KR100631754B1 - N-alkyloxycarbonyl-5-fluorocytosine derivatives and preparation method thereof - Google Patents

Abstract

The present invention relates to a novel cytosine-based compound and a method for preparing the same, and more particularly, to N-alkyloxycarbonyl-5-fluorocytosine derivatives thereof having low toxicity and excellent anticancer effect, and a method for preparing the same. .

Cytosine, anticancer drugs,

Description

N-alkyloxycarbonyl-5-fluorocytosine derivatives and preparation method thereof {N-alkyloxycarbonyl-5-fluorocytosines and thier preparation}

The present invention relates to a novel cytosine compound and a method for producing the same, and more particularly, to an N-alkyloxycarbonyl-5-fluorocytosine derivative having low toxicity and excellent anticancer effect, and a method for producing the same.

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

Currently, local therapies such as surgery, radiation, and systemic therapies such as chemotherapy and immunotherapy are used to treat cancer. Among these, chemotherapy is widely used as an adjuvant drug for local therapy in the metastasis of various tumors and solid tumors that occur primarily in various organs, including gastrointestinal, liver, and lung, or as monotherapy.

Studies on cancer have been conducted by many scholars from several perspectives, including 5-Fluorouracil (5-FU), Methotrexate, Frutraful, Cisplatin, and others. In addition to the development of anticancer drugs, research is continuing on the development of new anticancer drugs. However, no formulations have yet been developed that can cure completely safely.

Recently, studies on the development of anticancer drugs using derivatives of 5-fluorouracil (5-FU), which are representative antipyretics of pyrimidine nucleosides, are being actively conducted. 5-FU, which is widely known as an anticancer agent, is used for the treatment of tumors by inhibiting the synthesis of pyrimidine nucleotides by activating metabolism in cells, but it has gastrointestinal toxicity, so its side effects are severe. Therefore, while many studies are being conducted to reduce this, some derivatives of 5-FU developed are activated in the intestinal wall after oral administration and cause diarrhea as a side effect.

Method for preparing N-alkyloxycarbonyl 5'-deoxy-5-fluorocytidine derivative represented by the following general formula (1) in European Patent No. 602454, Japanese Patent No. 94211891 and U.S. Patent No. 5472949 Is disclosed. However, these derivatives are known to be activated by enzymes in the liver rather than in the intestine, reducing their side effects, but they are not yet completely safe.

Wherein R ^a is a saturated or unsaturated hydrocarbon and R ^b represents hydrogen or an atomic group and easily removable protecting groups that are readily hydrolyzable under physiological conditions.

Accordingly, the present inventors conducted extensive research on a wide range of 5-FU series compounds and derivatives thereof to develop excellent anticancer agents. As a result, the present inventors have developed the N-alkyloxycarbonyl-5-fluorocytosine derivative of the present invention. As a result of measuring the effects, it was confirmed that the anticancer effect was excellent while the toxicity was low.

Accordingly, an object of the present invention is to provide a novel compound which is low in toxicity and excellent in anticancer effect.

Another object of the present invention is to provide a method for preparing the compound.

The present invention provides a compound of formula 2, a pharmaceutically acceptable salt or solvate thereof.

Wherein R ¹ is hydrogen, a C ₂ -C ₅ alkyl group or a C ₃ -C ₅ alkenyl group, R ² is hydrogen or an atomic group readily removable for physiological conditions and an easily removable protecting group, and R ³ is C ₂ -C ₇ alkyl group or alkenyl group.

The present invention also provides a compound of formula 3, a pharmaceutically acceptable salt or solvate thereof.

Wherein R ¹ is hydrogen, a C ₂ -C ₅ alkyl group, or a C ₃ -C ₅ alkenyl group, and R ² is hydrogen or an atomic group and easily removable protecting group that can be easily hydrolyzed under physiological conditions.

The present invention also provides a method for preparing a compound of formula 2b by reacting a compound of formula 3a with a compound of formula 4 below.

Wherein R ¹ is hydrogen, a C ₂ -C ₅ alkyl group, or a C ₃ -C ₅ alkenyl group, R ² is an atomic group and easily removable protecting group that can be readily hydrolyzed under physiological conditions, and R ³ is C _2- It represents an alkyl or alkenyl group of C _7.

The present invention also provides a method for preparing the compound of Formula 2a from the compound of Formula 2b.

In the above formula, R ¹ is hydrogen, a C ₂ -C ₅ alkyl group, or a C ₃ -C ₅ alkenyl group, and R ³ represents an alkyl group or an alkenyl group of C ₂ -C ₇ .

The present invention also provides a method for preparing a compound of formula 3a by reacting a compound of formula 5 with a compound of formula 6.

Wherein R ¹ is hydrogen, a C ₂ -C ₅ alkyl group, or a C ₃ -C ₅ alkenyl group, and R ² is an atomic group and easily removable protecting group that can be easily hydrolyzed under physiological conditions.

Hereinafter, a method for preparing the compound of Formula 2 of the present invention will be described in detail.

(Wherein R ¹ is hydrogen, a C ₂ -C ₅ alkyl group, or a C ₃ -C ₅ alkenyl group, R ² is an atomic group and an easily removable protecting group that can be easily hydrolyzed under physiological conditions, and R ³ is C ₂ − An alkyl group or an alkenyl group of C ₇ .

Referring to Scheme 1, compound (3a) is prepared by first adding a suitable additive from compound (5) and trimethylsilylated compound (6) in the presence of an acetonitrile solvent at -50 ° C to room temperature. The additive used at this time is preferably tin (IV) chloride, iodotrimethylsilane or chlorotrimethylsilane / sodium iodide.

Subsequently, compound (2b) is prepared by addition of a suitable base in the presence of a solvent from compound (4) wherein compound (3a) and R ³ are C ₂ -C ₇ alkyl or alkenyl at -50 ° C to room temperature. The solvent used in the reaction is preferably methylene chloride or pyridine, and the base is pyridine or triethylamine.

Next, the compound of formula (2a) is prepared by adding a suitable base from the formula (2b) in the presence of a solvent and stirring at room temperature.

In Scheme 1, Compound (5a) in which R ¹ is hydrogen in Compound (5), which was used as a reactant, was added from 0 ° C. to a suitable reagent in the presence of a suitable solvent and base from Compound (7a), as shown in Scheme 2 below. It can be obtained by stirring at room temperature. The solvent used is pyridine and methylene chloride, pyridine and triethylamine are suitable as bases, and acetic anhydride or acetyl chloride as reagents.

In addition, R ¹ is C ₂ -C ₅ in the compound (5) Compound (5b) which is an alkyl group and a C ₃ -C ₅ alkenyl group can be obtained by the same method as in Scheme 3 below.

Provided that R ¹ is a C ₂ -C ₅ alkyl group or a C ₃ -C ₅ alkenyl group

Referring to Scheme 3, p-toluenesulfonylchloride and Compound (10) were stirred in the presence of a solvent at room temperature to prepare Compound (9) therefrom, and copper iodide was removed in the presence of an ethyl ether solvent at -78 ° C to room temperature. In addition, C ₁ -C ₄ alkyllithium or C ₂ -C ₄ alkenyllithium is added dropwise thereto, followed by stirring to prepare compound (8) from compound (9). Subsequently, the compound (8) may be subjected to a deprotection reaction using an acid catalyst, followed by stirring at 0 ° C to room temperature in the presence of a suitable solvent and a base, to prepare an acetylated compound (5b).

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

Example 1 Methyl-5,6-dideoxy-2,3-O-isopropylidene-D-ribo-hexofuranose (Compound 8, R ^One = Ethyl)

60 ml of ethyl ether was added to 6.3 g of copper iodide in the presence of nitrogen, and methyllithium (1.6 M ethyl ether solution, 41 ml) was added dropwise thereto at -30 to 0 ° C. P-toluenesulfonyl made from 7.0 g of methyl-2,3-O-isopropylidene-D-ribofuranoside (10), 9.8 g of p-toluenesulfonyl chloride and 70 ml of pyridine at -78 ° C Substituted compound (9) was added dropwise. After stirring for 10 hours while gradually raising the temperature, 30 ml of ammonium chloride was added. The organic layer was separated, the aqueous layer was extracted several times with ethyl acetate, dried over anhydrous magnesium sulfate, filtered, distilled under reduced pressure, concentrated and purified by silica gel column chromatography to give 5.02 g (yield 76%) of the title compound.

¹ H NMR (CDCl ₃ , ppm) δ 4.94 (s, 1H), 4.81 (d, 1H), 4.56 (d, 1H), 4.40 (dd, 1H), 3,44 and 3.40 (s, 3H), 1,76 (m, 2H), 1,47 (s, 3H), 1.30 (s, 3H), 1.00 (t, 3H)

Example 2: 1,2,3-tri-O-acetyl-5,6-dideoxy-D-ribo-hexofuranose (Compound 5b, R ^One = Ethyl)

To 4.40 g of methyl-5,6-dideoxy-2,3-O-isopropylidene-D-ribo-hexofuranose (8), 40 ml of water and 1 ml of 1N aqueous hydrochloric acid solution were added and the mixture was heated at 80 to 120 ° C. After stirring for an hour, the solution was neutralized with 1N aqueous sodium hydroxide solution, and concentrated by distillation under reduced pressure. 20 ml of pyridine was added to dissolve it, 9 ml of acetic anhydride was added thereto, followed by stirring at room temperature for 9 hours. 50 ml of water was added thereto, extracted with 50 ml of chloroform three times, dried over anhydrous magnesium sulfate, and then filtered and distilled under reduced pressure. Then purified by silica gel column chromatography to give 5.49 g (92% yield) of the title compound.

¹ H NMR (CDCl ₃ , ppm) δ6.11 (d, 1H), 5.33 (dd, 1H), 5.09 (dd, 1H), 4.28 (m, 1H), 2.11 (s, 3H), 2.09 (s, 3H), 2.07 (s, 3H), 1.77 (m, 2H), 0.99 (t, 3H)

Example 3: Preparation of D-erythrofuranose-1,2,3-triacetate (5a)

To a solution of 4.0 g (85% purity) of D-erythrose (7a) and 25 ml of pyridine at 0 ° C., 9 ml of acetic anhydride was added dropwise, stirred at room temperature for 9 hours, concentrated by distillation under reduced pressure, and 50 ml of water. The mixture was extracted three times with 50 ml of chloroform, dried over anhydrous magnesium sulfate, filtered, distilled under reduced pressure, and concentrated to give 6.64 g (yield 95%) of the title compound.

¹ H NMR (CDCl ₃ , ppm) δ6.05 (d, 1H), 5.19 (dd, 1H), 5.12 (dd, 1H), 3.78 (dd, 1H), 3.63 (d, 1H), 2.14 (s, 3H), 2.10 (s, 3H), 2.06 (s, 3H)

Example 4-1: 1- (2 ', 3'-di-O-acetyl-β-D-erythrofuranosyl) -5-fluorocytosine

(Compound 3a, R ^One = H, R ² Preparation of = Ac)

Trimethylsilylated 5-fluorocytosine (6) prepared from 3.28 g of 5-fluorocytosine in 5.24 g of D-erythrofuranose 1,2,3-triacetate (5a) in the presence of nitrogen And 60 ml of acetonitrile were added. 3 ml of tin (IV) chloride was added dropwise thereto at -30 to 0 ° C, and stirred at room temperature for 4 to 8 hours. Then, an aqueous solution of sodium bicarbonate was added, extracted three times with 100 ml of ethyl acetate, and then dried and filtered over anhydrous magnesium sulfate. After distillation under reduced pressure, the mixture was purified by silica gel column chromatography to obtain 5.57 g (yield 83%) of the title compound.

¹ H NMR (CDCl ₃ , ppm) δ 8.27 (br.s, 2H), 7.25 (d, 1H), 5.67 (1H, d), 5.22 (1H, dd), 5.12 (1H, dd), 3.78 ( dd, 1H), 3.61 (d, 1H), 2.16 (s, 3H), 2.11 (s, 3H)

Example 4-2 1- (2 ', 3'-di-O-acetyl-5', 6'-dideoxy-β-D-ribo-hexofuranosyl) -5-fluoro-cytosine (compound 3a, R ^One Ethyl, R ² Preparation of = Ac)

1,2,3-tri-O-acetyl-5,6-dideoxy-D-ribo-hexofura of Example 2 instead of D-erythrofuranose 1,2,3, -triacetate (5a) The title compound was prepared in a similar manner as in Example 4-1, except that the nose 5b was used.

¹ H NMR (CDCl ₃ , ppm) δ 8.33 (br.s, 2H), 7.25 (d, 1H), 5.74 (d, 1H), 5.36 (dd, 1H), 5.10 (dd, 1H), 4.28 ( m, 1H), 2.13 (s, 3H), 2.11 (s, 3H), 2.08 (s, 3H), 1.73 (m, 2H), 0.98 (t, 3H)

Example 5-1: 1- (2 ', 3'-di-O-acetyl-5', 6'-dideoxy-β-D-ribo-hexofuranosyl) -5-fluoro-N ⁴ -(Propyloxycarbonyl) -cytosine (compound 2b, R ^One Ethyl, R ² = Ac, R ³ Propyl)

1- (2 ', 3'-di-O-acetyl-5', 6'-dideoxy-β-D-ribo-hexofuranosyl) -5-fluoro-cytosine prepared in Example 4-2 (3a) 20 ml of methylene chloride and 1.1 ml of pyridine were added to 2.33 g, and then 1.01 g of propyl chloroformate (4) was added dropwise at 0 ° C., stirred at room temperature for 30 minutes, followed by addition of aqueous sodium hydrogen carbonate solution and 50 ml of methylene chloride. The mixture was extracted three times with anhydrous magnesium sulfate, dried over anhydrous magnesium sulfate, filtered and concentrated under reduced pressure, and then purified by silica gel column chromatography to obtain 2.62 g (yield 90%) of the title compound.

¹ H NMR (CDCl ₃ , ppm) δ 12.00 (br.s, 1H), 7.61 (br.s, 1H), 6.07 (br.s, 1H), 5.28 (br.s, 1H), 5.27 (br .s, 1H), 4.37 (m, 3H), 2.15 (s, 3H), 2.08 (s, 3H), 1.78-1.66 (m, 2H), 1.33 (m, 2H), 1.01 (t, 3H), 0.94 (t, 3H)

Examples 5-2 to 5-7

In the same manner as in Example 5-1, except for using the compound (4) in which R ³ is ethyl, butyl, pentyl, hexyl, heptyl, and allyl instead of propylchloromate (4), Got it.

Example 5-2 1- (2 ', 3'-di-O-acetyl-5', 6'-dideoxy-β-D-ribo-hexofuranosyl) -5-fluoro-N ^4- ( Ethyloxycarbonyl) -cytosine (compound 2b, R ¹ = ethyl, R ² = Ac, R ³ = ethyl)

¹ H NMR (CDCl ₃ , ppm) δ 11.89 (br.s, 1H), 7.61 (br.s, 1H), 6.05 (d, 1H), 5.29 (dd, 1H), 5.13 (dd, 1H), 4.30 (m, 3H), 2.14 (s, 3H), 2.08 (s, 3H), 1.46 (m, 2H), 1.24 (t, 3H), 0.99 (t, 3H)

Example 5-3 1- (2 ', 3'-di-O-acetyl-5', 6'-dideoxy-β-D-ribo-hexofuranosyl) -5-fluoro-N ^4- ( Butyloxycarbonyl) -cytosine (Compound 2b, R ¹ = ethyl, R ² = Ac, R ³ = butyl)

¹ H NMR (CDCl ₃ , ppm) δ 11.85 (br.s, 1H), 7.61 (br.s, 1H), 6.04 (d, 1H), 5.30 (dd, 1H), 5.13 (dd, 1H), 4.33 (m, 3H), 2.13 (s, 3H), 2.10 (s, 3H), 1.57-1.33 (m, 6H), 1.02 (t, 3H), 0.98 (t, 3H)

Example 5-4 : 1- (2 ', 3'-di-O-acetyl-5', 6'-dideoxy-β-D-ribo-hexofuranosyl) -5-fluoro-N ^4- ( Pentyloxycarbonyl) -cytosine (Compound 2b, R ¹ = ethyl, R ² = Ac, R ³ = pentyl)

¹ H NMR (CDCl ₃ , ppm) δ 11.80 (br.s, 1H), 7.60 (br.s, 1H), 6.08 (d, 1H), 5.31 (dd, 1H), 5.12 (dd, 1H), 4.33 (m, 3H), 2.11 (s, 3H), 2.10 (s, 3H), 1.60-1.31 (m, 8H), 1.01 (t, 3H), 0.95 (t, 3H)

Example 5-5: 1- (2 ', 3'-di-O-acetyl-5', 6'-dideoxy-β-D-ribo-hexofuranosyl) -5-fluoro-N ^4- ( Hexyloxycarbonyl) -cytosine (compound 2b, R ¹ = ethyl, R ² = Ac, R ³ = hexyl)

¹ H NMR (CDCl ₃ , ppm) δ 11.93 (br.s, 1H), 7.61 (br.s, 1H), 6.11 (d, 1H), 5.30 (dd, 1H), 5.09 (dd, 1H), 4.35 (m, 3H), 2.13 (s, 3H), 2.09 (s, 3H), 1.63-1.30 (m, 10H), 1.00 (t, 3H), 0.94 (t, 3H)

Example 5-6 1- (2 ', 3'-di-O-acetyl-5', 6'-dideoxy-β-D-ribo-hexofuranosyl) -5-fluoro-N ^4- ( Heptyloxycarbonyl) -cytosine (compound 2b, R ¹ = ethyl, R ² = Ac, R ³ = heptyl)

¹ H NMR (CDCl ₃ , ppm) δ 12.00 (br.s, 1H), 7.58 (br.s, 1H), 6.11 (d, 1H), 5.30 (dd, 1H), 5.11 (dd, 1H), 4.32 (m, 3H), 2.13 (s, 3H), 2.08 (s, 3H), 1.65-1.29 (m, 12H), 1.01 (t, 3H), 0.96 (t, 3H)

Example 5-7 1- (2 ', 3'-di-O-acetyl-5', 6'-dideoxy-β-D-ribo-hexofuranosyl) -5-fluoro-N ^4- ( Allyloxycarbonyl) -cytosine (Compound 2b, R ¹ = ethyl, R ² = Ac, R ³ = allyl)

¹ H NMR (CDCl ₃ , ppm) δ 11.75 (br.s, 1H), 7.61 (br.s, 1H), 6.13 (d, 1H), 5.89 (m, 1H), 5.30 (dd, 1H), 5.23 (m, 4H), 4.75 (d, 2H), 4.33 (m, 1H), 2.13 (s, 3H), 2.10 (s, 3H), 1.46 (m, 2H), 0.98 (t, 3H)

Examples 5-8 to 5-14

1- (2 ', 3'-di-O-acetyl-5', 6'-dideoxy-β-D-ribo-hexofuranosyl) -5-fluoro-cytosine prepared in Example 4-2 (3a) 1- (2 ', 3'-di-O-acetyl-β-D-erythrofuranosyl) -5-fluorocytosine obtained in Example 4-1 instead (Compound 3, R ¹ = H, R ² = Ac), except that instead of propylchloromate (4) R ³ uses compound (4), each of which is ethyl, propyl, butyl, pentyl, hexyl, heptyl, allyl, In the same manner as in Example 5-1, the following compound was obtained.

Example 5-8: 1- (2 ', 3'-di-O-acetyl-β-D-erythrofuranosyl) -5-fluoro-N ^4- (ethyloxycarbonyl) -cytosine ( Compound 2b, R ¹ = H, R ² = Ac, R ³ = ethyl)

¹ H NMR (CDCl ₃ , ppm) δ 11.94 (br.s, 1H), 7.53 (br.s, 1H), 6.11 (d, 1H), 5.41 (dd, 1H), 5.12 (dd, 1H), 4.20 (q, 2H), 3.93 (dd, 1H), 3.85 (d, 1H), 2.16 (s, 3H), 2.13 (s, 3H), 1.28 (t, 3H)

Example 5-9: 1- (2 ', 3'-di-O-acetyl-β-D-erythrofuranosyl) -5-fluoro-N ^4- (propyloxycarbonyl) -cytosine ( Compound 2b, R ¹ = H, R ² = Ac, R ³ = propyl)

¹ H NMR (CDCl ₃ , ppm) δ 11.90 (br.s, 1H), 7.55 (br.s, 1H), 6.10 (d, 1H), 5.44 (dd, 1H), 5.12 (dd, 1H), 4.16 (t, 2H), 3.94 (dd, 1H), 3.83 (d, 1H), 2.16 (s, 3H), 2.14 (s, 3H), 1.61 (m, 2H), 0.99 (t, 3H)

Example 5-10: 1- (2 ', 3'-di-O-acetyl-β-D-erythrofuranosyl) -5-fluoro-N ^4- (butyloxycarbonyl) -cytosine ( Compound 2b, R ¹ = H, R ² = Ac, R ³ = butyl)

¹ H NMR (CDCl ₃ , ppm) δ 11.90 (br.s, 1H), 7.55 (br.s, 1H), 6.16 (d, 1H), 5.48 (dd, 1H), 5.15 (dd, 1H), 4.16 (t, 2H), 3.92 (dd, 1H), 3.81 (d, 1H), 2.16 (s, 3H), 2.12 (s, 3H), 1.57-1.33 (m, 4H), 0.98 (t, 3H)

Example 5-11 : 1- (2 ', 3'-di-O-acetyl-β-D-erythrofuranosyl) -5-fluoro-N ^4- (pentyloxycarbonyl) -cytosine ( Compound 2b, R ¹ = H, R ² = Ac, R ³ = pentyl)

¹ H NMR (CDCl ₃ , ppm) δ 11.69 (br.s, 1H), 7.54 (br.s, 1H), 6.14 (d, 1H), 5.46 (dd, 1H), 5.13 (dd, 1H), 4.11 (t, 2H), 3.92 (dd, 1H), 3.80 (d, 1H), 2.18 (s, 3H), 2.13 (s, 3H), 1.58-1.28 (m, 6H), 0.99 (t, 3H)

Example 5-12 : 1- (2 ', 3'-di-O-acetyl-β-D-erythrofuranosyl) -5-fluoro-N ^4- (hexyloxycarbonyl) -cytosine (Compound 2b, R ¹ = H, R ² = Ac, R ³ = hexyl)

¹ H NMR (CDCl ₃ , ppm) δ 11.88 (br.s, 1H), 7.54 (br.s, 1H), 6.16 (d, 1H), 5.45 (dd, 1H), 5.11 (dd, 1H), 4.11 (t, 2H), 3.93 (dd, 1H), 3.80 (d, 1H), 2.16 (s, 3H), 2.12 (s, 3H), 1.60-1.28 (m, 8H), 0.98 (t, 3H)

Example 5-13: 1- (2 ', 3'-Di-O-acetyl-β-D-erythrofuranosyl) -5-fluoro-N ^4- (heptyloxycarbonyl) -cytosine ( Compound 2b, R ¹ = H, R ² = Ac, R ³ = heptyl)

¹ H NMR (CDCl ₃ , ppm) δ 11.80 (br.s, 1H), 7.51 (br.s, 1H), 6.11 (d, 1H), 5.43 (dd, 1H), 5.09 (dd, 1H), 4.08 (t, 2H), 3.90 (dd, 1H), 3.77 (d, 1H), 2.14 (s, 3H), 2.10 (s, 3H), 1.60-1.24 (m, 10H), 0.98 (t, 3H)

Example 5-14: 1- (2 ', 3'-Di-O-acetyl-β-D-erythrofuranosyl) -5-fluoro-N ^4- (allyloxycarbonyl) -cytosine ( Compound 2b, R ¹ = H, R ² = Ac, R ³ = allyl)

¹ H NMR (CDCl ₃ , ppm) δ 11.66 (br.s, 1H), 7.52 (br.s, 1H), 6.12 (d, 1H), 5.88 (m, 1H), 5.46 (dd, 1H), 5.20 (m, 3H), 4.80 (d, 2H), 3.94 (dd, 1H), 3.79 (d, 1H), 2.15 (s, 3H), 2.09 (s, 3H)

Example 6-1: 1-β-D-erythrofuranosyl-5-fluoro-N ⁴ -(Pentyloxycarbonyl) -cytosine (compound 2a, R ^One = H, R ³ Pentyl)

1- (2 ', 3'-di-O-acetyl-β-D-erythrofuranosyl) -5-fluoro-N ^4- (pentyloxycarbonyl) -cytosine obtained in Examples 5-11 (2b) After dissolving 50 ml of methanol in 6.08 g, 50 ml of 1N sodium methoxide was added thereto, stirred for 1 hour, neutralized with 1N hydrochloric acid, and concentrated by distillation under reduced pressure. 30 ml of water was added thereto, followed by extraction several times with a methylene chloride / methanol (95/5) solution, followed by dry filtration with anhydrous magnesium sulfate, distillation under reduced pressure, concentration, and recrystallization with ethyl acetate to obtain 3.78 g (yield of the title compound). 85%) was obtained.

¹ H NMR (CD ₃ OD, ppm) δ 7.88 (d, 1H), 5.85 (br.s, 1H), 4.15 (t, 2H), 4.05 (dd, 1H), 3.98 (dd, 1H), 3.93 (dd, 1H), 3.83 (d, 1H), 1.58-1.28 (m, 6H), 0.99 (t, 3H)

Examples 6-2 to 6-7

1- (2 ', 3'-di-O-acetyl-β-D-erythrofuranosyl) -5-fluoro-N ^4- (pentyloxycarbonyl) -cytosine obtained in Examples 5-11 The following compound (2a) was prepared in the same manner as in Example 6a, except for using the compounds of Examples 5-8 to 5-10 and 5-12 to 5-14 instead of (2b).

Example 6-2 1-β-D-erythrofuranosyl-5-fluoro-N ^4- (ethyloxycarbonyl) -cytosine (Compound 2a, R ¹ = H, R ³ = ethyl)

¹ H NMR (CD ₃ OD, ppm) δ 7.88 (d, 1H), 5.84 (br.s, 1H), 4.20 (q, 2H), 4.03 (dd, 1H), 3.99 (dd, 1H), 3.92 (dd, 1H), 3.80 (d, 1H), 1.30 (t, 3H)

Example 6-3 1-β-D-erythrofuranosyl-5-fluoro-N ^4- (propyloxycarbonyl) -cytosine (Compound 2a, R ¹ = H, R ³ = propyl)

¹ H NMR (CD ₃ OD, ppm) δ 7.85 (d, 1H), 5.82 (br.s, 1H), 4.15 (t, 2H), 4.00 (dd, 1H), 3.98 (dd, 1H), 3.92 (dd, 1H), 3.77 (d, 1H), 1.61 (m, 2H), 0.99 (t, 3H)

Example 6-4 : 1-β-D-erythrofuranosyl-5-fluoro-N ^4- (butyloxycarbonyl) -cytosine (Compound 2a, R ¹ = H, R ³ = butyl)

¹ H NMR (CD ₃ OD, ppm) δ 7.85 (d, 1H), 5.85 (br.s, 1H), 4.14 (t, 2H), 3.99 (dd, 1H), 3.95 (dd, 1H), 3.88 (dd, 1H), 3.79 (d, 1H), 1.57-1.33 (m, 4H), 0.98 (t, 3H)

Example 6-5 : 1-β-D-erythrofuranosyl-5-fluoro-N ^4- (hexyloxycarbonyl) -cytosine (Compound 2a, R ¹ = H, R ³ = hexyl)

¹ H NMR (CD ₃ OD, ppm) δ 7.83 (d, 1H), 5.81 (br.s, 1H), 4.13 (t, 2H), 4.01 (dd, 1H), 3.95 (dd, 1H), 3.86 (dd, 1H), 3.79 (d, 1H), 1.60-1.28 (m, 8H), 0.98 (t, 3H)

Example 6-6 1-β-D-erythrofuranosyl-5-fluoro-N ^4- (heptyloxycarbonyl) -cytosine (Compound 2a, R ¹ = H, R ³ = heptyl)

¹ H NMR (CD ₃ OD, ppm) δ 7.83 (d, 1H), 5.80 (br.s, 1H), 4.13 (t, 2H), 4.02 (dd, 1H), 3.96 (dd, 1H), 3.88 (dd, 1H), 3.80 (d, 1H), 1.62-1.25 (m, 10H), 0.97 (t, 3H)

Example 6-7 1-β-D-erythrofuranosyl-5-fluoro-N ^4- (allyloxycarbonyl) -cytosine

(Compound 2a, R ¹ = H, R ³ = Allyl)

¹ H NMR (CD ₃ OD, ppm) δ 7.85 (d, 1H), 5.88 (m, 2H), 5.24 (m, 2H), 4.75 (d, 2H), 4.01 (dd, 1H), 3.95 (dd , 1H), 3.88 (d, 1H), 3.78 (d, 1H)

Examples 6-8 to 6-14

1- (2 ', 3'-di-O-acetyl-β-D-erythrofuranosyl) -5-fluoro-N ^4- (pentyloxycarbonyl) -cytosine obtained in Examples 5-11 The following compounds were prepared in the same manner as in Example 6a, except that the compounds of Examples 5-1 to 5-7 were used instead of (2b).

Example 6-8: 1- (5 ', 6'-dideoxy-β-D-hexofuranosyl) -5-fluoro-N ^4- (ethyloxycarbonyl) -cytosine (Compound 2a, R ¹ = Ethyl, R ³ = ethyl)

¹ H NMR (CD ₃ OD, ppm) δ 7.88 (d, 1H), 5.83 (br.s, 1H), 4.29 (dd, 1H), 4.17 (q, 2H), 3.88 (dd, 1H), 3.66 (dd, 1H), 1.49 (m, 2H), 1.30 (t, 3H), 0.99 (t, 3H)

Example 6-9 1- (5 ′, 6′-dideoxy-β-D-hexofuranosyl) -5-fluoro-N ^4- (propyloxycarbonyl) -cytosine (Compound 2a, R ¹ = Ethyl, R ³ = propyl)

¹ H NMR (CD ₃ OD, ppm) δ 7.88 (d, 1H), 5.83 (br.s, 1H), 4.28 (dd, 1H), 4.15 (t, 2H), 3.82 (dd, 1H), 3.65 (dd, 1H), 1.61-1.46 (m, 4H), 1.02 (t, 3H), 0.96 (t, 3H)

Example 6-10: 1- (5 ', 6'-dideoxy-β-D-hexofuranosyl) -5-fluoro-N ^4- (butyloxycarbonyl) -cytosine (Compound 2a, R ¹ = Ethyl, R ³ = butyl)

¹ H NMR (CD ₃ OD, ppm) δ 7.85 (d, 1H), 5.82 (br.s, 1H), 4.26 (dd, 1H), 4.13 (t, 2H), 3.80 (dd, 1H), 3.65 (dd, 1H), 1.61-1.33 (m, 6H), 1.02 (t, 3H), 0.97 (t, 3H)

Example 6-11 1- (5 ', 6'-dideoxy-β-D-hexofuranosyl) -5-fluoro-N ^4- (pentyloxycarbonyl) -cytosine (Compound 2a, R ¹ = Ethyl, R ³ = pentyl)

¹ H NMR (CD ₃ OD, ppm) δ 7.86 (d, 1H), 5.82 (br.s, 1H), 4.28 (dd, 1H), 4.12 (t, 2H), 3.81 (dd, 1H), 3.61 (dd, 1H), 1.62-1.29 (m, 8H), 1.04 (t, 3H), 0.97 (t, 3H)

Example 6-12 1- (5 ', 6'-dideoxy-β-D-hexofuranosyl) -5-fluoro-N ^4- (hexyloxycarbonyl) -cytosine (Compound 2a, R ¹ = ethyl, R ³ = hexyl)

¹ H NMR (CD ₃ OD, ppm) δ 7.89 (d, 1H), 5.80 (br.s, 1H), 4.25 (dd, 1H), 4.13 (t, 2H), 3.83 (dd, 1H), 3.58 (dd, 1H), 1.61-1.29 (m, 10H), 1.02 (t, 3H), 0.97 (t, 3H)

Example 6-13 1- (5 ', 6'-dideoxy-β-D-hexofuranosyl) -5-fluoro-N ^4- (heptyloxycarbonyl) -cytosine (Compound 2a, R ¹ = Ethyl, R ³ = heptyl)

¹ H NMR (CD ₃ OD, ppm) δ 7.81 (d, 1H), 5.83 (br.s, 1H), 4.23 (dd, 1H), 4.13 (t, 2H), 3.85 (dd, 1H), 3.55 (dd, 1H), 1.61-1.28 (m, 12H), 1.01 (t, 3H), 0.97 (t, 3H)

Example 6-14 1- (5 ', 6'-dideoxy-β-D-hexofuranosyl) -5-fluoro-N ^4- (allyloxycarbonyl) -cytosine (Compound 2a, R ¹ = Ethyl, R ³ = allyl)

¹ H NMR (CD ₃ OD, ppm) δ 7.88 (d, 1H), 5.85 (m, 2H), 5.25 (m, 2H), 4.78 (d, 2H), 4.28 (dd, 1H), 3.58 (dd , 1H), 3.61 (dd, 1H), 1.46 (m, 2H), 0.98 (t, 3H)

Example 7 Drug Evaluation of Compound (2)

Capecitabine (5'-deoxy-5-fluoro) showing the best result among 5'-deoxy-N-alkyloxycarbonyl-5-fluorocytidine derivatives disclosed in EP 602454. N-alkyloxycarbonyl-5, a compound of Formula 2, according to the present invention with rho-N ^4- (pentyloxycarbonyl) cytidine (Compound 1 wherein R ^a = pentyl, R ^b = H) as a control agent In order to measure the anticancer activity of fluorocytosine derivatives, cytotoxicity against human cancer cells was measured, where human cancer cells were A549 (lung cancer) HCT15 (colon cancer) SK-OV-3 (ovarian cancer) SK- Comparison was made using MEL-2 (skin cancer).

First, the cancer cells were cultured in a 37 ° C., 5% CO ₂ thermohygrostat, and 10% fetal calf serum was added using RPMI as a basal medium. To measure the cytotoxic activity, the cancer cells in the logarithmic phase were divided into 2-5 x 10 ⁴ cells per well of a 96-well plate and incubated for 24 hours, followed by capecitabine and 1-β-D-eryth. Thrurofuranosyl-5-fluoro-N ^4- (pentyloxycarbonyl) -cytosine (Compound 2a, R ¹ = H, R ³ = pentyl), 1- (5 ', 6'-dideoxy-β -D-ribo-hexofuranosyl) -5-fluoro-N ^4- (pentyloxycarbonyl) -cytosine (Compound 2a, R ¹ = ethanol, R ² = H, R ³ = pentyl) One sample solution was added and incubated for 72 hours. To each well of the cultured plate, 20 μl of MTT reaction solution dissolved in 5 mg / ml concentration in physiological saline was added and incubated for 4 hours, and the formed formazan crystal was 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 , ㎍ / ml) Concentration (㎍ / ml) A549 SK-OV-3 HCT15 SK-MEL2 Capecitabine 8.791 23.775 13.868 23.067 1-β-D-erythrofuranosyl-5-fluoro-N ^4- (pentyloxycarbonyl) -cytosine 1.430 2.526 4.122 2.889 1- (5 ', 6'-dideoxy-β-D-ribo-hexofuranosyl) -5-fluoro-N ^4- (pentyloxycarbonyl) -cytosine ( 2.292 5.819 3.142 6.222

As shown in Table 1, the compound of Formula 2 according to the present invention confirmed a strong anticancer activity against each cancer cell, it can be seen that this value represents a much better anticancer activity than the reference drug.

As described above, the N-alkyloxycarbonyl-5-fluorocytosine derivatives according to the present invention can be used as a very useful anticancer agent.

Claims (5)

A compound of Formula 2, a pharmaceutically acceptable salt or solvate thereof:

Wherein R ¹ is hydrogen, a C ₂ -C ₅ alkyl group, or a C ₃ -C ₅ alkenyl group, R ² is a hydrogen, an acetyl group or a 2,3-O-isopropylidene group, and R ³ is a C ₂ An alkyl group or an alkenyl group of -C ₇ . A compound of Formula 3, a pharmaceutically acceptable salt or solvate thereof:

Wherein R ¹ is hydrogen, a C ₂ -C ₅ alkyl group, or a C ₃ -C ₅ alkenyl group, and R ² is a hydrogen, an acetyl group or a 2,3-O-isopropylidene group. A method of preparing a compound of Formula 2b by reacting a compound of Formula 3a with a compound of Formula 4:

Wherein R ¹ is hydrogen, a C ₂ -C ₅ alkyl group, or a C ₃ -C ₅ alkenyl group, R ² is an acetyl group or a 2,3-O-isopropylidene group, and R ³ is C ₂ -C The alkyl group or alkenyl group of ₇ is shown. A process for preparing a compound of formula 2a from a compound of formula 2b using a base:

Wherein R ¹ is hydrogen, a C ₂ -C ₅ alkyl group, or a C ₃ -C ₅ alkenyl group, R ² is an acetyl group or a 2,3-O-isopropylidene group, and R ³ is C ₂ -C The alkyl group or alkenyl group of ₇ is shown. A process for preparing a compound of formula 3a by reacting a compound of formula 5 with a compound of formula 6

In the above formula, R ¹ is hydrogen, a C ₂ -C ₅ alkyl group, or a C ₃ -C ₅ alkenyl group, and R ² is an acetyl group or a 2,3-O-isopropylidene group.