KR100450607B1 - Process for preparing Galamin - Google Patents

Abstract

The present invention relates to a method for preparing a carmine derivative, and more particularly, aminoglyco by using sisomycin or gentamicin as a starting material with a smooth supply of raw materials, and easily introducing various derivatives into an amine group at the C-1 position. The present invention relates to a method for commercially and easily synthesizing a carmine derivative which is used as a key intermediate effective in the synthesis of the side antibiotics.

Description

Process for preparing galamine derivatives {Process for preparing Galamin}

The present invention relates to a method for preparing a carmine derivative, and more particularly, aminoglyco by using sisomycin or gentamicin as a starting material with a smooth supply of raw materials, and easily introducing various derivatives into an amine group at the C-1 position. The present invention relates to a method for commercially and easily synthesizing a carmine derivative which is used as a key intermediate effective in the synthesis of the side antibiotics.

Aminoglycoside antibiotics have been produced and commercially produced in a variety of aminoglycoside compounds since Waksman et al. Developed streptomycin antibiotics in 1944. Since then, neomycin in 1949, gentamicin in 1963, sisomicin in 1970, netilmicin in 1975, ispamicin in 1978, and arbekacin in 1990. 13-15 kinds of aminoglycoside antibiotics have been commercialized. The aminoglycoside antibiotics are highly toxic to liver and kidney. Aminoglycoside antibiotics inhibit the synthesis of the cell wall of bacteria and destroy the cell wall, which is much faster than other antibiotics.

Among the aminoglycoside antibiotics, some aminoglycoside antibiotics, including isepamycin, are relatively less toxic and complementary to or alone with antibiotics such as beta-lactam, glycopeptide and fluoroquinolone. It is known to be a very good medicine for infection. It also has an average 90% eradication rate for Gram-negative bacteria and Gram-positive bacteria.

In 1978, Schering protected the 6'-NH ₂ group with t-butyloxycarbonanyl azide, starting with gentamicin B obtained in small amount during fermentation of gentamicin in 1978, and then protecting the 1-NH ₂ group with N- (S- Aminoglycoside antibiotics, including isepamycin, were prepared by reacting with 4-benzyloxycarboylamino-L-hydroxybutyryloxy) succinimide and then removing them. J. Antibiotics , Nagabhushan, TL et al . , 1978 , 31 (7), 681-687. However, this method has a disadvantage in that the yield is very low.

Since then, Schering has developed a method that can selectively protect amines with similar reactivity in aminoglycosides, i.e., metal chelation, to significantly increase yields [ J. Am. Chem Soc ., Nagabhushan, TL et al . 1978 , 100 , 5253-5525. The method gentamicin a metal chelator that is B, Cu (Ⅱ), Ni (Ⅱ), Co (Ⅱ), Cd (Ⅱ) and reacted using a mixture thereof, other than the 1-NH ₂ group of NH ₂ group, Reaction of isoserine protected by chelation of this metal and substituted with 1-NH ₂ groups yields aminoglycoside compounds including substituted isepamycin and the protecting groups are removed to remove aminoglycosides including the desired isepamycin. A method for obtaining antibiotics has been developed (US Pat. No. 5,442,047). This method is a good method for producing isepamycin, but there are many problems in the supply of raw materials such as the yield of obtaining this compound in the fermentation process for obtaining gentamycin B is very low.

The inventors of the present invention sought a method for commercially synthesizing caramine derivatives as a key intermediate for the synthesis of various aminoglycoside antibiotics, including isepamycin, while supplying raw materials smoothly. In the meantime, N-benzyloxycarbonyl-S-isoserine including N-benzyloxycarbonyl-S-isoserine as a starting material is easy to supply raw material, such as sisomycin or gentamycin (mixture of gentamycin C ₁ , gentamycin C ₂ and gentamycin C _1a ). The present invention has been completed by establishing a new preparation method for synthesizing caramine derivatives by performing a process of removing a purpurosamine or a sosamine ring by a chemical method after binding the derivatives.

Accordingly, an object of the present invention is to provide a novel method for preparing carmine derivatives useful as a key intermediate for the synthesis of aminoglycoside antibiotics, including isepamycin.

The method for preparing a carmine derivative according to the present invention includes the following preparation process:

Agent for synthesizing a compound represented by the following Chemical Formula 1 by introducing an acetyl group (-Ac) or a benzeneoxycarbonanyl group (-Cbz) to an amine group of 3,2 ', 6'-NH ₂ of sisomycin or gentamicin 1 course,

A second process of synthesizing a compound represented by the following Chemical Formula 2 by coupling a amine group of 1-NH ₂ of the compound represented by Chemical Formula 1 with an anhydride of an acid represented by "YOH" or an acid derivative thereof,

A third step of synthesizing a compound represented by the following Chemical Formula 3 by introducing an acetyl group (-Ac) or a benzeneoxycarbonanyl group (-Cbz) to an amine group of 3 ″ -NH ₂ of the compound represented by Chemical Formula 2; And

Reacting the compound represented by the formula (3) under acidic conditions to remove the purpurosamine (purpurosamine) or the sosamine (sisosamine) ring to synthesize a carmine derivative represented by the following formula (4):

In Scheme 1, Represents a single bond or a double bond, R ₁ and R ₂ each represent a hydrogen atom or a methyl group, X represents an acetyl group (-Ac) or a benzeneoxycarbonanyl group (-Cbz), and Y represents 1 to 10 carbon atoms. An alkylcarbonyl group or an alkylcarbonyl group having 1 to 10 carbon atoms in which an amino or hydroxy group is substituted, and specifically, , , , , , , , , , , or Indicates.

Referring to the present invention in more detail as follows.

The present invention is characterized by using sisomycin or gentamicin (a mixture of gentamycin C ₁ , gentamycin C ₂ and gentamycin C _1a ) as starting materials. Sisomycin or gentamycin used in the present invention as a starting material have no problem in commercial application because of smooth supply of raw materials.

The process for preparing the carmine derivative according to the present invention will be described in more detail with reference to Scheme 1 as follows.

The first process selectively chelates the NH ₂ group of the starting material selected from sisomycin or gentamicin and adds an acetyl group (-Ac) or a benzeneoxycarbonyl group to the amine group of 3,2 ', 6'-NH ₂ . It is a process of synthesizing the compound represented by Chemical Formula 1 by introducing (-Cbz).

The amine group, hydroxy group, geminal or vicinal chelation of sisomycin or gentamicin used as starting material is optionally performed. In this case, Zn (OAc) ₂ · 2H ₂ O is used as a chelation reagent, and methanol, ethanol alcohol solvent, dimethylformamide (DMF), and dimethyl sulfoxide (DMSO) are used as the reaction solvent. The reaction for introducing an acetyl group (-Ac) or a benzeneoxycarbonanyl group (-Cbz) into the amine group (-NH ₂ ) at the C-3, C-2 'and C-6' positions is triethylamine or the like. In the presence of an inorganic base such as an organic amine base or sodium hydroxide. Acetic anhydride can be used as a reagent to introduce the acetyl group (-Ac), and N- (benzyloxycarbonyloxy) succinimide or benzyloxycarbonanyl chloride to introduce the benzeneoxycarbonyl group (-Cbz). May be used as a reagent. In particular, the reagent used for the benzyloxycarbonyl group introduction reaction has an advantage in that the reaction post-treatment process is relatively easy because it has excellent dissolution properties in organic solvents. The reaction of introducing the acetyl group (-Ac) or benzeneoxycarbonanyl group (-Cbz) is carried out at room temperature, the reaction is performed for about 6 to 24 hours 3,2 ', 6' represented by the formula (1) -Tri-N-substituted aminoglycosides are synthesized.

The second process is a process of synthesizing the compound represented by Chemical Formula 2 by combining a 1-NH ₂ group of the compound represented by Chemical Formula 1 with an anhydride of an acid represented by “YOH” or an acid derivative thereof. The acid compound represented by "YOH" may be substituted with an amino group or a hydroxy group as C ₁ to C ₁₀ alkylcarboxylic acid, and specifically 3-amino-2-hydroxypropionic acid and 4-amino-2-hydroxybutyric acid , Propionic acid, butyric acid, acetic acid, benzyloxycarboxylic acid, 5-amino-pentanoic acid, 5-hydroxypentanoic acid, 5-amino-heptanoic acid, 5-hydroxyheptanoic acid, formic acid, and the like. In the coupling reaction, alcohols such as methanol, DMF, DMSO, and the like are used as a reaction solvent. As the binder, dicyclohexyl carbodiimide (DCC), hydroxybenzotriazole (HOBT), 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide (EDCI) and the like are used.

The third process is a process of synthesizing the compound represented by Chemical Formula 3 by introducing an acetyl group (-Ac) or a benzeneoxycarbonanyl group (-Cbz) to an amine group having a position of C-3 ″. Acetyl group (-Ac ) Or a benzeneoxycarbonyl group (-Cbz) is introduced into a mixture of acetic anhydride or N- (benzyloxycarbonyloxy) succinimide or benzyloxycarbonyl chloride as described in the first step. Do the same in the same way.

In the fourth process, the compound represented by Chemical Formula 3 is removed by reacting the compound represented by Chemical Formula 3 under acidic conditions to remove a purpurosamine or sisosamin ring. It is a process. The reaction uses alcohols such as methanol, DMF, DMSO and the like as a reaction solvent, and sulfuric acid, hydrochloric acid, phosphoric acid, nitric acid and the like are used as an acid.

Meanwhile, the compounds represented by Chemical Formulas 1, 2, and 3 synthesized during the preparation process according to Scheme 1, and the caramine derivatives represented by Chemical Formula 4, which are intended for the present invention, are each novel compounds, and various aminoglycos Useful as an intermediate for synthesizing side antibiotics. Thus, the invention features these novel compounds as another feature.

The method for preparing a carmine derivative according to the present invention as described above will be described in more detail by the following examples, but the present invention is not limited by the following examples.

Example 1.

(1-1) Preparation of 3,2 ', 6'-tri-N-acetylcysomycin (Formula 1 , X = Ac)

2.57 g (5.8 mmol) of _sisomycin was dissolved in 29 mL of methanol, and 3.2 g (14.6 mmol) of Zn (OAc) _2˙ 2H ₂ O was added thereto, followed by stirring for 5 hours. A mixed solution of 4.1 mL (43.4 mmol) of triethylamine, 1.92 mL (20.4 mmol) of acetic anhydride and 30 mL of THF was added over 6 hours using a metering pump. The reaction mixture was stopped by adding concentrated aqueous ammonia solution (10 mL), and after distillation, the organic layer was extracted by distillation with chloroform, ethanol and concentrated aqueous ammonia solution, and distilled by column chromatography (CHCl ₃ / methanol / conc. NH ₄ OH = 3 /). 2/1).

Yield 80%; R _f 0.09 (CHCl ₃ / MeOH / conc. NH ₄ OH = 3/2 / 0.1); ¹ H NMR (300 MHz, D ₂ O) 5.32 (d, J = 1.99 Hz, 1H), 4.94 (d, J = 3.93 Hz, 1H), 3.96 (m, 2H), 3.92 (m, 4H), 3.45 (m, 2H), 3.16 (m, 2H), 2.73 (m, 1H), 2.67 (d, J = 10.76 Hz, 1H), 2.47 (s, 3H), 1.93 (s, 3H), 1.92 (m, 5H), 1.90 (s, 3H), 1.76 (s, 3H), 1.20 (s, 3H); ¹³ C NMR (75 MHz, D2O) 174.39, 174.08, 173.51, 165.21, 145.71, 100.86, 97.47, 96.32,86.83, 79.37, 75.40, 72.65, 69.55, 68.05, 63.65, 51.10, 48.22, 45.93, 41.59, 37.25, 34.28 , 31.75, 22.59, 22.36, 21.96; GC / LC / Mass (Mariner, ESI-TOF, mobile phase 50% acetone, 0.1% formic acid) m / z 136.0, 154.0, 307.0, 415.1, 574.1 (MH).

(1-2) N-benzyloxycarbonayl-L-isoserine

5 g (47.6 mmol) of L-isoserine and 7.5 g (54.3 mmol) of calcium carbonate were added to 30 mL of water, and 7.0 mL (50.0 mmol) of N-benzyloxycarbonyl chloride was slowly added thereto. After reacting at room temperature for 4 hours, the reaction was stopped with cold aqueous 2% hydrochloric acid solution, 100 mL of diethyl ether was added, and the obtained crystals were dissolved in ethyl acetate and recrystallized.

Yield 75%; R _f 0.63 (CHCl ₃ / MeOH / conc. NH ₄ OH = 3/2 / 0.3); ¹ H NMR (300 MHz, DMSO-d ₆ ) 7.60 to 7.22 (m, 5H), 5.18 (s, 2H), 4.05 (dd, J = 4.8, 7.05 Hz, 1H), 3.38 to 3.31 (m, 1H) , 3.30 to 3.14 (m, 1 H); ¹³ C NMR (75 MHz, DMSO-d ₆ ) 174.99, 157.07, 137.98, 129.20, 128.52, 70.21, 66.14, 45.20.

(1-3) Preparation of 1-[(3-N-benzyloxycarboyl-S-isoserine] -3,2 ', 6'-tri-N-acetyl sisomycin (Formula 2 )

0.1 g (0.17 mmol) of 3,2 ', 6'-tri-N-acetylcysomycin in 4 mL methanol, 41.7 mg (0.17 mmol) of 3-N-benzyloxycarboylisoserine, 29.1 mg (0.22 mmol) of HOBT , DCC63.5 mg (0.31 mmol) was added and stirred for 24 hours. After completion of the reaction, the solid formed was filtered and methanol was distilled off. Acidified with an aqueous 10% hydrochloric acid solution, chloroform was added to form a water layer, chloroform, ethanol, and concentrated aqueous ammonia solution were added thereto, and the organic layer was separated. The organic layer was distilled and a column chromatogram (CHCl ₃ / MeOH / conc. NH ₄ OH = 3/1 / 0.1) was performed.

Yield 70%; R _f 0.63 (CHCl ₃ / MeOH / conc. NH ₄ OH = 3/2 / 0.3); ¹ H NMR (300 MHz, D ₂ O) 7.29 (s, 5H), 5.37 (s, 1H), 5.01-4.96 (m, 3H), 4.13-3.19 (m, 2H), 2.49-2.36 (m, 4H ), 1.90-1. 8 (m, 14H), 1.26-1.25 (m, 1H), 1.07 (s, 3H); ¹³ C NMR (75 MHz, D ₂ O) 174.49, 174.17, 173.46, 158.78, 145.71, 136.72, 129.16, 128.78, 128.05, 99.08, 97.61, 95.38, 79.65, 78.79, 75.85, 72.52, 71.00, 69.05, 68.07, 6737 , 63.57, 49.95, 49.28, 47.93, 45.96, 44.01, 41.64, 36.95, 32.91, 22.63, 22.40, 22.29, 21.96; GC / LC / Mass (Mariner, ESI-TOF, mobile phase 95% acetone, 0.1%, formic acid, flow rate 10 μl / min) M / Z 116.1, 225.2, 398.2, 451.7, 795.1 (MH).

Example 2.

(2-1) Preparation of 3,2 ', 6'-tri-N-benzyloxycaraboylsisomicin (Formula 1 )

Dissolve 10 g (22.4 mmol) of _{sisomycin in} 110 mL of methanol, add 29.8 g (44.7 mmol) of Zn (OAc) _2˙ 2H ₂ O, stir for 6 hours, and then N- (benzyloxycarbonyloxy) succinimide. A solution of 19.5 g (78.3 mmol), 18.4 mL (0.13 mol) of triethylamine, and 80 mL of THF was added using a 6 hour metering pump. Stir for 24 hours, stop the reaction by adding a concentrated aqueous ammonia solution and methanol was distilled off. 45 mL of concentrated aqueous ammonia solution, 67 mL of ethanol and 100 mL of chloroform were added thereto, and the organic layer was separated and distilled to give a column chromatogram (CHCl ₃ / methanol / conc. NH ₄ OH = 5/1 / 0.1).

Yield 65%; R _f 0.33 (CHCl ₃ / MeOH / conc. NH ₄ OH = 5/1 / 0.1); ¹ H NMR (300 MHz, CDCl ₃ ) 7.56 to 7.30 (m, 15H), 5.38 (s, 1H), 5.11 to 4.98 (m, 7H), 4.53 (s, 1H), 3.83 to 3.65 (m, 8H) , 3.37 (s, 3H), 2.99 to 2.76 (m, 6H), 2.52 (s, 3H), 2.57 to 2.43 (m, 3H), 2.42 to 2.01 (m, 3H), 1.05 (s, 3H); ¹³ C NMR (75 MHz, CDCl ₃ ) 157.11, 156.51, 146.57, 136.94, 136.75, 128.92, 128.62, 128.46, 128.22, 101.98, 97.75, 96.70, 90.01, 76.21, 71.70, 69.61, 67.70, 69.61, 67.90, 67.00, 65.22, 50.63, 50.34, 47.89, 46.49, 43.03, 39.05, 37.30, 24.40, 23.09, 11.65; GC / LC / Mass (Mariner, ESI-TOF, mobile phase 95% acetone, 0.1% formic acid, flow rate 10 μl / min) m / z 416.7, 425.7, 670.3, 760.3, 850.3 (MH).

(2-2) Preparation of 1-[(3-N-benzyloxycarbonayl) -S-isoserine] -3,2 ', 6'-tri-N-benzyloxycarbonabosycisomycin (Formula 2 )

0.5 g (0.59 mmol) of 3,2 ', 6'-tri-N-benzyloxycaraboylsisomicin in 10 mL of methanol, 0.14 g (0.59 mmol) of 3- (N-benzyloxycarbonayl) isoserine, HOBT 0.1 g (0.76 mmol) and 0.22 g (1.1 mmol) of DCC were added thereto, and the resultant was stirred for 24 hours. After completion of the reaction, the solid formed was filtered and methanol was distilled off. Chloroform, ethanol and concentrated aqueous ammonia solution were added and the organic layer was separated. The organic layer was distilled and a column chromatogram (CHCl ₃ / MeOH / conc. NH ₄ OH = 5/1 / 0.1) was performed.

Yield 50%; R _f 0.38 (CHCl ₃ / MeOH / conc. NH ₄ OH = 5/1 / 0.1); ¹ H NMR (300 MHz, DMSO-d ₆ + CDCl ₃ ) 7.26 to 7.33 (m, 20H), 5.27 (s, 1H), 5.00 to 4.91 (m, 10H), 4.51 (s, 1H), 4.02 (s , 1H), 3.81 to 3.14 (m, 13H), 2.48 (s, 3H), 2.23 to 2.04 (m, 3H), 0.80 (s, 3H); GC / LC / Mass (Mariner, ESI-TOF, mobile phase 95% acetone, 0.1% formic acid, flow rate 10 μl / min) m / z 391.3, 527.2, 536.2, 1071.4 (MH).

(2-3) 1-[(3-N-benzyloxycaraboyl) -S-isoserine] -3,2 ', 6', 3 "-tetra-N-benzyloxycaraboylcysomycin (Formula 3 ) Manufacture

To 40 mL of THF and 0.12 mL (0.86 mmol) of triethylamine, 1-[(3-N-benzyloxycarbonanyl) -S-isoserine] -3,2 ', 6'-tri-N-benzyloxycarbonanyl 0.46 g (0.43 mmol) of sisomycin and 0.16 g (0.65 mmol) of N- (benzyloxycaraboyloxy) succinimide were added thereto and stirred for 24 hours. THF was distilled off, 50 mL of dichloromethane and 20 mL of 2% hydrochloric acid solution were added, the organic layer was separated, 40 mL of saturated sodium bicarbonate solution was added, and the organic layer was separated, filtered and dried. The compound was obtained.

Yield 70%; R _f 0.57 (ethyl acetate); ¹ H NMR (300 MHz, DMSO-d ₆ + CDCl ₃ ) 7.96 (s, 4H), 7.25 to 6.70 (m, 25H), 5.08 to 4.95 (m, 10H), 4.13 to 3.86 (m, 8H), 3.50 -3.00 (m, 3H), 2.87 (s, 3H), 2.07-2.04 (m, 2H), 1.22 (s, 3H); GC / LC / Mass (Mariner, ESI-TOF, mobile phase 95% acetone, 0.1% acetic acid, flow rate 10 μl / min) m / z 279.0, 283.0, 491.3, 634.2, 1205.4 (MH), 1222.5 (M + NH ₄ ) .

(2-4) Preparation of 1-[(3-N-benzyloxycaraboyl) -S-isoserine] -3,3'-di-N-benzyloxycarabornylamine (Formula 4 )

40.9 mg (62.3 mmol) of 1-[(3-N-benzyloxycaraboyl) -S-isoserine] -3,2 ', 6', 3 "-tetra-N-benzyloxycarboylcysomycin in 5 mL of ethanol PH was adjusted to 2 with sulfuric acid, stirred at room temperature for 5 hours, neutralized with saturated sodium bicarbonate solution and distilled methanol The residue was purified by column chromatography (CHCl ₃ / MeOH / conc. NH ₄ OH = 5). /1/0.1).

Yield 81%; R _f 0.17 (CHCl ₃ / MeOH / conc. NH ₄ OH = 5/1 / 0.1); ¹ H NMR (300 MHz, DMSO-d ₆ + CDCl ₃ ) 7.70 (s, 3H), 7.71-7.19 (m, 15H), 5.08-4.94 (m, 7H), 4.12-3.99 (m, 5H), 3.5 -3.2 (m, 3H), 2.87 (m, 3H), 1.42-1.39 (m, 1H), 0.90 (s, 3H); GC / LC / Mass (Mariner, ESI-TOF, mobile phase 95% acetone, 0.1% acetic acid, flow rate 10 μl / min) m / z 219.1, 307.1, 339.2, 409.2, 811.3 (MH), 828.3 (M + NH ₄ ) .

Example 3 Preparation of 1,3,3'-Tri-N-acetylgaramine from Sisomycin

(3-1) Preparation of 1,3,2 ', 6', 3 "-penta-N-acetylcysomycin

To 20 mL of acetone and methanol (1: 1, v / v) mixed solvent, 2.5 g (5.6 mmol) of sisomycin was added slowly to 20 mL of excess acetic anhydride (excess). After the reaction, the solvent and excess acetic anhydride were distilled off and the column chromatogram (CHCl ₃ / MeOH / conc. NH ₄ OH = 3/1 / 0.1) was obtained.

Yield 85%; R _f 0.17 (CHCl ₃ / MeOH / conc. NH ₄ OH = 3/1 / 0.1); ¹ H NMR (300 MHz, D ₂ O) 5.34 (3, 1H), 5.10 (d, J = 3.76 Hz, 1H), 4.02-3.42 (m, 10H), 3.19 (s, 2H), 2.95 (s, 3H), 2.04 (s, 3H), 1.92 (s, 3H), 1.87 (s, 3H), 1.81 (s, 3H), 1.78 (s, 3H), 0.88 (s, 3H); ¹³ C NMR (75 MHz, D ₂ O) 176.74, 176.45, 174.49, 174.14, 173.56, 145.60, 99.40, 99.15, 97.61, 96.20, 80.35, 78.53, 75.68, 75.52, 74.25, 73.96, 69.20, 64.68, 61.45, 56.09 , 49.61, 49.23, 47.93, 45.90, 41.61, 33.07, 32.39, 29.80, 22.58, 22.36, 22.24, 21.94, 21.00, 20.72.

(3-2) Preparation of 1,3,3'-tri-N-acetylgaramine (Formula 4 )

40.9 mg (62.3 mol) of 1,3,2 ', 6', 3 "-penta-N-acetylcysomycin was added to 5 mL of methanol, and the pH was adjusted to 2 with sulfuric acid. After stirring for 5 hours at room temperature, saturated bicarbonate Neutralize with sodium solution and distill the methanol The residue was subjected to column chromatography (CHCl ₃ / MeOH / conc. NH ₄ OH = 3/1 / 0.1).

Yield 65%; R _f 0.05 (CHCl ₃ / MeOH / conc. NH ₄ OH = 3/1 / 0.1); ¹ H NMR (300 MHz, D ₂ O) 5.15 (d, J = 3.83, 1H), 4.57 (d, J = 11.5 Hz, 1H), 4.53-3.21 (m, 8H), 2.87 (s, 1H), 2.11 (s, 3H), 1.91 (s, 3H), 1.83 (s, 3H), 0.95 (s, 3H).

Example 4 Preparation of 1,3,3'-Tri-N-benzyloxycarabornylamine from Sisomycin

(4-1) Preparation of 1,3,2 ', 6', 3 "-penta-N-benzyloxycarboylcysomycin

10 g (22.4 mmol) of sisomycin was dissolved in 20% aqueous sodium hydroxide solution (5.2 g of sodium hydroxide + 20 mL of water), and 16.6 mL (0.12 mol) of benzyl chloroformate was slowly added thereto and stirred for 24 hours. 300 mL of chloroform was added thereto, extracted, neutralized with ammonia chloride solution, filtered and dried and distilled to obtain a compound.

Yield 98%; R _f 0.1 (ethyl acetate / n-hexane = 2/1); ¹ H NMR (300 MHz, DMSO-d ₆ + CDCl ₃ ) 7.28 to 7.18 (m, 25H), 5.36 (s, 1H), 5.14 to 4.96 (m, 10H), 4.68 to 4.60 (m, 2H), 4.06 -3.52 (m, 13H), 3.02 (s, 3H), 2.03 (s, 3H); ¹³ C NMR (75 MHz, DMSO-d ₆ + CDCl ₃ ) 158.64, 157.25, 156.61, 137.33, 137.01, 128.89, 128.68, 128.45, 128.33, 127.86, 67.42, 66.87, 64.66, 59.11, 59.10, 48.00, 43.01, 30.59 , 22.14, 14.56.

(4-2) Preparation of 1,3,3'-tri-N-benzyloxycarabornylamine

To 100 mL of methanol was added 26 g (23.3 mmol) of 1,3,2 ', 6', 3 "-penta-N-benzyloxycarboylcysomycin, and the pH was adjusted to 2 with sulfuric acid. The mixture was stirred at room temperature for 5 hours. After neutralization with saturated sodium bicarbonate solution and methanol was distilled off. The residue was subjected to column chromatography (CHCl ₃ / MeOH / conc. NH ₄ OH = 5/1 / 0.1).

Yield 66%; R _f 0.24 (CHCl ₃ / MeOH / conc. NH ₄ OH = 5/1 / 0.1); ¹ H NMR (300 MHz, DMSO-d ₆ + CDCl ₃ ) 7.61 to 7.18 (m, 15H), 5.09 to 4.89 (m, 6H), 4.21 to 4.04 (m, 3H), 3.52 to 3.30 (m, 5H) , 3.28 to 2.99 (m, 2H), 2.38 (s, 3H), 1.27 to 1.10 (m, 2H); ¹³ C NMR (75 MHz, DMSO-d ₆ + CDCl ₃ ) 158.06, 157.78, 156.70, 138.12, 129.24, 129.16, 128.06, 127.69, 81.46, 75.18, 74.70, 73.95, 73.60, 69.85, 66.86, 66.04, 65.05, 59.42 , 52.11, 51.01, 31.52, 31.29, 23.22, 23.13.

Example 5 Preparation of 1-[(3-N-benzyloxycaraboyl-S-isoserine] -3,2 ', 6'-tri-N-acetylgentamycin (Formula 2 )

(5-1) Preparation of 3,2 ', 6'-tri-N-acetylgentamycin (Formula 1 )

10 g (21.6 mmol, average molecular weight 463.1) of gentamicin C2 was dissolved in 111 mL of methanol, and 11.8 g (54.0 mmol) of Zn (OAc) _2˙ 2H ₂ O was added thereto, followed by stirring for 5 hours. A mixed solution of 17.5 mL (0.13 mol) of triethylamine, 7.1 mL (75.8 mmol) of acetic anhydride, and 80 mL of THF was added over 6 hours using a metering pump. The reaction mixture was stopped by adding concentrated aqueous ammonia solution (10 mL), and after distillation, the organic layer was extracted by distillation with chloroform, ethanol and concentrated aqueous ammonia solution and distilled by column chromatography (CHCl ₃ / methanol: conc. NH ₄ OH = 3 /). 3 / 0.3).

Yield 80%; R _f 0.17-0.99 (CHCl ₃ / MeOH / conc. NH ₄ OH = 3/3 / 0.3).

(5-2) Preparation of 1-[(3-N-benzyloxycaraboyl-S-isoserine] -3,2 ', 6'-tri-N-acetylgentamycin (Formula 2 )

0.98 g (1.5 mmol) of 3,2 ', 6'-tri-acetylgentamycin, 0.37 g (1.5 mmol) of 3- (N-benzyloxycarbonanyl) isoserine, 0.27 g (2.0 mmol) of HOBT in 45 mL of methanol , DCC 0.57 g (2.8 mmol) was added and stirred for 24 hours. After completion of the reaction, the solid formed was filtered and methanol was distilled off. Acidified with 10% hydrochloric acid solution, chloroform was added to form a water layer, chloroform, ethanol and concentrated aqueous ammonia solution were added, and the organic layer was separated. The organic layer was distilled and a column chromatogram (CHCl ₃ / MeOH / conc. NH ₄ OH = 3/1 / 0.1) was performed.

Yield 70%

Example 6. Preparation of 1-[(3-N-benzyloxycaraboyl) -S-isoserine] -3,3'-di-N-benzyloxycarabonilgaramine

(6-1) Preparation of 3,2 ', 6'-tri-N-benzyloxycaraboylgentamycin (Formula 1 )

Dissolve 10 g (21.6 mmol) of gentamycin in 110 mL of methanol, add 11.8 g (54 mmol) of Zn (OAc) _2˙ 2H ₂ O, stir for 6 hours, and then add N- (benzyloxycarbonyloxy) succinimide. A solution of 18.8 g (75.8 mmol), 17.5 mL (0.13 mol) of triethylamine, and 80 mL of THF was added using a 6 hour metering pump. Stir for 24 hours, stop the reaction by adding a concentrated aqueous ammonia solution and methanol was distilled off. 45 mL of concentrated aqueous ammonia solution, 67 mL of ethanol and 100 mL of chloroform were added thereto, and the organic layer was separated and distilled to give a column chromatogram (CHCl ₃ / methanol: conc. NH ₄ OH = 5/1 / 0.1).

Yield 85%; R _f 0.37-0.39 (CHCl ₃ / MeOH / conc. NH ₄ OH = 5/1 / 0.1)

(6-2) Preparation of 1-[(3-N-benzyloxycarbonanyl) -S-isoserine] -3,2 ', 6'-tri-N-benzyloxycarbonabolygentamicin (Formula 2 )

2 g (2.3 mmol) of 3,2 ', 6'-tri-N-benzyloxycarbonylgentamycin in 40 mL of methanol, 0.55 g (2.3 mmol) of 3- (N-benzyloxycarboyl) isoserine, HOBT 0.4 g (3.0 mmol) and DCC 0.99 g (4.3 mmol) were added thereto, and the resultant was stirred for 24 hours. After completion of the reaction, the solid formed was filtered and methanol was distilled off. Chloroform, ethanol and concentrated aqueous ammonia solution were added and the organic layer was separated. The organic layer was distilled and a column chromatogram (CHCl ₃ / MeOH / conc. NH ₄ OH = 5/1 / 0.1) was performed.

Yield 75%

(6-3) 1-[(3-N-benzyloxycarbonyl) -S-isoserine] -3,2 ', 6', 3 "-tetra-N-benzyloxycaraboylgentamycin (Formula 3 ) Manufacture

To 50 mL of THF and 0.25 mL (1.8 mmol) of triethylamine, 1-[(3-N-benzyloxycarbonayl) -S-isoserine] -3,2 ', 6'-tri-N-benzyloxycarbonayl 1 g (0.92 mmol) of gentamicin and 0.35 g (1.4 mmol) of N- (benzyloxycaraboyloxy) succinimide were added thereto, followed by stirring for 24 hours. THF was distilled off, 50 mL of dichloromethane and 20 mL of 2% hydrochloric acid solution were added, the organic layer was separated, 40 mL of saturated sodium bicarbonate solution was added, and the organic layer was separated, filtered and dried. Got.

Yield 80%

(6-4) Preparation of 1-[(3-N-benzyloxycaraboyl) -S-isoserine] -3,3'-di-N-benzyl-oxycarabornylamine (Formula 4 )

0.5 g (0.41 mol) of 1-[(3-N-benzyloxycarbonyl) -S-isoserine] -3,2 ', 6', 3 "-tetra-N-benzyloxycaraboylgentamycin in 60 mL methanol PH was adjusted to 2 with sulfuric acid, stirred at room temperature for 5 hours, neutralized with saturated sodium bicarbonate solution and distilled methanol The residue was purified by column chromatography (CHCl ₃ / MeOH / conc. NH ₄ OH = 5). /1/0.1).

Yield 87%, the same as the physicochemical data of Example 2.

Example 7 Preparation of 1,3,3'-Tri-N-acetylgaramine (Formula 4 ) from Gentamicin

(7-1) Preparation of 1,3,2 ', 6', 6 ', 3 "-penta-N-acetylgentamycin

1.57 g (39.3 mmol) of sodium hydroxide powder was added to 50 mL of acetone, 10 g of gentamicin sulfate was added, the reaction temperature was lowered to 0 ° C., 20 mL of excess acetic acid (excess) was slowly added, and the reaction temperature was raised to room temperature for 24 hours. Stir it. Acetic anhydride and acetone were added in an excess amount, and 50 mL of concentrated ammonia water, 50 mL of ethanol and 200 mL of chloroform were added thereto, and the organic layer was separated. The organic layer was dried, filtered and distilled to obtain a compound.

Yield 61%; R _f 0.41 to 0.43 (CHCl ₃ / MeOH / conc. NH ₄ OH = 3/1 / 0.1)

(7-2) Preparation of 1,3,3'-tri-N-acetylgaramine (Formula 4 )

0.1 g of 1,3,2 ', 6', 6 ', 3 "-penta-N-acetylgentamycin was added to 10 mL of methanol, and the pH was adjusted to 2 with sulfuric acid. After stirring for 5 hours at room temperature, saturated sodium bicarbonate was added. The solution was neutralized and methanol was distilled off The residue was subjected to column chromatography (CHCl ₃ / MeOH / conc. NH ₄ OH = 5/1 / 0.1).

Yield 70%; R _f 0.05 (CHCl ₃ / MeOH / conc. NH ₄ OH = 3/1 / 0.1); Same as the physicochemical data of Example 3.

Example 8 Preparation of 1,3,3'-Tri-N-benzyloxycarabornylamine from Gentamicin

(8-1) Preparation of 1,3,2 ', 6', 3 "-penta-N-benzyloxycaraboylgentamycin

14.4 g (0.36 mmol) of sodium hydroxide powder was added to a mixed solvent of acetonitrile (50 mL) and water (50 mL), 10 g of gentamicin sulfate was added thereto, and the reaction temperature was reduced to 0 ° C. and 15.3 mL (0.11 mol) of benzyl chloroformate was added. ) Was added slowly and the reaction temperature was raised to room temperature and stirred for 24 hours. Acetonitrile was distilled off, extracted with chloroform, filtered and dried to obtain a compound.

Yield 96%; R _f 0.11 to 0.19 (ethyl acetate / n -hexane = 2/1)

(8-2) Preparation of 1,3,3'-tri-N-benzyloxycarabornylamine

1 g of 1,3,2 ', 6', 3 "-penta-N-benzyloxycarcarbonylgentamycin was added to 20 mL of methanol, and the pH was adjusted to 2 with sulfuric acid. After stirring for 5 hours at room temperature, saturated sodium bicarbonate was added. The solution was neutralized and methanol was distilled off The residue was subjected to column chromatography (CHCl ₃ / MeOH / conc. NH ₄ OH = 5/1 / 0.1).

Yield 66%; R _f 0.24 (CHCl ₃ / MeOH / conc. NH ₄ OH = 5/1 / 0.1); Same as the physicochemical data of Example 4.

Example 9. Preparation of 1-N-isobutyryl-3,2 ', 6'-tri-N-acetylcysomycin (Formula 4 )

To 3 mL of DMF, add 50 mg (87.3 mol) of 3,2 ', 6'-tri-N-acetylcysomycin and 36.5 μL (0.26 mmol) of triethylamine, and slowly add 15.9 μL (96.0 μmol) of isobutyric acid. gave. After completion of the reaction, DMF and triethylamine were distilled off and the column chromatogram (CHCl ₃ / MeOH / conc. NH ₄ OH = 3/1 / 0.1) was obtained.

Yield 85%; R _f 0.43 (CHCl ₃ / MeOH / conc. NH ₄ OH = 3/1 / 0.1); ¹ H NMR (300 MHz, D ₂ O) 5.38 (s, 1H), 4.98 (d, J = 3.6 Hz, 1H), 4.00-3.50 (m, 9H), 3.20 (d, J = 12.39 Hz, 1H) , 2.56 (d, J = 10.53 Hz, 1H), 2.42 (s, 3H), 1.99 (s, 3H), 1.91 (s, 3H), 1.80 (s, 3H), 1.50-1.35 (m, 1H), 1.10 (s, 3 H), 0.97 (d, J = 6.84 Hz, 6 H); HR-MS (FAB ⁺ ) m / z 160.1, 211.1, 485.2, 644.3 (M−H).

Example 10. Preparation of 1-[(3-N-benzyloxycarboyl-S-homoserine] -3,2 ', 6'-tri-N-benzyloxycarbonabosysisomicin (Formula 4 )

(10-1) N-benzyloxycarbonayl-L-homoserine

1 g (8.4 mmol) of L-homoserine and 3.5 g (25.2 mmol) of calcium carbonate were added to 10 mL of water, and 1.3 mL (9.2 mmol) of N-benzyloxycarbonyl chloride was slowly added thereto. After reacting at room temperature for 6 hours, the reaction was stopped with cold aqueous 2% hydrochloric acid solution and 40 mL of diethyl ether was added to precipitate a solid.

Yield 98%; R _f 0.31 (CHCl ₃ / MeOH / conc. NH ₄ OH = 3/2 / 0.1); ¹ H NMR (300 MHz, CDCl ₃ + DMSO-d ₆ ) 7.47 to 7.29 (m, 5H), 5.04 (s, 2H), 4.14 to 4.07 (m, 1H), 3.50 to 3.41 (m, 2H), 1.88 -1.85 (m, 1H), 1.84-1.67 (m, 1H); ¹³ C NMR (75 MHz, CDCl ₃ + DMSO-d ₆ ) 175.07, 157.00, 137.80, 129.09, 128.52, 66.23, 58.12, 51.79, 34.64.

(10-2) Preparation of 1-[(3-N-benzyloxycaraboyl-S-homoserine] -3,2 ', 6'-tri-N-benzyl-oxycaraboylcysomycin

0.3 g (0.35 mmol) of 3,2 ', 6'-tri-N-benzyloxycaraboylsisomicin in 10 mL methanol, 89.5 mg (0.35 mmol) of 3-N-benzyloxycarbonayl-L-homoserine, HOBT 61.2 mg (0.45 mmol) and DCC 0.14 mg (0.66 mmol) were added and stirred for 24 hours. After completion of the reaction, the solid formed was filtered and methanol was distilled off. Acidified with an aqueous 10% hydrochloric acid solution, chloroform was added to form a water layer, chloroform, ethanol, and concentrated aqueous ammonia solution were added thereto, and the organic layer was separated. The organic layer was distilled and a column chromatogram (CHCl ₃ / MeOH / conc. NH ₄ OH = 9/1 / 0.1) was performed.

Yield%; R _f 0.16 (CHCl ₃ / MeOH / conc. NH ₄ OH = 9/1 / 0.1); ¹ H NMR (300 MHz, CDCl ₃ + DMSO-d ₆ ) 7.55 to 7.33 (m, 20H), 4.25 to 4.20 (m, 2H), 3.57 to 3.37 (m, 7H), 4.98 to 4.77 (m, 8H) , 4.25 to 4.20 (m, 2H), 3.57 to 3.37 (m, 7H), 2.98 to 2.90 (m, 7H), 2.51 to 2.50 (m, 2H), 2.42 to 2.27 (m, 1H), 1.95 to 1.17 ( m, 5H), 1.14 (s, 3H).

As of 1998, aminoglycoside compounds with worldwide sales of around 200 billion are effective against many types of bacteria and are less resistant than other antibiotics, and thus are used as complementary drugs with other antibiotics. The future development of antibiotics is expected because the market for antibiotics is rapidly gaining accurate information about bacteria in the clinic, thanks to the development of genomics and protenomics and the remarkable developments in the biochip field. The market for aminoglycoside compounds is expected to grow steadily because it is expected to move toward the development of drugs that are resistant to both bacteria and one or two types of bacteria, rather than the development of drugs that inhibit all bacteria.

The key intermediates and caramine derivatives produced by the present invention can be prepared at low cost and used for the synthesis of aminoglycoside compounds, including isepamycin.

The 1994 Cload et Amino the side-based compound is a lot of research since the combination with high affinity with HIV RNA Rev Responsive element showed a development potential of Sharon cloud HIV inhibitors it proceeds glycosides for neomycin B (IC ₅₀ = 0.1 to 1 μM) shows potential as an HIV inhibitor, but is seeking new compounds because of the toxicity of neomycin B itself. In this study, compounds having 1,3-hydroxyamine or 1,3-diamine groups as functional groups in aminoglycoside compounds were known to bind with high affinity to HIV RNA Rev Responsive element. The development of new HIV inhibitors is also expected when synthesizing new compounds.

Claims (11)

A process for synthesizing a compound represented by the following Chemical Formula 1 by introducing an acetyl group (-Ac) or a benzeneoxycarboyl group (-Cbz) to an amine group of 3,2 ', 6'-NH ₂ of sisomycin or gentamicin , A process of synthesizing a compound represented by the following Chemical Formula 1 by coupling a amine group of 1-NH ₂ of the compound represented by Chemical Formula 1 with an anhydride of an acid represented by "YOH" or an acid derivative thereof; Introducing an acetyl group (-Ac) or a benzeneoxycarbonanyl group (-Cbz) to an amine group of 3 "-NH ₂ of the compound represented by Formula 2 to synthesize a compound represented by the following Formula 3, and Reacting the compound represented by the formula (3) under acidic conditions to remove the purpurosamine (purpurosamine) or the sosamine (sisosamine) ring to synthesize a carmine derivative represented by the following formula (4) Preparation of Derivatives: In the above scheme, Represents a single bond or a double bond, R ₁ and R ₂ each represent a hydrogen atom or a methyl group, X represents an acetyl group (-Ac) or a benzeneoxycarbonanyl group (-Cbz), and Y represents 1 to 10 carbon atoms. An alkylcarbonyl group having 1 to 10 carbon atoms in which an alkylcarbonyl group or an amino or hydroxy group is substituted. The method of claim 1, wherein the first step is to react with Zn (OAc) ₂ · 2H ₂ O to selectively chelate NH ₂ group of sisomycin or gentamicin, and then acetyl group (-Ac) or benzeneoxycara Method for producing a carbonyl group (-Cbz) characterized by introducing. The method of claim 1, wherein the acetyl group (-Ac) or benzeneoxycarbonanyl group (-Cbz) introduction reaction in the first or third process is performed in the presence of an organic base or an inorganic base. . 4. The acetic acid anhydride is used for introduction of the acetyl group (-Ac), and N- (benzyloxycarboyloxy) succinimide or benzyloxycarbonayl is introduced for the introduction of the benzeneoxycarbonyl group (-Cbz). Process for producing a chloride characterized in that the use. The method of claim 1, wherein the acid represented by "YOH" in the second step is 3-amino-2-hydroxypropionic acid, 4-amino-2-hydroxybutyric acid, propionic acid, butyric acid, acetic acid, benzyloxycarboxylic acid, And anhydrides or derivatives thereof of the acid selected from 5-amino-pentanoic acid, 5-hydroxypentanoic acid, 5-amino-heptanoic acid, 5-hydroxyheptanoic acid and formic acid. The method of claim 1, wherein the coupling reaction in the second step is dicyclohexylcaraboimide (DCC), hydroxybenzotriazole (HOBT) and 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide (EDCI) A process for producing a binder, characterized in that using. The method according to claim 1, wherein the acid in the fourth process is selected from sulfuric acid, hydrochloric acid, phosphoric acid and nitric acid. delete delete delete delete