A composition containing aurothioglucose and polysulfide and its use

Abstract

The application discloses a composition containing auro-thio-logic acid and polysulfide and application thereof, and belongs to the technical field of biological medicines, wherein the composition comprises auro-thio-logic acid and polysulfide. The application solves the defect that auro-thio-logic acid cannot efficiently inhibit cancer cell proliferation under normal physiological conditions by combining auro-thio-logic acid and polysulfide. When combined with polysulfide, auro-thio-logic acid can well inhibit the growth of tumors, indicating that auro-thio-logic acid and polysulfide can be applied as an anticancer drug combination. Moreover, the absorption of auro-thio-logic acid in tumor tissues can be adjusted by adjusting the type of polysulfide, so that the composition has good selectivity in anticancer in vivo. The composition has good stability and is not easy to be oxidized by air. The drug combination can also achieve an anticancer effect by oral (intragastric administration) mode, and is not limited to intraperitoneal or intravenous injection mode. Meanwhile, the drug combination also has good antibacterial effect.

Description

Technical Field

[0001] This invention relates to the field of biomedical technology, specifically to a composition containing aurinophene and polysulfide compounds and its application. Background Technology

[0002] Aurinophene is an oral antirheumatic drug primarily used to treat active rheumatoid arthritis. Early studies showed that in mice inoculated with P388 acute lymphoblastic leukemia cells via intraperitoneal injection (ip), aurinophene was only effective with ip administration; other routes of administration (e.g., intravenous, subcutaneous, or oral) were ineffective. In mice inoculated with P388 acute lymphoblastic leukemia cells via other methods, aurinophene was ineffective regardless of the administration method. Subsequent studies showed that in several solid tumor models (osteosarcoma, melanoma, lung cancer, colon cancer, and breast cancer), aurinophene neither prolonged lifespan nor inhibited tumor growth. In recent years, a few studies have reported anticancer activity with aurinophene alone; however, the doses used were very high (close to lethal doses).

[0003] In vivo applications, the pharmacological activity of aurnofen is significantly weakened when used alone or through simple pharmacological combinations because aurnofen readily binds to human serum albumin in the blood. Therefore, further development of drug combinations that can enhance the anticancer activity of aurnofen is of great significance for the research and development of anticancer drugs. Summary of the Invention

[0004] The purpose of this invention is to overcome the shortcomings of the prior art and provide a composition containing aurinolone and polysulfide compounds and its application. The composition can effectively inhibit the proliferation of cancer cells and the growth of tumors, while also having good antibacterial effects.

[0005] Therefore, in a first aspect of the invention, a composition comprising aurinophene and a polysulfide compound is provided.

[0006] The prior art (CN112691196A) discloses an anticancer composition containing aurnofen and a thiol compound and its application. It further discloses that the thiol compound can effectively inhibit the binding of aurnofen to thiol groups in blood proteins in vivo, thereby enhancing the cellular uptake of aurnofen and regulating the in vivo anticancer activity of aurnofen. However, the inventors found that the thiol compound is easily oxidized by air, has insufficient stability, and can only be administered via intraperitoneal or intravenous injection, which limits its application.

[0007] While experimenting with numerous compounds, the inventors stumbled upon a discovery that aurnofen, when used in combination with polysulfide compounds, exhibited a remarkably strong tumor-suppressing effect. Under safe dosage conditions, aurnofen or polysulfide compounds alone showed poor tumor-suppressing effects; however, when used in combination, they significantly inhibited tumor growth, with polysulfide compounds significantly enhancing the antitumor effect of aurnofen.

[0008] The inventors further investigated the mechanism and discovered that auronorfen rapidly binds to human serum albumin (HSA) after entering the human body, reducing the effective uptake of gold ions and decreasing its cytotoxicity, thus preventing it from exerting an effective cancer-suppressing effect. The composition of this invention contains polysulfide compounds that can inhibit the binding of auronorfen to sulfhydryl groups in blood proteins, thereby enhancing cellular uptake of auronorfen and regulating its in vivo anticancer activity.

[0009] By utilizing the ligand exchange properties of aurinophene and its high affinity for thiol groups, the in vivo anticancer activity of aurinophene can be restored by adding specific polysulfide compounds, thus giving the composition higher anticancer activity.

[0010] As a preferred embodiment of the present invention, the polysulfide compound includes compounds with structural formulas such as Formula I, Formula II, Formula III, and Formula IV; or

[0011] Isomers, pharmaceutically acceptable hydrates, solvates, or salts of compounds represented by Formulas I, II, III, and IV;

[0012]

[0013]

[0014] In Equations I and II, n is an integer ≥ 1;

[0015] Among them, R1, R2, R3, R4, R5, and R6 are independently selected from alkyl, alkenyl, carbonyl, aryl, alkyl derivatives containing heteroatoms, alkenyl derivatives containing heteroatoms, carbonyl derivatives containing heteroatoms, or aryl derivatives containing heteroatoms.

[0016] The heteroatom is at least one of N, S, O, P, and Si.

[0017] The inventors further investigated the stability and administration method of the above-mentioned compositions and found that the presence of disulfide bonds in the compounds represented by Formulas I, II, III, and IV is key. The compounds represented by Formulas I, II, III, and IV, or their isomers, pharmaceutically acceptable hydrates, solvates, or salts, have good stability due to the presence of disulfide bonds (compared to the monosulfide compounds in CN112691196). They are not easily oxidized by air, and their anticancer effects can be achieved through oral (oral) administration, rather than being limited to intraperitoneal or intravenous injection.

[0018] The inventors further discovered that the composition not only has extremely high anti-cancer activity, but also has good antibacterial effect. The combination of the two has a significant synergistic effect.

[0019] As a preferred embodiment of the present invention, at least one of the following (a) to (c) is provided:

[0020] (a) The R1 and R2 groups are selected from at least one of the following structures:

[0021]

[0022] (b) Choose from one of the following structures:

[0023]

[0024] (c) The structures of R5 and R6 are as follows:

[0025] As a preferred embodiment of the present invention, the compound represented by Formula I is panthioethylamine, furanthiamine, propoxur, alliin, ajoene, allyl trisulfide, dimethyl trisulfide, thiram, dihydroxyethyl disulfide, polycarpamine AE, pyrithione, PX-12, biotin disulfide, folic acid disulfide, or diphenyl disulfide.

[0026] The structural formula of the compound represented by Formula I is as follows:

[0027]

[0028]

[0029] As a preferred embodiment of the present invention, the compound represented by Formula II is thioctic acid, thioctic amide, romidixin, outovirin, varacins, leptosins, thiophene, thiomycin, aspergillin, or thiophene.

[0030] The structural formula of the compound represented by Formula II is as follows:

[0031]

[0032]

[0033] In a preferred embodiment of the present invention, the compound represented by Formula III is allicin.

[0034] The structural formula of the compound represented by Formula III is as follows:

[0035]

[0036] As a preferred embodiment of the present invention, the molar ratio of the polysulfide compound to aurinophene is ≥1, preferably ≥2, more preferably ≥200, and even more preferably ≥1000.

[0037] The inventors discovered that polysulfide compounds and aurnofen can only achieve a synergistic effect when combined in a specific molar ratio. In other words, polysulfide compounds and aurnofen only have a synergistic effect when combined in a specific molar ratio. When the amount of polysulfide compound added is too low, it has no synergistic effect on aurnofen.

[0038] When the molar ratio of polysulfide compound to aurnofen is 1:1, the polysulfide compound and aurnofen begin to exert a synergistic effect, and the synergistic effect is better as the molar ratio of polysulfide compound to aurnofen increases.

[0039] When the molar ratio of auronolactone to polysulfide compounds is 1:1 to 1000, the auronolactone and polysulfide compounds begin to exert a synergistic effect, and when the molar ratio is 1:1000, the anticancer activity and antibacterial activity reach their optimal levels.

[0040] In a preferred embodiment of the present invention, the polysulfide compound is panthionylamine, and the molar ratio of panthionylamine to aurinophene is ≥1, preferably ≥2, more preferably ≥200, and even more preferably ≥1000.

[0041] In a preferred embodiment of the present invention, the polysulfide compound is lipoic acid, and the molar ratio of lipoic acid to aurinophene is ≥1, preferably ≥2, more preferably ≥200, and even more preferably ≥1000.

[0042] In a preferred embodiment of the present invention, the polysulfide compound is thioctinamide, and the molar ratio of thioctinamide to aurinophene is ≥1, preferably ≥2, more preferably ≥200, and even more preferably ≥1000.

[0043] In a preferred embodiment of the present invention, the polysulfide compound is furfural thiamine, and the molar ratio of furfural thiamine to aurinophene is ≥1, preferably ≥2, more preferably ≥200, and even more preferably ≥1000.

[0044] In a preferred embodiment of the present invention, the polysulfide compound is propoxuridine, and the molar ratio of propoxuridine to aurinophene is ≥1, preferably ≥2, more preferably ≥200, and even more preferably ≥1000.

[0045] In a preferred embodiment of the present invention, the polysulfide compound is thiamine, and the molar ratio of thiamine to aurinophene is ≥1, preferably ≥2, more preferably ≥200, and even more preferably ≥1000.

[0046] In a preferred embodiment of the present invention, the polysulfide compound is ajoene, and the molar ratio of ajoene to aurinophene is ≥1, preferably ≥2, more preferably ≥200, and even more preferably ≥1000.

[0047] In a preferred embodiment of the present invention, the polysulfide compound is allicin, and the molar ratio of allicin to aurinophene is ≥1, preferably ≥2, more preferably ≥200, and even more preferably ≥1000.

[0048] In a preferred embodiment of the present invention, the polysulfide compound is allyl trisulfide, and the molar ratio of allyl trisulfide to aurinophen is ≥1, preferably ≥2, more preferably ≥200, and even more preferably ≥1000.

[0049] In a preferred embodiment of the present invention, the polysulfide compound is dimethyl trisulfide, and the molar ratio of dimethyl trisulfide to aurinophene is ≥1, preferably ≥2, more preferably ≥200, and even more preferably ≥1000.

[0050] In a preferred embodiment of the present invention, the polysulfide is a dihydroxyethyl disulfide, and the molar ratio of the dihydroxyethyl disulfide to aurinophene is ≥1, preferably ≥2, more preferably ≥200, and even more preferably ≥1000.

[0051] In a preferred embodiment of the present invention, the polysulfide compound is romidesin, and the molar ratio of romidesin to aurinophene is ≥1, preferably ≥2, more preferably ≥200, and even more preferably ≥1000.

[0052] In a preferred embodiment of the present invention, the polysulfide compound is polycarpamine AE, and the molar ratio of polycarpamine AE to aurinol is ≥1, preferably ≥2, more preferably ≥200, and even more preferably ≥1000.

[0053] In a preferred embodiment of the present invention, the polysulfide compound is Outovirin, and the molar ratio of Outovirin to aurinophene is ≥1, preferably ≥2, more preferably ≥200, and even more preferably ≥1000.

[0054] In a preferred embodiment of the present invention, the polysulfide compound is Varacins, and the molar ratio of Varacins to aurinophene is ≥1, preferably ≥2, more preferably ≥200, and even more preferably ≥1000.

[0055] In a preferred embodiment of the present invention, the polysulfide compound is leptosins, and the molar ratio of leptosins to aurinol is ≥1, preferably ≥2, more preferably ≥200, and even more preferably ≥1000.

[0056] In a preferred embodiment of the present invention, the polysulfide compound is thioclase, and the molar ratio of thioclase to aurinophene is ≥1, preferably ≥2, more preferably ≥200, and even more preferably ≥1000.

[0057] In a preferred embodiment of the present invention, the polysulfide compound is a pomycin, and the molar ratio of the pomycin to aurinol is ≥1, preferably ≥2, more preferably ≥200, and even more preferably ≥1000.

[0058] In a preferred embodiment of the present invention, the polysulfide compound is aspergillus, and the molar ratio of aspergillus to aurinol is ≥1, preferably ≥2, more preferably ≥200, and even more preferably ≥1000.

[0059] In a preferred embodiment of the present invention, the polysulfide compound is auriculin, and the molar ratio of auriculin to aurinophene is ≥1, preferably ≥2, more preferably ≥200, and even more preferably ≥1000.

[0060] In a preferred embodiment of the present invention, the polysulfide compound is pyrithione, and the molar ratio of pyrithione to aurinophene is ≥1, preferably ≥2, more preferably ≥200, and even more preferably ≥1000.

[0061] In a second aspect, the present invention proposes the use of the compounds described above in the preparation of antitumor drugs.

[0062] In a preferred embodiment of the present invention, the tumor is colon cancer, liver cancer, lung cancer, stomach cancer, cervical cancer, breast cancer, prostate cancer, bladder cancer, melanoma, or pancreatic cancer.

[0063] In a third aspect, the present invention proposes the use of the compositions described above in the preparation of cell proliferation inhibitors.

[0064] In a preferred embodiment of the present invention, the cells are colon cancer cells, liver cancer cells, lung cancer cells, gastric cancer cells, cervical cancer cells, breast cancer cells, prostate cancer cells, bladder cancer cells, melanoma cells, or pancreatic cancer cells.

[0065] In a fourth aspect, the present invention proposes the use of the compositions described above in the preparation of antibacterial drugs.

[0066] In a preferred embodiment of the present invention, the antibacterial drug is an antibacterial drug against Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, Pseudomonas aeruginosa, Acinetobacter baumannii, or Klebsiella pneumoniae.

[0067] In a fifth aspect of the invention, the use of the previously described composition in the preparation of TrxR enzyme inhibitors is proposed.

[0068] In a sixth aspect of the invention, the invention proposes the use of the polysulfide compounds described above as potentiators of aurinophene in the preparation of TrxR enzyme inhibitors.

[0069] The polysulfide compounds include compounds with structural formulas such as those shown in Formula I, Formula II, Formula III, and Formula IV; or

[0070] Isomers, pharmaceutically acceptable hydrates, solvates, or salts of compounds represented by Formulas I, II, III, and IV;

[0071]

[0072]

[0073] In Equations I and II, n is an integer ≥ 1;

[0074] Among them, R1, R2, R3, R4, R5, and R6 are independently selected from alkyl, alkenyl, carbonyl, aryl, alkyl derivatives containing heteroatoms, alkenyl derivatives containing heteroatoms, carbonyl derivatives containing heteroatoms, or aryl derivatives containing heteroatoms.

[0075] The heteroatom is at least one of N, S, O, P, and Si.

[0076] As a preferred embodiment of the present invention, at least one of the following (a) to (c) is provided:

[0077] (a) The R1 and R2 groups are selected from at least one of the following structures:

[0078]

[0079] (b) Choose from one of the following structures:

[0080]

[0081] (c) The structures of R5 and R6 are as follows:

[0082] As a preferred embodiment of the present invention, the compound represented by Formula I is panthioethylamine, furanthiamine, propoxur, alliin, ajoene, allyl trisulfide, dimethyl trisulfide, thiram, dihydroxyethyl disulfide, polycarpamine AE, pyrithione, PX-12, biotin disulfide, folic acid disulfide, or diphenyl disulfide.

[0083] The structural formula of the compound represented by Formula I is as follows:

[0084]

[0085]

[0086] As a preferred embodiment of the present invention, the compound represented by Formula II is thioctic acid, thioctic amide, romidixin, outovirin, varacins, leptosins, thiophene, thiomycin, aspergillin, or thiophene.

[0087] The structural formula of the compound represented by Formula II is as follows:

[0088]

[0089] In a preferred embodiment of the present invention, the compound represented by Formula III is allicin.

[0090] The structural formula of the compound represented by Formula III is as follows:

[0091]

[0092] As a preferred embodiment of the present invention, the molar ratio of the polysulfide compound to aurinophene is ≥1, preferably ≥2, more preferably ≥200, and even more preferably ≥1000.

[0093] In a seventh aspect of the invention, the invention provides a product comprising the composition described above, wherein the product is an antitumor drug, an antibacterial drug, or a cell proliferation inhibitor.

[0094] In a preferred embodiment of the present invention, the product further includes pharmaceutically acceptable excipients. Those skilled in the art can conventionally select suitable excipients based on the dosage form and overall properties of the drug.

[0095] As a preferred embodiment of the present invention, at least one of the following (a) to (c) is provided:

[0096] (a) The tumor is colon cancer, liver cancer, lung cancer, stomach cancer, cervical cancer, breast cancer, prostate cancer, bladder cancer, melanoma, or pancreatic cancer;

[0097] (b) The cells are colon cancer cells, liver cancer cells, lung cancer cells, gastric cancer cells, cervical cancer cells, breast cancer cells, prostate cancer cells, bladder cancer cells, melanoma cells, or pancreatic cancer cells;

[0098] (c) The antibacterial drug is an antibacterial drug against Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, Pseudomonas aeruginosa, Acinetobacter baumannii, or Klebsiella pneumoniae.

[0099] The beneficial effects of this invention are as follows:

[0100] (1) Under normal physiological conditions, aurinophene cannot effectively inhibit the proliferation of human colon cancer cells, human liver cancer cells, human lung cancer cells, human gastric cancer cells, human cervical cancer cells, human breast cancer cells, human prostate cancer cells, human bladder cancer cells, human melanoma cells, and pancreatic cancer. However, when non-toxic doses of polysulfide compounds (such as panthioethylamine, lipoic acid, and furazolidone) are added, aurinophene exhibits excellent inhibitory effects on all 10 types of cancer cells, including inhibition of TrxR and tumor cell proliferation. In vivo tumor model experiments in mice also show that aurinophene can effectively inhibit tumor growth when combined with polysulfide compounds, indicating that the combination of aurinophene and polysulfide compounds can be used as an anticancer drug combination. Furthermore, since the types of polysulfide compounds described in this invention can be adjusted, the absorption of aurinophene in tumor tissue can be regulated, thereby exhibiting good selectivity in in vivo anticancer activity.

[0101] (2) The composition has good stability, is not easily oxidized by air, and can achieve anticancer effects through oral administration (gavage administration), rather than being limited to intraperitoneal or intravenous injection.

[0102] (3) When the composition is used in combination with aurinophene and polysulfide compounds, it has good antibacterial effect. Attached Figure Description

[0103] Figure 1 This figure shows the effect of aurinol combined with panthioethylamine on tumor volume in a mouse tumor model.

[0104] Figure 2 The figure shows the effect of aurinol combined with panthioethylamine on mouse body weight in a mouse tumor model.

[0105] Figure 3 The figure shows the effect of aurinol combined with panthioethylamine on tumor quality in a mouse tumor model.

[0106] Figure 4 This image shows the actual tumor-suppressive effect of aurinol combined with panthioethylamine in a mouse tumor model (tumor image). Detailed Implementation

[0107] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0108] Unless otherwise specified, all reagents or instruments used in this invention are commercially available products.

[0109] Example

[0110] The compositions of Examples 1 to 29 are shown in Table 1, where the molar ratios represent the corresponding polysulfide compounds to aurinophene molar ratios.

[0111] Table 1

[0112]

[0113]

[0114]

[0115] Example 30

[0116] 1. Effects of aurinophene combined with various polysulfide compounds on the inhibitory activity of multiple cancer cells.

[0117] (1) Culture of cancer cells

[0118] Take the culture flask containing the desired cancer cells, discard the original culture medium, wash three times with 2 mL PBS, add 1 mL trypsin for 30 seconds, and add 2 mL culture medium to stop the digestion. Transfer the cells to a centrifuge tube, centrifuge at 1000 rpm for 3 minutes, discard the supernatant, add 2 mL culture medium, and mix by pipetting 10 times. Take 10 μL of the cell suspension into a cell counter and count the cells; seed 5000 cells per well, with 3 replicates per compound, and calculate the required cell volume and well volume. Calculate the required cell suspension volume using cell counting, dilute the required number of cells with culture medium to 5000 cells per well, and seed with 100 μL of culture medium. Mix by pipetting, seed the cells using a pipette, and incubate in an incubator, labeling as desired.

[0119] (2) Take the cells cultured in step (1), discard the original culture medium, dilute the aurinophen stock solution to a certain concentration, mix well, use a pipette to draw 100 μL of diluent per well, tilt the 96-well plate, add the aurinophen diluent to the 96-well plate (to prevent cell damage), mark it, gently shake it left and right and back and forth 3 times, and put it into a CO2 incubator for culture.

[0120] After adding aurinophen and culturing for 1 hour, dilute the stock solution containing polysulfide compounds to a certain concentration, mix well, and use a pipette (using 3 wells) to aspirate 20 μL of diluent to each well. Tilt the 96-well plate and add the diluent to the corresponding 3 wells (to prevent cell damage). Add different polysulfide compounds in sequence, and mix well by pipetting for about 3 minutes after each addition. Return the plate to the incubator for further culture.

[0121] (3) MTT assay for detecting the inhibitory effect on cancer cell proliferation

[0122] After treating cells with different polysulfide compounds for 24 hours, 20 μL of MTT (5 mg / mL in PBS) was added to each well of a 96-well plate using a multipipeline. The cells were incubated at 37°C for 4 hours. After incubation, the MTT mixture was removed, and DMSO was added at 130 μL / well using a multipipeline. The cells were shaken horizontally for 10 minutes until the formazan was fully dissolved. The absorbance was measured at 490 nm using a microplate reader. Cell viability at each drug concentration was calculated as required, and a scatter plot was plotted. The effects of aurinol, polysulfide compounds, and the combination of the two drugs on cancer cell proliferation are shown in Tables 2 and 3.

[0123] Note (Table 2): For the following four cell lines, namely lung cancer cells (PCGR, A549), cervical cancer cells (HeLa), and breast cancer cells (MCF-7), the dosage of aurinophene is 10 μM; for the following other cell lines, the dosage of aurinophene is 5 μM.

[0124] Table 2. Effects of the combined use of aurinophene and panthiophene (200 μM) on cell viability in different cell lines.

[0125]

[0126]

[0127] Table 3. Effects of auronoxine (5 μM) and various polysulfide compounds (50 μM), and their combination, on the cell survival rate of the lung cancer cell line (PC9) (in this experiment, polysulfide compounds were used at non-toxic or low-toxic concentrations).

[0128]

[0129]

[0130] As shown in Table 2-3, under normal physiological conditions (the addition of FBS to the empty culture medium can simulate the high-sulfhydryl environment under physiological conditions), aurinofen showed no significant inhibitory effect on human colon cancer cells, human liver cancer cells, human lung cancer cells, human gastric cancer cells, human cervical cancer cells, human breast cancer cells, human prostate cancer cells, human bladder cancer cells, human melanoma cells, and pancreatic cancer. However, when non-toxic doses of polysulfide-containing compounds (such as panthioethylamine, thioctic acid, thioctic amide, furazolidone, propoxythiamine, allithiamine, ajoene, allicin, allyl trisulfide, dimethyl trisulfide, thiram, 2-Hydroxyethyldisulfide, romidepsin, and polycarpamine) were added, the inhibitory effect was significantly reduced. AE, Outovirin, Varacins, Leptosins, Thiolutin, Holomycin, Gliotoxin, Aureothricin, Pyrithioxine, and Aurenofen all showed excellent inhibitory effects on the above 10 types of cancer cells.

[0131] 2. Study on the ratio range of aurinophene and polysulfide compounds used in combination

[0132] Table 4. Effects of auronoxine and different molar concentrations of pan-thioethylamine, and their combination, on the cell survival rate of the lung cancer cell line (PC9).

[0133]

[0134]

[0135] Note: The ratio is [Aurinophene concentration: Panthiophene concentration]; here the aurinophene concentration is fixed at 5μM, and the panthiophene concentration is changed.

[0136] The results in Table 4 show that when the concentration (ratio) of the polysulfide compound is low, it has no synergistic effect with aurnofen. The synergistic effect only occurs when the concentration (ratio) of the polysulfide compound increases. Therefore, we propose that a synergistic effect is only achieved when the ratio of polysulfide compound to aurnofen is greater than or equal to 1, and the synergistic effect increases with the amount of polysulfide compound used.

[0137] 3. Enhancement of the inhibitory effect of polysulfide compounds on the TrxR enzyme activity of aurinophene

[0138] The testing steps are as follows:

[0139] A549 cells were used as the experimental subject.

[0140] Cells were loaded at 2×10 5 Inoculate one cell per well in a 6-well plate and incubate for 24 hours.

[0141] After incubation, the cell culture medium was replaced with 30% FBS to simulate a high-concentration thiol environment under normal physiological conditions. A control group was maintained without any reagents, while the experimental group was incubated with 1.0 μM aurinophene at 37°C for 30 min. Polysulfide compounds (panthioethylamine, furazolidone) were added and incubated for 12 h (final DMSO concentration ≤1%). After incubation, the cells were washed three times with PBS, and 100 μL of ice-cold lysis buffer (containing 50 mM pH 7.4 phosphate buffer, 1 mM EDTA, and 0.1% Triton-X 100) was added. Cell lysis was performed on ice for 5 min, and the cell lysate was collected.

[0142] 100 μL of buffer (containing 50 mM potassium phosphate at pH 7.4, 1 mM EDTA, and 0.2 mM NADPH) was added to the collected cell lysate (10 μg protein). 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB, final concentration 3 mM) was then added to initiate the reaction. TrxR activity was determined by the increase in OD value over 10 minutes (OD value detection wavelength was 410 nm). Aurinophene alone served as a positive control, and the blank control served as a negative control. The results are shown in Table 5.

[0143] Table 5. Inhibitory effects of each experimental group on TrxR enzyme activity

[0144]

[0145]

[0146] Note (Table 5): * indicates that the culture medium contains 30% FBS.

[0147] Table 5 shows that under FBS-free conditions (i.e., simulated in vitro conditions), aurinophene can inhibit TrxR enzyme activity in A549 cells to 13.2%; however, under 30% FBS conditions (simulated in vivo conditions), the ability of aurinophene to inhibit enzyme activity decreases, only inhibiting intracellular TrxR enzyme activity to 82.1%. However, when polysulfide compounds are added, even under 30% FBS conditions, the ability of aurinophene to inhibit enzyme activity is greatly improved; compared to the absence of polysulfide compounds, the ability of aurinophene to inhibit enzyme activity can be increased by more than three times.

[0148] 4. Effects of aurinophen combined with various polysulfide compounds on aurinophen cellular uptake.

[0149] The testing steps are as follows:

[0150] A549 cells were used as the experimental subject.

[0151] Cells were loaded at 2×10 5 Inoculate one cell per well in a 6-well plate and incubate for 24 hours.

[0152] After incubation, the cell culture medium was replaced with 30% FBS to simulate a high-concentration thiol environment under normal physiological conditions. A control group was maintained without any reagents, while the experimental group was incubated with 2 μM aurinolone at 37°C for 10 min. A polysulfide compound (50 μM furfural) was then added for incubation for 12 h (the final concentration of DMSO in the system was ≤1%). Immediately after incubation, the cells were washed three times with PBS, and 500 μL of ultrapure water was added to each well to lyse the cells. After 15 min, the cell lysate was collected.

[0153] The collected cell lysates were dissolved in aqua regia and then diluted with ultrapure water to an appropriate ratio. Gold was detected using inductively coupled plasma mass spectrometry (ICP-MS). The cell uptake rate of 5 μM auronoxine without FBS was used as a control. The results are shown in Table 6.

[0154] Table 6. Effects of furazolidone on cellular uptake of aurinophene

[0155]

[0156]

[0157] The above-mentioned absorption amount refers to the gold content (mg) per gram of protein in the cell.

[0158] Table 6 shows that, under FBS-free conditions, with auronoxine as a reference, the gold absorption rate reached 100%. However, when the concentration of FBS in the culture medium was increased, the gold absorption decreased rapidly; when furazolidone was added, the absorption of auronoxine increased significantly. This indicates that the addition of furazolidone improved the intracellular absorption of auronoxine.

[0159] 5. Actual inhibitory effect of aurinophene combined with polysulfide compounds in a mouse tumor model.

[0160] The testing steps are as follows:

[0161] Establishing a mouse tumor model: Two million HCT116 colon cancer cells suspended in PBS were subcutaneously injected into the dorsal side of 5-7 week old female BALB / c-nu / nu (nude mice) to establish a xenograft model. When the tumor volume reached approximately 50 mm², the xenograft model was established. 3(3-4 days after tumor inoculation) Mice were randomly divided into a control group (using castor oil) and a treatment group (auronoxine group, panthiophene group, and combined treatment group). The combined treatment group was treated with auronoxine (10 mg / kg mouse body weight / day) combined with panthiophene (200 mg / kg mouse body weight / day) by gavage, twice a week. When the tumor volume reached 1000 mm², the treatment was continued. 3 Afterwards, the mice were anesthetized and then euthanized by dislocating their cervical vertebrae.

[0162] The results are as follows Figure 1-4 As shown.

[0163] Tumor growth was not effectively inhibited in the control group, the aurinophen monotherapy group, or the polysulfide monotherapy group; however, tumor growth was significantly inhibited when aurinophen was used in combination with polysulfide.

[0164] 6. Effects of aurinol in combination with various polysulfide compounds on the inhibitory activity of multiple bacteria.

[0165] The testing steps are as follows:

[0166] (1) Bacterial transfer

[0167] Use an inoculation loop to pick out a single bacterial clone from LA solid medium, then transfer it to a 50ml centrifuge tube containing 2ml of LB medium, and incubate it overnight at 37°C in a shaker. Once the medium becomes turbid, it can be used for experiments.

[0168] (2) Take a small amount of the bacterial solution from step 1 into 2 mL of culture medium (broth medium), place it in a 50 mL centrifuge tube, and incubate it at an angle in a shaker at 37 °C.

[0169] (3) After 12 hours, dilute the bacterial solution (20uL) from step 2 to the culture medium (2ml), mix them in a 50ml centrifuge tube, and place it at an angle in a shaker at 37℃ for incubation.

[0170] (4) Adding medicine

[0171] The corresponding drug concentrations were added to the 96-well plate in advance to make the final concentration of aurinophen 40 μM and the concentration of polysulfide compounds fixed at 200 μM.

[0172] After 2.5 hours, take out the bacterial solution from step 3, dilute it 1000 times in the dosing tank, and then add the diluted bacterial solution to the 96-well plate at 150 μL / well. Place the plate in a shaker at 37°C to fix it.

[0173] After 24 hours, the absorbance was measured at a wavelength of 600 nm using an ELISA reader. The bacterial inhibition rate at each drug concentration was calculated as required, and a scatter plot was plotted. The results of the effects of aurinol, polysulfide compounds, and the combination of the two drugs on the growth of the bacterial strain are shown in Table 7.

[0174] Staphylococcus aureus (ATCC29213); Staphylococcus epidermidis (ATCC12228); Escherichia coli (ATCC25922); Pseudomonas aeruginosa (ATCC9027); Acinetobacter baumannii (ATCC19606); Klebsiella pneumoniae (ATCC10031).

[0175] Table 7. Effects of auronorfen (40 μM) and panthiophene (200 μM), and their combination, on the inhibitory activities of various bacteria.

[0176]

[0177] Antibacterial rate = (Number of colonies in control group - Number of colonies in treatment group) / Number of colonies in control group * 100%

[0178] As shown in Table 7, under normal physiological conditions (the addition of FBS to the empty culture medium can simulate the high thiol environment under physiological conditions), aurinofen showed no significant inhibitory effect on Staphylococcus aureus (ATCC29213); Staphylococcus epidermidis (ATCC12228); Escherichia coli (ATCC25922); Pseudomonas aeruginosa (ATCC9027); Acinetobacter baumannii (ATCC19606); and Klebsiella pneumoniae (ATCC10031). However, when a non-toxic dose of aurinofen containing a polysulfide compound (panthionylamine) was added, it showed excellent inhibitory effects on all six strains.

[0179] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit the scope of protection of the present invention. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the essence and scope of the technical solutions of the present invention.

Claims

1. A composition characterized in that, The composition comprises aurothioglucose and a polysulfide; the polysulfide comprises at least one of pantethine, fursultiamine, propylthiuracil, allitridin, ajoene, allyl trisulfide, dimethyl trisulfide, thiram, dihydroxyethyl disulfide, thioctic acid, thioctic amide, allicin; The molar ratio of the polysulfide to aurothioglucose is ≥10.

2. The composition of claim 1, wherein, The molar ratio of the polysulfide to aurothioglucose is ≥200.

3. The composition of claim 1, wherein, The molar ratio of the polysulfide to aurothioglucose is ≥1000.

4. The composition of claim 1, wherein, The polysulfide is pantethine, and the molar ratio of the pantethine to aurothioglucose is ≥10; or The polysulfide is thioctic acid, and the molar ratio of the thioctic acid to aurothioglucose is ≥10; or The polysulfide is thioctic amide, and the molar ratio of the thioctic amide to aurothioglucose is ≥10; or The polysulfide is fursultiamine, and the molar ratio of the fursultiamine to aurothioglucose is ≥10; or The polysulfide is propylthiuracil, and the molar ratio of the propylthiuracil to aurothioglucose is ≥10; or The polysulfide is allitridin, and the molar ratio of the allitridin to aurothioglucose is ≥10; or The polysulfide is ajoene, and the molar ratio of the ajoene to aurothioglucose is ≥10; or The polysulfide is allicin, and the molar ratio of the allicin to aurothioglucose is ≥10; or The polysulfide is allyl trisulfide, and the molar ratio of the allyl trisulfide to aurothioglucose is ≥10; or The polysulfide is dimethyl trisulfide, and the molar ratio of the dimethyl trisulfide to aurothioglucose is ≥10; or The polysulfide is dihydroxyethyl disulfide, and the molar ratio of the dihydroxyethyl disulfide to aurothioglucose is ≥10.

5. Use of the composition according to any one of claims 1 to 4 for the manufacture of an antitumor medicament, characterized in that, The tumor is colon cancer, liver cancer, non-small cell lung cancer, gastric cancer, cervical cancer, breast cancer, prostate cancer, bladder cancer, melanoma or pancreatic cancer.

6. Use of the composition according to any one of claims 1 to 4 for the manufacture of a cell proliferation inhibitor, characterized in that, The cell is a colon cancer cell, a liver cancer cell, a non-small cell lung cancer cell, a gastric cancer cell, a cervical cancer cell, a breast cancer cell, a prostate cancer cell, a bladder cancer cell, a melanoma cell or a pancreatic cancer cell.

7. Use of a composition for the manufacture of an antibacterial medicament, characterized in that, The composition comprises aurothioglucose and pantethine, and the molar ratio of the pantethine to aurothioglucose is ≥5. The antibacterial drug is an anti-Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, Pseudomonas aeruginosa, Acinetobacter baumannii or Klebsiella pneumoniae drug.

8. Use according to claim 7, characterized in that, The molar ratio of the pantethine to aurothioglucose is ≥200.

9. Use according to claim 7, characterized in that, The molar ratio of the pantethine to aurothioglucose is ≥1000.

10. A product characterized by, The product is an antitumor drug or a cell proliferation inhibitor; the tumor is colon cancer, liver cancer, non-small cell lung cancer, gastric cancer, cervical cancer, breast cancer, prostate cancer, bladder cancer, melanoma or pancreatic cancer; the cell is a colon cancer cell, a liver cancer cell, a non-small cell lung cancer cell, a gastric cancer cell, a cervical cancer cell, a breast cancer cell, a prostate cancer cell, a bladder cancer cell, a melanoma cell or a pancreatic cancer cell.

11. The product of claim 10, wherein, It also comprises a pharmaceutically acceptable adjuvant.

12. A product characterized by, A composition comprising aurofim and pentetrazol, the product being an antibacterial agent, the molar ratio of pentetrazol to aurofim being > 5; the antibacterial agent being an agent against Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, Pseudomonas aeruginosa, Acinetobacter baumannii or Klebsiella pneumoniae.

13. The product of claim 12, wherein, The molar ratio of pentetrazol to aurofim is > 200.

14. The product of claim 12, wherein, The molar ratio of pentetrazol to aurofim is > 1000.

Images