KR100615893B1 - Process for the preparation of rr,ss-4,8-diaza-3,6,6,9-tetramethylundecane-2,10-dione bisoxime derivatives - Google Patents

Abstract

The present invention is a method for synthesizing a propylene amine oxime derivative, a compound used to make a complex of technetium-99m, a useful radionuclide for imaging and diagnosing organs in vivo. A method for synthesizing the d, l isomers of a compound in a clean state without contamination of unwanted meso isomers as needed.

Propyleneamine Oxime Derivative, Technetium-99m

Description

(Al, S) -4,8-diaza-3,6,6,9-tetramethyl undecane-2,10-dione bis oxime derivative manufacturing method {PROCESS FOR THE PREPARATION OF (RR, SS) -4 , 8-DIAZA-3,6,6,9-TETRAMETHYLUNDECANE-2,10-DIONE BISOXIME DERIVATIVES}

(RR, SS) -4,8-diaza-3,6,6,9-tetramethylundecane of the formula (15) named d, l-HMPAO (d, l-Hexamethyl propyleneamine oxime) derivative A method for producing a 2,10-dione bisoxime derivative purely without contamination of other meso isomers.

Wherein R is a lower alkyl group with carbon atoms can be arranged with one to six straight or branched chain, - (CH ₂₎ n- or a ring shape that can be represented by _{- (CH 2) aX- (CH} 2) b- It means a ring form that can be represented by. In this case, X means N, O, S, and the like, and n, a, and b are independent variables, which mean 1 to 10, respectively. R ₁ means a lower alkyl group which may be arranged in straight or branched chain having 1 to 6 carbon atoms, and R ₂ is a lower alkyl group or lower egg which may be arranged in a straight or branched chain having 1 to 10 carbon atoms Or a lower alkyl group or a lower alkane group. R ₃ means H or a lower alkyl group or benzyl group which may be arranged in a straight or branched chain having 1 to 6 carbon atoms.

(RR, SS) -4,8-diaza-3,6,6,9-tetramethylundecane-2,10 of Formula 15 designated as d, l-HMPAO (d, l-Hexamethyl propyleneamine oxime) derivative -Dion bisoxime derivatives and technetium-99m complexes are distributed through the blood-brain-barrier and distributed to the brain in proportion to the blood volume of the brain. Make it possible. Among the stereoisomers of HMPAO, the d, l isomer has better biodistribution rate in the brain than the meso isomer, and the l isomer is better among the d, l isomers.

In the present invention, two compounds having optical activity are used as starting materials, and the reaction selectivity is excellent, and the reaction conditions are mild, so that the manufacturing process is convenient, the yield of each step is relatively high, and the purity of the meso isomer is high (RR, SS) -4,8-diaza-3,6,6,9-tetramethylundecane-2,10-dione bisoxime derivative.

The conventional method for preparing d, l-HMRAO is based on 2,3-butanedione monooxime and 2,2-dimethyl-1,3-propanediamine as starting materials, and first, HMPAO as a mixture of d, l isomers and meso isomers. After this, a number of fractional crystallizations are performed therefrom to obtain d, l-HMPAO. This method is described in EP 194,843 and the basic synthesis method can be summarized in Scheme 1.

First, 2 equivalents of 2,3-butanedione monooxime is reacted with 2,2-dimethyl-1,3-propanediamine to obtain an imine form of a compound, followed by sodium borohydride to form HMPAO. Since the hydride attack of sodium borohydride can occur freely up and down, the obtained HMPAO is obtained in the form of a mixture of three isomers. The less meso isomer of the imaging effect can be separated from d, l-HMPAO by a continuous fractional crystallization method and can be separated by HPLC method.

Both of these purification methods have big disadvantages. First, the continuous fractional crystallization method requires at least 7 recrystallization steps to obtain pure d, l-HMPA0, resulting in long working hours and very low yields. In addition, the separation method by HPLC has similar separation time between meso-HMPAO and d, l-HMPAO, resulting in poor separation efficiency and difficulty in separating large amounts of samples even with Prep-HPLC. As a result, the manufacturing method of d, l-HMPAO shown in EP 194,843 has the disadvantage of separation method that is difficult to prepare a large amount of samples.

An object of the present invention is (RR, SS) -4,8-diaza-3,6,6,9-tetramethylundecane-2,10, generally designated as d, l-HMPAO derivative and represented by the formula (15). It is to provide a method for producing dione bisoxime derivatives in high yield without the production of meso-HMPAO, and the production process is convenient, eliminating the multiple fractional crystallization method used in the existing method, and high purity and mass production It is to provide a method of manufacturing by the process that becomes possible.

In order to achieve the above object, in the present invention (RR, SS) -4,8-diaza-3,6,6,9-tetramethylundecane-2 represented by the following d, l-HMPAO derivative Provided is a method for producing a, 10-dione bisoxime derivative in high purity and high yield by an industrially convenient manufacturing process.

The (RR, SS) -4,8-diaza-3,6,6,9-tetramethylundecane-2,10-dione bisoxime derivative represented by Chemical Formula 15 of the present invention is represented by the following Scheme 2. It is manufactured and can be used as an imaging diagnostic reagent for diagnosing organs in vivo by measuring cerebral blood flow by forming a complex mixture with Technetium-99m. (J. Nucl. Med. 28, 191, 1987)

Wherein R is a lower alkyl group with carbon atoms can be arranged with one to six straight or branched chain, - (CH ₂₎ n- or a ring shape that can be represented by _{- (CH 2) aX- (CH} 2) b- It means a ring form that can be represented by. In this case, X means N, O, S, and the like, and n, a, and b are independent variables, which mean 1 to 10, respectively. R ₁ means a lower alkyl group which may be arranged in a straight or branched chain having 1 to 6 carbon atoms. P is an acyl halide derivative capable of combining with a secondary amine to form an amide group, R ₂ is a lower alkyl group, lower alken group, lower alka group which can be arranged in a straight or branched chain having 1 to 10 carbon atoms. Popularity, etc. R ₃ means H or lower alkyl or benzyl groups which may be arranged in straight or branched chains having 1 to 6 carbon atoms.

A diester compound of Formula 8a and 8b is prepared by reacting a malonic acid derivative such as Formula 5 with a D-alanine methyl ester hydrochloride derivative of Formula 6 and an L-alanine methyl ester hydrochloride derivative of Formula 7, respectively. The two compounds of Formula 8a and 8b are mixed in a 1: 1 weight ratio, and the mixture is reacted with a reducing agent such as lithium aluminum hydride to form a dialcohol compound which is a mixture of Formulas 9a and 9b. Protecting the two amine groups of this compound with a suitable protecting group to prepare a dialcohol compound which is a mixture of the formulas 10a and 10b in which the two amine groups of the mixture of formulas 9a and 9b are protected, and then reacts the two alcohol groups of the compound with a suitable oxidant. To prepare a dialdehyde compound which is a mixture of Formulas 11a and 11b. A suitable Grignard reagent is reacted with the prepared dialdehyde compound to prepare a dialcohol compound, which is a mixture of Chemical Formulas 12a and 12b, and then reacted with a suitable oxidant to prepare a diketo compound, which is a mixture of Chemical Formulas 13a and 13b. . This diketo compound is reacted with a hydroxylamine derivative in the presence of an acid to give a high yield of the dioxime compound, which is a mixture of formulas 14a and 14b. Finally, the protecting group may be separated by a suitable deprotecting reagent to prepare a desired compound which is a mixture of Formula 15a and Formula 15b.

Structural Characteristics and Activity Relationships of d, l-HMPAO

The combination of Technetium-99m pertechnetate and d, l-HMPAO in aqueous solution creates the Technetium-99m lipophilic macrocyclic complex, which is used as a diagnostic agent for radiation. This mixture has zero net-charge centers and OHO hydrogen bonding, is easily adapted to radionuclide ligands and is stable enough for parenteral administration, making it very useful for imaging the body. Can be used.

The expected structure of the mixture is represented by the following Chemical Formula 16,

The expected structure of a compound useful as a radiation diagnostic reagent is a lipophilic macrocyclic complex of propyleneamine oxime ligand of formula 17 and technetium-99m as follows.

The compound of formula 17 shown in the above figure has two sub-carbons and there are three stereoisomers, and the in-vivo properties of the complex of these stereoisomers and technetium-99m There is a big difference between them.

In Scheme 3, Formula 17 shows meso-diastereomers, which are optically inactive, and d- and l-diastereomers, which are optically active, d and l. Isomers occur because carbons 3 and 9 are sub carbons. Meso-diastereomers have very different properties from d, l-diastereomers (racemic mixtures of d- and l-mirrors).

For example, they have different melting points and different separation times on HPLC. In addition, d- and l- mirror images of the general formula (17) have different physical images and biological properties from mirror images, racemic mixtures of mirror images, and mesodiastereomers. The difference in in vivo properties of the mixture of the two diastereomers and technetium-99m is very different. As a result, the difference in cerebral absorption between the two isomers is surprising. In the case of Formula 17, the d, l isomer is 2.3 times better than the meso isomer, and similarly, the l isomer is 1.6 times better than the d isomer.

Structural isomers (cis isomers and trans isomers), on the other hand, may exist in three different isomeric forms because there may be limited rotation about the C = N bond of the two oxime groups. Each isomer has different physical properties (eg m.p, b.p) and can be separated by chromatographic techniques (TLC, HPLC). Their interconversion is generally fluid and can be catalyzed by mineral acids, Lewis acids, bases, metal ions.

A typical preparation example according to the preparation method of the present invention is shown in Scheme 4. Looking at Scheme 4, the formula 15a 'and the formula 15b' through 8-step from the 2,2-dimethyl malonic acid represented by the formula (5 ') And (RR, SS) -4,8-diaza-3,6,6,9-tetramethylundecane-2,10-dione bisoxime as a mixture.

As mentioned earlier, the first step of the reaction used the conditions of George R. Pettit (J. Org. Chem. 50, 2323, 1985) as the amide formation reaction. The intermediate obtained by the reaction of 2,2-dimethyl malonic acid and isobutylchloroformate was reacted with D-alanine methyl ester hydrochloride and L-alanine methyl ester hydrochloride having optical activity, respectively, to formulas (8a ') and (8b'). Compounds of can be obtained in high yield, respectively. The second step of the reaction used the conditions of Jean-Claude Fiaud (J. 0rg. Chem. 52, 5320, 1987) as the reduction reaction. Dialcohol, which is a mixture of Formula 9a 'and Formula 9b', in which two amide groups and two ester groups are reduced in a reduction reaction of a mixture of two ester compounds of Formulas 8a 'and 8b' obtained in a 1: 1 weight ratio. Compounds can be obtained. The third step of protecting the two amine groups consists of protecting groups of Boc or protecting groups of CBZ according to a known method, in which a dialcohol compound is obtained which is a mixture of formulas 10a 'and 10b' with good yield. The next 4 to 5 steps are performed as follows according to a known method. According to Daniel Swern (J. 0rg. Chem. 43, 2480, 1978), the oxidation reaction of two primary alcohols of the dialcohol compounds of the formulas (10a ') and (10b') is carried out in good yield without epimerization. A dialdehyde compound is obtained, which is a mixture of 'and Formula 11b', which is reacted with Grignard reagent of methylmagnesium bromide according to known methods (Tetrahedron Lett. 29, 6905, 1988). And a compound of a secondary alcohol which is a mixture of the formula 12b '. Again, the compounds of Formula 12a 'and Formula 12b' undergo a second oxidation reaction according to the conditions of Daniel Swern mentioned above to obtain a diketo compound which is a mixture of Formula 13a 'and Formula 13b' with good yield. The oxime molding reaction is reacted with hydroxylamine in the presence of an acid according to a known patent (W0 97/38000) to obtain an oxime compound which is a mixture of the formulas (14a ') and (14b'). To be obtained. In the final eighth reaction step, a protecting group is removed by catalyzed hydrogen reaction in the presence of palladium / carbon to obtain a mixture of Formula 15a 'and Formula 15b', which is the desired d, l-HMPA0. Most of the reaction of the present invention has the advantage of obtaining d, l-HMPAO with high purity because the reaction yield of each step is relatively high, the manufacturing process is convenient, and no meso-HMPAO is produced.

Hereinafter, the preparation method of the present invention will be described in detail with reference to Scheme 4.

Ⅰ. First Step

(2R, 8R) -3,7-diaza-5,5-dimethyl-2,8-dimethoxycarbonylnonan-4,6-dione and (2S, 8S) -3,7-diaza-5, Preparation of 5-dimethyl-2,8-dimethoxycarbonylnonan-4,6-dione

In the present invention, the amidation reaction of carboxylic acid and amine such as 2,2-dimethyl malonic acid and alanine methyl ester hydrochloride is performed by protic solvents such as methylene chloride, chloroform, carbon tetrachloride, dimethylformamide, dimethyl sulfoxide and the like. It can be carried out in the same aprotic solvent and the activation reaction of the carboxylic acid is ethyl in the presence of a base such as triethylamine, N, N-diisopropylamine, 4-dimethylaminopyridine, pyridine, N-methylmorpholine, morpholine, etc. It can be produced by reacting with chloroformate, isobutylchloroformate, t-butylchloroformate, trimethylacetyl chloride and the like. The reaction temperature is preferably -20 ° C to 50 ° C, but more preferably -5 ° C to 10 ° C. In the reaction between the intermediate and the amine, the solvent used in the intermediate production process is used as it is, and at least one equivalent base is used to neutralize the hydrochloride salt of the amine.

Amide production reaction temperature is suitable from 0 ℃ to 50 ℃ and the reaction time is preferably from 3 hours to 24 hours. The reaction can also be purified cleanly by recrystallization or silica gel chromatography after a simple work up. By this amide formation reaction, (2R, 8R) -3,7-diaza-5,5-dimethyl-2,8-dimethoxycarbonylnonane-4,6-dione is obtained from D-alanine methyl ester hydrochloride. (2S, 8S) -3,7-diaza-5,5-dimethyl-2,8-dimethoxycarbonylnoan-4,6-dione is obtained from L-alanine methyl ester hydrochloride. .

II. 2nd step

Preparation of (RR, SS) -3,7-diaza-2,5,5,8-tetramethyl-1,9-dihydroxynonane

(2R, 8R) -3,7-diaza-5,5-dimethyl-2,8-dimethoxycarbonylnonan-4,6-dione and (2S, 8S)-represented by formulas (8a ') and (8b')- A 1: 1 mixture of 3,7-diaza-5,5-dimethyl-2,8-dimethoxycarbonylnonane-4,6-dione is reacted with a strong reducing agent lithium aluminum hydride in a suitable solvent to react the two amides. Group and two ester groups are reduced to (RR, SS) -3,7-diaza-2,5,5,8-tetramethyl-1,9-dihydroxynonane. Diethyl ether, tetrahydrofuran, etc. can be used as a solvent here, It is especially preferable to use the tetrahydrofuran solution. The reaction temperature is a suitable reflux temperature, the reaction time is 10 to 30 hours is good, but 20 to 25 hours is particularly suitable. This reaction can also be used as such in the third stage reaction described below without special purification, such as recrystallization or silica gel chromatography after a simple work up.

III. 3rd step

Preparation of (RR, SS) -N, N-dibenzyloxycarbonyl-3,7-diaza-2,5,5,8-tetramethyl-1,9-dihydroxynonane

(RR, SS) -3,7-diaza-2,5,5,8-tetramethyl-1,9-dihydroxynonane represented by a mixture of formulas 9a 'and 9b' has two amine groups protected Is converted to (RR, SS) -N, N-dibenzyloxycarbonyl-3,7-diaza-2,5,5,8-detramethyl-1,9-dihydroxynonane. As the protecting group, methoxycarbonyl chloride, 2,2,2-trichloroethoxycarbonyl chloride, di-t-butyl dicarbonate, allyloxycarbonyl chloride, benzyl chloroformate, 4-nitrobenzyloxycarbonyl chloride Etc. may be used and it is particularly preferable to use di-t-butyldicarbonate or benzylchloroformate. Examples of the base include sodium bicarbonate, sodium carbonate, sodium hydroxide, potassium hydroxide, pyridine, and the like, and sodium carbonate is particularly preferable. As the reaction solvent, tetrahydrofuran / H ₂ O, acetonitrile / H ₂ O, dioxane / H ₂ O, dimethylformamide / H ₂ O, dimethyl sulfoxide / H ₂ O, etc. are used, and tetrahydrofuran / Preference is given to using H ₂ O or dioxane / H ₂ O. The reaction temperature is preferably 0 ° C. to 50 ° C., but more preferably from room temperature to 30 ° C., and the reaction time is preferably from 3 hours to 30 hours, but more preferably from 5 hours to 10 hours. In addition, this reaction can be used for the next reaction through a purification process such as silica gel chromatography after a simple work up, or can be used as it is without purification.

Ⅳ. 4th step

Preparation of (RR, SS) -N, N-dibenzyloxycarbonyl-3,7-diaza-5,5-dimethyl-2,8-diformylnonane

(RR, SS) -N, N-dibenzyloxycarbonyl-3,7-diaza-2,5,5,8-tetramethyl-1,9-di represented by a mixture of formulas 10a 'and 10b' Two hydroxy groups of hydroxynonane are oxidized to have (RR, SS) -N, N-dibenzyloxycarbonyl-3,7-diaza-5,5-dimethyl-2,8-di having two aldehyde groups Converted to formylnonane. In the present invention, a pyridium chloro chromate reaction or a Swern reaction, which is a typical oxidation reaction, was performed.

Reaction 1

Methylene chloride, chloroform, etc. may be used as the reaction solvent, and chromium trioxide 2 pyridine / celite, pyridium chloro chromate / sodium acetate / molic sieve, and pyridium dichromate may be used as the reaction reagent. Preference is given to using pyridium chlorochromate / sodium acetate / molic acid.

The reaction temperature can be 10 ℃ to 40 ℃ but it is better to react at room temperature, the reaction time is 30 minutes to 3 hours, but it is more preferable to terminate between 30 minutes to 1 hour. After completion of the reaction, it is important to remove excess pyridium chloro chromate by adding diethyl ether or diisopropyl ether. This reaction can be used in the next reaction with or without special purification, such as silica gel chromatography after filtration through celite.

Reaction 2

Methylene chloride, chloroform, etc. may be used as the reaction solvent, and dimethyl sulfoxide / oxalyl chloride / triethylamine, dimethyl sulfoxide / oxalyl chloride / N, N-diisopropylethylamine, and dimethyl sulfoxide may be used as reaction reagents. / Dimethylsulfonic anhydride / triethylamine, dimethyl sulfoxide / thionyl chloride / triethylamine and the like can be used, in particular to use dimethyl sulfoxide / oxalyl chloride / N, N- diisopropylethylamine desirable. The reaction temperature can be from -78 ° C to room temperature, but it is more preferable to terminate the reaction between -78 ° C and -10 ° C, and the reaction time is preferably completed within 3 to 5 hours. This reaction is followed by purification, such as silica gel chromatography after a simple work up, and then used in the next reaction.

Ⅴ. 5th step

Preparation of (RR, SS) -N, N-dibenzyloxycarbonyl-4,8-diaza-3,6,6,9-tetramethyl-2,10-dihydroxyundecane

(RR, SS) -N, N-dibenzyloxycarbonyl-3,7-diaza-5,5-dimethyl-2,8-diformylnonane represented by the mixture of formulas 11a 'and 11b' is 2 Nucleophilic attack of Grignard reagent with aldehyde groups resulted in (RR, SS) -N, N-dibenzyloxycarbonyl-4,8-diaza- having two secondary alcohol groups Is converted to 3,6,6,9-tetramethyl-2,10-dihydroxyundecane. Diethyl ether, methylene chloride, tetrahydrofuran, etc. may be used as the reaction solvent, and tetrahydrofuran is preferably used. The reaction reagent may be prepared by using Grignard reagent using methyl bromide and magnesium pieces, and preferably, commercially available lignard reagent of a suitable concentration may be used. The reaction temperature can be from -78 ° C to room temperature, but it is preferable to react from -60 ° C to 0 ° C, and the reaction time is preferably performed within 30 minutes to 2 hours. In addition, this reaction shows three spots on TLC, and after simple work up, it is important to separate all three spots through purification process such as silica gel chromatography.

Ⅵ. 6th step

Preparation of (RR, SS) -N, N-dibenzyloxycarbonyl-4,8-diaza-3,6,6,9-tetramethylundecane-2,10-dione

(RR, SS) -N, N-dibenzyloxycarbonyl-4,8-diaza-3,6,6,9-tetramethyl-2,10-di represented by a mixture of formulas 12a 'and 12b' Hydroxyundecane is (RR, SS) -N, N-dibenzyloxycarbonyl-4,8-diaza-3,6,6,9- by Swern oxidation among the two methods of the fourth step described above. Converted to tetramethylundecane-2,10-dione. As the reaction reagent, it is preferable to use dimethyl sulfoxide / oxalyl chloride / N, N-diisopropylethylamine among the reagents listed in step 4, and the reaction temperature is preferably performed at room temperature at -78 ° C. This reaction is followed by purification, such as silica gel chromatography after a simple work up, and then used in the next reaction.

Iii. 7th Step

Preparation of (RR, SS) -N, N-dibenzyloxycarbonyl-4,8-diaza-3,6,6,9-tetramethylundecane-2,10-dione bisoxime

(RR, SS) -N, N-dibenzyloxycarbonyl-4,8-diaza-3,6,6,9-tetramethylundecane-2,10 represented by a mixture of formulas 13a 'and 13b' -Dione is a (RR, SS) -N, N-dibenzyloxycarbonyl-4,8-diaza-3,6,6,9-tetra in which two ketone groups react with hydroxylamine to have two oxime groups Is converted to methylundecane-2,10-dione bisoxime. Methanol, ethanol, isopropanol, butanol and the like may be used as the reaction solvent, but methanol is preferably used. As the reaction reagent, an aqueous solution of hydroxylamine, hydroxylamine hydrochloride, etc. may be used. As the acid catalyst, acetic acid, formic acid, propionic acid, and the like may be used. The reaction temperature may be from room temperature to 70 ° C., but is preferably from 50 ° C. to 60 ° C., and the reaction time is preferably from 2 hours to 5 hours. In this reaction, the oxime mainly has a trans form, and a small amount of cis form can be removed by a purification process such as silica gel chromatography after a simple work up.

Iii. 8th Step

Preparation of (RR, SS) -4,8-diaza-3,6,6,9-tetramethylundecane-2,10-dione bisoxime

(RR, SS) -N, N-dibenzyloxycarbonyl-4,8-diaza-3,6,6,9-tetramethylundecane-2,10 represented by a mixture of Formulas 14a 'and 14b' -Dione bisoxime is converted to (RR, SS) -4,8-diaza-3,6,6,9-tetramethylundecane-2,10-dione bisoxime by two hydrogen amine protecting groups separated by hydrogenation. do. Methanol, ethanol, isopropanol, ethyl acetate, tetrahydrofuran, etc. may be used as the reaction solvent, and ethanol is preferable. As a reaction reagent, palladium / carbon is used as a catalyst for general hydrogenation reaction under hydrogen pressure of appropriate atmospheric pressure, and palladium / carbon is used as catalyst for phase transfer hydrogenation reaction and 1,3- is used as a hydrogen source. Cyclohexadiene, 1,4-cyclohexadiene, ammonium formate and the like. In the present invention, a hydrogenation reaction was performed using palladium / carbon and hydrogen or 1,4-cyclohexadiene with palladium / carbon. The reaction temperature is suitably 20 ° C. to 40 ° C., and the reaction time is preferably between 2 hours and 5 hours. In addition, the reaction was concentrated after a simple filtration through celite, and the concentrated solution was recrystallized using a solvent such as ethyl acetate / hexane, methanol / ethyl acetate / hexane, methanol / diethyl ether / ethyl acetate, and (RR, SS) -4,8-diaza-3,6,6,9-tetramethylundecane-2,10-dione bisoxime (d, l-HMPAO) can be obtained.

In the present invention, the molecular structure was confirmed by infrared spectroscopy, UV-visible spectroscopy, nuclear magnetic resonance spectrum, spectrophotometry, mass spectrometry, elemental analysis of representative compounds and comparison of actual values. The purity of the compound was also confirmed by HPLC analysis.

Hereinafter, the present invention will be described in more detail with reference to Examples. However, the following examples are merely to illustrate the content of the present invention is not limited to the scope of the present invention.

Example

<Example 1>

(2R, 8R) -3,7-diaza-5,5-dimethyl-2,8-dimethoxycarbonylnonan-4,6-dione and (2S, 8S) -3,7-diaza-5, Preparation of 5-dimethyl-2,8-dimethoxycarbonylnonan-4,6-dione

2,2-dimethyl malonic acid (5 g, 37.8 mmol) was dissolved in 400 ml of methylene chloride and cooled to 0 to 5 DEG C under a hobbing bath. 4-methylmorpholine (9.15ml, 83.2mmol) and isobutylchloroformate (10.8m1, 83.2mmol) were slowly added dropwise to the solution, followed by reaction at 0 ° C to 5 ° C for 2 hours. 4-methylmorpholine (9.15ml, 83.2mmol) was further added to this solution, and then D-alanine methyl ester hydrochloride (1.16g, 83.2mmol) was added to 50 ml of methylene chloride. After removing the ice bath, the reaction solution was heated to room temperature and stirred for 24 hours. 300 ml of saturated aqueous citric acid solution was added to the reaction solution, followed by extraction with methylene chloride. The organic layer is washed with saturated aqueous sodium bicarbonate solution and saturated brine and then dehydrated with anhydrous magnesium sulfate (MgSO ₄ ). After filtration, the filtrate was concentrated and the concentrate was subjected to silica gel column chromatography (solvent; ethyl acetate: n-hexane = 3: 1) to obtain the target compound (2R, 8R) -3,7-diaza-5,5-dimethyl. 9.38 g (82%) of -2,8-dimethoxycarbonylnonane-4,6-dione are obtained.

Similarly, the above reaction process was carried out using L-alanine methyl ester and hydrochloride, and the target compound (2S, 8S) -3,7-diaza-5,5-dimethyl-2,8-dimethoxycarbonylnonane Obtain 9.42 g (82%) of -4,6-dione.

¹ H-NMR (20 MHz, CDCl ₃ ): δ 7.04 (bd, 2H), 4.49 (m, 2H), 3.72 (s, 6H),

1.44 (s, 6H), 1.39 (d, 6H)

IR (neat): 3350.6, 2992.5, 1748.7, 1673.3, 1522.5, 1221.0 cm ^-1

<Example 2>

Preparation of (RR, SS) -3,7-diaza-2,5,5,8-tetramethyl-1,9-dihydroxynonane

Lithium aluminum hydride (10 g, 263.5 mmol) was added to 150 ml of anhydrous tetrahydrofuran and cooled to 0 ° C to 5 ° C under an ice bath. (2R, 8R) -3,7-diaza-5,5-dimethyl-2,8-dimethoxycarbonylnonan-4,6-dione (4.25 g, 14.05 mmol) and (2S, 8S) -3,7-diaza-5,5-dimethyl-2,8-dimethoxycarbonylnonan-4,6-dione (4.25 g, 14.05 mmol) was dissolved in 100 ml of anhydrous tetrahydrofuran, and this solution Add carefully dropping funnels using a dropping funnel. The ice bath is removed and refluxed for 20 hours. The reaction mixture was cooled to _{0 ℃ ~5 ℃ H 2 O 10ml} , 15% sodium hydroxide aqueous solution and then the side deeper 20ml, note the H ₂ O 10ml was added slowly and then the mixture was stirred for 20 minutes. The resulting aluminum mixture was filtered through celite and washed with 50 ml of tetrahydrofuran. The filtrate was concentrated and dried in vacuo to give 6.1 g (99%) of the title compound.

¹ H-NMR (200 MHz, CDCl ₃ ): δ 3.53 (dd, 2H), 3.24 (dd, 2H), 2.72 (m, 2H),

2.63-2.26 (m, 4H), 0.99 (d, 6H), 0.88 (s, 6H)

IR (neat): 3319.7, 2964.7, 1469.8, 1373.4, 1153.5 cm ^-1

<Example 3>

Preparation of (RR, SS) -N, N-di-t-butoxycarbonyl-3,7-diaza-2,5,5,8-tetramethyl-1,9-dihydroxynonane

(RR, SS) -3,7-diaza-2,5,5,8-tetramethyl-1,9-dihydroxynonane (198 mg, 0.91 mmol) obtained in Example 2 was added with 4 ml of tetrahydrofuran and H. was dissolved in a mixed solvent of ₂ 0 4ml, then sodium carbonate (577mg, 5.4mmol) and di -t- butyl dicarbonate (0.63ml, 2.72mmol) was stirred for 15 hours at room temperature the reaction solution was added. After completion of the reaction, 50 ml of ethyl acetate is added and the mixture is washed with saturated citric acid solution, saturated aqueous sodium bicarbonate solution and saturated brine, and the organic layer is dehydrated with anhydrous magnesium sulfate. After filtration, the filtrate was concentrated and the concentrate was purified by silica gel column chromatography (solvent; ethyl acetate: n-hexane = 3: 1) to obtain 302 mg (79.5%) of the title compound.

¹ H-NMR (200 MHz, CDCl ₃ ): δ 3.81 (m, 4H), 3.22-3.12 (m, 6H), 1.43 (s, 18H),

1.28 (d, 6H), 0.90 (s, 6H)

IR (neat): 3451.7, 2979.3, 1290.4, 1216.2 cm ^-1

<Example 4>

Preparation of (RR, SS) -N, N-di-t-butoxycarbonyl-3,7-diaza-5,5-dimethyl-2,8-diformylnonane

(RR, SS) -N, N-di-t-butoxykirbonyl-3,7-diaza-2,5,5,8-tetramethyl-1,9-dihydroxynonane obtained in Example 3 (278mg, 0.66mmol) was dissolved in 25ml of methylene chloride, followed by addition of sodium acetate (1.63g, 19.9mmol), finely divided molecular sieve 3Å (2g), pyridiumchlorochromate (2.86g, 13.3mmol) at room temperature The reaction solution is stirred vigorously for 50 minutes. After completion of the reaction, 100 ml of diethyl ether was added thereto, stirred for 20 minutes, and the resulting solid was filtered through Celite. The filtrate was concentrated and the residue was dissolved in 50 ml of ethyl acetate, washed with saturated aqueous sodium bicarbonate solution, and the organic layer was dehydrated with anhydrous magnesium sulfate and filtered. The residue obtained by concentrating the filtrate was subjected to silica gel column chromatography (solvent; ethyl acetate: n-hexane = 1: 1.5) to obtain 193 mg (70.1%) of the title compound.

¹ H-NMR (200 MHz, CDCl ₃ ): δ 9.66 (s, 2H), 3.70-3.30 (m, 4H), 3.85 (m, 2H),

1.43 (d, 6H), 1.38 (s, 18H), 1.01 (s, 6H)

IR (neat): 3379.0, 2965.4, 1702.4, 1278.4 cm ^-1

Example 5

Preparation of (RR, SS) -N, N-di-t-butoxycarbonyl-4,8-diaza-3,6,6,9-tetramethyl-2,10-dihydroxyundecane

(RR, SS) -N, N-di-t-butoxycarbonyl-3,7-diaza-5,5-dimethyl-2,8-diformylnonane (160 mg, 0.39 mmol) obtained in Example 4 ) Is dissolved in 10 ml of tetrahydrofuran and cooled to -78 ° C using a dry ice / acetone bath. Take 3M-methylmagnesium bromide (1.3ml, 3.9mmol) into the syringe and drop it slowly into the gastric mixture. After stirring the reaction solution at -78 ° C for 1 hour, saturated aqueous citric acid solution is added and the reaction temperature is slowly raised to room temperature. 50 ml of ethyl acetate is added to separate the organic layer, and the organic layer is washed with saturated aqueous sodium bicarbonate solution and saturated brine, and then dehydrated with anhydrous magnesium sulfate. The filtrate was concentrated after filtration, and the residue was purified by silica gel column chromatography (solvent; ethyl acetate: n-hexane = 0.8: 1) to obtain 115 mg (66.7%) of the title compound.

¹ H-NMR (20 MHz, CDCl ₃ ): δ 4.14 (m, 2H), 3.34 (m, 2H), 2.98-2.69 (m, 4H),

1.60-0.90 (m, 36H)

IR (neat): 3425.9, 2984.2, 1690.7, 1289.0 cm ^-1

<Example 6>

Preparation of (RR, SS) -N, N-di-t-butoxycarbonyl-4,8-diaza-3,6,6,9-tetramethylundecane-2,10-dione

12 ml of methylene chloride was added to the reaction vessel and cooled to -78 ° C using a dry ice / acetone bath. Oxalyl chloride (0.16 ml, 1.83 mmol) and dimethyl sulfoxide (0.24 ml, 3.38 mmol) were slowly added dropwise to this solution.

(RR, SS) -N, N-di-t-butoxycarbonyl-4,8-diaza-3,6,6,9-tetramethyl- obtained in Example 5 after stirring at −78 ° C. for 20 minutes. A solution of 2,10-dihydroxyundecane (99 mg, 0.22 mmol) dissolved in 5 ml of methylene chloride is slowly added dropwise and stirred at this temperature for 20 minutes. N, N-diisopropylethylamine (0.8 ml, 4.59 mmol) was added to this reaction solution, and the reaction temperature was raised from -78 ° C to room temperature. Saturated citric acid solution is added to this solution, followed by extraction with methylene chloride. The organic layer was washed with saturated aqueous sodium bicarbonate solution and saturated brine, dehydrated with anhydrous magnesium sulfate, filtered and the filtrate was concentrated. The concentrate was subjected to silica gel column chromatography (solvent; ethyl acetate: n-hexane = 1: 1.5) to obtain 87.6 g (90%) of the target compound.

¹ H-NMR (20 MHz, CDCl ₃ ): δ 3.60-3.05 (m, 6H), 2.19 (s, 6H), 1.59 (s, 6H),

1.38 (s, 18H), 1.06 (s, 6H)

IR (neat): 2974.5, 1710.9, 1420.4, 1278.0 cm ^-1

<Example 7>

Preparation of (RR, SS) -N, N-di-t-butoxycarbonyl-4,8-diaza-3,6,6,9-tetramethylundecane-2,10-dione bisoxime

(RR, SS) -N, N-di-t-butoxycarbonyl-4,8-diaza-3,6,6,9-tetramethylundecane-2,10-dione obtained in Example 6 ( 97 mg, 0.22 mmol) was dissolved in 7 ml of methanol, 50% aqueous hydroxylamine solution (0.44 ml, 6.6 mmol) and acetic acid (3.4 ml, 2.25 mmol) were added sequentially, followed by stirring at 50 ° C. for 3 hours. After completion of the reaction, the reaction solution was cooled to room temperature and 30 ml of ethyl acetate was added thereto. After separating the organic layer, the organic layer is washed with saturated citric acid solution, saturated aqueous sodium bicarbonate solution and saturated salt in that order. The organic layer is dehydrated with anhydrous magnesium sulfate, filtered and concentrated. The concentrated solution was subjected to silica gel column chromatography (solvent; ethyl acetate: n-hexane = 1: 1.5) to obtain 75 mg (72.5%) of the title compound.

¹ H-NMR (200 MHz, CDCl 3): δ 3.90 (m, 2H), 3.16 (m, 4H), 1.87 (s, 6H),

1.45 (d, 6H), 1.39 (s, 18H), 0.97 (s, 6H)

IR (neat): 3378.4, 2935.0, 1700.4, 1514.9, 1270.4 cm ^-1

<Example 8>

Preparation of (RR, SS) -4,8-diaza-3,6,6,9-tetramethylundecane-2,10-dione bis oxime

(RR, SS) -N, N-di-t-butoxycarbonyl-4,8-diaza-3,6,6,9-tetramethylundecane-2,10-dione bis obtained in Example 7 Trifluuroacetic acid (0.41 ml, 5.3 mmol) was added to a solution of oxime (250 mg, 0.53 mmol) in 10 ml of methylene chloride, followed by stirring at room temperature for 5 hours. When the starting material on the TLC disappeared, the pH of the reaction solution was adjusted to 8-9 using a saturated aqueous sodium bicarbonate solution and extracted with methylene chloride. The extract is dehydrated with anhydrous magnesium sulfate, filtered and concentrated. 1 ml of ethyl acetate was added to the concentrate, followed by stirring for 10 minutes. Then, 2 ml of hexane was added thereto, stirred for 20 minutes, filtered, and the obtained solid was dried in vacuo to yield 65 mg (45%) of the title compound.

¹ H-NMR (200 MHz, CDCl 3): δ 3.25 (q, 2H), 2.25 (q, 4H), 1.72 (s, 6H),

1.72 (s, 6H), 0.82 (s, 6H)

IR (KBr): 3237.5, 1452.5, 1361.8, 978.0, 837.2 cm ^-1

Example 9

Preparation of (RR, SS) -N, N-dibenzyloxycarbonyl-3,7-diaza-2,5,5,8-tetramethyl-1,9-dihydroxynonane

(RR, SS) -3,7-diaza-2,5,5,8-tetramethyl-1,9-dihydroxynonane (6.2 g, 2.84 mmol) obtained in Example 2 was added to 70 ml of tetrahydrofuran. After dissolving, a solution of sodium carbonate (9 g, 8.49 mmol) was added to 40 ml of H ₂ O. After cooling the solution to 0 ° C.-5 ° C. under an ice bath, benzylchloroformate (16.1 ml, 11.3 mmol) was slowly added dropwise using a dropping funnel. After completion of the dropwise addition, the ice bath was removed and the reaction solution was stirred at room temperature for 15 hours. After completion of the reaction, 200 ml of ethyl acetate was added, washed with saturated citric acid solution, saturated aqueous sodium bicarbonate solution and saturated brine, and then the organic layer was dehydrated with anhydrous magnesium sulfate. After filtration, the filtrate was concentrated and the concentrate was purified by silica gel column chromatography (solvent; ethyl acetate: n-hexane = 5: 1) to obtain 11.0 g (80.2%) of the title compound.

¹ H-NMR (200 MHz, CDCl ₃ ): δ 7.26 (s, 1OH), 5.04 (s, 4H), 3.77 (m, 4H),

3.40-2.85 (m, 6H), 1.23 (s, 6H), 8.86 (s, 6H)

IR (neat): 3447.0, 2978.2, 1678.1, 1290.3, 1215.2, 1026.1 cm ^-1

<Example 10>

Preparation of (RR, SS) -N, N-dibenzyloxycarbonyl-3,7-diaza-5,5-dimethyl-2,8-diformylnonane

(Example 10-1)

(RR, SS) -N, N-dibenzyloxycarbonyl-3,7-diaza-2,5,5,8-tetramethyl-1,9-dihydroxynonane (9.28 g) obtained in Example 9 , 19.1 mmol) was dissolved in 500 ml of methylene chloride, followed by addition of sodium acetate (47 g, 572.9 mmol), finely divided molecular sieve 3 g (30 g), pyridium chloro chromate (41 g, 190.2 mmol), and the reaction solution at room temperature for 50 minutes. Stir vigorously. After completion of the reaction, 700 ml of diethyl ether was added thereto, stirred for 20 minutes, and the resulting solid was filtered through Celite. The filtrate was concentrated, and the residue was dissolved in 200 ml of ethyl acetate, washed with saturated aqueous sodium bicarbonate solution, and the organic layer was dehydrated with anhydrous magnesium sulfate and filtered. The residue obtained by concentrating the filtrate was subjected to silica gel column chromatography (solvent; ethyl acetate: n-hexane = 1: 1.5) to obtain 6.91 g (75.1%) of the title compound.

¹ H-NMR (200 MHz, CDCl ₃ ): δ 9.60 (m, 2H), 7.29 (s, 10H), 5.08 (s, 4H),

3.70-3.20 (m, 4H), 3.70-3.20 (m, 4H), 3.10-2.70 (m, 2H),

1.30 (m, 6H), 0.90 (m, 6H)

IR (neat): 3378.9, 2964.3, 1701.6, 1428.6, 1277.7 cm ^-1

(Example 10-2)

100 ml of methylene chloride was added to the reaction vessel, which was then cooled to -78 ° C using a dry ice / acetone bath. Oxalyl chloride (2.5 ml, 28.0 mmol) and dimethyl sulfoxide (3.86 ml, 54.4 mmol) were added to the solution. Slowly add dropwise. (RR, SS) -N, N-dibenzyloxycarbonyl-3,7-diaza-2,5,5,8-tetramethyl-1,9 obtained in Example 9 after stirring at −78 ° C. for 20 minutes. -A solution of dihydroxynonane (1.8 g, 3.7 mmol) dissolved in 30 ml of methylene chloride is slowly added dropwise and stirred at this temperature for 30 minutes. After adding N, N-di isopropylethylamine (12 ml, 68.9 mmol), the reaction temperature was raised from -78 ° C to -5 ° C. At this temperature, saturated aqueous citric acid solution is added and the temperature is raised to room temperature. Methylene chloride was added to the mixture, followed by extraction. The organic layer was washed with saturated aqueous sodium bicarbonate solution and saturated brine, and then dried over anhydrous magnesium sulfate, filtered and concentrated. The concentrated solution was purified by silica gel column chromatography (solvent; ethyl acetate: n-hexane = 1.5: 1) to obtain 1.62 g (91%) of the title compound. The assay value of the compound was the same as that of the compound obtained in Example 10-1. It was.

<Example 11>

Preparation of (RR, SS) -N, N-dibenzyloxycarbonyl-4,8-diaza-3,6,6,9-tetramethyl-2,10-dihydroxyundecane

(RR, SS) -N, N-dibenzyloxycarbonyl-3,7-diaza-5,5-dimethyl-2,8-diformylnonane (5.6 g, 11.6 mmol) obtained in Example 10 was prepared. Dissolve in 170 ml of tetrahydrofuran and cool to -78 ° C using a dry ice / acetone bath. 3M-Methylmagnesium bromide (39ml, 116mmol) is taken by syringe and slowly added dropwise to the gastric mixture. The reaction solution was stirred at -78 ° C for 1 hour, and then saturated aqueous citric acid solution was added to gradually raise the reaction temperature to room temperature. 300 ml of ethyl acetate is added to separate the organic layer, and the organic layer is washed with saturated aqueous sodium bicarbonate solution and saturated brine, and then dehydrated with anhydrous magnesium sulfate. The filtrate was concentrated after filtration, and the residue was purified by silica gel column chromatography (solvent; ethyl acetate: n-hexane = 1: 1.2-&gt; 2.5: 1) to obtain 3.88 g (65%) of the title compound.

¹ H-NMR (200 MHz, CDC1 ₃ ): δ 7.28 (s, 10H), 5.03 (s, 4H), 4.04 (m, 2H),

3.32 (m, 2H), 2.78 (m, 4H), 1.16 (d, 6H), 1.06 (s, 6H),

0.80 (d, 6H)

IR (neat): 3426.0, 2983.1, 1691.9, 1286.9, 1060.8 cm ^-1

<Example 12>

Preparation of (RR, SS) -N, N-dibenzyloxycarbonyl-4,8-diaz-3,6,6,9-tetramethylundecane-2,10-dione

150 ml of methylene chloride was added to the reaction vessel, followed by cooling to -78 ° C using a dry ice / acetone bath. Oxalyl chloride (4.2 ml, 48.97 mmol) and dimethyl sulfoxide (6.3 ml, 88.69 mmol) were slowly added dropwise to this solution. (RR, SS) -N, N-dibenzyloxycarbonyl-4,8-diaza-3,6,6,9-tetramethyl-2,10 obtained in Example 11 after stirring at −78 ° C. for 20 minutes. -A solution of dihydroxyundecane (3.09 g, 6.0 mmol) dissolved in 70 ml of methylene chloride is added dropwise and stirred at this temperature for 20 minutes. N, N-diisopropylethylamine (20 ml, 117.6 mmol) was added to the reaction solution, and the reaction temperature was raised from -78 ° C to room temperature. Saturated citric acid solution is added to the solution, followed by extraction with methylene chloride. The organic layer is washed with saturated aqueous sodium bicarbonate solution and saturated brine, dehydrated with anhydrous magnesium sulfate, filtered and the filtrate is concentrated. Silica gel column chromatography (solvent; ethyl acetate: n-hexane = 1: 1.5-&gt; 1.5: 1) was used to obtain 2.9 g (95%) of the target compound.

¹ H-NMR (200 MHz, CDC1 ₃ ): δ 7.27 (s, 1OH), 5.03 (s, 4H), 3.40-2.80 (m, 6H),

2.18-1.80 (m, 6H), 1.6O-1.20 (m, 6H), 1.10-0.90 (m, 6H)

IR (neat): 2973.7, 1711.0 1418.9, 1277.5 cm ^-1

Example 13

Preparation of (RR, SS) -N, N-dibenzyloxycarbonyl-4,8-diaza-3,6,6,9-tetramethylundecane-2,10-dione bis oxime

(RR, SS) -N, N-dibenzyloxycarbonyl-4,8-diaza-3,6,6,9-tetramethylundecane-2,10-dione (3 g, 5.87) obtained in Example 12 mmol) was dissolved in 100 ml of methanol, 50% hydroxylamine aqueous solution (11.5 ml 187.7 mmol) and acetic acid (3.4 ml, 59.4 mmol) were added sequentially, followed by stirring at 50 ° C. for 3 hours. After completion of the reaction, the reaction solution was cooled to room temperature and 200 ml of ethyl acetate was added thereto. After separating the organic layer, the organic layer is washed with saturated citric acid solution, saturated aqueous sodium bicarbonate solution and saturated salt in that order. The organic layer is dehydrated with anhydrous magnesium sulfate, filtered and concentrated. This concentrate was carefully silica gel column chromatographed (solvent; ethyl acetate: n-hexane = 1: 2) to obtain 2.3 g (72.4%) of the title compound.

¹ H-NMR (200 MHz, CDCl 3): δ 9.03 (s, 2H), 7.25 (s, 1OH), 5.05 (s, 4H)

3.92 (m, 2H), 3.20-2.90 (m, 4H), 1.78 (s, 6H), 1.44 (s, 6H)

IR (neat): 3388.3, 2936.0, 1701.6, 1418.9, 1277.5 cm ^-1

<Example 14>

(RR, SS) -4,8-diaza-3,6,6,9-tetramethylundecane-2,10-dione bisoxime

Preparation of (d.1-HMPAO)

(Example 14-1)

(RR, SS) -N, N-dibenzyloxycarbonyl-4,8-diaza-3,6,6,9-tetramethylundecane-2,10-dione bisoxime (2.3, obtained in Example 13) g, 4.25 mmol) is dissolved in 50 ml of ethanol and 10% palladium / carbon (900 mg) is carefully added. The reaction solution is stirred at room temperature under hydrogen stream for 2.5 hours. If no starting material is present on the TCL, the reaction solution is filtered through celite and the filtrate is concentrated. When the concentrated solution is about 2ml, add 3ml of ethyl acetate and stir for 10 minutes. Then, 6 ml of hexane was added thereto, stirred for 20 minutes, filtered, and the solid obtained was dried in vacuo to obtain 590 mg (51%) of the title compound.

¹ H-NMR (200 MHz, DMSO-d ₆ ): δ 10.27 (s, 2H), 3.10 (m, 2H), 2.O9 (m, 4H), 1.63 (s, 6H), 1.05 (d, 2H ), 0.77 (s, 6H)

IR (KBr): 3237.5, 2987.9, 2879.4, 1361.8, 837.2 cm ^-1

(Example 14-2)

(RR, SS) -N, N-dibenzyloxycarbonyl-4,8-diaza-3,6.6.9-tetramethylundecane-2,10-dione bisoxime (2.0 g, obtained in Example 13) 3.7 mmol) is dissolved in 50 ml of ethanol and 10% palladium / carbon (800 mg) and 1,4-cyclohexadiene (3.52 ml 37 mmol) are added slowly. The reaction solution is stirred at room temperature for 15 hours. When the starting material disappears on TLC, the reaction solution is passed through celite and the filtrate is concentrated. 3 ml of ethyl acetate was added to the concentrate, followed by stirring for 10 minutes. Then, 6 ml of hexane was added thereto, stirred for 20 minutes, filtered, and the obtained solid was dried in vacuo to yield 756 mg (75%) of the title compound.

In the present invention, two compounds having optical activity are used as starting materials, and the reaction selectivity is excellent, and the reaction conditions are mild, so that the manufacturing process is convenient, the yield of each step is relatively high, and the purity of the meso isomer is high (RR, SS) -4,8-diaza-3,6.6.9-tetramethylundecane-2,10-dione bisoxime derivatives can be prepared.

Claims (9)

To the 2,2-substituted malonic acid derivative of Formula 5, base and ethyl acylhalogen derivative selected from ethylchloroformate, isobutylchloroformate, t-butylchloroformate, and trimethylacetyl chloride were added to the reaction solution of Formula 6 -A hydrochloride salt of alanine methyl ester derivative or a hydrochloride salt of L-alanine methyl ester derivative of formula 7 is reacted to obtain a mixture of formulas 8a and 8b, and the resulting mixture of formulas 8a and 8b is reduced to Obtaining a mixture, introducing a protecting group into the mixture of formulas 9a and 9b to obtain a mixture of formulas 10a and 10b, and oxidizing the mixture of formulas 10a and 10b to obtain a mixture of formulas 11a and 11b, The mixture of 11a and 11b was reacted by adding Grignard reagent. A mixture of Formula 12a and Formula 12b was obtained, and the mixture of Formula 12a and Formula 12b was oxidized to obtain a mixture of Formula 13a and Formula 13b, and the mixture of Formula 13a and Formula 13b was added to a hydroxylamine derivative and an acid. Reacting to obtain a mixture of Formula 14a and Formula 14b, and then removing the protecting group from the mixture of Formula 14a and Formula 14b to prepare a mixture of Formula 15a and Formula 15b.

Wherein R is a lower alkyl group which may be arranged in a straight or branched chain having 1 to 6 carbon atoms, a cyclic type represented by-(CH2) n-, or-(CH2) aX- (CH2) b- It means the ring form which can be represented. In this case, X means N, O, and S, and n, a, and b are independent variables, respectively. R ₁ means a lower alkyl group which may be arranged in a straight or branched chain having 1 to 6 carbon atoms. R ₂ means a lower alkyl group, a lower alkene group or a lower alkane group which can be arranged in a straight or branched chain having 1 to 10 carbon atoms. R ₃ means H or a lower alkyl group or benzyl group which may be arranged in straight or branched chain of 1 to 6 carbon atoms, and P is methoxycarbonyl, 2,2,2-trichloroethoxycarbonyl, t A protecting group selected from -butyloxycarbonyl, allyloxycarbonyl, benzyloxycarbonyl, 4-nitrobenzyloxycarbonyl. delete delete delete delete delete delete delete delete