JPS5910354B2 - Method for producing N-leucyl agmatine compound - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a method for producing a novel N-leucyl agmatine compound, and more specifically, it is represented by the general formula (wherein R represents a lower alkyl group).

) The present invention relates to a method for producing an N-leucyl agmatine compound represented by: The first invention of the present invention provides N-leucyl agmatine represented by the general formula (wherein R represents a lower alkyl group).

) is reacted with the compound represented by the general formula (wherein R has the same meaning as above).

) or a salt thereof. The second aspect of the present invention is to catalytically reduce and hydrolyze a compound represented by the general formula to N-leucyl agmatine represented by the formula, and the resulting formula (wherein R represents a lower alkyl group).

) is reacted with the compound represented by the general formula (In the formula, R has the same meaning as above.

) or a salt thereof. R in the raw material 4 of the present invention means a lower alkyl group, such as methyl, ethyl, propyl, butyl, etc.

This raw material is prepared, for example, by reacting diethyl trans-epoxy succinate with an equimolar amount of alkali when it is cold. The raw material of the present invention is obtained, for example, by condensing NG-nitroagmatine and tert-butoxycarbonylleucine N-hydroxysuccinimide ester.

NG-nitro means that a nitro group is attached to the 2-position N of a guanidino group. In the first aspect of the present invention, a compound odor and a compound odor are mixed and reacted in the presence of triethylamine and diethyl phosphorocyanidate, usually in a solvent.

Examples of the solvent include dimethylformamide, dimethylacetamide, dimethylsulfoxide, hexamethylenephosphamide, and the like. Triethylamine forms the carboxylate anion of Compound 4, and utilizes the difference in basicity between the guanidino group and the amino group of the salt of Compound (3), such as dihydrochloride, to liberate only the amino group.
This is extremely convenient for selectively forming an acid amide bond with a compound (good) at an amino group, and the amount used is preferably 2.0 to 2.5 equivalents.

Although diethyl phosphorocyanidate forms an acid amide bond with an amino group and a carboxylic acid group, it can also be replaced with a known reagent for peptide synthesis, such as synchhexylcarbodiimide.
The reaction temperature is preferably in the range of -5°C to +35°C; high temperatures are not suitable because side reactions tend to occur. In the second aspect of the present invention, compound m[l] can be obtained by catalytic reduction of a compound (substitute) and then hydrolysis, or conversely, by catalytic reduction after hydrolysis.

Catalytic reduction can be performed in the presence of palladium on carbon, and hydrolysis can be performed with an inorganic acid, an organic acid, etc. used to hydrolyze ester bonds. It is appropriate to carry out these reactions at room temperature; raising the temperature to high temperatures tends to cause side reactions, which is not good. The present invention discloses a novel N-[N-(3-trans-carboxyoxirane-2-carbonyl)leucyl]agmatine {(M
The final product (1) of the present invention itself also has a thiol protease inhibitory effect.

Examples will now be described. Example 1 Production of ethyl hydrodiene trans-epoxy succinate (R = ethyl group) A solution of caustic potash 3.94V in ethanol (120 ml) was added to an ethanol (50 ml) solution of diethyl trans-epoxy succinate 13.1y at 5 to 10°C. When added under stirring, a solid precipitates out.

After further stirring at this temperature for 1 hour and then at room temperature for 1 hour, the monoethyl ester potassium salt of transepoxysuccinic acid precipitated was collected. (11.33f). After drying the mother liquor under reduced pressure, the residue was washed with ethyl acetate to obtain an additional 0.42 r of potassium salt as an insoluble matter. (Total 11.75f
7.85%). Elemental analysis samples were obtained by reprecipitation from water-ethanol. Coloring started at Mpl 55℃ and 283℃
The foam was decomposed. Elemental analysis Calculated value as C6H7O5K (%) C36.36, H3,56 experimental value (%)
C36. l3, H3.75NMR (D2O) δ: 1.
29 (3H.t, .J16.6Hz, .CH3), 3.
55 (2H, .ABq, .J2.0HZ1 oxirane ring proton), 4.25 (2H, q, .J6.6Hz, .
CH2). IR Dissolve the potassium salt thus obtained in 50 ml of water,
After acidifying with concentrated hydrochloric acid, the oily substance saturated with common salt and precipitated is extracted with ethyl acetate.

The ethyl acetate layers are combined, washed with saturated brine, dried over magnesium sulfate, ethyl acetate is distilled off, and the residue is fractionated under reduced pressure to obtain the desired product as a viscous oil. (7.8f7,7
0%). BpO. 3ll6~119℃ NMR (CDCl
3) δ: 1.32 (3H.t, J7.OHz,.CH3
), 3.70 (2H, .s1 oxirane ring proton),
4.26 (2H,.q,.J7.0Hz.CH2), 9
．． 85(1H,.s.C00H) Example 2N-(Te
Production of tert-butoxycarbonylleucine (tert-butoxycarbonylleucine)-N-hydroxysuccinimide ester 4.9
9y in 50 ml of dimethoxyethane solution under stirring with cooling.
A suspension is obtained by adding 9d of an aqueous solution of 3.90f of G-nitroagmatine hydrobromide and 1.69V of triethylamine.

This is stirred at room temperature for about 5.5 hours until the precipitate is dissolved.
The solution obtained is concentrated to about 15d and the residue is shaken with 40ml of 2.5% aqueous sodium carbonate solution and 40a of ethyl acetate. The aqueous layer is extracted with ethyl acetate, and the combined ethyl acetate layers are washed with saturated brine, 1 molar citric acid aqueous solution, and saturated brine, dried over magnesium sulfate, and ethyl acetate is distilled off.
The residue 6.48f is purified by silica gel column chromatography to give the product of this example (5.70r, 98%). This is recrystallized from a mixture of ethyl acetate and isopropyl ether. Mp9l~98℃ elemental analysis Cl6H32N6O,
Calculated values (%) as C49.47, H8.3O, N2l. 6
3 Experimental values (%) C49.6O, H8. l6, N2l.
4O Example 3 N-leucyl agmatine ([Production of O dihydrochloride (1) Method for first removing the nitro group and then the tert-butoxycarbonyl group N-(Tert-butoxycarbonyl leucyl) One repair One Nitro agmatine (substitute) 10% of 3.15f in a mixture of 40d methanol, 8ml acetic acid and 4d water.
In the presence of 0.4 f of palladium on carbon, the mixture is stirred and catalytically reduced under a weak hydrogen stream (about 4 hours).

▲ Separate the catalyst and wash with methanol, ▲ dry the washing liquid under reduced pressure. The residue was dissolved in 35d of water and extracted with ethyl acetate, and the aqueous layer was dried under reduced pressure to obtain the acetate salt of N-Tert-butoxycarbonylleucil-agmatine (2, 99f7). This was dissolved in 120 d of 99% formic acid and left to stand at room temperature for 3 hours, and then the formic acid was distilled off at 30°C or lower. The residue was dissolved in 40 d of water, extracted with ethyl acetate, and the aqueous layer was dried under reduced pressure to obtain N-leucyl agmatine diformate (2.58 f). Dissolve this in water and use the weakly acidic ion exchange resin Amberlite IR.
C5O (H+ form) is adsorbed onto a column packed with water, first eluted with water, and the eluate is dried to obtain 1.3 f of an oily substance.

Next, the column is eluted with 2N hydrochloric acid and the eluate is dried under reduced pressure to obtain N-leucyl agmatine ([I) dihydrochloride as a white solid (1.09f). Furthermore, the previously obtained oil was adsorbed onto the Amberlite IRC5O (H+ type) column again, and when eluted with water and 2N hydrochloric acid, N-
0.71f of leucyl agmatine (1) dihydrochloride is obtained. (Total 1.80f, 70%). This is methanol-
When recrystallized from an ether mixture, it becomes a white solid with a temperature of Mpl3O~137°C (decomposition). Elemental analysis C,,H2,Cl2
Calculated value for N and O (%) C4l. 77, H8.6l
, N22. l4 experimental value (%) C4l. 6O, H8.7
8, N22.342) Method for first removing Tert-butoxycarbonyl group and then nitro group N-(Tert
-butoxycarbonylleucyl)-NG-nitroagmatine (e) (5.70t) 99% formic acid (200Tf)
11) After leaving the solution at room temperature for 3 hours, formic acid is distilled off at a temperature below 30°C.

The residue was dissolved in 45 ml of water, extracted and washed with ethyl acetate, and the aqueous layer was dried under reduced pressure to obtain a residue of 5.351. Dissolve this in water and use the strongly acidic ion exchange resin Dowex.
x) Adsorb into a column packed with 50W (H+ type) and wash with water until the effluent becomes neutral. Next, the column was eluted with 2N hydrochloric acid, and the eluate was dried under reduced pressure to give N-leucyl-NG.
- Nitroagmatine hydrochloride is obtained as a viscous oil. This is dissolved in water 25WL1, and a strongly basic ion exchange resin DOwex 1 (0H-type) is added to neutralize it. After separating the resin from F, the solid (4.34 t) obtained by drying the liquid under reduced pressure was purified by silica gel column chromatography to obtain N-leucyl-NG-nitroagmatine as a solid (4.19 f198%). 1.50 f of N-leucyl-NG-nitroagmatine thus obtained is stirred in a weak hydrogen stream in the presence of 0.2 f of 10% palladium-carbon in a mixture of 25 a1 methanol, 6 d acetic acid, and 3 d water (4 hours).

▲ Separate the catalyst and wash with methanol ▲ Dry the washing liquid under reduced pressure.
The residue is dissolved in 20a of water and extracted with ethyl acetate, and the aqueous layer is dried under reduced pressure. 1.80f of the residue is dissolved in water and adsorbed on a column of weakly acidic ion exchange resin Amberlite IRC5O (H+ type), and first washed with water.

Next, the column was eluted with 2N hydrochloric acid and the eluate was dried under reduced pressure to obtain N-leucyl agmatine (11) dihydrochloride as a white solid (1.08V, 165%). Example 4 N-[N-(3-trans-ethoxycarbonyl-oxirane-2-carbonyl)monoleucyl]agmatine (1
．． Preparation of N-leucyl agmatine (I[) dihydrochloride 1.70f and ethylhydrodienetrans-epoxysuccinate (R=ethyl group)1.
0y was dissolved in dimethylformamide 11WL1, and diethyl phosphorocyanidate 1.0 was stirred at 0°C to -5°C.
A solution of 5y in dimethylformamide (2 ml) is added followed by a 2 ml solution of triethylamine 1.38V in dimethylformamide.

Claims (1)

[Claims] 1 N-leucyl agmatine having the general formula ▲There are mathematical formulas, chemical formulas, tables, etc.▼ (In the formula, R represents a lower alkyl group. ) is reacted with the general formula ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼ The general formula ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼ 1. A method for producing an N-leucyl agmatine compound or a salt thereof, which is characterized by obtaining a compound represented by: 2 Catalytic reduction and hydrolysis of the compound represented by the formula ▲There are mathematical formulas, chemical formulas, tables, etc.▼, and the resulting N-leucyl agmatine represented by the general formula ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼ (In the formula, R represents a lower alkyl group.) Compounds represented by the formula are reacted, and the general formula ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼ (R in the formula has the same meaning as above. A method for producing an N-leucyl agmatine compound or a salt thereof, characterized by obtaining a compound represented by: