KR100739903B1 - Method of preparing tetrakis2-alkoxyisobutylisonitrilecopperItetrafluoro borate derivatives and new composition containing them for labelling agent of radioactive materials thereof - Google Patents

Abstract

The present invention relates to a method for preparing a tetrakis (2-alkoxyisobutylisonitrile) copper (I) tetrafluoroborate derivative represented by the following Chemical Formula 1 and to a radioactive material labeling composition comprising the compound, and a metalryl halide As a starting material, 2-alkoxyisobutylisonitrile was prepared through an alkoxylation step, azation step, amination step, and isonitrileation step, and the 2-alkoxyisobutylisonitrile was reacted with a copper compound to give tetra The present invention relates to a method for preparing a kiss (2-alkoxyisobutylisonitrile) copper (I) tetrafluoroborate derivative and a composition containing the same. It is possible to obtain a compound in a safe and high yield without using, and the radioactive material labeling composition of the present invention Zone and also is excellent in radioactive labeling efficiency.

Cu ((CH ₃ ) ₂ C (OR) CH ₂ NC) ₄ BF ₄

(Wherein R is as defined in the specification).

Description

Method for preparing tetrakis (2-alkoxyisobutylisonitrile) copper (I) tetrafluoroborate derivative and composition for labeling a new radioactive substance comprising the compound borate derivatives and new composition containing them for labeling agent of radioactive materials

The present invention relates to a method for preparing a tetrakis (2-alkoxyisobutylisonitrile) copper (I) tetrafluoroborate derivative represented by the following Chemical Formula 1, wherein the alkoxylation step and azide are performed using a metalyl halide as a starting material. 2-alkoxyisobutylisonitrile is prepared by a step of amination, amination and isonitrile, and the 2-alkoxyisobutylisonitrile is reacted with a copper compound to make tetrakis (2-alkoxyisobutylisonitrile) copper. (I) a method for producing a tetrafluoroborate derivative. The present invention also relates to a novel radioactive material labeling composition comprising the tetrakis (2-alkoxyisobutylisonitrile) copper (I) tetrafluoroborate derivative.

Formula 1                         

Cu ((CH ₃ ) ₂ C (OR) CH ₂ NC) ₄ BF ₄

(Wherein R is a C ₁ -C ₄ lower alkyl or branched alkyl group)

Radiopharmaceuticals have a structure capable of complexing with radioisotopes and are widely used for the diagnosis and treatment of diseases or for medical research.

Tetrakis (2-alkoxyisobutylisonitrile) copper (I) tetrafluoroborate compounds of the present invention are complexed with ^99m technetium (hereinafter referred to as " ^99m Tc") to include liposomes or vesicles and lipid membranes of a living body. It exhibits effective labeling properties for several cells and is used as an effective imaging agent for detecting abnormalities in tissues of various organs, especially the heart.

The tetrakis (2-alkoxyisobutylisonitrile) copper (I) tetrafluoroborate consists of a complex compound of 2-alkoxyisobutylisonitrile and copper, and the production method of 2-methoxyisobutylisonitrile is largely as follows. It can be summarized in three ways as follows.

1. A method developed by Bergstein et al. Is shown in Scheme 1 (European Patent No. 233,368).

As shown in Scheme 1, the synthesis of A and B using 2-amino-2-methyl-1-propanol and methylhydroxyisobutyrate as starting materials, respectively, consists of four and five steps, respectively. However, the synthesis method of A has a yield of 5.9%, and the synthesis method of B is very low at 8.1%. In particular, the "diphosgene" used in the last step is a poisonous substance that is harmful to the human body. Difficult to use

2. A method developed by Lamaringem et al. Is shown in Scheme 2 (US Pat. No. 4,864,051).                         

As shown in Scheme 2, the preparation method using 2-hydroxyisobutylonitrile as a starting material is composed of four steps and the total yield is 26%, which is relatively higher than the above method, but this method is also performed in the last step. There is a problem of using "diphosgene" harmful to the human body.

3. The method developed by Riteway et al. Is the same as Scheme 3 (US Pat. No. 5,210,270).

As shown in Scheme 3, the production method using isobutylene as a starting material is composed of four steps and the overall yield is 46%, which is very high compared with the existing method. However, isobutylene, which is used as a starting material, has a problem of being difficult to operate when used such as having to collect for a long time when reacting with a highly flammable gas.

Accordingly, the present inventors have tried to prepare 2-alkoxyisobutylisonitrile in a safe and high yield. As a result, as a starting material, the metal alkoxylation reaction step, the azidization step, the reduction step and the isonitrileation step are carried out. 2-alkoxyisobutylisonitrile was prepared in a yield, and the compound was reacted with a copper compound to prepare tetrakis (2-alkoxyisobutylisonitrile) copper (I) tetrafluoroborate derivative. The elimination of the use of toxic reagents and gases resulted in a safe and high yield manufacturing process. In addition, the new radioactive material labeling composition containing the compound was found to not reduce the radiolabeling efficiency even after long-term storage has completed the present invention.

It is an object of the present invention to prepare 2-alkoxyisobutylisonitrile in a relatively short stage of production process with excellent yield of reaction using metalyl halide as a starting material, and react a suitable metal complex to tetrakis (2-alkoxy). It is to provide a method for producing isobutyl isonitrile) copper (I) tetrafluoroborate derivative.                         

It is also an object of the present invention to provide a composition for labeling radioactive substances containing tetrakis (2-alkoxyisobutylisonitrile) copper (I) tetrafluoroborate derivatives, reducing agents and stabilizers.

The present invention to achieve the above object

1) alkoxylation by sequentially reacting the metalryl halide (2) with mercury acetate, sodium hydroxide, sodium borohydride in a suitable alcohol solvent as a starting material (step 1);

2) reacting the alkoxy compound (3) of step 1 with sodium azide in an appropriate solvent (step 2);

3) amination of the azide compound (4) of step 2 in the presence of lithium aluminum hydride (step 3);

4) isonitrile the amine compound (5) of step 3 using an inorganic base in the presence of chloroform and a phase transfer catalyst (step 4);

5) Reaction of the isonitrile compound (6) of step 4 with the copper compound (7) (step 5) of tetrakis (2-alkoxyisobutylisonitrile) copper (I) tetrafluoroborate derivative represented by the formula (1) It provides a manufacturing method.

In the above formula, R is a C ₁ -C ₄ lower alkyl group or a branched alkyl group, X is a halogen element, and R 'is a C ₁ -C ₄ lower alkyl group.

Preferably R is methyl, ethyl, propyl or butyl, X is chloride, bromine or iodine and R 'is ethyl or propyl.

In addition, the present invention,

1) alkoxylation by sequentially reacting the metalryl halide (2) with mercury acetate, sodium hydroxide, sodium borohydride in a suitable alcohol solvent as a starting material (step 1);

2) reacting the alkoxy compound (3) of step 1 with sodium azide in an appropriate solvent (step 2);

3) Amination of the azide compound (4) of step 2 with the amine compound (5) in the presence of lithium aluminum hydride (step 3);                     

4) Preparation of 2-alkoxyisobutylisonitrile represented by Chemical Formula 6 by isonitrile the amine compound (5) of step 3 using an inorganic base in the presence of chloroform and a phase transfer catalyst (step 4). Provide a method.

* (Wherein R, R 'and X are as defined above)

Hereinafter, the present invention will be described in each step.

Step 1: alkoxylation step

(Wherein R and X are as defined above).

In step 1, alkoxylated commercially available metalallyl halide (2) is alkoxylated through an oxyhydration-dehydration reaction under an alcohol solvent to prepare 2-alkoxyisobutyl halide (3).

Oxyhydration-dehydration reaction is a useful reaction that can introduce an alkoxy group to the double bond site, by reacting the metalyl halide (2) with mercury acetate, sodium hydroxide, sodium borohydride sequentially in a suitable alcohol solvent An alkoxy group is introduced.

In this case, the alkoxy group introduced into the 2-alkoxy isobutyl halide (3) may be changed according to the carbon number of the alcohol solvent used. In the present invention, an alcohol solvent including an alkyl group or a branched alkyl group of C ₁ -C ₄ is used. Preferably, one selected from methanol, ethanol, propanol or butanol is used.

Step 2: Azidation Step

(Wherein R and X are as defined above).

In step 2, the 2-alkoxyisobutyl halide (3) prepared in step 1 is subjected to nucleophilic substitution in a suitable solvent to azide to prepare 2-alkoxyisobutyl azide (4).                     

Solvents usable in step 2 include dimethyl sulfoxide, dimethylformamide, 1-methyl-2-pyrrolidinone, and the like, and are preferably reacted at 120 to 180 ° C. for 5 to 15 hours.

Step 3: Amination Step

(Wherein R is as defined above.)

In step 3, 2-alkoxyisobutyl azide (4) prepared in step 2 is prepared using lithium aluminum hydride to prepare 2-alkoxyisobutylamine (5).

At this time, the reaction is preferably reacted for 30 to 60 minutes at 0 ~ 5 ℃.

Step 4: isonitrilation step

(Wherein R is as defined above.)

In step 4, the 2-alkoxyisobutylamine (5) prepared in step 3 is subjected to isonitization by adding an inorganic base in the presence of chloroform and a phase transfer catalyst to form 2-alkoxyisobutylisonitrile (6). Manufacture.

In the present invention, the phase transfer catalyst is a tetraalkylammonium halide compound represented by R ″ ₄ NX, wherein R ″ is a C ₁ to C ₄ alkyl group and X is a halogen group element, preferably tetraalkylammonium chloride, tetra Alkyl ammonium bromide, tetraalkyl ammonium iodine or tetraalkyl ammonium hydroxide are used.

In the present invention, 2-alkoxyisobutylamine (5) is dissolved in chloroform, a suitable phase transfer catalyst and an inorganic base are added, and reacted for 1 to 3 hours at reflux temperature of the organic solvent. To prepare.

At this time, sodium hydroxide and potassium hydroxide are used as the inorganic base, and sodium hydroxide is preferably used.

Step 5: complex compound preparation

(Wherein R, X and R 'are as defined above).

In step 5, the 2-alkoxyisobutylisonitrile (6) prepared in step 4 is reacted with tetrakis (alkylnitrile) copper (I) tetrafluoroborate (7) in a suitable solvent to form tetrakis represented by the formula (1). 2-alkoxyisobutylisonitrile) copper (I) tetrafluoroborate derivative (1) was prepared.

Specifically, 0.2 to 0.3 equivalent, preferably 0.25 equivalent, of tetrakis (alkylnitrile) copper (I) tetrafluoroborate (7) to 1 equivalent of 2-alkoxyisobutylisonitrile (6) is added to 0-30. It is made to react at 10 degreeC for 10 to 60 minutes, and tetrakis (2-alkoxy isobutyl isonitrile) copper (I) tetrafluoro borate derivative (1) is manufactured.

Tetrakis (alkylnitrile) copper (I) tetrafluoroborate preferably uses tetrakis (acetonitrile) copper (I) tetrafluoroborate or tetrakis (propionitrile) copper (I) tetrafluoroborate do.

The solvent also uses an alcohol solvent, preferably methanol or ethanol.

As described above, the present invention provides 2-alkoxy isobutyl isonitrile (6) in a four-step manufacturing process having excellent yield of reaction and relatively short production time using metall halide (2) as a starting material, and suitable copper Reaction with compound (7) produces tetrakis (2-alkoxyisobutylisonitrile) copper (I) tetrafluoroborate derivative (1).

The present invention also provides a composition for labeling radioactive substances containing tetrakis (2-alkoxyisobutylisonitrile) copper (I) tetrafluoroborate derivative (1), a reducing agent and a stabilizer.

Tetrakis (2-alkoxyisobutylisonitrile) copper (I) tetrafluoroborate derivatives (1) prepared in the present invention can form complexes with radioactive materials, which is tetrakis (2-alkoxyisobutylisonitrile) It is supplied in the form of a composition containing a copper (I) tetrafluoroborate derivative (1), a reducing agent and a stabilizer and used in admixture with a radioactive substance or an aqueous solution thereof immediately before administration to a patient.

In order for the tetrakis (2-alkoxyisobutylisonitrile) copper (I) tetrafluoroborate derivative (1) to function for radiolabeling, a reducing agent is required. As the reducing agent, a reducing agent commonly used in this field may be added, and SnX ₂ is preferable. X is selected from halogen, acetate, sulfate, oxalate, or succinate, and preferably selected from tin chloride, tin acetate, tin oxalate and tin succinate.

In order for the tetrakis (2-alkoxyisobutylisonitrile) copper (I) tetrafluoroborate derivative (1) of the present invention to label radioactive substances, a tin (II) compound which is a reducing agent is required, and the tin (II) Since the compound oxidizes divalent tin to tetravalent tin over time, the reducing power is reduced, so that the label for the radioactive material of tetrakis (2-alkoxyisobutylisonitrile) copper (I) tetrafluoroborate derivative (1) Decreases efficiency [Tofe AJ, Francis MD. In vitro stabilization of a low-tin bone-imaging agent ( ^99m Tc-Sn-HEDP) by ascorbic acid. J. , Nucl ., Med ., 1976 ; 17 (9): 820-5] Stabilizers are added to increase the stability of the tin (II) compound. As a stabilizer, a stabilizer commonly used in this field may be ascorbic acid, cysteine, gentisic acid or ethanol, and preferably ascorbic acid. Ascorbic acid may play a role in maintaining high radiochemical labeling efficiency of radioactive material by inhibiting oxidation of divalent tin at low concentrations. The result is an extended shelf life of the diagnostic kits produced and an increased label stability upon labeling with radioactive material.

Tetrakis (2-alkoxyisobutylisonitrile) copper (I) tetrafluoroborate derivative (1) prepared in the present invention may be mixed with a reducing agent and a stabilizer, injected into a sterile vial, lyophilized, and then provided as a kit. .

The lyophilized diagnostic kit is labeled with a radioactive material, for example ^99m technetium to form a complex, the tetrakis (2-alkoxyisobutylisonitrile) copper (I) tetrafluoroborate derivative of the present invention (1 The ^99m technetium complex of) ^exhibits effective labeling properties for many cells, including liposomes or vesicles and lipid membranes in vivo, and can also be used as an effective imaging agent for detecting abnormalities in tissues of various organs, especially the heart. .

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.

The molecular structure was confirmed by infrared spectroscopy (IR), ultraviolet-visible light spectroscopy (UV-VIS), nuclear magnetic resonance spectroscopy (NMR), mass spectrometry (MA), and elemental analysis of representative compounds and comparison of actual values. The purity of was confirmed by HPLC analysis.

Preparation Example 1 Preparation of Tetrakis (propionitrile) copper (I) tetrafluoroborate

After dissolving Cu (BF ₄ ) ₂ .xH ₂ O (2.6 g, 11.0 mmol) in 6 ml of propionitrile, the temperature was raised to 97 ° C., the reflux temperature of propionitrile, and then cooled immediately. Copper powder was added to this semi-solution until the blue color of the reaction solution became faint, followed by stirring for 30 minutes, and then the reaction was terminated. The reaction solution was filtered, the filtrate was concentrated, ether was added to the residue, followed by solidification, followed by filtration and drying to obtain 3.9 g (96%) of the desired tetrakis (propionitrile) copper (I) tetrafluoroborate. Got.

Example 1 Preparation of Tetrakis (2-methoxyisobutylisonitrile) Copper (I) Tetrafluoroborate (Setamibi)

(Step 1): Preparation of 2-methoxyisobutyl chloride

Metallic chloride (20 g, 220.9 mmol) was slowly added dropwise to a solution of mercury acetate (74 g, 232.2 mmol) in 300 ml of methanol, followed by stirring at room temperature for 1 hour. Subsequently, the reaction solution was cooled to 0-5 ° C., and a solution of sodium hydroxide (21 g, 525 mmol) in 200 ml of methanol was slowly added dropwise to the reaction solution over 30 minutes, followed by stirring at room temperature for 1 hour. The reaction solution was cooled again to 0-5 ° C. and sodium borohydride (5.5 g, 145 mmol) was added several times. Then, the temperature of the reaction solution was raised to room temperature and stirred for 2 hours. After cooling the temperature of the reaction solution to 0 ~ 5 ℃ slowly added 300 ml of saturated citric acid aqueous solution to the reaction solution and extracted with ether. The organic layer was washed with saturated aqueous sodium bicarbonate solution and saturated brine, dehydrated with anhydrous magnesium sulfate (MgSO ₄ ), and then the filtrate was concentrated at a low temperature to give 25 g of 2-methoxyisobutyl chloride as a target compound (92.3%). )

¹ H-NMR (200 MHz, CDCl ₃ ): δ 1.23 (s, 6H), 3.22 (s, 3H), 3.45 (s, 2H)

IR (neat): 2959.5, 2820.1, 1415.1, 1359.9, 1064.8, 734.5 cm ^-1

(Step 2): Preparation of 2-methoxyisobutyl azide

2-methoxyisobutyl chloride (16.7 g, 136.3 mmol) obtained in the above step was dissolved in 120 ml of dimethyl sulfoxide, sodium azide (26.6 g, 409.2 mmol) was added, followed by stirring at 180 ° C. for 12 hours. . After completion of the reaction, the mixture was cooled, ether was added, washed with saturated citric acid solution, saturated sodium bicarbonate solution and saturated salt water in that order, and the organic layer was dehydrated with anhydrous magnesium sulfate. Subsequently, the filtrate was concentrated about 1/2 after filtration and used for the reaction as it was without any purification.

¹ H-NMR (200 MHz, CDCl ₃ ): δ 1.18 (s, 6H), 3.18 (s, 2H), 3.22 (s, 3H)

(Step 3): Preparation of 2-methoxyisobutylamine

LiAlH ₄ (10.3 g, 271.4 mmol) was added to the reactor, and 150 ml of ether was added thereto, followed by cooling to 0-5 ° C. To the 2-methoxyisobutyl azide obtained in the above step was slowly added dropwise and stirred for 30 minutes at 0 ~ 5 ℃. After the reaction was completed, 10.3 ml of water, 10.3 ml of 15% aqueous sodium hydroxide solution, and 30.9 ml of water were slowly added dropwise thereto, followed by vigorous stirring at room temperature for 20 minutes. After the reaction was completed, the resulting solid was filtered and washed with 50 ml of ether. The filtrate and washings were concentrated at a low ?? degree (0-5 ° C.) and the residue was then simply distilled at 120-125 ° C. to give 10.4 g (74.0%) of 2-methoxyisobutylamine.

¹ H-NMR (200 MHz, CDCl ₃ ): δ1.09 (s, 6H), 1.61 (s, 2H), 2.56 (s, 2H), 3.13 (s, 3H)

(Step 4): Preparation of 2-methoxyisobutylisonitrile

2-methoxyisobutylamine (20 g, 194 mmol) obtained in the above step was dissolved in 200 ml of chloroform, and then tetra-n-butylammonium bromide (6.25 g, 19.4 mmol) was added. A solution of sodium hydroxide (31 g, 775 mmol) in 100 ml of water was slowly added dropwise thereto, followed by stirring under reflux for 1 hour. After completion of the reaction, after cooling to room temperature, the resulting layer was separated, and the aqueous layer was reextracted with methylene chloride. After separation of the layers, the selected organic layer and the organic layer extracted from the aqueous layer were mixed, the mixture was washed with saturated brine, dehydrated with anhydrous magnesium sulfate and filtered. The filtrate was then concentrated under reduced pressure (25 mmHg, 55-60 ° C.) to give 13.7 g (62.5%) of the desired 2-methoxyisobutylisonitrile.

¹ H-NMR (200 MHz, CDCl ₃ ): δ 1.25 (s, 6H), 3.23 (s, 3H), 3.35 (t, 2H)

IR (neat): 2829.5, 2148.5, 1459.7, 1370.0 cm ^-1

(Step 5): Preparation of tetrakis (2-methoxyisobutylisonitrile) copper (I) tetrafluoroborate (cetamibi)

Tetrakis (propionitrile) copper (I) tetrafluoroborate (1.5 g, 4.05 mmol) obtained in Preparation Example 1 was dissolved in 20 ml of methanol, followed by 2-methoxyisobutylisonitrile (1.83). g, 16.2 mmol) was slowly added dropwise and stirred at room temperature for 1 hour. After the reaction was completed, ether was gradually added to the residue obtained by concentrating the reaction solution to give a solid. The resulting solid was filtered and dried to give 2.32 g (95.0%) of the desired tetrakis (2-methoxyisobutylisonitrile) copper (I) tetrafluoroborate.

¹ H-NMR (200 MHz, CDCl ₃ ): δ 1.25 (s, 6H), 3.22 (s, 3H), 3.57 (s, 2H)

¹³ C-NMR (200 MHz, CDCl ₃ ): δ 22.4, 49.9, 51.6, 73.2

Elemental analysis (C ₂₄ H ₄₄ N ₄ O ₄ CuBF ₄ ): Theoretical C: 47.806 H: 7.355 N: 9.292

Found C: 47.416 H: 7.219 N: 9.359

Example 2 Preparation of 2-methoxyisobutyl azide

As a starting material, intermediate 2-methoxyisobutyl azide was prepared using metalryl bromide.

(Step 1): Preparation of 2-methoxyisobutyl bromide

23.3 g (94.2%) of 2-methoxyisobutyl bromide was obtained by the same method as Step 1 of Example 1 using metalryl bromide (20 g, 148.1 mmol).                     

¹ H-NMR (200 MHz, CDCl ₃ ): δ 1.31 (s, 6H), 3.24 (s, 3H), 3.41 (s, 2H)

(Step 2): Preparation of 2-methoxyisobutylisonitrile

2-methoxyisobutyl bromide obtained in the above step was carried out in the same manner as in Step 2 of Example 1 to obtain 2-methoxyisobutyl azide.

(Step 3): Preparation of 2-methoxyisobutylamine

Scheme 14

2-methoxyisobutyl azide obtained in the above step was carried out in the same manner as in Step 3 of Example 1 to obtain 2-methoxyisobutylamine.

(Step 4): Preparation of 2-methoxyisobutylisonitrile

Scheme 15

2-methoxyisobutylamine obtained in the above step was carried out in the same manner as in Step 4 of Example 1, to obtain 2-methoxyisobutylisonitrile.

Example 3 Preparation of Lyophilized Kit

To prepare a kit containing the compound of the present invention was carried out as follows.

Tetrakis (2-methoxyisobutylisonitrile) copper (I) tetrafluoroborate of the present invention was injected into a vial and an amount of stannous chloride, L-cysteine, sodium citrate, and mannitol were added. Ascorbic acid (0 ㎍, 7.5 ㎍, 75 ㎍) was added thereto, dissolved in low dissolved oxygen distilled water, and nitrogen gas was injected to replace the inside of the vial with nitrogen, followed by freeze drying for 40 hours. Prepared kits.

The components of the kit are as follows.

Tetrakis (2-methoxyisobutylisonitrile) copper (I) tetrafluoroborate compound of the present invention ... 1.0 mg

Tin chloride Chloride: 0.075 mg ··················· 0.075 mg                     

L-cysteine 1.0 mg

Sodium citrate ... 2.6 mg

20 mg of mannitol ····················

Ascorbic acid: 0 μg, 7.5 μg, 75 μg

Experimental Example 1 By Addition of Ascorbic Acid ^99m Radiochemical Labeling Efficiency of Tc Sestamibi

Using a lyophilized kit prepared in Example 3 to label ^99m technetium to prepare a radiopharmaceutical and to measure the labeling efficiency.

2 mL (3 mCi) of radioactive sodium pertechnate was added to the lyophilized kit and reacted at 95-100 ° C. for 15 minutes to label ^99m technetium. At this time, to measure the radiochemical labeling efficiency, chromatography was performed using Whatman No 1 and Instant thin-layer chromatography-silicagel (ITLC-SG) as a stationary phase, and acetone and physiological saline as the mobile phase, and the results were confirmed by a TLC scanner. was confirmed (Table 1) does not affect the ascorbic acid radioactive labeling efficiency was added as a stabilizer.

^{Radiochemical} Labeling Efficiency of ^99m Tc ^Cestamibi by Addition of Ascorbic Acid Ascorbic Acid Content Vial can Radiochemical Labeling Efficiency (%) ^99m Tc-MIBI Unreacted ^99m TcO ₄ ^- Colloid Compositions of the Invention 0 μg 3 98.0 ± 0.0 0.0 ± 0.0 2.0 ± 0.1 7.5 μg 3 98.7 ± 0.6 0.0 ± 0.0 1.3 ± 0.1 75 μg 3 99.0 ± 0.0 0.0 ± 0.0 1.0 ± 0.0   The experimental results are mean ± standard deviation

Experimental Example 2 By Addition of Ascorbic Acid as Reducing Agent ^99m Radiochemical Labeling Efficiency of Tc Sestamibi

Using a lyophilized kit to which only ascorbic acid was added as a reducing agent, ^99m technetium was labeled to prepare a radiopharmaceutical, and the labeling efficiency was measured.

Tetrakis (2-methoxyisobutylisonitrile) copper (I) tetrafluoroborate of the present invention was injected into a vial and a certain amount of sodium citrate and mannitol were added. Ascorbic acid (0 ㎍, 7.5 ㎍, 75 ㎍) was added thereto, dissolved in low dissolved oxygen distilled water, and nitrogen gas was injected to replace the inside of the vial with nitrogen, followed by freeze drying for 40 hours. Prepared kits.

2 mL (3 mCi) of radioactive sodium pertechnate was added to the lyophilized kit and reacted at 95-100 ° C. for 15 minutes to label ^99m technetium. At this time, chromatography was performed using Whatman No 1 and Instant thin-layer chromatography (silicagel) (ITLC-SG) as a stationary phase and acetone and physiological saline as a mobile phase to measure radiochemical labeling efficiency. When using ascorbic acid as a reducing agent ( Table 2 ), the labeling efficiency was less than 1% and it was confirmed that there was no reducing activity at all.

^{Radiochemical} Labeling Efficiency of ^99m Tc ^Cestamibi with Ascorbic Acid as Reducing ^Agent Ascorbic Acid Content Vial can Radiochemical Labeling Efficiency (%) ^99m Tc-MIBI Unreacted ^99m TcO ₄ ^- Colloid Comparative composition 0 μg 3 0.0 ± 0.0 98.0 ± 0.3 2.0 ± 0.0 7.5 μg 3 0.7 ± 0.1 94.0 ± 0.6 5.3 ± 0.1 75 μg 3 0.0 ± 0.0 96.5 ± 0.4 3.5 ± 0.1   The experimental results are mean ± standard deviation

Experimental Example 3 By Addition of Ascorbic Acid ^99m Stability of Accelerated Test of Tc Sestamibi

In order to determine the stability during the accelerated test of ^99m Tc ^sestamibi containing the compound of the present invention was carried out as follows.

The ^99m Tc ^sestamibi prepared in Experimental Example 2 was ^stored under accelerated test conditions of 40 ℃ ± 2 ℃, relative humidity 75 ± 5%. In order to measure the activity of ascorbic acid added as a stabilizer at 6 months after the accelerated test, the content of tin tin chloride was measured and shown in Table 3 below. As a result, it was confirmed that the content of Sn ⁺² having reducing activity was preserved 2 to 3 times of the content of tin chloride when ascorbic acid was added. As a result, the addition of ascorbic acid was evaluated to increase the stability of the vial by preventing the oxidation of stannous chloride. In addition, the radiolabeling efficiency showed a similar effect as the onset. In addition, the radiolabeling efficiency was measured 9 months after the accelerated test and is shown in Table 4 . The above conditions are expected to be about 4 times the long-term preservation test for the room temperature preservation drug. Ascorbic acid was added as a result of measuring the radiochemical labeling efficiency of ^99m Tc-sestamibi conducted up to 9 months after the accelerated test. In one case, the labeling efficiency increased by 5-8 times.

 Stability of TiN Chloride as a Reducing Agent by Addition of Ascorbic Acid Ascorbic Acid Content Vial can Tin chloride content (Sn (I),%) Compositions of the Invention 0 μg 3 13.4 ± 0.54 7.5 μg 3 41.8 ± 0.92 75 μg 3 30.8 ± 0.84   The experimental results are mean ± standard deviation

^{Radiochemical} Labeling Efficiency of ^99m Tc ^Sestamibi Ascorbic Acid Content Vial can Radiochemical Labeling Efficiency (%) ^99m Tc-MIBI Unreacted ^99m TcO ₄ ^- Colloid Compositions of the Invention 0 μg 2 10.0 70.0 20.0 7.5 μg 2 58.6 41.5 0.0 75 μg 2 92.0 0.0 8.0   The experimental results are mean ± standard deviation

According to Table 4 , it was confirmed that the radiochemical labeling efficiency can be extended to show a labeling efficiency similar to that of the first manufacturing ( Table 1 ), and the above results are chlorinated in order to increase the stability of the cestamabi formulation. It was found that addition of ascorbic acid is preferable as a stabilizer for preventing oxidation of tin tin.

Therefore, as described above, the manufacturing method of the present invention does not use diphosgene, which is harmful to the human body, compared to the conventional manufacturing method, and thus, tetrakis (2-alkoxyisobutylisonitrile) copper (I) tetra is safe and high in yield. Fluoroborate derivatives can be obtained, and the reaction step will be shortened to increase industrial value. In addition, the radiolabeling composition consisting of the compound, the reducing agent and the stabilizer of the present invention can be prepared as a lyophilized kit and excellent in radiolabeling efficiency even after long-term storage.

Claims (17)

1) alkoxylation by sequentially reacting metall halide (2) with mercury acetate, sodium hydroxide and sodium borohydride in a C ₁ -C ₄ lower or branched alcohol solvent (step 1 ); 2) reacting the alkoxy compound (3) of step 1 with sodium azide to azide (step 2); 3) reducing the azide compound (4) of step 2 to an amine group in the presence of lithium aluminum hydride (step 3); 4) isonitrile the amine compound (5) of step 3 using an inorganic base in the presence of chloroform and a phase transfer catalyst (step 4); 5) Tetrakis (2-alkoxyisobutylisonitrile) copper represented by the formula (1), characterized in that the step (4) is prepared by reacting the isonitrile compound (6) of step 4 with a copper compound (7). (I) A method for producing a tetrafluoroborate derivative. Scheme 4

In the above formula, R is a C ₁ -C ₄ lower alkyl group or a branched alkyl group, X is a halogen element, and R 'is a C ₁ -C ₄ lower alkyl group. delete 2. The tetrakis (2-alkoxyisobutylisonitrile) copper (I) tetrafluor according to claim 1, wherein the lower or branched alcohol of C ₁ -C ₄ of step 1 is methanol, ethanol, propanol or butanol. Process for the preparation of roborate derivatives. The catalyst of claim 1, wherein the phase transfer catalyst of step 4 is represented by the structural formula of R ″ ₄ NX, wherein R ″ is an alkyl group of C ₁ to C ₄ and X is an element of halogen group. Method for producing alkoxy isobutyl isonitrile) copper (I) tetrafluoroborate derivative. The tetrakis (2-alkoxy) of claim 1, wherein the phase transfer catalyst of step 4 is selected from tetraalkylammonium chloride, tetraalkylammonium bromide, tetraalkylammonium iodine or tetraalkylammonium hydroxide. Method for preparing isobutylisonitrile) copper (I) tetrafluoroborate derivatives. The method for preparing tetrakis (2-alkoxyisobutylisonitrile) copper (I) tetrafluoroborate derivatives according to claim 1, wherein the inorganic base of step 4 is selected from sodium hydroxide and potassium hydroxide. The method of claim 1, wherein step 5 is carried out by adding 0.25 equivalent of tetrakis (alkylnitrile) copper (I) tetrafluoroborate to 1 equivalent of 2-alkoxyisobutylisonitrile and reacting at 0 to 30 ° C for 10 to 60 minutes. A process for producing a tetrakis (2-alkoxyisobutylisonitrile) copper (I) tetrafluoroborate derivative, which is prepared. delete delete delete delete delete delete delete delete delete delete