KR101229929B1 - Novel maleimide compound having 1,2,3-triazole group, preparation method thereof and Ｆ­18 labeling method of biomolecule using prosthetic group - Google Patents

Abstract

The present invention relates to a radioactive isotope ¹⁸ F label of a maleimide compound labeled with fluorine-18 ( ¹⁸ F), a method for preparing the same, and a biological compound using the same as a supplementary group, wherein the compound of the present invention is labeled with ¹⁸ F. Easily prepared by the [3 + 2] ring reaction under a copper-catalyst between an azide compound and a maleimide acetylene compound, shortening the existing three or more complex manufacturing processes and reducing the production time. In addition, ¹⁸ F (half-life 110 minutes), a positron emitting radioisotope, may be useful for the effective synthesis of labeled subgroups.

Description

Novel maleimide compound having 1,2,3-triazole group, preparation method about and F18 labeling method of biomolecule using prosthetic group}

The present invention relates to a novel maleimide compound having a 1,2,3-triazole group, a method for preparing the same, and a method for labeling F-18 of a biological compound having the same as a supplement group.

Positron emission tomography (PET) is a 3-D imaging of the human body in real time using a molecular probe labeled with a radioisotope that emits positrons, providing information about the presence and progression of disease. As diagnostic molecular imaging technology applicable to human body, it is showing the biggest trend in diagnostic imaging field. Of the many positrons that emit positrons, fluorine-18 ( ¹⁸ F) is easily produced in high purity in medical particle accelerators and is very useful for positron emission tomography, given the overall manufacturing process / time including labeling reactions and radioactive exposure of patients. The radionuclide is the most studied because of its suitable properties. In general, the labeling of ¹⁸ F for low molecular organic synthetic compounds is through the leaving group of the sulfonate compound and a nucleophilic substitution reaction, which is carried out vigorously for 100 to 10-30 minutes in an aprotic polar solvent. The reaction nature of potassium carbonate (K ₂ CO _3), tetra - n - butylammonium bicarbonate _{(tetra- n -butylammonium bicarbonate, TBAHCO 3} ) or tetra - n - butylammonium hydroxide (tetra- n -butylammonium hydroxide, A large amount of base such as TBAOH) is used. Such basic vigorous labeling reactions can degrade even highly stable low molecular organic synthetic compounds, leading to degradation and epimerization of peptide-based proteins, nucleotides and the like, especially with weak chemical bonds. Therefore, labeling the ¹⁸ F biomaterial requires very mild reaction conditions. For this purpose, ¹⁸ F labeling compounds having reactive functional groups capable of binding to biological compounds under mild conditions are used (Okarvi, s. M. European) . Journal of Nuclear Medicine , 2001, 28, 929-938).

Maleimide functional groups are well known to cause a 1,4-Michael addition reaction with a compound having an SH-functional group, and are used as a method for binding a specific compound to a protein of a biological compound composed of peptides using the same. Shiue, CY groups are N- (4- [ ¹⁸ F] fluorophenyl) maleimide ([ ¹⁸ F] FPM) and N- [3- (2,5-di) labeled with ¹⁸ F as shown in the formula Oxo-2,5-dihydrophenol-1-yl) -phenyl] -4- [ ¹⁸ F] fluorobenzamide ([ ¹⁸ F] DDPFB) has been reported as a complement group that can label peptide biocompounds ( Shiue et al, Journal of Labeled Compounds and Radiopharmaceuticals , 26 , 287-289, 1989). However, this method requires a multi-step synthesis, which has a long manufacturing time and a low final radiochemical yield.

WO 2003/080544 discloses a method for synthesizing a compound having SH at its ¹⁸ F-labeled end, and then binding the peptide to the maleimide-treated peptide, thereby labeling the ¹⁸ F with the peptide. There is a disadvantage that a cumbersome pretreatment process for binding maleimide to the peptide compound is required.

In WO 2004/002984 a 2- [ ¹⁸ F] fluoropyridine-based maleimide subsidiary group ([ ¹⁸ F] FPyME) represented by the following formula has been developed and is still used today (de Bruin, B. et al, Bioconjugate). Chemistry , 16, 406-420, 2005), which also requires a long synthesis process of 3-4 steps, resulting in long manufacturing time and low radiochemical yield.

Tatsushi Toyokuni group is a new maleimide subsidiary group represented by N- {4-[(4- [ ¹⁸ F] fluorobenzylimidene) amino] butyl} maleimide ([ ¹⁸ F] FBABM) compound represented by the following formula Was developed and applied to glutathione (GSH) compounds with SH functionality (Toyokuni, T. et al, Bioconjugate Chemistry , 14, 1253-1259, 2003). The ^[18 F] FBABM compound is a ¹⁸ F-labeled 4- ^[18 F] fluoro-benzaldehyde ^[18 F] FBA) and N- [4-(aminoxy) butyl] Two through reaction with maleimide Synthesized in stages, has the advantage of having a short synthesis stage compared to the existing method, but the overall manufacturing time is a long and still has a disadvantage of showing a low radiochemical yield. As a further improved synthetic method, hexyl group-substituted supplemental group ([ ¹⁸ F] FBAHM) has been reported and has been applied to date (Berndt, M. et al, Nuclear Medicine and Biology , 34, 5-15). , 2007; Hultsch C., Applied Radiation and Isotopes , 65 , 818-826, 2007; Wuest, F., Bioconjugate Chemistry , 19, 1202-1210, 2008; Li, X., Bioconjugate Chemistry , 19, 1684-1688, 2008).

In addition, the Cai group introduced N- [2- (4- ¹⁸ F-fluorobenzaido) ethyl] maleimide ([ ¹⁸ F] FBEM) as a new complement group and added to cRGDyK compound substituted with a thiol group. (Cai, W., The Journal of Nuclear Medicine , 47, 1172-1180, 2006). The manufacturing method of the [ ¹⁸ F] FBEM compound has a disadvantage that the final radiochemical yield is only 5% ± 2% (nondecay-corrected) with a preparation time of 150 ± 20 minutes after three or more steps. [ ¹⁸ F] FBEM has also been applied to label ¹⁸ F in HER2-specific artificial binding proteins (Kiesewetter, DO, Journal of Fluorine Chemistry , 129, 799-806, 2008; Kramer-Marek, G., The Journal of Nuclear Medicine , 50, 1131-1139, 2009).

[3 + 2] ring reaction between azide and acetylene under copper (I) catalyst is the most representative reaction of Click Chemistry. It has very high reaction selectivity, proceeds rapidly at room temperature, and reacts well in aqueous solution. (Kolb, H., Angew. Chem. Int. Ed . 40, 2004-2021, 2001; Kolb HC, Drug Discovery Tody , 24, 1128-1137, 2003; Bock, VD, European Journal of Organic Chemistry , 51-68, 2006; Kurpiers, T., Angew. Chem. Int. Ed . 48, 1729-1731, 2009; Best.MD, Biochemistry , 48, 6571-6584, 2009). These reaction advantages have been used in many ways, and recently, ¹⁸ F has been studied as a method for labeling biological compounds (Marik, J., Tetrahedron). Letters , 47, 6681-6684, 2006; Sirion, U., Tetrahedron Letters , 48, 3953-3957, 2007; Glaser, M., Journal of Labeled Compounds and Radiopharmaceuticals , 52, 407-414, 2009).

Therefore, the present inventors prepared a novel maleimide supplementary group while researching to develop a compound capable of easily labeling ¹⁸ F on a biological compound, and the maleimide supplementary group has a cysteine amino acid residue on its surface. It was confirmed that the biological compound labeled with the radioisotope can be easily synthesized by combining the protein with Michael-addition reaction, and completed the present invention.

An object of the present invention is to provide a novel maleimide compound having a 1,2,3-triazole group, a method for preparing the same, and a method for labeling F-18 of a biological compound having the same as a prosthetic group.

In order to achieve the above object, the present invention provides a maleimide compound having a 1,2,3-triazole group represented by the following formula (1).

[Formula 1]

Wherein X, Y and F are as defined herein.

The present invention also provides a method for producing a maleimide compound having a 1,2,3-triazole group.

Furthermore, the present invention provides a method for producing a maleimide compound having a 1,2,3-triazol asleep labeled with radioisotope ¹⁸ F.

The present invention also provides a method of labeling a radioisotope ¹⁸ F in a biocompound.

The present invention provides a method for preparing ¹⁸ F-labeled maleimide subgroups in high radiochemical yield using alkyne / azide [3 + 2] ring reaction under a copper catalyst to shorten the synthesis step and production time. In addition, the quantitative compound synthesis through click chemistry can be enabled to significantly increase the radiochemical yield. In addition, the maleimide prosthetic group of the present invention can be used for labeling biochemical compounds, and in particular, a radioactive isotope-labeled biochemical compound can be bound to Michael-added reaction with a protein having a cysteine amino acid residue on its surface. Can be easily synthesized. Furthermore, the simplified prosthesis group manufacturing method of the present invention is well suited for the synthesis of high radiation doses by an automatic synthesis device.

1 is a schematic conceptual diagram of the present invention.
2 is a Radio-TLC diagram of a compound labeled with ¹⁸ F in step 1 of Example 4 according to the present invention.
3 is a Radio-TLC diagram of the compound labeled with ¹⁸ F in step 1 of Example 4 according to the present invention.
4 is a Radio-TLC diagram of a compound labeled with ¹⁸ F in step 2 of Example 4 according to the present invention.
FIG. 5 is a Radio-TLC diagram of the compound labeled ¹⁸ F in Example 5 according to the present invention. FIG.
6 is a Radio-TLC diagram of the compound labeled ¹⁸ F in Example 6 according to the present invention.
7 is a graph showing the HPLC analysis of the compound prepared in Example 6 according to the present invention.
8 is a Radio-TLC diagram of the compound labeled with ¹⁸ F of Experimental Example 1 according to the present invention.
9 is a Radio-TLC diagram of the compound labeled with ¹⁸ F of Experimental Example 2 according to the present invention.

Hereinafter, the present invention will be described in detail.

In order to achieve the above object, the present invention provides a maleimide compound having a 1,2,3-triazole group represented by the following formula (1).

In Formula 1,

X is a C ₁ -C ₆ straight or branched alkyl group; Or-[CH ₂ CH ₂ O] _n CH ₂ CH _2- ,

Y is a single bond, C ₁ -C ₆ straight or branched alkyl group; Or -CH ₂ [OCH ₂ CH ₂ ] _n- ,

n is an integer from 1 to 5,

F is ¹⁸ F or ¹⁹ F.

Preferably,

X is ethyl; n -propyl; Or-[CH ₂ CH ₂ O] _n CH ₂ CH _2- (n is an integer of 1-3),

Y is a single bond; methyl; ethyl; n -propyl; Or -CH ₂ [OCH ₂ CH ₂ ] _n- (n is an integer from 1-4),

F is ¹⁸ F or ¹⁹ F.

Preferred examples of the maleimide compound having a new 1,2,3-triazole group represented by Chemical Formula 1 according to the present invention are as follows.

(1) 1- {2- {2- [2- (fluoroethoxy) ethoxy] ethoxy} ethyl} -4- (N-maleimidylmethyl) -1,2,3-triazole;

(2) 1- {2- {2- [2- (2-fluoroethoxy) ethoxy] ethoxy} ethyl} -4- (3- (N-maleimidyl) propyl) -1,2, 3-triazole; And

(3) 1- {2- {2- [2- (2-fluoroethoxy) ethoxy] ethoxy} ethyl} -4-{{2- {2- [2- (N-maleimidyl) Ethoxy] ethoxy} ethoxy} methyl} -1,2,3-triazole.

The structural formulas of the compounds according to the present invention are shown in Table 1 below.

compound constitutional formula


One

2

3

The present invention also provides a method for preparing a maleimide compound having a 1,2,3-triazole group contained in the compound of Formula 1.

Specifically, the method for preparing a maleimide compound having a 1,2,3-triazole group is represented by Chemical Formula 3 by performing a nucleophilic fluorination reaction in an organic solvent with a sulfonate precursor represented by Chemical Formula 2, as shown in Scheme 1 below. Obtaining a fluoroalkyl azide compound as indicated (step 1); And

Reacting the fluoroalkyl azide compound of formula 3 obtained in step 1 with maleimide having a terminal alkyne group represented by formula 4 under an organic solvent and a copper catalyst to obtain a compound represented by formula 1 (step 2) do.

[Reaction Scheme 1]

(In Scheme 1, X and Y are as defined in Formula 1,

R is methyl, trifluoromethyl, p -toluenyl, p -nitrophenyl, soluble polyethylene glycol, polystyrene or insoluble polystyrene, preferably methyl, p -toluenyl or p -nitrophenyl)

In the method for producing a maleimide compound having a 1,2,3-triazole group of the present invention, Step 1 of Scheme 1 is a sulfonate precursor represented by the formula (2) is represented by the formula (3) through a nucleophilic fluorination reaction A step of synthesizing a fluoroalkyl azide compound. The organic solvent may be any one selected from the group consisting of acetonitrile, dimethylformamide (N, N-dimethylformamide, DMF), dimethylsulfoxide (dimethylsulfoxide, DMSO), tertiary alcohol, and a mixed solvent thereof. . Fluoride may be tetrabutylammonium fluoride (TBAF) or cesium fluoride (CsF).

Specifically, the sulfonate precursor represented by Chemical Formula 2 and cesium fluoride are added to a reaction vessel, and t -amyl alcohol is added, followed by stirring at 100 ° C. for about 3-6 hours, followed by column chromatography, which is represented by Chemical Formula 3 A fluoroalkyl azide compound can be obtained.

In the production method according to the invention, when F is ¹⁸ F, it can be labeled with ¹⁸ F using a polymer cartridge.

The ¹⁸ F labeling method of the present invention can be variously performed depending on whether a polymer cartridge is used.

First, in the case of using a polymer cartridge, an aqueous solution of [ ¹⁸ F] fluoride / [ ¹⁸ O] H ₂ O produced in cyclotron was converted into Chromafix ^? Or QMA ^? Pass the cartridge through the [ ¹⁸ F] fluoride to the cartridge and remove the water in the cartridge with methanol. A methanol solution containing TBAHCO ₃ or TBAOMs is run through the cartridge to elute the [ ¹⁸ F] fluoride trapped in the cartridge. Nitrogen gas is blown into this solution and the water and methanol solvent are completely removed for 1 to 3 minutes while heating to 100 ~ 120 ℃. The precursor compound for labeling with ¹⁸ F of Formula 2 prepared above is added to the reaction vessel and the reaction solvent is added to dissolve. The dissolved reaction mixture is heated at 100 to 130 ° C. for 3 to 30 minutes, cooled to room temperature, diluted with ethyl acetate, and passed through a silica SepPak cartridge. The silica SepPak cartridge was further washed with ethyl acetate, and the combined solution was removed by heating with nitrogen gas blowing at 50 -100 ° C. Solvent free residue is carried out in step 2 of Scheme 1 without separation. As the reaction solvent, acetonitrile, t -butanol, t -amyl alcohol, or a mixed solvent thereof may be used.

If no polymer cartridge is used, an aqueous solution of [ ¹⁸ F] fluoride / [ ¹⁸ O] H ₂ O produced from cyclotron is added to the reaction vessel, TBAHCO ₃ or TBAOH is added, and then acetonitrile to remove moisture Add nitrogen gas and heat to 100 ~ 120 ℃ by blowing nitrogen gas. The process of adding and removing acetonitrile is repeated three to four times until water is completely removed. The precursor compound for labeling with ¹⁸ F of Formula 2 prepared above is added to the reaction vessel and the reaction solvent is added to dissolve. The dissolved reaction mixture is heated at 100 to 130 ° C. for 3 to 30 minutes, cooled to room temperature, diluted with ethyl acetate, and passed through a silica SepPak cartridge. The silica SepPak cartridge was further washed with ethyl acetate, and the collected solution was removed by blowing nitrogen gas and heating at 50 to 100 ° C. Solvent free residue is carried out in step 2 of Scheme 1 without separation. As the reaction solvent, acetonitrile, t -butanol, t -amyl alcohol, or a mixed solvent thereof may be used.

Next, step 2 is a step of synthesizing the target compound represented by the formula (1) through a [3 + 2] ring reaction under a copper catalyst between the fluoroalkyl azide compound obtained in step 1 and the maleimide having a terminal alkyne group The organic solvent may be any one selected from the group consisting of acetonitrile, dimethylformamide (DMF), dimethyl sulfoxide (DMSO), water, tertiary alcohol, and a mixed solvent thereof. The copper catalyst may be a copper catalyst having a oxidation number of 1 consisting of copper iodide, copper bromide, and copper chloride, and includes copper sulfate (CuSO ₄ ), copper acetate (Cu (OAc) ₂ ), and copper nitrate (Cu (NO ₃ ) ₂ ) A copper catalyst having a oxidation number of 2 consisting of copper trifluoromethanesulfonate (Cu (OTf) ₂ ) and copper oxide (CuO) can be used. When the copper catalyst having the oxidation number of 2 is used, any one reducing agent selected from the group consisting of sodium ascorbate, sodium sulfite (Na ₂ SO ₃ ), and dithiothreitol may be further used. Preferably, the copper catalyst may use CuI or CuSO ₄ / sodium ascorbate. The base may be added together when using a copper catalyst having an oxidation number of 1 and selected from the group consisting of metal salts of bicarbonate ions, alkali metal salts of carbonate ions, triethylamine, diisopropylethylamine, pyridine, lutidine and collidine Any one of them can be used, and preferably triethylamine or diisopropylethylamine can be used.

Specifically, the maleimide compound having the intermediate of Formula 3 produced in Step 1 and the terminal alkyne group represented by Formula 4 was added to a reaction vessel, dissolved by adding acetonitrile, and copper iodide and triethylamine were added. After stirring until complete, column chromatography may be performed to obtain the target compound of Chemical Formula 1.

In the manufacturing method according to the present invention, when F is ¹⁸ F in Step 2, the reaction is carried out by adding a maleimide compound having a terminal alkyne group of Formula 4 to the intermediate of Formula 3 labeled with ¹⁸ F produced in Step 1 After dissolving by adding a solvent, copper iodide and triethylamine were added, followed by stirring at room temperature for 5-20 minutes, and the reaction mixture was passed through an imide compound pPak cartridge to remove the copper catalyst, followed by high efficiency column chromatography (HPLC). The target compound of 1 can be isolate | separated.

In addition, the present invention, by using the ^[18 F] compounds of the general formula (1) labeled with ¹⁸ F, to provide a method for labeling a ^bio-18 F in the compound with a SH- functional groups, such as Scheme 2.

Scheme 2

(X and Y of the reaction scheme is as defined in Formula 1)

Compound 6 of Scheme 2 may be obtained by adding the compound labeled with ¹⁸ F obtained in Scheme 1, dissolved in PBS buffer solution in an organic solvent, and stirred.

The preparation of a maleimide compound ¹⁸ F on the cover according to the present invention can be prepared in high radiochemical yield, shorten the synthetic steps, and manufacturing time, the produced maleimide compound is a cysteine (cystein) amino acid residue on the surface Because of the Michael-addition reaction with the protein shown, the radioisotope-labeled biological compound can be easily synthesized.

Hereinafter, the present invention will be described in detail by production examples, examples and experimental examples. However, the content of the present invention is not limited by the following Production Examples, Examples and Experimental Examples.

< Manufacturing example  1> Diethylene glycol  Mono- Methanesulfonate  Mono- Of azide (2a)  Produce

Diethylene glycol dimethanesulfonate ( 7 , 3.00 g, 11.44 mmol) and sodium azide (817 mg, 12.58 mmol) were added to the reaction vessel, acetonitrile (50 mL) was added, and the reaction mixture was stirred at 80 DEG C for 24 hours. Stirred. The precipitate was filtered off and then the filtrate was subjected to column chromatography (40% ethyl acetate / n -hexane) to give the title compound 2a (838 mg, 35%) as a colorless liquid.

¹ H NMR (CDCl ₃ , 200 MHz) δ 3.07 (s, 3H), 3.41 (t, J = 4.8 Hz, 2H), 3.70 (t, J = 4.8 Hz, 2H), 3.76-3.81 (m, 2H) , 4.35-4.40 (m, 2 H); ¹³ C NMR (CDCl ₃ , 50 MHz) δ 37.3, 50.5, 68.7, 68.8, 69.9.

< Manufacturing example  2> Triethylene  Glycol mono- Toluenesulfonate 10 and d- Of toluenesulfonate (12)  Produce

Triethylene glycol ( 8 , 13.3 mL, 99.9 mmol) and para-toluenesulfonyl chloride (19.04 g, 99.9 mol) were dissolved in pyridine (150 mL), and the reaction mixture was stirred at 0 ° C. for 7 hours, followed by water And 4 M sulfuric acid were added and the organic compound was extracted with ethyl acetate. The extracted ethyl acetate solution was treated with anhydrous sodium sulfate, and then subjected to column chromatography (80% ethyl acetate / n -hexane) to obtain the target compounds 10 (14.8 g, 47%) and 12 (8.15 g, 17%). .

Compound 10: ¹ H NMR (200 MHz, CDCl ₃ ) δ 2.45 (s, 3H), 3.55-3.61 (m, 6H), 3.68-3.73 (m, 4H), 4.15-4.20 (m, 2H), 7.35 ( d, J = 8.6 Hz, 2H), 7.80 (d, J = 8.4 Hz, 2H); ¹³ C NMR (50 MHz, CDCl ₃ ) δ 21.6, 61.7, 68.6, 69.1, 70.2, 70.7, 72.4, 127.9 (2C), 129.8 (2C), 132.9, 144.8.

Compound 12: ¹ H NMR (200 MHz, CDCl ₃ ) δ 2.45 (s, 6H), 3.53 (s, 4H), 3.66 (t, J = 4.8 Hz, 4H), 4.14 (t, J = 4.8 Hz, 4H ), 7.34 (d, J = 8.6 Hz, 4H), 7.79 (d, J = 8.4 Hz, 4H); ¹³ C NMR (50 MHz, CDCl ₃ ) δ 21.6, 68.7, 69.2, 70.7, 127.9, 129.8, 133.0, 144.8.

< Manufacturing example  3> Tetraethylene  Glycol mono- Toluenesulfonate 11 and Of dietoluenesulfonate (13)  Produce

Tetraethylene glycol ( 9 , 18.0 mL, 0.104 mol) and para-toluenesulfonyl chloride (19.9 g, 0.105 mol) were dissolved in pyridine (200 mL), and then the reaction mixture was stirred at 0 ° C. for 7 hours, followed by water And 4 M sulfuric acid were added and the organic compound was extracted with ethyl acetate. The extracted ethyl acetate solution was treated with anhydrous sodium sulfate, and then subjected to column chromatography (80% ethyl acetate / n -hexane) to obtain the target compounds 11 (16 g, 45%) and 13 (31 g, 20%). .

Compound 11: ¹ H NMR (400 MHz, CDCl ₃ ) δ 2.45 (s, 3H), 3.59-3.73 (m, 14H), 4.17 (t, J = 4.8 Hz, 2H), 7.35 (d, J = 8.4 Hz , 2H), 7.80 (d, J = 8.4 Hz, 2H); ¹³ C NMR (100 MHz, CDCl ₃ ) δ 21.9, 61.9, 68.9, 69.5, 70.5, 70.6, 70.8, 70.9, 72.7, 128.2, 130.1, 133.1, 145.1.

Compound 13: ¹ H NMR (400 MHz, CDCl ₃ ) δ 2.45 (s, 6H), 3.57 (s, 8H), 3.68 (t, J = 4.8 Hz, 4H), 4.15 (t, J = 4.8 Hz, 4H ), 7.34 (d, J = 6.0 Hz, 4H), 7.79 (d, J = 6.0 Hz, 4H); ¹³ C NMR (100 MHz, CDCl ₃ ) δ 21.9, 68.9, 69.5, 70.8, 71.0, 128.2, 130.1, 133.2, 145.0.

< Manufacturing example  4> Triethylene  Glycol mono- Azide (14)  Produce

Compound 10 (14.4 g, 45.1 mmol) obtained in Preparation Example 2 was dissolved in acetonitrile (140 mL), and sodium azide (4.40 g, 67.7 mmol) and tetrabutylammonium iodide (3.33 g, 9.03 mmol) were added thereto. It was. 60 reaction mixtures After stirring for 20 hours at ℃, the filtered precipitate was washed with acetonitrile. Water and saturated aqueous sodium chloride solution were added to the filtrate, and the organic compound was extracted with ethyl acetate. The extracted ethyl acetate solution was treated with anhydrous sodium sulfate and then column chromatography (60% ethyl acetate / n -hexane) was carried out to obtain the desired compound 14 (5.60 g, 84%).

¹ H NMR (200 MHz, CDCl ₃ ) δ 2.39 (br s, 1H), 3.41 (t, J = 4.9 Hz, 2H), 3.60-3.74 (m, 10H); ¹³ C NMR (50 MHz, CDCl ₃ ) δ 50.6, 61.7, 70.0, 70.4, 70.6, 72.5.

< Manufacturing example  5> Tetraethylene  Glycol mono- Azide (15)  Produce

Compound 11 (3.00 g, 8.61 mmol) obtained in Preparation Example 3 was dissolved in acetonitrile (60 mL), followed by sodium azide (1.05 mg, 17.2 mmol) and tetrabutylammonium iodide (TBAI, 636 mg, 1.72 mmol). Was added. The reaction mixture was stirred at 60 ° C. for 20 hours, filtered and the filtered precipitate was washed with acetonitrile. Water and saturated aqueous sodium chloride solution were added to the filtrate, and the organic compound was extracted with ethyl acetate. The extracted ethyl acetate solution was treated with anhydrous sodium sulfate and then column chromatography (80% ethyl acetate / n -hexane) was carried out to obtain the desired compound 15 (1.70 g, 92%).

¹ H NMR (400 MHz, CDCl ₃ ) δ 2.70 (br s, 1H), 3.36 (t, J = 4.8 Hz, 2H), 3.57 (t, J = 4.6 Hz, 2H), 3.61-3.66 (m, 10H ), 3.69 (t, J = 4.4 Hz, 2H); ¹³ C NMR (100 MHz, CDCl ₃ ) δ 50.9, 61.9, 70.24, 70.52, 70.77, 70.85, 70.89, 72.7.

< Manufacturing example  6> Triethylene  Glycol mono- Toluenesulfonate  Mono- Of azide (2b)  Produce

Compound 14 (7.97 g 54.2 mmol) and para-toluenesulfonyl chloride (10.3 g, 54.2 mmol) obtained in Preparation Example 4 were dissolved in pyridine (80 mL), and the reaction mixture was stirred at 0 ° C. for 9 hours. After adding water and 4 M sulfuric acid, the organic compound was extracted with ethyl acetate. The extracted ethyl acetate solution was treated with anhydrous sodium sulfate and then subjected to column chromatography (80% ethyl acetate / n -hexane) to give the title compound 2b (14.50 g, 89%).

¹ H NMR (200 MHz, CDCl ₃ ) δ 2.45 (s, 3H), 3.3 (t, J = 5.1 Hz, 2H), 3.61 (s, 4H), 3.62-3.73 (m, 4H), 4.1 (t, J = 4.7 Hz, 2H), 7.35 (d, J = 8.0 Hz, 2H), 7.80 (d, J = 8.4 Hz, 2H); ¹³ C NMR (100 MHz, CDCl ₃ ) δ 21.6, 50.6, 68.7, 69.2, 70.0, 70.6, 70.7, 127.9, 129.8, 133.0, 144.8.

< Manufacturing example  7> Tetraethylene  Glycol mono- Toluenesulfonate  Mono- Of azide (2c)  Produce

Compound 15 (1.50 g 6.84 mmol) and para-toluenesulfonyl chloride (1.3 g, 6.84 mmol) obtained in Preparation Example 5 were dissolved in pyridine (150 mL), and the reaction mixture was stirred at 0 ° C. for 9 hours. , Water and 4 M sulfuric acid were added, and then the organic compound was extracted with ethyl acetate. The extracted ethyl acetate solution was treated with anhydrous sodium sulfate and then subjected to column chromatography (80% ethyl acetate / n -hexane) to obtain the title compound 2c (1.60 g, 63%).

¹ H NMR (400 MHz, CDCl ₃ ) δ 2.43 (s, 3H), 3.36 (t, J = 4.8 Hz, 2H), 3.58 (s, 4H), 3.62-3.68 (m, 8H), 4.14 (t, J = 4.4 Hz, 2H), 7.32 (d, J = 8.4 Hz, 2H), 7.78 (d, J = 8.0 Hz, 2H); ¹³ C NMR (100 MHz, CDCl ₃ ) δ 21.9, 50.9, 68.9, 69.5, 70.3, 70.8, 70.9, 70.9, 71.0, 128.2, 130.0, 133.2, 145.0.

< Manufacturing example  8> Triethylene  Glycol mono- Toluenesulfonate  Mono- Of azide (2b)  Produce

Compound 12 (8.04 g, 17.02 mmol) obtained in Preparation Example 2 was dissolved in acetonitrile (80 mL), and sodium azide (1.11 g, 17.02 mmol) and tetrabutylammonium iodide (1.26 g, 3.40 mmol) were added thereto. It was. 60 reaction mixtures After stirring for 20 h at rt, the reaction mixture was filtered and washed with acetonitrile. Water and saturated aqueous sodium chloride solution were added to the filtrate, and the organic compound was extracted with ethyl acetate. The extracted ethyl acetate solution was treated with anhydrous sodium sulfate and then subjected to column chromatography (30% ethyl acetate / n -hexane) to give the title compound 2b (570 mg, 11%).

¹ H NMR and ¹³ C NMR are the same as in Preparation Example 6.

< Manufacturing example  9> Tetraethylene  Glycol mono- Toluenesulfonate  Mono- Of azide (2c)  Produce

Compound 13 (3.58 g, 7.12 mmol) obtained in Preparation Example 3 was dissolved in acetonitrile (55 mL), and sodium azide (400 mg, 6.55 mmol) and tetrabutylammonium iodide (483.9 mg, 1.31 mmol) were added thereto. It was. After the reaction mixture was stirred at 60 ° C. for 20 hours, the reaction mixture was filtered and the filtered precipitate was washed with acetonitrile. Water and saturated aqueous sodium chloride solution were added to the filtrate, and the organic compound was extracted with ethyl acetate. The extracted ethyl acetate solution was treated with anhydrous sodium sulfate and then subjected to column chromatography (60% ethyl acetate / n -hexane) to give the title compound 2c (479 mg, 18%).

¹ H NMR and ¹³ C NMR are the same as in Preparation Example 7.

< Manufacturing example  10> 4- Pentin -One- Of toluenesulfonate (16)  Produce

4-pentin-1-ol (0.7 mL, 7.56 mmol) and para-toluenesulfonyl chloride (1.44 g, 7.56 mmol) were dissolved in pyridine (13 mL), and the reaction mixture was stirred at room temperature for 3 hours. Water and 4 M sulfuric acid were added, and the organic compound was extracted with ethyl acetate. The extracted ethyl acetate solution was treated with anhydrous sodium sulfate, and then subjected to column chromatography (20% ethyl acetate / n -hexane) to obtain the target compound 16 (1.30 g, 72%).

¹ H NMR (500 MHz, CDCl ₃ ) δ 1.89-1.84 (m, 3H), 2.26 (dt, J = 7.0, 3.0 Hz, 2H), 2.45 (s, 3H), 4.15 (t, J = 6.0 Hz, 2H), 7.35 (d, J = 8.5 Hz, 2H), 7.80 (d, J = 9.0 Hz, 2H); ¹³ C NMR (125 MHz, CDCl ₃ ) δ 14.7, 21.6, 27.7, 68.9, 69.4, 82.1, 127.9, 129.8, 133.0, 144.8.

< Manufacturing example  11> Triethylene  Glycol mono- Of propazyl (17)  Produce

Sodium hydride (266 mg, 6.66 mol) was dissolved in anhydrous tetrahydrofuran (40 mL), and triethylene glycol 8 (1.78 mL, 13.3 mmol) was slowly added at -10 ° C. After stirring well at room temperature for 30 minutes, propazyl chloride (0.482 mL, 6.66 mmol) was added slowly. After stirring for 3 hours at room temperature, water was slowly added to terminate the reaction, and the organic compound was extracted with ethyl acetate. The extracted ethyl acetate solution was washed with saturated aqueous sodium chloride solution, treated with anhydrous sodium sulfate, and then column chromatography ( 100% ethyl acetate) was performed to obtain target compound 17 (1.80 g, 72%).

¹ H NMR (300 MHz, CDCl ₃ ) δ 2.44 (t, J = 2.1 Hz, 1H), 3.60-3.63 (m, 3H), 3.66-3.75 (m, 10H), 4.21 (d, J = 2.4 Hz, 2H); ¹³ C NMR (125 MHz, CDCl ₃ ) δ 58.5, 61.8, 63.7, 68.9, 70.4, 70.7, 72.6, 74.8, 79.7.

< Manufacturing example  12> Triethylene  Glycol mono- Propazyl  Mono- Of methanesulfate (18)  Produce

Compound 17 (22.0 g, 0.117 mol) and diisopropylethylamine (18.13 g, 0.14 mol) obtained in Preparation Example 11 were dissolved in dichloromethane (70 mL), and methanesulfonyl chloride (9.95 mL, 0.13 mol) was added slowly. Thereafter, the reaction mixture was raised to room temperature, stirred for 1 hour, water was added, and then the organic compound was extracted with ethyl acetate. The extracted ethyl acetate solution was treated with anhydrous sodium sulfate, and then subjected to column chromatography (80% ethyl acetate / n -hexane) to obtain target compound 18 (25.8 g, 83%).

¹ H NMR (500 MHz, CDCl ₃ ) δ 2.44 (t, J = 2.5 Hz, 1H), 3.09 (s, 3H), 3.65-3.70 (m, 8H), 3.77 (t, J = 4.5 Hz, 2H) , 4.20 (d, J = 2.5 Hz, 2H), 4.39 (t, J = 4.5 Hz, 2H); ¹³ C NMR (125 MHz, CDCl ₃ ) δ 37.9, 58.6, 69.2, 69.3, 69.5, 70.6, 70.7, 70.8, 74.8, 79.7.

< Manufacturing example  13> Bycycle Maleimide (19)  Produce

Furan (22.48 mL, 0.309 mmol) and maleimide (10.0 g, 0.103 mol) were added to diethyl ether (100 mL) and heated to reflux overnight. After cooling the reaction mixture to room temperature, the resulting precipitate was filtered, washed several times with n -hexane, and then decompressed to obtain the target compound 19 (15.0 g, 88%).

¹ H NMR (500 MHz, CDCl ₃ ) δ 2.89 (s, 2H), 3.56-3.57 (m, 2H), 5.31 (s, 2H), 5.33-5.34 (m, 2H), 6.51 (s, 2H), 6.52 (s, 2 H), 8.04 (br s, 1 H), 8.45 (br s, 1 H); ¹³ C NMR (125 MHz, CDCl ₃ ) δ 47.6, 48.9, 79.6, 81.1, 134.8, 136.8, 175.1, 176.4.

< Manufacturing example  14> N- Propazyl Bycycle Of maleimide (20a)  Produce

Dimethylformamide (4 mL) was added to compound 19 (300 mg, 1.82 mmol) and potassium carbonate (376.7 mg, 2.73 mmol) obtained in Preparation Example 13, and propazyl chloride (0.130 mL, 1.82 mmol) was added thereto. After the reaction mixture was stirred at 60 ° C. for 3 hours, water and ammonium chloride were added, and the organic compound was extracted with ethyl acetate. The extracted ethyl acetate solution was treated with anhydrous sodium sulfate, and then subjected to column chromatography (50% ethyl acetate / n -hexane) to obtain the target compound 20a (199 mg, 54%).

¹ H NMR (400 MHz, CDCl ₃ ) δ 2.21 (t, J = 2.6 Hz, 1H), 2.91 (s, 2H), 4.24 (d, J = 2.4 Hz, 2H), 5.31 (d, J = 0.8 Hz , 2H), 6.53 (t, J = 0.8 Hz, 2H); ¹³ C NMR (100 MHz, CDCl ₃ ) δ 27.9, 47.6, 71.5, 76.4, 80.9, 136.6, 174.8.

< Manufacturing example  15> N- (4- Pentin ) Bycycle Of maleimide (20b)  Produce

Preparative Compound 19 obtained in Example 13 (291 mg, 1.76 mmol) and potassium carbonate (365.3 mg, 2.64 mmol) dimethylformamide (4 mL) a put the compound obtained in Preparation Example 10 16 (420 mg, 1.76 mmol in ) Was added. After the reaction mixture was stirred at 60 ° C. for 3 hours, water and ammonium chloride were added, and the organic compound was extracted with ethyl acetate. The extracted ethyl acetate solution was treated with anhydrous sodium sulfate, and then subjected to column chromatography (50% ethyl acetate / n -hexane) to obtain the target compound 20b (261 mg, 64%).

¹ H NMR (400 MHz, CDCl ₃ ) δ 1.81 (quintet, J = 7.0 Hz, 2H), 1.98 (t, J = 2.8 Hz, 1H), 2.20 (dt, J = 7.2, 2.8 Hz, 2H), 2.84 (s, 2H), 3.59 (t, J = 7.2 Hz, 2H), 5.27 (d, J = 0.4 Hz, 2H), 6.49 (d, J = 0.4 Hz, 2H); ¹³ C NMR (100 MHz, CDCl ₃ ) δ 16.0, 26.3, 38.0, 47.4, 69.0, 80.9, 82.9, 136.5, 176.2.

< Manufacturing example  16> N- {2- [2- (2- Propazyloxy ) Ethoxy ] Ethoxy }ethyl Bycycle Maleimide Preparation of 20c

Sodium hydride (394 mg, 9.86 mmol) was dissolved in dimethylformamide (15 mL), and compound 19 (1.09 g, 6.57 mmol) obtained in Preparation Example 13, which was dissolved in dimethylformamide, was slowly added thereto, followed by dimethylformamide. Potassium iodide (1.09 g, 6.57 mmol) dissolved in and Compound 18 (1.75 g, 6.57 mmol) obtained in Preparation 12 were added. After the reaction mixture was stirred at room temperature for 7 days, water was added slowly, ammonium chloride was added, the organic compound was extracted with ethyl acetate, and the organic layer was washed with a saturated aqueous sodium chloride solution. The extracted ethyl acetate solution was treated with anhydrous sodium sulfate and then subjected to column chromatography (70% ethyl acetate / n -hexane) to obtain the target compound 20c (485 mg, 22%).

¹ H NMR (500 MHz, CDCl ₃ ) δ 2.43 (t, J = 2.3 Hz, 1H), 2.86 (s, 2H), 3.61-3.66 (m, 8H), 3.68-3.69 (m, 4H), 4.20 ( d, J = 2.5 Hz, 2H), 5.26 (s, 2H), 6.51 (s, 2H); ¹³ C NMR (125 MHz, CDCl ₃ ) δ 30.4, 47.7, 58.6, 67.3, 69.3, 70.3, 70.6, 70.8, 74.7, 79.9, 81.1, 136.7, 176, 3.

< Manufacturing example  17> N- Propazyl Of maleimide (4a)  Produce

Compound 20a (4.00 g, 19.68 mmol) obtained in Preparation Example 14 was dissolved in anisole (30 mL), the reaction mixture was heated to reflux overnight, and then heated at 150 ° C. after removal of the reflux apparatus to remove most of anisole. It was. Column chromatography (30% ethyl acetate / n -hexane) was carried out to give the desired compound 4a (1.73 g, 65%).

¹ H NMR (400 MHz, CDCl ₃ ) δ 2.22 (t, J = 2.4 Hz, 1H), 4.30 (d, J = 2.0 Hz, 2H), 6.77 (s, 2H); ¹³ C NMR (100 MHz, CDCl ₃ ) δ 26.8, 71.5, 76.9, 134.5, 169.3.

< Manufacturing example  18> N- (4- Pentin ) Of maleimide (4b)  Produce

The compound 20b (330 mg, 1.43 mmol) obtained in Preparation Example 15 was dissolved in anisole (1 mL), and the reaction mixture was heated to reflux overnight, and then heated to 150 ° C. to remove most of anisole from the reflux apparatus. It was. Column chromatography (30% ethyl acetate / n -hexane) was carried out to give the desired compound 4b (179 mg, 77%).

¹ H NMR (400 MHz, CDCl ₃ ) δ 1.84 (quintet, J = 7.2 Hz, 2H), 1.97 (t, J = 2.6 Hz, 1H), 2.23 (dt, J = 7.2, 2.4 Hz, 2H), 3.64 (t, J = 7.0 Hz, 2H), 6.71 (s, 2H); ¹³ C NMR (100 MHz, CDCl ₃ ) δ 16.1, 27.2, 37.0, 69.0, 82.9, 134.1, 170.7.

< Manufacturing example  19> N- {2- [2- (2- Propazyloxy ) Ethoxy ] Ethoxy }ethyl Maleimide Preparation of (4c)

20c (462 mg, 1.38 mmol) of the compound obtained in Preparation 16 was dissolved in anisole (1 mL), and the reaction mixture was heated to reflux overnight. After removing the reflux apparatus, heating was performed at 150 ° C. to remove most of anisole. It was. Column chromatography (70% ethyl acetate / n -hexane) was carried out to give the desired compound 4c (346 mg, 94%).

¹ H NMR (500 MHz, CDCl ₃ ) δ 2.43 (t, J = 2.5 Hz, 1H), 2.86 (s, 2H), 3.66-3.60 (m, 8H), 3.67-3.69 (m, 2H), 3.73 ( t, J = 5.8 Hz, 2H), 4.20 (d, J = 2.5 Hz, 2H), 6.70 (s, 2H); ¹³ C NMR (125 MHz, CDCl ₃ ) δ 37.1, 58.4, 67.8, 69.1, 70.0, 70.4, 70.5, 74.4, 79.7, 134.1, 170,8.

< Example  1>

Step 1-A: Triethylene  Glycol mono- Fluoro  Mono- Of azide (3b)  Produce

To a reaction vessel containing Compound 2b (3.00 g 10.0 mmol) and cesium fluoride (3.0 g, 20.0 mmol) obtained in Preparation Example 6 or 8 was added t -amyl alcohol (50 mL), and then the reaction mixture was 110 ° C. After stirring for 3 hours, water and saturated aqueous sodium chloride solution were added, and the organic compound was extracted with ethyl acetate. The extracted ethyl acetate solution was treated with anhydrous sodium sulfate and then subjected to column chromatography (15% ethyl acetate / n -hexane) to give the title compound 3b (1.10 g, 74%).

¹ H NMR (400 MHz, CDCl ₃ ) δ 3.40 (t, J = 4.9 Hz, 2H), 3.67-3.72 (m, 7H), 3.84 (t, J = 4.0, 1H), 4.58 (dt, J = 47.8 , 4.2 Hz, 2H); ¹³ C NMR (100 MHz, CDCl ₃ ) δ 50.6, 70.0, 70.4 (d, J = 19.7 Hz), 70.7, 70.8, 83.1 (d, J = 168.3 Hz).

Step 1-B: Tetraethylene  Glycol mono- Fluoro  Mono- Of azide (3c)  Produce

T -amyl alcohol (50 mL) was added to a reaction vessel containing Compound 2c (2.40 g 6.34 mmol) and cesium fluoride (1.75 g, 11.5 mmol) obtained in Preparation Example 7 or 9, and the reaction mixture was then heated to 110 ° C. After stirring for 3 hours, water and saturated aqueous sodium chloride solution were added and the organic compound was extracted with ethyl acetate. The extracted ethyl acetate solution was treated with anhydrous sodium sulfate and then subjected to column chromatography (50% ethyl acetate / n -hexane) to give the title compound 3c (1.11 g, 78%).

¹ H NMR (400 MHz, CDCl ₃ ) δ 3.37 (t, J = 5.2 Hz, 2H), 3.65-3.67 (m, 10H), 3.70 (t, J = 4.0 Hz, 1H), 3.77 (t, J = 4.0 Hz, 1H), 4.55 (dt, J = 48.0, 4.0 Hz, 2H); ¹³ C NMR (100 MHz, CDCl ₃ ) δ 50.9, 70.3, 70.6 (d, J = 20.1 Hz), 70.9, 71.0, 83.4 (d, J = 168.2 Hz).

Step 2: 1- {2- {2- [2- (2- Fluoroethoxy ) Ethoxy ] Ethoxy } Ethyl} -4- (N- Maleimidylmethyl ) -1,2,3-triazole ( 1a)  Produce

Compound 3c (57 mg, 0.26 mmol) obtained in step 1-B and compound 4a (35 mg, 0.26 mmol) obtained in Preparation Example 17 were dissolved in acetonitrile (0.8 mL), and then CuI (10 mg, 0.05 mmol) was dissolved. ) And triethylamine (54 μl, 0.39 mmol) were added. After the reaction mixture was stirred at room temperature for 1 hour, water was added and the organic compound was extracted with ethyl acetate. The extracted ethyl acetate solution was treated with anhydrous sodium sulfate and then subjected to column chromatography (5% methanol / dichloromethane) to obtain the title compound 1a (45 mg, 49%).

¹ H NMR (500 MHz, CDCl ₃ ) δ 3.60-3.66 (m, 6H), 3.68-3.73 (m, 3H), 3.78 (t, J = 3.4 Hz, 1H), 3.85 (t, J = 4.0 Hz, 2H), 4.50-4.53 (m, 3H), 4.61 (t, J = 3.2 Hz, 1H), 4.82 (s, 2H), 6.73 (s, 2H), 7.72 (s, 1H); ¹³ C NMR (125 MHz, CDCl ₃ ) δ 32.8, 50.2, 69.3, 70.3 (d, J = 19.8 Hz), 70.5, 70.7, 83.1 (d, J = 168 Hz), 123.7, 134.2, 142.4, 170.0.

< Example  2> 1- {2- {2- [2- (2- Fluoroethoxy ) Ethoxy ] Ethoxy } Ethyl} -4- (3- (N- Maleimidyl ) Propyl) -1,2,3- Of triazole (1b)  Produce

Compound 3c (203 mg, 0.92 mmol) obtained in step 1-B of Example 1 and compound 4b (150 mg, 0.92 mmol) obtained in Preparation Example 18 were dissolved in acetonitrile (2 mL), and then CuI (35) was used. mg, 0.18 mmol) and triethylamine (0.192 mL, 1.38 mmol) were added. After the reaction mixture was stirred at room temperature for 1 hour, water was added and the organic compound was extracted with ethyl acetate. The extracted ethyl acetate solution was treated with anhydrous sodium sulfate and then subjected to column chromatography (5% methanol / dichloromethane) to obtain the title compound 1b (177 mg, 50%).

¹ H NMR (500 MHz, CDCl ₃ ) δ 1.96 (quintet, J = 7.3 Hz, 2H), 2.69 (t, J = 7.5 Hz, 2H), 3.53-3.57 (m, 2H), 3.58-3.62 (m, 6H), 3.63-3.65 (m, 2H), 3.68 (t, J = 4.0 Hz, 1H), 3.84 (t, J = 5.5 Hz, 2H), 4.47-4.50 (m, 3H), 4.58 (t, J = 8.5 Hz, 1H), 6.68 (s, 2H), 7.54 (s, 1H); ¹³ C NMR (125 MHz, CDCl ₃ ) δ 23.0, 28.2, 37.2, 50.2, 69.6, 70.4 (d, J = 19.6 Hz), 70.6, 70.8, 83.2 (d, J = 168 Hz), 122.3, 134.1, 147.6 , 170.8.

< Example  3> 1- {2- {2- [2- (2- Fluoroethoxy ) Ethoxy ] Ethoxy } Ethyl} -4-{{2- {2- [2- (N- Maleimidyl ) Ethoxy ] Ethoxy } Ethoxy } methyl } -1,2,3- Of triazole (1c)  Produce

Compound 3c (41 mg, 0.19 mmol) obtained in step 1-B of Example 1 and compound 4c (50 mg, 0.19 mmol) obtained in Preparation Example 19 were dissolved in acetonitrile (1 mL), and then CuI (7 mg, 0.037 mmol) and triethylamine (39 μl, 0.28 mmol) were added. After the reaction mixture was stirred at room temperature for 1 hour, water was added and the organic compound was extracted with ethyl acetate. The extracted ethyl acetate solution was treated with anhydrous sodium sulfate and then column chromatography (5% methanol / dichloromethane) was carried out to obtain the title compound 1c (45 mg, 54%).

¹ H NMR (500 MHz, CDCl ₃ ) δ 3.59-3.62 (m, 3H), 3.62-3.65 (m, 11H), 3.67-3.69 (m, 4H), 3.71-3.74 (m, 3H), 3.77 (t , J = 4.5 Hz, 1H), 3.89 (t, J = 5.5 Hz, 2H), 4.52 (t, J = 4.0 Hz, 1H), 4.55 (t, J = 5.0 Hz, 1H), 4.61 (t, J = 4.0 Hz, 1H), 4.68 (s, 2H), 6.70 (s, 2H), 7.76 (s, 1H); ¹³ C NMR (125 MHz, CDCl ₃ ) δ 37.1, 50.2, 64.6, 67.8, 69.4, 69.6, 70.0, 70.4 (d, J = 19.6 Hz), 70.5, 70.6, 70.8, 83.1 (d, J = 168 Hz) , 123.8, 134.1, 145.0, 170.6.

< Example  4>

Step 1: Tetraethylene  Glycol mono- [ ¹⁸ F] Fluoro  Mono-azide ([ ¹⁸ F] 3c) Preparation

Chromafix ^? [ ¹⁸ F] fluoride (12.47 mCi) was placed on a PS-HCO ₃ ) cartridge and eluted with a mixed solution of 0.2 M TBAOMs methanol solution (0.5 mL) and 0.2 MK ₂ CO ₃ aqueous solution (0.2 mL) [ ¹⁸ F] Fluoride was eluted into the reaction vessel. The amount of [ ¹⁸ F] fluoride remaining in the cartridge was 0.33 mCi. The eluted solution was heated to 120 with blowing nitrogen to remove the solvent, and acetonitrile (0.5 mL) was added thereto and repeated until water was completely removed. Compound 2c (3 mg) and t -amyl alcohol (0.5 mL) obtained in Step 1-B of Example 1 were added to the reaction vessel, and the reaction mixture was stirred at 120 ° C. for 10 minutes and then cooled to room temperature. ¹⁸ F label yield using Radio-TLC was 80%. Ethyl acetate (1 mL) was added to the reaction vessel, diluted and passed through silica SepPak. The reaction vessel and silica SepPak were washed once and twice with ethyl acetate (0.5 mL), respectively. The solvent was removed by injecting nitrogen while heating the eluted solution to 120 ℃.

The amount of compound 3c labeled [ ¹⁸ F] obtained as a result of the reaction was 7.70 mCi, and the radiation doses remaining in the reaction vessel and silica SepPak were 0.64 and 2.08 mCi, respectively (see FIGS. 1 and 2).

Step 2: 1- {2- {2- [2- (2- [ ¹⁸ F] fluoroethoxy) ethoxy] ethoxy} ethyl} -4- (N-maleimidylmethyl) -1,2, Preparation of 3-triazole ([ ¹⁸ F] 1a)

The compound 3c (7.70 mCi) labeled with [ ¹⁸ F] prepared in Step 1 of Example 4 was dissolved in acetonitrile (1.0 mL), and then 0.2 mL of the compound 4a (2 mg) prepared in Preparation Example 17 was obtained. ) Into a reaction vessel containing. 0.2 mL of an acetonitrile solution (5 mg (CuI) and 10 μl (Et ₃ N) dissolved) of CuI and triethylamine was added thereto, followed by stirring at 50 ° C. for 10 minutes and cooling to room temperature.

The labeling yield using radio thin layer chromatography was 92.3% (see FIG. 3).

Example 5 1- {2- {2- [2- (2- [ ¹⁸ F] fluoroethoxy) ethoxy] ethoxy} ethyl} -4- (3- (N-maleimidyl) propyl ) -1,2,3-triazole ([ ¹⁸ F] 1b)

[ ¹⁸ F] 3c (7.70 mCi) prepared in Step 1 of Example 4 was dissolved in acetonitrile (1.0 mL), and 0.2 mL of the reaction container containing compound 4b (2 mg) prepared in Preparation Example 18 was prepared. Put in. 0.2 mL of an acetonitrile solution (5 mg (CuI) and 10 μl (Et ₃ N) dissolved) of CuI and triethylamine was added thereto, followed by stirring at 50 ° C. for 10 minutes and cooling to room temperature.

The labeling yield using radio thin layer chromatography was 50.0% (see FIG. 4).

Example 6 1- {2- {2- [2- (2- [ ¹⁸ F] fluoroethoxy) ethoxy] ethoxy} ethyl} -4-{{2- {2- [2- ( Preparation of N-maleimyl) ethoxy] ethoxy} ethoxy} methyl} -1,2,3-triazole ([ ¹⁸ F] 1c)

[ ¹⁸ F] 3c (7.70 mCi) prepared in Step 1 of Example 4 was dissolved in acetonitrile (1.0 mL), and 0.2 mL of the resultant was used in a reaction vessel containing Compound 4c (2 mg) prepared in Preparation Example 19. Put in. 0.2 mL of an acetonitrile solution (5 mg (CuI) and 10 μl (Et ₃ N) dissolved) of CuI and triethylamine was added thereto, followed by stirring at 50 ° C. for 10 minutes and cooling to room temperature.

The labeling yield using radio thin layer chromatography was 91.4% (see FIG. 5).

< Experimental Example  1> ¹⁸ Of the present invention labeled F Maleimide  Compound stability experiment

Compound 1c labeled with [ ¹⁸ F] prepared in Example 6 was separated by HPLC, and the obtained solution was collected by passing through C-18 SepPak, eluted with acetonitrile solution (2.0 mL), and then nitrogen at 80 ° C. Blow off the solvent to remove. Add PBS buffer (7.4 pH) solution (0.5 mL) and stir slowly for about 1 hour, and then decompose the compound 1c labeled [ ¹⁸ F] using radio thin layer chromatography at 10, 30, or 60 minutes. Was measured, and the measurement results are shown in FIG.

As a result, it was confirmed that 94.5% at 10 minutes, 90.6% at 30 minutes, and 85.5% at 60 minutes remained undecomposed (see FIG. 7).

< Experimental Example  2> ¹⁸ With F Labeled  The present invention uses the compound Glutathione ¹⁸ F marker experiment

To the [ ¹⁸ F] labeling compound (0.32 mCi) acetonitrile solution (0.1 mL) obtained in Example 6, PBS buffer solution (0.4 mL) in which glutathione (1 mg) was dissolved was added, and stirred at room temperature for 20 minutes. Radio thin layer chromatography analysis showed a 97% label yield (see FIG. 6).

From this, it can be seen that by using the maleimide compound according to the present invention, radioisotopes such as ¹⁸ F can be conveniently labeled with high yield on biological compounds.

Claims (16)

Maleimide compound which has a 1,2,3-triazole group represented by following General formula (1).
[Formula 1]

(In Formula 1,
X is-[CH ₂ CH ₂ O] _n CH ₂ CH _2- ,
Y is a C ₁ -C ₆ straight or branched alkyl group; Or -CH ₂ [OCH ₂ CH ₂ ] _n- ,
n is an integer from 1 to 5,
Wherein F is ¹⁸ F or ¹⁹ F)
The method of claim 1,
X is-[CH ₂ CH ₂ O] _n CH ₂ CH _2- (n is an integer of 1-3),
Y is methyl; ethyl; n-propyl; Or -CH ₂ [OCH ₂ CH ₂ ] _n- (n is an integer from 1-4),
F is a ¹⁸ F or ¹⁹ F maleimide compound having a 1,2,3-triazole group.
According to claim 1, wherein the maleimide compound having a 1,2,3-triazole group
(1) 1- {2- {2- [2- (fluoroethoxy) ethoxy] ethoxy} ethyl} -4- (N-maleimidylmethyl) -1,2,3-triazole;
(2) 1- {2- {2- [2- (2-fluoroethoxy) ethoxy] ethoxy} ethyl} -4- (3- (N-maleimidyl) propyl) -1,2, 3-triazole;
(3) 1- {2- {2- [2- (2-fluoroethoxy) ethoxy] ethoxy} ethyl} -4-{{2- {2- [2- (N-maleimidyl) Ethoxy] ethoxy} ethoxy} methyl} -1,2,3-triazole;
(4) 1- {2- {2- [2-((fluoro-18) ethoxy) ethoxy] ethoxy} ethyl} -4- (N-maleimidylmethyl) -1,2,3- Triazoles;
(5) 1- {2- {2- [2- (2- (fluoro-18) ethoxy) ethoxy] ethoxy} ethyl} -4- (3- (N-maleimyl) propyl)- 1,2,3-triazole; And
(6) 1- {2- {2- [2- (2- (fluoro-18) ethoxy) ethoxy] ethoxy} ethyl} -4-{{2- {2- [2- (N- Maleimide) ethoxy] ethoxy} ethoxy} methyl} -1,2,3-triazole. A maleimide compound having a 1,2,3-triazole group, characterized by the above-mentioned.
As shown in Scheme 1 below,
Obtaining a fluoroalkyl azide compound represented by Chemical Formula 3 by nucleophilic fluorination reaction of the sulfonate precursor represented by Chemical Formula 2 under an organic solvent (Step 1); And
Reacting the fluoroalkyl azide compound of formula 3 obtained in step 1 with a maleimide having a terminal alkyne group represented by formula 4 under an organic solvent and a copper catalyst to obtain a compound represented by formula 1 (step 2) Method for producing a maleimide compound having a 1,2,3-triazole group of claim 1
[Reaction Scheme 1]

(In Scheme 1,
X and Y are as defined in Formula 1 of claim 1,
R is methyl, trifluoromethyl, p -toluenyl, p -nitrophenyl, soluble polyethylene glycol, polystyrene or insoluble polystyrene)
The method for producing a maleimide compound having 1,2,3-triazole group according to claim 4, wherein R is methyl, p -toluenyl or p -nitrophenyl.
The method of claim 4, wherein the solvent of step 1 is selected from the group consisting of acetonitrile, dimethylformamide (N, N-dimethylformamide, DMF), dimethylsulfoxide (dimethylsulfoxide, DMSO) and tertiary alcohol or a mixed solvent thereof. Method for producing a maleimide compound having a 1,2,3-triazole group, characterized in that any one kind.
The method for producing a maleimide compound having a 1,2,3-triazole group according to claim 6, wherein the tertiary alcohol is t -amyl alcohol.
The method of claim 4, wherein the fluoride of step 1 is tetrabutylammonium fluoride (TBAF) or cesium fluoride (CsF) of the maleimide compound having a 1,2,3-triazole group, characterized in that Manufacturing method.
The method of claim 4, wherein the solvent of step 2 is a group consisting of acetonitrile, dimethylformamide (N, N-dimethylformamide, DMF), dimethylsulfoxide (DMSO), distilled water, tertiary alcohol, and a mixed solvent thereof. Method for producing a maleimide compound having a 1,2,3-triazole group, characterized in that any one selected from.
10. The method for producing a maleimide compound having a 1,2,3-triazole group according to claim 9, wherein the organic solvent is acetonitrile.
The copper catalyst of step 2, wherein the copper catalyst of step 1 is copper iodide (CuI), copper bromide (CuBr) and copper chloride (CuCl), or any one monovalent copper compound or copper sulfate (CuSO ₄ ) or copper acetate. (Cu (OAc) ₂ ), copper nitrate (Cu (NO ₃ ) ₂ , trifluoromethsulfonate sulfide (Cu (OTf) ₂ ) and copper oxide (CuO); A method for producing a maleimide compound having a 1,2,3-triazole group, which is a valent copper compound.
12. The method of claim 11, further comprising using a reducing agent selected from the group consisting of Na-ascorbate, sodium sulfite (Na ₂ SO ₃ ), dithiothreitol when using a copper catalyst having an oxidation number of 2 The manufacturing method of the maleimide compound which has a 1,2,3-triazole group characterized by the above-mentioned.
The method of claim 12, wherein the copper catalyst is CuI or CuSO ₄ / Na-ascorbate.
5. The method according to claim 4, wherein when using a copper catalyst having an oxidation number of 1, it is selected from the group consisting of alkali metal salts of bicarbonate ions, alkali metal salts of carbonate ions, triethylamine, diisopropylethylamine, pyridine, lutidine and collidine Method for producing a maleimide compound having a 1,2,3-triazole group, characterized in that any one base used together.
15. The method for producing a maleimide compound having 1,2,3-triazole group according to claim 14, wherein the base is triethylamine or diisopropylethylamine.
A composition for labeling a biological compound having an SH-functional group containing a maleimide compound having a 1,2,3-triazole group represented by Formula 1 as an active ingredient:
[Formula 1]

(In Formula 1,
X is-[CH ₂ CH ₂ O] _n CH ₂ CH _2- ,
Y is a C ₁ -C ₆ straight or branched alkyl group; Or -CH ₂ [OCH ₂ CH ₂ ] _n- ,
n is an integer from 1 to 5,
Wherein F is ¹⁸ F).

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