KR20140117199A - Novel benzamide derivative or pharmaceutically acceptable salt thereof, preparation method thereof and pharmaceutical composition for diagnosis of cancer disease containing the same as an active ingredient - Google Patents

Abstract

The present invention relates to a novel benzamide derivative having a macrocycle or a pharmaceutically acceptable salt thereof, a method for producing the same, and a pharmaceutical composition containing the same as an active ingredient for PET diagnosis. The novel benzamide derivative according to the present invention stably forms a complex compound with ^68Ga. The complex compound has a high selective accumulation rate on skin cancer cells, and the complex compound is completely excreted from the body through the kidney, thereby being safe for the human body. Additionally, since the complex compound has a high accumulation rate on skin cancer cells against muscles providing a background for the image and a high accumulation rate on the skin cancer cells according to the elapse of time, the diagnostic efficiency of nuclear medicine imaging is excellent. Therefore, the pharmaceutical composition containing the complex compound can be useful as a pharmaceutical composition for diagnosing skin cancers such as malignant melanoma, basal cell carcinoma, and squamous cell carcinoma.

Description

TECHNICAL FIELD The present invention relates to a novel benzamide derivative or a pharmaceutically acceptable salt thereof, a process for preparing the same, and a pharmaceutical composition for diagnosing skin cancer containing the same as an active ingredient. containing the same as an active ingredient}

The present invention relates to novel benzamide derivative or a pharmaceutically acceptable salt thereof, relates to a production method thereof and a skin cancer diagnostic and therapeutic pharmaceutical composition containing the same as an active ingredient, and more particularly to gallium ⁽⁶⁸ Ga) and to cover A novel benzamide derivative having a structure in which a positive functional chelating agent and a benzamide derivative are linked to a linker thiourea ((NH ₂ ) ₂ C = S), a process for producing the same, and a pharmaceutical composition for diagnosing skin cancer, .

Skin cancer is all skin cancer that occurs in the skin. It is relatively common, but it is not considered as important as gastric cancer, liver cancer, lung cancer, etc. because it is low in the risk of life. It does not require complicated tests or expensive medical equipment compared to other sites of cancer, can be easily found with the naked eye, and can be diagnosed through a simple biopsy. However, in many skin cancer patients, when the skin is first developed, it is difficult to find it early because it is simply a dermatitis, a spot or a boil, so it is difficult to cure the disease and sometimes the life is lost.

There are many different types of skin cancer, depending on what type of skin cells it has. Among these many skin cancer, there is skin cancer that is not easily metastasized and easy to cure. Therefore, diagnosing what kind of skin cancer is very important for the prognosis of skin cancer.

It is generally known that the majority of skin cancer is basal cell carcinoma, squamous cell carcinoma and melanoma. Among these melanomas, melanocytes or melanocytes, which are melanocyte-producing skin cells, are malignant tumors that occur in any organs in which melanocytes are present, but are most prevalent especially in skin. Generally, it is common in Caucasians and rarely in Asians or Blacks, but it is more likely to develop if you work a lot in the field.

The etiology of these melanomas has not been clearly elucidated, but genetic factors and ultraviolet exposure are assumed to be involved. It has been reported that the incidence of black papules in parents and children is 8 times higher than that in non-parents. In addition, 20% -50% of melanomas originate from existing points, particularly from birth to birth, and from unusual points.

Most melanomas can be cured by surgical resection of skin cancer at the time of initial detection, but they are not asymptomatic such as itching or pain, and appear as yellowish brown or black spots or nodules. (3 stages), the survival rate is reduced to less than 50% (Non-Patent Documents 1 to 3). Therefore, melanoma is important in the treatment after onset, but the development of melanoma-specific markers remains one of the main tasks of melanoma research, because it can increase the survival rate of malignant melanoma patients through faster and more accurate diagnosis than anything else.

Positron Emission Temography (PET) imaging, on the other hand, is a tomographic nuclear imaging technique using radioactive tracer molecules that emit positrons, a quantitative imaging method that allows the measurement of the local concentration of the radioactive tracer molecule to be. When a positron encounters an electron, both are extinguished, resulting in the release of energy in the form of a gamma ray, which is detected by a positron emission tomography (PET) scanner. Positron emission tomography (PET) uses natural materials used by the body as tracer molecules to provide information about the physiological function of a particular area of the body or body, as well as providing information about the structure of the body . Typical tracer molecules include, for example, 2-fluoro-2-deoxy-D-glucose (FDG), which is similar to naturally occurring glucose, to which ¹⁸ fluorine ( ¹⁸ F) is added. The choice of the tracer molecule depends on what you are scanning. In general, a tracer that is capable of accumulating in a region of interest or selectively absorbed by a particular type of tissue, e. G., Skin cancer cells, is selected. Scanning consists of a static image or dynamic series obtained after a period of time during which the radioactive tracer molecule enters a biochemical process of interest and the scanner detects the spatial and temporal distribution of the tracer molecule.

Typically, radionuclides used as positron emission tomography (PET) tracers are carbon ( ¹¹ C), fluorine ( ¹⁸ F), oxygen ( ¹⁵ O), nitrogen ( ¹³ N) or bromine ( ⁷⁶ Br) Recently, a new positron emission tomography (PET) tracer based on a radioactive isotope labeled metal complex containing both a functional chelating agent and a radioactive metal has been produced. A bi-functional chelating agent is a chelator that coordinates to a metal ion and is linked to a targeting vector capable of binding to a target site in the body of a patient. Such a targeting vector may be a peptide that binds to a particular receptor in a particular region of the body or that is associated with a particular disease. In addition, the targeting vector may be, for example, an oligonucleotide specific for an activated tumor gene. An advantage of such a complex is that both functional chelating agents can be labeled with a variety of radioactive metals, for example gallium ( ⁶⁸ Ga), bismuth ( ²¹³ Bi) or iridium ( ⁸⁶ Y). In this way, radioisotope labeled complexes with particular properties can be "tailored " for specific applications.

Accordingly, studies on diagnostic markers including both functional chelating agents have been actively conducted, and the results are as follows.

First, a pharmaceutical composition for detecting positron emission tomography (PET) in which a positive functional chelating agent is bound to a human-safe peptide compound and then a radioisotope is labeled with germanium / gallium ( ⁶⁸ Ge / ⁶⁸ Ga) (Patent Document 1).

In addition, diagnostic markers that improve the fluorescence intensity of a marker by binding nanoparticles to a bi-functional chelating agent have been known, and a bi-functional chelating agent or mannosyl is bound to human serum albumin for immunocyte imaging Known diagnostic markers and kits labeled with gallium ( ⁶⁸ Ga) have been known (Patent Documents 2 and 3).

Furthermore, a pharmaceutical composition for the treatment or diagnosis of gallium ( ⁶⁸ Ga) -bifunctional chelating agent-epidermal growth factor receptor (hEFR) developed for diagnosis and treatment of skin cancer has been known (Patent Document 4).

Therefore, the inventors of the present invention have been studying a marker for diagnosing skin cancer containing both functional chelating agents, and found that a novel benzamide derivative containing both functional chelating agents is easily labeled with gallium ( ⁶⁸ Ga) and has excellent stability of the labeled complex And it was confirmed that the rate of accumulation on skin cancer cells was high, and the present invention was completed.

Korean Patent Publication No. 2004-0068363; Korean Patent Publication No. 2010-0030195; Korean Patent Publication No. 2010-0087511; Korea Patent Publication No. 2007-0106731.

Carlson, J. A. et al., J. Am. Acad. Dermatol. 2005, 52, 743; Atallah, E. et al., Curr. Treat. Options Oncol. 2005, 6, 185; Gogas, H. J. et al., Cancer. 2007, 109, 455.

It is an object of the present invention to provide a novel benzamide derivative or a pharmaceutically acceptable salt thereof, to which both functional chelating agents are bonded.

It is another object of the present invention to provide a process for the preparation of the novel benzamide derivatives.

It is another object of the present invention to provide a complex in which the radioisotope is labeled with the novel benzamide derivative.

Another object of the present invention is to provide a pharmaceutical composition for skin cancer diagnosis containing the above-mentioned complex as an active ingredient.

It is still another object of the present invention to provide a skin cancer diagnosis kit comprising the complex as an active ingredient.

In order to achieve the above object,

The present invention provides a novel benzamide derivative or a pharmaceutically acceptable salt thereof, to which both functional chelating agents represented by the following general formula (1) are bonded:

[Chemical Formula 1]

은 본 명세서에서 정의한 바와 같다).
(Wherein R and < RTI ID = 0.0 >

Are as defined herein.

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

There is provided a process for preparing a benzamide derivative, which comprises reacting a compound represented by the formula (2) and a compound represented by the formula (3) to prepare a compound represented by the formula (1)

[Reaction Scheme 1]

은 본 명세서에서 정의한 바와 같다).
(In the above Reaction Scheme 1, R and

Are as defined herein.

Furthermore, the present invention provides a novel complex labeled with a radioactive isotope of a novel benzamide derivative to which both functional chelating agents represented by the following formula (1) are bonded:

[Chemical Formula 1]

은 본 명세서에서 정의한 바와 같다).
(Wherein R and < RTI ID = 0.0 >

Are as defined herein.

The present invention also provides a pharmaceutical composition for skin cancer diagnosis comprising the above-mentioned complex compound as an active ingredient.

Furthermore, the present invention provides a kit for skin cancer diagnosis comprising the complex compound as an active ingredient.

The novel benzamide derivatives according to the present invention stably form complexes with gallium ( ⁶⁸ Ga), and the complexes produced have excellent selectivity to skin cancer cells and are excreted through the kidneys. Therefore, safe. In addition, since the accumulation rate of skin cancer cells and the accumulation rate of skin cancer cells over time are superior to muscle as a background of an image, and thus nuclear medicine imaging efficiency is excellent, a pharmaceutical composition containing the same as an active ingredient, A basal cell carcinoma, a squamous cell carcinoma, and the like.

FIG. 1 is a graph showing the labeling efficiency in the serum of the complexes prepared in Example 3 according to the present invention over time. FIG.
FIG. 2 is a photograph showing a proton-emission tomography (PET) image of a mouse administered with the complex prepared in Example 3 according to the present invention in a tail vein.

Hereinafter, the present invention will be described in more detail.

The present invention provides a novel benzamide derivative or a pharmaceutically acceptable salt thereof, to which both functional chelating agents represented by the following general formula (1) are bonded:

In Formula 1,

R is - (CH ₂ ) _n -NR ₁ R ₂ , R ₁ and R ₂ are independently of each other hydrogen, C ₁ -C ₄ linear or branched alkyl and n is an integer from 1 to 4; And

는

,

,

또는

이다.

The

,

,

or

to be.

More preferably,

R ₁ and R ₂ are independently of each other methyl or ethyl; n is an integer of 1 to 2; And

는

또는

이다.

The

or

to be.

Most preferably, the amphoteric chelating agent-conjugated benzamide derivative represented by Formula 1 is:

(1) Synthesis of 2,2 ', 2 "- (2- (4- (3- (4- (2- (diethylamino) ethylcarbamoyl) phenyl) thioureido) Triazonan-l, 4,7-triyl) triacetic acid; or

(2) Synthesis of 2,2 ', 2 ", 2''- (2- (4- (3- (4- (2- (diethylamino) ethylcarbamoyl) phenyl) thioureido) benzyl) , 4,7,10-tetraazacyclododecane-1,4,7,10-tetrayl) tetraacetic acid.

The derivative of formula (I) according to the present invention can be used in the form of a pharmaceutically acceptable salt. As the salt, an acid addition salt formed by a pharmaceutically acceptable free acid is useful. Acid addition salts include those derived from inorganic acids such as hydrochloric acid, nitric acid, phosphoric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, nitrous acid or phosphorous acid, and aliphatic mono- and dicarboxylates, phenyl-substituted alkanoates, hydroxyalkanoates, Derived from organic acids such as acetic acid, benzoic acid, citric acid, lactic acid, maleic acid, gluconic acid, methanesulfonic acid, 4-toluenesulfonic acid, tartaric acid, fumaric acid, and the like. Such pharmaceutically innocuous salts include, but are not limited to, sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, nitrate, phosphate, monohydrogenphosphate, dihydrogenphosphate, metaphosphate, pyrophosphate chloride, bromide, Butyrate, caprate, heptanoate, propiolate, oxalate, malonate, succinate, succinate, maleic anhydride, maleic anhydride, , Sebacate, fumarate, maleate, butyne-1,4-dioate, hexane-1,6-dioate, benzoate, chlorobenzoate, methylbenzoate, dynitrobenzoate, hydroxybenzoate, Methoxybenzoate, phthalate, terephthalate, benzenesulfonate, toluene sulfonate, claw Benzenesulfonate, xylenesulfonate, phenylacetate, phenylpropionate, phenylbutyrate, citrate, lactate,? -Hydroxybutyrate, glycolate, maleate, tartrate, methanesulfonate, propanesulfonate, naphthalene- 1-sulfonate, naphthalene-2-sulfonate, mandelate and the like.

The acid addition salt according to the present invention can be obtained by a conventional method, for example, by dissolving a derivative of Chemical Formula 1 in an organic solvent such as methanol, ethanol, acetone, methylene chloride, acetonitrile, , Or may be prepared by drying, or after the solvent and excess acid are distilled off under reduced pressure, followed by drying or crystallization in an organic solvent.

The benzamide derivatives represented by Formula 1 according to the present invention may include not only pharmaceutically acceptable salts, but also all salts, hydrates and solvates which can be prepared by conventional methods.

Also, as shown in the following Reaction Scheme 1,

There is provided a process for preparing a benzamide derivative, which comprises reacting a compound represented by the formula (2) and a compound represented by the formula (3) to prepare a compound represented by the formula (1)

[Reaction Scheme 1]

은 상기 화학식 1에서 정의한 바와 같다).
(In the above Reaction Scheme 1, R and

Is the same as defined in the above formula (1)).

The benzamide derivative represented by formula (1) according to the present invention can be prepared by reacting a compound of formula (2) containing a functional chelating agent with a compound of formula (3) containing a benzamide group in the presence of a base to produce an isothiocyanate And a thiourea produced by adding an amine group of the compound of Formula 3 as a linker.

As the base, there can be used methylamine, ethylamine, dimethylamine, diethylamine, trimethylamine, triethylamine, cyclohexylamine, diethylisopropylamine or pyridine, preferably triethylamine Can be used.

The reaction solution may be dichloromethane, dichloroethane, chloroform, tetrahydrofuran, diethyl ether, toluene, xylene, xylene, benzene, chlorobenzene or dimethylformamide, preferably chloroform Can be used.

Further, the present invention provides a novel complex labeled with a radioisotope to a novel benzamide derivative to which both functional chelating agents represented by the following formula (1) are bonded:

[Chemical Formula 1]

은 상기 화학식 1에서 정의한 바와 같다).
(Wherein R and < RTI ID = 0.0 >

Is the same as defined in the above formula (1)).

In a complex in which a radioactive isotope is labeled with a benzamide derivative having a functional chelating agent bonded to the functional chelate represented by formula (1) according to the present invention, the radioactive isotope may be detected by MRI, CT, A metal containing a transition metal having properties and a radioactive isotope emitting radiation. For example, scandium ⁽⁴⁷ Sc), copper ⁽⁶⁴ Cu), gallium ⁽⁶⁸ Ga), gallium ⁽⁷¹ Ga), yttrium ⁽⁸⁶ Y), yttrium ⁽⁹⁰ Y), technetium ^(99m Tc), indium ⁽¹¹¹ In ), promethium ⁽¹⁴⁹ Pm), samarium ⁽¹⁵³ Sm), dysprosium ⁽¹⁶⁵ Dy), holmium ⁽¹⁶⁶ Ho), erbium ⁽¹⁶⁹ Er), lutetium ⁽¹⁷⁷ Lu), rhenium ⁽¹⁸⁶ Re), rhenium ⁽¹⁸⁸ Re) And bismuth ( ²¹² Bi). And may preferably include gallium ( ⁶⁸ Ga).

In addition, complexes in which a radioactive isotope is labeled with a benzamide derivative having a functional chelating agent represented by the above formula (1) may be prepared from a radioisotope source such as, for example, gallium chloride ( ⁶⁸ GaCl ₃ ) And a benzamide derivative of the formula (1), and reacting the mixture at a temperature of 90 ° C to 110 ° C to produce a radioisotope-labeled complex.

In addition, the present invention provides a pharmaceutical composition for diagnosing skin cancer, which comprises a benzamide derivative to which both functional chelating agents represented by the general formula (1) are conjugated and a radioisotope-labeled complex as an active ingredient.

At this time, the radioisotope includes a metal containing a transition metal having a property of being detectable by MRI, CT, gamma camera or the like and a radioactive isotope releasing radiation when present in a living body. For example, scandium ⁽⁴⁷ Sc), copper ⁽⁶⁴ Cu), gallium ⁽⁶⁸ Ga), gallium ⁽⁷¹ Ga), yttrium ⁽⁸⁶ Y), yttrium ⁽⁹⁰ Y), technetium ^(99m Tc), indium ⁽¹¹¹ In ), promethium ⁽¹⁴⁹ Pm), samarium ⁽¹⁵³ Sm), dysprosium ⁽¹⁶⁵ Dy), holmium ⁽¹⁶⁶ Ho), erbium ⁽¹⁶⁹ Er), lutetium ⁽¹⁷⁷ Lu), rhenium ⁽¹⁸⁶ Re), rhenium ⁽¹⁸⁸ Re) And bismuth ( ²¹² Bi). And may preferably include gallium ( ⁶⁸ Ga).

In addition, the pharmaceutical composition for diagnosing skin cancer according to the present invention can be used for diagnosis of skin cancer such as malignant melanoma, basal cell carcinoma, squamous cell carcinoma and the like, and is preferably used for diagnosis of malignant melanoma. In addition, the pharmaceutical composition for diagnosing skin cancer may be administered by intravenous injection, and the skin cancer may be diagnosed by positron emission tomography (PET).

Since the benzamide derivative represented by the formula (1) according to the present invention contains a bi-functional chelating agent, it is easy to label the radioisotope (see Example 3 and Example 4), and includes a benzamide group having high affinity with melanin Therefore, it is possible to selectively detect melanin-producing skin cancer cells (see Experimental Example 2 and Experimental Example 3).

In addition, the novel benzamide derivatives according to the present invention stably form a complex with gallium ( ⁶⁸ Ga), and after the complex compound labeled with gallium ( ⁶⁸ Ga) becomes a radioisotope label, a 95% (See Experimental Example 1). In addition, since the radiochemical purity is high,

Furthermore, since the complex of the present invention having gallium ( ⁶⁸ Ga) has high solubility in water and is discharged through the kidney after nuclear medicine imaging for skin cancer, it is safe for the human body and is highly selective for skin cancer cells, The accumulation rate over time is excellent and the accumulation rate of skin cancer cells in the background of the image obtained by the diagnostic test is remarkably excellent and the nuclear medicine imaging efficiency of skin cancer is remarkably excellent (see Experimental Example 2 and Experimental Example 3) ).

Therefore, the pharmaceutical composition containing the complex compound in which gallium ( ⁶⁸ Ga) is labeled as an active ingredient in the benzamide derivative having the functional chelating agent bonded to the functional chelate represented by the formula (1) according to the present invention is useful as a malignant melanoma, basal cell carcinoma, And can be usefully used as a pharmaceutical composition for diagnosing skin cancer such as cancer.

When the composition of the present invention is used as a medicine, the pharmaceutical composition containing the complex of the present invention in which gallium ( ⁶⁸ Ga) is labeled or a pharmaceutically acceptable salt thereof as an active ingredient can be formulated and administered at the time of clinical administration However, the present invention is not limited thereto. The pharmaceutical composition may be administered parenterally, and parenteral administration may be by subcutaneous injection, intravenous injection, intramuscular injection, or intra-thoracic injection.

In this case, in order to formulate the formulation for parenteral administration, the complex of the compound labeled with gallium ( ⁶⁸ Ga) or a pharmaceutically acceptable salt thereof is mixed with water or a stabilizer or a buffer to prepare a solution or suspension, and the ampule or vial Unit dosage form. The compositions may contain sterilized and / or preservatives, stabilizers, wettable or emulsifying accelerators, adjuvants such as salts and / or buffers for the control of osmotic pressure, and other therapeutically useful substances, Or may be formulated according to the coating method.

The dose of the pharmaceutical composition according to the present invention to the human body may be varied depending on the patient's age, weight, sex, dosage form, health condition, and disease severity, and the general diagnosis based on adult patients having a body weight of 70 kg 1 to 20 mCi / dose, preferably 3 to 7 mCi / dose. In addition, depending on the judgment of a doctor or a pharmacist, it may be administered once or several times a day at intervals of a certain time. In addition, it is 1 to 1000 mCi / dose, preferably 1 to 500 mCi / dose in the case of treatment, and may be administered once or several times a day at a predetermined time interval according to the judgment of a doctor or pharmacist.

Further, the present invention provides a skin cancer diagnosis kit for detecting skin cancer cells using a complex compound having a radioisotope labeled with a novel benzamide derivative to which both functional chelating agents represented by formula (I) are labeled.

At this time, the radioactive isotope includes a metal including a transition metal having a property of being detectable by MRI, CT, a gamma camera or the like, and a radioactive isotope emitting radiation when the radioactive isotope exists in a living body. For example, scandium ⁽⁴⁷ Sc), copper ⁽⁶⁴ Cu), gallium ⁽⁶⁸ Ga), gallium ⁽⁷¹ Ga), yttrium ⁽⁸⁶ Y), yttrium ⁽⁹⁰ Y), technetium ^(99m Tc), indium ⁽¹¹¹ In ), promethium ⁽¹⁴⁹ Pm), samarium ⁽¹⁵³ Sm), dysprosium ⁽¹⁶⁵ Dy), holmium ⁽¹⁶⁶ Ho), erbium ⁽¹⁶⁹ Er), lutetium ⁽¹⁷⁷ Lu), rhenium ⁽¹⁸⁶ Re), rhenium ⁽¹⁸⁸ Re) And bismuth ( ²¹² Bi). And may preferably include gallium ( ⁶⁸ Ga).

Examples of the skin cancer to which the diagnostic kit according to the present invention can be applied include skin cancer such as malignant melanoma, basal cell skin cancer, and squamous cell skin cancer. Preferably for the diagnosis of malignant melanoma.

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

However, the following Examples and Experimental Examples are merely illustrative of the present invention, and the present invention is not limited to the following Examples and Experimental Examples.

Example 1 Synthesis of 2,2 ', 2 "- (2- (4- (3- (4- (2- (diethylamino) ethylcarbamoyl) phenyl) thioureido) Preparation of 7-triazonan-l, 4,7-triyl) triacetic acid

A solution of 2- (4-isothiocyanatobenzyl) -1,4,7-triazonan-l, 4,7-tricarboxylic acid (0.05 g, 0.09 mmol) Ethyl) benzamide (0.030 g, 0.11 mmol) was dissolved in chloroform (1 ml), followed by the addition of triethylamine (45 μl, 0.27 mmol) and stirring at room temperature for 24 hours. Thereafter, the solvent was distilled off under reduced pressure, and the residue was purified by high performance liquid chromatography (HPLC) using a Phenomenex Luna column to obtain the desired compound (50 mg, 85%).

mp: 180-184 DEG C;

MS (FAB) m / z 686, (M + H) < ⁺ >;

_{HRMS [C 33 H 48 N 7} O 7 S]: 686.3336 ( calculated value) and 686.3340 (measured value);

^{1 H NMR (500 MHz, D} 2 O): δ 7.68 (d, 1H), 7.34 (d, 2H), 7.20 (m, 4H), 3.67-3.65 (br, 2H), 3.54-3.50 (br, 2H ), 3.39-3.29 (br, 4H), 3.20-3.12 (br, 6H), 3.09-2.99 (br, 8H), 2.92-2.52 (br, 3H), 1.20-1.

¹³ C NMR (125 MHz, D ₂ O): δ 179.3, 178.5, 176.3, 174.4, 169.8, 142.0, 136.8, 136.1, 130.3, 130.0, 129.8, 128.8, 128.2, 126.0, 125.6, 124.5. 60.6, 57.8, 57.6, 57.2, 57.1, 52.4, 52.1, 51.6, 50.6, 47.6, 46.6, 46.0, 43.5, 34.9, 33.7, 33.6, 8.2, 8.1.

Example 2 Synthesis of 2,2 ', 2 ", 2' '- (2- (4- (3- (4- (2- (diethylamino) ethylcarbamoyl) phenyl) thioureido) -1,4,7,10-tetraazacyclododecane-1,4,7,10-tetrayl) tetraacetic acid

2,2 ', 2 ", 2' '- (2- (4-isothiocyanatobenzyl) -1,4,7-10-tetraazacyclododecane-1,4,7,10-tetrayl) (0.030 g, 0.21 mmol) was dissolved in chloroform (1 ml), and then triethylamine (30 μl, 0.25 mmol) was added to the solution, , 0.21 mmol), and the mixture was stirred at room temperature for 24 hours. Thereafter, the solvent was removed by distillation under reduced pressure, and the residue was purified by high performance liquid chromatography (HPLC) using a Phenomenex Luna column to obtain the desired compound (42 mg, 75%).

mp: 185-187 [deg.] C;

MS (FAB) m / z 787, (M + H) < ⁺ >;

HRMS [C ₃₇ H ₅₅ N ₈ O ₉ S]: 787.3813 (calculated) and 787.3828 (measured);

^{1 H NMR (500 MHz, D} 2 O): δ 7.60 (d, 2H), 7.33 (d, 2H), 7.14 (br, 4H), 3.58 (br, 2H), 3.52-3.27 (br, 6H), 3.26-3.21 (br, 4H), 3.13 (br, 6H), 3.06-3.02 (br, 6H), 3.0-2.9 (br, 6H), 1.14-1.11 (m, 9H);

^{13 C NMR (125 MHz, D} 2 O): δ 179.4, 179.3, 170.0, 141.9, 136.5, 130.4, 130.1, 128.3, 126.2, 126.0, 124.6, 58.8, 58.6, 57.1, 56.3, 55.6, 55.4, 54.8, 50.7 , 49.8, 47.6, 46.6, 34.9, 32.4, 8.2, 8.1.

≪ Example 3 > ⁶⁸ Ga) 2, 2 "- (2- (4- (3- (4- (2- (diethylamino) ethylcarbamoyl) phenyl) thioureido) benzyl-1,4,7- -1,4,7-triyl) triacetic acid

First, 0.1 M hydrochloric acid was poured into a germanium / gallium ( ⁶⁸ Ge / ⁶⁸ Ga) generator to separate eluted gallium chloride ( ⁶⁸ GaCl ₃ ), and the obtained gallium chloride ( ⁶⁸ GaCl ₃ ) was dissolved in 1 M sodium acetate buffer (pH 5). Then, the compound (30 μg) prepared in Example 1 was mixed with the pH-adjusted gallium chloride ( ⁶⁸ GaCl ₃ ) and reacted at 100 ° C. for 10 minutes. When the reaction was completed, the reaction mixture was cooled to room temperature and purified by high performance liquid chromatography (HPLC) using a Phenomenex Luna column to obtain the desired compound (1.8 mg, 60%).

mp 210-215 [deg.] C;

MS (FAB) m / z 753, (M + H) < ⁺ >;

_{HRMS [C 33 H 45 GaN 7} O 7 S]: 752.2357 ( calculated value) and 752.2350 (measured value);

^{1 H NMR (500 MHz, D} 2 O): δ 7.62 (d, J = 8.9 Hz, 2H), 7.32 (d, J = 8.9 Hz, 2H), 7.18 (m, 4H), 3.70-3.55 (br, 6H), 3.43 (br, 1H), 3.34-3.13 (br, 8H), 3.12-2.99 (br, 6H), 2.96-2.71 (br, 4H), 1.12-1.08 (m, 8H);

^{13 C NMR (125 MHz, D} 2 O): δ 181.1, 147.8, 174.5, 174.2, 169.7, 130.0, 129.9, 129.8, 129.2, 128.4, 128.3, 124.6, 115.0, 65.3, 62.3, 61.9, 52.6, 51.1, 50.7 , 47.9, 47.8, 47.7, 46.7, 34.9, 33.0, 23.1, 8.3, 8.2.

≪ Example 4 > ⁶⁸ Phenyl) thioureido) benzyl) -1, 2, 2 '' - (2- (4- (3- Preparation of 4,7,10-tetraazacyclododecane-1,4,7,10-tetrayl) tetraacetic acid

(1.5 mg, 45%) was obtained in the same manner as in Example 3, except that the compound prepared in Example 2 was used instead of the compound prepared in Example 1 in Example 3, ≪ / RTI >

mp 200-203 [deg.] C;

MS (FAB) m / z 853, (M + H) < ⁺ >;

_{HRMS [C 37 H 52 GaN 8} O 9 S]: 853.2834 ( calculated value) and 853.2842 (measured value);

^{1 H NMR (500 MHz, D} 2 O): δ 7.61 (d, J = 8.6 Hz, 2H), 7.26 (d, J = 8.3 Hz, 2H), 7.18 (d, J = 8.3 Hz, 2H), 7.14 (d, J = 8.6 Hz, 2H), 3.66-3.51 (br, 6H), 3.48-3.37 (br, 4H), 3.34-3.05 (br, 16H), 2.94-2.60 (br, 4H), 1.17-1.12 (m, 9H);

¹³ C NMR (125 MHz, D ₂ O): δ 181.1, 181.0, 179.4, 174.4, 174.2, 174.0, 173.2, 169.8, 141.9, 141.8, 130.4, 130.2, 130.1, 130.0, 129.2, 128.3, 126.3, 126.2, , 124.3, 115.0, 57.7, 56.0, 55.0, 53.5, 51.1, 50.7, 47.9, 47.7, 47.4, 46.7, 34.9, 23.1, 8.3, 8.2, 8.1, 8.0.

<Experimental Example 1> ⁶⁸ Ga) - labeled complexes

The following experiment was conducted to evaluate the stability of gallium ( ⁶⁸ Ga) -labeled complexes according to the present invention.

The stability of the complex prepared in Example 3 was measured by radiochemical purity when mixed with phosphorus serum and cultured at 37 ° C. The radiochemical purity was measured by ITLC-SG (Merck, Germany) for 2 hours at intervals of 30 minutes from the start of the incubation of the serum, considering that the complex compounds used for obtaining the nuclear medicine image were actually labeled and used within 1 hour. Respectively. The radioactivity distribution on the ITLC plate was measured using a TLC-scanner (Bioscan Co. USA), and the results are shown in FIG.

As shown in Fig. 1, it can be seen that the complex compound labeled with gallium ( ⁶⁸ Ga) according to the present invention maintains a stable form. More specifically, the complex of the present invention exhibited a high purity of about 95% or more after mixing with phosphorus serum and culturing at 37 ° C for 2 hours. From this, it can be seen that the complex of the present invention stably retains its label purity for longer than the time required to obtain actual nuclear medicine images after gallium ( ⁶⁸ Ga) is labeled.

Therefore, the novel benzamide derivative according to the present invention stably forms a complex with gallium ( ⁶⁸ Ga), and stably maintains its purity in the human body for the time required for diagnosis, thereby facilitating diagnosis of skin cancer. Therefore, May be useful as a pharmaceutical composition for the diagnosis of skin cancer such as malignant melanoma, basal cell carcinoma, squamous cell carcinoma and the like.

<Experimental Example 2> Evaluation of melanoma accumulation rate by in vivo distribution test

The following experiment was conducted to evaluate the melanoma accumulation rate of gallium ( ⁶⁸ Ga) -labeled complex according to the present invention.

Administration of C57BL / 6 mouse melanoma cell line B16F10 cell line (1 × 10 ⁶ cells) to the right thigh and the mixture was experiment with the tumor transplantation C57BL / 6 mice grew in size of tumors after about 14 days, about 1 cm. The complex (100 mCi) obtained in Example 3 was intravenously administered to the tail vein of tumor-transplanted C57BL / 6 mice. After 60 minutes and 120 minutes, the blood, tumor and organs of the mice were harvested and their weights and radioactivity Respectively. At this time, the measurement results were expressed as a percentage of the injected dose per gram (% ID / g), and the results are shown in Table 1 below.

After 60 minutes
Accumulation rate (% ID / g) After 120 minutes
Accumulation rate (% ID / g) blood 0.46 ± 0.16 0.21 + 0.07 Heart 0.22 + 0.03 0.09 ± 0.01 lungs 0.41 + 0.08 0.21 + 0.02 liver 0.62 + 0.02 0.50 0.07 spleen 0.18 + 0.14 0.19 + 0.03 top 0.20 ± 0.16 0.40 ± 0.05 Leader 0.93 + 0.11 0.62 + - 0.23 kidney 5.09 ± 2.16 3.21 + - 0.32 interest 0.24 0.12 0.07 ± 0.00 muscle 0.20 + 0.13 0.15 + 0.03 bone 0.27 ± 0.05 0.24 + 0.13 brain 0.03 ± 0.00 0.02 ± 0.00 Eyeball 0.20 ± 0.03 0.06 0.02 tumor 1.57 + - 0.66 0.69 + 0.14 skin 0.60 + 0.08 0.25 0.01 Tumor / blood ratio 3.37 ± 0.30 3.49 ± 1.07 Tumor / Muscle Ratio 8.28 ± 1.41 4.73 ± 2.00

As shown in Table 1, it can be seen that the complex compound having gallium ( ⁶⁸ Ga) according to the present invention has a high affinity for tumors and accumulates at a high concentration in melanoma. More specifically, the complex of the present invention showed a low accumulation rate of 1% ID / g or less for other organs and blood after 60 minutes of administration to the mouse, while 1.57 + 0.66% ID / g and 5.09 ± 2.16% ID / g, respectively. After 120 minutes, it was found that the accumulation rate was 3.21 ± 0.32% with respect to the elongation, and the accumulation rate was about 0.7% ID / g in the remaining organ. Furthermore, the tumor accumulation rate to the background muscle of the nuclear medicine image was 8.28 ± 1.41% ID / g. From this, it can be seen that the complex compound having gallium ( ⁶⁸ Ga) according to the present invention has a high affinity for the tumor and accumulates in the tumor at a high concentration after being administered to the body. Also, since the solubility in water is excellent, The complexes accumulated in the tumor and the small amount of the complex remaining in the body are discharged through the kidney.

Therefore, the novel benzamide derivative according to the present invention stably forms a complex with gallium ( ⁶⁸ Ga), and the complex is excellent in the selective accumulation rate to skin cancer cells and is excellent in water solubility, All are excreted through the kidneys and are therefore safe for the human body. In addition, since the accumulation rate of skin cancer cells to muscle is excellent, the nuclear medicine diagnostic imaging efficiency is excellent. Therefore, the pharmaceutical composition containing it as an active ingredient is a pharmaceutical composition for diagnosing skin cancer such as malignant melanoma, basal cell cancer, . &Lt; / RTI &gt;

<Experimental Example 3> Positron emission tomography (PET) evaluation

The following experiments were conducted to evaluate the nuclear imaging diagnostic efficiency and safety of positron emission tomography (PET) of the complexes of gallium ( ⁶⁸ Ga) labeled according to the present invention.

(100 mCi) in the tail vein of the tumor transplantation C57BL / 6 mouse model transplanted with the B16F10 cell line at the right femur at the time of 30 minutes, 60 minutes, and 120 minutes after positron emission tomography PET images were obtained. The results are shown in Fig.

As shown in FIG. 2, the complex compound labeled with gallium ( ⁶⁸ Ga) according to the present invention can be usefully used for positron emission tomography (PET) for diagnosis of skin cancer. More specifically, the complexes of the present invention were accumulated in skin tumor cells over time after 30 minutes of tail vein administration, and the position and size of skin cancer were more accurately photographed. From this, it can be seen that the complex compound according to the present invention can obtain accurate nuclear medicine diagnostic images when diagnosing skin cancer.

Therefore, the novel benzamide derivative according to the present invention stably forms a complex with gallium ( ⁶⁸ Ga), and the prepared complex has excellent selective accumulation rate on skin cancer cells, It is safe for human body since it is discharged. In addition, since the accumulation rate of skin cancer cells to muscles and the accumulation rate according to the elapsed time of administration are excellent, the position and size of skin cancer cells can be more accurately diagnosed. Therefore, a pharmaceutical composition containing the same as an active ingredient can be used as a malignant melanoma, Cell cancer, squamous cell cancer, and the like.

Claims (13)

A novel benzamide derivative or a pharmaceutically acceptable salt thereof, to which both functional chelating agents represented by the following formula (1) are bonded:
[Chemical Formula 1]

(In the formula 1,
R is - (CH ₂ ) _n -NR ₁ R ₂ , R ₁ and R ₂ are independently of each other hydrogen, C ₁ -C ₄ linear or branched alkyl and n is an integer from 1 to 4; And

The

,

,

or

to be).
The method according to claim 1,
R ₁ and R ₂ are independently of each other methyl or ethyl; n is an integer of 1 to 2; And

The

or

&Lt; / RTI &gt;
The method according to claim 1,
The amphoteric chelating agent-conjugated benzamide derivatives represented by the above formula (1) are:
(1) Synthesis of 2,2 ', 2 "- (2- (4- (3- (4- (2- (diethylamino) ethylcarbamoyl) phenyl) thioureido) Triazonan-l, 4,7-triyl) triacetic acid; or
(2) Synthesis of 2,2 ', 2 ", 2''- (2- (4- (3- (4- (2- (diethylamino) ethylcarbamoyl) phenyl) thioureido) benzyl) , 4,7,10-tetraazacyclododecane-1,4,7,10-tetrayl) tetraacetic acid.
As shown in Scheme 1 below,
A process for preparing a benzamide derivative according to claim 1, which comprises reacting a compound represented by the formula (2) and a compound represented by the formula (3) to prepare a compound represented by the formula (1)
[Reaction Scheme 1]

(In the above Reaction Scheme 1, R and

Lt; / RTI &gt; are as defined in claim 1).
12. A complex of a novel benzamide derivative or a pharmaceutically acceptable salt thereof conjugated with a functional chelator of claim 1 labeled with a radioisotope.
6. The method of claim 5,
Wherein the radioactive isotope is gallium ( ⁶⁸ Ga).
A pharmaceutical composition for diagnosing skin cancer, which contains, as an active ingredient, a novel benzamide derivative having a functional chelating agent represented by the following formula (1) or a pharmaceutically acceptable salt thereof and a complex having a radioisotope labeled:
[Chemical Formula 1]

(Wherein R and &lt; RTI ID = 0.0 &gt;

Lt; / RTI &gt; are as defined in claim 1).
8. The method of claim 7,
Wherein the radiation isotope is gallium ( ⁶⁸ Ga).
8. The method of claim 7,
Wherein the skin cancer is one kind of skin cancer selected from the group consisting of malignant melanoma, basal cell skin cancer and squamous cell skin cancer.
8. The method of claim 7,
Wherein said pharmaceutical composition is administered by intravenous injection, and then diagnosed with skin cancer by positron emission tomography (PET).
A skin cancer diagnosis kit for detecting skin cancer cells using a novel benzamide derivative having a functional chelating agent represented by the following formula (1) or a pharmaceutically acceptable salt thereof and a complex labeled with a radioisotope:
[Chemical Formula 1]

(Wherein R and &lt; RTI ID = 0.0 &gt;

Lt; / RTI &gt; are as defined in claim 1).
12. The method of claim 11,
Wherein the radiation isotope is gallium ( ⁶⁸ Ga).
12. The method of claim 11,
Wherein the skin cancer is one kind of skin cancer selected from the group consisting of malignant melanoma, basal cell cancer and squamous cell cancer.

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