CN108299473B - Complex of copper and benzimidazole compound containing pyridine and application thereof - Google Patents

Abstract

The invention belongs to the technical field of medicine, and relates to novel copper complexes, complexes of copper and pyridine-containing benzimidazole compounds, compositions containing these complexes, and applications in preparing antitumor drugs. The compound structure of the present invention is shown in the following formula (I): wherein, R is selected from H, C1-C6 alkoxy, halogenated C1-C6 alkoxy, hydroxyl, amino, nitro, cyano; R ₁ is selected from H, C1-C6 alkyl, C1-C6 alkoxy, halogenated C1-C6 alkoxy, hydroxyl, amino, nitro, cyano; R ₂ is selected from H, substituted or unsubstituted C1-C6 alkyl , C1-C6 alkoxy; Described substituent is C1-C6 alkoxy, halogenated C1-C6 alkoxy; R ₃ is selected from H, C1-C6 alkyl, C1-C6 alkoxy. The complex of the present invention and the composition containing the complex can be used to prepare antitumor drugs.

Description

Complexes of copper and benzimidazoles containing pyridine and their applications

technical field

The present invention relates to novel copper complexes, complexes of copper and pyridine-containing benzimidazole compounds, compositions containing these complexes, and their application in the preparation of antitumor drugs.

Background technique

Malignant tumor is recognized as one of the most serious diseases to human health in the world, and its harm is second only to cardiovascular disease. Therefore, strengthening the research on tumor treatment, improving the effect of various existing treatment methods, reducing cancer mortality, reducing recurrence rate, and improving the quality of life of cancer patients have important social benefits and huge economic benefits. In tumor treatment, chemotherapy is one of the important methods of current tumor treatment. Among many chemotherapeutic drugs, platinum group compounds are a class of drugs with strong anti-tumor effect and broad anti-tumor spectrum. Since Rosenberg et al. discovered that cisplatin has antitumor activity in 1967, cisplatin, as a highly effective antitumor drug, has been widely used in the treatment of various cancers, such as testicular cancer, ovarian cancer, cervical cancer, Bladder cancer, lung cancer and head and neck cancer, etc. (Wong E, Giandomenico CM. Current status of platinum-based antitumor drugs. Chem Rev, 1999, 99(9): 2451-2466.). Subsequently, a series of platinum group compounds such as carboplatin, nedaplatin, oxaliplatin, lobaplatin, and cycloplatin have been successfully used clinically as antitumor metal drugs. used (Abu-SurrahAS, Kettunen M. Platinum group antitumor chemistry: design and development of new anticancer drugs complementary to cisplatin. Curr Med Chem, 2006, 13(11): 1337-57.), but due to the toxicity and resistance of platinum group compounds The development of new antitumor drugs with transition metal complexes has attracted more and more attention.

At the same time, copper, as a biologically necessary trace element, exists in the active sites of metalloproteins and metalloenzymes in organisms, and participates in maintaining normal physiological functions in organisms. It plays an important role in the formation of skin pigment and the deposition of skin pigment, and has a unique catalytic role in the redox system of the organism, and plays a key role in aging and cancer and its associated DNA damage caused by endogenous oxidants (Wang Feili , Chang Yanling, An Lirong, etc. Research progress of non-platinum metal anticancer compounds. Chemical Research and Application, 2003, 15(5): 612-616.), and copper is more toxic than non-essential trace elements such as platinum Therefore, copper metal complexes are expected to maintain the antitumor activity of platinum group compounds while having low toxicity.

Based on the synthesis of some benzimidazole compounds containing pyridine groups, the present invention synthesizes a series of complexes of copper and benzimidazole compounds containing pyridine, and these complexes have significant anti-tumor properties. It is expected to become a candidate drug for novel anti-tumor therapy.

SUMMARY OF THE INVENTION

The technical problem solved by the present invention is to provide a compound shown in formula I, its prodrug and drug active metabolite and its pharmaceutically acceptable salt, and its application in the preparation of antitumor drugs.

The compound structure of the present invention is shown in the following formula (I):

in,

R is selected from H, C1-C6 alkoxy, halogenated C1-C6 alkoxy, hydroxy, amino, nitro, cyano;

R ₁ is selected from H, C1-C6 alkyl, C1-C6 alkoxy, halogenated C1-C6 alkoxy, hydroxyl, amino, nitro, cyano;

R ₂ is selected from H, substituted or unsubstituted C1-C6 alkyl, C1-C6 alkoxy; the substituents are C1-C6 alkoxy, halogenated C1-C6 alkoxy;

R ₃ is selected from H, C1-C6 alkyl, C1-C6 alkoxy.

Preferably,

R is selected from H, C1-C4 alkoxy, halogenated C1-C4 alkoxy;

R ₁ is selected from H, C1-C4 alkyl, C1-C4 alkoxy;

R ₂ is selected from H, C1-C4 alkyl, substituted or unsubstituted C1-C4 alkoxy, and the substituent is C1-C4 alkoxy, halogenated C1-C4 alkoxy;

R ₃ is selected from H, C1-C4 alkyl, C1-C4 alkoxy.

further,

R is selected from H, methoxy, difluoromethoxy;

R ₁ is selected from H, methyl, methoxy;

R ₂ is selected from H, methyl, methoxy, 2-methoxyethoxy, 3-methoxypropoxy, 2,2,2-trifluoroethoxy;

_R3 is selected from H, methyl, methoxy.

"Pharmaceutically acceptable salt" refers to conventional acid or base addition salts formed with suitable non-toxic organic or inorganic acids or bases that retain the biological potency and properties of the compounds of formula I. Acid addition salts include hydrochloride, hydrobromide, hydroiodide, nitrate, phosphate, sulfate, perchlorate, thiocyanate, bisulfate, persulfate, borate, Formate, Acetate, Propionate, Valerate, Pivalate, Caproate, Heptanoate, Caprylate, Isooctanoate, Undecanoate, Laurate, Palmitic Acid Salt, Stearate, Oleate, Cyclopropionate, Oxalate, Malonate, Succinate, Maleate, Fumarate, Adipate, Azelaate, Acrylic Acid Salt, strawberry salt, crotonate, tiglic acid, itaconate, sorbate, cinnamate, glycolate, lactate, malate, tartrate, citrate, tartrate, Mandelate, Diphenylglycolate, Tropineate, Ascorbate, Gluconate, Glucoheptonate, Gluconate, Mannoate, Lactobate, Benzoate, Phthalate , terephthalate, furoate, nicotinate, isonicotinate, salicylate, acetylsalicylate, butyrate, gallate, caffeate, ferulate, picrate , camphorate, camphorsulfonate, mesylate, ethanesulfonate, propanesulfonate, benzenesulfonate, p-toluenesulfonate, p-aminobenzenesulfonate, sulfamate, taurine acid salt, 2-hydroxyethanesulfonate, glycinate, alanine, valine, leucine, isoleucine, phenylalanine, tryptophan, tyrosinate , aspartate, asparagine, glutamate, lysine, glutamine, methionine, serine, threonine, cysteine, proline salt, histidine, arginate, edetate, pyruvate, alpha-ketoglutarate, alginate, cyclopentane propionate, 3-phenylpropionate, 3- Cyclohexylpropionic acid, 2-naphthoate, 2-naphthalenesulfonate, pamoate, lauryl sulfate, glycerophosphate, lauryl sulfate, pectinate, etc. Base addition salts include ammonium salts, alkali metal salts such as sodium and potassium salts, alkaline earth metal salts such as calcium and magnesium salts, salts of organic bases such as dicyclohexylamine salts, N-methyl-D-glucosamine Salts, etc., and basic nitrogen-containing groups can be quaternized with reagents such as lower alkyl halides such as methyl, ethyl, propyl and butyl chloride, bromine and iodide; dioxane sulfate esters such as dimethyl sulfate, diethyl, dibutyl and dipentyl; long-chain halides such as decyl, lauryl, myristyl and stearoyl chloride, bromine and iodide; aralkanes base halides, such as benzyl and phenethyl bromides, etc. Preferred acids for forming acid addition salts include hydrochloric acid, p-toluenesulfonic acid, methanesulfonic acid, maleic acid, malic acid, picric acid, citric acid, p-aminobenzenesulfonic acid.

"Pharmaceutically acceptable" such as a pharmaceutically acceptable carrier, excipient, prodrug, etc., means one that is pharmacologically acceptable and that is not substantially toxic to the patient to which the particular compound is administered.

"Pharmaceutically active metabolite" refers to a metabolite of a compound of Formula I that is pharmaceutically acceptable and effective.

The present invention also relates to pharmaceutical compositions thereof comprising formula I or a stereoisomer or a pharmaceutically acceptable acid addition salt thereof and a pharmaceutically acceptable carrier.

Certain crystalline forms of the compounds may exist in polycrystalline forms, which are also included in the present invention. In addition, some of the compounds may form solvates with water (ie, hydrates) or common organic solvents, and such solvates are also intended to be included within the scope of the present invention.

The present invention includes within its scope prodrugs of the compounds of the present invention. Typically, such prodrugs will be functional derivatives of the present compounds that are readily converted to the desired compound in vivo. Thus, in the methods of treatment of the present invention, the term "administering" will encompass the treatment of a variety of described conditions with compounds specifically disclosed, or compounds that are not specifically disclosed but are converted to the specified compounds in a patient after administration.

The following reaction schemes outline the preparative procedures for preparing the compounds of the present invention.

reaction flow

The present invention also relates to the complex of copper and pyridine-containing benzimidazole compounds used as an antitumor drug for tumor treatment.

Treatment and dosage used

The various diseases and conditions to be treated described herein are well known and apparent to those skilled in the art. It will also be appreciated that those of skill in the art can treat patients currently suffering from a disease or disorder with therapeutically effective amounts of the compound, or to affect related diseases and disorders by prophylactic treatment of patients suffering from a disease or disorder.

The term "patient" as used herein refers to a warm-blooded animal, such as a mammal, that has a tumor. It should be understood that guinea pigs, dogs, cats, rats, mice, horses, cows, sheep and humans are examples of animals within the meaning of this term.

The term "therapeutically effective amount" as used herein refers to an amount effective to control tumor-related diseases and disorders. The term "control" is used to refer to all processes that can slow, interrupt, arrest or stop the development of the diseases and disorders described herein, not necessarily requiring complete elimination of all symptoms of the diseases and disorders.

A therapeutically effective amount can be readily determined by the attending diagnosing physician as one skilled in the art using routine techniques and observing the results obtained under similar circumstances. In determining the dosage of a therapeutically effective amount, the attending physician takes into account a number of factors, including but not limited to: the species of mammal; its size, age, and general health; the specific disease involved; the extent or complexity or severity of the disease individual patient response; particular compound administered; mode of administration; bioavailability properties of the administered formulation;

A therapeutically effective amount of the compound is expected to vary from about 0.001 milligrams per kilogram of body weight per day (mg/kg/day) to about 100 mg/kg/day. The preferred amount can be determined by one skilled in the art.

Such compounds may be administered in therapeutically effective amounts in any form or manner that renders the compounds bioavailable, including oral, inhalation, and parenteral routes, in effective treatment of patients suffering from the aforementioned diseases and disorders. For example, the compounds can be administered orally, by inhalation aerosol or dry powder, subcutaneously, intramuscularly, intravenously, transdermally, intranasally, rectally, topically, and the like. Oral or inhalation administration is generally preferred for the treatment of respiratory diseases such as asthma. Those skilled in the art of formulating can readily select the appropriate form and mode of administration depending on the particular properties of the compound selected, the condition of the disease or disorder to be treated, the stage of the disease or disorder, and other relevant circumstances.

The compounds of the present invention may be administered alone or in combination with pharmaceutically acceptable carriers or excipients in the form of pharmaceutical compositions, the proportions and properties of which are selected by the solubility and chemical properties of the selected compound, route of administration, as determined by standard pharmaceutical guidelines. The compounds of the present invention, while effective in themselves, may be formulated and administered in the form of their pharmaceutically acceptable salts, such as acid addition salts or base addition salts, for stability, ease of crystallization, improved solubility, etc. Purpose.

The present invention provides pharmaceutical compositions containing a therapeutically effective amount of a compound in admixture or otherwise in association with one or more pharmaceutically acceptable carriers or excipients.

Pharmaceutical compositions are prepared by methods well known in the pharmaceutical art. Carriers or excipients that can serve as a carrier or medium for the active ingredient can be solid, semi-solid or liquid materials. Suitable carriers or excipients are well known in the art. The pharmaceutical compositions may be adapted for oral, inhalation, parenteral or topical use, and may be administered to a patient in the form of tablets, capsules, aerosols, inhalants, suppositories, solutions, suspensions, and the like.

The compounds of the present invention may be administered orally, eg, with an inert diluent or with an edible carrier. They may be enclosed in capsules or compressed into tablets. For oral therapeutic administration, the compounds can be mixed with excipients and used in the form of tablets, troches, capsules, elixirs, suspensions, syrups, wafers, chewing gums and the like. These formulations should contain at least 4% of the compound of the present invention, the active ingredient, but may vary depending on the particular form, conveniently varying from 4% to about 70% by weight of the unit. The amount of compound present in the composition should be such that a suitable dosage can be obtained. Preferred compositions and formulations of the present invention can be determined by those skilled in the art.

Tablets, pills, capsules, lozenges and the like may also contain one or more of the following adjuvants: binders such as microcrystalline cellulose, tragacanth or gelatin; excipients such as starch or lactose, disintegrants For example, alginic acid, Primogel, cornstarch, etc.; lubricants such as magnesium stearate or Sterotex; glidants such as colloidal silicon dioxide; and sweeteners such as sucrose or saccharin or flavoring agents such as peppermint, methyl salicylate may be added Ester or orange spice. When the dosage unit form is a capsule, it may contain, in addition to materials of the above type, a liquid carrier such as polyethylene glycol or a fatty oil. Other dosage unit forms may contain other various materials that modify the physical form of the dosage unit, for example, as coatings. Thus, tablets or pills may be coated with sugar, gum or other enteric coating agents. A syrup may contain, in addition to the compound, sucrose as a sweetening agent and certain preservatives, dyes and colorants and flavorings. The materials used in the preparation of these various compositions should be pharmaceutically pure and non-toxic in the amounts used.

For parenteral therapeutic administration, the compounds of the present invention may be added to a solution or suspension. These formulations should contain at least 0.1% of the compound of the present invention, but may vary between 0.1 and about 50% by weight of the formulation. The amount of compound present in such compositions should be such that a suitable dosage can be obtained. Preferred compositions and formulations can be determined by those skilled in the art.

The compounds of the present invention may also be administered by inhalation, eg, by aerosol or dry powder. Release may be with a liquefied or compressed gas or with a suitable pumping system for dispensing the compounds of the invention or formulations thereof. Formulations for administration of the compounds by inhalation can be delivered in monophasic, biphasic or triphasic systems. For aerosol administration of compounds, a number of systems are available. Dry powder formulations are prepared by granulating or milling the compound to a suitable particle size, or by mixing the granulated or milled compound with a suitable carrier such as lactose and the like. The delivery system by inhalation includes the necessary containers, activators, valves, sub-containers, and the like. Preferred aerosol and dry powder formulations for administration by inhalation can be determined by those skilled in the art.

The compounds of the present invention may also be administered topically, in which case the formulations suitably contain a solution, ointment or gel base. The base may contain, for example, one or more of the following: petrolatum, lanolin, polyethylene glycols, beeswax, mineral oil, diluents such as water and alcohol, and emulsifiers and stabilizers. Topical formulations contain calcium compounds or The concentration of a pharmaceutically acceptable salt thereof may be from about 0.1 to about 10% w/v (weight per unit volume).

The solution or suspension may also contain one or more of the following adjuvants: sterile diluents such as water for injection, saline solution, fixed oils, polyethylene glycols, glycerol, propylene glycol or other synthetic solvents; antibacterial agents such as benzyl Alcohols or methylparaben; antioxidants such as vitamin C or sodium bisulfite; chelating agents such as EDTA; buffers such as acetate, citrate or phosphate and osmotic-adjusting agents such as chloride sodium or glucose. The parenteral preparation can be presented in ampoules, disposable syringes or multiple-dose vials made of glass or plastic.

Description of drawings

Figure 1 is the molecular structure diagram of the complex G-12.

Detailed ways

Example 1: Bis(μ-sulfate-O,O')bis{2-[(3,4-dimethoxy-2-pyridyl)methylthio-κN]-1H-benzimidazole-κN ³ } Preparation of copper(II)(G-11)

After completely dissolving 1.27 g (7.97 mmol) of copper sulfate pentahydrate in 30 mL of methanol, 2-[(3,4-dimethoxy-2-pyridyl)methylthio]-1H-benzoyl was added dropwise thereto. The 10 mL methanol solution of imidazole 2.40g (7.97mmol) was reacted at room temperature overnight, and TLC detected, after the reaction was complete, the methanol solution was distilled off under reduced pressure to obtain a dark green solid, which was bis(μ-sulfate)bis{2-[( 3,4-Dimethoxy-2-pyridyl)methylthio]-1H-benzimidazole-N,N"-ylidene}dicopper(II) 3.67 g (3.99 mmol). Yield 100.0% .mp: >250°C; HRMS: 916.965191 ([MH] ^- );

Example 2: Bis(μ-sulfate-O,O')bis{2-[(3,4-dimethoxy-2-pyridyl)methylthio-κN]-1H-benzimidazole-κN ³ } Preparation of copper(II) methanol solvate (G-12)

Take about 5mg of bis(μ-sulfate-O,O')bis{2-[(3,4-dimethoxy-2-pyridyl)methylthio-κN]-1H-benzimidazole-κN ³ } Combine two copper (II), completely dissolve in 5mL methanol, then add different amounts of acetone, and simultaneously prepare mixed solutions of methanol and acetone in different proportions. Among them, methanol is used as a good solvent, and acetone is used as a poor solvent. Put the prepared mixed solutions in different proportions in different small conical flasks that were previously rinsed with chromic acid solution and dried, and the inner wall is smooth. The internal solvent evaporates slowly. Place the small Erlenmeyer flask in a quiet, dark, stable place for culture to get dark green crystals. Under a high magnification microscope, select regular and transparent single crystal particles, collect their crystal diffraction data on a Rigaku MSC X-ray crystal diffractometer, and use the SHELXL and SHELXS programs to analyze their crystal structures. The unit cell parameters are shown in Table 1. Its crystal structure see picture 1.

Table 1. Crystal structure parameters of complex G-12

Example 3: Bis(μ-sulfate-O,O')bis{5-difluoromethoxy-2-[(3,4-dimethoxy-2-pyridyl)methylthio-κN] Preparation of -1H-benzimidazole-κN ³ } copper(II)(G-13)

Referring to the preparation method of Example 1, dark green powder with metallic luster was obtained, and the yield was 100.0%. mp:>250℃; HRMS: 1050.919246([MH] ^- ); IR: 3422.7,3105.5,2949.5,2725.3,2169.5,1631.0,1501.4,1460.7,1432.1,1356.3,1309.5,1283.1,5.234.9.13 820.2.

Example 4: Bis(μ-sulfate-O,O')bis{2-{[3-methyl-4-(2,2,2-trifluoromethoxy)-2-pyridyl]methylsulfide Preparation of base-κN}-1H-benzimidazole-κN ³ } copper(II)(G-14)

Referring to the preparation method of Example 1, a lavender flake solid was obtained with a yield of 100.0%. mp:>250℃; HRMS: 1024.913695([MH] ^- ); IR: 3423.0, 2923.6, 2169.5, 1696.7, 1455.9, 1401.7, 1385.5, 1338.1, 1269.6, 1166.1, 1113.8, 1038.3, 960.8, 865.

Example 5: Bis(μ-sulfate-O,O')bis{5-methoxy-2-[(4-methoxy-3,5-dimethyl-2-pyridyl)methylthio Preparation of -κN]-1H-benzimidazole-κN ³ } copper(II)(G-15)

Referring to the preparation method of Example 1, dark green powder with metallic luster was obtained, and the yield was 100.0%. mp: >250℃; HRMS: 976.994612([MH] ^- ); IR: 3424.8, 2923.2, 2169.5, 1696.8, 1456.7, 1400.8, 1385.1, 1338.6, 1272.7, 1110.0, 1053.5, 1009.1, 865.2, 834.7.

Example 6: Bis(μ-sulfate-O,O')bis{2-[(4-methoxy-3,5-dimethyl-2-pyridyl)methylthio-κN]-1H- Preparation of benzimidazole-κN ³ } copper(II)(G-16)

Referring to the preparation method of Example 1, dark green powder with metallic luster was obtained, and the yield was 100.0%. mp:>250℃; HRMS: 914.973830([MH] ^- ); IR: 3422.8, 2923.6, 2169.5, 1696.6, 1451.4, 1385.3, 1354.6, 1274.1, 1087.1, 1039.1, 995.4, 865.4, 835.0.

Example 7: Bis(μ-sulfate-O,O')bis{5-difluoromethoxy-2-[(4-methoxy-3,5-dimethyl-2-pyridyl)methyl Preparation of thio-κN]-1H-benzimidazole-κN ³ } copper(II)(G-17)

Referring to the preparation method of Example 1, dark green powder with metallic luster was obtained, and the yield was 100.0%. mp: >250°C; HRMS: 1046.958230 ([MH] ^- ); IR: 3422.4, 2923.9, 2169.5, 1696.8, 1457.0, 1385.5, 1338.9, 1272.7, 1124.6, 1039.0, 1007.6, 865.3, 834.7.

Example 8: Bis(μ-sulfate-O,O')bis{5-methoxy-2-{[4-(2-methoxypropoxy)-3,5-dimethyl-2- Preparation of pyridyl]methylthio-κN}-1H-benzimidazole-κN ³ } copper(II)(G-18)

Referring to the preparation method of Example 1, dark green powder with metallic luster was obtained, and the yield was 100.0%. mp: >250°C; HRMS: 1063.050545 ([MH] ^- ); IR: 3422.1, 2924.0, 2169.5, 1696.6, 1458.3, 1338.2, 1273.7, 1159.2, 1096.0, 1037.5, 865.0, 834.1.

Example 9: Bis(μ-sulfate-O,O')bis{5-methoxy-2-{[2-methyl-4-(2,2,2-trifluoromethyl)-2- Preparation of pyridyl]methylthio-κN}-1H-benzimidazole-κN ³ } copper(II)(G-19)

Referring to the preparation method of Example 1, dark green powder with metallic luster was obtained, and the yield was 100.0%. mp: >250°C; HRMS: 1082.939430 ([MH] ^- ); IR: 3424.2, 2924.1, 2169.5, 1697.0, 1457.2, 1418.1, 1262.5, 1162.7, 1036.9, 963.8, 863.2, 834.3.

Example 10: Bis(μ-sulfate-O,O')bis{5-methoxy-2-[(3,4-dimethoxy-2-pyridyl)methylthio-κN]-1H - Preparation of benzimidazole-κN ³ } copper(II)(G-20)

Referring to the preparation method of Example 1, dark green powder with metallic luster was obtained, and the yield was 100.0%. mp: >250℃; HRMS: 978.956264 ([MH] ^- );

Example 11: Inhibitory activity of test compounds on the proliferation of human hepatoma cells HepG2, human colon cancer cells SW480 and human normal hepatocytes HL7702

(1) Experimental materials

Cell lines: human hepatoma cell HepG2, human colon cancer cell SW480, human normal cancer cell HL7702; 10% newborn bovine serum (FBS); 96-well plate; Dulbecco's modified Eagle's medium (DMEM).

Test compounds: L-01~L-16, C-01~C-16, a total of 32 compounds, dissolved in DMSO to 50mM and stored at -20℃ for later use, the final concentration of DMSO in the culture medium is less than 0.1% .

MTT experiment: dissolve in PBS to 5 mg/mL and store at -20°C.

(2) Experimental method

The antitumor activity of the test compounds was determined by MTT method (Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods. 1983, 65(1-2):55-63.).

HepG2 (liver cancer), SW480 (human colon cancer) and HL7702 (human normal hepatocytes) were cultured in Eagle's medium (DMEM). The medium is Dulbecco's modified and contains 10% fetal bovine serum (FBS), SW480 cell line and HepG2 cell line. Each compound was administered at final concentrations of 50 uM and 25 uM, with 3 parallel wells for each concentration. When the cells proliferated to 80-90%, they were pooled and then subcultured for no more than 20 passages, and then they were acclimated to the environment for 24 h before the next treatment. The cells were plated in 96-well plates ( ⁸ x 104/mL) and then cultured overnight in a humidified environment containing 5% _CO2 and temperature-controlled at 37°C. After culturing at 37°C for 24 hours, add human MTT (5mg/ml) solution to each well, shake the culture plate gently, put it back into the incubator for another 4 hours, then suck up the supernatant, add 100 μL of DMSO to each well, Shake on a shaker for 5-10 min to completely dissolve MTT, and measure the absorbance value (OD value) at a wavelength of 490 nm in each well with an enzyme label luminometer (TECAN SPECTRA, Wetzlar, Germany), and calculate the inhibition rate with the following formula (see Table 2).

Inhibition rate=(1-OD value of drug group/OD value of control group)×100%

Cisplatin was used as the positive control drug in the experiment. The IC ₅₀ of cisplatin on the proliferation of human hepatoma cells (HepG2) and human colon cancer cells (SW480) were 6.6±1.0μmol/L and 15±2.7μmol/L, respectively.

The experimental results show that the tested copper complexes have different inhibitory effects on the proliferation of human hepatoma cell HepG2, and show a dose-effect relationship. Only a few tested copper complexes have inhibitory effects on the proliferation of human colon cancer cell SW480. The inhibitory effect of some copper complexes on the proliferation of human hepatoma cell HepG2 is stronger than that on human colon cancer cell SW480, while the tested copper complexes hardly inhibit the proliferation of human normal hepatocyte HL7702 effect.

Table 2. Inhibitory effects of test compounds on the proliferation of various cells

pharmaceutical composition

Example 13: Tablet Formulation

Active compound 25-1000 mg, starch 45 mg, microcrystalline cellulose 35 mg, polyvinylpyrrolidone (as a 10% aqueous solution) 4 mL, sodium carboxymethyl cellulose 4.5 mg, magnesium stearate 0.5 mg, talc 1 mg.

Example 14: Suspension Agent Formulation

Active compound 0.1-1000 mg, sodium carboxymethyl cellulose 50 mg, syrup 1.25 mg, sodium benzoate 0.1 mg, appropriate amount of flavoring agent, appropriate amount of coloring agent, add purified water to 5 mL.

Example 15: Aerosol Formulation

Active compound 0.25 mg, ethanol 25-75 mL, propellant 22 (chlorodifluoromethane) 70 mg.

Example 16: Suppository Formulation

Active compound 250mg, saturated fatty acid glycerides 2000mL.

Example 17: Injectable Formulation Formulation

Active compound 50 mg, isotonic saline solution 1000 mL.

Example 18: Ointment formulation

Micronized active compound 0.025g, liquid paraffin 10g, add soft white wax to 100g.

Example 19: Ointment formulation

Active compound 0.025g, propylene glycol 5g, sorbitan sesquioleate 5g, liquid paraffin 10g, add soft white wax to 100g.

Example 20: Oil-in-water cream formulation

Active compound 0.025g, cetyl alcohol 5g, glycerol monostearate 5g, liquid paraffin 10g, Cetomacrogol1 000 2g, citric acid 0.1g, sodium citrate 0.2g, propylene glycol 35g, add water to 100g.

Example 21: Oil-in-water cream formulation

Micronized Active Compound 0.025g, Soft White Wax 15g, Liquid Paraffin 5g, Cetyl Alcohol 5g, Sorbimacrogolstearate 2g, Sorbitan Monostearate 0.5g, Sorbic Acid 0.2g, Citric Acid 0.1g, Sodium Citrate 0.2g, Add water to 100g.

Example 22: Water-in-Oil Cream Formulation

Active compound 0.025g, soft white wax 35g, liquid paraffin 5g, sorbitan sesquioleate 5g, sorbic acid 0.2g, citric acid 0.1g, sodium citrate 0.2g, add water to 100g.

Example 23: Lotion formulation

Active compound 0.25g, isopropanol 0.5mL, carboxyvinyl polymer 3mg, appropriate amount of NaOH, add water to 1g.

Example 24: Suspension formulation for injection

Active compound 0.05-10 mg, sodium carboxymethyl cellulose 7 mg, NaCl 7 mg, polyoxyethylene (20) sorbitan monooleate 0.5 mg, benzyl alcohol 8 mg, add sterile water to 1 mL.

Example 25: Aerosol formulation for oral and nasal inhalation

Active Compounds 0.1%w/w, Sorbitan Trioleate 0.7%w/w, Trichlorofluoromethane 24.8%w/w, Dichlorotetrafluoroethane 24.8%w/w, Dichlorodifluoromethane 49.6% w/w.

Example 26: Atomized Solution Formulation

Active compound 7 mg, propylene glycol 5 mg, and water to 10 g.

Example 27: Powder formulation for inhalation

A gelatin capsule is filled with a mixture of the following ingredients, micronized active compound 0.1 mg, lactose 20 mg, and the powder is inhaled by means of an inhalation device.

Example 28: Powder formulation for inhalation

The spheronized powder is filled into a multi-dose powder inhaler, each dose containing 0.1 mg of micronized active compound.

Example 29: Powder formulation for inhalation

The spheronized powder is filled into a multi-dose powder inhaler, each dose containing 0.1 mg of micronized active compound and 1 mg of micronized lactose.

Example 30: Capsule Formulation

Active Compound 1.0 Sugar Balls 321mg, Aquacoat ECD 30 6.6mg, Acetyl Tributyl Citrate 0.5mg, Tween-80 0.1mg, Eudragit L 100-55 17.5mg, Triethyl Citrate 1.8mg, Talc 8.8mg , Defoamer MMS 0.1mg.

Example 31: Capsule Seedling Formulation

Active Compounds 2.0mg, Sugar Balls 305mg, Aquocoat ECD 30 5.0mg, Acetyl Tributyl Citrate 0.4mg, Tween-80 0.14mg, Eudragit NE30D 12.6mg, Eudragit S100 12.6mg, Talc 0.16mg.

Example 32: Enema Formulation

Active Compounds 00.2mg, Sodium Carboxymethylcellulose 25mg, Disodium EDTA 0.5mg, Methylparaben 0.8mg, Propylparaben 0.2mg, Sodium Chloride 7mg, Citric Acid 1.8mg , Tween-80 0.01mg, add purified water to 1mL.

Example 33: Formulation containing liposomes

A. Preparation of instillation formulations

Combine synthetic dipalmitoyl lecithin (45 mg), dimyristoyl lecithin (7 mg), dipalmitoyl phosphatidyl glycerol (1 mg) and active compound (5 mg) in a glass tube and dissolve all components in chloroform _In , most of the solvent was evaporated with N, followed by reduced pressure, whereby a lipid film was formed on the surface of the glass tube. An aqueous solution (0.9% NaCl) was added to the lipid, at a temperature higher than the lipid inversion temperature Liposomes were formed and the resulting suspension contained liposomes ranging in size from very small vesicles to 2 μm.

B. Preparation of Formulations for Inhalation

Liposomes were prepared according to Example A, wherein the aqueous solution contained 10% lactose, and the ratio of lactose to lipid was 7:3. The liposome suspension was frozen on dry ice and freeze-dried, and the dried product was micronized to obtain particles with a mass mean aerodynamic diameter (MMAD) of about 2 μm.

The above are only preferred embodiments of the present invention, and are not intended to limit the present invention in other forms. Any person skilled in the art may use the technical content disclosed above to make changes or modifications to equivalent changes. Example. Any simple modifications, equivalent changes and modifications made to the above embodiments according to the technical essence of the present invention without departing from the content of the technical solutions of the present invention still belong to the protection scope of the technical solutions of the present invention.

Claims (27)

1. A compound or pharmaceutically acceptable salt of formula (I):

wherein,

r is selected from H, C1-C6 alkoxy, halogenated C1-C6 alkoxy;

R₁selected from H, C1-C6 alkyl, C1-C6 alkoxy, halogenated C1-C6 alkoxy;

R₂selected from H, substituted or unsubstituted C1-C6 alkyl, C1-C6 alkoxy; the substituent is C1-C6 alkoxy, halogenated C1-C6 alkylAn oxy group;

R₃selected from H, C1-C6 alkyl, C1-C6 alkoxy.

2. The compound or pharmaceutically acceptable salt of claim 1:

r is selected from H, C1-C4 alkoxy, halogenated C1-C4 alkoxy.

3. The compound or pharmaceutically acceptable salt of claim 1:

r is selected from H, methoxy and difluoromethoxy.

4. The compound or pharmaceutically acceptable salt of any one of claims 1-3:

R₁selected from H, C1-C4 alkyl, C1-C4 alkoxy.

5. The compound or pharmaceutically acceptable salt of any one of claims 1-3:

R₁selected from H, methyl and methoxy.

6. The compound or pharmaceutically acceptable salt of claim 4:

R₁selected from H, methyl and methoxy.

7. The compound or pharmaceutically acceptable salt of any one of claims 1-3:

R₂is selected from H, substituted or unsubstituted C1-C4 alkyl, C1-C4 alkoxy, and the substituent is C1-C4 alkoxy and halogenated C1-C4 alkoxy.

8. The compound or pharmaceutically acceptable salt of claim 4:

R₂is selected from H, substituted or unsubstituted C1-C4 alkyl, C1-C4 alkoxy, and the substituent is C1-C4 alkoxy and halogenated C1-C4 alkoxy.

9. The compound or pharmaceutically acceptable salt of claim 5:

R₂is selected from H, substituted or unsubstituted C1-C4 alkyl, C1-C4 alkoxy, and the substituent is C1-C4 alkoxy and halogenated C1-C4 alkoxy.

10. The compound or pharmaceutically acceptable salt of any one of claims 1, 2, 3, or 6:

R₂selected from H, methyl, methoxy, 2-methoxyethoxy, 3-methoxypropoxy, 2,2, 2-trifluoroethoxy.

11. The compound or pharmaceutically acceptable salt of claim 4:

R₂selected from H, methyl, methoxy, 2-methoxyethoxy, 3-methoxypropoxy, 2,2, 2-trifluoroethoxy.

12. The compound or pharmaceutically acceptable salt of claim 5:

R₂selected from H, methyl, methoxy, 2-methoxyethoxy, 3-methoxypropoxy, 2,2, 2-trifluoroethoxy.

13. The compound or pharmaceutically acceptable salt of any one of claims 1, 2, 3,6, 8, 9, 11, or 12:

R₃selected from H, C1-C4 alkyl, C1-C4 alkoxy.

14. The compound or pharmaceutically acceptable salt of claim 4:

R₃selected from H, C1-C4 alkyl, C1-C4 alkoxy.

15. The compound or pharmaceutically acceptable salt of claim 5:

R₃selected from H, C1-C4 alkyl, C1-C4 alkoxy.

16. The compound or pharmaceutically acceptable salt of claim 7:

R₃selected from H, C1-C4 alkyl, C1-C4 alkoxy.

17. The compound or pharmaceutically acceptable salt of claim 10:

R₃selected from H, C1-C4 alkyl, C1-C4 alkoxy.

18. The compound or pharmaceutically acceptable salt of any one of claims 1, 2, 3,6, 8, 9, 11, or 12:

R₃selected from H, methyl and methoxy.

19. The compound or pharmaceutically acceptable salt of claim 4:

R₃selected from H, methyl and methoxy.

20. The compound or pharmaceutically acceptable salt of claim 5:

R₃selected from H, methyl and methoxy.

21. The compound or pharmaceutically acceptable salt of claim 7:

R₃selected from H, methyl and methoxy.

22. The compound or pharmaceutically acceptable salt of claim 10:

R₃selected from H, methyl and methoxy.

23. A compound or pharmaceutically acceptable salt of:

bis (. mu. -sulfate-O, O ') bis {2- [ (3, 4-dimethoxy-2-pyridyl) methylthio ] -1H-benzimidazol-N, N ' ' -ylidene } cupro-nium;

bis (. mu. -sulfate-O, O ') bis {2- [ (3, 4-dimethoxy-2-pyridyl) methylthio ] -1H-benzimidazol-N, N ' ' -ylidene } cupro-methanol solvate;

bis (. mu. -sulfate-O, O ') bis { 5-difluoromethoxy-2- [ (3, 4-dimethoxy-2-pyridyl) methylthio ] -1H-benzimidazol-N, N' -ylidene } cupra;

bis (μ -sulfate-O, O') bis {2- { [ 3-methyl-4- (2,2, 2-trifluoromethoxy) -2-pyridyl ] methylthio } -1H-benzoimidazol-N, N "-ylidene } cupro-nium;

bis (. mu. -sulfate-O, O ') bis { 5-methoxy-2- [ (4-methoxy-3, 5-dimethyl-2-pyridyl) methylthio ] -1H-benzimidazol-N, N' -ylidene } cupro-bium;

bis (. mu. -sulfate-O, O ') bis {2- [ (4-methoxy-3, 5-dimethyl-2-pyridyl) methylthio ] -1H-benzimidazol-N, N' -ylidene } cupro-nium;

bis (μ -sulfate-O, O') bis { 5-difluoromethoxy-2- [ (4-methoxy-3, 5-dimethyl-2-pyridyl) methylthio ] -1H-benzimidazol-N, N "-ylidene } cupro-di;

bis (. mu. -sulfate-O, O ') bis { 5-methoxy-2- { [4- (2-methoxypropoxy) -3, 5-dimethyl-2-pyridinyl ] methylthio } -1H-benzimidazol-N, N' -ylidene } cupro-nium;

bis (μ -sulfate-O, O') bis { 5-methoxy-2- { [ 2-methyl-4- (2,2, 2-trifluoromethyl) -2-pyridinyl ] methylthio } -1H-benzimidazol-N, N "-ylidene } cupro-nium;

bis (. mu. -sulfate-O, O ') bis { 5-methoxy-2- [ (3, 4-dimethoxy-2-pyridyl) methylthio ] -1H-benzimidazol-N, N' -ylidene } cupro-biscopper.

24. A process for preparing a compound or pharmaceutically acceptable salt according to claim 1,

。

25. a pharmaceutical composition comprising as active ingredient a compound or pharmaceutically acceptable salt according to any one of claims 1-23 and a pharmaceutically acceptable carrier or excipient.

26. Use of a compound or pharmaceutically acceptable salt according to any one of claims 1 to 23 or a pharmaceutical composition according to claim 25 for the manufacture of a medicament for the treatment of a tumour.

27. Use according to claim 26, characterized in that: the tumor is liver cancer, colon cancer or rectal cancer.

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