Ruthenate pharmaceutical composition as well as preparation method and application thereof

Abstract

The invention discloses a ruthenium acid salt pharmaceutical composition and a preparation method and application thereof, the ruthenium acid salt pharmaceutical composition comprises trans-[tetrachloro bis (1H-indazole) ruthenium (III) acid salt] and a medical nitrogen-containing heterocyclic compound, and the medical nitrogen-containing heterocyclic compound comprises metiradone and / or acetazolamide. The medical nitrogen-containing heterocyclic compound of metiradone and / or acetazolamide disclosed by the invention is combined with a trans-[tetrachloro bis (1H-indazole) ruthenium (III) acid salt] compound for use, so that tumor growth can be well inhibited, and the medical nitrogen-containing heterocyclic compound is high in safety, free of pungent smell and good in chemical stability.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of biological medicine, in particular to a ruthenate pharmaceutical composition and a preparation method and application thereof. BACKGROUND

[0002] BOLD-100 (CAS No. 197723-00-5) is a kind of intravenous injection of ruthenium-based antitumor clinical candidate drug, which has passed a clinical trial (clinical trial number: NCT01415297). In the completed phase II clinical experiment, BOLD-100 is combined with 5-fluorouracil, calcium folinate and oxaliplatin to treat solid tumors such as colorectal cancer, pancreatic cancer and cholangiocarcinoma. However, studies have shown that BOLD-100 is prone to hydrolysis under physiological conditions, and irreversible binding with proteins occurs, resulting in the destruction of its own structure, difficulty in entering cells, and reduced antitumor activity. As a result, the effect of BOLD-100 on inhibiting tumor growth in several solid tumor models (colon cancer, breast cancer) is general. In recent years, there have been a few studies on the anticancer activity of BOLD-100 combined with other compounds, but the effect is not obvious. SUMMARY

[0003] The present application aims to at least solve one of the above technical problems existing in the prior art. To this end, the purpose of the present application is to provide a ruthenate pharmaceutical composition and a preparation method and application thereof.

[0004] In order to achieve the above-mentioned purpose, the technical solution adopted by the present application is:

[0005] The first aspect of the present application provides a ruthenate composition, comprising trans-[tetrachlorobis(H-indazole)ruthenate(III)] and a pharmaceutically acceptable nitrogen-containing heterocyclic compound, wherein the pharmaceutically acceptable nitrogen-containing heterocyclic compound comprises metyrapone and / or acetazolamide. 1 H-indazole) ruthenate(III)] and a pharmaceutically acceptable nitrogen-containing heterocyclic compound, wherein the pharmaceutically acceptable nitrogen-containing heterocyclic compound comprises metyrapone and / or acetazolamide.

[0006] Although the inventors' previous studies have shown that the combination of 2-methylthiophenylboronic acid and BOLD-100 can also exhibit a strong tumor inhibitory effect, the sulfide group in the 2-methylthiophenylboronic acid structure produces a thiol odor similar to garlic or rotten eggs. The boronic acid group has a sour taste, giving it a strong pungent odor. As a medicinal substance, it can easily cause nausea / vomiting, reducing medication compliance. Long-term inhalation also poses a potential risk of respiratory irritation. Furthermore, the boronic acid bond (BO) and sulfide bond (CS) of 2-methylthiophenylboronic acid are extremely susceptible to cleavage or oxidation, and the resulting byproducts are prone to causing unknown toxic side effects. The unstable nature of 2-methylthiophenylboronic acid also requires it to be stored under harsh conditions. The boronic acid group hydrolyzes after absorbing moisture, and it needs to be stored dry at -20°C to prevent hydrolysis, which increases storage costs. 2-methylthiophenylboronic acid also contains a hydrophobic benzene ring and a polar boronic acid group. At physiological pH, the boronic acid group forms an anion [B(OH)3] - However, the hydrophobic region within the molecule hinders dissolution and is difficult to improve through conventional solubilization techniques such as cyclodextrin inclusion. Because it easily reacts with the hydroxyl group of cyclodextrin, its oral bioavailability is close to 0. As mentioned above, it decomposes rapidly in water and cannot be stably present in injections. 2-methylthiophenylboronic acid also has potential toxic metabolite accumulation, such as the hydrolysis product phenylboronic acid, which can cause kidney damage. In addition, studies on the chelate formed when 2-methylthiophenylboronic acid is used in combination with BOLD-100 also found that, Figures 1 to 3 Under high-resolution mass spectrometry, the two formed different forms of chelates (the structural formula of chelate 1 is: The structural formula of chelate 2 is: The structural formula of chelate 3 is: ), resulting in uncontrollable biological behavior. Different chelate ring structures (five-membered and six-membered) vary in stability, reactivity, pharmacokinetics, and biological targeting, which is the molecular basis of potential toxicity. These aspects make it difficult to use as a clinical drug.

[0007] In the present invention, the medicinal nitrogen-containing heterocyclic compound and trans-[tetrachlorobis( 1 H-indazole) ruthenium (III) salt] combined with [not only shows a strong tumor inhibitory effect, but metyrapone and acetazolamide are both clinical drugs and have been used clinically for more than 50 years. They have a clear mechanism of action, high safety, high oral bioavailability, and metyrapone can also be used to treat patients through intravenous injection. It does not have a pungent odor, has high medication compliance, has no special requirements for storage conditions, is low in moisture sensitivity, and has high chemical stability. It solves the defects of the aforementioned 2-methylthiophenylboronic acid and also solves the problem of trans-[tetrachlorobis( 1 H-indazole)ruthenate(III) salts] have the disadvantage of poor pharmacological activity under physiological conditions.

[0008] In some embodiments of the present invention, the medicinal nitrogen-containing heterocyclic compound and trans-[tetrachlorobis( 1 H-indazole)ruthenate (III) salt] is in a molar ratio of (0.1-50):1, such as (1-45):1, (1-30):1, (1-25):1, (1-20):1, (1-18):1, (1-10):1, (1-8):1, etc.

[0009] In some embodiments of the present invention, the metyrapone and trans-[tetrachlorobis( 1 H-indazole)ruthenate (III) salt] is in a molar ratio of (0.1-50):1, such as (1-45):1, (1-30):1, (1-25):1, (1-20):1, (1-18):1, (1-10):1, (1-8):1, etc.

[0010] In some embodiments of the present invention, the acetazolamide and trans-[tetrachlorobis( 1 H-indazole)ruthenate (III) salt] is in a molar ratio of (0.1-50):1, such as (1-45):1, (1-30):1, (1-25):1, (1-20):1, (1-18):1, (1-10):1, (1-8):1, etc.

[0011] In some embodiments of the present invention, the trans-[tetrachlorobis( 1 The structural formula of [H-indazole) ruthenate (III) salt] is shown in Formula I: Wherein, R is selected from Na + 、 trans-[tetrachlorobis( 1 H-indazole)ruthenate (III) salt] will produce precipitation over time in PBS solution, and the solution will turn blue under full incubation conditions. 1 After entering the human body, [H-indazole] ruthenium(III) salt will quickly non-covalently bind to human serum albumin (HSA), but after a period of time, irreversible covalent binding will occur, which will reduce the cellular uptake of the ruthenium complex and prevent it from exerting an effective tumor inhibitory effect.

[0012] In some embodiments of the present invention, the trans-[tetrachlorobis( 1 H-indazole)ruthenate(III)] includes and / or

[0013] In some embodiments of the present invention, the ruthenate composition further comprises a pharmaceutically acceptable excipient.

[0014] The composition of the present application can be administered to a patient or subject in need of such treatment by any route, including oral, parenteral, rectal, or pulmonary administration. When administered orally, the pharmaceutical composition can be formulated into oral preparations, such as conventional oral solid preparations, e.g., tablets, capsules, pills, granules, etc., or oral liquid preparations, e.g., oral solutions, oral suspensions, syrups, etc. When formulated into oral preparations, suitable fillers, binders, disintegrants, lubricants, etc. can be added. When administered parenterally, the pharmaceutical composition can be formulated into injections, including injection solutions, sterile powders for injections, and concentrated solutions for injections. When formulated into injections, they can be produced by conventional methods in the pharmaceutical field, and can be prepared without adding additional agents or by adding suitable additional agents depending on the nature of the drug. When administered rectally, the pharmaceutical composition can be formulated into suppositories, etc. When administered by the pulmonary route, the pharmaceutical composition can be formulated into inhalation preparations, aerosol preparations, powder aerosol preparations, or spray preparations, etc.

[0015] The pharmaceutically acceptable excipient refers to a substance that is nontoxic, compatible with the active ingredient, and otherwise biologically suitable for use in the body. The choice of a particular excipient will depend on the mode of administration for treating a particular patient or the type and state of the disease. Examples of the pharmaceutically acceptable excipient include, but are not limited to, solvents, diluents, dispersants, suspending agents, surfactants, isotonic agents, thickening agents, emulsifiers, binders, lubricants, stabilizers, hydrating agents, emulsification accelerators, buffers, absorbents, coloring agents, ion exchangers, release agents, coating agents, flavoring agents, and antioxidants, etc. If necessary, flavoring agents, preservatives, and sweetening agents, etc. can also be added to the pharmaceutical composition.

[0016] Tablets can contain inert binding agents, for example, calcium carbonate, calcium phosphate, sodium carbonate or lactose; granulating and disintegrating agents, for example, corn starch or alginic acid; binding agents, for example, starch, gelatin or acacia; and lubricating agents, for example, aluminum or magnesium stearate, talc or silica. They can also have coatings, which are prepared such as to cause a delayed release of the pharmaceutical preparation in the gastrointestinal tract and reabsorption, for example, to achieve improved compatibility, assimilation or retardation. Gelatin capsules can contain the pharmaceutical substance with a solid, for example, calcium carbonate or kaolin, or an oily substance, for example, olive oil, peanut oil or paraffin oil diluent.

[0017] Aqueous suspensions may contain suspending agents, for example, sodium carboxymethylcellulose, methylcellulose, hydroxypropylcellulose, sodium alginate, polyvinylpyrrolidone, gum tragacanth or gum arabic; dispersants or wetting agents, for example, polyoxyethylene stearate, heptadecanol, polyoxyethylene sorbitan monooleate or lecithin; preservatives, for example, methyl hydroxybenzoate or propyl hydroxybenzoate; flavor regulators; sweeteners, for example, sucrose, lactose, sodium cyclamate, glucose, invert sugar syrup.

[0018] Oily suspensions may be, for example, peanut oil, olive oil, sesame oil, coconut oil or paraffin oil and a thickener, such as beeswax, a high melting point wax or cetyl alcohol; as well as sweeteners, taste modifiers and antioxidants.

[0019] Water-dispersible powders and granules may contain a composition of the invention in admixture with dispersing, wetting and suspending agents, such as those mentioned above, and sweetening, flavoring and coloring agents.

[0020] Emulsions may contain, for example, olive oil, peanut oil or paraffin oil together with emulsifiers such as arabinose, tragacanth, phospholipids, sorbitan monooleate, polyoxyethylene sorbitan monooleate and sweeteners and flavorings.

[0021] Aqueous solutions may contain preservatives such as methyl or propyl hydroxybenzoate; thickeners; flavor enhancers; sweeteners such as sucrose, lactose, cyclamate, glucose, invert sugar syrup; and flavoring and coloring agents.

[0022] Lyophilized preparations for injection generally include the composition of the present invention, a pH buffer and a cryoprotectant. The general method for providing the preparation comprises the following steps: preparing an aqueous buffer solution, preparing an aqueous cryoprotectant solution, dissolving the composition of the ruthenate in a buffer solution, adding the cryoprotectant solution, sterilizing and filtering (e.g., aseptic filtration), filling a vial under aseptic conditions, and lyophilizing under aseptic conditions. Suitable buffers include, but are not limited to, citrate, TRIS, acetate, EDTA, HEPES, tricine (N-tris (hydroxymethyl) methylglycine) and imidazole. Using phosphate buffer is possible, but not preferred. A preferred aspect of the present invention is to use citric acid / sodium citrate buffer. Suitable cryoprotectants include, but are not limited to, sugars, monosaccharides, disaccharides, polyols, mannitol, sorbitol, sucrose, trehalose, dextran and dextrose.

[0023] The second aspect of the present invention provides a use of the ruthenate composition in the preparation of anti-tumor drugs.

[0024] The term "tumor" should be understood to refer to all forms of neoplastic cell growth, including tumors of the lung, liver, skin, pancreas and colon.

[0025] In some embodiments of the present invention, the tumor comprises at least one of colorectal cancer, liver cancer, lung cancer, brain cancer, cervical cancer, breast cancer, melanoma, pancreatic cancer, multiple myeloma (MM), or blood cancer.

[0026] In some embodiments of the present invention, the lung cancer includes small cell lung cancer, non-small cell lung cancer, lung adenocarcinoma and lung squamous cell carcinoma.

[0027] In some embodiments of the present invention, the brain cancer includes glioma and non-glioma. The most common brain tumor is glioma, including astrocytoma, oligodendroglioma and ependymoma; and non-glioma includes embryonal tumor, meningioma, craniopharyngioma, schwannoma, ganglioglioma, pituitary adenoma, and choroid plexus tumor.

[0028] In some embodiments of the present invention, the therapeutically effective amount of the ruthenate composition is calculated based on the therapeutically effective amount of trans-[tetrachlorobis(1H-indazole)ruthenate(III) salt]. For example, the therapeutically effective amount of trans-[tetrachlorobis(1H-indazole)ruthenate(III) salt] is 320 mg / m 2 Up to 625 mg / m 2 The amount of body surface area (BSA) of the patient can be calculated using Modified Dubois, that is, BSA (m 2 )=0.007184xHeight(cm) 0.725 x weight (kg) 0.425 .

[0029] In some embodiments of the present invention, based on the combined therapeutic effects of the components in the ruthenate composition of the present invention, the therapeutically effective amount of trans-[tetrachlorobis(1H-indazole)ruthenate(III) salt] is reduced to 0.05 to 0.95 times, such as 0.09 to 0.9 times, 0.09 to 0.8 times, or 0.09 to 0.7 times, of the dosage of the single agent.

[0030] A "therapeutically effective amount" refers to an amount effective to achieve the desired therapeutic outcome, at the dosage and for the duration necessary. The therapeutically effective amount of a formulation can vary depending on factors such as the disease state, the age, sex, and weight of the individual, and the ability of the compound to elicit the desired response in the individual. The dosage regimen can be adjusted to provide the optimal therapeutic response. A therapeutically effective amount can also be an amount in which the therapeutically beneficial effects outweigh any toxic or adverse effects of the formulation or active compound.

[0031] The beneficial effects of the present invention are:

[0032] The combination of the medicinal nitrogen-containing heterocyclic compound of metyrapone and / or acetazolamide and the trans-[tetrachlorobis(1H-indazole)ruthenate(III) salt] compound of the present invention can effectively inhibit tumor growth, has high safety, no irritating odor, and good chemical stability, and metyrapone can also be administered by injection.

[0033] The present invention addresses the shortcomings of trans-[tetrachlorobis(1H-indazole)ruthenate(III)] under normal physiological conditions, including its inherent structural instability, easy hydrolysis, and irreversible protein binding, which results in its inability to effectively inhibit cancer cell proliferation. Cell experiments demonstrate that the combined use of metyrapone and / or acetazolamide with the ruthenium complex is far more effective than the ruthenium complex alone.

[0034] The metyrapone and / or acetazolamide of the present invention can effectively inhibit the formation of multinuclear ruthenium aggregates and enhance the anticancer activity thereof. BRIEF DESCRIPTION OF THE DRAWINGS

[0035] Figures 1 to 3 High-resolution mass spectra of chelate 1, chelate 2, and chelate 3 formed by 2-methylthiophenylboronic acid and BOLD-100, respectively.

[0036] Figure 4 This figure shows the effect of BOLD-100 combined with metyrapone on tumor volume in a mouse tumor model.

[0037] Figure 5 This figure shows the effect of BOLD-100 combined with metyrapone on the body weight of mice in a mouse tumor model.

[0038] Figure 6 Figure 1 shows the actual tumor inhibition effect (tumor image) of the BOLD-100 combined with metyrapone in a mouse tumor model; each row is a group, and from top to bottom are the control group, metyrapone group, BOLD-100 group, and BOLD-100 combined with metyrapone group.

[0039] Figure 7These are photographs of fresh livers after dissection of mice in each drug-treated group of the present invention; each row is a group, and from top to bottom they are the control group, metyrapone group, BOLD-100 group, and BOLD-100 combined with metyrapone group.

[0040] Figure 8 This is the UV-visible absorption spectrum of BOLD-100 (400 μM) of the present invention in PBS from 0 to 480 min.

[0041] Figure 9 The UV-visible absorption spectra of BOLD-100 (200 μM) of the present invention and its mixture with metyrapone or acetazolamide in PBS (700 nm leveling). DETAILED DESCRIPTION

[0042] The present invention is further described in detail below using specific examples. Unless otherwise specified, the raw materials, reagents, and apparatus used in the examples and comparative examples are available from conventional commercial sources or by conventional methods. Unless otherwise specified, all experimental or testing methods are conventional in the art.

[0043] The cells were HCT116 (colon cancer cells), A549 (lung cancer cells), A375 (melanoma cells), Hela (cervical cancer cells), MCF-7 (breast cancer cells), Jurkat (blood cancer cells), HepG2 (liver cancer cells), U87 (human astrocytoblastoma cells), PANC-1 (pancreatic cancer cells), or Hek293 (human renal epithelial cells). A549, MCF-7, A375, and PANC-1 cells were obtained from the American Type Culture Collection, HCT116 cells were obtained from Wuhan Pronocell Life Science Co., Ltd., U87 cells were obtained from the Feng Min research group at the School of Pharmacy, Sun Yat-sen University, and HepG2, Jurkat, Hela, and Hek293 cells were obtained from the Wan Guohui research group at the School of Pharmacy, Sun Yat-sen University.

[0044] In the following examples or tests, the cancer cells were cultured as follows:

[0045] Take the culture flask containing the cancer cells required for the experiment, discard the original culture medium, wash three times with 2mL of PBS, add 1mL of trypsin to digest for 30s, and add 2mL of culture medium to terminate the digestion. Transfer the cells to a centrifuge tube, centrifuge at 1000 rpm for 3min, discard the supernatant, and add 2mL of culture medium to mix by pipetting 10 times. Take 10μL of the cell suspension and count it in a cell counter; plate a 96-well plate, seeding 5000 cells per well. For each compound, calculate the required cell volume and cell well volume by three replicate wells. Calculate the required cell suspension volume by cell counting, dilute the required number of cells with culture medium, plate 5000 cells per well, and plate 100μL of culture medium. Mix by pipetting, seed the cells using a dispenser, and culture in an incubator. Label the wells.

[0046] Example 1

[0047] This example studies the effect of BOLD-100 combined with metyrapone or acetazolamide on the inhibitory activity of cancer cells. The specific process is as follows:

[0048] MTT colorimetric assay for detecting cancer cell proliferation inhibitory effects

[0049] 24 hours after cell plating, a certain amount of metyrapone was added to each well of the 96-well plate to a concentration of 200 μM in each well. BOLD-100 was then added and diluted stepwise to act on the cells. After 48 hours, 20 μL of MTT was added to each well of the 96-well plate using a dispenser (MTT concentration was 5 mg / mL in PBS). The plates were incubated at 37°C for 4 hours. After the incubation period, the MTT mixture was aspirated and DMSO was added at 130 μL / well using a dispenser. The plates were shaken on a horizontal shaker for 10 minutes to fully dissolve the formazan. The absorbance was measured at a wavelength of 490 nm using a microplate reader. The cell survival rate at each drug concentration was calculated as required and a scatter plot was drawn.

[0050] The specific composition of the composition of metyrapone and ruthenate is shown in Table 1. The molar ratio in the table represents the molar ratio of the corresponding metyrapone compound to trans-[tetrachlorobis(1H-indazole)ruthenate(III) salt]:

[0051] Table 1

[0052]

[0053] The survival rates of HCT116 colon cancer cells treated with BOLD-100, the nitrogen-containing heterocyclic compounds described in Examples 1 to 8, and their combination are shown in Table 2.

[0054] Table 2

[0055]

[0056] Example 2

[0057] This example studies the effect of BOLD-100 combined with metyrapone on the inhibitory activity of various cancer cells. The specific process is as follows:

[0058] After treating the cells with metyrapone for 48 hours, 20 μL of MTT was added to each well of a 96-well plate using a dispenser (MTT concentration was 5 mg / mL in PBS); the plates were incubated at 37°C for 4 hours. After the incubation period, the MTT mixture was aspirated and DMSO was added at a rate of 130 μL / well using a dispenser. The plates were shaken on a horizontal shaker for 10 minutes to allow the formazan to fully dissolve. The absorbance was measured at a wavelength of 490 nm using a microplate reader. The cell survival rate at various drug concentrations was calculated as required, and a scatter plot was plotted. The results of the effects of BOLD-100, metyrapone, and the combination of the two compounds on cancer cell proliferation are shown in Table 3.

[0059] Table 3

[0060]

[0061] As shown in Table 3, under normal physiological conditions (adding FBS to the empty culture medium can simulate the protein environment under physiological conditions), BOLD-100 has no significant inhibitory effect on human colon cancer cells, human liver cancer cells, human lung cancer cells, human cervical cancer cells, human breast cancer cells, human astrocytoblastoma cells, human melanoma cells, blood cancer, and pancreatic cancer cells. However, when metyrapone was added, BOLD-100 exhibited excellent inhibitory effects on all five of the above cancer cells, and the other ligands had similar effects.

[0062] Example 3

[0063] This example studies the toxic and side effects of a ruthenate composition, and the specific process is as follows:

[0064] HEK293 cells were plated in a 96-well plate at a density of 5000 cells / well in DMEM complete medium. After 24 hours, metyrapone, 2-methylthiophenylboronic acid or 2-aminophenol were added (fixed concentration of 200 μM per well), and BOLD-100 was added in a stepwise dilution. After the plate was placed in an incubator and treated for 48 hours, 20 μL of BOLD-100 was added to each well using a spray gun. MTT was added to a 96-well plate (MTT concentration was 5 mg / mL in PBS); incubated at 37°C for 4 hours. After the incubation time, the MTT mixture was aspirated and DMSO was added at 130 μL / well using a dispenser; the plate was shaken on a horizontal shaker for 10 minutes until formazan was fully dissolved. The absorbance was measured at a wavelength of 490 nm using a microplate reader. The cell survival rate at each drug concentration was calculated as required, and a scatter plot was drawn. The effects of BOLD-100, metyrapone, 2-methylthiophenylboronic acid or 2-aminophenol, and their combination on cancer cell proliferation were shown in Table 4.

[0065] Table 4

[0066] Cell type Normal human renal epithelial cells cell lines Hek293 <![CDATA[BOLD-100(IC 50 )]]> 81±6μM <![CDATA[BOLD-100(IC 50 )+Metyrapone]]> 37±2μM BOLD-100 (IC 50 ) + 2-methylthiophenylboronic acid 3±1.5μM <![CDATA[BOLD-100(IC 50 )+2-aminophenol]]> 10±2μM <![CDATA[美替拉酮(IC 50 )]]> 1463±6μM <![CDATA[2-甲硫基苯硼酸(IC 50 )]]> 1000±5μM <![CDATA[2-氨基苯酚(IC 50 )]]> 820±10μM

[0067] As can be seen from Table 4, compared with 2-methylthiophenylboronic acid and 2-aminophenol, metyrapone has the lowest toxicity alone and the widest therapeutic window. Moreover, after combination with BOLD-100, the toxicity is only enhanced by 2.2 times compared with the toxicity of BOLD-100 itself, which is much lower than the 27-fold toxicity enhancement of BOLD-100 combined with 2-methylthiophenylboronic acid or the 8.1-fold toxicity enhancement of BOLD-100 combined with 2-aminophenol. 50 It is close to the therapeutic concentration range and the therapeutic window is extremely narrow.

[0068] Example 4

[0069] This example studies the actual inhibitory effect of BOLD-100 combined with metyrapone in a mouse tumor model. The detection steps are as follows:

[0070] Establishment of mouse tumor model: 2 million colon cancer cells HCT116 suspended in PBS were subcutaneously injected into the dorsal flank of 5-7 week old female BALB / c-nu / nu (nude mice) to establish xenograft model. When the tumor volume reached about 50mm 3 At 3-4 days after tumor inoculation, mice were randomly divided into a control group (Vehicle, using castor oil) and a treatment group (BOLD-100 group, metyrapone (2-MT) ​​group, and combination group). The combination group was treated with BOLD-100 (10 mg / kg mouse body weight / day) combined with metyrapone (100 mg / kg mouse body weight / day) by intraperitoneal injection for 5 consecutive days a week. When the tumor volume grew to 500 mm, the mice were randomly divided into two groups: a control group (Vehicle, using castor oil) and a treatment group (BOLD-100 group, metyrapone (2-MT) ​​group, and combination group). 3 Afterwards, the mice were anesthetized and then sacrificed by cervical dislocation.

[0071] The results are as follows Figures 4 to 7 shown.

[0072] It can be seen that the tumor growth was not inhibited in the control group, the group receiving BOLD-100 alone, or the group receiving metyrapone alone; when BOLD-100 and metyrapone were used in combination, tumor growth was significantly inhibited.

[0073] Example 5

[0074] This example studies the UV-VIS spectra and absorption kinetic curves of BOLD-100 itself and after mixing with metyrapone or acetazolamide in PBS. The detection steps are as follows:

[0075] BOLD-100 was dissolved in PBS to a 200 μM stock solution, which was then added to a 96-well plate. Subsequently, 400 μM metyrapone or acetazolamide was added to each sample group. After a period of time, changes in the UV-VIS spectra relative to the BOLD-100 at the zero point were measured.

[0076] The results are as follows Figure 8 、 9 As shown, UV-VIS showed that metyrapone or acetazolamide could effectively inhibit the formation of multinuclear ruthenium aggregates (absorption peak near 600 nm) and enhance the anticancer activity.

[0077] The above embodiments are preferred implementation modes of the present invention, but the implementation modes of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications that do not deviate from the spirit and principles of the present invention should be considered as equivalent replacement methods and are included in the scope of protection of the present invention.

Claims

1. A ruthenate composition, characterized in that: Including trans-[tetrachlorobis( 1 H-indazole) ruthenate (III) salt] and a medicinal nitrogen-containing heterocyclic compound, wherein the medicinal nitrogen-containing heterocyclic compound comprises metyrapone and / or acetazolamide.

2. The ruthenate composition according to claim 1, wherein: The medicinal nitrogen-containing heterocyclic compound and trans-[tetrachlorobis( 1 The molar ratio of H-indazole)ruthenate (III) is (0.1-50):

1.

3. The ruthenate composition according to claim 1, wherein: The metyrapone and trans-[tetrachlorobis( 1 The molar ratio of H-indazole)ruthenate (III) is (0.1-50):

1.

4. The ruthenate composition according to claim 1, wherein: The acetazolamide and trans-[tetrachlorobis( 1 The molar ratio of H-indazole)ruthenate (III) is (0.1-50):

1.

5. The ruthenate composition according to claim 1, wherein: The trans-[tetrachlorobis( 1 The structural formula of [H-indazole) ruthenate (III) salt] is shown in Formula I: Wherein, R is selected from Na + 、 6. The ruthenate composition according to claim 5, characterized in that: The trans-[tetrachlorobis( 1 H-indazole)ruthenate(III)] includes and / or 7. The ruthenate composition according to claim 1, wherein: The ruthenate composition further comprises a pharmaceutically acceptable excipient.

8. The ruthenate composition according to claim 1, wherein: The dosage form of the ruthenate composition includes any one of tablets, capsules, pills, granules, oral solutions, oral suspensions, syrups, injections, sterile powders for injection, concentrated solutions for injection, inhalation preparations, aerosols, powder sprays or sprays.

9. Use of the ruthenate composition according to any one of claims 1 to 8 in the preparation of anti-tumor drugs.

10. The use according to claim 9, characterized in that: The tumor includes at least one of colorectal cancer, liver cancer, lung cancer, brain cancer, cervical cancer, breast cancer, melanoma, pancreatic cancer, multiple myeloma or blood cancer.

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