KR20040008410A - Novel 5'-deoxy-n-alkyloxycarbonyl-5-fluorocytosine-5'-thioester derivatives, their preparation, and anticancer agent comprising the same - Google Patents

Abstract

PURPOSE: 5'-Deoxy-N-alkyloxycarbonyl-5-fluorocytosine-5'-thioester derivatives, a preparation method thereof, an anticancer agent containing the same as effective components are provided. The compounds have improved anticancer effect and low toxicity. CONSTITUTION: 5'-deoxy-N-alkyloxycarbonyl-5-fluorocytosine-5'-thioester derivatives are represented by the formula(1), wherein R1 is hydrogen, or C1-C10 saturated or unsaturated carbohydrate; R2 is hydrogen, or atom group easily hydrolyzed under physiological condition or easily removable protecting group; and R3 is C2-C7 saturated or unsaturated carbohydrate. A method for preparing the 5'-deoxy-N -alkyloxycarbonyl-5-fluorocytosine-5'-thioester derivatives of the formula(1) comprises the steps of: (a) reacting a compound of the formula(2) with thiol of the formula(3) of R1-SH in the presence of a solvent to prepare a compound of the formula(1b); and (b) deprotecting the compound of the formula 1b.

Description

Novel 5'-deoxy-N-alkyloxycarbonyl-5-fluorocytosine-5'-thioester derivative, preparation method thereof, and anticancer agent comprising the same as an active ingredient {NOVEL 5'-DEOXY-N-ALKYLOXYCARBONYL -5-FLUOROCYTOSINE-5'-THIOESTER DERIVATIVES, THEIR PREPARATION, AND ANTICANCER AGENT COMPRISING THE SAME}

The present invention relates to a novel 5'-deoxy-N-alkyloxycarbonyl-5-fluorocytosine-5'-thioester derivative, a preparation method thereof, and an anticancer agent comprising the same as an active ingredient, in more detail. Is a novel 5'-deoxy-N-alkyloxycarbonyl-5-fluorocytosine-5'-thioester derivative which can be used effectively as a new anticancer agent due to its excellent anticancer effect, a preparation method thereof, and an active ingredient thereof It relates to an anticancer agent comprising.

Cancer, along with AIDS, is one of the incurable diseases that modern medicine remains to address. In Korea, the incidence of cancer is increasing year by year, and cancer is a major cause of death due to its low healing rate, and the type and frequency of cancer are increasing day by day due to changes in modern society. Although the world is making huge investments to cure cancer, no satisfactory treatment has yet been developed. Therefore, it is essential to develop a cancer drug having excellent drug efficacy and fewer side effects for more efficient and effective healing.

Currently, as a method used to treat cancer, local therapies such as surgery, radiation therapy, and systemic therapy such as chemotherapy and immunotherapy are used. Among them, chemotherapy is widely used for the treatment of various cancers, such as solid tumors that occur primarily in various organs for gastrointestinal, liver, lung, etc. have.

In addition, research on cancer has been carried out by many scholars from several points of view, including 5-Fluorouracil (5-FU), Methotrexate, Frutraful, Cisplatin, etc. In addition to the development of anticancer drugs, research is being continued on new anticancer drugs. However, no formulations have yet been developed that can cure completely safely.

Therefore, there is a growing interest in developing a new anticancer agent having low toxicity and excellent effect, and recently using a derivative of 5-fluorouracil (5-FU), a representative anticancer agent of pyrimidine nucleoside family, Anticancer drug research is actively being conducted. However, 5-FU, which is widely known as an anticancer agent, is used for the treatment of tumors by metabolizing and activating in cells to inhibit the synthesis of pyrimidine nucleotides.

Therefore, many researches are being conducted to reduce this, but some of the developed 5-FU derivatives are activated in the intestinal wall after oral administration and cause side effects such as amputation, diarrhea and abdominal pain.

Recently, N-alkyloxycarbonyl 5'-deoxy-5-fluorocytidine represented by the following general formula (I) in European Patent No. 602454, Japanese Patent Laid-Open No. 6-211891 and US Patent No. 5,472,949 Processes for preparing derivatives are described.

(Wherein R ^a is a saturated or unsaturated hydrocarbon, R ^b represents hydrogen or an atomic group and easily removable protecting group that can be easily hydrolyzed under physiological conditions)

Said document is characterized by the fact that the 4'- position of cytidine system is a methyl group. However, the derivatives are known to be activated by enzymes in the liver, not in the intestine, and the side effects are greatly reduced. However, the derivatives are not yet completely able to safely cure cancer.

Therefore, in view of the above problems, the present invention is a novel 5'-deoxy-N-alkyloxycarbonyl-5-fluorocytosine-5'-thio compound which is a novel compound having superior anticancer effect compared to conventional anticancer agents. It is an object to provide an ester derivative.

Another object of the present invention is to provide a method for preparing the novel 5'-deoxy-N-alkyloxycarbonyl-5-fluorocytosine-5'-thioester derivative.

Another object of the present invention is to provide a novel 5'-deoxy-N-alkyloxycarbonyl-5-fluorocytosine-5'-thioester derivative, a pharmaceutically acceptable salt thereof, or a solvate thereof. It is to provide an anticancer agent comprising.

In order to achieve the above object, the present invention is a 5'-deoxy-N-alkyloxycarbonyl-5-fluorocytosine-5'- thioester derivative represented by the following formula (1), pharmaceutically acceptable salts thereof Or solvates thereof.

[Formula 1]

Where

R ₁ is hydrogen or C ₁ -C ₁₀ saturated or unsaturated hydrocarbon; R ₂ is hydrogen or an atomic group or easily removable protecting group that can be readily hydrolyzed under physiological conditions; R ₃ is a C ₂ -C ₇ saturated or unsaturated hydrocarbon.

In addition, the present invention

a) combining the compound of Formula 2 with a thiol of Formula 3 in the presence of a solvent to prepare a compound of Formula 1b; And

b) deprotecting the compound of Formula 1b to prepare a compound of Formula 1a

It provides a method for producing a compound of Formula 1 comprising a.

[Formula 1a]

[Formula 1b]

[Formula 2]

[Formula 3]

R ₁ -SH

Where

R ₁ is hydrogen or C ₁ -C ₁₀ saturated or unsaturated hydrocarbon; R ₂ is hydrogen or an atomic group or easily removable protecting group that can be readily hydrolyzed under physiological conditions; R ₃ is a C ₂ -C ₇ saturated or unsaturated hydrocarbon.

In addition, the present invention provides an anticancer composition comprising the compound of Formula 1, a pharmaceutically acceptable salt thereof, or a solvate thereof as an active ingredient.

Hereinafter, the present invention will be described in detail.

The present inventors studied a wide range of 5-FU series compounds and derivatives thereof to develop excellent anticancer agents, and converted to thioester derivatives instead of methyl groups at the 4'-position of the 5-FU series compounds and derivatives thereof. The 5'-deoxy-N-alkyloxycarbonyl-5-fluorocytosine-5'-thioester derivatives were confirmed to have excellent anticancer effects, and thus the present invention was completed.

Referring to the compound of Formula 1 of the present invention in more detail based on the preparation method as follows.

In the compound of Formula 1 of the present invention, R ₂ is hydrogen or an atomic group capable of easily hydrolyzing under physiological conditions by having a functional group such as acetyl, or an easily removable protecting group.

More preferably, the compound of Formula 1 is a compound of Formula 1a wherein R ₂ is hydrogen. The compound of Formula 1a may be prepared by reducing the compound of Formula 1b through a deprotection reaction in the presence of a solvent.

[Formula 1a]

[Formula 1b]

In which R ₁ is hydrogen or C ₁ -C ₁₀ saturated or unsaturated hydrocarbon; R ₃ is a C ₂ -C ₇ saturated or unsaturated hydrocarbon.

The compound of Formula 1b may be prepared by combining the compound of Formula 2 with the compound of Formula 3 in the presence of a solvent.

In the present invention, the compound of Formula 2 may be prepared through a protection reaction from a compound of Formula 4 (Korean Patent Application No. 2000-0046179).

[Formula 4]

Wherein R ₃ is a C ₂ -C ₇ saturated or unsaturated hydrocarbon.

For example of the production method of the compound of Formula 1 of the present invention, the following scheme 1.

Scheme 1

Wherein R ₁ and R ₃ are each as defined above.

Referring to Scheme 1, Compound 2 may be prepared by refluxing a p-toluenesulfonic acid catalyst and 2,2-dimethoxypropane to a compound of Formula 4 under acetone solvent conditions. to the next. Compound 1b may be prepared by combining Compound 2 with a thiol of Formula 3 (saturated or unsaturated hydrocarbon of R ₁ = C ₁ -C ₁₀ ) using a dimethylaminopyridine catalyst and a dicyclohexylcarbodiimide (DCC). have. Chloroform, methylene chloride, carbon tetrachloride and the like may be used as the solvent in this reaction. Next, Compound 1a can be prepared by deprotecting the Compound 1b under acidic conditions. In this reaction, trichloroacetic acid is mainly used.

In addition, the present invention may provide a pharmaceutical composition comprising the compound of Formula 1, a pharmaceutically acceptable salt thereof, or solvate thereof as an active ingredient. The pharmaceutical composition is preferably an anticancer agent.

The compounds of the present invention can be administered by various oral or parenteral methods. The pharmaceutical composition of the present invention may comprise a pharmacologically acceptable liquid or solid carrier.

The solid form preparations include powders, tablets, dispersible granules or capsules, among which solid dosage forms suitable for oral administration include tablets, powders or capsules. Suitable excipients may include diluents, flavors, solubilizers, lubricants, suspending agents, binders, and / or tableting agents. In the case of powders or capsules, the carrier may contain 5 to 70%, preferably 10 to 70%, of the finely divided active ingredient. Suitable solid carriers or excipients include corn starch, magnesium stearate, film, polyethylene glycol, talc, sugar, lactose, pectin, dextrin, starch, gelatin, hydroxypropylmethylcellulose, methylcellulose, sodium carboxymethylcellulose, titanium dioxide, low Melting point wax, cocoa butter and the like.

The preparations in liquid form may be solutions, suspensions or emulsions. For example, in the case of parenteral injection, water or a mixed solution of water-propylene glycol may be used, and such a solution is prepared such that isotonicity, pH, and the like are suitable for a biological system. Liquid formulations may also be formed from aqueous polyethyleneglycol solution. Aqueous solutions suitable for oral use may be prepared by dissolving the active ingredient in water and adding the appropriate flavoring, coloring, stabilizing and thickening agents. Aqueous suspensions suitable for oral use can be prepared by dispersing the finely divided active component in a viscous material such as natural or synthetic gums, resins, methylcellulose, sodium carboxymethylcellulose and known suspending agents.

Preferred pharmaceutical preparations are in unit dosage form. In such forms, the preparation is subdivided into unit dosage forms containing an appropriate amount of active ingredient. The unit dosage form can be a packaged preparation containing discrete amounts of the preparation, for example a packaged tablet, capsule or powder in a vial or ampoule.

Hereinafter, the present invention will be described in more detail with reference to Examples. However, the following examples are provided to aid the understanding of the present invention, and the scope of the present invention is not limited thereto.

EXAMPLE

Example 1

(5'-deoxy-2 ', 3'-(O-isopropylidene) -5-fluoro-N ^4- (pentyloxycarbonyl) -cytosine-5'-carboxylic acid (R ₃ = pentyl , Preparation of compound 2)

To 10 g (23.7 mmol) of 5'-deoxy-5-fluoro-N ^4- (pentyloxycarbonyl) -cytosine-5'-carboxylic acid (R ₃ = pentyl, compound 4) with 50 ml of acetone and Excess 2,2-dimethoxypropane and 0.05 equivalents of p-toluenesulfonic acid were added and refluxed for 3 hours. Then, the solvent was distilled under reduced pressure, washed with water, extracted with ethyl acetate, dried over anhydrous magnesium sulfate, filtered and concentrated under reduced pressure to give 10.7 g (yield 98%) of the title compound.

¹ H NMR (CDCl ₃ , ppm) δ 8.05 (br.s, 1H), 6.17 (d, 1H), 4.95-4.54 (m, 3H), 4.09 (t, 2H), 1.58 (m, 2H), 1.43 (s, 3H), 1.41 (s, 3H), 1.33 (m, 4H), 0.89 (t, 3H)

Example 2

(5'-deoxy-2 ', 3'-(O-isopropylidene) -5-fluoro-N ^4- (pentyloxycarbonyl) -cytosine-5'-thiolic acid phenyl (R ₁ = phenyl , R ₃ = pentyl, preparation of compound 1b)

5'-deoxy-2 ', 3'-(O-isopropylidene) -5-fluoro-N ^4- (pentyloxycarbonyl) -cytosine-5'-carboxylic acid (R ₃ at 0 ° C) = Pentyl, compound 2) 50 ml of methylene chloride, 1.5 equivalents of dicyclohexylcarbodiimide (DCC), 0.05 equivalents of dimethylaminopyridine, and 1.5 equivalents of thiophenol (R ₁ = phenyl, Compound 3) in 10.5 g (22.8 mmol) Were added sequentially and stirred at room temperature for about 10-20 hours, and then the solid was filtered off. Thereafter, the solution from which the solid was removed was concentrated by distillation under reduced pressure, and then purified by silica gel column chromatography to obtain 11.3 g (yield 95%) of the title compound.

¹ H NMR (CDCl ₃ , ppm) δ 8.06 (br.s, 1H), 7.10 (m, 5H), 6.16 (d, 1H), 4.95-4.44 (m, 3H), 4.09 (t, 2H), 1.58 (m, 2H), 1.43 (s, 3H), 1.41 (s, 3H), 1.35 (m, 4H), 0.90 (t, 3H)

Example 3

(Preparation of 5'-deoxy-5-fluoro-N ^4- (pentyloxycarbonyl) -cytosine-5'-thiolic acid phenyl (R ₁ = phenyl, R ₃ = pentyl, compound 1a))

5'-deoxy-2 ', 3'-(O-isopropylidene) -5-fluoro-N ^4- (pentyloxycarbonyl) -cytosine-5'-thiolic acid phenyl (R ₁ = phenyl, R ₃ = pentyl, compound 1b) 50 g of methylene chloride and an excess of trifluoroacetic acid were added to 11 g (21.1 mmol), and the mixture was stirred at room temperature for about 2 hours, followed by distillation under reduced pressure. Thereafter, ethanol was added and distillation under reduced pressure was performed several times to completely remove trifluoroacetic acid, and then 5.1 g (yield 50%) of the title compound was obtained by recrystallization.

¹ H NMR (CDCl ₃ , ppm) δ 8.05 (br.s, 1H), 7.10 (m, 5H), 6.16 (d, 1H), 4.70-4.28 (m, 3H), 4.09 (t, 2H), 1.58 (m, 2H), 1.35 (m, 4H), 0.89 (t, 3H)

Example 4

(Measurement of anticancer effect of derivatives)

Capecitabine = 5'-deoxy-5-fluor, showing the best results among the 5'-deoxy-N-alkyloxycarbonyl-5-fluorocytidine derivatives disclosed in European Patent No. 602454. Rho-N ^4- (pentyloxycarbonyl) cytidine (R ^a = pentyl, R ^b = hydrogen compound of general formula (I))] as a control agent, the compound according to the present invention 5'-deoxy- In order to measure the anticancer activity of 5-fluoro-N ^4- (alkyloxycarbonyl) -cytosine-5'-thioester, cytotoxicity against human cancer cells was measured. Human cancer cells were compared using A549 (lung cancer), HCT15 (colon cancer), SK-OV-3 (ovarian cancer), and SK-MEL-2 (skin cancer).

First, the cancer cells were incubated at 37 ° C. in a 5% CO ₂ thermohygrostat, and the medium used was 10% fetal bovine serum with RPMI as the basal medium. To measure the cytotoxic activity, the cancer cells in the logarithmic phase were divided into 2-5 × 10 ⁴ cells per well of a 96-well plate and incubated for 24 hours, followed by capecitabine and 5′-deoxy-5-fluoro. A sample solution of step-diluted rho-N ^4- (alkyloxycarbonyl) -cytosine-5'-thioester was added and incubated for 72 hours. 20 μl of MTT reaction solution dissolved in physiological saline at a concentration of 5 mg / ml in each well of the cultured plate was incubated for 4 hours, and the formed formazan crystals were dissolved in dimethyl sulfoxide (DMSO). The viable cell number was measured by measuring the absorbance of each well at a wavelength of 540 nm. When the absorbance of the wells containing only cells containing no cells was 0% and the absorbance of the wells without the sample was 100%, the concentration showing 50% absorbance was calculated as the IC ₅₀ value of each anticancer agent. The results are shown in Table 1.

Inhibitory Concentration on Cancer Cell Lines (IC ₅₀ , μg / mL) Concentration (μg / ml) A549 SK-OV-3 HCT15 SK-MEL2 Capecitabine 9.7 25.1 15.9 22.0 Example 3 0.85 0.72 0.80 0.65

As shown in Table 1, the compound of Formula 1 according to the present invention was confirmed to have good anticancer activity of about 0.6 ~ 0.9 ㎍ / ml for each cancer cell, this value is better than the control drug It can be seen that.

As described above, the compound of Formula 1 according to the present invention has a very good anticancer effect and less toxic, and thus an anticancer agent including it as an active ingredient may be used more effectively than a conventional anticancer agent.

Claims (5)

5′-deoxy-N-alkyloxycarbonyl-5-fluorocytosine-5′-thioester derivatives represented by Formula 1, pharmaceutically acceptable salts thereof, or solvates thereof: [Formula 1] Where R ₁ is hydrogen or C ₁ -C ₁₀ saturated or unsaturated hydrocarbon; R ₂ is hydrogen or an atomic group or easily removable protecting group that can be readily hydrolyzed under physiological conditions; R ₃ is a C ₂ -C ₇ saturated or unsaturated hydrocarbon. The compound of claim 1, wherein R ₂ in Formula 1 is hydrogen. a) combining the compound of Formula 2 with a thiol of Formula 3 in the presence of a solvent to prepare a compound of Formula 1b; And b) deprotecting the compound of Formula 1b to prepare a compound of Formula 1a Method for producing a compound of claim 1 comprising: [Formula 1a] [Formula 1b] [Formula 2] [Formula 3] R ₁ -SH Where R ₁ is hydrogen or C ₁ -C ₁₀ saturated or unsaturated hydrocarbon; R ₂ is hydrogen or an atomic group or easily removable protecting group that can be readily hydrolyzed under physiological conditions; R ₃ is a C ₂ -C ₇ saturated or unsaturated hydrocarbon. The method of claim 3, wherein the compound of Chemical Formula 2 of a) is prepared by protecting the compound of Chemical Formula 4 under solvent conditions. [Formula 4] Wherein R ₃ is a C ₂ -C ₇ saturated or unsaturated hydrocarbon. An anticancer composition comprising the compound of claim 1, a pharmaceutically acceptable salt thereof, or a solvate thereof as an active ingredient.