KR20180083286A - Radioactive probe for detecting hydrogen sulfide - Google Patents

Abstract

The present invention relates to a probe for detecting hydrogen sulfide in vivo, and more specifically, to a probe for detecting hydrogen sulfide including a complex compound to which a radioactive isotope Cu is introduced. According to various embodiments of the present invention, as a result of using an animal model in which a large amount of hydrogen sulfide participating in various diseases is generated to carry out real-time observation through optical and nuclear medicine imaging, the probe for detecting hydrogen sulfide is selectively bound to hydrogen sulfide to selectively image a portion where hydrogen sulfide is abnormally increased in cells or tissue. Thus, the probe does not affect the anatomic characteristics of a body portion to allow discovery of a disease at a non-predictable part. In addition, the probe for detecting hydrogen sulfide has a high reaction rate with hydrogen sulfide to resolve a problem of carrying out a test after a prescribed time elapses after administration of a conventional imaging agent. Therefore, the probe can be effectively used as a disease diagnosis means such as an imaging composition or an imaging method.

Description

[0001] The present invention relates to a radioactive probe for detecting hydrogen sulfide,

TECHNICAL FIELD The present invention relates to a probe for detecting a hydrogen sulfide in vivo, and more particularly, to a probe for detecting a hydrogen sulfide containing a complex in which a radioisotope Cu is introduced.

Recent research has shown that hydrogen sulfide (H ₂ S) participates in a variety of physiological phenomena, including nitric oxide and carbon monoxide, as well as third-degree gas transporters and gasomediators It is known.

The concentration of hydrogen sulfide in blood plasma is known to be in the range of 50-100 μM. Hydrogen sulfide acts as an important signal transmitter for various physiological actions such as various inflammatory reactions, cardiovascular diseases, vasodilation, glucose metabolism and neovascularization And it is known that it is possible to diagnose diseases such as Down syndrome, Alzheimer's disease, diabetes and cirrhosis by detecting the concentration of hydrogen sulfide.

In particular, the hydrogen sulfide acts as an initiator of the K-ATP channel, contributing to the homeostasis of the cardiovascular system, and has been regarded as a healing role of damaged cardiovascular muscles. In this regard, WO204027820 A1 discloses that the hydrogen sulfide concentration is detected A real-time bio-signal measuring device for cardiac ischemia and reperfusion with a hydrogen sulfide sensor is disclosed.

In vivo hydrogen sulfide is produced through three enzymes: cystathionine-synthase (CBS), cystathionine-lyase (CSE), and 3-mercaptopurine sulpertransferase (3-MST) (GSH), which protects cells by the cellular immune system and antioxidant activity, inhibits intracellular oxidation. The intracellular hydrogen sulfide produced by CSE inhibits K _atp channel , And protects the cells by inducing GSH together with the hydrogen sulfide present in the mitochondria produced by 3-MST and CAT enzymes.

In this way, hydrogen sulfide induces changes in the concentration of GSH, thereby preventing cell deterioration by virtue of cell protection and inhibiting apoptosis. Therefore, if the detection and concentration of hydrogen sulfide in mitochondria is known, It will contribute to the study.

Accordingly, various techniques for detecting and quantifying hydrogen sulfide are being developed in a competitive manner, and development of a detection method using non-invasive images of hydrogen sulfide in the body is becoming important.

However, in order to detect and accurately measure hydrogen sulfide, it is necessary to be able to measure H ₂ S in a liquid state, and to select H ₂ S from other anion species And it is known that it is very difficult and complicated because conditions such as reduced glutathione (GSH) and selectivity are required.

In this regard, up to now, a method using a chromophore that utilizes most of intracellular cytosol and chemical properties of hydrogen sulfide in a plasma of blood, a method of detecting through a specific electrode for sulfide ion, a detection method using gas chromatography In recent years, various fluorescent probes have been developed. However, due to limitation of fluorescence probe method, detection of hydrogen sulfide through image is very limited at a small animal level. Therefore, Research on life phenomena is extremely limited.

The 2,4,6-triarylpyridinium cation compound has been disclosed as a fluorescence probe for conventional H ₂ S detection (Journal of the American Chemical Society, 125, 9000, 2003) (Analytica Chimica Acta, 631, 91, 2009). However, a method of using 2,4-dinitrobenzenesulfonylfluorosane as a fluorescence-enhanced probe has also been proposed (Analytica Chimica Acta, 631, Is hydrolyzed to change the fluorescence intensity.

In addition, an aminofluorescein compound (DPA-4-AF) having a 2,2-dipicolylamine group containing a ^divalent copper ion (Cu ^{2 +} ) has been disclosed (Myung Gil Choi, Chem. Commun . , (47), 7390-7392, 2009), WO2012-144654A1 discloses a compound in which a fluorescent substance is attached to cyclin, cyclam and TACN, When a hydrogen sulfide is reacted with the complex compound, the copper complex is released in CuS form. In this case, the copper complex is formed in the form of CuS, , And a method of quantifying the amount of hydrogen sulfide using the principle that a difference in degree of fluorescence restored according to the degree is generated. However, there is still a problem in the above-mentioned compounds until the detection of hydrogen sulfide selectively at an animal level at the level of acquiring an image at the cell level, and development of a new technique capable of replacing the conventional detection method is still required.

A problem to be solved by the present invention is to provide a radioactive probe capable of selectively detecting only hydrogen sulfide in and / or out of a living body.

According to a representative aspect of the present invention, there is provided a probe for detecting hydrogen sulfide (H ₂ S) comprising a complex in which a radioisotope Cu represented by any one of Chemical Formulas 1 to 4 is introduced:

[Chemical Formula 1]

(Wherein A, B, X ¹ to X ³ , Y ¹ , Y ² , R ¹ to R ⁸ are as defined in the present invention)

(2)

(Wherein K, L, M, R ¹⁵ to R ¹⁷ are as defined herein)

(3)

(In the above formula (3), X is as described herein.)

[Chemical Formula 4]

.

.

According to various embodiments of the present invention, the hydrogen sulfide-detecting probe according to the present invention can be used as a probe for detecting hydrogen sulfide selectively in addition to hydrogen sulfide, as a result of observing in real time through optical and nuclear medicine images using an animal model in which hydrogen sulfide is involved in various diseases. It is possible to selectively display a site where hydrogen sulfide is abnormally increased in cells or tissues so that it does not affect the anatomical characteristics of the body part and thus it is possible not only to find an unexpected part of the disease, Since the probe for detecting hydrogen sulfide has a high reaction rate with hydrogen sulfide, it can solve the problem of inspecting after a predetermined time after administration of the conventional imaging agent, and thus can be usefully used as a diagnostic tool for diseases such as a composition for imaging or a method of imaging.

1 is a graph showing the sensitivity of ⁶⁴ Cu-labeled complexes to hydrogen sulfide according to an embodiment of the present invention.
Figures 2a and 2b show the results of radio-TLC measurements of a ⁶⁴ Cu-labeled complex according to one embodiment of the invention.
3 is a graph showing the reactivity of ⁶⁴ Cu-labeled complexes according to the present invention with hydrogen sulfide over time.
FIG. 4 is a photograph showing the selective imaging of ⁶⁴ Cu-labeled complexes according to the present invention through an optical image after injection into a rat and the like.
Figures 5a-5d show image measurement results for a foot inflammation model injected with ⁶⁴ Cu-labeled complexes of Example 1 according to one embodiment of the present invention.
Figures 6a-6c show image measurement results for a foot inflammation model injected with ⁶⁴ Cu-labeled complexes of Example 20 according to one embodiment of the present invention.
FIGS. 7A to 7D show image measurement results for a foot inflammation model injected with ⁶⁴ Cu-labeled complexes of Example 2 and Example 14 according to an embodiment of the present invention. FIG.
Figures 8a through 8e show imaging measurements of foot inflammation models injected with ⁶⁴ Cu-labeled complexes of Example 3 and Example 21, according to one embodiment of the present invention.
Figures 9a and 9b show image measurement results for a foot inflammation model injected with a ⁶⁴ Cu-labeled complex of Example 7 according to an embodiment of the present invention.
FIGS. 10A and 10B show image measurement results for a foot inflammation model injected with a ⁶⁴ Cu-labeled complex of Example 9 according to an embodiment of the present invention. FIG.
FIGS. 11A-11D show image measurement results for a foot inflammation model injected with a ⁶⁴ Cu-labeled complex of Example 16 according to an embodiment of the present invention. FIG.
FIG. 12 is a graph showing the degree of inflammation of a joint region of an animal model of arthritis according to an embodiment of the present invention. FIG.
Figures 13a and 13b show image measurement results for an arthritis model in which ⁶⁴ Cu-labeled complexes of Example 1 were injected according to one embodiment of the present invention.
Figure 14 is a diagram showing image measurement results for a brain inflammation model injected with a ⁶⁴ Cu-labeled complex of Example 17 according to one embodiment of the present invention.
FIG. 15 is a diagram showing an image measurement result of a myocardial infarction model injected with ⁶⁴ Cu-labeled complex compound of Example 1 according to an embodiment of the present invention. FIG.
FIG. 16 is a diagram showing image measurement results of a myocardial infarction model injected with ⁶⁴ Cu-labeled complex compound of Example 20 according to an embodiment of the present invention. FIG.
FIG. 17 is a diagram showing an image measurement result of a myocardial infarction model injected with ⁶⁴ Cu-labeled complex compound of Example 16 according to an embodiment of the present invention. FIG.
FIG. 18 is a diagram showing image measurement results for a brain tumor model injected with ⁶⁴ Cu-labeled complex compound of Example 17 according to an embodiment of the present invention. FIG.
Figures 19a and 19b are graphs showing the results of imaging tumor (EMT6) 19a and tumor (CT26) 19b by injecting ⁶⁴ Cu-labeled complexes of Example 1, respectively, according to one embodiment of the present invention .
Figure 20 is a diagram showing the results of imaging the pancreatitis model by injecting ⁶⁴ Cu-labeled complexes of Example 1 and Example 20 according to the present invention.
Figure 21 is a diagram showing the results of imaging the septicemia model by injecting ⁶⁴ Cu-labeled complexes of Example 20 according to the present invention.
Figure 22 is a PET image shows that the hydrogen sulfide is observed in Example 1, ⁶⁴ Cu- by scanning the labeled complex high on the tongue portions in the normal rats concentration according to the invention, labeled Cu- ⁶⁴ of the first embodiment according to the invention Is capable of easily diagnosing a high concentration of hydrogen sulfide in vivo.
FIG. 23 is a graph illustrating the quantification of the degree of desorption by reacting ⁶⁴ Cu-labeled complexes of Example 20 according to the present invention with hydrogen sulfide at different concentrations. The ⁶⁴ Cu- Lt; RTI ID = 0.0 > hydrogen sulphide. ≪ / RTI >
FIG. 24 is a graph showing the concentration of hydrogen sulfide in plasma of normal rats and myocardial infarction model rats using ⁶⁴ Cu-labeled complexes of Example 20 according to the present invention, wherein ⁶⁴ Cu- The labeled complexes show very similar measurements to the conventional methylene blue method and show that the concentration of hydrogen sulphide in the plasma is significantly higher in the myocardial infarction model than in the normal rats.

Hereinafter, various aspects and various embodiments of the present invention will be described in more detail.

As used herein, the term " probe " is defined as a material capable of detecting or imaging an in vivo / external target. Generally, they are replaced with terms such as imaging agents, contrast agents, and radiopharmaceuticals.

According to one aspect of the present invention, there is provided a probe for detecting hydrogen sulfide (H ₂ S), which comprises a complex having a radioisotope Cu introduced therein represented by any one of the following Chemical Formulas 1 to 4:

[Formula 1]

(In the formula 1,

X ¹ , X ² and X ³ are each independently at least one selected from N, O and S;

Y ¹ and Y ² are each independently C or N;

Wherein A and B are each independently a ^{non-binding, C (R 9) (R} 10) -C (R 11) (R 12) -, = C (R 9) -C (R 10) (R 11) - ^{C (R 12) (R 13} ) -, C (R 9) (R 10) -C (R 11) (R 12) -C (R 13) (R 14) - , and = C (R ⁹⁾ -C (R < ¹⁰ >)=;

Each of R ¹ to R ¹⁴ is independently selected from the group consisting of a non-bond, hydrogen, hydroxy, substituted or unsubstituted C ₁ -C ₆ linear or branched alkyl, substituted or unsubstituted C ₁ -C ₆ linear or branched alkyloxycarbonyl , Substituted or unsubstituted C ₅ -C ₁₂ aryl C ₁ -C ₆ alkyl, substituted or unsubstituted C ₅ -C ₁₂ heterocycloalkyl, substituted or unsubstituted C ₅ -C ₂₀ aryl, substituted or unsubstituted C ₅ -C ₂₀ arylsulfonyl, a least one C ₁ -C ₆ alkylamine, C ₁ -C ₆ selected from dialkyl amine, a substituted or unsubstituted amine,

Said substituent is selected from the group consisting of hydroxy, hydroxy C ₁ -C ₆ alkyl, C ₁ -C ₆ dialkylamine, nitro, ⁶⁴ Cu label-3-methyl-1,4,7,10-tetraazacyclotridecane ≪ / RTI > and < RTI ID = 0.0 >

Wherein R ⁵ and R ⁸ or R ⁶ and R ⁷ may combine with each other to form a C ₄ -C ₈ cycloalkyl ring;

는 단일결합 또는 이중결합이고,remind

Is a single bond or a double bond,

The Cu is any one selected from ⁶⁰ Cu, ⁶¹ Cu, ⁶² Cu, ⁶⁴ Cu and ⁶⁷ Cu);

(2)

(In the formula (2)

Wherein K, L and N are each independently ^{C (R 18) (R 19} ) -C (R 20) (R 21) -, = C (R 18) -C (R 19) (R 20) -C ( ^{R 21) (R 22) -} , C (R 18) (R 19) -C (R 20) (R 21) -C (R 22) (R 23) - , and ^{= C (R 18) -C (} R ¹⁹ ) = < / RTI >

Each of R ¹⁵ to R ²³ is independently a bond, hydrogen, hydroxy, substituted or unsubstituted C ₁ -C ₂₀ linear or branched alkyl, substituted or unsubstituted C ₁ -C ₆ straight or branched alkyloxycarbonyl , Substituted or unsubstituted C ₅ -C ₁₂ aryl C ₁ -C ₆ alkyl, substituted or unsubstituted C ₅ -C ₁₂ heterocycloalkyl, substituted or unsubstituted C ₅ -C ₂₀ aryl, substituted or unsubstituted C ₅ -C ₂₀ arylsulfonyl, a least one C ₁ -C ₆ alkylamine, C ₁ -C ₆ selected from dialkyl amine, a substituted or unsubstituted amine,

Said substituent is selected from the group consisting of hydroxy, hydroxy C ₁ -C ₆ alkyl, C ₁ -C ₆ dialkylamine, nitro, ⁶⁴ Cu label-3-methyl-1,4,7,10-tetraazacyclotridecane ≪ RTI ID = 0.0 > 1, < / RTI >

The Cu is any one selected from ⁶⁰ Cu, ⁶¹ Cu, ⁶² Cu, ⁶⁴ Cu and ⁶⁷ Cu);

(3)

(Wherein X is hydrogen or a compound represented by the formula below).

;

;

[Chemical Formula 4]

.

.

Preferably, the complex of Formula 1 is a complex represented by Formula 5 or 6:

[Chemical Formula 5]

(In the above formula (5)

Wherein A or B is ^{C (R 32) (R 33} ) -C (R 34) (R 35) -, = C (R 32) -C (R 33) (R 34) -C (R 35) (R It represents at least one element selected from, - ³⁶⁾ and ^{C (R 32) (R 33} ) -C (R 34) (R 35) -C (R 36) (R 37)

Each of R ²⁴ to R ³⁷ is independently a bond, hydrogen, hydroxy, C ₁ -C ₆ linear or branched alkyl, C ₁ -C ₆ straight or branched alkyloxycarbonyl, C ₁ -C ₆ straight or branched chain alkyl amine, substituted or unsubstituted amine, ⁶⁴ Cu labeled 3-methyl-1, 4, 7, 10-tetraaza bicyclo in a tree in decane or nitro substituted or unsubstituted C ₅ -C ₁₂ aryl C ₁ -C ₆ C ₅ -C ₂₀ heterocycloalkyl which is unsubstituted or substituted by alkyl, hydroxy or hydroxy C ₁ -C ₆ alkyl, C ₅ -C ₂₀ arylsulfonyl which is unsubstituted or substituted by C ₁ -C ₆ dialkylamines , ≪ / RTI >

Wherein R ²⁶ and R ²⁷ or R ³⁰ and R ³¹ may combine with each other to form a C ₄ -C ₈ cycloalkyl ring;

The Cu is any one selected from ⁶⁰ Cu, ⁶¹ Cu, ⁶² Cu, ⁶⁴ Cu and ⁶⁷ Cu);

[Chemical Formula 6]

(6)

And A is ^{C (R 30) (R 31} ) -C (R 32) (R 33) - or ^{= C (R 30) -C (} R 31) = , and

Each of R ²⁴ to R ³³ is independently selected from the group consisting of a non-bond, hydrogen, hydroxy, C ₁ -C ₆ linear or branched alkyl, C ₁ -C ₆ alkylamine, C ₁ -C ₆ dialkylamine, Amine, and the like,

는 단일결합 또는 이중결합이고,remind

Is a single bond or a double bond,

가 이중결합인 경우, 상기 R²⁴ 또는 R²⁹는 비결합이며,remind

Is a double bond, R < ²⁴ > or R < ²⁹ >

The Cu is any one selected from ⁶⁰ Cu, ⁶¹ Cu, ⁶² Cu, ⁶⁴ Cu and ⁶⁷ Cu).

More preferably, the compound represented by any one of the above formulas (1) to (4)

(1) 1,4,7,10-tetraazacyclododecane;

(2) 1- (Anthracen-9-ylmethyl) -1,4,7,10-tetraazacyclododecane;

(3) 1,7-dioxa-4,10-diazacyclododecane;

(4) 1,4,7,10-tetrakis (anthracene-9-ylmethyl) -1,4,7,10-tetraazacyclododecane;

(5) (7S, 14R) -5,5,7,12,12,14-hexahexamethyl-1,4,8,11-tetraazacyclotetradecane;

(6) 2,5,5,7,9,12,12,14-octamethyl-1,4,8,11-tetraazacyclotetradecane;

(7) (1E, 7E) -2,5,5,7,9,12,12,14-octamethyl-1,4,8,11-tetraazacyclotetradeca-7,14-diene;

(8) (7S, 14S) -5,5,7,12,12,14-hexamethyl-1,4,8,11-tetraazacyclotetradecane;

(9) 6,15-Dimethyldocosu hydrodibenzo [b, i] [1,4,8,11] tetraazacyclotetradecyne;

(10) ethyl 3- (1,4,7,10-tetraazacyclododecane-1-yl) propanonate;

(11) 1,4-bis ((1,4,7,10-tetraazacyclodridecan-4-yl) methyl) benzene;

(12) 2- (4-nitrobenzyl) -1,4,7,10-tetraazacyclododecane;

(13) 1,4,7,10-tetraazacyclotridecane;

(14) Synthesis of (2R, 3S, 4S, 5R, 6S) -2- (hydroxymethyl) -6- (1,4,7,10- tetraazacyclododecan- 1- yl) tetrahydro- -3,4,5-triol;

(15) 5 - ((1,4,7,10-tetraazacyclododecan-1-yl) sulfonyl) -N, N-dimethylnaphthalen-1-amine;

(16) 1,4,8,11-tetraazacyclotetradecane;

(17) Synthesis of (2R, 2'R, 3S, 3'S, 4S, 4'S, 5R, 5'R, 6R, 6'R) -6,6 '- (1,4,7,10-tetraazacyclododecane -1,7-diyl) bis (2- (hydroxymethyl) tetrahydro-2H-pyran-3,4,5-triol);

(18) 1,4,7-triazonane;

(19) 1,5,9-triazacyclododecane;

(20) 5 - ((1,4,7-triazolon-1-yl) sulfonyl) -N, N-dimethylnaphthalen-1-amine;

(21) 1-methyl-1,4,8,11-tetraazacyclotetradecane;

(22) 1,4,8,11-tetraaza-bicyclo [6.6.3] heptadecane;

(23) bis (pyridin-2-ylmethyl) amine;

(24) N ¹ , N ¹ , N ² , N ² -tetrakis (pyridin-2-ylmethyl) ethane-1,2-diamine;

(25) Synthesis of (1Z, N'E) -N '- ((E) -3 - ((Bis (methylamino) methylene) hydrazano) butan- Osan;

(26) Synthesis of (1Z, N'E) -N '- ((E) -2 - ((bis (methylamino) methylene) hydrazano) propylidene) -N-methylcarbamohydrazonothioic acid;

(27) (2E, 2'E, 3R, 3'R) -3 - ((3 - (((R, E) -3- (Hydroxyimino) butan- -Dimethylpropyl) amino) butan-2-one oxime;

(28) 1,4,7-trithia-10-azacyclododecane;

(29) 1-methyl-1,4,7-triazonan;

(30) 1- (Anthracen-9-ylmethyl) -1,4,7-triazonan;

(31) 1-hexadecyl-1,4,7-triazonan;

(32) 1,4,7-trimethyl-1,4,7-triazonan;

(33) 1,4,7-Tris (anthracen-9-ylmethyl) -1,4,7-triazonan; And

(34) 1,4,7-trihexadecyl-1,4,7-triazonan.

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중에서 선택되는 어느 하나인 것을 특징으로 한다.The compounds represented by any one of formulas (1) to (4)

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And

And the like.

According to one embodiment of the present invention, the complexes into which the radioisotope Cu represented by any one of Chemical Formulas 1 to 4 is introduced can be used for optical and nuclear medicine images using an animal model in which hydrogen sulfide is involved in various diseases It was confirmed that the probe for detecting a hydrogen sulfide according to the present invention selectively binds to hydrogen sulfide and that the reaction rate with hydrogen sulfide is fast. Therefore, it is possible to selectively image a site where hydrogen sulfide is abnormally increased in cells or tissues And it can solve the problem of not having an unexpected part because it has no influence on the anatomical characteristics of the body part, emission tomography, gamma camera, SPECT (single photon emission computed a diagnostic imaging method such as a tomography, a Cherenkov optical image contrast agent, a contrast agent for a CCD (charge-coupled device), a contrast agent for MRI (magnetic resonance imaging), a CT (computed tomography) . ≪ / RTI >

At this time, it is preferable that the complex compound of any one of formulas (1) to (4) includes 50 μCi-1000 μCi / kg immediately before use.

Outside of this range, the signal-to-noise ratio may be excessively low, or excessive radiation dose may be increased.

According to another aspect of the present invention, there is provided a method for imaging sulfide ions in a cell or tissue using a probe for detecting hydrogen sulfide comprising a complex in which the radioisotope Cu is introduced.

In one embodiment of the present invention, the imaging of the sulfide ions is performed by measuring Cherenkov light coming from the radioactive isotope Cu.

In another embodiment of the present invention, the Cherenkov light is characterized by using a wavelength in the range of 200 to 1000 nm.

In another embodiment of the present invention, the probe for detecting a hydrogen sulfide comprising a complex having the radioisotope Cu introduced therein is a probe for detecting a hydrogen sulfide in a cell and an extracellular matrix located locally at positions or positions abnormally increased in hydrogen sulfide And is characterized by imaging sulfide ions.

According to another aspect of the present invention, there is provided a method for detecting a hydrogen sulfide comprising the steps of: providing a probe for detecting hydrogen sulfide in a pharmacological carrier, wherein the probe comprises a complex containing the radioisotope Cu; Injecting the probe into a mammal other than a human; And scanning a mammal other than a human using a radiological diagnostic imaging apparatus.

According to another aspect of the present invention, there is provided a pharmaceutical composition for diagnosing a pharmaceutically acceptable inflammatory disease comprising a probe for detecting a hydrogen sulfide comprising a complex in which the radioisotope Cu is introduced.

In one embodiment of the invention the inflammatory disease is selected from the group consisting of rheumatoid arthritis, non-rheumatoid inflammatory arthritis, arthritis associated with Lyme disease, inflammatory osteoarthritis, meningitis, osteomyelitis, inflammatory bowel disease, appendicitis, pancreatitis, Inflammatory diseases caused by inflammation, inflammation caused by inflammation, inflammation due to bacterial infection, and the like.

According to an embodiment of the present invention, an optical and nuclear medicine imaging study was conducted using an animal model that induced foot inflammation, muscle inflammation, arthritis, and brain inflammation. As a result, it was found that the radioisotope Cu according to the present invention Complex compounds can be usefully used as a pharmaceutical composition for the diagnosis of inflammatory diseases, since selective intake is confirmed at sites where inflammation is induced.

According to another aspect of the present invention, there is provided a pharmaceutical composition for diagnosing a pharmaceutically acceptable heart disease comprising a probe for detecting hydrogen sulfide comprising a complex in which the radioisotope Cu is introduced.

In one embodiment of the present invention, the heart disease is characterized by being at least one selected from myocardial infarction, cardiac ischemia, angina pectoris, cardiomyopathy, and endocarditis.

It is known that the concentration of hydrogen sulfide is high in the myocardial infarction area. The mice that have induced myocardial infarction by blocking the coronary artery of the heart of the rats are injected with the complex labeled with ⁶⁴ Cu according to the present invention, As a result of observing the degree of ingestion through the image, when a complex containing a radioisotope Cu was injected according to an embodiment of the present invention, the myocardial infarction site in the image was elevated in the myocardial infarction area so as to be clearly observed It has been confirmed that when FDG, which is a imaging agent, is injected, it is confirmed that a part of the myocardium appears as a broken ring in the image, and thus it can be usefully used as a pharmaceutical composition for diagnosing heart disease.

According to another aspect of the present invention, there is provided a pharmaceutical composition for diagnosing Parkinson's disease, which comprises a probe for detecting hydrogen sulfide comprising a complex in which the radioisotope Cu is introduced.

According to another aspect of the present invention, there is provided a pharmaceutical composition for diagnosing pharmaceutically acceptable Alzheimer's disease comprising a probe for detecting hydrogen sulfide comprising a complex in which the radioisotope Cu is introduced.

According to another aspect of the present invention, there is provided a pharmaceutical composition for diagnosing a pharmaceutically acceptable Down syndrome comprising a probe for detecting hydrogen sulfide comprising a complex in which the radioisotope Cu is introduced.

According to another aspect of the present invention, there is provided a pharmaceutically acceptable tumor diagnostic pharmaceutical composition comprising a probe for detecting hydrogen sulfide comprising a complex in which the radioisotope Cu is introduced.

According to one embodiment of the invention, the tumor diagnosis may be a hypoxia diagnosis in a tumor.

 According to another aspect of the present invention, there is provided a pharmaceutical composition for diagnosing a pharmaceutically acceptable sepsis comprising a probe for detecting hydrogen sulfide comprising a complex in which the radioisotope Cu is introduced.

According to another aspect of the present invention, there is provided a pharmaceutical composition for diagnosing pain comprising a probe for detecting hydrogen sulfide comprising a complex in which the radioisotope Cu is introduced.

According to another aspect of the present invention, there is provided a pharmaceutical composition for the diagnosis of atherosclerosis, which comprises a probe for detecting hydrogen sulfide comprising a complex in which the radioisotope Cu is introduced.

According to another aspect of the present invention, there is provided a pharmaceutical composition for diagnostic of diabetes mellitus comprising a probe for detecting hydrogen sulfide comprising a complex in which the radioisotope Cu is introduced.

According to still another aspect of the present invention, there is provided a pharmaceutical composition for diagnostic of stroke, comprising a probe for detecting hydrogen sulfide comprising a complex in which the radioisotope Cu is introduced.

According to another aspect of the present invention, there is disclosed a pharmaceutical acceptable pharmaceutical composition for diagnosing liver cirrhosis, which comprises a probe for detecting hydrogen sulfide comprising a complex in which the radioisotope Cu is introduced.

According to another aspect of the present invention, there is provided a pharmaceutical acceptable asthma diagnostic pharmaceutical composition comprising a probe for detecting hydrogen sulfide comprising a complex in which the radioisotope Cu is introduced.

According to another aspect of the present invention, there is provided a pharmaceutical composition for diagnosing Parkinson's disease, which comprises a probe for detecting hydrogen sulfide comprising a complex in which the radioisotope Cu is introduced.

According to another aspect of the present invention, there is provided a probe for detecting a hydrogen sulfide comprising a complex in which a radioisotope Cu is introduced, or a pharmaceutically acceptable salt thereof, in an amount of 1 ng to 100 mg, A kit for manufacturing a radioactive isotope Cu labeled medicament in the form of a non-heat-sterilized sterilized state is disclosed.

In one aspect of the present invention, the pharmaceutical preparation kit further comprises 0.01 mL-10 mL of a buffer solution having a pH of 1-9 and a concentration of 1 μM-10 μM.

In another aspect of the present invention, the buffer solution is selected from the group consisting of acetic acid, phosphoric acid, citric acid, fumaric acid, butyric acid, succinic acid, tartaric acid, carbonic acid, glucoheptonic acid, gluconic acid, glucuronic acid, Salt or potassium salt.

In another aspect of the present invention, the concentration of the hydrogen sulfide in the blood or tissue is measured using the complex with the radioisotope Cu introduced thereto, thereby detecting whether the hydrogen sulfide concentration is sharply increased or decreased as compared with the steady state A method for early diagnosis of disease in an animal other than a human is disclosed.

Hereinafter, the present invention will be described in more detail with reference to Examples and the like, but the scope and content of the present invention can not be construed to be limited or limited by the following Examples. It will be apparent to those skilled in the art that various modifications and variations can be made in the present invention without departing from the spirit and scope of the present invention as set forth in the following claims. It is natural that it belongs to the claims.

Synthesis of various chelates

(2R, 3S, 4S, 5R , 6R) -2- ( hydroxymethyl) -6- (1,4,7,10- tetraaza bicyclo dodecane-1-yl) tetrahydro -2 H - pyran -3 , 4,5-triol (CyclenGlc)

Cyclen (240 mg, 1.4 mmol) was added to stir the glucose solution (50 mg, 0.28 mmol) in anhydrous MeOH (10 mL) and the resultant was refluxed under N ₂ for 16 h Respectively. It was confirmed that the reaction starting compound was completely consumed through TLC [stationary phase = C-18 TLC, mobile phase = MeOH: 10% NH ₄ OAc (1: 1)]. The solvent was then evaporated to dryness under reduced pressure conditions and then purified using column chromatography (52 mg, 56%). _{^{1 H NMR (D 2 O,}} 400 MHz): δ 1.8 (s, 3H), 2.60-3.02 (m, 17H), 3.24-3.42 (m, 6H), 3.58-3.63 (m, 1H), 3.78 (d , J = 12.0 Hz, 1 H), 4.00 (d, J = 8.4 Hz, 1 H); ¹³ C NMR (D2O, 100 MHz):? 43.67, 44.97, 45.92, 46.85, 49.18, 61.08, 69.96, 70.29, 77.47, 91.69; HRMS (FAB): m / z calcd for C 14 H 30 N 4 O 5 [M + H] 335.2294; found 335.2296.

Ethyl 3- (1,4,7,10- Tetraazacyclododecane -1 day) Propanoate  synthesis

K ₂ CO ₃ (300 mg, 2.2 mmol) and ethyl 3-bromopropanoate (100 mg, 2.2 mmol) were added to a stirred solution of cyclohexane solution (380 mg, 2.2 mmol) in anhydrous CH ₃ CN (10 mL) 0.61 mmol), and the resultant was stirred at room temperature for 16 hours under N ₂ condition. It was confirmed that the starting compound was completely consumed through TLC (fixed phase = basic alumina TLC, mobile phase = CH ₂ Cl ₂ : MeOH (10: 1)). The reaction was separated using a filter from K ₂ CO ₃ , evaporated to dryness under reduced pressure conditions, and purified by column chromatography (100 mg, 64%). ¹ H NMR (CDCl ₃ , 400 MHz):? 1.27 (t, J = 7.2 Hz, 3H), 2.38-2.57 (m, 10H), 2.64-2.67 (m, 4H), 2.76-2.84 ), 4.07-4. 18 (m, 2H); ¹³ C NMR (CDCl3, 100 MHz):? 14.10, 32.55, 44.38, 45.64, 46.39, 49.61, 50.92, 60.31, 172.67.

Synthesis of 1,4,7,10 -tetrakis (anthracene-9- ylmethyl ) -1,4,7,10 -tetraazacyclododecane

To give to a stirred solution between clan (100 mg, 0.58 mmol) in anhydrous toluene _{(15 mL), K 2 CO} 3 (400 mg, 2.9 mmol) with 9- (chloromethyl) anthracene (655 mg, 2.9 mmol) And the resultant was refluxed under N ₂ condition for 16 hours, and heat was applied. TLC (stationary phase = Silica TLC, mobile phase = EtOAc). The reaction was evaporated under reduced pressure and dried, and then dichloromethane (50 mL) was added. The reaction was stirred well to separate water and organic solvent layers and dried using MgSO ₄ . Pure compounds were then obtained using column chromatography (270 mg, 50%). ¹ H NMR (CDCl ₃ , 400 MHz):? 2.42 (s, 16H), 4.45 (s, 8H), 7.43-7.65 (m, 16H), 7.81-8.09 (m, 10 H); ¹³ C NMR (CDCl ₃ , 100 MHz):? 45.28, 51.02, 54.68, 123.86, 124.60, 125.15, 126.52, 126.82, 127.38, 128.44, 129.31, 129.51, 131.44, 131.49, 131.51, 134.28.

1- (Anthracene-9- Yl methyl ) -1,4,7,10- Tetraazacyclododecane  synthesis

Chloromethylanthracene (1.0 eq.) Was dissolved in toluene (5 mL), acyclen (5.0 eq.) Was added thereto, and the reaction was refluxed for 8 hours to heat. _{^{1 H NMR (CDCl 3, 400}} MHz): δ2.04 (s, 3H, NH), 2.44-2.47 (m, 8H), 2.84-2.94 (m, 8H), 4.75 (s, 2H), 7.46-7.59 (m, 4H), 8.01-8.03 (m, 2H), 8.45-8.51 (m, 3H).

Synthesis of 5 - ((1,4,7,10 -tetraazacyclododecan- 1-yl) sulfonyl ) -N, N -dimethylnaphthalen- 1 -amine

Dissolve chloride (1.0 eq) was dissolved in toluene (5 mL), then cycline (5.0 eq) was added, and the reaction was stirred at room temperature for 2 hours. The precipitated salts were removed and the organic phase separated and dried. The solid material was purified using column chromatography. _{^{1 H NMR (CDCl 3, 400}} MHz): δ2.00-2.04 (m, 3H), 2.89 (s, 6H), 3.11-3.88 (m, 16H), 7.20 (d, J = 7.6 Hz, 1H), 7.50-7.59 (m, 2H), 7.92 (d, J = 7.2 Hz, 1H), 8.34 (d, J = 8.4 Hz, 1H), 8.57 (d, J = 8.8 Hz, 1H); _{^{13 C NMR (CDCl 3, 100}} MHz): δ45.47, 47.37, 49.95, 50.74, 51.90, 115.67, 118.90, 123.12, 127.97, 128.75, 130.28, 130.56, 131.03, 132.84, 152.07.

1,4,7- Trutia -10- Azacyclododecane  synthesis

Step I: slowly add 3-thia-1,5-pentanedithiol (a, 1 eq) and N-boc-bis (2-chloroethyl) amine (b, 1 eq) dissolved in DMF and add Cs ₂ CO ₃ (1.5 eq.). N-Boc- [12] aneNS3 (c) was prepared by reacting at 65 ° C for 96 hours. The solvent was removed in vacuo and the reaction was dissolved in CH ₂ Cl ₂ and then washed with water to remove the cesium salt. After drying with MgSO ₄ , the mixture was filtered and evaporated. The reaction product was recrystallized from warm toluene and purified to obtain Nboc- [12] aneNS3 (c) white crystals.

Step II: N-Boc- [12] aneNS3 (c) was deprotected at room temperature for 15 min with a mixture of TFA and CH ₂ Cl ₂ (1: 1). Excess TFA and CH ₂ Cl ₂ were evaporated, then water was added and the pH of the reaction was adjusted to 14 using Na ₂ CO ₃ and NaOH. After extraction with CH ₂ Cl ₂ , the mixture was dried using MgSO ₄ , then filtered and evaporated to obtain [12] aneNS 3 white crystals. ^{1 H NMR (CDCl3, 400 MHz} ): δ2.71-2.91 (m, 16H), 3.98 (brs, 1H); ≪ ¹³ > C NMR (CDCl3, 100 MHz): [delta] 49.75, 32.37, 31.83, 31.56.

(1Z, N'E) -N '- ((E) -3 - (( bis (dimethylamino) methylene) hydroxy rajo furnace) butane-2-ylidene) Synthesis of -N- methyl-carbamoyl hydroxy acid phenothiazine rajo

Step I: First, semicarbazide (1.0 eq.) And 2,3-butadione (10 eq.) Were mixed in MeOH (10 mL) and the reaction was refluxed for 1 h to yield compound c. It was then purified using column chromatography.

Step II: Compound c (1 eq.) Was first mixed in hydrazine hydrate (10 eq.) And MeOH (10 mL), and the reaction was refluxed for 1 hour to obtain compound c. It was then purified using column chromatography.

Step III: Compound d (1 eq.) Was first mixed in N, N-dimethylurea (1 eq.) And MeOH (10 mL) and the reaction was refluxed for 1 h to give compound f. The solvent was then dried and purified using column chromatography to give pure compound f. ¹ H NMR (DMSO-d 6, 400 MHz):? 1.574 (s, 1H), 1.85 (s, (s, 3H), 3.04 (s, 3H), 3.13 (s, IH).

One- methyl -1,4,7- Triazonan  synthesis

Potassium carbonate (27 mg, 0.19 mmol) and methyl iodide (28 mg, 0.19 mmol) were added to stir TACN (100 mg, 0.78 mmol) in anhydrous acetonitrile (10 mL) The resultant was stirred at room temperature for 16 hours. It was confirmed that a new reaction product was formed through TLC [stationary phase = Silica TLC, mobile phase = dichloromethane: IPA (10: 1)]. The reaction was diluted with acetonitrile, separated from potassium carbonate using a filter, evaporated under reduced pressure conditions and dried to obtain 1-methyl-1,4,7-triazonan. HR-MS (FAB) m / z calc. C ₇ H ₁₇ N ₃ [M + H] ⁺ 144.1501, found 144.1501.

1- (Anthracene-9- Yl methyl ) -1,4,7- Triazonan  synthesis

Potassium carbonate (27 mg, 0.19 mmol) and 9- (chloromethyl) anthracene (43 mg, 0.19 mmol) were added to a stirred solution of TACN (100 mg, 0.78 mmol) in anhydrous acetonitrile After completion of the reaction, the resultant was stirred at room temperature for 16 hours. It was confirmed that a new reaction product was formed through TLC [stationary phase = Silica TLC, mobile phase = dichloromethane: IPA (10: 1)]. The reaction was diluted with acetonitrile, separated from potassium carbonate using a filter, and evaporated under reduced pressure to dryness to give 1- (anthracene-9-ylmethyl) -1,4,7-triazonan . HR-MS (FAB) m / z calc. C ₂₁ H ₂₅ N ₃ [M + H] ⁺ 320.2127, found 320.2127.

One- Hexadecyl -1,4,7- Triazonan  synthesis

To stir TACN (100 mg, 0.78 mmol) in anhydrous acetonitrile (10 mL), potassium carbonate (27 mg, 0.19 mmol) and 1-bromohexadecane (58 mg, 0.19 mmol) were added thereto, and the resultant was stirred at room temperature for 16 hours. It was confirmed that a new reaction product was formed through TLC [stationary phase = Silica TLC, mobile phase = dichloromethane: IPA (10: 1)]. The reaction was diluted with acetonitrile, separated from potassium carbonate using a filter, evaporated under reduced pressure conditions and dried to obtain 1-hexadecyl-1,4,7-triazonan. HR-MS (FAB) m / z calc C ₂₂ H ₄₇ N ₃ [M + H] ⁺ 354.3848, found 354.3848.

1,4,7- Trimethyl -1,4,7- Triazonan  synthesis

To stir TACN (100 mg, 0.78 mmol) in anhydrous acetonitrile (10 mL), potassium carbonate (433 mg, 3.12 mmol) and methyl iodide (443 mg, 3.12 mmol) were added, and the resultant was stirred for 16 hours. It was confirmed that a new reaction product was formed through TLC [stationary phase = Silica TLC, mobile phase = dichloromethane: IPA (10: 1)]. The reaction was diluted with acetonitrile, separated from potassium carbonate using a filter, evaporated under reduced pressure conditions and dried to obtain 1,4,7-trimethyl-1,4,7-triazonan . MS (FAB) m / z calc. C ₉ H ₂₁ N ₃ [M] ⁺ 171.17, measured 171.11

1,4,7- Tris (Anthracene-9- Yl methyl ) -1,4,7- Triazonan  synthesis

Potassium carbonate (433 mg, 3.12 mmol) and 9- (chloromethyl) anthracene (705 mg, 3.12 mmol) were added to a stirred solution of TACN (100 mg, 0.78 mmol) in anhydrous acetonitrile After the addition, the resultant was stirred for 16 hours. It was confirmed that a new reaction product was formed through TLC [stationary phase = Silica TLC, mobile phase = dichloromethane: IPA (10: 1)]. The reaction was diluted with acetonitrile, separated from potassium carbonate using a filter, and evaporated under reduced pressure to dryness to give 1,4,7-tris (anthracen-9-ylmethyl) -1,4- 7-triazonane could be obtained. HR-MS (EI) m / z calc. C ₅₁ H ₄₅ N ₃ [M] ⁺ 699.3613, measured 699.3612.

1,4,7- Trihexadecyl -1,4,7- Triazonan  synthesis

To a stirred solution of TACN (100 mg, 0.78 mmol) in anhydrous acetonitrile (10 mL) was added potassium carbonate (433 mg, 3.12 mmol) and 1-bromohexadecane (949 mg, 3.12 mmol) After that, the resultant was stirred for 16 hours. It was confirmed that a new reaction product was formed through TLC [stationary phase = Silica TLC, mobile phase = dichloromethane: IPA (10: 1)]. The reaction was diluted with acetonitrile, separated from potassium carbonate using a filter, evaporated under reduced pressure conditions and dried to give 1,4,7-trihexadecyl-1,4,7-triazonan I could. HR-MS (EI) m / z calculated value C ₅₄ H ₁₁₁ N ₃ [M + H] < ⁺ >, 802.8856, measured value 802.8856.

⁶⁴ Cu  Radiolabels for various chelates used

Add ⁶⁴ μCiCl ₂ of 0.01 N HCl (1-2 L, 1-5 mCi) to various chelates (100 μg) dissolved in 100 μL 0.1 M ammonium acetate (pH = 6.8) without addition of carrier and incubate for 20 min at 60 ° C By reacting, various kinds of chelates and ⁶⁴ Cu complexes could be made. (Of the chelates, PCB cyclam, a complex of the following Example 19, was allowed to react for 60 minutes at 90 ° C.) The chelate-retained solution (20 μg / μL) ) Or anhydrous DMSO (mixture of Examples 24, 25, 33, 34, 17, 18 and 18-1, 26, and 23 below). For the formation of the corresponding ⁶⁴ Cu complexes, the radioactivity-TLC was used to confirm. For the mixture of Example 17 and 18-1, which is a mixture, the radiolabeled compound was purified using a C-18 Sep Pak column (washed with 10 mL of water to remove DMSO and the compound was eluted with EtOH 200 I eluted six times in μL.)

Example One: ⁶⁴ Cu  Complex-1a

0.01, 2 mg of 1, 4, 7, 10-tetraazacyclododecane was dissolved in 0.1 M NH ₄ OAc (pH 6.8) buffer solution, and then ⁶⁴ CuCl ₂ The solution was added with 10 - 10,000 μCi, and the reaction was carried out at 60 ° C. to obtain the ⁶⁴ Cu complex represented by Formula 1a. If the chelate was not well dissolved, a small amount of DMSO was used.

The labeled yield of the ⁶⁴ Cu complex represented by Formula 1a was confirmed by radio-TLC. The results are shown in Table 1 below.

Reg (mm)
Start (mm)
Stop (mm)
Centroid Rf Region counts Region
CPM % of
Total % of
ROI Rgn 1 48.2 73.1 62.2 0.271 167686.0 83843.0 97.67 100.00 1 peaks - - - - 167686.0 83843.0 97.67 100.00

As shown in Table 1, it was confirmed that ⁶⁴ Cu was labeled with 1, 4, 7, 10-tetraazacyclododecane at 100% purity.

Example 2: ⁶⁴ Cu  Complex-1b

The same procedure as in Example 1 was followed to obtain the ⁶⁴ Cu complex compound represented by Formula 1a.

Example 3: ⁶⁴ Cu  Complex-1c

The same procedure as in Example 1 was carried out to obtain ⁶⁴ Cu complex represented by Formula 1c.

Example 4: ⁶⁴ Cu  Complex-1d

The same procedure as in Example 1 was conducted to obtain a ⁶⁴ Cu complex compound represented by Formula 1d.

Example 5: ⁶⁴ Cu  Complex-1e

The same procedure as in Example 1 was carried out to obtain ⁶⁴ Cu complex represented by Formula 1e.

Example 6: ⁶⁴ Cu  Complex-1f

The same procedure as in Example 1 was followed to obtain the ⁶⁴ Cu complex compound represented by Formula 1f.

Example 7: ⁶⁴ Cu  Complex-1g

The same procedure as in Example 1 was carried out to obtain a ⁶⁴ Cu complex compound represented by the formula 1g.

Example 8: ⁶⁴ Cu  Complex-1h

The same procedure as in Example 1 was carried out to obtain ⁶⁴ Cu complex represented by Formula 1h.

Example 9: ⁶⁴ Cu  Complex-1i

The same procedure as in Example 1 was followed to obtain a ⁶⁴ Cu complex represented by Formula 1i.

Example 10: ⁶⁴ Cu  Complex-1j

The same procedure as in Example 1 was followed to obtain the ⁶⁴ Cu complex compound represented by Formula 1j.

Example 11: ⁶⁴ Cu  Complex-1k

The same procedure as in Example 1 was followed to obtain the ⁶⁴ Cu complex compound represented by Formula 1k.

Example 12: ⁶⁴ Cu  Complex-1l

The same procedure as in Example 1 was carried out to obtain ⁶⁴ Cu complex represented by Formula 11.

Example 13: ⁶⁴ Cu  Complex-1m

The same procedure as in Example 1 was followed to obtain a ⁶⁴ Cu complex represented by Formula 1m.

Example 14: ⁶⁴ Cu  Complex-1n

The same procedure as in Example 1 was carried out to obtain ⁶⁴ Cu complex compound represented by Formula 1n.

Example 15: ⁶⁴ Cu  Complex-1o

The same procedure as in Example 1 was conducted to obtain a ⁶⁴ Cu complex represented by Formula 1o.

Example 16: ⁶⁴ Cu  Complex-1p

The same procedure as in Example 1 was followed to obtain the ⁶⁴ Cu complex represented by Formula 1p.

Example 17: ⁶⁴ Cu  Complex-1q

The same procedure as in Example 1 was followed to obtain the ⁶⁴ Cu complex represented by Formula 1q.

Example  18 ⁶⁴ Cu  Complex-1r

The same procedure as in Example 1 was followed to obtain a ⁶⁴ Cu complex represented by Formula 1r.

Example  18- One: ⁶⁴ Cu  The complex-1r '

The same procedure as in Example 1 was carried out to obtain a ⁶⁴ Cu complex represented by the formula 1r '.

Example 19: ⁶⁴ Cu  Complex-1s

The same procedure as in Example 1 was carried out to obtain ⁶⁴ Cu complex represented by Formula 1s.

Example 20: ⁶⁴ Cu  Complex-2a

The same procedure as in Example 1 was carried out to obtain ⁶⁴ Cu complex represented by Formula 2a.

Example 21: ⁶⁴ Cu  Complex-2b

The same procedure as in Example 1 was carried out to obtain ⁶⁴ Cu complex compound represented by Formula 2b.

Example 22: ⁶⁴ Cu  Complex-2c

The same procedure as in Example 1 was followed to obtain the ⁶⁴ Cu complex compound represented by Formula 2c.

Example 23: ⁶⁴ Cu  Complex-1t

The same procedure as in Example 1 was carried out to obtain a ⁶⁴ Cu complex represented by Formula 1t.

Example 24: ⁶⁴  Cu complex-1u

The same procedure as in Example 1 was followed to obtain a ⁶⁴ Cu complex represented by Formula 1u.

Example 25: ⁶⁴ Cu  Complex-4

The same procedure as in Example 1 was carried out to obtain the ⁶⁴ Cu complex compound represented by Formula 4 above.

Example 26: ⁶⁴ Cu  Complex-1 w

The same procedure as in Example 1 was carried out to obtain ⁶⁴ Cu complex represented by Formula 1w.

Example 27: ⁶⁴ Cu  Complex-2d

The same procedure as in Example 1 was followed to obtain the ⁶⁴ Cu complex compound represented by Formula 2d.

Example 28: ⁶⁴ Cu  Complex-2e

The same procedure as in Example 1 was conducted to obtain ⁶⁴ Cu complex compound represented by Formula 2e.

Example 29: ⁶⁴ Cu  Complex-2f

The same procedure as in Example 1 was followed to obtain the ⁶⁴ Cu complex compound represented by Formula 2f.

Example 30:  ⁶⁴ Cu  Complex-2g

The same procedure as in Example 1 was carried out to obtain a ^{6 4} Cu complex represented by the formula 2g.

Example 31: ⁶⁴ Cu  Complex-2h

The same procedure as in Example 1 was conducted to obtain ⁶⁴ Cu complex represented by Formula 2h.

Example 32: ⁶⁴ Cu Complex -2i

The same procedure as in Example 1 was carried out to obtain ⁶⁴ Cu complex represented by Formula 2i.

Example 33: ⁶⁴ Cu  Complex-3a

The same procedure as in Example 1 was carried out to obtain ⁶⁴ Cu complex represented by Formula 3a.

Example 34: ⁶⁴ C  u complex compound-3b

The same procedure as in Example 1 was followed to obtain the ⁶⁴ Cu complex compound represented by Formula 3b.

Experimental Example  1: Sensitivity test for detection of hydrogen sulfide

The following experiments were conducted to determine the sensitivity of detecting the hydrogen sulfide of ⁶⁴ Cu-labeled complexes according to the present invention. The results are shown in Tables 2 and 3 and FIG.

Approximately 45-55 Ci of the ⁶⁴ Cu-labeled complex (250 μL tertiary distilled water) was mixed with various concentrations of NaHS solution (250 L tertiary distilled water) and then stirred at 37 ° C for 15 minutes After the reaction was completed, the reaction was stirred vigorously to ensure proper mixing.

The formation of copper sulfide ( ⁶⁴ CuS) was quantified by using a radio-TLC scanner and the results are shown in Tables 2 and 3 below.

Example 1 Example 2 Example 3 Example 23 Example 16 Example 20 Example 21 NaHS
(μM) cyclen cyclen-Anthracene
(1: 1) DODACD ATTCD cyclam TACN TACD 0 0 0 0 0 0 0 0 One 3 0 0 0 0 0 0 10 11 0 3 0 0 9 6.2 15 63 0 6 36.4 0 11 10 20 95 15 79 49.4 0 13.5 58 50 100 56 100 67.7 1.3 24.6 63 100 100 100 100 78.2 10.5 54.4 100

Example % De-complexation with 100 μM NaHS solution 24 0 25 0 33 26.6 34 0 17 39.67 26 62.43 + - 0.87 23 78.3 ± 2.6

As shown in Tables 2 and 3, the lowest concentration of hydrogen sulfide detected using the ⁶⁴ Cu-labeled complex according to the present invention was measured. As a result, the ⁶⁴ Cu-labeled cyclen of Example 1 according to the present invention , The detection limit from low concentration to high concentration was the lowest, and it was confirmed that sensitivity to hydrogen sulfide was excellent.

In addition, as shown in Figures 2a and 2b, another embodiment according to the present invention in other sulfide ion concentration in Example 1, ⁶⁴ Cu- confirming a radio-TLC results measured using a labeled result of cyclen, ⁶⁴ of the first embodiment The Cu-labeled cyclen exhibits Rf values greater than 0, but when ⁶⁴ CuS is formed by reaction with sulfide ions, the Rf value is changed to 0, which is located at the origin without moving in the radio-TLC.

Experimental Example  2: Detection of hydrogen sulfide by time

The following experiments were carried out to investigate the reactivity of the ⁶⁴ Cu-labeled complexes according to the invention over time. The results are shown in Table 4 and FIG.

The detailed experimental method is as follows.

Approximately 45-50 μCi of a ⁶⁴ Cu-labeled complex (250 μL of tertiary distilled water) was mixed with 100 μM NaHS solution (250 mL tertiary distilled water), followed by stirring at 37 ° C for 15 minutes, ⁶⁴ CuS) was quantitated using a radio-TLC scanner.

division 10 seconds 30 seconds 60 seconds 5 minutes 10 minutes 15 minutes Example 1 cyclen 13 17 21 100 100 100 Example 2 cyclen-Anthracene 21 22 33 82 100 100 Example 3 DODACD 62 70 75 100 100 100 Example 16 cyclam 0 0 0 3 5 10 Example 20 TACN 0 0 5 13 60 60 Example 21 TACD 52 72 100 100 100 100

As shown in Table 4, in order to confirm the reaction rate of the ⁶⁴ Cu-labeled complex compound according to the present invention with the sulfide ion, the concentration of hydrogen sulfide over time was measured. As a result, ⁶⁴ In the case of the Cu-labeled complex, it was confirmed that the detection of hydrogen sulfide started within about 10 seconds.

Experimental Example  3: Selectivity test for hydrogen sulfide

Since there are many other anions and radicals besides hydrogen sulfide in the body, the following experiment was conducted to confirm the selective reactivity of ⁶⁴ Cu-labeled complexes according to the present invention with sulfide ions. The results are shown in Table 5 below.

Approximately 45-55 μCi of ⁶⁴ Cu-labeled complex (250 μL of tertiary distilled water) was mixed with various anion solutions (250 μL of tertiary distilled water) containing biotiol at a concentration of 100 μM, Lt; / RTI >

The formation of copper sulfide ( ⁶⁴ CuS) was quantified using a radio-TLC scanner.

Various biologic thiol or salt solutions were prepared by dissolving the required compounds in tertiary distilled water.

* In the table above, the concentration of all biothreitives and other competing substances is 100 μM. Specific concentrations of biothiol, anion, and oxidant used in the above table are as follows. [L-Cys (1 mM); L-Hcys (10 mM); GSH (10 mM); DL-dithiothreitol (DTT, 100 [mu] M); L-ascorbic acid (L-AsA, 10 mM); 2-mercaptoethanol (2-ME, 10 mM); NaCl (10 mM); KI (1 mM); Na ₂ S ₂ O ₃ (1 mM); Na ₂ S ₂ O ₅ (1 mM); NaOAc (1 mM); H ₂ O ₂ (100 M); NaClO ₄ (100 μM); NaHCO ₃ (100 μM); NaNO ₂ (100 M); NaN ₃ (100 [mu] M); ATB 337 (100 [mu] M); diallyl trisulphide (100 [mu] M)].

As shown in Table 5 above, the degree of reaction with various anions containing sulfur was measured. As a result, the ⁶⁴ Cu-labeled complexes of Examples 1, 2, 16 and 20 according to the present invention were found to selectively react with hydrogen sulfide On the other hand, in the case of the ⁶⁴ Cu-labeled complexes of Examples 3 and 21, it was confirmed that not only the hydrogen sulfide was selectively reacted but also the other biothiol compounds and anions.

As a result of confirming the selective reactivity with the hydrogen sulfide using the radioactive copper ion not labeled with the chelate, unlike the ⁶⁴ Cu-labeled cyclen of Example 1 according to the present invention, the copper ion has various bio thiol compounds or anions As shown in FIG.

Experimental Example  4: Cell uptake experiment

Since the degree of cell uptake of the drug is an important prognostic factor in diagnosing the disease, in order to determine the degree of ingestion of the ⁶⁴ Cu-labeled complex according to the present invention, the degree of change in the degree of ingestion of the cell under various hydrogen sulfide concentration conditions is measured Experiments were performed.

After plating 5 x 10 ⁶ CT-26 colon cancer cells on each plate, the ⁶⁴ Cu-labeled complexes of Examples 1, 7, and 9 according to the present invention were treated with 10 - 300 μCi, After incubation for a few minutes, the cells were washed with PBS solution, and then added with 0, 1, 50 μM NaHS solution, followed by incubation for 5 minutes. After washing the cells again with PBS, trypsin and EDTA were added to remove the cells from the plate. After centrifugation, the supernatant was removed, and the radioactivity of the remaining cells was measured by a gamma counter.

The results are shown in Table 6 below.

division Concentration (μM) Intake Example 1 0 600 One 780 50 850 Example 7 0 20000 One 21000 50 23000 Example 9 0 7000 One 6500 50 6900 Example 20 0 30000 One 32000 50 34000

As shown in Table 6 above, in the case of the ⁶⁴ Cu-labeled complexes according to the present invention, it was confirmed that as the concentration of NaHS increases, the radioactivity of the ⁶⁴ Cu-labeled complexes consumed in the cells is small but increases.

Experimental Example  4-2: Hydrogen sulfide-specific imaging verification experiment

In order to ensure that the cover ⁶⁴ Cu- complexes according to the present invention can detect hydrogen sulfide optionally in the body, the injection of different materials or the like in the rat, and then the cover ⁶⁴ Cu- complexes according to the invention the tail We performed an experiment to check whether the images were selected through optical imaging.

FIG. 4 shows photographs showing the selective visualization of ⁶⁴ Cu-labeled complexes according to the present invention through optical imaging after injecting the compound of the present invention into a mouse. Referring to FIG. 4, at the start of the first experiment, Matrigel alone, Matrigel + 1 mg NaCl, Matrigel + 0.5 mg NaHS, and Matrigel + 0.05 mg NaHS were injected into each of the four sites of rats, and then the ⁶⁴ Cu-labeled complexes according to Example 1 Was injected with 1.8 mCi of mouse tail, and optical image was measured immediately. As a result, of the four sites, signals were observed only in the lower two sites injected with NaHS which generates hydrogen sulfide. In addition, it was confirmed that the signal was still observed in the lower two sites after 4 hours.

Next, at the start of the second experiment, similarly, four regions of the rats were treated with about 0.48 mCi of ⁶⁴ Cu (II) ions, ⁶⁴ CuS, ⁶⁴ Cu-labeled complexes according to Example 1, 16 is a bottom left image of FIG. 4, and after 4 hours has passed, the bottom right image is a photograph taken after the injection of the ⁶⁴ Cu-labeled complex according to FIG. Referring to Figure 4, ⁶⁴ CuS than the complex Cu- cover ⁶⁴ according to the different embodiments and the embodiment ⁶⁴ Cu- labeling complex according to example 1 16 It can be seen that the in vitro release at a very high speed. ⁶⁴ Cu (II) ions showed about half of the residence time of ⁶⁴ CuS. The above experiments show that ⁶⁴ Cu-labeled complexes according to the present invention bind to hydrogen sulfide to produce ⁶⁴ CuS, so that they remain in the body for a long time and can be easily detected through imaging.

Manufacturing example  1: Manufacture of animal models

(1) Model of paw pain inflammation

Inflammatory models were established by injecting small amounts of complete Freund's adjuvant (CFA) or formalin only in one of the paws of the backbone of Balb / c or ICR mice. And were used for imaging or other in vivo experiments 1 day after injection of inflammatory substances.

It is already well known that the concentration of hydrogen sulfide in the inflammation site is high.

(2) myocardial infarction model

Myocardial infarction model was established using Sprague-Dawley rats (over 12 weeks, approximately 250 g). Two days after inducing myocardial infarction by blocking part of the coronary artery in the cardiovascular system, it was used in the imaging experiment. It is known that the concentration of hydrogen sulfide in the myocardial infarction area is high.

(3) Arthritis model

Bovine type II collagen, an arthritis inducer, was intradermally injected twice into the tail of DBA / 1J rats and used for imaging experiments 5 weeks later (approximately 13 weeks old). The degree of arthritis of the foot was quantified by scoring.

(4) brain inflammation model

Sprague-Dawley rats were injected with lipopolysaccharide, an inflammation-inducing substance, on one side of the brain to induce inflammation on one side of the brain. After 24 hours of inflammation induction, they were used for imaging experiments.

(5) tumor model

In order to prepare the CT26 tumor model, the number of cells was calculated using a hemocyte calculator, and 5 × 10 ⁶ cells were subcutaneously injected into the right flank of the BALB / c mouse. After about 10 days, 1 cm of tumor was generated And then used in the experiment.

The EMT6 tumor model was also used to calculate the number of cells using a hemocytometer. 2 × 10 ⁵ cells were subcutaneously injected into the right flank of the BALB / c female mouse and the left shoulder area. After 2 to 3 weeks, When it emerged, it was used for later experiments.

(6) Sepsis model

After the lower abdomen of ICR mice was opened, a small puncture was made using a syringe needle in the appendix and an acute sepsis model was prepared by suturing.

(7) brain tumor model

The surface of the rat's head was incised and the thread vein and the outer membrane of the skull were removed. Subsequently, precise coordinates were determined by stereotaxic surgery and punctures were made. 20,000 C6 cells, known as glial cells of the rat brain, were injected using a syringe pump. After the injection of the cells, the syringe was slowly removed for 5 minutes with the absorption time, the hole was closed with the cemented cement and the suture was made to create a brain tumor model.

(8) Pancreatitis model

Babl / c mice were injected with caerulein (50 μg / kg) seven times at 1 hour intervals by intraperitoneal injection to produce a pancreatitis model.

Experimental Example  5: Optical and Nuclear Medicine imaging studies

Nuclear medical images were measured using Siemens Healthcare's Inveon PET / CT system and optical images were measured using IVIS spectrum CT equipment. Cherenkov light from ⁶⁴ Cu was measured using a high-sensitivity charge-coupled device (CCD) camera.

The imaging time is 1-5 minutes. The amount of radioactivity injected is approximately 50-2000 μCi. The imaging time can be varied as needed, such as immediately after the first injection, 1 hour, 4 hours, 24 hours after the first injection Respectively.

(1) Foot inflammation model

As shown in FIG. 5A, when treated with ⁶⁴ Cu-labeled cyclen of Example 1 according to the present invention, it was confirmed that the right foot, which is an inflammation site, in the PET image showed higher uptake than the other foot, As shown in Fig. 5C, the intake of the ⁶⁴ Cu-labeled cyclen of Example 1 according to the present invention was increased in the inflammatory site induced by CFA in the optical image. As shown in Fig. 5C, Of ⁶⁴ Cu-labeled cyclen was found to be fast enough to confirm the site of inflammation immediately after injection (1.5-10 min).

As shown in Fig. 5D, the same results were also confirmed in the foot inflammation model using ICR mice and Balb / c mice, and the ⁶⁴ Cu-labeled cyclen of Example 1 according to the present invention It has been confirmed that the time remaining in the body of the animal model is more than 24 hours, and the therapeutic effect can be predicted noninvasively without continuously injecting the drug.

In addition, as shown in Figs. 6A to 6C, in the case of the ⁶⁴ Cu-labeled TACN of Example 20, it was confirmed that after one hour, the ingestion of the inflamed foot was higher than that of the other side, The same result was confirmed, and the inflammation site could be confirmed in the image immediately after the ⁶⁴ Cu-labeled TACN injection of Example 20.

In addition, in the case of the ⁶⁴ Cu-labeled complexes of Examples 2, 3, 14, and 21 according to the present invention, as shown in Figs. 7A to 7D and Figs. 8A to 8E, Selective intake was confirmed.

On the other hand, as shown in Figs. 9a and 9b and also in Figs. 10a and 10b, in the case of 1g and 1i of ⁶⁴ Cu-labeled compounds of Examples 7 and 9, the inflammation-induced It was confirmed that the ingestion was higher than the other side of the foot.

On the other hand, as shown in Figs. 11A to 11D, it was confirmed that the ⁶⁴ Cu-labeled cyclam of Example 16 showed no significant difference in the inflammation site and the foot of the normal site, and the same amount of ⁶⁴ Cu- It was confirmed that the low intake was also observed in comparison with the case of injected cyclen.

(2) Arthritis model

When the arthritis model was injected into the ⁶⁴ Cu-labeled cyclen of Example 1 according to the present invention, selective ingestion at the joint site was confirmed, as shown in Figures 13a and 13b.

(3) brain inflammation model

When lPS was injected into the brain half of the mouse to induce inflammation and after 1 day, PET image was taken after the ⁶⁴ Cu-labeled complex compound according to Example 17 according to the present invention was injected, as shown in Fig. 14 Likewise, it was confirmed that the inflammation site showed a higher intake than the other side.

Experimental Example  6: Imaging studies in myocardial infarction model

It is known that the concentration of hydrogen sulfide in the myocardial infarction area is high. The mice that injected ⁶⁴ Cu-labeled complex into the myocardial infarction-occluded macrophages by blocking the coronary arteries of the heart of the rats were injected with 50-900 μCi, The extent of ingestion in the infarct area was monitored by PET imaging.

⁶⁴ Cu-labeled complexes were injected. Two days later, [ ¹⁸ F] FDG was injected to confirm the correct induction of myocardial infarction.

36 hours after the establishment of myocardial infarction model, 730 μCi of the radiolabeled compound according to the present invention was injected, and nuclear medicine images were taken. After 2 days, the same mice were injected with 1.06 μCi of FDG FDG-PET images were obtained. The results are shown in Fig. 15 to Fig.

(In the nuclear medicine image, the upper part is the transverse image and the lower image is the coronal image.)

As shown in FIG. 15, when the ⁶⁴ Cu-labeled cyclen of Example 1 according to the present invention was injected, the myocardial infarction area was clearly observed in the myocardial infarction area In contrast, FDG injection showed no FDG uptake in the myocardial infarction area, suggesting that the myocardium is not a donut shape but a partly broken ring in the FDG image.

In addition, as shown in Fig. 16, the ⁶⁴ Cu-labeled TACN compound of Example 20 also clearly observed a myocardial infarction site.

On the other hand, as shown in Fig. 17, in the case of the ⁶⁴ Cu-labeled cyclam of Example 16, no specific uptake could be found in the heart as in the previously identified foot inflammation model and arthritis model.

Experimental Example  7: Imaging studies in brain tumor model

In the prepared brain tumor model rats, ⁶⁴ CI-labeled 1q compound according to Example 17 was administered, and PET images were obtained after 27 hours. A high level of intake was seen in the brain tumor area, and this imaging experiment shows that a brain tumor can be diagnosed by a hydrogen sulfide detection probe.

Experimental Example  8: Imaging studies in various tumor models

PET images were obtained by injecting the ⁶⁴ Cu-labeled complexes of Example 1 into model rats having the EMT6 tumors and model mice having CT26 tumors. The results are shown in Figs. 19a and 19b. Referring to FIGS. 19A and 19B, it can be confirmed that the tumor area (red circle) shows a higher intake than the background. This demonstrates that hydrogen sulfide detection probes can be used to diagnose various tumors. In particular, EMT6 tumors are widely used as a hypoxic tumor model and thus can be used as probes for detecting hypoxia in tumors.

Experimental Example  9: Imaging studies in pancreatitis model

The model mice having the pancreatitis prepared above were injected with ⁶⁴ Cu-labeled complexes according to Example 1 and Example 20 according to the present invention, and then the optical images and tissues were excised and the obtained PET images were obtained. The results are shown in Fig. Referring to FIG. 20, it can be seen that the ⁶⁴ Cu-labeled complexes according to Examples 1 and 20 show a high uptake in the pancreas (red ellipse) in the pancreatitis model and thus the pancreatitis can be diagnosed through the probe of the present invention .

Experimental Example  10: Imaging studies in sepsis models

The model mice having the prepared sepsis were injected with ⁶⁴ Cu-labeled complexes according to Example 20 according to the present invention to obtain PET images, and the results are shown in FIG. Referring to FIG. 21, it can be seen that the intake of the whole body is increased in the case of the sepsis model. In particular, it can be seen that the ingestion of the lung part is increased, and thus it can be utilized for the diagnosis of sepsis through the hydrogen sulfide probe.

Experimental Example  11: In normal rats  Hydrogen sulfide imaging study

The normal mice were injected with the ⁶⁴ Cu-labeled complex according to Example 1 according to the present invention, and the PET images were obtained. After the tongue was excised, the PET images were obtained again. The results are shown in FIG. PET images before the removal of the tongue showed a high intake of (a) the tongue area, but (b) after the resection showed a decrease in the intake at that site. Also, high intake is seen in the removed tongue. (c) is a photograph image taken before the image is taken by cutting out the tongue. This demonstrates that hydrogen sulfide probes can be used for the detection of hydrogen sulfide in both normal and diseased conditions.

Experimental Example  12: Determination of hydrogen sulfide calibration  Research

FIG. 23 shows a calibration curve obtained by measuring the degree of desorption of the ⁶⁴ Cu-labeled TACN according to Example 20 according to the present invention by reacting with a different concentration of sodium hydrogen sulfide (NaHS). It shows that the concentration of hydrogen sulfide can be measured accurately by measuring the degree of desorption.

Experimental Example  13: Comparison with conventional conventional measurement method of hydrogen sulfide concentration

In order to show that the complex for detecting hydrogen sulfide according to the present invention enables specific detection for models with a specific disease with a higher concentration of hydrogen sulfide than the normal mice, the ⁶⁴ Cu-labeled complex compound according to Example 20 , And data obtained by a conventional conventional methylene blue method were compared.

The obtained data are shown in Table 7 and Fig.

Normal rat
Standard Deviation
Myocardial infarction model
Standard Deviation ⁶⁴ Cu-TACN
23.6
3.9
41.8
2.0
Methylene blue
23.8
7.0
40.3
2.0

Referring to Table 7 and FIG. 24, it can be seen that the complex for detecting hydrogen sulfide according to the present invention can specifically detect only the myocardial infarction model generating hydrogen sulfide, which is comparable to the conventional methylene blue method have.

Claims (30)

A probe for detecting hydrogen sulfide (H ₂ S) comprising a complex in which a radioisotope Cu represented by the following formula (3) is introduced:
(3)

(In the above formula (3), X is as described herein.)
The compound according to claim 2, wherein the compound represented by formula (3)

And

And a probe for detecting hydrogen sulfide.
The method according to claim 1,
A probe for detecting hydrogen sulfide, comprising 50 Ci-1000 μCi / kg of the complex compound of Formula 2 immediately before use.
2. The method of claim 1, wherein the complex compound of Formula 2 is selected from the group consisting of positron emission tomography (PET) contrast agent, gamma camera, SPECT (single photon emission computed tomography), Cherenkov optical image contrast agent, a contrast agent for magnetic resonance imaging, a computed tomography (CT), and a US (Ultrasound).
A method for detecting or imaging sulfide ions in a cell or tissue using a probe for detecting hydrogen sulfide comprising a complex having a radioisotope Cu introduced therein according to claim 1.
6. The method of claim 5,
Wherein the imaging of the sulfide ion is performed by measuring Cherenkov light from the radioactive isotope Cu.
The method according to claim 6,
Wherein the Cherenkov light has a wavelength in the range of 200 to 1000 nm.
6. The method of claim 5,
A method for imaging sulfide ions in a cell or an extracellular matrix, wherein the probe for detecting a hydrogen sulfide comprising a complex in which the radioisotope Cu is introduced locally locates at positions or positions abnormally increased in hydrogen sulfide after administration.
Providing a probe for detecting hydrogen sulfide in a pharmacological carrier comprising a complex having a radioisotope Cu introduced therein according to claim 1; Injecting the probe into a mammal other than a human; And scanning a mammal other than a human using a radiological diagnostic imaging instrument.
A pharmaceutical composition for diagnosing a pharmaceutically acceptable inflammatory disease, comprising a probe for detecting hydrogen sulfide comprising a complex in which the radioisotope Cu of claim 1 is introduced.
11. The method of claim 10, wherein the inflammatory disease is selected from the group consisting of rheumatoid arthritis, non-rheumatoid inflammatory arthritis, arthritis associated with Lyme disease, inflammatory osteoarthritis, meningitis, osteomyelitis, inflammatory bowel disease, appendicitis, pancreatitis, , Inflammatory diseases due to bacterial infection, and the like.
A pharmaceutical composition for diagnosing a pharmaceutically acceptable heart disease comprising a probe for detecting hydrogen sulfide comprising a complex having a radioisotope Cu introduced therein according to claim 1.
The pharmaceutical composition according to claim 12, wherein the heart disease is at least one selected from myocardial infarction, cardiac ischemia, angina pectoris, cardiomyopathy, and endocarditis.
A pharmaceutical composition for the diagnosis of Parkinson's disease comprising a probe for detecting hydrogen sulfide comprising a complex having a radioisotope Cu introduced therein according to claim 1.
A pharmaceutical composition for diagnosing a pharmaceutically acceptable Alzheimer's disease comprising a probe for detecting hydrogen sulfide comprising a complex having a radioisotope Cu introduced therein according to claim 1.
A pharmaceutical composition for the diagnosis of Down's Syndrome comprising a probe for detecting hydrogen sulfide comprising a complex having a radioisotope Cu introduced therein according to claim 1.
A pharmaceutical acceptable diagnostic pharmaceutical composition comprising a probe for the detection of hydrogen sulfide comprising a complex having a radioisotope Cu introduced therein according to claim 1.
18. The pharmaceutical composition according to claim 17, wherein said tumor diagnosis is hypoxia in tumor diagnosis.
A pharmaceutical composition for diagnosing acceptable sepsis, comprising a probe for detecting hydrogen sulfide comprising a complex in which the radioisotope Cu of claim 1 is introduced.
A pharmaceutical composition for the diagnosis of pain comprising a probe for the detection of hydrogen sulfide comprising a complex in which the radioisotope Cu of claim 1 is introduced.
A pharmaceutical composition for the diagnosis of atherosclerosis, which comprises a probe for detecting hydrogen sulfide comprising a complex having a radioisotope Cu introduced therein according to claim 1.
A pharmaceutical acceptable diagnostic pharmaceutical composition for the diagnosis of diabetes comprising a probe for detecting hydrogen sulfide comprising the complex having the radioactive isotope Cu introduced therein according to claim 1.
A pharmaceutical acceptable pharmaceutical composition for the diagnosis of stroke, comprising a probe for detecting hydrogen sulfide comprising a complex having a radioisotope Cu introduced therein according to claim 1.
A pharmaceutical acceptable pharmaceutical composition for the diagnosis of cirrhosis, comprising a probe for detecting hydrogen sulfide comprising a complex having a radioisotope Cu introduced therein according to claim 1.
A pharmaceutical acceptable asthma diagnostic pharmaceutical composition comprising a probe for detecting hydrogen sulfide comprising a complex having a radioisotope Cu introduced therein according to claim 1.
A pharmaceutical composition for the diagnosis of Parkinson's disease comprising a probe for detecting hydrogen sulfide comprising a complex having a radioisotope Cu introduced therein according to claim 1.
A probe for detecting hydrogen sulfide comprising a complex with a radioisotope Cu of claim 1, or a pharmaceutically acceptable salt thereof, in an amount of 1 ng to 100 mg, and which is sealed in a solution state, a freeze state or a freeze- A kit for manufacturing a radioisotope Cu-labeled medicament in a sterilized form.
28. The kit of claim 27, wherein the pharmaceutical preparation kit further comprises 0.01 mL to 10 mL of a buffer solution having a pH of 1-9 and a concentration of 1 μM to 10 M.
29. The method of claim 28, wherein the buffer solution is selected from the group consisting of acetic acid, phosphoric acid, citric acid, fumaric acid, butyric acid, succinic acid, tartaric acid, carbonic acid, glucoheptonic acid, gluconic acid, glucuronic acid, Potassium salt. ≪ RTI ID = 0.0 > 11. < / RTI >
A diagnostic reagent for measuring the concentration of hydrogen sulfide in blood and tissues, comprising a complex in which the radioisotope Cu is introduced according to claim 1.

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