Folate-drug conjugate, method for the preparation thereof, and pharmaceutical composition comprising the same

Abstract

The present invention relates to folic acid-drug conjugates, methods for preparing the same, and pharmaceutical compositions containing the same. The compound of the present invention, or its pharmaceutically acceptable salt, hydrate, solvate, or isomer, possesses excellent anticancer activity and an extended half-life in the body, thereby enabling sustained drug action and making it useful for the prevention or treatment of cancer. In particular, rather than possessing anticancer activity in itself, the compound of the present invention may exhibit pharmacological effects through a loaded drug released after reaching tumor tissue and a specific tumor microenvironment, or through active substances generated during metabolic processes. Furthermore, the compound of the present invention is a small molecule compound-drug conjugate to which an intelligent drug delivery system is applied, targeting folic acid receptor alpha, which is overexpressed in various cancer types, and the activated glucose uptake and metabolic characteristics unique to cancer cells. It exhibits tumor-specific drug delivery and cancer cell apoptosis effects while minimizing toxicity to normal cells. Therefore, based on its excellent anticancer effect and extended half-life in the body, the compound of the present invention is expected to be utilized as a promising therapeutic agent for the prevention or treatment of cancer and various related diseases.

Description

Technology Field

[0001] The present invention relates to a novel folic acid-drug conjugate that can be usefully used for the prevention or treatment of cancer, a method for preparing the same, and a pharmaceutical composition containing the same. Background Technology

[0003] Recently, smart drug delivery systems have been widely utilized in which a cytotoxic drug (payload) that cannot distinguish between normal and disease cells is conjugated to an antibody, and the drug is selectively delivered only to cells that overexpress the target antigen of the antibody. However, this method has limitations in that the drug hardly reaches disease cells that do not have a target antigen or express only a very small amount of it. To overcome these limitations, anticancer drugs in the form of Antibody-Drug Conjugates (ADCs) are gaining attention. These drugs enhance anticancer efficacy while minimizing the off-target toxicity of cytotoxic drugs, and also allow the use of cytotoxic drugs that were previously difficult to use as pharmaceuticals.

[0004] However, despite the development of various antibody-drug conjugates based on the inherent tumor specificity and selectivity of antibodies, factors such as the difficulty of penetrating into solid tumors, immunogenicity, and high manufacturing costs are still cited as limiting factors for clinical application. Accordingly, the inventors have invented a small molecule compound-drug conjugate, which is a new intelligent drug delivery platform that does not use antibodies. A novel folic acid-drug conjugate was developed having the ideal chemical structure required for a small molecule compound-drug conjugate, and furthermore, it was confirmed that the novel folic acid-drug conjugate can exhibit excellent tumor-specific drug delivery and anticancer effects. Prior art literature

[0006] (Republic of Korea Published Patent) No. 10-2024-0042109 The problem to be solved

[0007] The object of the present invention is to provide a compound represented by the following chemical formula 2, or a pharmaceutically acceptable salt thereof:

[0008] [Chemical Formula 2]

[0009]

[0010] In the above chemical formula 2,

[0011] A is amino C 1-10 Alkyl amido, amino-PEG 1~4 -C 1-3 Alkylamido, Amino-PEG 1~4 -C 1-2 Alkylamido C 1-10 Alkylamido, Lysylamido C 1-10 Alkyl amido, or amino-PEG 1~4 -C 1-2 Alkylamido lysylamido C 1-10 alkyl amido;

[0012] B is And, the above R1 is hydrogen or a carboxyl protector, and the above R2 is hydrogen or a hydroxyl protector, means a part connected to the above A or the above D;

[0013] LA is hydrogen, or and, above, R3 is hydrogen or a carboxyl protecting group, and n is 10 to 20, and represents a site connected to an amino acid residue in the above A;

[0014] D is is, means the part connected to the above B.

[0015] Another object of the present invention is (s1) a step of adding a compound represented by the following formula 10 to a compound represented by the following formula 9 to form an amide bond; and

[0016] (s2) A method for preparing a compound represented by the following chemical formula 2, comprising the step of deprotecting a hydroxyl protecting group or a carboxyl protecting group of the compound formed in step (s1):

[0017] [Chemical Formula 9]

[0018]

[0019] [Chemical Formula 10]

[0020]

[0021] [Chemical Formula 2]

[0022]

[0024] In the above chemical formula 9, R4 is a hydroxyl, amino C 1-10 Alkylcarboxylic acid, or amino-PEG 1~4 -C 1-3 It is an alkylcarboxylic acid, and

[0025] In the above chemical formula 10,

[0026] A is amino C 1-10 Alkyl amido, amino-PEG 1~4 -C 1-3 Alkylamido, Amino-PEG 1~4 -C 1-2 Alkylamido C 1-10 Alkylamido, Lysylamido C 1-10 Alkyl amido, or amino-PEG 1~4 -C 1-2 Alkylamido lysylamido C 1-10 It is an alkyl amido;

[0027] R1 is hydrogen, or a carboxyl protecting group;

[0028] R2 is hydrogen, or a hydroxyl protector;

[0029] LA is hydrogen, or And, the above R3 is hydrogen or a carboxyl protecting group, and n is 10 to 20, and represents the site connected to the amino acid residue in A above, and

[0030] In the above chemical formula 2,

[0031] A is amino C 1-10 Alkyl amido, amino-PEG 1~4 -C 1-3 Alkylamido, Amino-PEG 1~4 -C 1-2 Alkylamido C 1-10 Alkylamido, Lysylamido C 1-10 Alkyl amido, or amino-PEG 1~4 -C 1-2 Alkylamido lysylamido C 1-10 alkyl amido;

[0032] B is And, the above R1 is hydrogen or a carboxyl protector, and the above R2 is hydrogen or a hydroxyl protector, means a part connected to the above A or the above D;

[0033] LA is hydrogen, or And, the above R3 is hydrogen or a carboxyl protecting group, and n is 10 to 20, and represents the site connected to the amino acid residue in A above;

[0034] D is And, means the part connected to the above B.

[0035] Another object of the present invention is (s1) a step of adding a compound represented by the following formula 12 to a compound represented by the following formula 11 to form an amide bond, and

[0036] (s2) A method for preparing a compound represented by the following chemical formula 2, comprising the step of deprotecting a hydroxyl protecting group or a carboxyl protecting group of the compound formed in step (s1):

[0037] [Chemical Formula 11]

[0038]

[0039] [Chemical Formula 12]

[0040]

[0041] [Chemical Formula 2]

[0042]

[0043] In the above chemical formula 11, R1 is hydrogen or a carboxyl protecting group, and n is 10 to 20, and

[0044] In the above Chemical Formula 12, R1 is hydrogen or a carboxyl protecting group, R2 is hydrogen or a hydroxyl protecting group, and A is amino C 1-10 Alkyl amido, amino-PEG 1~4 -C 1-3 Alkylamido, Amino-PEG 1~4 -C 1-2 Alkylamido C 1-10 Alkylamido, Lysylamido C 1-10 Alkyl amido, or amino-PEG 1~4 -C 1-2 Alkylamido lysylamido C 1-10 alkyl amido,

[0045] In the above chemical formula 2,

[0046] A is amino C 1-10 Alkyl amido, amino-PEG 1~4 -C 1-3 Alkylamido, Amino-PEG 1~4 -C 1-2 Alkylamido C 1-10 Alkylamido, Lysylamido C 1-10 Alkyl amido, or amino-PEG 1~4 -C 1-2 Alkylamido lysylamido C 1-10 It is an alkyl amido;

[0047] B is And, the above R1 is hydrogen or a carboxyl protector, and the above R2 is hydrogen or a hydroxyl protector, and means a part connected to the above A or the above D;

[0048] LA is hydrogen, or And, the above R3 is hydrogen or a carboxyl protecting group, and n is 10 to 20, and represents the site connected to the amino acid residue in A above;

[0049] D is And, means the part connected to the above B.

[0050] Another object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of cancer, comprising the compound of the present invention or a pharmaceutically acceptable salt thereof as an active ingredient.

[0051] Another object of the present invention is to provide a drug delivery system for the prevention or treatment of cancer, comprising the compound of the present invention or a pharmaceutically acceptable salt thereof as an active ingredient.

[0053] However, the technical problems that the present invention aims to solve are not limited to those mentioned above, and other unmentioned problems will be clearly understood by those skilled in the art to which the present invention belongs from the description below. means of solving the problem

[0055] To achieve the above objectives, the present invention provides a compound represented by the following chemical formula 2, or a pharmaceutically acceptable salt thereof:

[0056] [Chemical Formula 2]

[0057]

[0058] In the above chemical formula 2,

[0059] A is amino C 1-10 Alkyl amido, amino-PEG 1~4 -C 1-3 Alkylamido, Amino-PEG 1~4 -C 1-2 Alkylamido C 1-10 Alkylamido, Lysylamido C 1-10 Alkyl amido, or amino-PEG 1~4 -C1-2 Alkylamido lysylamido C 1-10 alkyl amido;

[0060] B is And, the above R1 is hydrogen or a carboxyl protector, and the above R2 is hydrogen or a hydroxyl protector, means a part connected to the above A or the above D;

[0061] LA is hydrogen, or and, above, R3 is hydrogen or a carboxyl protecting group, and n is 10 to 20, and represents a site connected to an amino acid residue in the above A;

[0062] D is is, means the part connected to the above B.

[0063] In one embodiment of the present invention, n is 14 to 18; or the amino acid residue may be a lysine amino acid residue, but is not limited thereto.

[0064] In another embodiment of the present invention, the compound represented by Formula 2 may be characterized by one or more selected from the group consisting of the following, but is not limited thereto:

[0065] (a) A is amino C 1-10 Alkyl amido, amino-PEG 1~4 -C 1-2 Alkylamido C 1-10 Alkylamido, Lysylamido C 1-10 Alkyl amido, or amino-PEG 1~4 -C 1-2 Alkylamido lysylamido C 1-10 alkyl amido;

[0066] (b) B is And, means a part connected to the above A or the above D; and

[0067] (c) LA is hydrogen, or and, n is 14 to 18, and means the site connected to the amino acid residue in A above.

[0068] In another embodiment of the present invention, the compound represented by Formula 2 may be any one selected from the group consisting of the following, but is not limited thereto:

[0069] (1) (2R,3S,4S,5R)-6-(2-(2-(4-(4-(((2-amino-4-oxo-3,4-dihydropteridine-6-yl)methyl)amino)benzamido)-4-carboxybutanamido)acetamido)-4-(((((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[di]pyrano[3',4':6,7]yldolizino[1,2-b]quinoline-1-yl)carbamoyl)oxy)methyl)phenoxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylic acid;

[0070] (2) 1-(4-(((2-amino-4-oxo-3,4-dihydropteridine-6-yl)methyl)amino)phenyl)-8-((2-((2-((6-carboxy-3,4,5-trihydroxytetrahydro-2H-pyran-2-yl)oxy)-5-(((((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexa Hydro-1H,12H-benzo[di]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-1-yl)carbamoyl)oxy)methyl)phenyl)amino)-2-oxoethyl)carbamoyl)-1,6,14,23,32,37-hexaoxo-16,19,25,28-tetraoxo-2,7,13,22,31,36-hexaazatripentacontan-3,35,53-tricarboxylic acid;

[0071] (3) 19-(4-(((2-amino-4-oxo-3,4-dihydropteridine-6-yl)methyl)amino)benzamido)-1-((2-(((3R,4S,5S,6R)-6-carboxy-3,4,5-trihydroxytetrahydro-2H-pyran-2-yl)oxy)-5-(((((1S,9S)-9-ethyl-5-fluoro-9-hydroxy -4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[di]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-1-yl)carbamoyl)oxy)methyl)phenyl)amino)-1,4,16-trioxo-6,9,12-trioxa-3,15-diazaicosan-20-ican;

[0072] (4) 1-(4-(((2-amino-4-oxo-3,4-dihydropteridine-6-yl)methyl)amino)phenyl)-17-((2-((2-(((3R,4S,5S,6R)-6-carboxy-3,4,5-trihydroxytetrahydro-2H-pyran-2-yl)oxy)-5-(((((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15- Hexahydro-1H,12H-benzo[di]pyrano[3',4':6,7]indoleizino[1,2-b]quinoline-1-yl)carbamoyl)oxy)methyl)phenyl)amino)-2-oxoethyl)carbamoyl)-1,6,15,23,32,41,46-heptaoxo-10,13,25,28,34,37-hexaoxo-2,7,16,22,31,40,45-heptazadohexacontan-3,44,62-tricarboxylic acid;

[0073] (5) 20-(4-(((2-amino-4-oxo-3,4-dihydropteridine-6-yl)methyl)amino)benzamido)-1-((2-(((2R,3R,4S,5S,6R)-6-carboxy-3,4,5-trihydroxytetrahydro-2H-pyran-2-yl)oxy)-5-(((((1S,9S)-9-ethyl-5-fluoro-9-hydroxy -4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[di]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-1-yl)carbamoyl)oxy)methyl)phenyl)amino)-1,5,17-trioxo-7,10,13-trioxa-4,16-diazahenicosan-21-ican; and

[0074] (6) (2R,3S,4S,5R)-6-(2-(3-(4-(4-(4-(((2-amino-4-oxo-3,4-dihydropteridine-6-yl)methyl)amino)benzamido)-4-carboxybutanamido)butanamido)propaneamido)-4-(((((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[di]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-1-yl)carbamoyl)oxy)methyl)phenoxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylic acid.

[0075] In another embodiment of the present invention, the compound represented by Formula 2 may be any one selected from the group consisting of Formulas 3 to 8 below, but is not limited thereto.

[0076] [Chemical Formula 3]

[0077]

[0078] [Chemical Formula 4]

[0079]

[0080] [Chemical Formula 5]

[0081]

[0082] [Chemical Formula 6]

[0083]

[0084] [Chemical Formula 7]

[0085]

[0086] [Chemical Formula 8]

[0087]

[0088] The present invention comprises (s1) a step of adding a compound represented by the following chemical formula 10 to a compound represented by the following chemical formula 9 to form an amide bond; and

[0089] (s2) A method for preparing a compound represented by the following chemical formula 2 is provided, comprising the step of deprotecting a hydroxyl protecting group or a carboxyl protecting group of the compound formed in step (s1):

[0090] [Chemical Formula 9]

[0091]

[0092] [Chemical Formula 10]

[0093]

[0094] [Chemical Formula 2]

[0095]

[0097] In the above chemical formula 9, R4 is a hydroxyl, amino C 1-10 Alkylcarboxylic acid, or amino-PEG 1~4 -C 1-3 It is an alkylcarboxylic acid, and

[0098] In the above chemical formula 10,

[0099] A is amino C 1-10 Alkyl amido, amino-PEG 1~4 -C 1-3 Alkylamido, Amino-PEG 1~4 -C 1-2 Alkylamido C 1-10 Alkylamido, Lysylamido C 1-10 Alkyl amido, or amino-PEG 1~4 -C 1-2 Alkylamido lysylamido C 1-10 It is an alkyl amido;

[0100] R1 is hydrogen, or a carboxyl protecting group;

[0101] R2 is hydrogen, or a hydroxyl protector;

[0102] LA is hydrogen, or And, the above R3 is hydrogen or a carboxyl protecting group, and n is 10 to 20, and represents the site connected to the amino acid residue in A above, and

[0103] In the above chemical formula 2,

[0104] A is amino C 1-10 Alkyl amido, amino-PEG 1~4 -C 1-3 Alkylamido, Amino-PEG 1~4 -C 1-2 Alkylamido C 1-10 Alkylamido, Lysylamido C 1-10 Alkyl amido, or amino-PEG 1~4 -C 1-2 Alkylamido lysylamido C 1-10 alkyl amido;

[0105] B is And, the above R1 is hydrogen or a carboxyl protector, and the above R2 is hydrogen or a hydroxyl protector, means a part connected to the above A or the above D;

[0106] LA is hydrogen, or And, the above R3 is hydrogen or a carboxyl protecting group, and n is 10 to 20, and represents the site connected to the amino acid residue in A above;

[0107] D is And, means the part connected to the above B.

[0108] In one embodiment of the present invention, the compounds represented by Formula 9, Formula 10, and Formula 2 may be characterized by one or more selected from the group consisting of the following, but are not limited thereto:

[0109] (a) In the above chemical formula 9, R4 is a hydroxyl, amino C 1-4 Alkylcarboxylic acid, or amino-PEG 1~4 -C 1-3 alkylcarboxylic acid,

[0110] (b) In the above chemical formula 10, A is amino C 1-10 Alkyl amido, amino-PEG 1~4 -C1-2 Alkylamido C 1-10 Alkylamido, Lysylamido C 1-10 Alkyl amido, or amino-PEG 1~4 -C 1-2 Alkylamido lysylamido C 1-10 It is an alkyl amido, R1 is hydrogen or a carboxyl protecting group, R2 is hydrogen or a hydroxyl protecting group, and LA is hydrogen or and, n is 14 to 18, and means the site connected to the amino acid residue in A above, and

[0111] (c) In the above chemical formula 2,

[0112] A is amino C 1-10 Alkyl amido, amino-PEG 1~4 -C 1-2 Alkylamido C 1-10 Alkylamido, Lysylamido C 1-10 Alkyl amido, or amino-PEG 1~4 -C 1-2 Alkylamido lysylamido C 1-10 It is an alkyl amido;

[0113] B is And, means a part connected to the above A or the above D; and

[0114] LA is hydrogen, or and, n is 14 to 18, and means the site connected to the lysine amino acid residue in A above.

[0115] The present invention comprises (s1) a step of adding a compound represented by the following chemical formula 12 to a compound represented by the following chemical formula 11 to form an amide bond, and

[0116] (s2) A method for preparing a compound represented by the following chemical formula 2 is provided, comprising the step of deprotecting a hydroxyl protecting group or a carboxyl protecting group of the compound formed in step (s1):

[0117] [Chemical Formula 11]

[0118]

[0119] [Chemical Formula 12]

[0120]

[0121] [Chemical Formula 2]

[0122]

[0123] In the above chemical formula 11, R1 is hydrogen or a carboxyl protecting group, and n is 10 to 20, and

[0124] In the above Chemical Formula 12, R1 is hydrogen or a carboxyl protecting group, R2 is hydrogen or a hydroxyl protecting group, and A is amino C 1-10 Alkyl amido, amino-PEG 1~4 -C 1-3 Alkylamido, Amino-PEG 1~4 -C 1-2 Alkylamido C 1-10 Alkylamido, Lysylamido C 1-10 Alkyl amido, or amino-PEG 1~4 -C 1-2 Alkylamido lysylamido C 1-10 alkyl amido,

[0125] In the above chemical formula 2,

[0126] A is amino C 1-10 Alkyl amido, amino-PEG 1~4 -C 1-3 Alkylamido, Amino-PEG 1~4 -C 1-2 Alkylamido C 1-10 Alkylamido, Lysylamido C 1-10 Alkyl amido, or amino-PEG 1~4 -C 1-2 Alkylamido lysylamido C 1-10 It is an alkyl amido;

[0127] B is And, the above R1 is hydrogen or a carboxyl protector, and the above R2 is hydrogen or a hydroxyl protector, and means a part connected to the above A or the above D;

[0128] LA is hydrogen, or And, the above R3 is hydrogen or a carboxyl protecting group, and n is 10 to 20, and represents the site connected to the amino acid residue in A above;

[0129] D is And, means the part connected to the above B.

[0130] In one embodiment of the present invention, the compound represented by Formula 11, Formula 12, and Formula 2 may be characterized by one or more selected from the group consisting of the following, but is not limited thereto:

[0131] (a) In the above chemical formula 11, R1 is hydrogen or a carboxyl protecting group, and n is 14 to 18,

[0132] (b) In the above Chemical Formula 12, R1 is hydrogen or a carboxyl protecting group, R2 is hydrogen or a hydroxyl protecting group, and A is amino C 1-3 alkyl amido, and

[0133] (c) In the above chemical formula 2,

[0134] A is amino C 1-10 Alkyl amido, amino-PEG 1~4 -C 1-2 Alkylamido C 1-10 Alkylamido, Lysylamido C 1-10 Alkyl amido, or amino-PEG 1~4 -C 1-2 Alkylamido lysylamido C 1-10 It is an alkyl amido;

[0135] B is And, means a part connected to the above A or the above D; and

[0136] LA is hydrogen, or and, n is 14 to 18, and means the site connected to the amino acid residue in A above.

[0137] In another embodiment of the present invention, the compound represented by the formula 11 is,

[0138] (a) a step of synthesizing a compound represented by the following chemical formula 14 by reacting a compound represented by the following chemical formula 13 with a solid phase support into which 2-chlorotrityl chloride has been introduced,

[0139] (b) a step of adding a fluorenylmethoxycarbonyl protecting group (9-Fluorenylmethyloxycarbonyl, Fmoc)-L-glutamic acid 5-tert-butyl ester to the compound formed in step (a) above to form an amide bond,

[0140] (c) a step of adding N-(trifluoroacetyl)pteroic acid to the compound formed in step (b) above to form an amide bond, and

[0141] (d) It may be manufactured by a manufacturing method comprising one or more steps selected from the group consisting of removing a solid support from the compound formed in step (c), but is not limited thereto:

[0142] [Chemical Formula 13]

[0144]

[0145] [Chemical Formula 14]

[0146]

[0147] In the above chemical formulas 13 and 14, respectively, R1 is hydrogen or a carboxyl protecting group, and n is 14 to 18.

[0148] In another embodiment of the present invention, steps (b) and (c) may each be performed independently after deprotecting the fluorenylmethoxycarbonyl protecting group (9-Fluorenylmethyloxycarbonyl, Fmoc), but are not limited thereto.

[0149] The present invention provides a pharmaceutical composition for the prevention or treatment of cancer, comprising the compound of the present invention or a pharmaceutically acceptable salt thereof as an active ingredient.

[0150] In one embodiment of the present invention, the cancer may be one or more selected from the group consisting of ovarian cancer, breast cancer, kidney cancer, lung cancer, colorectal cancer, and brain cancer, but is not limited thereto.

[0151] The present invention provides a drug delivery system for the prevention or treatment of cancer, comprising the compound of the present invention or a pharmaceutically acceptable salt thereof as an active ingredient.

[0152] In one embodiment of the present invention, the cancer may be one or more selected from the group consisting of ovarian cancer, breast cancer, kidney cancer, lung cancer, colorectal cancer, and brain cancer, but is not limited thereto.

[0154] In addition, the present invention provides a method for preventing, improving, or treating cancer, comprising the step of administering a pharmaceutically effective amount of a composition containing the compound of the present invention or a pharmaceutically acceptable salt thereof as an active ingredient to an individual in need thereof.

[0155] In addition, the present invention provides a use for cancer prevention, improvement, or treatment of a composition comprising the compound of the present invention or a pharmaceutically acceptable salt thereof as an active ingredient.

[0156] In addition, the present invention provides a use for preparing a cancer prevention, improvement, or treatment agent of a composition comprising the compound of the present invention or a pharmaceutically acceptable salt thereof as an active ingredient. Effects of the invention

[0158] The compound of the present invention, or its pharmaceutically acceptable salt, hydrate, solvate, or isomer, possesses excellent anticancer activity and has an extended half-life in the body; by enabling sustained drug action, it can be usefully employed in the prevention or treatment of cancer. In particular, rather than possessing anticancer activity in itself, the compound of the present invention may exhibit pharmacological effects through a loaded drug released after reaching tumor tissue and a specific tumor microenvironment, or through active substances generated during metabolic processes. Furthermore, the compound of the present invention is a small molecule compound-drug conjugate to which an intelligent drug delivery system is applied, targeting folate receptor alpha, which is overexpressed in various cancer types, and the activated glucose uptake and metabolic characteristics unique to cancer cells. It exhibits tumor-specific drug delivery and cancer cell apoptosis effects while minimizing toxicity to normal cells. Therefore, based on its excellent anticancer effect and extended half-life in the body, the compound of the present invention is expected to be utilized as a promising therapeutic agent for the prevention or treatment of cancer and various related diseases. Brief explanation of the drawing

[0160] Figure 1 shows a compound represented by Chemical Formula 2 of the present invention. Specific details for implementing the invention

[0161] The present invention provides a compound represented by the following chemical formula 2, or a pharmaceutically acceptable salt thereof:

[0162] [Chemical Formula 2]

[0163]

[0164] In the above chemical formula 2,

[0165] A is amino C 1-10 Alkyl amido, amino-PEG 1~4 -C 1-3 Alkylamido, Amino-PEG 1~4 -C1-2 Alkylamido C 1-10 Alkylamido, Lysylamido C 1-10 Alkyl amido, or amino-PEG 1~4 -C 1-2 Alkylamido lysylamido C 1-10 alkyl amido;

[0166] B is And, the above R1 is hydrogen or a carboxyl protector, and the above R2 is hydrogen or a hydroxyl protector, means a part connected to the above A or the above D;

[0167] LA is hydrogen, or and, above, R3 is hydrogen or a carboxyl protecting group, and n is 10 to 20, and represents a site connected to an amino acid residue in the above A;

[0168] D is is, means a region connected to the above B, and specifically, may mean a region connected to a carboxyl on the above B.

[0169] In the present invention, the “hydroxyl protecting group” includes all groups that can be used as a protecting group for a conventional hydroxyl group, and specifically, may be an acyl group, an alkyloxycarbonyl group, an aralkyloxycarbonyl group, a heterocyclic oxycarbonyl group, an alkyl group, an alkenyl group, an aralkyl group, an oxygen-containing heterocyclic group, a sulfur-containing heterocyclic group, an alkoxyalkyl group, an aralkyloxyalkyl group, an alkylsulfonyl group, an arylsulfonyl group, and a substituted silyl group, but is not limited thereto.

[0170] In addition, in one embodiment of the present invention, the hydroxyl protector is an acyl group, C 1-12 Alkyloxycarbonyl group, Ar-C 1-6 Alkyl oxycarbonyl group, heterocyclic oxycarbonyl group, C 1-12 Alkyl group, C 2-12 alkenyl group, Ar-C 1-6 Alkyl group, oxygen-containing polycyclic group, sulfur-containing polycyclic group, C1-6 Alkyloxy C 1-6 Alkyl group, ArC 1-6 Alkyloxy C 1-6 Alkyl group, C 1-12 The alkylsulfonyl group, arylsulfonyl group, or substituted silyl group is, wherein the aryl group among the hydroxyl protecting groups may be a phenyl, naphthyl, indanyl, or indenyl group, and specifically, in one embodiment of the present invention, the hydroxyl protecting group is an acyl group, C 1-12 alkyl group, or C 1-6 Alkyloxy C 1-6 It may be an alkyl group, but is not limited thereto.

[0171] In the present invention, the “carboxyl protecting group” includes all groups that can be used as a protecting group for a conventional carboxyl group, and specifically, examples include alkyl groups, alkenyl groups, aryl groups, aralkyl groups, acylalkyl groups, arylthioalkyl groups, arylsulfonylalkyl groups, oxygen-containing heterocyclic groups, alkylsilylalkyl groups, acyloxyalkyl groups, nitrogen-containing heterocyclic alkyl groups, cycloalkyl groups, alkoxyalkyl groups, aralkyloxyalkyl groups, alkylthioalkyl groups, and substituted silyl groups.

[0172] In addition, in one embodiment of the present invention, the carboxyl protector is C 1-12 Alkyl group, C 2-12 Alkenyl group, aryl group, Ar-C 1-6 Alkyl group, acylalkyl group, arylthioalkyl group, arylsulfonylalkyl group, oxygen-containing heterocyclic group, alkylsilylalkyl group, acyloxyalkyl group, nitrogen-containing heterocyclic alkyl group, C 3-8 Cycloalkyl group, C 1-6 Alkyloxy C 1-6 Alkyl group, ArC 1-6 Alkyloxy C 1-6 Alkyl group, C 1-6 Alkylthio C 1-6 The alkyl group or substituted silyl group is, wherein the aryl group among the carboxyl protecting groups may be a phenyl, naphthyl, indanyl, or indenyl group, and specifically, in one embodiment of the present invention, the carboxyl protecting group is C 1-12 Alkyl group, acylalkyl group, or C 1-6 Alkyloxy C1-6 It may be an alkyl group, but is not limited thereto.

[0173] In one embodiment of the present invention, n may be 10 to 18, 10 to 17, 10 to 16, 10 to 15, 10 to 14, 12 to 18, 12 to 17, 12 to 16, 12 to 15, 12 to 14, 13 to 18, 13 to 17, 13 to 16, 13 to 15, 13 to 14, 14 to 18, 14 to 17, 14 to 16, 14 to 15, 15 to 18, 15 to 17, 15 to 16, 16 to 18, 16 to 17, 14, 15, 16, 17, or 18, but is not limited to.

[0174] In one embodiment of the present invention, the amino acid residue may be a lysine amino acid residue, but is not limited thereto.

[0175] In one embodiment of the present invention, the compound represented by Formula 2 may be a compound represented by Formula 1 below when LA is hydrogen, but is not limited thereto:

[0176] [Chemical Formula 1]

[0177] .

[0178] In the present invention, “Folate receptor α (FRα)” is a target for the development of anticancer drugs that regulate folate metabolism. Since folate receptor α is overexpressed in a number of solid tumors, including ovarian cancer, lung cancer, and breast cancer, various approaches for the diagnosis and treatment of these cancers are possible. Accordingly, the inventors have invented a small molecule compound-drug conjugate that selectively delivers drugs to tumor cells overexpressing folate receptor α by utilizing the principle that folic acid derivatives strongly bind to folate receptor α. Since the drug hardly reaches normal cells that lack folate receptor α or have minimal expression, this small molecule compound-drug conjugate of the present invention can exhibit the advantage of reducing off-target toxicity of cytotoxic drugs and enhancing anticancer efficacy.

[0179] In the present invention, “cancer cells” undergo more active glucose absorption and metabolism than normal cells for rapid proliferation, and in fact, in cancer cells of many types of cancer, glucose transporters, glucose receptors, as well as various glucose hydrolyzing enzymes such as glycosids are overexpressed. For example, high expression of beta-glucuronidase is observed in ovarian cancer, lung cancer, breast cancer, pancreatic cancer, colorectal cancer, etc. The inventors have invented a glucose-based drug delivery system utilizing a glycosylated prodrug for cancer treatment by taking advantage of these characteristics of cancer cells. Specifically, the glycosylated prodrug contained in the compound of the present invention is designed to accumulate specifically in cancer cells and to release the drug through glucose-metabolizing enzymes overexpressed within the cancer cells, thereby enhancing the anticancer effect and minimizing toxicity to normal cells. Furthermore, the glucose portion contained in the compound of the present invention has high water solubility, low toxicity, and high biocompatibility, which can exhibit the advantage of improving the pharmacokinetics of the drug.

[0180] In one embodiment of the present invention, the compound may be a low-molecular-weight compound-drug conjugate, but is not limited thereto.

[0181] In the present invention, the “small molecule compound-drug conjugate” may be composed of four parts: a targeting ligand, a spacer linker, a cleavable linker, and a payload / therapeutic cargo, drug, but is not limited thereto.

[0182] In the present invention, the “target ligand” is a ligand that specifically recognizes a target overexpressed in a disease and selectively binds to it to act, and includes receptor inhibitors / agonists and other regulatory substances, and the target ligand may be a small molecule compound (small molecule or peptide), and includes all that can specifically recognize a target overexpressed in a disease within the scope of the ordinary level of technology.

[0183] In the present invention, the “spacer linker” may be water-soluble in order to minimize non-selective passive permeation without interfering with the binding of the target ligand to the disease target, together with the cleavage linker, and may play a role in improving the pharmacokinetics of the small molecule compound-drug conjugate. Additionally, in one embodiment of the present invention, the spacer linker may be denoted by A.

[0184] In the present invention, the “cleavable linker” is a region responsible for releasing the loaded drug after the small molecule compound-drug conjugate binds to a disease target, and may allow the drug to be released through endogenous enzyme systems or substances that are activated or more abundant in the disease target and environment. The cleavable linker may include a sugar, but is not limited thereto. Additionally, in one embodiment of the present invention, the cleavable linker may be denoted by B.

[0185] In the present invention, the “payload / therapeutic cargo, drug” is a drug that performs the role of being finally released to exert a therapeutic effect after being delivered to a disease target, and potent cytotoxic drugs are primarily used for the purpose of cancer treatment. The cytotoxic drug may be a broad concept including substances capable of killing cancer cells, such as DNA damaging agents, microtubule inhibitors, and topoisomerase I inhibitors. Additionally, in one embodiment of the present invention, the payload drug may be denoted by D.

[0186] In the present invention, the “long-acting site” refers to a portion intended to improve the drug’s persistence by extending the half-life of the compound of the present invention. In the treatment of diseases including cancer, maintaining an effective drug concentration within the target tissue for a certain period of time is a critical factor in the manifestation of drug efficacy; accordingly, drug delivery systems are designed to simultaneously consider not only specificity and selectivity for the target tissue but also persistence within the body. Accordingly, the inventors of the present invention applied a strategy of introducing a long-acting site into a small molecule compound-drug conjugate. In particular, for small molecule-drug conjugate drugs with a short blood half-life, a fatty acid derivative capable of strongly binding to albumin in the body was introduced to extend the blood half-life, thereby improving the drug’s persistence. Furthermore, in one embodiment of the present invention, the long-acting site may be denoted as LA.

[0187] In the present invention, the spacer linker, cleavable linker, long-acting site, and payload / therapeutic cargo, drug may comprise, but are not limited to, the compounds described above and pharmaceutically acceptable salts thereof.

[0188] In the present invention, the amino-PEG 1~4 -C 1-2 Alkylamido is , , , , or and, here may be a site connected to a target ligand, a space linker (A), or a cutting linker (B), but is not limited thereto.

[0189] In one embodiment of the present invention, the compound represented by Formula 2 may be characterized by one or more selected from the group consisting of the following, but is not limited thereto:

[0190] (a) A is amino C 1-10 Alkyl amido, amino-PEG 1~4 -C 1-2 Alkylamido C 1-10 Alkylamido, Lysylamido C 1-10 Alkyl amido, or amino-PEG 1~4 -C 1-2 Alkylamido lysylamido C 1-10 alkyl amido;

[0191] (b) B is And, means a part connected to the above A or the above D; and

[0192] (c) LA is hydrogen, or

[0193] and, n is 14 to 18, and means the site connected to the amino acid residue in A above.

[0194] In one embodiment of the present invention, A of the compound represented by Chemical Formula 2 is amino C 1-3 Alkyl amido, amino-PEG 1-3 -C 1-2 Alkylamido C 1-3 Alkylamido, Lysylamido C 1-3 Alkyl amido, or amino-PEG 1-3 -C 1-2 Alkylamido lysylamido C 1-3 It may be an alkyl amido, but is not limited thereto.

[0195] In one embodiment of the present invention, A of the compound represented by Chemical Formula 2 is amino C 1-2 Alkyl amido, amino-PEG3-C1alkylamido C 1-2It may be an alkylamido, a lysylamido C1-alkylamido, or an amino-PEG2-C1-alkylamido lysylamido C1-alkylamido, but is not limited thereto.

[0196] In one embodiment of the present invention, compound A represented by the formula 2 may be amino C1-alkylamido, amino C2-alkylamido, amino-PEG3-C1-alkylamido C1-alkylamido, amino-PEG3-C1-alkylamido C2-alkylamido, lysylamido C1-alkylamido, or amino-PEG2-C1-alkylamido lysylamido C1-alkylamido, but is not limited thereto.

[0197] In one embodiment of the present invention, the compound represented by Formula 2 may be any one selected from the group consisting of the following, but is not limited thereto:

[0198] (1) (2R,3S,4S,5R)-6-(2-(2-(4-(4-(((2-amino-4-oxo-3,4-dihydropteridine-6-yl)methyl)amino)benzamido)-4-carboxybutanamido)acetamido)-4-(((((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[di]pyrano[3',4':6,7]yldolizino[1,2-b]quinoline-1-yl)carbamoyl)oxy)methyl)phenoxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylic acid;

[0199] (2) 1-(4-(((2-amino-4-oxo-3,4-dihydropteridine-6-yl)methyl)amino)phenyl)-8-((2-((2-((6-carboxy-3,4,5-trihydroxytetrahydro-2H-pyran-2-yl)oxy)-5-(((((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexa Hydro-1H,12H-benzo[di]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-1-yl)carbamoyl)oxy)methyl)phenyl)amino)-2-oxoethyl)carbamoyl)-1,6,14,23,32,37-hexaoxo-16,19,25,28-tetraoxo-2,7,13,22,31,36-hexaazatripentacontan-3,35,53-tricarboxylic acid;

[0200] (3) 19-(4-(((2-amino-4-oxo-3,4-dihydropteridine-6-yl)methyl)amino)benzamido)-1-((2-(((3R,4S,5S,6R)-6-carboxy-3,4,5-trihydroxytetrahydro-2H-pyran-2-yl)oxy)-5-(((((1S,9S)-9-ethyl-5-fluoro-9-hydroxy -4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[di]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-1-yl)carbamoyl)oxy)methyl)phenyl)amino)-1,4,16-trioxo-6,9,12-trioxa-3,15-diazaicosan-20-ican;

[0201] (4) 1-(4-(((2-amino-4-oxo-3,4-dihydropteridine-6-yl)methyl)amino)phenyl)-17-((2-((2-(((3R,4S,5S,6R)-6-carboxy-3,4,5-trihydroxytetrahydro-2H-pyran-2-yl)oxy)-5-(((((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15- Hexahydro-1H,12H-benzo[di]pyrano[3',4':6,7]indoleizino[1,2-b]quinoline-1-yl)carbamoyl)oxy)methyl)phenyl)amino)-2-oxoethyl)carbamoyl)-1,6,15,23,32,41,46-heptaoxo-10,13,25,28,34,37-hexaoxo-2,7,16,22,31,40,45-heptazadohexacontan-3,44,62-tricarboxylic acid;

[0202] (5) 20-(4-(((2-amino-4-oxo-3,4-dihydropteridine-6-yl)methyl)amino)benzamido)-1-((2-(((2R,3R,4S,5S,6R)-6-carboxy-3,4,5-trihydroxytetrahydro-2H-pyran-2-yl)oxy)-5-(((((1S,9S)-9-ethyl-5-fluoro-9-hydroxy -4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[di]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-1-yl)carbamoyl)oxy)methyl)phenyl)amino)-1,5,17-trioxo-7,10,13-trioxa-4,16-diazahenicosan-21-ican; and

[0203] (6) (2R,3S,4S,5R)-6-(2-(3-(4-(4-(4-(((2-amino-4-oxo-3,4-dihydropteridine-6-yl)methyl)amino)benzamido)-4-carboxybutanamido)butanamido)propaneamido)-4-(((((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[di]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-1-yl)carbamoyl)oxy)methyl)phenoxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylic acid.

[0204] In one embodiment of the present invention, the compound represented by Formula 2 may be any one selected from the group consisting of Formulas 3 to 8 below, but is not limited thereto.

[0205] [Chemical Formula 3]

[0206]

[0207] [Chemical Formula 4]

[0208]

[0209] [Chemical Formula 5]

[0210]

[0211] [Chemical Formula 6]

[0212]

[0213] [Chemical Formula 7]

[0214]

[0215] [Chemical Formula 8]

[0216]

[0217] The present invention comprises (s1) a step of adding a compound represented by the following chemical formula 10 to a compound represented by the following chemical formula 9 to form an amide bond; and

[0218] (s2) A method for preparing a compound represented by the following chemical formula 2 is provided, comprising the step of deprotecting a hydroxyl protecting group or a carboxyl protecting group of the compound formed in step (s1):

[0219] [Chemical Formula 9]

[0220]

[0221] [Chemical Formula 10]

[0222]

[0223] [Chemical Formula 2]

[0224]

[0226] In the above chemical formula 9, R4 is a hydroxyl, amino C 1-10 Alkylcarboxylic acid, or amino-PEG 1~4 -C 1-3 It is an alkylcarboxylic acid, and

[0227] In the above chemical formula 10,

[0228] A is amino C 1-10 Alkyl amido, amino-PEG 1~4 -C 1-3 Alkylamido, Amino-PEG 1~4 -C 1-2 Alkylamido C 1-10 Alkylamido, Lysylamido C 1-10 Alkyl amido, or amino-PEG 1~4 -C 1-2 Alkylamido lysylamido C 1-10 It is an alkyl amido;

[0229] R1 is hydrogen, or a carboxyl protecting group;

[0230] R2 is hydrogen, or a hydroxyl protector;

[0231] LA is hydrogen, or And, the above R3 is hydrogen or a carboxyl protecting group, and n is 10 to 20, and represents the site connected to the amino acid residue in A above, and

[0232] In the above chemical formula 2,

[0233] A is amino C 1-10 Alkyl amido, amino-PEG 1~4 -C 1-3 Alkylamido, Amino-PEG 1~4 -C 1-2 Alkylamido C 1-10 Alkylamido, Lysylamido C 1-10 Alkyl amido, or amino-PEG 1~4 -C 1-2 Alkylamido lysylamido C 1-10 alkyl amido;

[0234] B is And, the above R1 is hydrogen or a carboxyl protector, and the above R2 is hydrogen or a hydroxyl protector, means a part connected to the above A or the above D;

[0235] LA is hydrogen, or And, the above R3 is hydrogen or a carboxyl protecting group, and n is 10 to 20, and represents the site connected to the amino acid residue in A above;

[0236] D is And, means the part connected to the above B.

[0237] In one embodiment of the present invention, R2 is hydrogen or a hydroxyl protector, wherein may be a part connected to A or D, but is not limited thereto.

[0238] In one embodiment of the present invention, the compounds represented by Formula 9, Formula 10, and Formula 2 may be characterized by one or more selected from the group consisting of the following, but are not limited thereto:

[0239] (a) In the above chemical formula 9, R4 is a hydroxyl, amino C 1-4 Alkylcarboxylic acid, or amino-PEG 1~4 -C 1-3 alkylcarboxylic acid,

[0240] (b) In the above chemical formula 10, A is amino C 1-10 Alkyl amido, amino-PEG 1~4 -C 1-2 Alkylamido C 1-10 Alkylamido, Lysylamido C 1-10 Alkyl amido, or amino-PEG 1~4 -C 1-2 Alkylamido lysylamido C 1-10It is an alkyl amido, R1 is hydrogen or a carboxyl protecting group, R2 is hydrogen or a hydroxyl protecting group, and LA is hydrogen or and, n is 14 to 18, and means the site connected to the amino acid residue in A above, and

[0241] (c) In the above chemical formula 2,

[0242] A is amino C 1-10 Alkyl amido, amino-PEG 1~4 -C 1-2 Alkylamido C 1-10 Alkylamido, Lysylamido C 1-10 Alkyl amido, or amino-PEG 1~4 -C 1-2 Alkylamido lysylamido C 1-10 It is an alkyl amido;

[0243] B is And, means a part connected to the above A or the above D; and

[0244] LA is hydrogen, or and, n is 14 to 18, and means the site connected to the lysine amino acid residue in A above.

[0245] In the present invention, in the above formula 9, R4 may be a hydroxyl, an amino C3 alkylcarboxylic acid, an amino-PEG3-C1 alkylcarboxylic acid, or an amino-PEG2-C1 alkylcarboxylic acid, but is not limited thereto.

[0247] In the present invention, the step (s1) may be performed in the presence of one or more selected from the group consisting of N,N'-Dicyclohexylcarbodiimide, N-Hydroxysuccinimide, pyridine, dimethyl sulfoxide (DMSO), and dichloromethyl (DCM), but is not limited thereto.

[0248] In the present invention, the step (s2) may be performed in the presence of one or more selected from the group consisting of methanol (MeOH), lithium hydroxide (LiOH), and hydrochloric acid (HCl), but is not limited thereto.

[0249] In one embodiment of the present invention, a method for preparing a compound represented by Formula 3 or a pharmaceutically acceptable salt thereof may be as shown in Reaction Scheme 1 below, but is not limited thereto:

[0250] [Reaction Equation 1]

[0251]

[0252] In the present invention, A, R1, and R2 in Reaction Scheme 1 may be as described above, but are not limited thereto. Additionally, Reaction Scheme 1 may be formed by removing hydroxyl and carboxyl protecting groups after a conventionally used amide bond reaction.

[0253] In one embodiment of the present invention, a method for preparing a compound represented by Formula 4 or a pharmaceutically acceptable salt thereof may be as shown in Reaction Scheme 2 below, but is not limited thereto:

[0254] [Reaction Equation 2]

[0255]

[0256] In the present invention, A, LA, R1, and R2 in Reaction Scheme 2 may be as described above, but are not limited thereto. Additionally, Reaction Scheme 2 may be formed by removing hydroxyl and carboxyl protecting groups after a conventionally used amide bond reaction.

[0257] In the present invention, the above reaction formula 1 or reaction formula 2 may replace folic acid with a compound represented by chemical formula 9, and reaction formula 1 or reaction formula 2 may be a method for preparing a compound represented by chemical formulas 5 to 8 or a pharmaceutically acceptable salt thereof, but is not limited thereto.

[0258] The present invention comprises (s1) a step of adding a compound represented by the following chemical formula 12 to a compound represented by the following chemical formula 11 to form an amide bond, and

[0259] (s2) A method for preparing a compound represented by the following chemical formula 2 is provided, comprising the step of deprotecting a hydroxyl protecting group or a carboxyl protecting group of the compound formed in step (s1):

[0260] [Chemical Formula 11]

[0261]

[0262] [Chemical Formula 12]

[0263]

[0264] [Chemical Formula 2]

[0265]

[0266] In the above chemical formula 11, R1 is hydrogen or a carboxyl protecting group, and n is 10 to 20, and

[0267] In the above Chemical Formula 12, R1 is hydrogen or a carboxyl protecting group, R2 is hydrogen or a hydroxyl protecting group, and A is amino C 1-10 Alkyl amido, amino-PEG 1~4 -C 1-3Alkylamido, Amino-PEG 1~4 -C 1-2 Alkylamido C 1-10 Alkylamido, Lysylamido C 1-10 Alkyl amido, or amino-PEG 1~4 -C 1-2 Alkylamido lysylamido C 1-10 alkyl amido,

[0268] In the above chemical formula 2,

[0269] A is amino C 1-10 Alkyl amido, amino-PEG 1~4 -C 1-3 Alkylamido, Amino-PEG 1~4 -C 1-2 Alkylamido C 1-10 Alkylamido, Lysylamido C 1-10 Alkyl amido, or amino-PEG 1~4 -C 1-2 Alkylamido lysylamido C 1-10 It is an alkyl amido;

[0270] B is And, the above R1 is hydrogen or a carboxyl protector, and the above R2 is hydrogen or a hydroxyl protector, and means a part connected to the above A or the above D;

[0271] LA is hydrogen, or And, the above R3 is hydrogen or a carboxyl protecting group, and n is 10 to 20, and represents the site connected to the amino acid residue in A above;

[0272] D is And, means the part connected to the above B.

[0273] In one embodiment of the present invention, the compound represented by Formula 11, Formula 12, and Formula 2 may be characterized by one or more selected from the group consisting of the following, but is not limited thereto:

[0274] (a) In the above chemical formula 11, R1 is hydrogen or a carboxyl protecting group, and n is 14 to 18,

[0275] (b) In the above Chemical Formula 12, R1 is hydrogen or a carboxyl protecting group, R2 is hydrogen or a hydroxyl protecting group, and A is amino C 1-3 alkyl amido, and

[0276] (c) In the above chemical formula 2,

[0277] A is amino C 1-10 Alkyl amido, amino-PEG 1~4 -C 1-2 Alkylamido C 1-10 Alkylamido, Lysylamido C 1-10 Alkyl amido, or amino-PEG 1~4 -C 1-2 Alkylamido lysylamido C 1-10 It is an alkyl amido;

[0278] B is And, means a part connected to the above A or the above D; and

[0279] LA is hydrogen, or and, n is 14 to 18, and means the site connected to the amino acid residue in A above.

[0280] In the present invention, A is amino C 1-10 Alkyl amido, amino-PEG 1~4 -C 1-3 Alkylamido, Amino-PEG 1~4 -C 1-2 Alkylamido C 1-10 Alkylamido, Lysylamido C 1-10 Alkyl amido, or amino-PEG 1~4 -C 1-2 Alkylamido lysylamido C 1-10 Alkyl amido, amino C 1-2 Alkyl amido, amino C 2-3It may be an alkyl amido, an amino C1 alkyl amido, an amino C2 alkyl amido, or an amino C3 alkyl amido, but is not limited thereto.

[0281] In the present invention, the step (s1) may be performed in the presence of one or more selected from the group consisting of dimethylformamide (DMF), 2-(1H-benzo[d][1,2,3]triazole-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate (V) (HBTU), N,N-diisopropylethylamine (N,N-diisopropylethylamine, DIEA), dichloromethane (DCM), and trifluoroacetic acid (TFA), but is not limited thereto.

[0282] In the present invention, the step (s2) may be performed in the presence of one or more selected from the group consisting of methanol (MeOH), lithium hydroxide (LiOH), and hydrochloric acid (HCl), but is not limited thereto.

[0284] In one embodiment of the present invention, a method for preparing a compound represented by Formula 4 or a pharmaceutically acceptable salt thereof may be as shown in Reaction Scheme 3 below, but is not limited thereto:

[0285] [Reaction Equation 3]

[0286]

[0288] In the present invention, in the above reaction scheme 3, R1, R2, and n may be as described above, and A is amino C 1-10 Alkyl amido, amino-PEG 1~4 -C 1-3 Alkylamido, Amino-PEG 1~4 -C 1-2 Alkylamido C 1-10 Alkylamido, Lysylamido C 1-10 Alkyl amido, or amino-PEG 1~4 -C 1-2 Alkylamido lysylamido C 1-10It may be an alkyl amido, but is not limited thereto. Additionally, Reaction Scheme 3 can be formed by removing hydroxyl and carboxyl protecting groups after a commonly used amide bonding reaction.

[0289] In the present invention, the above reaction formula 3 may be a method for producing a compound represented by formula 6 or a pharmaceutically acceptable salt thereof, but is not limited thereto.

[0290] In one embodiment of the present invention, the compound represented by the chemical formula 11 is,

[0291] (a) a step of synthesizing a compound represented by the following chemical formula 14 by reacting a compound represented by the following chemical formula 13 with a solid phase support into which 2-chlorotrityl chloride has been introduced,

[0292] (b) a step of adding Fmoc-L-glutamic acid 5-tert-butyl ester to the compound formed in step (a) above to form an amide bond,

[0293] (c) a step of adding N-(trifluoroacetyl)pteroic acid to the compound formed in step (b) above to form an amide bond, and

[0294] (d) A method for preparing a compound represented by Formula 2, characterized by comprising one or more steps selected from the group consisting of removing a solid support from the compound formed in step (c) above:

[0295] [Chemical Formula 13]

[0297]

[0298] [Chemical Formula 14]

[0299]

[0300] In the above chemical formulas 13 and 14, respectively, R1 is hydrogen or a carboxyl protecting group, and n is 14 to 18.

[0301] In one embodiment of the present invention, steps (b) and (c) may each be performed independently after deprotecting the Fmoc protector, but are not limited thereto.

[0302] In the present invention, (a) to (d) are each independently Fmoc-L-Lys [Oct-(OtBu)-Glu-(OtBu)-AEEA-AEEA]-OH, N,N-diisopropylethylamine (N,N-diisopylethylamine, DIEA), methyl alcohol (MeOH), dichloromethane (DCM), dimethylformamide (DMF), piperidine, 1-hydroxybenzotriazole (HOBt), O-(benzotriazole-1-yl)- N,N,N',N' -Tetramethyluronium hexafluorophosphate (O -(Benzotriazol-1-yl)- N,N,N',N' It may be performed in the presence of one or more selected from the group consisting of -tetramethyluronium hexafluorophosphate (HBTU), N10-(Trifluoroacetyl)pteroic acid, hydrazine, trifluoroacetic acid (TFA), and diethyl ether, but is not limited thereto.

[0304] In one embodiment of the present invention, the method for preparing the 2-2 compound of Reaction Scheme 3 or a pharmaceutically acceptable salt thereof may be as shown in Reaction Scheme 4 below, but is not limited thereto:

[0305] [Reaction Equation 4]

[0306]

[0307] In the present invention, R1 and n in the above reaction scheme 4 may be as described above, but are not limited thereto. Additionally, reaction scheme 4 may be performed by using a solid phase support to introduce a solid phase, remove amide bonds and protecting groups, and remove the introduced solid phase support.

[0308] In the present invention, the solid support can be used interchangeably with the resin, and in the present invention, the compound of the present invention may replace the solid support into which 2-chlorotritylchloride is introduced with one or more selected from the group consisting of 2-chlorotritylchloride resin, tritylchloride resin, 4-methyltritylchloride resin, and 4-methoxytritylchloride resin, but is not limited thereto.

[0309] In one embodiment of the present invention, the method for preparing a compound or a pharmaceutically acceptable salt thereof is not limited to reaction schemes 1, 2, 3, and 4, and can be modified as necessary at the level of ordinary art.

[0310] In the present invention, the method for preparing compounds 1-2 or a pharmaceutically acceptable salt thereof in Reaction Formula 1, and compounds 1-3 or a pharmaceutically acceptable salt thereof in Reaction Formula 3 may be as shown in Reaction Formula 5 below, but is not limited thereto:

[0311] [Reaction Equation 5]

[0312]

[0313] In the present invention, R1 and R2 in the above reaction formula 5 may be as described above, but are not limited thereto.

[0314] In addition, in the present invention, X may be an amine, and in the case where X is an amine in the reaction, compounds 1-2 or a pharmaceutically acceptable salt thereof in Reaction Scheme 1 and compounds 1-3 or a pharmaceutically acceptable salt thereof in Reaction Scheme 3 may be prepared through a conventional amide bond reaction with carboxylic acids, but are not limited thereto.

[0315] The present invention provides a pharmaceutical composition for the prevention or treatment of cancer, comprising the compound of the present invention or a pharmaceutically acceptable salt thereof as an active ingredient.

[0316] In addition, the present invention provides a pharmaceutical composition for the prevention or treatment of cancer comprising the compound of the present invention, or a pharmaceutically acceptable salt, hydrate, solvate, or isomer thereof as an active ingredient.

[0317] The present invention provides a drug delivery system for the prevention or treatment of cancer, comprising the compound of the present invention or a pharmaceutically acceptable salt thereof as an active ingredient.

[0318] In one embodiment of the present invention, the cancer may be one or more selected from the group consisting of ovarian cancer, breast cancer, kidney cancer, lung cancer, colorectal cancer, and brain cancer, but is not limited thereto.

[0320] The compound according to the present invention can be used to prevent or effectively treat cancer and related diseases by inducing cancer-specific drug delivery and cancer cell death using a folate-drug conjugate utilizing an intelligent drug delivery system based on glucose absorption and metabolism while targeting folate receptor alpha, which is overexpressed in many types of cancer. The cancer diseases include ovarian cancer, breast cancer, kidney cancer, lung cancer, colorectal cancer, brain cancer, or other solid tumors.

[0321] The compound of the present invention may exist in the form of a salt, particularly a pharmaceutically acceptable salt. As the salt, any salt commonly used in the art, such as an acid addition salt formed by a pharmaceutically acceptable free acid, may be used without limitation. The present invention may also include a pharmaceutically acceptable salt of the compound represented by Formula 2 as an active ingredient.

[0322] In the present invention, "pharmaceuticalally acceptable salt" means any organic or inorganic addition salt of said compound at a concentration having a relatively non-toxic and harmless active effect on the patient, such that side effects caused by said salt do not impair the beneficial efficacy of the compound represented by Formula 2. Furthermore, pharmaceutically acceptable salts include salts derived from pharmaceutically acceptable inorganic acids, organic acids, or bases. Additionally, said derivatives include pharmaceutically acceptable salts, pharmaceutically acceptable solvates, pharmaceutically acceptable hydrates, pharmaceutically acceptable anhydrides, pharmaceutically acceptable enantiomers, pharmaceutically acceptable esters, pharmaceutically acceptable polymorphs, pharmaceutically acceptable prodrugs, pharmaceutically acceptable complexes, etc.

[0323] The above free acid may be an organic acid or an inorganic acid. Inorganic acids may include hydrochloric acid, phosphoric acid, sulfuric acid, nitric acid, tartaric acid, etc., and organic acids may include methanesulfonic acid, p-toluenesulfonic acid, acetic acid, trifluoroacetic acid, maleic acid, succinic acid, oxalic acid, benzoic acid, tartaric acid, fumaric acid, manderic acid, propionic acid, citric acid, lactic acid, glycolic acid, gluconic acid, galacturonic acid, glutamic acid, glutaric acid, glucuronic acid, aspartic acid, ascorbic acid, carboxylic acid, vanillic acid, hydroiodic acid, etc., but are not limited to these.

[0324] In addition, pharmaceutically acceptable metal salts can be obtained by conventional methods, such as metal salts formed in free acids using a base. For example, a compound represented by Chemical Formula 2 above can be dissolved in an excess amount of an alkali metal hydroxide or alkaline earth metal hydroxide solution, the undissolved compound salt can be filtered, and the filtrate can be evaporated and dried to obtain a pharmaceutically acceptable metal salt. In this case, it is particularly preferable to prepare a sodium salt, a potassium salt, or a calcium salt as the metal salt.

[0325] Other pharmaceutically unacceptable salts or solvates of the compound represented by Formula 2 may be used as intermediates in the preparation of the compound represented by Formula 2, its pharmaceutically acceptable salts, or solvates.

[0326] In addition, the compound represented by Formula 2 according to the present invention comprises, without limitation, its pharmaceutically acceptable salts as well as solvates such as possible hydrates that can be prepared therefrom and all possible stereoisomers. Solvents and stereoisomers of the compound represented by Formula 2 can be prepared from the compound represented by Formula 2 using methods known in the art.

[0327] In addition, the compound represented by Formula 2 according to the present invention may be prepared in a crystalline or amorphous form, and if prepared in a crystalline form, it may be optionally hydrated or solvated. The present invention may include not only the stoichiometric hydrate of the compound represented by Formula 2 but also compounds containing varying amounts of water. The solvates of the compound represented by Formula 2 according to the present invention include both stoichiometric solvates and non-stoichiometric solvates.

[0328] As used herein, the term “pharmaceuticalally acceptable” means a compound or composition that is suitable for use in contact with tissues of a subject (e.g., human) and within the scope of sound medical judgment, having a reasonable benefit / risk ratio without excessive toxicity, irritation, allergic reaction, or other problems or complications.

[0329] Examples of suitable acids include hydrochloric acid, bromic acid, sulfuric acid, nitric acid, perchloric acid, fumaric acid, maleic acid, phosphoric acid, glycolic acid, lactic acid, salicylic acid, succinic acid, toluene-p-sulfonic acid, tartaric acid, acetic acid, citric acid, methanesulfonic acid, formic acid, benzoic acid, malonic acid, gluconic acid, naphthalene-2-sulfonic acid, benzenesulfonic acid, etc. Acid addition salts can be prepared by conventional methods, for example, by dissolving a compound in an excess amount of an aqueous acid solution and precipitating the salt using a water-miscible organic solvent such as methanol, ethanol, acetone, or acetonitrile. Alternatively, they can be prepared by heating an equal molar amount of the compound and an acid or alcohol in water, followed by drying the mixture by evaporation, or by suction filtration of the precipitated salt.

[0330] Salts derived from suitable bases may include alkali metals such as sodium and potassium, alkaline earth metals such as magnesium, and ammonium, but are not limited thereto. Alkali metal or alkaline earth metal salts can be obtained, for example, by dissolving a compound in an excess amount of alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering the undissolved compound salt, and then evaporating and drying the filtrate. In this case, it is particularly suitable for pharmaceutical purposes to produce sodium, potassium, or calcium salts as metal salts, and the corresponding silver salts can be obtained by reacting an alkali metal or alkaline earth metal salt with a suitable silver salt (e.g., silver nitrate).

[0331] The scope of the compounds of the present invention may include not only pharmaceutically acceptable salts, but also all isomers, hydrates, and solvates that can be prepared by conventional methods.

[0332] In the present invention, “isomer” refers to a compound having the same molecular formula but different physical or chemical properties. In one embodiment of the present invention, the isomer may be one or more selected from the group consisting of structural isomers, enantiomers, optical isomers, stereoisomers, and diastereomers, but is not limited thereto.

[0333] The above compounds may have non-aromatic double bonds and one or more asymmetric centers. Thus, they may occur as racemics and racemic mixtures, a single enantiomer, individual diastereomers, diastereomer mixtures, and cis- or trans-isomers. All such isomer forms are considered.

[0334] The compound represented by Chemical Formula 2 of the present invention may include all compounds exhibiting substantially the same structure, functional group, and function as the compound represented by Chemical Formula 2 among various compounds such as intermediate reactants and secondary products that appear through metabolism or chemical reactions inside or outside the human body.

[0335] The content of the compound represented by Formula 2 in the composition of the present invention can be appropriately adjusted according to the symptoms of the disease, the degree of progression of symptoms, the condition of the patient, etc. For example, it may be 0.0001 to 99.9% by weight or 0.001 to 50% by weight based on the total weight of the composition, but is not limited thereto. The above content ratio is a value based on the dry weight after removing the solvent.

[0336] The pharmaceutical composition according to the present invention may further include a suitable carrier, excipient, and diluent commonly used in the manufacture of pharmaceutical compositions. The excipient may be one or more selected from the group consisting of, for example, diluents, binders, disintegrants, lubricants, adsorbents, humectants, film-coating materials, and controlled-release additives.

[0337] The pharmaceutical composition according to the present invention may be formulated and used in the form of external preparations such as powders, granules, sustained-release granules, enteric granules, liquids, eye drops, oxylic agents, emulsions, suspensions, ethanol tablets, troches, fragrances, limonene adzes, tablets, sustained-release tablets, enteric tablets, sublingual tablets, hard capsules, soft capsules, sustained-release capsules, enteric capsules, pills, tinctures, soft extracts, dry extracts, fluid extracts, injections, capsules, irrigation solutions, warning agents, lotions, pastes, sprays, inhalants, patches, sterile injectable solutions, or aerosols, according to conventional methods, and the external preparations may have formulations such as creams, gels, patches, sprays, ointments, warning agents, lotions, liniments, pastes, or cataplasms.

[0338] Carriers, excipients, and diluents that may be included in the pharmaceutical composition according to the present invention include lactose, dextrose, sucrose, oligosaccharide, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil.

[0339] When formulating, it is prepared using diluents or excipients such as commonly used fillers, extenders, binders, wetting agents, disintegrants, and surfactants.

[0340] Excipients such as corn starch, potato starch, wheat starch, lactose, sucrose, glucose, fructose, D-mannitol, precipitated calcium carbonate, synthetic aluminum silicate, calcium monohydrogen phosphate, calcium sulfate, sodium chloride, sodium bicarbonate, refined lanolin, microcrystalline cellulose, dextrin, sodium alginate, methylcellulose, sodium carboxymethylcellulose, kaolin, urea, colloidal silica gel, hydroxypropyl starch, hydroxypropylmethylcellulose (HPMC) 1928, HPMC 2208, HPMC 2906, HPMC 2910, propylene glycol, casein, calcium lactate, primogel, etc., as additives to tablets, powders, granules, capsules, pills, and lozenges according to the present invention; Gelatin, gum arabic, ethanol, agar powder, cellulose phthalate, carboxymethylcellulose, calcium carboxymethylcellulose, glucose, purified water, sodium casein, glycerin, stearic acid, sodium carboxymethylcellulose, sodium methylcellulose, methylcellulose, microcrystalline cellulose, dextrin, hydroxycellulose, hydroxypropyl starch, hydroxymethylcellulose, refined shellac, starch paste, hydroxypropylcellulose, hydroxypropylmethylcellulose, polyvinyl alcohol, polyvinylpyrrolidone, etc. may be used as binders, and hydroxypropylmethylcellulose, corn starch, agar powder, methylcellulose, bentonite, hydroxypropyl starch, sodium carboxymethylcellulose, sodium alginate, Calcium carboxymethylcellulose, calcium citrate, sodium lauryl sulfate, anhydrous silica, 1-hydroxypropylcellulose, dextran, ion exchange resin, polyvinyl acetate, formaldehyde-treated casein and gelatin, alginic acid, amylose, guar gum, sodium bicarbonate, polyvinylpyrrolidone, calcium phosphate, gelled starch, gum arabic, amylopectin, pectin, sodium polyphosphate, ethylcellulose, sucrose, magnesium aluminum silicate, D-sorbitol solution, hard anhydrous silica, etc. disintegrants;Lubricants such as calcium stearate, magnesium stearate, stearic acid, hydrogenated vegetable oil, talc, lycopodium pods, kaolin, petroleum jelly, sodium stearate, cocoa paste, sodium salicylate, magnesium salicylate, polyethylene glycol (PEG) 4000, PEG 6000, liquid paraffin, hydrogenated soybean oil (Lubri wax), aluminum stearate, zinc stearate, sodium lauryl sulfate, magnesium oxide, macrogol, synthetic aluminum silicate, anhydrous silica, higher fatty acids, higher alcohols, silicone oil, paraffin oil, polyethylene glycol fatty acid ether, starch, sodium chloride, sodium acetate, sodium oleate, dl-leucine, and hard anhydrous silica may be used.

[0341] As additives to the liquid formulation according to the present invention, water, dilute hydrochloric acid, dilute sulfuric acid, sodium citrate, monostearic acid sucroses, polyoxyethylene sorbitol fatty acid esters (tween esters), polyoxyethylene monoalkyl ethers, lanolin ethers, lanolin esters, acetic acid, hydrochloric acid, water ammonia, ammonium carbonate, potassium hydroxide, sodium hydroxide, prolamine, polyvinylpyrrolidone, ethylcellulose, sodium carboxymethylcellulose, etc. may be used.

[0342] In the syrup preparation according to the present invention, a solution of white sugar, other sugars or sweeteners, etc. may be used, and if necessary, flavorings, coloring agents, preservatives, stabilizers, suspending agents, emulsifiers, viscosity enhancers, etc. may be used.

[0343] Purified water may be used in the emulsion according to the present invention, and emulsifiers, preservatives, stabilizers, fragrances, etc. may be used as needed.

[0344] In the suspension agent according to the present invention, suspending agents such as acacia, tragacanthus, methylcellulose, carboxymethylcellulose, sodium carboxymethylcellulose, microcrystalline cellulose, sodium alginate, hydroxypropylmethylcellulose (HPMC), HPMC 1828, HPMC 2906, and HPMC 2910 may be used, and surfactants, preservatives, stabilizers, coloring agents, and fragrances may be used as needed.

[0345] The injectable preparation according to the present invention comprises solvents such as distilled water for injection, 0.9% sodium chloride injection solution, Ringer's injection solution, dextrose injection solution, dextrose + sodium chloride injection solution, PEG, lactated Ringer's injection solution, ethanol, propylene glycol, non-volatile oils—sesame oil, cottonseed oil, peanut oil, soybean oil, corn oil, ethyl oleate, isopropyl myristate, and benzene benzoate; solubilizing agents such as sodium benzoate, sodium salicylate, sodium acetate, urea, urethane, monoethylacetamide, butazolidine, propylene glycol, tween, nijungtinamide, hexamine, and dimethylacetamide; and buffers such as weak acids and their salts (acetic acid and sodium acetate), weak bases and their salts (ammonia and ammonium acetate), organic compounds, proteins, albumin, peptone, and gums. It may include isotonic agents such as sodium chloride; stabilizers such as sodium bisulfite (NaHSO3), carbon dioxide gas, sodium metabisulfite (Na2S2O5), sodium sulfite (Na2SO3), nitrogen gas (N2), and ethylenediaminetetraacetic acid; sulfating agents such as sodium bisulfide 0.1%, sodium formaldehyde sulfoxylate, thiourea, disodium ethylenediaminetetraacetic acid, and sodium bisulfite acetone; non-inflammatory agents such as benzyl alcohol, chlorobutanol, procaine hydrochloride, glucose, and calcium gluconate; and suspending agents such as sodium CMC, sodium alginate, Tween 80, and aluminum monostearate.

[0346] The suppository according to the present invention comprises cocoa dough, lanolin, Witepsol, polyethylene glycol, glycerogelatin, methylcellulose, carboxymethylcellulose, a mixture of stearic acid and oleic acid, Subanal, cottonseed oil, peanut oil, palm oil, cocoa butter + cholesterol, lecithin, lanette wax, glycerol monostearate, Tween or Spandex, Imhausen, monollene (propylene glycol monostearate), glycerin, Adeps solidus, Buytyrum Tego-G, Cebes Pharma 16, hexalide base 95, Cotomar, Hydroccote SP, S-70-XXA, S-70-XX75 (S-70-XX95). Bases such as Hydrokote 25, Hydrokote 711, Idropostal, Massa estrarium (A, AS, B, C, D, E, I, T), Masa-MF, Masupol, Masupol-15, Neosupostal-N, Paramount-B, Suposiro (OSI, OSIX, A, B, C, D, H, L), suppository base type IV (AB, B, A, BC, BBG, E, BGF, C, D, 299), Supostal (N, Es), Wekovi (W, R, S, M, Fs), and Tegestor triglyceride base (TG-95, MA, 57) may be used.

[0347] Solid dosage forms for oral administration include tablets, pills, powders, granules, capsules, etc., and these solid dosage forms are prepared by mixing at least one excipient, such as starch, calcium carbonate, sucrose or lactose, gelatin, etc., with the extract. In addition to simple excipients, lubricants such as magnesium styrate and talc are also used.

[0348] Liquid preparations for oral administration include suspensions, oral liquids, emulsions, and syrups; in addition to commonly used simple diluents such as water and liquid paraffin, they may contain various excipients, such as humectants, sweeteners, flavorings, and preservatives. Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, and suppositories. Non-aqueous solvents and suspensions may include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate.

[0349] The pharmaceutical composition according to the present invention is administered in a pharmaceutically effective amount. In the present invention, "pharmaceutically effective amount" means an amount sufficient to treat a disease with a reasonable benefit / risk ratio applicable to medical treatment, and the effective dose level may be determined according to factors including the type and severity of the patient's disease, drug activity, sensitivity to the drug, time of administration, route of administration and elimination rate, duration of treatment, concurrently used drugs, and other factors well known in the medical field.

[0350] The pharmaceutical composition according to the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, and may be administered sequentially or simultaneously with conventional therapeutic agents, and may be administered as a single or multiple doses. It is important to administer an amount that obtains maximum effect with a minimum amount without side effects by considering all the above-mentioned factors, and this can be easily determined by a person skilled in the art to which the present invention belongs.

[0351] The pharmaceutical composition of the present invention may be administered to an individual by various routes. All modes of administration are expected, for example, oral administration, subcutaneous injection, intraperitoneal administration, intramuscular injection, intrathecal (intradural) injection, sublingual administration, buccal mucosal administration, rectal insertion, vaginal insertion, ocular administration, ear administration, nasal administration, inhalation, spray through the mouth or nose, skin administration, transdermal administration, etc.

[0352] The pharmaceutical composition of the present invention may be formulated into oral or parenteral administration forms in accordance with standard pharmaceutical practices. These formulations may contain, in addition to the active ingredient, pharmaceutically acceptable additives such as carriers, adjuvants, or diluents.

[0353] Suitable carriers include, for example, physiological saline, polyethylene glycol, ethanol, vegetable oil, and isopropyl myristate, and diluents include, for example, lactose, dextrose, sucrose, mannitol, sorbitol, cellulose and / or glycine, but are not limited thereto. In addition, the compounds of the present invention may be dissolved in oils, propylene glycol, or other solvents commonly used in the preparation of injectable solutions. Furthermore, the compounds of the present invention may be formulated into ointments or creams for topical action.

[0354] The preferred dosage of the compound of the present invention varies depending on the patient's condition and body weight, the severity of the disease, the form of the drug, the route of administration, and the duration, but can be appropriately selected by those skilled in the art. However, for a desirable effect, it is preferable to administer the compound of the present invention at a dose of 0.0001 to 100 mg / kg (body weight) per day, preferably 0.001 to 100 mg / kg (body weight). Administration may be performed orally, intravenously, or non-intravenously once a day, once every 2 to 7 days, once a month, or in divided doses.

[0355] Depending on the method of administration, the pharmaceutical composition may contain 0.001 to 99 weight%, preferably 0.01 to 60 weight%, of the compound of the present invention.

[0356] The pharmaceutical composition according to the present invention may be administered to mammals, including rats, mice, livestock, and humans, by various routes. All modes of administration are expected, for example, orally, rectally or intravenously, intramuscularly, subcutaneously, or by intrathecal or cerebrovascular injection.

[0357] The dosage of the pharmaceutical composition of the present invention is determined by the type of active ingredient drug, along with various relevant factors such as the disease to be treated, the route of administration, the patient's age, gender, weight, and the severity of the disease. Specifically, the effective dosage of the composition according to the present invention may vary depending on the patient's age, gender, and weight, and generally, 0.001 to 150 mg, preferably 0.01 to 100 mg per kg of body weight, may be administered daily or every other day, or divided into 1 to 3 doses per day. However, since the dosage may be increased or decreased depending on the route of administration, the severity of the disease, gender, weight, age, etc., the above dosage does not limit the scope of the present invention in any way.

[0358] In the present invention, the term “individual” refers to a subject requiring treatment for a disease and can be used interchangeably with a patient. More specifically, the term “individual” refers to mammals such as humans or non-human primates, mice, rats, dogs, cats, horses, and cattle.

[0359] In the present invention, “administration” means providing a predetermined composition of the present invention to an individual by any appropriate method.

[0360] In the present invention, “prevention” refers to any act of suppressing or delaying the onset of a target disease, “treatment” refers to any act of improving or beneficially altering the target disease and associated metabolic abnormality symptoms through the administration of a pharmaceutical composition according to the present invention, and “improvement” refers to any act of reducing parameters related to the target disease, such as the severity of symptoms, through the administration of a composition according to the present invention.

[0362] In the present invention, the 1-10 (or 1~10) described above refers to 1 to 10, 1 to 9, 1 to 8, 1 to 7, 1 to 6, 1 to 5, 1 to 4, 1 to 3, 1 to 2, 2 to 10, 2 to 9, 2 to 8, 2 to 7, 2 to 6, 2 to 5, 2 to 4, 2 to 3, 3 to 10, 3 to 9, 3 to 8, 3 to 7, 3 to 6, 3 to 5, 3 to 4, 4 to 10, 4 to 9, 4 to 8, 4 to 7, 4 to 6, 4 to 5, 5 to 10, 5 to 9, 5 to 8, 5 to 7, 5 to 6, 6 to 10, 6 It may be up to 9, 6 to 8, 6 to 7, 7 to 10, 7 to 9, 7 to 8, 8 to 10, 8 to 9, or 9 to 10, but is not limited thereto. This is to omit redundant descriptions.

[0363] In the present invention, the 1-4 (or 1~4) described above may be 1 to 4, 1 to 3, 1 to 2, 2 to 4, 2 to 3, or 3 to 4, but is not limited thereto. This is to omit redundant descriptions.

[0364] In the present invention, the 1-3 (or 1~3) described above may be 1 to 3, 1 to 2, or 2 to 3, but is not limited thereto. This is to avoid redundant descriptions.

[0366] Preferred embodiments are presented below to aid in understanding the present invention. However, the following embodiments are provided merely to facilitate a better understanding of the invention, and the scope of the invention is not limited by the following embodiments.

[0368] [Example]

[0370] Example 1. Preparation of (2R,3S,4S,5R)-6-(2-(2-(4-(4-(((2-amino-4-oxo-3,4-dihydropteridine-6-yl)methyl)amino)benzamido)-4-carboxybutanemido)acetamido)-4-(((((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[di]pyrano[3',4':6,7]yldolizino[1,2-b]quinoline-1-yl)carbamoyl)oxy)methyl)phenoxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylic acid

[0372] In this Example 1, through the process of Preparation Examples 1-1 to 1-8 below, the formula represented by [Chemical Formula 3] below (2R,3S,4S,5R)-6-(2-(2-(4-(4-(((2-amino-4-oxo-3,4-dihydropteridine-6-yl)methyl)amino)benzamido)-4-carboxybutanamido)acetamido)-4-(((((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[di]pyrano[3',4':6,7]yldolizino[1,2-b]quinoline-1-yl)carbamoyl)oxy)methyl)phenoxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylic acid was prepared:

[0373] [Chemical Formula 3]

[0374] .

[0376] <Preparation Example 1-1> Preparation of (2S,3R,4S,5S,6R)-2-(4-(hydroxymethyl)-2-nitrophenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate

[0378] In Preparation Example 1-1, (2S,3R,4S,5S,6R)-2-(4-(hydroxymethyl)-2-nitrophenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate represented by the following [Chemical Formula 15] was prepared:

[0379] [Chemical Formula 15]

[0380] .

[0382] 4-(hydroxymethyl)-2-nitrophenol (1 equiv.), (2R,3R,4S,5S,6R)-2-bromo-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (1 equiv.), silver oxide (1.1 equiv.) were dissolved in anhydrous acetonitrile and stirred at room temperature for 4 hours. After the reaction was completed, the solution obtained by filtration was concentrated under reduced pressure, and the obtained product was washed and filtered using ether to obtain the yellow title compound in 60% yield.

[0384] 1 H NMR (400 MHz, CDCl3) δ 7.84 (d, J = 2.1 Hz, 1H), 7.56 (dd, J = 8.6, 2.2 Hz, 1H), 7.40 (d, J = 8.6 Hz, 1H), 5.42 - 5.29 (m, 4H), 5.22 (d, J = 6.9 Hz, 1H), 4.75 (d, J = 5.5 Hz, 2H), 4.23 (d, J = 8.9 Hz, 1H), 3.77 (s, 3H), 2.15 (s, 3H), 2.11 - 2.05 (m, 6H).

[0386] <Preparation Example 1-2> Preparation of (2R,3S,4S,5R)-2-(methoxycarbonyl)-6-(2-nitro-4-((((4-nitrophenoxy)carbamoyl)oxy)methyl)phenoxy)tetrahydro-2H-pyran-3,4,5-triyl triacetate

[0387] In Preparation Examples 1-2, (2R,3S,4S,5R)-2-(methoxycarbonyl)-6-(2-nitro-4-((((4-nitrophenoxy)carbamoyl)oxy)methyl)phenoxy)tetrahydro-2H-pyran-3,4,5-triyl triacetate represented by the following [Chemical Formula 16] was prepared:

[0388] [Chemical Formula 16]

[0389] .

[0391] The compound prepared in Preparation Example 1-1 was dissolved in dichloromethane (DCM), then 4-nitrophenyl carbonochloridate (1.1 equiv.) and pyridine (0.1 equiv.) were added and stirred at room temperature for 20 minutes. After the reaction was finished, the solvent was concentrated under reduced pressure to obtain the yellow title compound in 87% yield.

[0393] <Preparation Example 1-3> Preparation of (3R,4S,5S,6R)-2-(4-(((((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[di]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-1-yl)carbamoyl)oxy)methyl)-2-nitrophenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate

[0395] In Preparation Examples 1-3, (3R,4S,5S,6R)-2-(4-(((((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[di]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-1-yl)carbamoyl)oxy)methyl)-2-nitrophenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate represented by the following [Chemical Formula 17] was prepared:

[0396] [Chemical Formula 17]

[0397] .

[0399] The compound (1 equiv) prepared in Preparation Example 1-2 was dissolved in dimethylformamide (DMF), then exatecan (1 equiv) and DIEA (3 equiv) were added and stirred at room temperature for 2 hours. Water was added to the reaction mixture and the reaction was terminated. The reaction mixture was extracted three times with dichlorometal (DCM), then the organic layer was dried over sodium sulfate and concentrated under reduced pressure. The obtained material was separated from the column, and the eluent was concentrated under reduced pressure to obtain the brown title compound in a yield of 44%.

[0400] MS calcd for C 45 H 43 FN4O18 = 946.24, found = 947.15 [M+H + ]

[0402] <Preparation Example 1-4> Preparation of (3R,4S,5S,6R)-2-(2-amino-4-(((((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[di]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-1-yl)carbamoyl)oxy)methyl)phenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate

[0404] In Preparation Examples 1-4, (3R,4S,5S,6R)-2-(2-amino-4-(((((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[di]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-1-yl)carbamoyl)oxy)methyl)phenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate represented by the following [Chemical Formula 18] was prepared:

[0405] [Chemical Formula 18]

[0406] .

[0408] The compound (1 equiv.) prepared in Preparation Examples 1-3 was dissolved in methanol (MeOH) / ethanol (EtOH) / water (H2O) (3:3:1 v / v), then iron (Fe) (3 equiv) and ammonium chloride (10 equiv) were added, and the mixture was stirred at room temperature for 8 hours. The reaction mixture was filtered and concentrated under reduced pressure. The concentrated compound was washed with dichloromethane (DCM), filtered, and concentrated under reduced pressure to obtain the brown title compound in a yield of 52%.

[0409] MS calcd for C 45 H 45 FN4O 16 = 916.28, found = 917.20 [M+H + ]

[0411] <Preparation Example 1-5> Preparation of (3R,4S,5S,6R)-2-(2-(2-((tert-butoxycarbonyl)amino)acetamido)-4-(((((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexydro-1H,12H-benzo[di]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-1-yl)carbamoyl)oxy)methyl)phenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triacetate

[0413] In Preparation Examples 1-5, (3R,4S,5S,6R)-2-(2-(2-((tert-butoxycarbonyl)amino)acetamido)-4-(((((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexydro-1H,12H-benzo[di]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-1-yl)carbamoyl)oxy)methyl)phenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triacetate represented by the following [Chemical Formula 19] was prepared:

[0414] [Chemical Formula 19]

[0415] .

[0417] The compound (1 equiv) prepared in Preparation Examples 1-4 was dissolved in dimethylformamide (DMF), then (t-butoxycarbonyl)glycine (1 equiv.), O-(benzotriazole-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HBTU) (1 equiv.), and N,N-diisopropylethylamine (DIEA) (3 equiv.) were added and stirred at room temperature for 8 hours. Water was added to the reaction mixture and the reaction was terminated. The reaction mixture was extracted three times with dichloromethane (DCM), after which the organic layer was dried over sodium sulfate and concentrated under reduced pressure. The obtained material was separated by column chromatography, and the eluent was evaporated to obtain the brown title compound in a yield of 38%.

[0418] MS calcd for C 52 H 56 FN5O 19 = 1073.36, found = 1074.25 [M+H + ]

[0420] <Preparation Example 1-6> Preparation of (3R,4S,5S,6R)-2-(2-(2-aminoacetamido)-4-(((((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[di]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-1-yl)carbamoyl)oxy)methyl)phenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triacetate

[0422] In Preparation Examples 1-6, (3R,4S,5S,6R)-2-(2-(2-aminoacetamido)-4-(((((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[di]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-1-yl)carbamoyl)oxy)methyl)phenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triacetate represented by the following [Chemical Formula 20] was prepared:

[0423] [Chemical Formula 20]

[0424] .

[0426] The compound (1 equiv.) prepared in Preparation Examples 1-5 was dissolved in dichloromethane (DCM), then trifluoroacetic acid (TFA) (3 equiv.) was added and stirred at room temperature for 3 hours. After the reaction was finished, the reaction product was concentrated under reduced pressure to obtain the brown title compound in a yield of 88%.

[0427] MS calcd for C 47 H 48 FN5O 17 = 973.30, found = 974.20 [M+H + ]

[0429] <Preparation Example 1-7> Preparation of N2-(4-(((2-amino-4-oxo-3,4-dihydrotheridine-6-yl)methyl)amino)benzoyl)-N5-(2-((5-(((((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[di]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-1-yl)carbamoyl)oxy)methyl-2-(((3R,4S,5S,6R)-3,4,5-triacetoxy-6-(methoxycarbonyl)tetrahydro-2H-pyran-2-yl)oxy)phenyl)amino)-2-oxoethyl)glutamine

[0431] In Preparation Examples 1-7, the formula is represented by the following [Chemical Formula 21]. N2-(4-(((2-amino-4-oxo-3,4-dihydrotheridine-6-yl)methyl)amino)benzoyl)-N5-(2-((5-((((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[di]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-1-yl)carbamoyl)oxy)methyl-2-(((3R,4S,5S,6R)-3,4,5-triacetoxy-6-(methoxycarbonyl)tetrahydro-2H-pyran-2-yl)oxy)phenyl)amino)-2-oxoethyl)glutamine was prepared:

[0432] [Chemical Formula 21]

[0433] .

[0435] 1 mL of dimethyl sulfoxide (DMSO) was added to folic acid (1 equiv.), N,N'-dicyclohexylcarbodiimide (2 equiv.), N-hydroxysuccinimide (1 equiv.), and pyridine (2 equiv.), and stirred at room temperature for 4 hours. Then, the compound prepared in Preparation Example 1-6 (1 equiv.) was added and stirred at room temperature for 12 hours. Water was added to the reaction mixture to terminate the reaction. The reaction mixture was extracted three times with dichloromethyl (DCM), and the organic layer was dried over sodium sulfate and concentrated under reduced pressure. The obtained material was separated from the column, and the eluent was concentrated under reduced pressure to obtain the brown title compound in a yield of 16%.

[0436] MS calcd for C 66 H 65 FN 12 O 22 = 1396.43, found 1398.10 [M+H + ]

[0438] <Preparation Example 1-8> Preparation of (2R,3S,4S,5R)-6-(2-(2-(4-(4-(((2-amino-4-oxo-3,4-dihydropteridine-6-yl)methyl)amino)benzamido)-4-carboxybutanemid)acetamido)-4-(((((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[di]pyrano[3',4':6,7]yldolizino[1,2-b]quinoline-1-yl)carbamoyl)oxy)methyl)phenoxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylic acid

[0440] The compound (1 equiv.) prepared in Preparation Examples 1-7 was dissolved in methanol (MeOH), 1N lithium hydroxide (LiOH) (4 equiv.) was added, and the mixture was stirred at room temperature for 2 hours. 1N hydrochloric acid (HCl) was added to the reaction mixture to adjust the pH to 6, and then concentrated under reduced pressure. The obtained substance was separated from the column, and the eluent was evaporated to obtain a brown (2R,3S,4S,5R)-6-(2-(2-(4-(4-(((2-amino-4-oxo-3,4-dihydropteridine-6-yl)methyl)amino)benzamido)-4-carboxybutanemido)acetamido)-4-(((((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[di]pyrano[3',4':6,7]yldolizino[1,2-b]quinoline-1-yl)carbamoyl)oxy)methyl)phenoxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylic acid compound in 33% yield obtained.

[0441] MS calcd for C 59 H 57 FN 12 O 19 = 1256.38, found = 1257.50 [M+H + ]

[0443] Example 2. 1-(4-(((2-amino-4-oxo-3,4-dihydropteridine-6-yl)methyl)amino)phenyl)-8-((2-((2-((6-carboxy-3,4,5-trihydroxytetrahydro-2H-pyran-2-yl)oxy)-5-(((((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexa Hydro-1H,12H-benzo[di]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-1-yl)carbamoyl)oxy)methyl)phenyl)amino)-2-oxoethyl)carbamoyl)-1,6,14,23,32,37-hexaoxo-16,19,25,28-tetraoxo-2,7,13,22,31,36-hexaazatripentacontan-3,35,53-preparation of tricarboxylic acid

[0445] In Example 2, through the processes of Preparation Examples 2-1 to 2-4 below, 1-(4-(((2-amino-4-oxo-3,4-dihydropteridine-6-yl)methyl)amino)phenyl)-8-((2-((2-((6-carboxy-3,4,5-trihydroxytetrahydro-2H-pyran-2-yl)oxy)-5-(((((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexa Hydro-1H,12H-benzo[di]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-1-yl)carbamoyl)oxy)methyl)phenyl)amino)-2-oxoethyl)carbamoyl)-1,6,14,23,32,37-hexaoxo-16,19,25,28-tetraoxo-2,7,13,22,31,36-hexaazatripentacontan-3,35,53-tricarboxylic acid was prepared:

[0446] [Chemical Formula 4]

[0447] .

[0449] <Preparation Example 2-1> Preparation of Intermediate Compound 2-2 using Solid-Phase Synthesis

[0451] In Preparation Example 2-1, intermediate compound 2-2 was prepared through the following [Reaction Scheme 6].

[0452] [Reaction Equation 6]

[0453]

[0455] The intermediate compound of the present invention was synthesized using the solid-phase amino acid synthesis method (Umbarger. HE, Ann. Rev. Biochem. 1978, 47, 533-606). The C-terminal initiation of the compound was 2-chlorotritylchloride resin (100-200 mesh), 1% DVB Novabiochem ®To prepare a compound consisting of a COOH carboxyl group terminus, Fmoc-L-Lys [Oct-(OtBu)-Glu-(OtBu)-AEEA-AEEA-OH] and N,N-diisopropylethylamine (DIEA) were added to a glass vial using (100 mg, 1.33 mmol / g) and stirred for 30 to 120 minutes, then the reaction was terminated after washing 3 times with a methyl alcohol (MEOH) / dichloromethane (DCM) / N,N-diisopropylethylamine (DIEA) (17 / 2 / 1, v / v) solution. After the reaction was completed, the resin was transferred to a Torviq reaction vessel polypropylene syringe and washed twice each using dimethylformamide (DMF), methyl alcohol (MeOH), dichloromethane (DCM), and finally dimethylformamide (DMF).

[0456] For the second step amide bonding, the reaction was carried out twice for 10 minutes each at room temperature using dimethylformamide (DMF) (1 mL / 100 mg resin) with 20% (v / v) piperidine added for the Fmoc protecting group removal reaction, followed by washing the resin twice each with dimethylformamide (DMF), methyl alcohol (MeOH), dichloromethane (DCM), and finally dimethylformamide (DMF), after which hydroxybenzotriazole (HOBt) and O-(benzotriazole-1-yl)- N,N,N',N' -Tetramethyluronium hexafluorophosphate (O -(Benzotriazol-1-yl)- N,N,N',N'-tetramethyluronium hexafluorophosphate (HBTU), N,N-diisopropylethylamine (DIEA), and Fmoc-Glu-(OtBu) were stirred in a dimethylformamide (DMF) solution for 2 hours. The reaction-completed resin was washed twice each using dimethylformamide (DMF), methyl alcohol (MeOH), dichloromethane (DCM), and finally dimethylformamide (DMF). The amide synthesis step in the next stage was also synthesized using N10-(Trifluoroacetyl)pteroic acid in the same manner as the peptide synthesis step in the second stage, and was completed through the washing process described above.

[0457] After the reaction was completed, 1 mL of dimethylformamide (DMF) containing 2% hydrazine was added to the N-trifluoroacetyl protecting group to complete the deprotection reaction, and the compound bound to the crude resin obtained through the washing process above was reacted for 30 minutes using a dichloromethane (DCM) solution containing 2% trifluoroacetic acid (TFA) to separate the compound from the resin, and then the compound was precipitated with cooled diethyl ether to obtain the intermediate compound 2-2 of the title.

[0458] MS calcd for C 72 H 116 N 12 O 19 = 1452.85, found = 1454.55 (M+H + )

[0460] <Preparation Example 2-2> Tri-tert-butyl 1-(4-(((2-amino-4-oxo-3,4-dihydropteridine-6-yl)methyl)amino)phenyl)-8-((2-((5-(((((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro -1H,12H-benzo[di]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-1-yl)carbamoyl)oxy)methyl)-2-((3,4,5-triacetoxy-6-(methoxycarbonyl)tetrahydro-2H-pyran-2-yl)oxy)phenyl)amino)-2-oxoethyl)carbamoyl)-1,6,14,23,32,37-hexaoxo-16,19,25,28-tetraoxa-2,7,13,22,31,36-hexaazatripentacontan-3,35,53-tricarboxylate preparation

[0462] In Preparation Example 2-2, Tri-tert-butyl 1-(4-(((2-amino-4-oxo-3,4-dihydropteridine-6-yl)methyl)amino)phenyl)-8-((2-((5-(((((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro -1H,12H-benzo[di]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-1-yl)carbamoyl)oxy)methyl)-2-((3,4,5-triacetoxy-6-(methoxycarbonyl)tetrahydro-2H-pyran-2-yl)oxy)phenyl)amino)-2-oxoethyl)carbamoyl)-1,6,14,23,32,37-hexaoxo-16,19,25,28-tetraoxa-2,7,13,22,31,36-hexaazatripentacontan-3,35,53-tricarboxylate was prepared:

[0463] [Chemical Formula 22]

[0464] .

[0466] The compound prepared in Preparation Example 2-1 (1 equiv.) was dissolved in dimethylformamide (DMF), and then 2-(1H-benzo[d][1,2,3]triazole-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate (V) (HBTU) (1 equiv.) and N,N-diisopropylethylamine (DIEA) (3 equiv.) were added. After stirring for 30 minutes, compound 6 (1 equiv.) was added and stirred for 4 hours. Water was added to the reaction mixture to terminate the reaction. The reaction mixture was extracted three times with dichloromethane (DCM), and the organic layer was dried with sodium sulfate and concentrated under reduced pressure. The obtained material was separated from the column, and the eluent was evaporated to obtain the title compound in a yield of 54%.

[0467] MS calcd for C 119 H 162 FN 17 O 35 = 2408.14, found = 1205.50 [M+2H2+ ]

[0469] <Preparation Example 2-3> 1-(4-(((2-amino-4-oxo-3,4-dihydropteridine-6-yl)methyl)amino)phenyl)-8-((2-((5-(((((1S,9S)-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[di]pyrano[3',4':6,7]indoleizino[1,2-b]quinoline-1-yl)carbamoyl)oxy)methyl)-2-((3,4,5- Triacetoxy-6-(methoxycarbonyl)tetrahydro-2H-pyran-2-yl)oxy)phenyl)amino)-2-oxoethyl)carbamoyl)-1,6,14,23,32,37-hexaoxo-16,19,25,28-tetraoxa-2,7,13,22,31,36-hexaazatripentacontan-3,35,53-preparation of tricarboxylic acid

[0471] In Preparation Examples 2-3, the 1-(4-(((2-amino-4-oxo-3,4-dihydropteridine-6-yl)methyl)amino)phenyl)-8-((2-((5-(((((1S,9S)-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[di]pyrano[3',4':6,7]indoleizino[1,2-b]quinoline-1-yl)carbamoyl)oxy)methyl)-2-((3,4,5- Triacetoxy-6-(methoxycarbonyl)tetrahydro-2H-pyran-2-yl)oxy)phenyl)amino)-2-oxoethyl)carbamoyl)-1,6,14,23,32,37-hexaoxo-16,19,25,28-tetraoxa-2,7,13,22,31,36-hexaazatripentacontan-3,35,53-tricarboxylic acid was prepared:

[0472] [Chemical Formula 23]

[0473] .

[0475] The compound prepared in Preparation Example 2-2 was dissolved in 1 ml of dichloromethane (DCM), then trifluoroacetic acid (TFA) (4 equiv.) was added and stirred at room temperature for 3 hours. After the reaction was finished, the compound was concentrated under reduced pressure to obtain the title compound.

[0476] MS calcd for C 107 H 138 FN 17 O 35 = 2239.95, found = 1121.30 [M+2H 2+ ]

[0478] <Preparation Example 2-4> 1-(4-(((2-amino-4-oxo-3,4-dihydropteridine-6-yl)methyl)amino)phenyl)-8-((2-((2-((6-carboxy-3,4,5-trihydroxytetrahydro-2H-pyran-2-yl)oxy)-5-(((((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexa Hydro-1H,12H-benzo[di]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-1-yl)carbamoyl)oxy)methyl)phenyl)amino)-2-oxoethyl)carbamoyl)-1,6,14,23,32,37-hexaoxo-16,19,25,28-tetraoxo-2,7,13,22,31,36-hexaazatripentacontan-3,35,53-preparation of tricarboxylic acid

[0480] The compound (1 equiv.) prepared in Preparation Examples 2-3 was dissolved in methanol (MeOH), 1N lithium hydroxide (LiOH) (4 equiv.) was added, and the mixture was stirred at room temperature for 2 hours. 1N hydrochloric acid (HCl) was added to the reaction mixture to adjust the pH to 6, and the mixture obtained by concentrating under reduced pressure was separated from the column. The eluent was then evaporated to obtain 1-(4-(((2-amino-4-oxo-3,4-dihydropteridine-6-yl)methyl)amino)phenyl)-8-((2-((2-((6-carboxy-3,4,5-trihydroxytetrahydro-2H-pyran-2-yl)oxy)-5-(((((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexa Hydro-1H,12H-benzo[di]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-1-yl)carbamoyl)oxy)methyl)phenyl)amino)-2-oxoethyl)carbamoyl)-1,6,14,23,32,37-hexaoxo-16,19,25,28-tetraoxo-2,7,13,22,31,36-hexaazatripentacontan-3,35,53-tricarboxylic acid compound was obtained in a yield of 43%.

[0481] MS calcd for C 100 H 130 FN 17 O 32 = 2099.91, found =1051 [M+2H 2+ ]

[0483] Example 3. 19-(4-(((2-amino-4-oxo-3,4-dihydropteridine-6-yl)methyl)amino)benzamido)-1-((2-(((3R,4S,5S,6R)-6-carboxy-3,4,5-trihydroxytetrahydro-2H-pyran-2-yl)oxy)-5-(((((1S,9S)-9-ethyl-5-fluoro-9-hydroxy -4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[di]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-1-yl)carbamoyl)oxy)methyl)phenyl)amino)-1,4,16-trioxo-6,9,12-trioxa-3,15-diazycosan-20-preparation of ic acid

[0485] In Example 3, 19-(4-(((2-amino-4-oxo-3,4-dihydropteridine-6-yl)methyl)amino)benzamido)-1-((2-(((3R,4S,5S,6R)-6-carboxy-3,4,5-trihydroxytetrahydro-2H-pyran-2-yl)oxy)-5-(((((1S,9S)-9-ethyl-5-fluoro-9-hydroxy -4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[di]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-1-yl)carbamoyl)oxy)methyl)phenyl)amino)-1,4,16-trioxo-6,9,12-trioxa-3,15-diazaicosan-20-ic acid was prepared:

[0486] [Chemical Formula 5]

[0487] .

[0489] The title compound was obtained using the same method as in Preparation Examples 1-7 of Example 1, except that folic acid-NH-PEG3-CH2CO2H was used instead of folic acid.

[0490] MS calcd for C 74 H 80 FN 13 O 26 (M+H) + : 1586, found 1587

[0492] Example 4. 1-(4-(((2-amino-4-oxo-3,4-dihydropteridine-6-yl)methyl)amino)phenyl)-17-((2-((2-(((3R,4S,5S,6R)-6-carboxy-3,4,5-trihydroxytetrahydro-2H-pyran-2-yl)oxy)-5-(((((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15- Hexahydro-1H,12H-benzo[di]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-1-yl)carbamoyl)oxy)methyl)phenyl)amino)-2-oxoethyl)carbamoyl)-1,6,15,23,32,41,46-heptaoxo-10,13,25,28,34,37-hexaoxo-2,7,16,22,31,40,45-heptaazadohexacontan-3,44,62-preparation of tricarboxylic acid

[0494] In Example 4, 1-(4-(((2-amino-4-oxo-3,4-dihydropteridine-6-yl)methyl)amino)phenyl)-17-((2-((2-(((3R,4S,5S,6R)-6-carboxy-3,4,5-trihydroxytetrahydro-2H-pyran-2-yl)oxy)-5-(((((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15- Hexahydro-1H,12H-benzo[di]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-1-yl)carbamoyl)oxy)methyl)phenyl)amino)-2-oxoethyl)carbamoyl)-1,6,15,23,32,41,46-heptaoxo-10,13,25,28,34,37-hexaoxo-2,7,16,22,31,40,45-heptaazadohexacontan-3,44,62-tricarboxylic acid was prepared:

[0495] [Chemical Formula 6]

[0496]

[0497] The title compound was obtained using the same method as in Preparation Example 2-1 of Example 2, except that protected folic acid-NH-PEG2-CH2CO2H was used instead of protected folic acid.

[0498] MS calcd for C 106 H 141 FN 18 O 35 (M+H) + : 2246(1123), found 1124

[0500] Example 5. 20-(4-(((2-amino-4-oxo-3,4-dihydropteridine-6-yl)methyl)amino)benzamido)-1-((2-(((2R,3R,4S,5S,6R)-6-carboxy-3,4,5-trihydroxytetrahydro-2H-pyran-2-yl)oxy)-5-(((((1S,9S)-9-ethyl-5-fluoro-9-hydroxy -4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[di]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-1-yl)carbamoyl)oxy)methyl)phenyl)amino)-1,5,17-trioxo-7,10,13-trioxa-4,16-diazahenicosan-21-preparation of icic acid

[0502] In Example 5, 20-(4-(((2-amino-4-oxo-3,4-dihydropteridine-6-yl)methyl)amino)benzamido)-1-((2-(((2R,3R,4S,5S,6R)-6-carboxy-3,4,5-trihydroxytetrahydro-2H-pyran-2-yl)oxy)-5-(((((1S,9S)-9-ethyl-5-fluoro-9-hydroxy -4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[di]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-1-yl)carbamoyl)oxy)methyl)phenyl)amino)-1,5,17-trioxo-7,10,13-trioxa-4,16-diazahenicosan-21-ic acid was prepared:

[0503] [Chemical Formula 7]

[0504] .

[0506] The title compound was obtained using the same preparation method as in Example 3, except that 3-((((9H-fluorene-9-yl)methoxy)carbonyl)amino)propanoic acid was used instead of (t-butoxycarbonyl)glycine.

[0507] MS calcd for C 68 H 74 FN 13 O 23 (M+H) + : 1460, found 1461

[0509] Example 6. (2R,3S,4S,5R)-6-(2-(3-(4-(4-(((2-amino-4-oxo-3,4-dihydropteridine-6-yl)methyl)amino)benzamido)-4-carboxybutanamido)butanamido)propaneamido)-4-(((((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[di]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-1-yl)carbamoyl)oxy)methyl)phenoxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylic acid preparation

[0511] In this Example 6, the formula is represented by the following [Chemical Formula 8]. (2R,3S,4S,5R)-6-(2-(3-(4-(4-(((2-amino-4-oxo-3,4-dihydropteridine-6-yl)methyl)amino)benzamido)-4-carboxybutanamido)butanamido)propaneamido)-4-(((((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[di]pyrano[3',4':6,7]indoleizino[1,2-b]quinoline-1-yl)carbamoyl)oxy)methyl)phenoxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylic acid Manufactured:

[0512] [Chemical Formula 8]

[0513] .

[0514] The title compound was obtained using the same method, except that protected folic acid-NH-(CH2)3CO2H was used instead of protected folic acid-NH-PEG3-CH2CO2H of Example 5.

[0515] MS calcd for C 64 H 66 FN 13 O 20 (M+H) + : 1356, found 1357

[0517] Example 7. Results of Anticancer Activity (Cell Proliferation Assay) Evaluation

[0519] The anticancer activity (GI) was evaluated by measuring the degree of cell proliferation inhibition of the compound prepared in the above example using the KB cell line (ATCC CRL-3596) (Folate receptor positive; FR positive) and a cell proliferation assay system. 50 Evaluates ).

[0521] The specific evaluation method is as follows.

[0522] Approximately 4,000 KB cells in RPMI1640 (Gibco, 27012-021) medium containing 10% Fetal Bovine Serum, 5% CO 2, Incubated at 37°C for 24 hours. The test substance of the example to be evaluated and exatecan were treated at concentrations of 10 points. After 72 hours, the CCK-8 kit solution Cell proliferation inhibition was determined by treatment and measuring the response value at 450 nm absorbance, GI 50 It was derived as a value. Anticancer activity (GI) measured by the degree of cell proliferation inhibition of each test substance 50 ) exhibited excellent activity of 10 μM or less.

[0523] item Compound represented by Chemical Formula 4 Compound represented by Chemical Formula 7 GIVE 50 , uM 1.04 0.73

[0524] The foregoing description of the present invention is for illustrative purposes only, and those skilled in the art will understand that other specific forms can be easily modified without altering the technical spirit or essential features of the present invention. Therefore, the embodiments described above should be understood as illustrative in all respects and not restrictive.

Claims

Claim 1 A compound represented by the following chemical formula 2, or a pharmaceutically acceptable salt thereof: [Chemical Formula 2] In the above chemical formula 2, A-LA is or and * represents the region connected to C in Chemical Formula 2, means the part connected to B, and B is and * indicates the part connected to A-LA, refers to the part connected to D, and D is And, means the part connected to B. Claim 2 delete Claim 3 delete Claim 4 The compound of claim 1, or a pharmaceutically acceptable salt thereof, characterized in that the compound represented by Formula 2 is selected from the group consisting of the following: (2) 1-(4-(((2-amino-4-oxo-3,4-dihydropteridine-6-yl)methyl)amino)phenyl)-8-((2-((2-((6-carboxy-3,4,5-trihydroxytetrahydro-2H-pyran-2-yl)oxy)-5-(((((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexa Hydro-1H,12H-benzo[di]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-1-yl)carbamoyl)oxy)methyl)phenyl)amino)-2-oxoethyl)carbamoyl)-1,6,14,23,32,37-hexaoxo-16,19,25,28-tetraoxo-2,7,13,22,31,36-hexaazatripentacontan-3,35,53-tricarboxylic acid; and (5) 20-(4-(((2-amino-4-oxo-3,4-dihydropteridine-6-yl)methyl)amino)benzamido)-1-((2-(((2R,3R,4S,5S,6R)-6-carboxy-3,4,5-trihydroxytetrahydro-2H-pyran-2-yl)oxy)-5-(((((1S,9S)-9-ethyl-5-fluoro-9-hydroxy -4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[di]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-1-yl)carbamoyl)oxy)methyl)phenyl)amino)-1,5,17-trioxo-7,10,13-trioxa-4,16-diazahenicosan-21-ican. Claim 5 A compound or a pharmaceutically acceptable salt thereof, characterized in that, in claim 1, the compound represented by Chemical Formula 2 is selected from the group consisting of Chemical Formulas 4 and 7. [Chemical Formula 4] [Chemical Formula 7] Claim 6 (s1) a step of adding a compound represented by the following chemical formula 10 to a compound represented by the following chemical formula 9 to form an amide bond; and (s2) a step of deprotecting a hydroxyl protecting group or a carboxyl protecting group of the compound formed in step (s1), comprising a method for preparing a compound represented by the following chemical formula 2: [Chemical Formula 9] [Chemical Formula 10] [Chemical Formula 2] In the above chemical formula 9, R4 is And, represents the region connected to C of Chemical Formula 9; in the above Chemical Formula 10, A-LA is And, represents the site connected to C in Chemical Formula 10, R1 is a carboxyl protecting group, said carboxyl protecting group is -CH3, R2 is a hydroxyl protecting group, said hydroxyl protecting group is an acetyl group (Ac); in the above Chemical Formula 2, A-LA is and * represents the region connected to C in Chemical Formula 2, means the part connected to B, and B is and * refers to the part connected to the above A-LA, means the part connected to the above D; D is And, means the part connected to B. Claim 7 delete Claim 8 (s1) a step of adding a compound represented by the following chemical formula 12 to a compound represented by the following chemical formula 11 to form an amide bond, and (s2) a step of deprotecting a hydroxyl protecting group or a carboxyl protecting group of the compound formed in step (s1), comprising a method for preparing a compound represented by the following chemical formula 2: [Chemical Formula 11] [Chemical Formula 12] [Chemical Formula 2] In the above chemical formula 11, R1 is a carboxyl protecting group, the carboxyl protecting group is -C(CH3)3, and n is 16; in the above chemical formula 12, A is is, represents the site connected to C in Chemical Formula 12, R1 is a carboxyl protecting group, said carboxyl protecting group is -CH3, R2 is a hydroxyl protecting group, said hydroxyl protecting group is an acetyl group (Ac); in the above Chemical Formula 2, A-LA is and * represents the region connected to C in Chemical Formula 2, means the part connected to B, and B is and * indicates the part connected to A-LA, refers to the part connected to D, and D is And, means the part connected to B. Claim 9 delete Claim 10 A method for preparing a compound represented by Chemical Formula 2 according to claim 8, wherein the compound represented by Chemical Formula 11 is prepared by a method comprising one or more steps selected from the group consisting of: (a) reacting a compound represented by Chemical Formula 13 below with a solid-phase support to which 2-chlorotrityl chloride has been introduced to form a compound represented by Chemical Formula 14 below; (b) adding a fluorenylmethoxycarbonyl protecting group (9-Fluorenylmethyloxycarbonyl, Fmoc)-L-glutamic acid 5-tert-butyl ester to the compound formed in step (a) to form an amide bond; (c) adding N-(trifluoroacetyl)pteroic acid to the compound formed in step (b) to form an amide bond; and (d) removing the solid-phase support from the compound formed in step (c). [Chemical Formula 13] [Chemical Formula 14] In the above chemical formulas 13 and 14, independently R1 is a carboxyl protecting group, the carboxyl protecting group is -C(CH3)3, and n is 16. Claim 11 A method for preparing a compound represented by Formula 2, wherein, in claim 10, steps (b) and (c) are each performed independently after deprotecting a fluorenylmethoxycarbonyl protecting group (9-Fluorenylmethyloxycarbonyl, Fmoc). Claim 12 A pharmaceutical composition for the prevention or treatment of cancer, comprising as an active ingredient a compound of any one of claims 1, 4, and 5, or a pharmaceutically acceptable salt thereof. Claim 13 A pharmaceutical composition for the prevention or treatment of cancer according to claim 12, characterized in that the cancer is one or more selected from the group consisting of ovarian cancer, breast cancer, kidney cancer, lung cancer, colorectal cancer, and brain cancer. Claim 14 delete Claim 15 delete

Images