NHC-gold (I)-alkynyl complex as well as preparation method and application thereof

Abstract

The invention discloses an NHC-gold (I)-alkynyl complex as well as a preparation method and application thereof. The structural general formula of the NHC-gold (I)-alkynyl complex is shown as a formula I: # imgabs0, in the formula I, R1 is selected from any one of hydrogen, substituted or unsubstituted C1-C10 alkyl, # imgabs1 #, # imgabs2 # and # imgabs3 #, and R2 is selected from any one of hydrogen and substituted or unsubstituted C1-C10 alkyl; r3 is selected from any one of hydrogen, a halogen atom and a substituted or unsubstituted six-membered heterocyclic ring, wherein a substituted group is a nitrogen atom or methyl; 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 disclosed by the invention can generate active gold through aggregation-induced ligand exchange reaction under the condition that no exogenous catalyst participates in, and the active gold has excellent catalytic activity and sulfydryl reaction activity and can be used for anti-tumor treatment.

Description

Technical Field

[0001] The present invention relates to the technical field of compounds, and in particular to an NHC-gold (I)-alkynyl complex and a preparation method and application thereof. Background Art

[0002] Gold is a common precious metal. Due to its excellent chemical and biological properties, 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 shrinks and the 5d orbital expands, making the gold ion have strong π acidity. Therefore, gold ions can be used as mild and efficient catalysts for activating carbon-carbon unsaturated bonds. Using the catalytic properties of gold, the luminescence of activated alkyne-catalyzed probes can be used to detect the generation of active gold species.

[0003] In the related technology, gold complexes can bind to biomacromolecules including enzymes and DNA, so gold compounds are deeply studied as anticancer and anti-rheumatic drugs. Gold drugs have an important impact on human life and health. Due to the sulfur-philic nature of gold, the target of gold drugs such as auranofin is a highly expressed enzyme in tumor cells-TrxR (thioredoxin reductase). However, in actual applications, it is 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, it will bind to gold and cause off-target effects. In order to solve the problem that gold complexes are inactivated by non-target proteins before reaching the tumor target, many researchers have designed gold complexes in the form of prodrugs to make them stably exist in non-tumor tissues. When entering tumor cells, the anti-tumor effect of gold complexes is activated by external stimulation (such as light, heat, ultrasound, etc.) or by palladium-catalyzed bioorthogonal metal transfer. However, these methods have certain limitations, such as the limited penetration of light, and palladium can also be inactivated by high concentrations of thiol species under physiological conditions. Therefore, a large amount of palladium catalyst is required to achieve the ideal anti-tumor effect.

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

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

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

[0007] The invention also provides a method for preparing the NHC-gold (I)-alkynyl complex.

[0008] The present invention also proposes the use of the NHC-gold (I)-alkynyl complex shown in formula I or its pharmaceutically acceptable salt, ester, solvate, prodrug in the preparation of active gold.

[0009] The present invention also provides a use of the NHC-gold (I)-alkynyl complex as shown in formula I or a pharmaceutically acceptable salt, ester, solvate, or prodrug thereof in the preparation of an anti-tumor drug.

[0010] In a first aspect of the present invention, there is provided an NHC-gold (I)-alkynyl complex as shown in formula I or a pharmaceutically acceptable salt, ester, solvate, or prodrug thereof, ; Wherein, R 1 is selected from hydrogen, substituted or unsubstituted C1~C10 alkyl, , , Any one of the following, wherein the substituted group is an azide group; The R 2 Any one selected from hydrogen, substituted or unsubstituted C1~C10 alkyl; The R 3 Any one selected from hydrogen, a halogen atom, a substituted or unsubstituted six-membered heterocyclic ring, wherein the substituted group is a nitrogen atom or a methyl group; The R 4 and the R 5 are independently hydrogen, or said R 4 and the R 5 Connect to form an unsaturated six-membered ring.

[0011] The NHC-gold (I)-alkynyl complex according to the embodiment of the present invention has at least the following beneficial effects: The NHC-gold (I)-alkynyl complex of the present invention can undergo an aggregation-induced ligand exchange reaction. The present invention proves that the NHC-gold (I)-alkynyl complex has the property of solubility-related aggregation-induced ligand exchange by adjusting the ratio of the organic solvent and water / PBS in the mixed solvent, that is, under the mixed solvent condition of a lower ratio of organic solvent, the complex cannot be well dissolved but can be well dispersed (that is, it is in a condition of a solubility threshold). Under this condition, the complex molecules will interact with each other to exchange ligands, thereby generating active gold species. In addition, the active gold generated after the reaction of the NHC-gold (I)-alkynyl complex of the present invention has excellent catalytic activity and thiol reaction activity, can inhibit tumor cell proliferation, and can be used for anti-tumor treatment.

[0012] In some embodiments of the present invention, R 3 Any one selected from hydrogen, a halogen atom, and a methyl tetrazine group.

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

[0014] .

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

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

[0017] The compound IMeTAGalAc-Au-PA or IMeTAGalAc-Au-PA of the present invention can react with esterase and / or β-galactosidase expressed in tumor cells. Specifically, the compound IMeTAGalAc-Au-PA can be gradually cleaved by esterase and β-galactosidase, the compound IMeTAGal-Au-PA can be cleaved by β-galactosidase, and the solubility (hydrophilicity) of the product before and after cleavage is reduced relative to the complex itself before cleavage, which helps to improve the aggregation effect, trigger ligand exchange reaction, and then generate active gold for inhibiting 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, toluenesulfonate, cinnamate, salicylate, malonate, glutarate, and malate.

[0019] The second aspect of the present invention provides a method for preparing the NHC-gold (I)-alkynyl complex as shown in formula I-7, comprising the following steps: In a solvent, the compound represented by formula II and the compound represented by formula III are mixed and reacted to obtain a compound represented by formula I-7 as shown below: ; The structural formulas of the compound represented by formula II and the compound represented by formula III are as follows: .

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

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

[0022] The pharmaceutical composition according to the embodiment 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 in the absence of a 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 the compound 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 is used in combination with the compound BCN-OOct and / or BCN-OAc, which can significantly improve the killing power to tumor cells and has a positive effect on improving the tumor treatment effect.

[0025] In some embodiments of the present invention, the pharmaceutical composition further comprises a pharmaceutically acceptable excipient.

[0026] In some embodiments of the present invention, the pharmaceutically acceptable excipients include at least one of a solvent, an excipient, a diluent, a binder, a disintegrant, a dispersant, a flavoring agent, a suspending agent, a surfactant, an isotonic agent, a thickener, an emulsifier, a preservative, a glidant, and a lubricant.

[0027] In some embodiments of the present invention, the dosage form of the drug is any one of tablets, capsules, granules, injections, powder injections, 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 comprises at least one of starch and sucrose.

[0030] In some embodiments of the present invention, the binder includes at least one of a cellulose derivative, alginate, gelatin, and polyvinyl pyrrolidone.

[0031] In some embodiments of the invention, the humectant comprises glycerin.

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

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

[0034] In some embodiments of the invention, the surfactant comprises cetyl alcohol.

[0035] In some embodiments of the present invention, the adsorption carrier includes at least one of kaolin and bentonite.

[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 any conventional dosage form in the art.

[0038] In some embodiments of the present invention, the dosage form of the pharmaceutical composition is in the form of solid, semisolid or liquid, and can be an aqueous solution, non-aqueous solution or 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, pellets, dry extracts, injections, and infusions.

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

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

[0042] The application of the embodiment of the present invention has at least the following beneficial effects: the NHC-gold (I)-alkynyl complex of the present invention can undergo ligand exchange reaction through aggregation to generate active gold, and the active gold has excellent catalytic activity and thiol reaction activity, and can be used for anti-tumor treatment. In some embodiments of the present invention, when the NHC-gold (I)-alkynyl complex is a compound represented by Formula I-1 to Formula I-6 or Formula I-9, the method for preparing active gold comprises: The NHC-gold (I)-alkynyl complex is placed in a solvent and / or a buffer for reaction to obtain the complex.

[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)-alkynyl complex.

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

[0045] In some embodiments of the invention, the solution comprises a phosphate buffer.

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

[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 comprises: The NHC-gold (I)-alkynyl complex is mixed with esterase and β-galactosidase in a solvent and / or buffer, and then reacted and purified to obtain the complex.

[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 comprises: The NHC-gold (I)-alkynyl complex (IMeTAGal-Au-PA) is mixed with β-galactosidase in a solvent and / or buffer, and then reacted and purified to obtain the complex.

[0049] The fifth aspect of the present invention provides the use of the NHC-gold (I)-alkynyl complex as shown in formula I or a pharmaceutically acceptable salt, ester, solvate, or prodrug thereof as described in any one of the first aspect in the preparation of an anti-tumor drug.

[0050] According to the application of the embodiment of the present invention, at least the following beneficial effects are achieved: the NHC-gold (I)-alkynyl complex of the present invention can generate NHC-Au-NHC and active gold products through ligand exchange, which has a strong killing effect on a variety of tumor cells including leukemia cells, and when it is combined with modified trans-cyclooctyne, the inhibitory activity on tumor cells is further enhanced. In addition, the present invention also found through cell experiments that the inhibitory activity of the compound IMeTAGalAc-Au-PA on cells is positively correlated with the expression level of GLB1 (gene encoding β-galactosidase) of the cells, that is, the higher the expression level of β-galactosidase, the smaller the IC50 value of the compound IMeTAGalAc-Au-PA for the cell line, and has a good application prospect in 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] In addition, it can be understood that the term "substituted or unsubstituted" in the present invention refers to two situations in which one or more hydrogen atoms on the substituted group can be "substituted" or "unsubstituted" by one or more substituents. The number of substitutions ranges from single substitution to the maximum number of substitutions.

[0054] Other features and advantages of the present invention will be set forth in the description which follows. BRIEF DESCRIPTION OF THE DRAWINGS

[0055] The present invention will be further described below with reference to the accompanying drawings and embodiments, wherein: Figure 1 is the HRMS chart of compound 11 of the present invention; Figure 2 HRMS chart of the compound IMeTAGalAc-Au-PA of the present invention; Figure 3 HRMS chart of the compound IMeTAGal-Au-PA of the present invention; Figure 4 HRMS chart of the compound IMeTAEtOH-Au-PA of the present invention; Figure 5 The present invention is a method for generating NHC-Au-NHC and PA-Au-PA by ligand exchange of NHC-gold (I)-alkynyl complex 1a. 1 H-NMR spectrum; Figure 6 Schematic diagram of the reaction mechanism of the NHC-gold (I)-alkynyl complex aggregation-induced ligand exchange to produce active gold; Figure 7 A diagram showing the luminescence of Probe-1 catalyzed by active gold produced by solubility-dependent aggregation-induced ligand exchange of the NHC-gold (I)-alkynyl complex of the present invention; Figure 8 It is a schematic diagram of the step-by-step enzymatic cleavage process of the β-galactosidase-responsive gold (I) complex IMeTAGalAc-Au-PA of the present invention, and a HPLC characterization diagram of the enzymatic cleavage process at the extracellular level; Fig. 9 HPLC characterization diagram of the reaction degree of IMeTAGal-Au-PA and IMeTAEtOH-Au-PA after the enzymatic cleavage of the β-galactosidase-responsive gold (I) complex IMeTAGalAc-Au-PA of the present invention with GSH under the same solvent conditions; Fig.10 HPLC characterization diagram of the enzymatic cleavage process of the β-galactosidase-responsive gold (I) complex IMeTAGalAc-Au-PA (100 μM) of the present invention at the intracellular level; Fig.11 HPLC characterization chart of the enzymatic cleavage process of the β-galactosidase-responsive gold (I) complex IMeTAGalAc-Au-PA (300 μM) of the present invention at the intracellular level. DETAILED DESCRIPTION

[0056] The following will be combined with the embodiments to clearly and completely describe the concept of the present invention and the technical effects produced, so as to fully understand the purpose, characteristics and effects of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, other embodiments obtained by those skilled in the art without creative work are all within the scope of protection of the present invention.

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

[0058] When a numerical range is disclosed herein, the above range is considered to be continuous and includes the minimum and maximum values ​​of the range, as well as every value between such minimum and maximum values. Further, when a range refers to an integer, every integer between the minimum and maximum values ​​of the range is included. In addition, when multiple ranges are provided to describe features or characteristics, the ranges can be combined. In other words, unless otherwise indicated, all ranges disclosed herein should be understood to include any and all subranges included therein.

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

[0060] In the description of the present invention, the description with reference to the terms "one embodiment", "some embodiments", "illustrative embodiments", "examples", "specific examples", or "some examples" means that the specific features, structures, materials or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the schematic representation of the above terms does not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described may be combined in any one or more embodiments or examples in a suitable manner.

[0061] In the examples of the present invention, the RPMI 1640 culture medium and fetal bovine serum (FBS) used in the cell experiments were from Gibco, USA, and penicillin and streptomycin were purchased from HyClone, USA. Unless otherwise specified, all chemicals were from commercial sources, mainly from Bidex Pharmaceuticals and Anergy Chemicals.

[0062] The cells used in the experiment include 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 are from the research group of Zhang Xiaolei of the School of Pharmacy of Sun Yat-sen University, and the remaining cells are from the American Type Culture Collection (ATCC). In the embodiments or tests of the present invention, the cancer cells are cultured in the following manner: Take the culture flask with the cancer cells required for the experiment, collect the cell suspension and centrifuge, discard the original culture medium, add 2 mL of fresh culture medium and mix by pipetting. Take 10 μL of cell suspension to the cell counting plate and count; spread on a 96-well plate, seed 10,000 cells per well, calculate the required cell volume and cell well volume for 3 replicates of each compound, calculate the required cell suspension volume by cell counting, and the required cell number of culture medium dilution, 10,000 cells per well, 100 μL of culture medium, spread on the plate, mix by pipetting, use a spray gun to seed the cells, and place in CO 2 Incubate in 37°C incubator and mark the plates.

[0063] In addition, if the specific conditions are not specified in the examples, the experiments were carried out according to conventional conditions or the conditions recommended by the manufacturer. The reagents or instruments used, if the manufacturers are not specified, are all conventional products that can be purchased from the market.

[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 compound was 1,3-dimethylimidazole chloride (0.5 mmol, 66.3 mg), DMSAuCl (0.5 mmol, 147 mg), K 2 CO 3 (1.5 mmol, 207 mg) was added into a 25 mL Schlenk tube, a magnetic stirrer was added into the tube, and then 5 mL acetone was added to dissolve the raw materials. The reaction tube was transferred into an oil bath and heated to 60 °C for 1 h. After the reaction was completed, the reaction was filtered and the filtrate was dried 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 (5eq.1 mmol) were dissolved in 15 mL MeOH and reacted at 80 °C for 1 h. The obtained solid was spin-dried, redissolved in DCM and filtered, and the solution was concentrated to ~ 1 mL and recrystallized by adding n-pentane to obtain 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 a NHC-gold (I)-alkynyl complex FPA (hereinafter referred to as compound FPA), the structural formula of which 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 (5eq. 1 mmol) were dissolved in 15 mL MeOH and reacted at 80 °C for 1 h. The solid was spin-dried, redissolved in DCM and filtered, and the solution was concentrated to ~ 1 mL and recrystallized by adding n-pentane to obtain a yellow-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 a 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 (5eq. 1 mmol) were dissolved in 15 mL MeOH and reacted at 80 °C for 1 h. The solid was spin-dried, redissolved in DCM and filtered, and the solution was concentrated to ~ 1 mL and recrystallized by adding n-pentane to obtain 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 a NHC-gold (I)-alkynyl complex Oct (hereinafter referred to as compound Oct), the structural formula of which is shown in Formula I-4: .

[0076] The preparation method of the above compound Oct is as follows: ; Compounds 1-methyl-3-octylimidazolium chloride (0.5 mmol, 115.4 mg), DMSAuCl (0.5 mmol, 147 mg), K 2 CO 3(1.5 mmol, 207 mg) was added to a 25 mL Schlenk tube, a magnetic stirrer was added to the tube, and then 5 mL acetone was added to dissolve the raw materials. The reaction tube was transferred to an oil bath and heated to 60 °C for 1 h. After the reaction was completed, the filtrate was dried and the crude product Oct-Au-Cl was obtained as an oily compound. All the crude products were added to the reaction tube, and phenylacetylene (0.5 mmol, 51 mg) and NaOMe (5eq. 2.5 mmol) were added and dissolved in 15 mL MeOH, and the reaction was carried out at 80 °C for 1 h. The solid was dried and re-dissolved in DCM and filtered. The solution was concentrated to ~ 1 mL and recrystallized by adding n-pentane to obtain 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: The synthesis methods of Compound 1 and Compound 2 are reported in reference literature. Specifically, Compound 1 was synthesized by the method described in reference literature "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 reference literature "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), K 2 CO 3 (1.5 mmol, 207 mg) was added to a 25 mL Schlenk tube, a magnetic stirrer was added to the tube, and then 5 mL of acetone was added to dissolve the raw materials, and the reaction tube was transferred to an oil bath and heated to 60 ° C, and the reaction was carried out for 1 h. After the reaction was completed, the filtrate was filtered and the filtrate was dried to obtain the crude product 3. All the crude product 3 was added to a 25 mL Schlenk tube, phenylacetylene (0.5 mmol, 51 mg) and sodium methoxide (1 mmol, 51 mg) were added, and then 5 mL of methanol was added. The reaction tube was transferred to an oil bath and heated to 80 ° C. The reaction was carried out for 1 h. After the reaction was completed, the filtrate was dried and redissolved. After filtration, the filtrate was recrystallized in MeOH / ether system to obtain the 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 a NHC-gold (I)-alkynyl complex Tz (hereinafter referred to as compound Tz), the structural formula of the compound is shown in formula I-6: .

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

[0085] The specific preparation process is: 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, and the reaction tube was reacted at 80 °C for 1 h. After the reaction was completed, the mixture was spin-dried, redissolved, and filtered. The filtrate was concentrated and dissolved in a small amount of DCM, and n-pentane was added for recrystallization to obtain compound Tz as a rose-red powder with a yield of 73%.

[0087] The characterization data of compound Tz are as follows: 1H 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 a NHC-gold (I)-alkynyl complex IMeTAGalAc-Au-PA (hereinafter referred to as compound IMeTAGalAc-Au-PA), the structural formula of the compound is shown in Formula I-7: .

[0089] The specific reaction process is as follows: ; The specific preparation process is: (1) Compounds 5, 6 and 7 were synthesized according to the method 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 Cell Lines [J]. Biomacromolecules, 2015, 16(7): 1948-57”; Compound 7 was prepared according to the reference “HOOGBOOM J, SWAGER TM. 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 refluxed in an oil bath for 24 h. After the reaction, the solvent was dried and the crude product was recrystallized from MeOH / ether to obtain compound 8 as brown-yellow crystals with a yield of 46%.

[0091] The characterization data of 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). 13 C 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 K 2 CO 3 (0.5 mmol, 69 mg) was added into a 25 mL Schlenk tube, a magnetic stirrer was added into the tube, and then 5 mL acetone was added to dissolve the raw material. The reaction tube was transferred to an oil bath and heated to 60 °C for 3 h. After the reaction was completed, the filtrate was filtered, and a small amount of DCM was added to redissolve the filtrate after drying. The compound 9 was recrystallized from n-pentane as a white powder with a yield of 68%.

[0093] The characterization data of 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, and phenylacetylene (0.187 mmol, 19 mg) and sodium methoxide (0.34 mmol, 19 mg) were added, and then 5 mL of methanol was added. The reaction tube was transferred to an oil bath and heated to 80 °C for 1 h. After the reaction, the mixture was spin-dried, redissolved and filtered, and the filtrate was spin-dried to obtain a crude product of compound 10. Compound 10 was completely dissolved in DMF, and an appropriate amount of CsF was added to react at room temperature for 2 h. After the reaction, the mixture was spin-dried, redissolved in DCM and filtered, and the filtrate was collected, concentrated, and recrystallized from DCM / n-pentane to obtain compound 11 as a yellow-white powder with a yield of 56%.

[0095] The HRMS pattern of compound 11 is shown in Figure 1 As shown, its characterization data are as follows: 1 H 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 placed in 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 dried and 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] The HRMS image of compound IMeTAGalAc-Au-PA is as follows Figure 2 As shown, its characterization data are as follows: 1 H 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 a NHC-gold (I)-alkynyl complex IMeTAGal-Au-PA (hereinafter referred to as compound IMeTAGal-Au-PA), the structural formula of the compound is shown in Formula I-8: .

[0098] The reaction process is as follows: .

[0099] The specific preparation method includes: 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 bottle, 5 mL of methanol was added to dissolve the raw materials, and after reacting at room temperature for 3 h, 1 M hydrochloric acid was added to neutralize the pH to neutral, the solvent was spin-dried, and the product 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 image of compound IMeTAGal-Au-PA is shown below: Figure 3 As shown, its characterization data are as follows: HRMS (Exact mass667.17, found 668.1773 (+1)).

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

[0102] The reaction process is as follows: .

[0103] The specific preparation method includes: Azideethanol 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 placed in a 25 mL Schlenk tube under nitrogen atmosphere, and 5 mL of acetonitrile was added to dissolve the raw materials. After reacting at room temperature for 24 h, the solvent was dried by spin drying and then filtered through Al 2 O 3 The compound IMeTAGalAc-Au-PA was obtained by column (DCM / MeOH=150:1) as a yellow-white powder with a yield of 68%.

[0104] The HRMS image of compound IMeTAEtOH-Au-PA is as follows Figure 4 As shown, its characterization data are 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] Test Example 1: Verification of the ligand exchange reaction of NHC-gold(I)-alkynyl complex 1a This test example takes the NHC-gold (I)-alkynyl complex 1a of Example 1 as an example to verify whether the NHC-gold (I)-alkynyl complex can undergo ligand exchange, and the products generated by the ligand exchange. The specific process is as follows: Weigh 2.5 nmol of compound 1a and dissolve it in 200 μL DMSO, add 200 μL PBS, and shake at 37°C for 12 hours. After the reaction, add 400 μL DMSO to dissolve the insoluble matter in the EP tube, add 200 μL d6-DMSO, take 250 μL of liquid from each EP tube, add 300 μL DMSO to dilute, and use 1H-NMR to detect the difference between the nuclear magnetic resonance hydrogen spectra before and after the reaction.

[0106] Test results such as Figure 5 As shown, the 1H-NMR spectrum shows that the nuclear magnetic resonance hydrogen spectrum of compound 1a before and after the reaction is very different. The solvent conditions used in the reaction can ensure that compound 1a is in a poorly soluble state. In this state, it can be inferred that the molecules of compound 1a will interact with each other, leading to the ligand exchange process, and then generating the ligand exchange products NHC-Au-NHC and PA-Au-PA, whose structural formulas are as follows: .

[0107] Test Example 2: Verification of the catalytic effect of active gold in the product This test example studies whether the active gold generated by the above-mentioned NHC-gold (I)-alkynyl complexes (such as compound 1a, compound FPA, compound Nap, compound Oct) after ligand exchange can have a catalytic effect, such as catalyzing the luminescence of the coumarin precursor probe Probe-1 (a probe that can be catalyzed by gold and can be used to detect active gold species). The specific experimental process is as follows: Take 7 EP tubes from each group, add 1 mL of MeCN / PBS mixed solvent (the ratio of MeCN in the solvent is: 100%, 80%, 60%, 40%, 20%, 10%, 0%), then add Au 20 μM and Probe-1 100 μM, and place the EP tube at 37 °C for shaking reaction for 24 h. The reaction mechanism diagram is shown in the figure Figure 6 shown.

[0108] Test results such as Figure 7 As shown, NHC-gold (I)-alkynyl complex 1a, FPA, Nap and Oct can catalyze the luminescence of Probe-1 in a mixed solvent with a low MeCN content.

[0109] In addition, based on the above detection example 1, it can be seen that the 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, but PA-Au-PA can, indicating that the ligand exchange of compound 1a can produce active gold species PA-Au-PA. Combined with the above experiments, it can be seen that compounds FPA, Nap and Oct can achieve comparable technical effects.

[0110] Test Example 3: Detection of the inhibitory effect of compound N3 or Tz on tumor cells This test example tests the inhibitory effect of compound N3 or compound Tz combined with modified trans-cyclooctyne on various tumor cells. The specific steps are as follows: 5000 tumor cells were plated in each well of a 96-well plate. After 24 h, the trans-cyclooctyne concentration was fixed at 25 μM, and medium containing different concentrations of compound N3 or Tz was added. 2 After culturing in an incubator at 37 °C for 48 h, 15 μL of MTT solution was added to each well, and then the plate was incubated in an incubator for another 3 h. The 96-well plate was taken out, and 100 μL of DMSO was added to each well to dissolve the generated formazan. The absorbance at 490 nm was detected with an enzyme-labeled instrument to calculate the IC50 value of the compound.

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

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

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

[0114] Detection Example 4: Enzyme cleavage reaction of detection compound IMeTAGalAc-Au-PA This test example tests whether the compound IMeTAGalAc-Au-PA can be cleaved step by step by esterase and β-galactosidase. The specific steps are as follows: The compound IMeTAGalAc-Au-PA 200 μM was added to 1 mL PBS, and Esterase 10 U / mL was added first to react at 37 °C for 3 h, and then β-Gal 5 U / mL was added and the reaction was continued at 37 °C overnight. After the reaction, the protein was precipitated with acetone and the supernatant was subjected to HPLC.

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

[0116] Detection Example 5: Detection of the active gold thiol reaction activity of the enzyme cleavage product This test example detects the product of the enzymatic cleavage of the compound IMeTAGalAc-Au-PA, that is, the reaction degree of the compound IMeTAGal-Au-PA and the compound IMeTAEtOH-Au-PA with glutathione (GSH) under the same solvent conditions. The specific steps are as follows: 200 μM compound IMeTAGal-Au-PA or compound IMeTAEtOH-Au-PA was added to 500 μL of DMF / PBS mixed solvent, with the DMF ratio of 10%, 2% or 90% (including the volume of the added Au stock solution), containing 10 mM GSH (equivalent triethylamine to neutralize pH), reacted at 37 °C for 24 h, added 500 μL DMF to dissolve, and measured by HPLC.

[0117] HPLC test results are as follows Fig. 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, that is, the proportion of compound IMeTAEtOH-Au-PA reacting with GSH is greater than that of compound IMeTAGal-Au-PA, indicating that the hydrophilicity of the product IMeTAEtOH-Au-PA after enzyme cleavage is reduced compared with IMeTAGal-Au-PA before enzyme cleavage, and it is more likely to undergo ligand exchange to generate active gold PA-Au-PA. The generated active gold has thiol reaction activity. It can be seen that compared with compound IMeTAGal-Au-PA, compound IMeTAEtOH-Au-PA has a higher degree of reaction with GSH.

[0118] Test Example 6: Cell Experiment This test example detected the content of the compound IMeTAGalAc-Au-PA and its enzyme cleavage product in K562 cells as the incubation time increased. The specific steps are as follows: K562 cells were inoculated in 10 cm cell culture dishes, and the compound IMeTAGalAc-Au-PA was administered at 100 μM and incubated for 2 h, 4 h, 8 h, and 12 h, or 300 μM and incubated for 1 h, 2 h, 4 h, and 8 h. The cells were then collected by centrifugation, washed three times with PBS, added with 200 μL of ice water, and repeatedly frozen and thawed at -80 ℃ for three times to lyse the cells. 600 μL of ice acetone was added to each EP tube, precipitated at -20 ℃ overnight, centrifuged at 4 ℃ and 15000 rpm for 6 min, and the supernatant was measured by HPLC.

[0119] HPLC test results are as follows Fig.10 and Fig.11 As shown in the figure, under the condition of 100 μM drug concentration, the signal of intracellular raw material compound IMeTAGalAc-Au-PA is very low, indicating that most of the raw materials may be converted into active gold after enzyme cleavage and bind to thiol protein ( Fig.10 In addition, the cleavage products of Esterase and β-galactosidase can also be detected. The cleavage products of Esterase gradually increase with time, while the content of β-galactosidase cleavage products basically does not change with time; under the condition of a drug concentration of 300 μM, it can be observed that the content of intracellular raw material IMeTAGalAc-Au-PA increases with time ( Fig.11 ), indicating that the raw material IMeTAGalAc-Au-PA is continuously taken up by cells. In addition, similar phenomena to those observed when 100 μM was administered, the content of IMeTAEtOH-Au-PA, the enzymatic cleavage product of β-galactosidase, basically did not change over time. The above phenomenon is consistent with the previous inference: the hydrophilicity of the product IMeTAEtOH-Au-PA after enzymatic cleavage is reduced compared to IMeTAGal-Au-PA before enzymatic cleavage, which makes ligand exchange more likely to occur. The conditions required for ligand exchange are not only a decrease in hydrophilicity, but also the need for the concentration of the compound to reach a certain threshold so that the compound molecules can interact with each other and promote the ligand exchange process.

[0120] Based on the above analysis, it is speculated that the compound IMeTAGalAc-Au-PA undergoes the following process in the cell: after entering the cell, it is first cleaved by a high concentration of Esterase to generate the compound IMeTAGal-Au-PA, and IMeTAGal-Au-PA is further cleaved by β-galactosidase highly expressed in leukemia cells K562 to generate IMeTAEtOH-Au-PA. Due to the decrease in the hydrophilicity of IMeTAEtOH-Au-PA relative to IMeTAGal-Au-PA, when the concentration of IMeTAEtOH-Au-PA produced by enzyme cleavage accumulates to a certain threshold, the molecules interact with each other to promote ligand exchange to generate active gold, thereby exerting an anti-leukemia effect.

[0121] Test Example 7: Leukemia Cell Inhibition Effect Experiment This test example detected the relationship between the inhibitory activity of the compound IMeTAGalAc-Au-PA on several leukemia cells and solid tumor cells (THP-1, K562, MOLM-13, Jurkat, C1498, PC9, SKOV3 cells) and the expression level of GLB1 in cells. The specific steps are as follows: Taking suspension cells as an example, 10,000 cells were plated in each well of a 96-well plate, and culture medium containing different concentrations of compound IMeTAGalAc-Au-PA or compound IMeTAEtOH-Au-PA was added after the plating was completed. 2 After culturing in an incubator at 37 °C for 72 h, 10 μL of CCK8 solution was added to each well, and then the plate was incubated in the incubator for 2-3 h. The 96-well plate was taken out, and the absorbance at 450 nm was measured on a microplate reader to calculate the IC50 value of the compound.

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

[0123] The above results show that the enzyme cleavage product IMeTAEtOH-Au-PA has significant inhibitory activity against a variety of cancer cells. In addition, the inhibitory activity of IMeTAGalAc-Au-PA against several leukemia cells is related to the expression of GLB1 (gene encoding β-galactosidase) in the cells. The expression of GLB1 is THP-1>K562>MOLM-13, and the inhibitory activity of the compound IMeTAGalAc-Au-PA against these three cells is THP-1>K562>MOLM-13.

[0124] In summary, the present invention provides an NHC-gold (I)-alkynyl complex and its preparation method and application. The NHC-gold (I)-alkynyl complex of the present invention has the property of solubility-related aggregation-induced ligand exchange, that is, under the condition of a mixed solvent with a lower proportion of organic solvent, the complex cannot be well dissolved but can be well dispersed (that is, it is in a solubility threshold condition). Under this condition, the complex molecules can interact with each other to exchange ligands to produce active gold species. 1 H-NMR demonstrated that the NHC-gold(I)-alkynyl complex could generate two products, NHC-Au-NHC and PA-Au-PA, through ligand exchange, and fluorescence experiments demonstrated that the PA-Au-PA produced by the solubility-dependent aggregation-induced ligand exchange process was active gold.

[0125] In addition, the present invention designs two NHC-Au-alkynyl complexes that can be used for click chemistry reactions, which can be used in combination with modified trans-cyclooctyne to enhance the inhibitory activity against tumor cells. The compounds IMeTAGalAc-Au-PA and IMeTAGal-Au-PA of the present invention also have excellent tumor cell inhibitory activity, wherein the compound IMeTAGalAc-Au-PA can be gradually cleaved by esterase and β-galactosidase, and the solubility (hydrophilicity) of the products before and after cleavage is reduced relative to the complex itself before cleavage, and after aggregation, when a certain concentration threshold is reached, active gold species with thiol reaction activity can be generated through ligand exchange. Furthermore, through tumor cell inhibition experiments, it was found that the compound IMeTAGalAc-Au-PA has a strong killing effect on a variety of tumor cells including leukemia cells, and through cell experiments, it was also found that the inhibitory activity of the compound IMeTAGalAc-Au-PA on cells was positively correlated with the expression level of GLB1 (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 the cell line, which has a good application prospect in the treatment of cancers with high expression of β-galactosidase.

[0126] The above is a detailed description of the embodiments of the present invention, but the present invention is not limited to the above embodiments. Various changes can be made within the knowledge of ordinary technicians in the relevant technical field without departing from the purpose of the present invention. In addition, the embodiments of the present invention and the features in the embodiments can be combined with each other without conflict.

Claims

1. An NHC-gold (I)-alkynyl complex as shown in formula I or a pharmaceutically acceptable salt, ester, solvate, or prodrug thereof, ； in, In the formula I, R1 is selected from hydrogen, substituted or unsubstituted C1~C10 alkyl, , , Any one of the following, wherein the substituted group is an azide group; R2 is selected from any one of hydrogen, substituted or unsubstituted C1~C10 alkyl; The R3 is selected from any one of hydrogen, halogen atoms, substituted or unsubstituted six-membered heterocyclic rings, wherein the substituted group is a nitrogen atom or a methyl group; The R4 and the R5 are independently hydrogen, or the R4 and the R5 are connected to form an unsaturated six-membered ring.

2. The NHC-gold (I)-alkynyl complex of formula I or a pharmaceutically acceptable salt, ester, solvate or prodrug thereof according to claim 1, characterized in that: In the formula I, R3 is selected from any one of hydrogen, halogen atom and methyl tetrazine group.

3. The NHC-gold (I)-alkynyl complex of formula I or a pharmaceutically acceptable salt, ester, solvate or prodrug thereof according to claim 1, characterized in that: The NHC-gold (I)-alkynyl complex is selected from any one of the compounds of formula I-1 to formula I-9, 。 4. The NHC-gold (I)-alkynyl complex of formula I or a pharmaceutically acceptable salt, ester, solvate or prodrug thereof according to claim 1, characterized in that: 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, toluenesulfonate, cinnamate, salicylate, malonate, glutarate and malate.

5. A method for preparing the NHC-gold (I)-alkynyl complex as shown in formula I-7, characterized in that: The following steps are involved: In a solvent, the compound represented by formula II and the compound represented by formula III are mixed and reacted to obtain the compound; wherein the structural formulas of the compound represented by formula II and the compound represented by formula III are as follows: 。 6. A pharmaceutical composition, characterized in that A method comprising a therapeutically effective amount of the NHC-gold (I)-alkynyl complex as shown in formula I according to any one of claims 1 to 4 or a pharmaceutically acceptable salt, ester, solvate, or prodrug thereof.

7. The pharmaceutical composition according to claim 6, characterized in that It also contains compounds BCN-OOct and / or BCN-OAc, wherein the chemical structural formulas of the BCN-OOct and BCN-OAc are as follows: 。 8. Use of the NHC-gold (I)-alkynyl complex of formula I or a pharmaceutically acceptable salt, ester, solvate or prodrug thereof as claimed in any one of claims 1 to 4 in the preparation of active gold.

9. Use of the NHC-gold (I)-alkynyl complex of formula I or a pharmaceutically acceptable salt, ester, solvate or prodrug thereof as claimed in any one of claims 1 to 4 in the preparation of an anti-tumor drug.

10. The use according to claim 9, characterized in that: The tumor is a tumor capable of expressing β-galactosidase and / or esterase.

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