KR101426872B1 - Heterocyclic derivatives or pharmaceutically acceptable salt thereof for metal isotope labeling and preparation method thereof - Google Patents

Abstract

The present invention relates to a heterocyclic derivative for a metal isotope label or a pharmaceutically acceptable salt thereof and a process for preparing the same, wherein the heterocyclic derivative according to the present invention or a pharmaceutically acceptable salt thereof is a compound having a heterocyclic structure Serves to fix the structure of the compound itself, thereby enhancing the coordination effect with the metallic isotope ruthecium, increasing the labeling efficiency of the metal isotope, and raising the stability. Therefore, the compound according to the present invention is introduced into various antibodies, It can be used effectively.

Description

[0001] The present invention relates to a heterocyclic derivative or a pharmaceutically acceptable salt thereof for metal isotope labeling and a method for preparing the same,

The present invention relates to a heterocyclic derivative for a metal isotope label, or a pharmaceutically acceptable salt thereof, and a process for producing the same.

Radioimmunotherapy (RIT) is a method of treatment using drugs with radioactive isotopes attached to antibodies. The antibody acts as a carrier to reach a specific cancer cell and the cytotoxicity of a radioactive isotope plays a role of killing cancer cells. The radioimmunotherapeutic agent used in this radioimmunotherapy is a drug consisting of zevaline (anti-CD 20 (antibody), 1B4M-DTPA (bifunctional chelating agent), ⁹⁰ Y (metal isotope) ), And many studies are under way.

Although the above-mentioned radioimmunotherapy technology has the advantage of selectively treating cancer cells without performing any surgical operation, research progress and results are progressing rather slowly. This is due to the fact that currently feasible chelating chelator is DOTA (1,4,7,10-tetraazacyclododecane-N, N ', N' '' - tetracetic acid) and DTPA (Diethylenetriamine pentaacetic acid) .

Since the DOTA-based compounds that can be used as the above-mentioned biologically active chelator compounds have a cyclic structure, there is a disadvantage in that the labeling reaction must proceed at a high temperature although stability and stability in vivo after labeling with metal is excellent. These disadvantages also lead to the problem that labeling of a protein such as an antibody is difficult, and a multi-step reaction according to a label is required. In addition, the compounds of the DTPA series have a disadvantage in that the labeling of the metal isotopes is easy, but the stability after labeling is poor.

Therefore, development of biologically active chelator compounds that can be used for radioimmunotherapy with high labeling efficiency and stability is urgently needed.

In order to overcome such disadvantages, the inventors of the present invention have studied a biocompatible chelator compound that appropriately compensates for a cyclic structure and a non-cyclic structure, and found that the non-cyclic amino acid (iminodiacetic acid) A chelator compound having heterocyclic group introduced therein was developed. In addition, it has been confirmed that the above heterocyclic derivative has good metal isotope labeling efficiency and stability, and the present invention has been completed.

It is an object of the present invention to provide a heterocyclic derivative or a pharmaceutically acceptable salt thereof.

It is another object of the present invention to provide a process for preparing the above heterocyclic derivative or a pharmaceutically acceptable salt thereof.

It is another object of the present invention to provide a heterocyclic derivative and a metal complex.

Another object of the present invention is to provide a medicament for radioimmunotherapy comprising the above heterocyclic derivative and a metal complex as an active ingredient.

It is still another object of the present invention to provide a kit for forming the complex with the heterocyclic derivative and the metal.

In order to achieve the above object, the present invention provides a heterocyclic derivative or a pharmaceutically acceptable salt thereof.

The present invention also provides a process for preparing the above heterocyclic derivative or a pharmaceutically acceptable salt thereof.

Further, the present invention provides the heterocyclic derivative and the complex of the metal.

The present invention also provides a medicament for radioimmunotherapy comprising the above heterocyclic derivative and a metal complex as an active ingredient.

Further, the present invention provides a kit for forming a complex with the heterocyclic derivative and the metal.

The heterocyclic compound according to the present invention or a pharmaceutically acceptable salt thereof may have a structure in which the heterocyclic structure fixes the structure of the compound itself, thereby enhancing the coordination effect with the metallic isotope rutheium, etc., Efficiency and stability are increased. Therefore, the compound according to the present invention can be effectively used for radioimmunotherapy by introducing it into various antibodies.

1 is a diagram showing the results of ITLC of a compound of Example 1 according to an embodiment of the present invention in a 75% methanol developing solvent.
FIG. 2 is a graph showing the results of ITLC on 75% methanol eluting solvent after labeling the compound of Example 1 according to an embodiment of the present invention with rutile.
FIG. 3 is a graph showing the results of ITLC of labeling rutheium on the compound of Example 2 according to an embodiment of the present invention, followed by 75% methanol elution solvent.
4a, 4b and 4c are molar ratios showing ruthenium labeling by bioadhesion with different ratios of the compound of Example 2 and the antibody according to an embodiment of the present invention.
FIG. 5 is a graph showing SDS-PAGE results of bioadhesion of a compound of Example 2 and an antibody at a ratio of 5: 1 according to an embodiment of the present invention. FIG.
FIG. 6 is a graph showing the results of labeling rutheum after bioadhesion of a compound of Example 2 and an antibody according to an embodiment of the present invention. FIG.

Hereinafter, the present invention will be described in detail.

The present invention provides a heterocyclic derivative represented by the following formula (I) or a pharmaceutically acceptable salt thereof.

In Formula 1,

,

,

또는

이고;R is

,

,

or

ego;

X is an acetyl group, a C ₁ -C ₄ aminoalkyl group;

n is an integer of 1 to 3;

Preferably,

또는

이고;Wherein R is

or

ego;

X is an acetyl group;

n is 1 or 2;

The heterocyclic derivatives represented by the formula (1) of the present invention can be used in the form of pharmaceutically acceptable salts, and acid addition salts formed by pharmaceutically acceptable free acids are useful as salts. The term pharmaceutically acceptable salt means a concentration that is relatively non-toxic to a patient and has a beneficial effect that is harmless to the patient, such that the side effects caused by the salt are any organic or inorganic salt of the base compound of Formula 1 that does not impair the beneficial effects of the base compound of Formula An inorganic addition salt. Examples of the inorganic acid include hydrochloric acid, bromic acid, nitric acid, sulfuric acid, perchloric acid and phosphoric acid. Examples of the organic acid include citric acid, acetic acid, lactic acid, maleic acid, (D) or (L) malic acid, maleic acid, methanesulfonic acid, ethanesulfonic acid, maleic acid, succinic acid, tartaric acid, galacturonic acid, embic acid, glutamic acid, aspartic acid, oxalic acid, P-toluenesulfonic acid, salicylic acid, citric acid, benzoic acid or malonic acid. These salts also include alkali metal salts (sodium salts, potassium salts, etc.) and alkaline earth metal salts (calcium salts, magnesium salts, etc.). For example, the acid addition salt may be selected from the group consisting of acetate, aspartate, benzoate, besylate, bicarbonate / carbonate, bisulfate / sulfate, borate, camylate, citrate, eddylate, Hydrobromide / bromide, hydroiodide / iodide, isethionate, lactate, malate, malate, glucoside, gluconate, gluconate, glucuronate, glucuronate, hexafluorophosphate, hibenzate, hydrochloride / Hydrogen phosphate, dihydrogen phosphate, dihydrogen phosphate, dihydrogen phosphate, dihydrogen phosphate, dihydrogen phosphate, dihydrogen phosphate, dihydroxyacetate, Lactate, stearate, succinate, tartrate, tosylate, trifluoroacetate Diethylamine, diolamine, glycine, lysine, magnesium, meglumine, olamine, potassium, sodium, tromethamine, zinc salts, and the like. Preferred is hydrochloride or trifluoroacetate.

The heterocyclic derivatives of formula (1) of the present invention include not only pharmaceutically acceptable salts, but also all salts, isomers, hydrates and solvates which can be prepared by conventional methods.

The addition salt according to the present invention can be prepared by a conventional method, for example, by dissolving the compound of Chemical Formula 1 in a water-miscible organic solvent such as acetone, methanol, ethanol, acetonitrile, etc., Followed by precipitation or crystallization. Subsequently, in this mixture, a solvent or an excess acid is evaporated and dried to obtain an additional salt, or the precipitated salt may be produced by suction filtration.

The present invention also relates to a process for producing a compound represented by the formula (1)

Reacting the compound of formula (2) as a starting material in a base and an organic solvent to prepare a compound of formula (3) (step 1);

Reacting the compound of Formula 3 prepared in Step 1 with ammonia in an organic solvent to prepare a compound of Formula 4 (Step 2);

Reacting the compound of Formula 4 prepared in Step 2 with an organic solvent to prepare a compound of Formula 5 (Step 3);

Reacting the compound of Formula 5 prepared in Step 3 with a compound of Formula 6 under a base to prepare a compound of Formula 7 (Step 4);

Reacting the compound of Formula 7 prepared in Step 4 with sodium salt in an organic solvent to prepare a compound of Formula 8 (Step 5);

Reacting the compound of Formula 8 prepared in Step 5 with a compound of Formula 9 in an organic solvent to prepare a compound of Formula 10 (Step 6);

Reacting the compound of formula 10 prepared in step 6 with an acid to produce a compound of formula 11 (step 7);

Reacting the compound of formula (11) prepared in step 7 with a compound of formula (12) in an organic solvent to prepare a compound of formula (13) (step 8);

Reacting the compound of formula 13 prepared in step 8 in an organic solvent to prepare a compound of formula 14 (step 9); And

The present invention provides a process for preparing a compound represented by the formula (1), which comprises: (1) reacting the compound of the formula (14) prepared in the step (9) with an acid in an organic solvent to prepare a compound of the formula

[Reaction Scheme 1]

.

.

(Wherein R, X and n are as defined in the above formula (1), R ¹ is a derivative of R, and R ² is a linear or branched alkyl of C ₁ to C ₅ )

Hereinafter, the manufacturing method will be described step by step.

Step 1

Step 1 according to the present invention is a step of preparing a compound of Formula 3 by reacting the compound of Formula 2 as a starting material in a base and an organic solvent.

The base used in step 1 may be selected from triethylamine, pyridine, dimethylaminopyridine and the like, preferably triethylamine.

The organic solvent is preferably at least one solvent selected from the group consisting of ether, ethyl acetate, methylene chloride, tetrahydrofuran and methanol, and more preferably ether and methanol.

The reaction temperature of step 1 is preferably from room temperature to 50 ° C, and the reaction time is more preferably from 8 to 15 hours.

Step 2

Step 2 according to the present invention is a step of reacting the compound of Formula 3 prepared in Step 1 with ammonia in an organic solvent to prepare a compound of Formula 4. [

The organic solvent used in step 2 may be selected from methanol, ethyl acetate, tetrahydrofuran, N, N-dimethylformamide and the like, preferably methanol.

The reaction of step 2 is preferably carried out at a temperature of from room temperature to 50 ° C for 40 to 55 hours, more preferably at room temperature for 45 to 50 hours.

Step 3

Step 3 according to the present invention is a step of reacting the compound of Formula 4 prepared in Step 2 in an organic solvent to prepare a compound of Formula 5. [

The organic solvent used in step 3 may be selected from tetrahydrofuran, methylene chloride, acetone and the like, preferably tetrahydrofuran.

The reaction of step 3 is preferably carried out at 60 to 80 ° C for 17 to 30 hours, more preferably at 65 to 75 ° C for 20 to 25 hours.

Step 4

Step 4 according to the present invention is a step of reacting the compound of Formula 5 prepared in Step 3 with a compound of Formula 6 under a base to prepare a compound of Formula 7.

The base used in step 4 may be selected from among pyridine, triethylamine, dimethylaminopyridine and the like, preferably pyridine.

The reaction of step 4 is preferably carried out at 40 to 60 ° C for 1 to 5 hours, more preferably at 45 to 55 ° C for 1 to 3 hours.

Step 5

Step 5 according to the present invention is a step of reacting the compound of formula (7) prepared in step 4 with sodium salt in an organic solvent to prepare a compound of formula (8).

The organic solvent used in step 5 may be selected from methanol, ethanol, acetone, ethyl acetate and the like, preferably methanol.

The reaction of step 5 is preferably carried out at room temperature for 1 to 5 hours, more preferably for 2 to 4 hours.

Step 6

The step 6 according to the present invention is a step of reacting the compound of the formula 8 prepared in the step 5 with the compound of the formula 9 in an organic solvent to prepare the compound of the formula 10.

The organic solvent used in step 6 may be selected from N, N-dimethylformamide, toluene, methanol, acetone, ethyl acetate and the like, preferably N, N-dimethylformamide.

The reaction of step 6 is preferably carried out at room temperature for 20 to 28 hours, more preferably for 24 hours.

Step 7

Step 7 according to the present invention is a step of reacting the compound of Formula 10 prepared in Step 6 with an acid to prepare a compound of Formula 11.

The acid used in step 7 may be selected from sulfuric acid, hydrochloric acid, nitric acid and the like, preferably sulfuric acid.

The reaction of Step 7 is preferably carried out at 100 to 150 ° C for 1 to 5 hours, more preferably at 110 to 130 ° C for 1 to 3 hours.

Step 8

Step 8 according to the present invention is a step of reacting the compound of Formula 11 prepared in Step 7 with the compound of Formula 12 in an organic solvent to prepare the compound of Formula 13.

The organic solvent used in step 8 may be selected from N, N-dimethylformamide, ethyl acetate, tetrahydrofuran, methylene chloride and the like, preferably N, N-dimethylformamide.

The reaction of step 8 is preferably carried out at 20 to 70 ° C for 40 to 60 hours, more preferably at 30 to 50 ° C for 45 to 50 hours.

Step 9

Step 9 according to the present invention is a step of preparing a compound of Formula 14 by reacting the compound of Formula 13 prepared in Step 8 in an organic solvent.

The organic solvent used in step 9 may be selected from methanol, ethanol, acetone, ethyl acetate and the like, preferably methanol.

The reaction of step 9 is preferably carried out at room temperature for 15 to 30 hours, more preferably for 20 to 30 hours.

Step 10

Step 10 according to the present invention is a step of preparing the compound of Formula 1 by reacting the compound of Formula 14 prepared in Step 9 with an acid and an organic solvent.

The organic solvent used in step 10 may be selected from methylene chloride, ethyl acetate, acetone, acetonitrile and the like, preferably methylene chloride.

The reaction of Step 10 is preferably carried out at 20 to 50 ° C for 30 minutes to 4 hours, more preferably at room temperature to 40 ° C for 30 minutes to 2 hours.

Further, the present invention provides a heterocyclic derivative and a complex of a metal represented by the formula (1). In this case, the metal is preferably a metal isotope, and more preferably is lutetium (Lu-177).

The present invention also provides a medicament for radioimmunotherapy comprising the heterocyclic derivative and the metal complex of Formula 1 as an active ingredient.

The heterocyclic compound has a heterocyclic structure that fixes the structure of the compound itself, thereby enhancing the coordination effect with the metal isotope ruthemium. Therefore, the heterocyclic derivative according to the present invention can be usefully used as a medicament for radioimmunotherapy by introducing it into various antibodies.

Hereinafter, the present invention will be described in more detail with reference to examples. However, the following examples are illustrative of the present invention, and the contents of the present invention are not limited by the following examples.

< Example  (S) -2,2 ', 2' ', 2' '- (((2- (4- Aminobenzyl ) -1,4- Diazepan -1,4- Dill ) Bis (ethane-2,1- Dill )) Bis ( Azanetree )) Tetraacetic acid (1a)

Step 1: (S) - (+) - 4- Nitrophenylalanine Methyl ester  Produce

(S) - (+) - 4-nitrophenylalanine methyl ester hydrochloride (10 g, 38.4 mmol) were added to 5 ml of methanol and stirred at 40 ° C for 20 minutes. After adding 10 ml of triethylamine, 200 ml of ether was added and the mixture was stirred at room temperature for one day. At this time, a precipitate of triethylamine hydrochloride was formed. The precipitate was filtered through a reduced pressure filter, and 2 ml of methanol and 5 ml of triethylamine were added to the precipitate, and the mixture was stirred again in 100 ml of ether for 12 hours. After the precipitate was removed by vacuum filtration, the obtained solution was combined with the previously obtained solution and concentrated under reduced pressure. 200 ml of ether was added to the concentrated solution to remove residual triethylamine hydrochloride precipitate. The concentrate was distilled under reduced pressure to obtain the target compound (S) - (+) - 4-nitrophenylalanine methyl ester as a clear yellow sticky liquid (Yield: 100%, 8.6 g).

Step 2: (S) - (+) - 4- Nitrophenylalanine  Preparation of amide

(+) - 4-nitro-phenylalanine was added to 70 ml of 7N NH _{3 ·} MeOH under ester 0 ℃ condition was removed (8.6 g) for all of the remaining solution was reduced to prevent the rubber stopper (S) obtained in the above Step 1 The mixture was sealed and stirred for 1 hour. The reaction was then continued at room temperature for 48 hours. Over time, a yellow precipitate was formed and the completion of the reaction was confirmed by TLC (MC: MeOH = 9: 1, Rf = 0.3). The product solution was distilled under reduced pressure to remove the solvent and ammonia to obtain the target compound as a yellow solid (yield: 98%, 7.86 g).

Step 3: 1- (p- Nitrobenzyl ) Ethylenediamine  Produce

7.86 g of the (S) - (+) - 4-nitrophenylalaninamide obtained in the above step 2 was dissolved in 40 ml of THF. 100 ml of 1 M borane / THF solution was added under nitrogen atmosphere at 0 ° C, and the reaction was continued for 24 hours under reflux at 70 ° C. TLC was confirmed to confirm the removal of the reactant. In the presence of the unreacted reactant, 20 ml of a 1 M borane / THF solution was added to proceed the reaction. After completion of the reaction, 1 drop of distilled water corresponding to about 10 ml was added in an ice bath to make the borane inactive. The THF solution was distilled under reduced pressure until the solution became 1/3, 10 ml of 6 M HCl solution was added dropwise on an ice bath, and the mixture was stirred for 24 hours. Ammonia water was added to adjust the pH to 12 or more, followed by stirring for about 1 hour. The organic material was extracted with dichloromethane and distilled under reduced pressure to obtain 1- (p-nitrobenzyl) ethylenediamine as a target compound in the form of a viscous sticky liquid (yield: 93%, 6.82 g).

Step 4: Preparation of N, N ' - Ditosyl -1- (p- Nitrobenzyl ) Ethylenediamine  Produce

6.82 g of 1- (p-nitrobenzyl) ethylenediamine obtained in the above Step 3 was dissolved in 50 ml of pyridine, 2.4 equivalents (16 g) of tosyl chloride was added to 30 ml of pyridine After dissolving, small amounts were added. After reaction at 50 ° C for 2 hours, the reaction was confirmed by TLC (ethyl acetate: hexane = 1: 2). When the product was present, tosyl chloride was further added and the reaction was further continued for 1 hour. After the completion of the reaction was confirmed, the pyridine was distilled under reduced pressure to remove the solvent, and then the residual pyridine was removed using toluene. The organic material was extracted with dichloromethane and distilled water and distilled under reduced pressure. The resulting black sticky liquid was subjected to primary column separation under dichloromethane: methanol (95: 5). The resulting mixture was further purified twice using a solution of ethyl acetate: hexane (1: 2) to obtain the desired compound N, N'-ditosyl-1- (p-nitrobenzyl) ethylenediamine as a pale yellow solid (Yield: 83%, 14.6 g).

Step 5: N, N ' - Ditosyl -1- (p- Nitrobenzyl ) Ethylenediamine Disodium  Manufacture of salt

10 g of the N, N'-ditosyl-1- (p-nitrobenzyl) ethylenediamine prepared in the step 4 was dissolved in 50 ml of ethanol. 2 equivalents (0.91 g) of sodium was dissolved in 50 ml of ethanol and added to N, N'-ditosyl-1- (p-nitrobenzyl) ethylenediamine dissolved in ethanol at 0 ° C. After stirring for 10 minutes, the reaction was carried out under reflux conditions for 3 hours. Over time, the solution gradually became black and a yellow precipitate formed. The solution was distilled under reduced pressure and reduced to 1/4 and stored in a refrigerator for one day to form a precipitate. The precipitate was filtered under reduced pressure to obtain N, N'-ditosyl-1- (p-nitrobenzyl) ethylenediamine disodium salt (Yield: 55%, 6 g).

Step 6: (S) -2- (4- Nitrobenzyl ) -1,4- Ditosyl -1,4- Diazepan  Produce

Nitrogen atmosphere 5 g of the compound N, N'-ditosyl-1- (p-nitrobenzyl) ethylenediamine disodium salt obtained in Step 5 was dissolved in 200 ml of dimethylformamide (DMF) at 0 ° C. After complete dissolution, 1.2 equivalents (2.21 g, 1.12 ml) of 1,3-dibromopropane was added dropwise over 20 minutes. After the reaction was carried out at room temperature for 24 hours, the presence of the reaction product was confirmed by using TLC (ethyl acetate: hexane = 1: 2, Rf = 0.3) and adding 0.3 equivalent of 1,3-dibromopropane Respectively. After completion of the reaction, dimethylformamide was distilled off under reduced pressure and dissolved in ethyl acetate. The reaction product by-product, NaBr, was removed by filtration and distilled under reduced pressure to remove the mobile phase solution (Ethyl acetate: hexane = 1: 2) to give the target compound as a pale yellow powder (yield: 57%).

Step 7: (S) -2- (4- Nitrobenzyl ) -1,4- Diazepan  Produce

2 g of the (S) -2- (4-nitrobenzyl) -1,4-ditosyl-1,4-diazine plate prepared in the above step 6 was added to a 25 ml 1-neck round bottom flask, the cH ₂ SO ₄ 5 ml were added. At this time, it was confirmed that the color changed to black, and the reaction solution was stirred at 130 ° C for about 2 hours to remove the tosyl group. 150 ml of ether was added to a 250 ml round-bottomed flask, and 1 drop of the reaction solution was added thereto using a pastel pipette in an ice bath. After standing for more than 1 hour to form a precipitate, the supernatant was transferred to another beaker and stored. The above procedure was repeated one more time and 3N NaOH was added to the obtained black oil to adjust pH = 11, and the product was extracted with methylene chloride. The resulting organic layer was distilled under reduced pressure to obtain (S) -2- (4-nitrobenzyl) -1,4-diazepane (yield: 62%) as a yellow oil-like target compound.

Step 8: (S) - Tetra -t-butyl 2,2 ', 2 ", 2' '- (((2- (4- Nitrobenzyl ) -1,4- Diazepan -1,4- Dill ) Bis (Ethane-2,1- Dill )) Bis ( Azanetree )) Tetraacetate  Produce

0.54 g of the (S) -2- (4-nitrobenzyl) -1,4-diazepane obtained in the above step 7 was dissolved in 20 ml of DMF at room temperature under a nitrogen atmosphere, and then diisopropylethylamine (DIPEA, 10 eq , 2.98 g) and potassium iodide (KI, 1.2 eq, 0.46 g). Di-t-butyl 2,2 '- ((2-bromoethyl) azandiyl) diacetate was dissolved in 20 ml of DMF, and the mixture was stirred for 30 minutes using a dropping funnel. Lt; / RTI &gt; Thereafter, the temperature was raised to 40 ° C and the reaction was allowed to proceed for 2 days. After completion of the reaction, DMF was removed, dichloromethane was added to filter out precipitate which was a by-product of the reaction, and the filtrate was distilled under reduced pressure to obtain a product, which was subjected to column separation using methylene chloride: methanol = 9: 1 to obtain (S) Bis (ethane-2,1-diyl) bis (2,2 ', 2 &quot; ) Bis (azanetriyl)) tetraacetate. The color of the obtained compound was a pale yellow viscous liquid and the yield was 60%.

Step 9: (S) - Tetra -t-butyl 2,2 ', 2 ", 2' '- (((2- (4- Aminobenzyl ) -1,4- Diaz Plate-1, 4- Dill ) Bis (Ethane-2,1- Dill )) Bis ( Azanetree )) Tetraacetate  Produce

(S) -tetra-t-butyl 2,2 ', 2 ", 2''- (((2- (4-nitrobenzyl) -1,4- Bis (ethane-2,1-diyl) bis (azanetriyl)) tetraacetate was dissolved in 20 ml of methanol at room temperature, and 0.1 g of 10% Pd / C was added thereto. . After the hydrogen atmosphere was established, the reaction was carried out for 24 hours. The completion of the reaction was confirmed by using TLC (methylene chloride: methanol = 9: 1, Rf = 0, 25). Pd / C and residual impurities were removed using Celite and the filtrate was distilled under reduced pressure to obtain the target compound (S) -tetra-t-butyl 2,2 ', 2 ", 2''' - (Ethane-2,1-diyl)) bis (azanetriyl)) tetraacetate was obtained as a pale yellow viscous liquid And the following procedure was carried out without separate separation and purification.

Step 10: (S) -2,2 ', 2 ", 2' '- (((2- (4- Aminobenzyl ) -1,4- Diazepan -1,4- Dill ) Bis (Ethane-2,1- Dill )) Bis ( Azanetree )) Tetraacetic acid (1a)

(S) -tetra-t-butyl 2,2 ', 2'',2''' - (((2- (4-aminobenzyl) Bis (ethane-2,1-diyl) bis (azanetriyl)) tetraacetate (0.72 g) was dissolved in 20 ml of dichloromethane at room temperature, 10 ml of trifluoroacetic acid (TFA) And the mixture was stirred at room temperature for 30 minutes. The reaction temperature was raised to 40 ° C and further stirring was performed for 2 hours to remove the t-butyl group. After completion of the reaction, the solvent was distilled off under reduced pressure, 20 ml of dichloromethane was added, and the remaining TFA was removed by distillation under reduced pressure. The resulting compound was dissolved in a small amount of methanol, and then ether was added to perform recrystallization. As a result of confirming using Ms, a value of 524.3 having a molecular weight + 1 peak was obtained in a positive mode. The recrystallized compound was obtained in the form of a pale yellow powder.

< Example  2 (S) -2,2 ', 2 ", 2' '- ((2- (4- Isothiocyanatobenzyl ) -1,4- Diazepan -1,4-diyl) Bis (Ethane-2,1- Dill )) Bis ( Azanetree )) Tetraacetic acid (1b)

(S) -2,2 ', 2 ", 2''' - (((2- (4-aminobenzyl) -1,4-diazepane- 0.1 g of bis (ethane-2,1-diyl) bis (azanetriyl)) tetraacetic acid was dissolved in 2 ml of distilled water, and the mixture was dissolved in 10% thiophosgene / methylene chloride 1 ml was added thereto, followed by stirring for 2 hours. (S) -2,2 ', 2'',2''' - (((2- (4-fluorophenyl) Diazepane-1,4-diyl) bis (ethane-2,1-diyl)) bis (azanetriyl)) tetraacetic acid, which was obtained by LC- Mass. In the negative mode, the M-1 molecular weight of 564 was obtained.

< Experimental Example  1> Measurement of isotope label efficiency

The isotopic labeling efficiency using a compound of the present invention, a heterocyclic functional chelator, was measured using an isotope ( ¹⁷⁶ Lu (n, γ) ¹⁷⁷ Lu) produced by the Korea Atomic Energy Research Institute. The chelator agent of Example 1 and the NCS type chelator agent of Example 2 were used for the labeling efficiency test.

The experimental method for isotopic labeling is as follows.

The compounds of Example 1 and Example 2 were each prepared at a concentration of 10 ^-9 mol / ml and 10 ^-8 mol / ml in 50 mM acetate buffer conditions (pH = 5.5). Lu-177 (1 mCi) was added to each, and the mixture was stirred at room temperature for 1 minute. Thereafter, 1 μl of each solution was taken and the degree of labeling was confirmed using ITLC using 75% methanol as a developing solvent. The results are shown in FIG.

As shown in FIG. 1, when 75% methanol was used as the developing solvent, it was confirmed that the compound not labeled with rutethium (Lu) stays at the spotting point. In the saline condition, Stay at the front of the solvent front. In addition, as shown in FIGS. 2 and 3, the compound of Example 1 prepared at a concentration of 10 ^-9 mol / ml was labeled with a yield of 73%, and was produced at a concentration of 10 ^-8 mol / ml In the case of the compound of Example 2, the labeling yield was 97% or more.

Therefore, it can be confirmed that the labels of Examples 1 and 2 according to the present invention are efficiently labeled with ruthenium.

< Experimental Example  2> Biocompatible  Isotope labeling of antibodies

Step 1: Example  Of compound and antibody Biocompatibility

To label the antibody for isotopes, the compound of Example 2 according to the present invention was first conjugated to the antibody and the isotope was labeled.

The compound of Example 2 was stirred for 2 minutes under the condition of 50 mM PBS buffer to be introduced into the IgG antibody, and then cultured at room temperature for 24 hours. Labeling of lutein (Lu-177) was performed to confirm the progress of bioadhesion.

As shown in FIGS. 4A, 4B, and 4C, when the ratio of the compound of Example 2 and the antibody (IgG) was varied, it was confirmed that the amount of the labeled antibody was decreased as the amount of the compound was increased.

Step 2: Bonding  After doing Of lutein (Lu-177)  sign

2 mg of IgG as an antibody in the bioadhesive compound (Compound-IgG of Example 2) prepared in the above step 1 was used, and the bioadhesive compound used in the ratio of the compound of Example 2: antibody = 5: (Nap-10 column, GE Healthcare). 0.5 ml was taken from each of the vials 1 to 5 and separated on the SDS-page. As a result, it was confirmed that most of the bioadhesive compounds were present in the third vial as shown in FIG. 5.

A total of 0.7 ml of a 50 mM acetate buffer was added to the purified bioadhesive compound, and 0.5 mCi of rutemium 2 days after the production was added thereto, followed by stirring for 10 minutes. As a result of confirming the labeling efficiency of rutemium by ITLC as much as 3 상기 of the mixture, it was confirmed that the labeling efficiency was 98% or more as shown in FIG.

Therefore, it can be confirmed that the labeling of rutemium is efficiently performed after the bioadhesion with the antibody according to the present invention.

This is because the heterocyclic structure possessed by the compound according to the present invention plays a role of fixing the structure of the compound itself and consequently enhances the coordination effect with the metallic isotope ruthecium and the like. Therefore, the compounds according to the present invention can be effectively used for radioimmunotherapy by introducing them into various antibodies.

Claims (6)

Claims [1] A heterocyclic derivative represented by the following formula (1) or a pharmaceutically acceptable salt thereof:
[Chemical Formula 1]

(In the formula 1,
R is

or

ego;
X is an acetyl group;
and n is an integer of 1 to 2).
delete As shown in Scheme 1 below,
Reacting the compound of formula (2) as a starting material in a base and an organic solvent to prepare a compound of formula (3) (step 1);
Reacting the compound of Formula 3 prepared in Step 1 with NH ₃ in an organic solvent to prepare a compound of Formula 4 (Step 2);
Reacting the compound of Formula 4 prepared in Step 2 with an organic solvent to prepare a compound of Formula 5 (Step 3);
Reacting the compound of Formula 5 prepared in Step 3 with a compound of Formula 6 under a base to prepare a compound of Formula 7 (Step 4);
Reacting the compound of Formula 7 prepared in Step 4 with sodium salt in an organic solvent to prepare a compound of Formula 8 (Step 5);
Reacting the compound of Formula 8 prepared in Step 5 with a compound of Formula 9 in an organic solvent to prepare a compound of Formula 10 (Step 6);
Reacting the compound of formula 10 prepared in step 6 with an acid to produce a compound of formula 11 (step 7);
Reacting the compound of formula (11) prepared in step 7 with a compound of formula (12) in an organic solvent to prepare a compound of formula (13) (step 8);
Reacting the compound of formula 13 prepared in step 8 in an organic solvent to prepare a compound of formula 14 (step 9); And
A process for preparing a compound of formula (1) as defined in claim 1, comprising the step of reacting the compound of formula (14) prepared in step 9 with an acid and an organic solvent to prepare a compound of formula (1)
[Reaction Scheme 1]

(In the above Reaction Scheme 1,
R is

ego,
X and n are the same as defined in formula (1) of claim 1,
R ¹ is

ego,
R ² is a straight or branched alkyl of C ₁ ~ C _5).
A complex of a radioisotope metal capable of forming a coordination bond with the compound of claim 1 and a non-covalent electron pair of a nitrogen atom existing on the ring of the compound.
5. The complex of claim 4, wherein the metal is lutetium (Lu-177).
A medicament for radioimmunotherapy comprising the complex of claim 4 as an active ingredient.

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