Radioactive compounds for the treatment of melanoma and use thereof

Abstract

The present invention relates to a radioactive compound having improved targeting ability to melanoma and a pharmaceutical composition for treating melanoma comprising the same, which has improved targeting ability to melanoma compared to a conventional melanoma contrast agent. The radioactive compound according to the present invention embodiment shows a very excellent therapeutic effect to melanoma.

Description

[0001] This application claims priority to Korean Patent Application No. 2018-0074766, filed on June 28, 2018, the entire contents of which are incorporated herein by reference. Technical Field

[0002] This invention relates to a novel compound and its use, and more specifically, to a novel radioactive compound for the treatment of melanoma and its use. Background Technology

[0003] Malignant melanoma is considered one of the deadliest cancers due to its high rate of metastasis throughout the body. While it accounts for 5% of all skin cancers, it is responsible for over 50% of skin cancer-related deaths. Furthermore, the incidence of this disease has doubled in the past two decades and continues to rise steadily. To date, no effective treatment has been developed for melanoma. However, early diagnosis and accurate determination of the disease stage are considered crucial for improving survival rates for patients with malignant melanoma.

[0004] recent, 18 FN-[2-(diethylamino)ethyl]-4-fluoro-benzamide ( 18 F-FBZA was developed and its application as a PET contrast agent targeting melanin in the detection of metastatic melanoma has been reported (Ren et al., J. Nucl. Med. 50(10): 1692-1699, 2009). However, when used as a PET contrast agent for melanoma detection, the principle is to induce selective uptake by melanoma through the chemical transformation of the benzamide structure. However, the uptake rate of melanoma is low and the image quality is poor, thus requiring improvement. Furthermore, until now, the only treatment option for melanoma is surgical resection of the lesion and the use of anticancer drugs. However, surgery has many location limitations, the drugs used have the disadvantage of risk of recurrence, and the treatment response is very low. Therefore, radiotherapy by selectively uptake of radioactive compounds by malignant melanoma is considered the most effective method, but an effective radiopharmaceutical targeting melanoma has not yet been developed. Summary of the Invention

[0005] Technical issues

[0006] The present invention aims to solve various problems, including those mentioned above, and the object of the present invention is to provide novel radiotherapy compounds with improved melanoma targeting capabilities.

[0007] Another object of the present invention is to provide a pharmaceutical composition for treating melanoma, comprising the said compound.

[0008] However, these problems are exemplary, and the scope of the invention is not limited thereto.

[0009] Technical solution

[0010] In one aspect of the invention, a novel radioactive compound or an acceptable salt thereof is provided, having a structure as shown in Formula 1 or Formula 2:

[0011]

[0012] (In the above formula, X1 and X2 are each independently carbon or nitrogen, at least one of which is nitrogen; L1 is absent or chemically bonded, or is a substituted or unsubstituted alkylene group having 1 to 20 carbon atoms, a substituted or unsubstituted allyl group having 6 to 14 carbon atoms, a substituted or unsubstituted heteroalkylene group having 1 to 20 carbon atoms, a (poly)alkylene glycol having 2 to 60 carbon atoms, or one or more connectors selected from the following) L2 is a chemically bonded, substituted, or unsubstituted alkylene group having 1 to 5 carbon atoms; R1 is selected from... 80m Br、 123 I, 124 I, 125 I, 131 I, and 32 The radioactive isotope of P or a functional group containing a radioactive isotope, and L is DOTA (1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid), DOTA-NCS, DOTA-NHS ester, DOTA-Bz-NCS, tris(tert-butyl)DOTA, HBED-CC-TFP ester, DTPA (diethylenetriaminepentaacetic acid), DO3A (1,4,7,10-tetraazacyclododecane-1,4,7-triacetic acid), NOTA (1,4,7-triazacyclononane-1,4,7-triacetic acid), NODAGA (1,4,7... - Triazacyclononane, 1-glutaric acid-4,7-acetic acid), TETA (1,4,8,11-tetraazatetradecane-N,N',N″,N″′-tetraacetic acid), TE3A (1,4,8,11-tetraazatetradecane-1,4,8-triacetic acid), TE2A (1,4,8,11-tetraazabicyclohexadecane-4,11-diacetic acid), PCTA (3,6,9,15-tetraazabicyclo[9.3.1]pentadecane-1,11,13-triene-3,6,9,-triacetic acid), macrocyclic polyamines, cyclop-amines, or DFO (deferroamine); M is a radioactive metal selected from... 64 Cu、 67 Cu、 90 Cu、 68 Ga、99 mTc, 85 Sr、 89 Sr、 86 Y、 90 Y、 99 mTc, 111 In、 114m In、 149 Tb, 152 Tb, 153 Sm、 165 Dy、 166 Ho、 169 Er、 177 Lu、 186 Re、 188 Re、 198 Au、 211 At、 212 Pb, 223 Ra、 224 Ra、 225 Ac and 255 Fm is a radioactive metal; R2 and R3 are each independently hydrogen, hydroxyl, alkyl with 1-3 carbon atoms, acetamido, or alkoxy with 1-3 carbon atoms.

[0013] Another aspect of the present invention provides a pharmaceutical composition for treating malignant melanoma, comprising a compound of formula 1 or 2 or a pharmaceutically acceptable salt thereof as an active ingredient.

[0014] Beneficial effects

[0015] A novel radioactive compound according to one embodiment of the present invention, and its pharmaceutically acceptable salt, can be used as a therapeutic agent for melanoma. The radioactive compound according to an embodiment of the present invention not only has improved targeting ability against melanoma, but also has the advantage of significantly inhibiting melanoma growth. However, the scope of the present invention is not limited to the above-described effects. Attached Figure Description

[0016] Figure 1 A chromatogram showing the results of the analysis of the labeled yield and radiochemical purity was provided, obtained by separating the radioactive compound according to an embodiment of the invention using radioactive thin-layer chromatography.

[0017] Figure 2 To illustrate, when an embodiment of the present invention is presented... 177 A graph showing the change in tumor growth over time when Lu-DOTA-NCS-DMPY2 is applied to a small animal model of melanoma.

[0018] Figure 3 To illustrate when one embodiment of the present invention is 177A graph showing the change in body weight over time when Lu-DOTA-NCS-DMPY2 is applied to a small animal model of melanoma. Detailed Implementation

[0019] One aspect of the present invention provides novel radioactive compounds having a structure of formula 1 or 2, or pharmaceutically acceptable salts thereof.

[0020]

[0021] (In the above formula, X1 and X2 are each independently carbon or nitrogen, at least one of which is nitrogen; L1 is absent or chemically bonded, or is a substituted or unsubstituted alkylene group having 1 to 20 carbon atoms, a substituted or unsubstituted allyl group having 6 to 14 carbon atoms, a substituted or unsubstituted heteroalkylene group having 1 to 20 carbon atoms, a (poly)alkylene glycol having 2 to 60 carbon atoms, or one or more connectors selected from the following) L2 is a chemically bonded, substituted, or unsubstituted alkylene group having 1 to 5 carbon atoms; and R1 is selected from... 80m Br、 123 I, 124 I, 125 I, 131 I, and 32 The radioactive isotope of P or a functional group containing a radioactive isotope, and L is DOTA (1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid), DOTA-NCS, DOTA-NHS ester, DOTA-Bz-NCS, tris(tert-butyl)DOTA, HBED-CC-TFP ester, DTPA (diethylenetriaminepentaacetic acid), DO3A (1,4,7,10-tetraazacyclododecane-1,4,7-triacetic acid), NOTA (1,4,7-triazacyclononane-1,4,7-triacetic acid), NODAGA (1,4,7... - Triazacyclononane, 1-glutaric acid-4,7-acetic acid), TETA (1,4,8,11-tetraazatetradecane-N,N',N″,N″′-tetraacetic acid), TE3A (1,4,8,11-tetraazatetradecane-1,4,8-triacetic acid), TE2A (1,4,8,11-tetraazabicyclohexadecane-4,11-diacetic acid), PCTA (3,6,9,15-tetraazabicyclo[9.3.1]pentadecane-1,11,13-triene-3,6,9,-triacetic acid), macrocyclic polyamines, cyclop-amines, or DFO (deferroamine); M is a radioactive metal selected from... 64 Cu、 67 Cu、 90 Cu、 68 Ga、 99 mTc, 85Sr、 89 Sr、 86 Y、 90 Y、 99 mTc, 111 In、 114m In、 149 Tb, 152 Tb, 153 Sm、 165 Dy、 166 Ho、 169 Er、 177 Lu、 186 Re、 188 Re、 198 Au、 211 At、 212 Pb, 223 Ra、 224 Ra、 225 Ac and 255 Fm is a radioactive metal; R2 and R3 are each independently hydrogen, hydroxyl, alkyl with 1-3 carbon atoms, acetamido, or alkoxy with 1-3 carbon atoms.

[0022] In the novel radioactive compound or an acceptable salt thereof, in the case of Formula 1 and R1, and in the case of Formula 2, the [M←L] portion is a radioactive isotope portion, and L1 is a connector portion connecting the radioactive isotope portion and the melanoma target portion, and the R1 or right structure excluding the [M←L] portion and L1 corresponds to the melanoma target portion.

[0023] In the novel radioactive compound or its pharmaceutically acceptable salt, the linker L1 can be a modular linker that links the chelating agent to the melanoma-targeting moiety via click chemistry, such as azide-alkyne cycloaddition reactions, in addition to general organic synthesis, click chemistry reactions of alkyne-nitroso, (alkyne-nitro cycloaddition) reactions, olefin and azide [3+2] cycloaddition reactions, olefin and tetrazine reverse demand Diels-Alder reactions, and in addition to general organic synthesis, photoclick reactions of alkyne and tetrazine to form triazoles or dibenzotriazoloazocine. The modular linker may include one or more functional linkers, including hydrocarbon chain connectors (-[CH2]). n -) or polyethylene glycol (-[C2H4O]n-), which is used as a spacer group to separate the radioactive isotope portion from the melanoma targeting portion at a certain distance, and a hydrophobic portion (e.g., ) used to reduce the hydrophilicity of radioactive compounds or their pharmaceutically acceptable salt or albumin-binding moieties (e.g., It is used to enhance the in vitro stability of radioactive compounds or pharmaceutically acceptable salts, and is added in a modular manner.

[0024] In the novel radioactive compound or its pharmaceutically acceptable salt, the chemical bond may be an ester bond, amide bond, ether bond, thioether bond, thioester bond, or disulfide bond, and L2 may more preferably be an alkylene group (-(CH2)) having 1 to 5 carbon atoms. n - (N is an integer from 1 to 5), most preferably ethylidene (-CH2CH2-) or propylidene (-CH2CH2CH2-).

[0025] Another aspect of the present invention provides a contrast agent for treating melanoma, comprising a compound of formula 1 or 2 or a pharmaceutically acceptable salt thereof as an active ingredient.

[0026] The compounds of Formula 1 can be prepared by the following method:

[0027]

[0028] Furthermore, once a radioactive compound containing a common halogen element rather than a radioactive halogen element is synthesized, salts containing radioactive halogen elements can be used, for example... 80m Br and 123 I is prepared by substitution reaction to produce R1. Alternatively, radioisotope halogen or radiophosphorus can be introduced by reaction with a compound having a radioisotope halogen or radiophosphorus as a functional group (e.g., phosphate group).

[0029] The specific implementation scheme of the compound or its pharmaceutically acceptable salt is as follows:

[0030] (5-(((2-(dimethylamino)ethyl)carbamoyl)pyridin-2-yl)carbamoic acid[ 131 I] Iodide

[0031]

[0032] N-(2-(dimethylamino)ethyl)-6-([ 125 I] Iodomethyl) Nicotinamide

[0033]

[0034] N-(2-(dimethylamino)ethyl)-5-([ 125 I]Iodomethyl)pyridineamide

[0035]

[0036] N-(3-(dimethylamino)propyl)-5-[131 I] Iodopyridine-2-carboxamide

[0037]

[0038] (S)-5-(2-amino-3-(4-hydroxy-3-[ 131 I]Iodophenyl)propamido)-N-(2-(dimethylamino)ethyl)pyridinecarboxamide

[0039]

[0040] (S)-5-(4-(4-(2-(2-amino-3-(4-hydroxy-3-[ 125 I]Iodophenyl)propamido)ethyl)-1H-1,2,3-triazol-1-yl)butamido)-N-(2-(dimethylamino)ethyl)pyridinecarboxamide

[0041]

[0042] (S)-6-(6-(3-(3-(2-amino-3-(4-hydroxy-3-[125I]iodophenyl)propamido)propyl)-3H-dibenzo[b,f][1,2,3]triazolo[4,5-d]azo-8(9H)-yl)-6-oxohexamido)-N-(2-(dimethylamino)ethyl)nicotinamide

[0043]

[0044] (6-((2-(dimethylamino)ethyl)carbamoyl)pyridin-3-yl)methyldihydro[ 32 (P] phosphate ester)

[0045]

[0046] 4-Acetamido-N-(2-(dimethylamino)ethyl)-5-[ 131 I] Iodine-2-methoxybenzamide

[0047]

[0048] 177 Lu-DOTA-DMP

[0049]

[0050] 177 Lu-DOTA-DMPY2

[0051]

[0052] 177Lu-DOTA-NCS-DMP

[0053]

[0054] 177 Lu-DOTA-NCS-DMPY2

[0055]

[0056] 177 Lu-DOTA-NCS-triazole-PEG-DMP

[0057]

[0058] 177 Lu-DOTA-NCS-triazole-PEG-DMPY2

[0059]

[0060] 177 Lu-DOTA-NCS-ADIBO-DMP

[0061]

[0062] 177 [[ID=�5]]Lu-DOTA-NCS-ADIBO-DMPY2

[0063]

[0064] 177 Lu-DOTA-triazole-DMP

[0065]

[0066] 177 Lu-DOTA-triazole-DMPY2

[0067]

[0068] 177 Lu-DOTA-triazole-PEG-DMP<००००३२०>

[0069]

[0070] 177 Lu-DOTA-triazole-PEG-DMPY2

[0071]

[0072] 177 Lu-DOTA-ADIBO-DMP It should be noted that in the translation, for the 7-digit tags like ,

[0053] , etc., they are kept exactly as they are according to the requirements. And for the chemical compound names, they are translated as accurately as possible while maintaining the original format. There seems to be an error in the tag <००००३२०> in the original text which should probably be and is translated accordingly.

[0073]

[0074] 177 Lu-DOTA-ADIBO-DMPY2

[0075]

[0076] In formulas 1 to 25, DOTA is an abbreviation for 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid, NCS represents isocyanate, PEG is an abbreviation for polyethylene glycol, DMP is an abbreviation for 4-amino-N-(3-(dimethylamino)propyl)benzylamide, DMPY2 is an abbreviation for 6-amino-N-(3-(dimethylamino)propyl)nicotinamide, and ADIBO is an abbreviation for azadibenzocyclooctyne.

[0077] On the other hand, the compound of formula 2 can be prepared by first preparing a chelating agent-binding compound as shown in scheme 2 below, and then adding a radioactive isotopic metal element thereto:

[0078]

[0079] As shown in Scheme 1 above, the parent compound can be reacted with hexamethylditine to replace the halogen atom with a trimethyltin group, and then reacted with a halogen compound containing a radioactive isotope. In this case, R1 is the radioactive isotope portion, L1 corresponds to the connector linking the radioactive isotope portion and the melanoma-targeting portion, and the right-hand structure other than R1 and L1 corresponds to the melanoma-targeting portion. Similarly, as shown in Scheme 2, the chelating agent portion can be prepared by binding to the melanoma-targeting portion, and the isothiocyanate (NCS) or succinimide group, which is the reactive group of the chelating agent portion, can be linked to the amino group of the melanoma-targeting portion via a thiourea bond or an amide bond, respectively. The reactive group and bonding method are exemplary and not limited to Scheme 2.

[0080] In scheme 1 or 2, the connector L1 can be a modular connector that links the chelating agent to the melanoma targeting site via click chemistry, such as an azide-alkyne cycloaddition reaction, or click chemistry involving alkyne-nitroso reactions (alkyne-nitro cycloaddition), olefin and azide [3+2] cycloaddition reactions, olefin and tetrazine reverse demand Diels-Alder reactions, and, in addition to general organic synthesis, alkyne and tetrazine photoclick reactions to form triazoles or dibenzotriazoloazocine. The modular connector may include one or more functional connectors, including a hydrocarbon linker (-[CH2]). n-) or polyethylene glycol (-[C2H4O]n-), which is used as a spacer group to separate the radioactive isotope portion from the melanoma targeting portion at a certain distance, and a hydrophobic portion (e.g., ), used to reduce the hydrophilicity of radioactive compounds or their pharmaceutically acceptable salt or albumin-binding moieties (e.g., It is used to enhance the in vivo stability of radioactive compounds or pharmaceutically acceptable salts, and is added in a modular manner.

[0081] In the novel radioactive compound or its pharmaceutically acceptable salt, the chemical bond may be an ester bond, amide bond, ether bond, thioether bond, thioester bond or disulfide bond, and L2 may more preferably be an alkylene group (-(CH2)) having 1 to 5 carbon atoms. n - (N is an integer from 1 to 5), most preferably ethylidene (-CH2CH2-) or propylidene (-CH2CH2CH2-).

[0082] The chelating agent shown in Scheme 2 is exemplary, and various chelating agents described above can be used.

[0083] "Pharmaceutically acceptable salts" are preferably salts of inorganic or organic acids, more preferably salts of aliphatic acids such as methoxy, acetoxy, trifluoroacetoxy anions, chlorides, bromides, iodides, or aromatics, or salts such as aryl aliphatic carboxylates, nitrates, sulfates, phosphates, sulfonates, methanesulfonates, benzenesulfonates, and toluenesulfonates, but are not limited thereto. Furthermore, pharmaceutically acceptable salts of the present invention include salts using F-, Cl-, Br-, or I-. However, pharmaceutically acceptable salts of the present invention are not limited thereto.

[0084] According to another aspect of the invention, a pharmaceutical composition for treating melanoma is provided, comprising the radioactive compound or a pharmaceutically acceptable salt thereof as an active ingredient.

[0085] In practical use, according to conventional pharmaceutical preparation techniques, a pharmaceutical composition according to one embodiment of the present invention can be combined with a pharmaceutically acceptable carrier. Depending on the desired formulation, such as oral or parenteral administration (including intravenous administration), the carrier can take various forms.

[0086] Furthermore, the pharmaceutical composition according to one embodiment of the present invention can be administered at a dose of 0.1 mg / kg to 1 g / kg, more preferably 0.1 mg / kg to 500 mg / kg. Alternatively, the dose can be appropriately adjusted according to the patient's age, sex, and condition within the permissible daily or annual radiation exposure range.

[0087] A pharmaceutical composition according to one embodiment of the invention further includes an inert component, said inert component comprising a pharmaceutically acceptable carrier. As used herein, a "pharmaceutically acceptable carrier" refers to a composition other than the active ingredient, particularly a component of a pharmaceutical composition. Examples of such pharmaceutically acceptable carriers include binders, disintegrants, diluents, fillers, lubricants, solubilizers or emulsifiers, and salts.

[0088] The novel pharmaceutical composition can be administered to the subject via parenteral administration, and parenteral administration can be intravenous injection, intraperitoneal injection, intramuscular injection or subcutaneous administration, but intravenous administration is the most preferred.

[0089] The invention will be described in more detail below with reference to embodiments and experimental examples. However, the invention is not limited to the embodiments disclosed below, but can be implemented in various different forms, and the embodiments are provided to enable those skilled in the art to fully understand the scope of the invention.

[0090] Example 1: Preparation of the precursor

[0091] 1-1: Preparation of DOTA-DMP

[0092]

[0093] 5 g of 4-aminobenzoic acid and 0.987 g of N,N,N',N'-tetramethyl-O-(N-succinimide)urea tetrafluoroborate (TSTU) were dissolved in N,N-dicarboxyformamide (DMF), and 1.72 mL of N,N-diisopropylethylamine (DIPEA) was added. The mixture was then stirred under reflux at 60 °C for 3 hours. After stirring for 3 hours, 0.612 mL of N,N-dimethylethylenediamine (DMEDA) was added, and the mixture was stirred at room temperature for 2 hours. The product was extracted with 60 mL of CH2Cl2 and 130 mL of H2O, and the water in the CH2Cl2 layer was removed with MgSO4 and filtered. The filtrate was evaporated to dryness, and the product 4-amino-N-(2-(dimethylamino)ethyl)benzamide was separated and purified by column chromatography.

[0094] Subsequently, 0.08 g of 4-amino-N-(2-(dimethyl-amino)ethyl)benzamide and 0.229 g of DOTA-NHS ester prepared above were dissolved in CHCl3, and the pH was adjusted to 9-10 using triethylamine. The mixture was then stirred at room temperature for 24 hours. After removing the solvent by vacuum distillation, 2,2',2″-(10-(2-(4-(2-(2-(dimethylamino))ethyl-carbamoyl)phenylamino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid with the structure of Formula 26 above was separated using a semi-preparative column, which is designated as "DOTA-DMP". The NMR data are as follows:

[0095] 1 H-NMR(300MHz,D2O):2.92(s,6H),3.13(br,16H),3.33(t,2H),3.74(t,2H),3.87(br,8H),6.69(d,2H),7.66(d,2H).

[0096] 1-2: Preparation of DOTA-DMPY2

[0097]

[0098] 0.6 g of 5-aminopyridine-2-carboxylic acid and 1.308 g of TSTU were dissolved in DMF, and 2.41 mL of DIPEA was added. The mixture was then stirred at 60 °C for 3 hours under reflux. After stirring for 3 hours, 0.808 mL of DMEDA was added, and the mixture was stirred at room temperature for 2 hours. The product was extracted with 80 mL of CH2Cl2 and 130 mL of H2O, and the water in the CH2Cl2 layer was removed with MgSO4 and filtered. The filtrate was evaporated to dryness, and 5-amino-N-(2-(dimethylamino)ethyl)pyridinecarboxamide (NH2-DMPY2) was separated and purified by column chromatography.

[0099] Subsequently, 0.07 g of NH2-DMPY2 and 0.2 g of DOTA-NHS ester prepared above were dissolved in CHCl3, and the pH was adjusted to 9-10 using triethylamine. The mixture was then stirred at room temperature for 24 hours. Subsequently, distillation was performed under reduced pressure to remove the solvent. Then, 2,2',2″-(10-(2-((6-((((2-(dimethylamino)ethyl))carbamoyl)pyridin-3-yl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid with the structure of Formula 27 was separated using a semi-preparative column, yielding "DOTA-DMPY2". The 1H-NMR analysis results of DOTA-DMPY2 are as follows:

[0100] 1H-NMR(300MHz,D2O):2.99(s,6H),3.16(br,16H),3.41(t,2H),3.77(t,2H),3.85(br,8H),7.93(d,1H),8.35(m,1H),8.88(d,1H).

[0101] 1-3: Preparation of DOTA-NSC-DMP

[0102]

[0103] 0.08 g of NH2-DMFB prepared in Example 1-1 and 0.208 g of 2-(4-isothiocyanate benzyl)-1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (p-SCN-Bn-DOTA) were dissolved in CHCl3, and the pH was adjusted to 9-10 using triethylamine. The mixture was then stirred at room temperature for 24 hours. After removing the solvent by vacuum distillation, 2,2',2″,2″′-(2-(4-(3-(4-(((2-(dimethylamino)ethyl)carbamoyl)phenyl)thioureoyl)benzyl)-1,4,7,10-tetraazacyclododecane-1,4,7,10-tetrayl)tetraacetic acid was separated using a semi-preparative column chromatography, yielding "DOTA-NCS-DMP". DOTA-NCS-DMP... 1 The H-NMR data are as follows:

[0104] 1 H-NMR(300MHz,D2O):2.92(s,6H),3.13(br,16H),3.33(t,2H),3.74(t,2H),3.87(br,8H),6.43(d,2H),6.69(d,2H),6.86(d,2H),7.66(d,2H).

[0105] 1-4: Preparation of DOTA-NCS-DMPY2

[0106]

[0107] 0.07 g of NH2-DMPY2 prepared in Examples 1-2 and 0.181 g of p-SCN-Bn-DOTA were dissolved in CHCl3, and the pH was adjusted to 9-10 using triethylamine. The mixture was then stirred at room temperature for 24 hours. After removing the solvent by vacuum distillation, 2,2',2″,2″′-(2-(4-(3-(6-(((2-(di(methylamino)ethyl)carbamoyl))pyridin-3-yl)thiourea)benzyl)-1,4,7,10-tetraaza-cyclododecane-1,4,7,10-tetrayl)tetraacetic acid was separated using a semi-preparative column chromatography, yielding "DOTA-NCS-DMPY2". The DOTA-NCS-DMPY2... 1 The H-NMR data are as follows:

[0108] 1 H-NMR(300MHz,D2O):2.99(s,6H),3.16(br,16H),3.41(t,2H),3.77(t,2H),3 .85(br,8H),6.42(d,2H),6.87(d,2H),7.93(d,1H),8.35(m,1H),8.88(d,1H).

[0109] Example 2: Preparation of Standard Materials

[0110] 2-1: Preparation of Lu-DOTA-DMP

[0111]

[0112] 5 mg of DOTA-DMP prepared in Example 1-1 and 5 mg of LuCl3 were dissolved in 0.2 M sodium acetate buffer and stirred at 95 °C for 1 hour. Lu-DOTA-DMP having the structure of Formula 30 above was separated using a semi-preparative column. The structure of Lu-DOTA-DMP... 1 The H-NMR data are as follows:

[0113] 1 H-NMR(300MHz,D2O):2.94(s,6H),3.12(br,16H),3.34(t,2H),3.72(t,2H),3.85(br,8H),6.68(d,2H),7.65(d,2H).

[0114] 2-2:Preparation of Lu-DOTA-DMPY2

[0115]

[0116] 7 mg of DOTA-DMPY2 prepared in Examples 1-2 and 7 mg of LuCl3 were dissolved in 0.2 M sodium acetate buffer and stirred at 95 °C for 1 hour. Lu-DOTA-DMPY2 having the structure of Formula 31 above was separated using a semi-preparative column. The structure of Lu-DOTA-DMPY2... 1 The H-NMR data are as follows:

[0117] 1H-NMR(300MHz,D2O):2.98(s,6H),3.14(br,16H),3.38(t,2H),3.79(t,2H),3.88(br,8H),7.91(d,1H),8.38(m,1H),8.90(d,1H).

[0118] 2-3: Preparation of Lu-DOTA-NCS-DMP

[0119]

[0120] 5 mg of DOTA-NCS-DMP prepared in Examples 1-3 and 4 mg of LuCl3 were dissolved in 0.2 M sodium acetate buffer and stirred at 95 °C for 1 hour. Lu-DOTA-NCS-DMP having the structure of Formula 32 above was separated using a semi-preparative column. The structure of Lu-DOTA-NCS-DMP... 1 The H-NMR data are as follows:

[0121] 1 H-NMR(300MHz,D2O):2.94(s,6H),3.12(br,16H),3.34(t,2H),3.72(t,2H),3.85(br,8H),6.45(d,2H),6.68(d,2H),6.85(d,2H),7.65(d,2H).

[0122] 2-4: Preparation of Lu-DOTA-NCS-DMPY2

[0123]

[0124] 7 mg of DOTA-NCS-DMPY2 prepared in Examples 1-4 and 6 mg of LuCl3 were dissolved in 0.2 M sodium acetate buffer solution and stirred at 95 °C for 1 hour. Lu-DOTA-NCS-DMPY2 having the structure of Formula 33 above was separated using a semi-preparative column. The Lu-DOTA-NCS-DMPY2... 1 The H-NMR data are as follows:

[0125] 1H-NMR(300MHz,D2O):2.98(s,6H),3.14(br,16H),3.38(t,2H),3.79(t,2H),3 .88(br,8H),6.43(d,2H),6.89(d,2H),7.91(d,1H),8.38(m,1H),8.90(d,1H).

[0126] Example 3: Preparation of radioactive compounds

[0127] 3-1: Preparation 177 Lu-DOTA-DMP

[0128]

[0129] 30 μg of DOTA-DMP prepared in Example 1-1 and 177 LuCl3 (20 mCi) was dissolved in 0.2 M sodium acetate buffer solution, and the reaction mixture was incubated at 90 °C for 1 hour to synthesize a compound having the structure of Formula 12 above. 177 Lu-DOTA-DMP. After cooling the reaction mixture at room temperature, the radioactive compound was separated and purified using a semi-preparative column.

[0130] 3-2: 177 Preparation of Lu-DOTA-DMPY2

[0131]

[0132] 30 μg of DOTA-DMPY2 prepared in Examples 1-2 and 177 LuCl3 (30 mCi) was dissolved in 0.2 M sodium acetate buffer solution, and the reaction mixture was incubated at 90 °C for 1 hour to synthesize a compound having the structure of Formula 13 above. 177 Lu-DOTA-DMPY2. After cooling the reaction mixture at room temperature, the radioactive compound was separated and purified using a semi-preparative column.

[0133] As mentioned above, the radioactive isotope ruthenium will be used. 177 Lu-marked 177 Lu-DOTA-DMPY2 was separated using radiometric thin-layer chromatography with an aqueous citric acid solution as the developing solvent to analyze the labeling yield and radiochemical purity. For example... Figure 1 As shown, peaks appearing between 20-40 mm in the development distance indicate peaks synthesized through embodiments of the present invention. 177 Lu-DOTA-DMPY2, while peaks appearing between 85-95 mm in the development distance indicate unlabeled free peaks. 177 Lu. The results of peak area calculations confirmed the above synthesis. 177 The radiochemical purity of Lu-DOTA-DMPY2 is >98% or higher.

[0134] 3-3: Preparation of 177Lu-DOTA-NCS-DMP

[0135]

[0136] 30 μg of DOTA-NCS-DMP prepared in Examples 1-3 and 177 LuCl3 (25 mCi) was dissolved in 0.2 M sodium acetate buffer solution, and the reaction mixture was incubated at 90 °C for 1 hour to synthesize the structure having the structure of Formula 14 above. 177 Lu-DOTA-NCS-DMP. After cooling the reaction mixture at room temperature, 177 Lu-DOTA-NCS-DMP was separated and purified using a semi-preparative column.

[0137] 3-4: Preparation of 177Lu-DOTA-NCS-DMPY2

[0138]

[0139] 30 μg of DOTA-NCS-DMPY2 prepared in Examples 1-4 and 177 LuCl3 (23mCi) was dissolved in 0.2M sodium acetate buffer solution, and then the reaction mixture was incubated at 90°C for 1 hour to synthesize the structure having the structure of Formula 15 above. 177 Lu-DOTA-NCS-DMPY2. After cooling the reaction mixture at room temperature, 177 Lu-DOTA-NCS-DMPY2 was separated and purified using a semi-preparative column.

[0140] Experimental Example 1: In vivo anticancer activity test

[0141] The inventors administered the radioactive compound prepared in Examples 3-2 intravenously to a mouse model of melanoma and measured the changes in tumor volume and body weight over time.

[0142] Specifically, male BALB / c nu / nu mice (6 weeks old) were used and housed at the facility of Hwa Sun Hospital, College of Medicine, Chonnam National University. This research protocol was approved by the Institutional Animal Care and Use Committee (IACUC) of the College of Medicine, Chonnam National University.

[0143] 1×10 6 One B16F10 cell line (mouse melanoma cell line) was seeded into the right shoulder of male BALB / c nu / nu mice. The tumor volume increased to approximately 100 to 150 mm². 3 After stabilization, the material synthesized in Example 3-2 will be... 177Lu-DOTA-DMPY2 was administered intravenously to experimental animals at radiation doses of 90 MBq or 120 MBq, while the control group received only phosphate-buffered saline (PBS). Tumor volume (mm²) was recorded in the animals at 3-day intervals 21 days after administration. 3 The tumor volume was calculated using the following formula after measuring the width and length of the tumor tissue, along with the patient's weight:

[0144] Tumor volume (mm) 3 ) = (width) 2 ×(length)×0.5

[0145] The radioactive compound was administered only once at the start of treatment. After the experiment, the animals were euthanized, the tumor tissue was removed, and their weight was measured.

[0146] The results are as follows Figure 2 As shown, an application according to an embodiment of the present invention was performed. 177 In experimental animals, the size of tumors decreased significantly with Lu-DOTA-DMPY2, and in particular, the tumors showed almost no growth when the radiation dose was doubled.

[0147] In addition, the weight of the test animals is monitored to check for side effects of the experimental drug. Figure 3 The results shown are from measurements of changes in the body weight of experimental animals, applied according to an embodiment of the present invention. 177 Animals with Lu-DOTA-NCS-DMPY2 showed no significant difference in body weight compared to the control group.

[0148] Industrial applicability

[0149] As described above, the radioactive compounds according to embodiments of the present invention can be used very effectively to develop radiopharmaceuticals for the treatment of melanoma.

[0150] The above embodiments and experimental examples describe the present invention in more detail. However, the above embodiments and experimental examples are intended to describe the present invention more fully, and it will be apparent to those skilled in the art that the true scope of the present invention is not limited to the above embodiments and experimental examples. Therefore, the actual scope of protection of the present invention is as defined in the appended claims.

Claims

1. A radioactive compound or a pharmaceutically acceptable salt thereof, wherein the compound is selected from any one of the following compounds: 177 Lu-DOTA-DMPY2 177 Lu-DOTA-triazole-DMPY2 177 Lu-DOTA-triazole-PEG-DMPY2 177 Lu-DOTA-ADIBO-DMPY2 and, 2. A pharmaceutical composition for treating melanoma, comprising the radioactive compound of claim 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

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