JPH04282397A - Ethylenediaminediacetic acid compound and production thereof - Google Patents

Abstract

PURPOSE:To obtain the title new compound useful as a treating agent for gallstone, having gallstone dissolving action by reacting a mixed acid anhydride of usodeoxycholic acid with a protected ethylenediamine, amidating, deprotecting and reacting the resulting substance with a halogenated acetic acid. CONSTITUTION:A mixed acid anhydride of usodeoxycholic acid shown by formula I (R<1> is 1-4C alkyl) is condensed with an ethylenediamine shown by formula II (R<2> is H, benzyloxycarbonyl or t-butyloxycarbonyl) to give a ursodeoxycholylethylenediamine compound shown by formula III, this amide compound is catalytically reduced or hydrolyzed with an acid, deprotected and reacted with a halogenated acetic acid shown by the formula X-CH2COOH (X is chlorine, bromine or iodine) in the presence of a base to give the objective ursodeoxycholylethylenediaminediacetic acid (e.g. ursodeoxycholylethylenediamine-N,N-diacetic acid) shown by formula IV.

Description

[Detailed description of the invention]

0001

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to ethylenediaminediacetic acid compounds, and more particularly to ursodeoxycholylethylenediaminediacetic acid compounds having gallstone dissolving activity and a method for producing the same.

0002

BACKGROUND OF THE INVENTION Ursodeoxycholic acid is known as a drug commonly used as a gallstone treatment agent.

Furthermore, JP-A-60-161996 discloses that a compound containing amide bonds of ursodeoxycholic acid or chenodeoxycholic acid and aspartic acid, glutamic acid, serine or carboxymethylglycine is used to treat cholesterol-based gallstones with calcified outer shells. It has been reported that it has the effect of dissolving.

0004

[Problems to be Solved by the Invention] However, ursodeoxycholic acid and chenodeoxycholic acid are only effective against pure cholesterol stones, and are effective against other cholesterol-based gallstones, such as cholesterol-containing calcium-containing cholesterol stones or cholesterol-mixed stones. , Furthermore, for bilirubin calcium stone or calcium carbonate stone, etc.
Its dissolving effect has been questioned.

On the other hand, the compound described in JP-A No. 60-161996 has a biochemical effect on N-ursodeoxycholylu-N-carboxymethylglycine, which is said to have the highest dissolving effect on cholesterol gallstones with calcified outer shell. The amount is only about 2 to 3 times that of glycochenodeoxycholic acid, which is a typical compound present in the world.

As a result of intensive research into bile acid derivatives, the present inventors found that ursodeoxycholylethylenediamine diacetic acid compound strongly dissolves calcium-containing gallstones, especially calcium carbonate-containing gallstones in bile. reached.

[0007]

[Means for Solving the Problems] According to the present invention, the following structural formula

[0008]

[Chemical formula 1]

The present invention provides a ursodeoxycholylethylenediaminediacetic acid compound (hereinafter referred to as "Compound I") represented by the following formula, a physiologically acceptable salt thereof, and a method for producing the same.

In the present invention, physiologically acceptable salts refer to mono-, di- or trial-alkali salts, mono- or di-mineral salts or triammonium salts. Examples of the alkali salt include sodium salt or potassium salt, and examples of the mineral acid salt include hydrochloride, sulfate, or nitrate.

Compound [I] can be produced through the following reaction steps.

First, the following general formula

[0013]

[Case 2]

(In the formula, R1 represents a linear or branched alkyl group having 1 to 4 carbon atoms.) A mixed acid anhydride (hereinafter referred to as "compound [II]") represented by the following general formula

[0015]

[Chemical formula 3]

(In the formula, R2 represents a hydrogen atom, a benzyloxycarbonyl group, a t-butyloxycarbonyl group, or a trityl group.) (hereinafter referred to as "compound [III]") is condensed with The resulting general formula

[0017]

[C4]

A method of catalytic reduction or hydrolysis with an acid (hereinafter referred to as the "first method") of the amide compound represented by the formula (wherein R2 has the same meaning as above) (hereinafter referred to as "compound [IV]") ), or by a method of directly condensing compound [II] and ethylenediamine (hereinafter referred to as "second method"), the formula

[0019]

[C5]

An ethylenediamine compound (hereinafter referred to as "compound [V]") represented by the following formula is produced.

[0021] Compound [II] and compound [
Regarding the reaction ratio of compound [III], the molar amount of compound [III] is 0.5 to 2 times that of compound [II]. The reaction solvent is
Tetrahydrofuran, dioxane, methanol, chloroform, water, or a mixture of two or more thereof is suitable. The reaction temperature is -30 to 20°C, preferably -10 to 10°C, and the reaction time is 10 minutes to 20°C.
The period is 48 hours, preferably about 1 to 12 hours.

The starting compound [II] can be produced by reacting ursodeoxycholic acid with an alkyl chloroformate in the presence of an acid acceptor.

The reaction ratio is approximately equimolar amount of alkyl chloroformate to ursodeoxycholic acid. Examples of the alkyl chloroformate used include ethyl chloroformate and isobutyl chloroformate. Examples of acid acceptors include triethylamine, tributylamine, and N-methylmorpholine. Dioxane or tetrahydrofuran is suitable as the reaction solvent. The reaction temperature is -
The temperature is 20 to 10°C, and the reaction time is 1 minute to 3 hours. This reaction proceeds almost quantitatively, and the obtained compound [
II] is unstable, so the reaction mixture is used in the next step without isolation.

Compound [III
] is a general amino protecting group introduction method (
Fundamentals and Experiments of Peptide Synthesis, pages 17-39, 1982
(Published by Maruzen Co., Ltd. on January 20, 2016).

[0025] For the catalytic reduction in the first method, a commonly used method can be adopted. Examples of acids used in hydrolysis include hydrochloric acid, acetic acid, and a mixture thereof.

The reaction solvent used in the second method is suitably tetrahydrofuran, dioxane, methanol, chloroform, water, or a mixture of two or more thereof. The reaction temperature is -30 to 20°C, preferably -10 to 1
The temperature should be within the range of 0°C, and the reaction time should be about 10 minutes to 48 hours, preferably about 1 to 12 hours.

[0027] Compound [V] can be produced by the first method or the second method described above, but considering the purity of the triamine compound [V] obtained and the treatment in the next step, the first method is preferable. It is preferable to adopt it.

Next, compound [V] is expressed by the following general formula:

0029
]

[C6]

(In the formula, X represents chlorine, bromine or iodine.) Compound [I] is obtained by reacting it with a halogenated acetic acid represented by (hereinafter referred to as "compound [VI]") in the presence of a base. can get.

The reaction ratio is such that the molar amount of compound [VI] is 3 to 10 times that of compound [V]. Water is suitable as the reaction solvent. Examples of the base used include sodium hydrogen carbonate, sodium carbonate, and potassium hydroxide. The reaction temperature is 30-95°C, preferably 40-60°C
The reaction time is 1 to 48 hours, preferably 5 to 24 hours.

Compound [I] can be separated in the form of a free acid by adjusting the pH of the basic solution at the end of the reaction to about 2.5 using a mineral acid such as hydrochloric acid or sulfuric acid. The resulting free acid is purified by column chromatography, if necessary.

The physiologically acceptable salt of compound [I] is prepared by adding an excess amount of aqueous ammonia, 1, 2 or 3 equivalents of alkali or 1 or 2 equivalents of mineral acid to compound [I] or an aqueous solution thereof. It can be produced by adding and then drying this to dryness under reduced pressure. Examples of the alkali used include sodium hydrogen carbonate, potassium carbonate, sodium hydroxide, and potassium hydroxide, and examples of the mineral acid include hydrochloric acid, sulfuric acid, and nitric acid.

[0034]

[Action] The dissolving action of the ethylenediaminediacetic acid compound on calcium carbonate-containing gallstones will be explained in detail below. The solubilizing effect was evaluated by the ability to dissolve calcium carbonate (CaCO3) in bile.

[0035] In the test, an excess amount of calcium carbonate was added to 2 ml of the artificial bile model, and after incubation at 37°C for 18 hours under a sealed stopper, the mixture was centrifuged at 3000 rpm, and the amount of calcium carbonate dissolved in the resulting supernatant was determined. was measured using an atomic absorption spectrometer. Diethylenetriaminetriacetic acid compound was dissolved in 0.01M phosphate buffer (pH 7.4 and 8.3) to prepare a concentration of 16mM, lecithin of 8mM, and cholesterol of 4mM.

[0036] Ethylenediaminediacetic acid compound is converted into glycochenodeoxycholic acid or N-ursodeoxycholyl-N
- The same treatment as above except that carboxymethylglycine was used, and the ability of these two compounds to dissolve calcium carbonate was tested for comparison.

The results are shown in the table below. Note that the numerical values in parentheses in the same table are those described in Japanese Patent Application Laid-open No. 161996/1983, and are also listed for reference.

[0038]

[Table 1]

As is clear from the above table, the ethylenediaminediacetic acid compound of the present invention has far superior ability to dissolve calcium carbonate than glycochenodeoxycholic acid and N-ursodeoxycholyl-N-carboxymethylglycine. It is recognized that there are

The present invention will be further explained by referring to reference examples and examples.

[0041]

[Reference Example 1] (Compound [II]) 11.8 g (30.1 mmol) of ursodeoxycholic acid, 50 ml of tetrahydrofuran, and 4.30 ml (30.1 mmol) of triethylamine.
8 mmol), 4.00 ml (30.8 mmol) of isobutyl chloroformate was added dropwise at -4 to -1°C,
The mixture was stirred at −2 to 1° C. for 2 hours to obtain a reaction mixture containing 14.8 g of isobutyl ursodeoxycholyl carbonate (yield quantitative).

[0042]

[Reference Example 2] (Compound [III]) Ethylenediamine 1
33 ml (1.99 mol) was dissolved in 200 ml of chloroform, and to this was added 111.7 g (0.99 mol) of trityl chloride.
A solution of 401 mol) dissolved in 860 ml of chloroform was added dropwise under ice cooling, and the mixture was stirred overnight at room temperature. This reaction solution was washed with water, dried over anhydrous sodium sulfate, and then the solvent was distilled off under reduced pressure. The obtained residue was subjected to alumina column chromatography using a chloroform-methanol mixture (volume ratio 5:1) as a developing solution to obtain 108.9 g of oily N-tritylethylenediamine. The yield based on trityl chloride was 89.9%.

[0043]

[Example 1] (Compound [IV]) Tetrahydrofuran 4
10.1 g of N-tritylethylenediamine (3
14.8 g (30.1 mmol) of isobutylursodeoxycholyl carbonate was added at -6 to 0°C to a solution in which 3.3 mmol) was dissolved, and the mixture was further stirred at -8 to -3°C for 1 hour. The solvent of the resulting reaction solution was distilled off under reduced pressure, and the residue was dissolved in a chloroform-methanol mixture (volume ratio 50:1).
) was subjected to alumina column chromatography using N-trityl-N'-ursodeoxycholylethylenediamine as a developing solution to obtain 14.1 g of a glassy substance (yield: 69.5
%) was obtained.

(Compound [V]) 2.89 g of N-trityl-N'-ursodeoxycholylethylenediamine (4.
Dissolve 28 mmol) in 15 ml of acetic acid, and add 5 ml of water to this.
1 was added and stirred at 36-40°C for 1.2 hours. After cooling, the precipitate was filtered, and 140 ml of water was added to the filtrate to give a concentration of 17% (W
/V) The pH was adjusted to 11 with an aqueous sodium hydroxide solution, and then this was extracted with n-butanol. The n-butanol layer was washed with water, dried over anhydrous sodium sulfate, and then dried under reduced pressure to obtain 1.83 g (yield: 98.5%) of a colorless glassy substance of N'-ursodeoxycholylethylenediamine.

(Compound [I]) 2.43 g of bromoacetic acid (
17.5 mmol) was dissolved in 15 ml of water, and the solution was adjusted to pH 7.2 using an 8% aqueous sodium carbonate solution. This prepared solution was added at 50°C to 15 ml of water containing 1.45 g (3.35 mmol) of N'-ursodeoxycholylethylenediamine. Then, while stirring at 50°C, an 8% aqueous sodium carbonate solution was added dropwise to this mixture, first adjusting the pH to 7.5 to 8.5 over 1.5 hours, and then adjusting the final pH to 8 over 12 hours. It was adjusted to .0 to 8.5. This reaction solution was cooled and adjusted to pH 2.5 with 1N hydrochloric acid.
and extracted using n-butanol. After washing the extract with saturated saline and drying with anhydrous sodium sulfate,
The solvent was distilled off. The residue was subjected to siligal column chromatography using a mixture of ethanol and 28% ammonia water (volume ratio 9:1) as a developing solution, and the eluate was dried under reduced pressure. Add 20 ml of water to the resulting residue, and adjust the pH with 1N hydrochloric acid.
It was set at 2.5. The resulting precipitate was collected by filtration, dried under reduced pressure,
N'-Ursodeoxycholylethylenediamine-N,N
-0.87 g (yield 49.5%) of colorless powder of diacetic acid was obtained.

[0046]

[0047]

[Example 2] (Compound [V]) 20 ml of dioxane and isobutyl ursodeoxycholyl carbonate6.
The mixed solution was added dropwise to 2 g (12.7 mmol) and 17.2 ml (233.3 mmol) of ethylenediamine little by little at 5 to 10°C, and stirred at the same temperature for 3 hours. This reaction solution was poured into water and extracted with ethyl acetate. This ethyl acetate layer was washed with saturated brine, dried over anhydrous sodium sulfate, and then the solvent was distilled off to obtain 3.3 g of a viscous substance of N'-ursodeoxycholylethylenediamine. The purity of this material was 68%.

(Compound [I]) Using 3.3 g of the obtained viscous substance of N'-ursodeoxycholylethylenediamine, the process was carried out in almost the same manner as in Example 1, except that the conditions were slightly changed. -Ursodeoxycholylethylenediamine-N,N-diacetic acid colorless powder 0.68 (yield 17.2)
%) was obtained. Melting point, IR spectrum and 1H of this powder
-NMR spectrum was consistent with that described in Example 1.

[0049]

[Example 3] (Physiologically acceptable salt of compound [I]) The eluate obtained by silica gel column chromatography in Example 1 was concentrated, an appropriate amount of toluene was added, and the water in the concentrate was was removed azeotropically and then dried. Next, the residue was dissolved in a small amount of methanol, and an appropriate amount of ethyl acetate was added thereto to crystallize it, resulting in 1.05 g of N'-ursodeoxycholylethylenediamine-N,N-diacetic acid diammonium salt (yield: 56.4 %) was obtained.

[0050]

[0051]

[Effect] The ethylenediaminediacetic acid compound and its physiologically acceptable salts of the present invention can be used as a dissolving agent for various calcium-containing gallstones.

Claims (2)

[Claims] 1. An ethylenediaminediacetic acid compound represented by the formula: [Image Omitted] and a physiologically acceptable salt thereof. Claim 2: A mixed acid anhydride represented by the general formula [Formula 2] (wherein R1 represents a linear or branched alkyl group having 1 to 4 carbon atoms) and a mixed acid anhydride represented by the general formula [Formula 3] (Formula (wherein, R2 represents a hydrogen atom, a benzyloxycarbonyl group, a t-butyloxycarbonyl group, or a trityl group).
] (In the formula, R2 has the same meaning as above.) By catalytic reduction or acid hydrolysis of the amide compound represented by the formula, or by directly condensing the above mixed acid anhydride and ethylenediamine, the formula [Chemical formula 5] is obtained. A claim characterized in that an ethylenediamine compound represented by the formula is prepared and then reacted with a halogenated acetic acid represented by the general formula [Chemical formula 6] (wherein X represents chlorine, bromine or iodine) in the presence of a base. Item 2. A method for producing an ethylenediaminediacetic acid compound and a physiologically acceptable salt thereof according to item 1.