-Au-PA after enzymatic digestion, namely, IMeTAGal-Au-PA and IMeTAEtOH-Au-PA, and compared their reactivity with glutathione (GSH) under the same solvent conditions. The specific steps are as follows: 200 μM of compound IMeTAGal-Au-PA or compound IMeTAEtOH-Au-PA was added to 500 μL of a DMF / PBS mixed solvent. The DMF content in the solvent was 10%, 2%, or 90% (including the volume of Au stock solution added), and it contained 10 mM GSH (equivalent to triethylamine to neutralize pH). The reaction was carried out at 37 °C for 24 h, and then 500 μL of DMF was added to dissolve the residue. The HPLC analysis was performed.

[0117] HPLC detection results are as follows Figure 9 As shown, under the same solvent conditions, the remaining proportion of compound IMeTAEtOH-Au-PA is less than that of compound IMeTAGal-Au-PA, meaning that compound IMeTAEtOH-Au-PA reacts with GSH at a greater rate than compound IMeTAGal-Au-PA. This indicates that the hydrophilicity of the product IMeTAEtOH-Au-PA after enzymatic digestion is reduced compared to that of IMeTAGal-Au-PA before digestion, making it more prone to ligand exchange to generate the active gold PA-Au-PA. The generated active gold exhibits thiol reactivity, demonstrating that compound IMeTAEtOH-Au-PA reacts with GSH to a greater extent than compound IMeTAGal-Au-PA.

[0118] Example 6: Cell Experiment This assay detected the levels of compound IMeTAGalAc-Au-PA and its enzyme digestion products in K562 cells with increasing incubation time. The specific steps are as follows: K562 cells were seeded in 10 cm cell culture dishes and incubated for 2 h, 4 h, 8 h, and 12 h after administration of 100 μM of compound IMeTAGalAc-Au-PA, or for 1 h, 2 h, 4 h, and 8 h after administration of 300 μM. Cells were then collected by centrifugation, washed three times with PBS, and lysed by repeated freeze-thaw cycles at -80 ℃ three times. 600 μL of ice acetone was added to each EP tube, and the cells were incubated overnight at -20 ℃. The cells were then centrifuged at 15,000 rpm for 6 min at 4 ℃, and the supernatant was analyzed by HPLC.

[0119] HPLC detection results are as follows Figure 10 and Figure 11As shown, at a drug concentration of 100 μM, the signal of the intracellular precursor compound IMeTAGalAc-Au-PA is very low, indicating that most of the precursor may be converted into active gold and bound to thiol proteins after enzymatic cleavage. Figure 10 In addition, esterase and β-galactosidase cleavage products were also detected. The esterase cleavage product gradually increased with time, while the β-galactosidase cleavage product content remained basically unchanged with time. Under the condition of a drug concentration of 300 μM, the content of intracellular raw material IMeTAGalAc-Au-PA was observed to increase with time. Figure 11 This indicates that the raw material IMeTAGalAc-Au-PA is continuously taken up by the cells. Furthermore, a similar phenomenon to that observed at 100 μM was observed: the content of the β-galactosidase cleavage product IMeTAEtOH-Au-PA remained essentially unchanged over time. These phenomena are consistent with previous inferences: the hydrophilicity of the cleaved product IMeTAEtOH-Au-PA decreases compared to the uncleaved IMeTAGal-Au-PA, making it more susceptible to ligand exchange. However, ligand exchange requires not only a decrease in hydrophilicity but also a certain concentration threshold for the compounds to interact and promote ligand exchange.

[0120] Based on the above analysis, it is speculated that the compound IMeTAGalAc-Au-PA undergoes the following process within the cell: After entering the cell, it is first cleaved by a high concentration of estradiol to generate IMeTAGal-Au-PA. IMeTAGal-Au-PA is further cleaved by β-galactosidase, which is highly expressed in leukemia cells K562, to generate IMeTAEtOH-Au-PA. Since IMeTAEtOH-Au-PA has a lower hydrophilicity than IMeTAGal-Au-PA, when the concentration of IMeTAEtOH-Au-PA generated by cleavage accumulates to a certain threshold, interactions between molecules promote ligand exchange to generate active gold, thereby exerting an anti-leukemia effect.

[0121] Example 7: Experiment on the inhibitory effect on leukemia cells This test investigated the relationship between the inhibitory activity of compound IMeTAGalAc-Au-PA on several leukemia cells and solid tumor cells (THP-1, K562, MOLM-13, Jurkat, C1498, PC9, and SKOV3 cells) and the expression level of GLB1 in these cells. The specific steps are as follows: Taking suspension cells as an example, 10,000 cells were seeded into 96-well plates. After seeding, culture medium containing different concentrations of the compound IMeTAGalAc-Au-PA or the compound IMeTAEtOH-Au-PA was added. After incubation at 37 ℃ for 72 h in a CO2 incubator, 10 μL of CCK8 solution was added to each well, and the plates were incubated for another 2-3 h. The 96-well plates were then removed, and the absorbance at 450 nm was measured using a microplate reader to calculate the IC50 value of the compounds.

[0122] Table 2: IC50 of compounds against leukemia cells

[0123] The results above show that the enzyme digestion product IMeTAEtOH-Au-PA has significant inhibitory activity against various cancer cells. Furthermore, the inhibitory activity of IMeTAGalAc-Au-PA against several leukemia cell lines is correlated with the expression level of GLB1 (the gene encoding β-galactosidase). The GLB1 expression levels are THP-1 > K562 > MOLM-13, indicating that the inhibitory activity of the compound IMeTAGalAc-Au-PA against these three cell types is THP-1 > K562 > MOLM-13.

[0124] In summary, this invention provides an NHC-gold(I)-alkynyl complex, its preparation method, and its applications. The NHC-gold(I)-alkynyl complex of this invention exhibits solubility-related aggregation-induced ligand exchange properties; that is, under mixed solvent conditions with a low proportion of organic solvents, the complex does not dissolve well but can be dispersed well (i.e., it is within a solubility threshold). Under these conditions, ligand exchange can occur between the complex molecules through interactions to generate active gold species. Furthermore, this invention... 1 1H-NMR confirmed that the NHC-gold(I)-alkynyl complex can generate two products, NHC-Au-NHC and PA-Au-PA, through ligand exchange. Furthermore, fluorescence experiments confirmed that PA-Au-PA generated by the solubility-related aggregation-induced ligand exchange process is active gold.

[0125] Furthermore, this invention designs two NHC-Au-alkynyl complexes that can be used in click chemistry reactions, which can enhance the inhibitory activity against tumor cells when used in combination with modified trans-cyclooctyne. The compounds IMeTAGalAc-Au-PA and IMeTAGal-Au-PA of this invention also exhibit excellent tumor cell inhibitory activity. Compound IMeTAGalAc-Au-PA can be progressively cleaved by esterases and β-galactosidases, and the solubility (hydrophilicity) of the products before and after cleavage decreases compared to the complex itself before cleavage. After aggregation, reaching a certain concentration threshold, it can generate active gold species with thiol reactivity through ligand exchange. Furthermore, tumor cell inhibition experiments revealed that the compound IMeTAGalAc-Au-PA exhibits strong killing effects on various tumor cells, including leukemia cells. Cell experiments also showed that the inhibitory activity of the compound IMeTAGalAc-Au-PA on cells is positively correlated with the expression level of GLB1 (the gene encoding β-galactosidase). That is, the higher the expression level of β-galactosidase, the smaller the IC50 value of the compound IMeTAGalAc-Au-PA for that cell line. It has great application potential in the treatment of cancers with high expression of β-galactosidase.

[0126] The embodiments of the present invention have been described in detail above. However, the present invention is not limited to the above embodiments. Within the scope of knowledge possessed by those skilled in the art, various changes can be made without departing from the spirit of the present invention. Furthermore, the embodiments of the present invention and the features thereof can be combined with each other unless otherwise specified.

Claims

1. The NHC-gold(I)-alkynyl complex or a pharmaceutically acceptable salt thereof as shown in any one of Formulas I-7 to I-9: 、 、 。 2. The NHC-gold(I)-alkynyl complex or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that, Pharmaceutically acceptable salts of the NHC-gold(I)-alkynyl complex include any one of the following: hydrochloride, nitrate, acetate, methanesulfonate, phosphate, citrate, fumarate, sulfate, succinate, tartrate, citrate, hydrobromide, hydroiodide, lactate, benzylsulfonate, cinnamate, salicylate, malonate, glutarate, and malate.

3. A method for preparing NHC-gold(I)-acetylenic complexes as shown in Formulas I-7, characterized in that, Includes the following steps: In a solvent, the compound shown in Formula II is mixed with the compound shown in Formula III, and the mixture is reacted to obtain the product. The structural formula of the NHC-gold(I)-alkynyl complex represented by Formula I-7 is as follows: ； The structural formulas of the compound shown in Formula II and the compound shown in Formula III are as follows: 。 4. A pharmaceutical composition, characterized in that, The therapeutically effective amount of the NHC-gold(I)-alkynyl complex or a pharmaceutically acceptable salt thereof as described in any one of formulas I-7 to I-9 of claim 1 or 2.

5. The pharmaceutical composition according to claim 4, characterized in that, It also includes compounds BCN-OOct and / or BCN-OAc, wherein the chemical structural formulas of BCN-OOct and BCN-OAc are as follows: 。 6. The use of the NHC-gold(I)-alkynyl complex or a pharmaceutically acceptable salt thereof as described in any one of Formulas I-7 to I-9 as claimed in claim 1 or 2 in the preparation of an antitumor drug, wherein the tumor is selected from leukemia, lung cancer, colon cancer or melanoma.

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