KR101427292B1 - F-18 labeled triazanonane derivatives or pharmaceutically acceptable salt thereof for hypoxic tissue imaging - Google Patents

Abstract

The present invention relates to a fluoro-18 labeled triazanonane derivative or a pharmaceutically acceptable salt thereof for ischemic tissue imaging. The triazanonane derivative or a pharmaceutically acceptable salt thereof according to the present invention can stably form a complex with a metal and an octahedral structure with F-18, and can label F-18 with an excellent yield of 80-90% Can be usefully used in the manufacture of radiopharmaceuticals, and thus can be usefully used in positron tomography for determining epilepsy, cerebrovascular diseases, various cancers, cerebral neurological diseases, and coronary artery diseases.

Description

18-labeled triazanonane derivatives or their pharmaceutically acceptable salts for ischemic tissue imaging. ≪ RTI ID = 0.0 > (F-18 < / RTI > labeled triazanonane derivatives or salts thereof for hypoxic tissue imaging)

The present invention relates to a fluoro-18 labeled triazanonane derivative or a pharmaceutically acceptable salt thereof for ischemic tissue imaging.

Positron Emission Tomography (PET) provides 3-D imaging of the human body in real time using a radioactive isotope-labeled molecular probe that emits positron to provide information on the presence and progress of the disease, Is the diagnostic imaging molecular imaging technology applicable to the human body. Fluorine-18 ( ¹⁸ F) is easily produced in high purity from medical particle accelerators among the various radionuclides that emit positron. It is very useful for positron emission tomography in consideration of overall manufacturing process / time including label reaction, It is the most studied radionuclide because of its suitable characteristics.

Fluorine-18 (F-18), which is most widely used for positron emission tomography (PET), is produced in a cyclotron and is supplied in a molten state in water. Since the fluoride is fluoride, it's difficult. Therefore, a method of removing water and adding a substance such as tetrabutylammonium or crown ether to activate it and then reacting it with an organic solvent with a precursor having a leaving group is generally widely used.

In order to remove water during the production of positron-emitting radiopharmaceuticals labeled with F-18, a method of bubbling or decompressing the gas while heating is generally used. In this case, a size of 1 to 10 mL or more is required, which is disadvantageous in that the size of the automatic synthesizer can not be reduced and the time required for evaporating water is longer than other processes, which leads to a longer synthesis time. Thus, much research has been done on F-18 labeling to make processes that do not require the removal of water.

Among the most advanced methods are immunomedics patent WO 2008/088648, US 2008/0253964, US 2009/0155166 and a series of papers (McBride WJ, et al. A novel method of 18F radiolabeling for PET , Et al., J. Nucl Med (2009) 50: 991-998; Laverman P, et al. A novel facile method of labeling octreotide with 18F-fluorine. J Nucl Med (2010) 51: 454-461; McBride WJ, 18F labeling of peptides with a fluoride-aluminum-chelate. According to Bioconjug Chem (2010) 21: 1331-1340, a complex of F-18 and aluminum is first formed and then bound to a biologically active substance such as octreotide V It is disclosed that the NOA derivative is chelated to form a chelate, which is a reaction occurring in an aqueous solution. Therefore, there is no need to add a step of removing water separately, thereby simplifying the reaction process and shortening the reaction time.

However, since the F-18 labeling method by the immunomedix method does not require the step of removing water, it is possible to more economically label F-18, but the disadvantage that the labeling efficiency remains at the level of 50% There is a need to dramatically improve this.

Accordingly, the inventors of the present invention conducted studies to improve labeling efficiency, and found that a novel triazanonane derivative was synthesized and showed that the triazanonane derivative had an effect for improving the labeling efficiency of F-18, and completed the present invention Respectively.

Patent Document 1: WO 2008/088648 Patent Document 2: US 2008/0253964 Patent Document 3: US 2009/0155166

Non-Patent Document 1: McBride WJ, et al. A novel method of 18F radiolabeling for PET, J Nucl Med (2009) 50: 991-998 Non-Patent Document 2: Laverman P, et al. A novel facile method of labeling octreotide with 18F-fluorine. J Nucl Med (2010) 51: 454-461 Non-Patent Document 3: McBride WJ, et al. Improved 18F labeling of peptides with a fluoride-aluminum-chelate. Bioconjug Chem (2010) 21: 1331-1340

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

Another object of the present invention is to provide a complex of the triazanonane derivative with a metal and fluorine.

It is still another object of the present invention to provide a radiopharmaceutical comprising the triazanonane derivative, a metal and a fluorine complex as an active ingredient.

Another object of the present invention is to provide a kit for forming a complex of the triazanone derivative with aluminum and fluorine-18.

In order to achieve the above object, the present invention provides a triazanonane derivative represented by the following formula (I) or a pharmaceutically acceptable salt thereof.

[Chemical Formula 1]

(The above substituents are as described in the present specification).

The present invention also provides a complex of the triazanonane derivative with a metal and fluorine.

Further, the present invention provides a radiopharmaceutical comprising the triazanonane derivative, a metal and a fluorine complex as an active ingredient.

The present invention also provides a kit for forming a complex of the triazanonane derivative with aluminum and fluorine-18.

The triazanonane derivative or its pharmaceutically acceptable salt according to the present invention can form a stable complex with metals and F-18 and an octahedral structure and can label F-18 with an excellent yield of 80-90% It can be usefully used in the production of positron emission radioactive medicine, and thus can be usefully used for positron tomography for determining epilepsy, cerebrovascular disease, various cancers, cerebral nervous system diseases and coronary artery diseases.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a view showing data obtained by separating F-18-labeled compounds by passing them through an alumina cartridge and then separating them by TLC in an embodiment of the present invention. FIG.
2 is checking the stability in the serum at room temperature and then labeled with ¹⁸ F in the compound (a) NODA-Et-NI and (b) NODA-Pr-NI of Examples 1 and 2 of the present invention up to 4 hours Fig.
FIG. 3 is a graph showing the changes in the activity of the compounds of Examples 1 and 2 of the present invention (a) NODA-Et-NI and (b) NODA-Pr-NI after labeling ¹⁸ F with HeLa, CHO, CT26 cells under normal oxygen and hypoxic conditions And the results are shown in Fig.
Figure 4 shows the results of intravenous injection of mice injected with CT26 tumor cells after ¹⁸ F labeled with the compounds (a) NODA-Et-NI and (b) NODA-Pr-NI of Examples 1 and 2 of the present invention, Fig.
FIG. 5 shows the results of intravenous injection of mice implanted with CT26 tumor cells after ¹⁸ F of the compounds (a) NODA-Et-NI and (b) NODA-Pr-NI of Examples 1 and 2 of the present invention were labeled, Minute, 1 hour, and 2 hours after the injection.

Hereinafter, the present invention will be described in detail.

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

In Formula 1,

,

또는

이고; 및Wherein R is

,

or

ego; And

n is an integer of 1 to 3;

The triazanonane derivative of formula (1) according to the present invention is based on a mother cell (hereinafter referred to as NODA) in which an acetyl group is introduced into two nitrogen atoms of a triazanonane ring, and the other one contains various bioactive substances .

N, and S atoms capable of imparting electrons to the metal are not in a position capable of forming a pentachronous or hexagonal shape with the metal forming the complex at the center of the NODA. If O, N, S In the case where the atom exists at a position where the metal can form a complex with the center of the NODA and a pentagonal or hexagonal shape, there is a problem that the labeling efficiency is lowered due to interference with F-18 when it forms a complex with the metal, Compounds that are not affected when F-18 is labeled are being developed by forming such that the substituents in the conventional NODA parent nucleus are free of N, O, or S, or when these atoms form a complex with the metal,

In the meantime, the derivative according to the present invention is a tertiary amine and conjugated with a double bond in the nitrogen of the nitroimidazole or the nitrotriazole ring, so that the action as an electron donor is weak, so that nitrogen forms a pentagon or a hexagon But it has the advantage of high labeling efficiency.

Preferred examples of the triazanonane derivative represented by the formula (1) according to the present invention are as follows:

(1) Synthesis of 2,2 '- (7- (2- (2-nitro-1H-imidazol-1-yl) ethyl) -1,4,7-triazonan- ; or

(2) Synthesis of 2,2 '- (7- (3- (2-nitro-1H-imidazol-1-yl) propyl) -1,4,7-triazonan- .

The triazanonane derivative represented by the formula (1) of the present invention can be used in the form of a pharmaceutically acceptable salt, and the acid addition salt formed by a pharmaceutically acceptable free acid is useful as a salt. 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, tartaric acid, malonic acid, , 4-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.

In addition, the triazanonane derivative represented by Formula 1 of the present invention includes 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.

In addition, the present invention relates to a process for producing a compound represented by the formula (1)

A step of dissolving a NODA compound represented by the formula (2) protected with a protecting group and a base in an organic solvent, and then adding a nitroimidazole or a nitrotriazole compound to obtain a compound represented by the formula (3) (step 1); And

(2) a step of dissolving the compound represented by the formula (3) obtained in the above step 1 in an organic solvent and then carrying out a deprotection reaction to obtain a compound represented by the formula (1) (step 2) ≪ / RTI >

[Reaction Scheme 1]

(Wherein n and R in the above Reaction Scheme 1 are as defined in Formula 1, and P is a protecting group. Examples of the protecting group include an alkyl protecting group such as ^t butyl, a benzyloxycarbonyl group (Cbz), a t-butoxycarbonyl group (t- -Methoxybenzyl group (PMB) or 9-fluorenylmethoxycarbonyl group (Fmoc)

Hereinafter, the manufacturing method according to the present invention will be described step by step.

Step 1

In the step 1 of the present invention, the NODA compound represented by the formula (2) protected with a protecting group and a base are dissolved in an organic solvent, and the reaction is carried out by adding a nitroimidazole or a nitrotriazole compound to obtain a compound represented by the formula .

The organic solvent may be selected from the group consisting of acetonitrile, tetrahydrofuran, methylene chloride, dimethylformamide and dimethyl sulfoxide or a mixed solvent thereof, and acetonitrile and tetrahydrofuran Is preferably used.

The base can be potassium carbonate or sodium carbonate, and it is preferable to use potassium carbonate.

Specifically, the NODA compound represented by Formula 2 and potassium carbonate were dissolved in acetonitrile, and then a nitroimidazole-propyl bromide compound was added to the mixture. Acetonitrile was added dropwise to the mixture, and the mixture was stirred for 20 hours After the reaction was completed by stirring at room temperature with stirring, column chromatography was carried out to obtain the compound of formula (1).

Step 2

Step 2 according to the present invention is a step of preparing a compound represented by the formula (1) by performing a deprotection reaction of the compound represented by the formula (3) prepared in the step (1).

The compound represented by Formula 3 may be obtained by dissolving the compound represented by Formula 3 in a solvent, adding an acid, and performing a reaction.

At this time, hydrochloric acid, sulfuric acid, nitric acid, acetic acid or trifluoroacetic acid can be used as the acid which can be used, and hydrochloric acid can be preferably used.

As the usable solvent, dichloromethane, chloroform, tetrahydrofuran, diethyl ether, toluene, dimethylformamide or 1,4-dioxane which does not adversely affect the reaction can be used, and 1,4- Dioxane may not be used.

Further, the reaction temperature is not particularly limited, but can be performed within a range of room temperature to the boiling point of the solvent.

Specifically, the compound represented by Formula 3 is dissolved in 1,4-dioxane solvent and concentrated hydrochloric acid is added. After stirring at room temperature for 15 hours, RP-HPLC is performed to obtain a compound represented by Formula (1).

Furthermore, the present invention provides a complex of a triazanonane derivative represented by the formula (1), a metal and a fluorine forming an octahedron having six ligands.

The fluoride is preferably F-18.

The metal may be aluminum, gallium, indium, thallium, yttrium or ruthenium, preferably aluminum.

When aluminum in the metal is bonded to the triazanonane ring of the compound of Formula 1, the ligands of the three nitrogen atoms and the COOH in the triazanonane ring can easily bond to aluminum, and the structure in which aluminum is bonded is pentagonal Thereby forming a stable structure. In the case of aluminum, since it is a metal that binds to 6 ligands, one bond can be added, and if it forms a bond with another ligand in the same molecular structure, F-18 added for labeling the compound And it is considered that the binding ability with F-18 is excellent if it forms an unstable bond with another ligand in the same molecular structure.

In the method for producing the complex, a polymer cartridge may be used as the ¹⁸ F label. Specifically, MCl ₃ 6H ₂ O (M is a metal) was dissolved in 0.1 M sodium acetate buffer (pH 4) to make a 2 mM MCl ₃ stock solution. Then, 0.4 M KHCO ₃ and distilled water were flowed into a polymer cartridge QMA light cartridge , ¹⁸ F produced in the cyclotron was shed and collected. Physiological saline can be poured into the cartridge to elute the compound labeled with metal- ¹⁸ F, diluted by adding a reaction solvent to the solution, and passed through an Alumina N cartridge to prepare the complex.

The present invention also provides a radiopharmaceutical comprising a triazanonane derivative represented by the above formula (1), a metal and a fluoride complex as an active ingredient.

The complex forms a stable structure by forming a tetrazonan derivative and a metal and an octahedron in which F-18 has 6 ligands, thereby being able to label F-18 at 84-90%, and is useful as a radiopharmaceutical using PET or the like Lt; / RTI >

Further, the present invention provides a kit for forming a complex of the triazanonane derivative with a metal such as aluminum and fluorine-18.

The kit is a non-heat-sterilizing type F-18 labeled radioactive pharmaceutical preparation kit. To facilitate the labeling of F-18 using a metal complex compound on the triazanonane derivative, a triazanonane derivative and a suitable buffer solution are in a liquid state And the mixture is dispensed into a pharmaceutically preferable sterilization vial and sealed or stored in a refrigerator, freezer or freeze-dried, and then used when necessary.

Hereinafter, the present invention will be described in detail with reference to Examples and Experimental Examples.

However, the following examples are illustrative of the present invention, and the present invention is not limited to the following examples.

Example 1 Synthesis of 2,2 '- (7- (2- (2-nitro-1H-imidazol-1-yl) ethyl) -1,4,7-triazonan- 1,4-diyl) diacetic acid; NODA-Et (2-nitro-1H-imidazol-1-yl) ethyl) -1,4,7-triazonane- -NI)

Step 1: Preparation of 2,2 '- (7- (2- (2-nitro-1H-imidazol-1-yl) ethyl) -1,4,7- triazonan- Preparation of t-butyl esters

NODA ^t Bu (0.05 g, 0.139 mmol) and K ₂ CO ₃ (0.058 g, 0.42 mmol) represented by the above formula 4 were added to acetonitrile (2 mL), and the mixture was treated with nitroimidazolyl- After the addition of propyl bromide (0.098 g, 0.42 mmol), acetonitrile (1 mL) was added dropwise and the reaction was stirred at room temperature for 20 hours. The reaction was terminated by TLC (methylene chloride: methanol = 9: 1), filtered, and the filtrate was dried under reduced pressure. The reaction product was separated and purified by flash column chromatography (methylene chloride / methanol = 90: 10) to obtain 0.051 g (0.99 mmol) of a yellow solid in a 70% yield.

_{^{1 H NMR (CD 3 OD,}} 300 MHz): δ 7.37 (s, 1H), 4.53-4.49 (t, J = 12 Hz, 2H), 3.25 (s, 4H), 2.84-2.81 (m, 8H), 2.66-2.64 (m, 4H), 2.46-2.42 (t, 2H), 1.96-1.87 (m, 2H), 1.39 (s, 18H).

¹³ C NMR (CD ₃ OD, 75 MHz):? 171.4, 144.7, 128.2, 127.2, 80.8, 59.7,

56.1, 55.8, 55.6, 54.6, 48.2, 28.4, 28.1.

^{Mass specturm (ESI +): [} M + H] + calculated for C 23 H 41 N 6 O 6: 511.32; found 511.2.

TLC Rf = 0.4 (95% MC, 5% MeOH)

Step 2: 2,2 '- (7- (2- (2-Nitro-1H-imidazol-1-yl) ethyl) - 1,4,7-triazonan- Diacetic Acid  Produce

To the dioxane (2 mL) solution of compound NODA-tBu-Et-NI (0.026 g, 0.05 mmol) prepared in step 1 above in concentrated hydrochloric acid (0.5 mL) was added and stirred at room temperature for 15 hours The reaction was carried out. After the completion of the hydrolysis reaction was confirmed using ESI-Mass, the reaction mixture was filtered with a Whatman injection filter (0.45 μm) and dried under reduced pressure. The final product was separated by RP-HPLC (A: 10 mM HCl, B: MeCN; B concentration was increased from 0 to 100% for 30 min and B 100% was kept for 5 min.) And the recovered liquid was dried to give pale yellow 0.017 g of the aimed compound was obtained in 85% yield.

_{^{1 H NMR (CD 3 OD,}} 300 MHz): δ 7.34 (s, 1H), 6.99 (s, 1H), 3.78 (s, 4H), 3.51 (s, 2H), 3.36 (s, 4H), 3.26 ( s, 4H), 3.19 (s, 5H), 1.81 (s, 1H).

¹³ C NMR (CD ₃ OD, 75 MHz):? 172.3, 145.0, 129.0, 128.8, 57.2, 55.7, 51.2, 50.9, 45.2.

^{Mass spectrum (ESI +), [} M + H] + calculated for C 15 H 25 N 6 O 6: 385.18; Found 385.3.

^{HRMS (ESI +), (M} + H) + calculated for C 15 H 25 N 6 O 6: 385.1836; Found 385.1837.

Example 2 Synthesis of 2,2 '- (7- (3- (2-nitro-1H-imidazol-1-yl) propyl) -1,4,7-triazonan- 1,4-diyl) diacetic acid (NODA-1-yl) propyl) -1,4,7-triazonane- Et-NI)

Step 1: Preparation of 2,2 '- (7- (3- (2-nitro-1 H-imidazol-1-yl) propyl) -1,4,7- triazonan- Acid  Preparation of t butyl ester

The procedure of Step 2 of Example 2 was repeated except that ethyl-4-bromovalerate (0.030 g, 0.153 mmol) was used instead of nitroimidazolyl- To obtain the target compound (0.051 g, 70%).

¹ H NMR (CDCl ₃ , 300 MHz, 25 属 C): δ 1.19 (t, 3H, J = 6 Hz), 1.38 (s, 18H), 1.69-1.59 (m, 2H), 1.83-1.76 ), 2.31 (t, 2H, J = 6 Hz), 2.71-2.66 (m, 2H), 3.80-3.03 (m, 16H), 4.06 (q, 2H).

¹³ C NMR (CDCl ₃ , 75 MHz, 25 属 C): δ 14.1, 21.8, 23.9, 28.1, 33.3, 48.8, 52.1, 55.5, 57.6, 60.4, 81.6, 170.6 (CO), 172.8 (CO) ppm.

ESI-MS: m / z = 486.5 for [M + H] < + >.

Step 2: 2,2 ' - (7- (3- (2-nitro-lH- Imidazole -1-yl) propyl) -1,4,7- Triazonan -1,4- Dai ) Diacetic Acid  Produce

Except that NODA-tBu-Pr-NI (0.024 g, 0.05 mmol) represented by the formula 6 was used instead of the compound NODA-tBu-Et-NI represented by the formula 5 in the step 2 of Example 1 Was carried out in the same manner as above to obtain 0.015 g (0.38 mmol, 83%) of the desired compound represented by the formula (1B) as a pale yellow compound.

_{^{1 H NMR (CD 3 OD,}} 300 MHz): δ 7.27 (s, 1H), 6.96 (s, 1H), 4.34 (brs, 2H), 3.75 (brs, 5H), 3.39-3.47 (m, 5H), 3.29 (br s, 4H), 3.16 (s, 4H), 2.23 (brs, 2H).

¹³ C NMR (CD ₃ OD, 75 MHz):? 172.6, 144.8, 128.5, 128.4, 63.0, 57.2, 55.6, 51.6, 51.3, 50.3, 47.3, 25.3.

^{Mass spectrum (ESI +), [} M + H] + calculated for C 16 H 27 N 6 O 6: 399.2; Found 399.3.

^{HRMS (ESI +), (M} + H) + calculated for C 16 H 27 N 6 O 6: 399.1992; Found 399.1996.

≪ Experimental Example 1 > F-18 label

¹⁸ F was labeled on the compounds synthesized in the above examples and the following experiment was conducted to investigate the labeling efficiency.

AlCl ₃ 6H ₂ O was dissolved in 0.1 M sodium acetate buffer (pH 4) to prepare a 2 mM AlCl ₃ stock solution. The QMA light cartridge was activated by flowing 5 mL of 0.4 M KHCO ₃ and 10 mL of distilled water and collecting ¹⁸ F produced from the cyclotron. 0.4 mL of physiological saline was poured into the cartridge to elute ¹⁸ F. AlCl ₃ stock solution was added thereto and the mixture was allowed to stand at room temperature for 10 minutes to prepare an Al ¹⁸ F solution containing 27 to 90 nmol of Al. To 1 mL of 0.1 M sodium acetate buffer solution, 30-100 nM of the compound of the above Example was dissolved, and the Al ¹⁸ F solution prepared above was added thereto, followed by reaction in a heating block heated at 110 캜 for 10 minutes for labeling. Labeled compounds were purified by passing through an Alumina N cartridge.

The labeling efficiency was measured using a TLC scanner (Bioscan AR-2000) and an autoradiography (BAS2500 Imaging System) after developing 80% MeCN with Instant Thin Layer Chromatography Silica Gel (ITLC-SG, Gelman Science, Ann Arbor, MI) Were detected and calculated. After labeling, the unlabeled ¹⁸ F remained at the origin when identified by ITLC, and the labeled compounds were identified by TLC scanner to have Rf between 0.4 and 0.5, which was purified through Alumina N cartridge ¹ F can be completely removed (see FIG. 1).

The labeling efficiencies for the compounds of the Examples are shown in Table 1 below.

compound Label Efficiency (%) Example 1 84.5 Example 2 87.9

As shown in Table 1, it was confirmed that the triazanone derivatives according to the present invention all exhibited a good labeling efficiency of 84% or more.

Therefore, it can be usefully used in the production of positron-emitting radiopharmaceuticals, and thus can be usefully used for positron tomography for judging various cancers, cerebral neurological diseases, coronary artery diseases and the like.

≪ Experimental Example 2 >

The following experiment was conducted to examine the stability at room temperature and the stability in serum at 37 캜 for a solution in which the ¹⁸ F-labeled compound of the present invention was dissolved.

The labeled compounds were incubated for 10 minutes, 30 minutes, 60 minutes, 120 minutes and 240 minutes under the above conditions, and the degree of decomposition of the compounds was developed with 80% acetonitrile using ITLC. The results are shown in FIG. 2 .

As a result, as shown in Fig. 2, it was confirmed that the compounds of Examples 1 and 2 were very stable for 4 hours.

Therefore, it can be usefully used in the production of positron emitting radiopharmaceuticals, and thus can be usefully used for positron tomography for determining epilepsy, cerebrovascular diseases, various cancers, cerebral neurological diseases and coronary artery diseases.

<Experimental Example 3> Measurement of serum protein binding

The following experiment was conducted to determine whether the ⁶⁸ Ga-labeled compound according to the present invention is bound to phosphorus serum protein.

The ⁶⁸ Ga labeled compounds were each incubated with serum (1 mL) at 37 ° C. for 10 minutes and 60 minutes, and then loaded on a PD-10 column (pre-conditioned with 1 mL of 1% BSA / 0.1 M DTPA) PBS, and received in 30 test tubes in 0.5 mL fractions. The radioactivity of each test tube was measured with a gamma counter. Samples were taken from each test tube in 2 L increments, taken on a filter paper, and stained with Coomassie blue to identify protein peaks. The radioactive curve of each fraction was plotted to calculate protein binding percentages. The results are shown in Table 2.

Protein binding (%) 10 minutes 60 minutes ⁶⁸ Ga-NODA-Et-NI
(Example 1) 0.24 + 0.03 0.64 + 0.03 ⁶⁸ Ga-NODA-Pr-NI
(Example 2) 0.32 ± 0.06 0.36 + 0.07

As shown in the above Table 2, it was predicted that the binding rate with serum proteins of both of the compounds of Examples 1 and 2 labeled with ⁶⁸ Ga was so low that the loss rate in blood was excellent.

Therefore, it can be usefully used in the production of positron emitting radiopharmaceuticals, and thus can be usefully used for positron tomography for determining epilepsy, cerebrovascular diseases, various cancers, cerebral neurological diseases and coronary artery diseases.

 <Experimental Example 4> In vitro test

The following experiment was conducted to determine whether the labeled compounds according to the present invention were ingested in vitro. At this time, CHO (Chinese Hamster Ovarian cancer), Hela (Henrietta Lacks cervical cancer) and CT26 (mouse colon cancer) were used as the cell lines.

First, all cell lines were cultured in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum and 1% antibiotic mixture at 37 ° C in air supplemented with 5% carbon dioxide. For the experiment, 4x10 ⁵ cells were cultured in a 12-well plate overnight, and then cultured for 4 hours under normoxia (air containing 5% CO ₂ ) and hypoxia (nitrogen containing 5% CO ₂ ) for 4 hours. 100 μL (0.18 MBq) of compound NODA-Et-NI of Example 1 labeled with F-18 and compound NODA-Pr-NI of Example 2 were added thereto and further incubated for 60 minutes.

At 5, 15, 30, and 60 minutes, the wells were washed with DMEM, 0.5% SDS solution was added, and the cells were lysed and measured with a gamma counter to measure the radioactivity in the cells. The amount of protein in the dissolved cell fluid was measured by a bactic tonic acid method and the amount of radioactivity ingested per mg protein was calculated.

As a result, as shown in FIG. 3, both compounds of the examples according to the present invention showed increased cell uptake under hypoxic conditions in all cell lines, and showed that they could be used for imaging of hypoxic cancer cells (see FIG. 3).

Therefore, it can be usefully used in the production of positron emitting radiopharmaceuticals, and thus can be usefully used for positron tomography for determining epilepsy, cerebrovascular diseases, various cancers, cerebral neurological diseases and coronary artery diseases.

&Lt; Experimental Example 5 >

In order to confirm the mouse in vivo distribution of the labeled compound according to the present invention, the following experiment was conducted.

First, CT26 cells were cultured in RPMI1640 supplemented with 10% fetal bovine serum and 1% antibiotic solution. The cells were then washed with 10 mL of PBS and injected with 4 × 10 ⁵ cells / mL on balb / c mice for 14 days, Of tumors were prepared. 0.15 MBq / 0.1 mL of the nitroimidazole-triazanonane derivative labeled with F-18 was injected through the tail vein for in vivo distribution test.

At 10, 60 and 120 minutes, mice were sacrificed and tissues such as tumor, blood, and muscle were excised, weighed, and radioactivity was measured. The radioactivity ingested into each tissue is shown in FIG. 4 as a percentage (% ID / g) taken per gram of tissue versus the administered dose.

As a result, as shown in FIG. 4, both compounds of the examples according to the present invention had the highest kidney intake at 11.1% ID / g and 9.8% ID / g at 10 minutes, which rapidly decreased over time . Followed by liver ingestion, which slowed down over time. Blood and lung intake was high at 10 minutes, but decreased rapidly. The tumor / muscle uptake rate was very high at NODA-Et-NI = 14.5 after 1 hour and NODA-Pr-NI = 37.44 after 2 hours and was shown to be useful for hypoxic cancer tissue imaging (see FIG. 4).

Therefore, it can be usefully used in the production of positron emitting radiopharmaceuticals, and thus can be usefully used for positron tomography for determining epilepsy, cerebrovascular diseases, various cancers, cerebral neurological diseases and coronary artery diseases.

Experimental Example 6: Mouse PET image test

The following PET imaging experiments were performed to determine the in vivo distribution of the labeled compounds according to the present invention.

14.4 MBq (0.1 mL) of NODA-Et-NI and NODA-Pr-NI labeled with F-18 were injected into the tail vein of mice transplanted with CT26 tumor as in Experimental Example 5, and injected with 2% isoflurane PET images were taken using an animal PET / CT scanner (GE Healthcare, Princeton, NJ) at 30, 60 and 120 min after anesthesia.

As a result, as shown in FIG. 5, it was confirmed that all the compounds of the examples according to the present invention were rapidly excreted into the kidney and ingested into hypoxic cancer, and NODA-Et-NI was found to have a higher liver and kidney intake 5).

Therefore, it can be usefully used in the production of positron emission radiopharmaceuticals, and thus can be usefully used in positron tomography for determining the cerebral nervous system diseases such as epilepsy, cerebrovascular disease, Alzheimer's disease, and coronary artery disease and cardiomyopathy have.

Claims (7)

A triazanone derivative for F-18 labeling represented by the following formula (1) or a pharmaceutically acceptable salt thereof:
[Chemical Formula 1]

(In the formula 1,
Wherein R is

,

or

ego; And
and n is an integer of 1 to 3).
A compound represented by formula (1) of claim 1; Any one metal selected from the group consisting of aluminum, gallium, indium, thallium, yttrium and ruthenium, and a complex of F-18 fluoride.
delete delete The complex according to claim 2, wherein the metal is aluminum.
A radiopharmaceutical comprising a compound represented by the general formula (1) of claim 1, a metal and an F-18 complex as an active ingredient.
A kit for forming a metal and F-18 complex with a compound represented by formula (1) of claim 1.

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