Pharmaceutical composition for preventing or treating cancer

Abstract

The present invention relates to a novel oligopeptide AQTGTGKT analog compound, and more specifically, to an analog compound of oligopeptide AQTGTGKT that exhibits excellent anti-cancer effects and is stable in human blood, a pharmaceutical composition containing the same as an active ingredient, and a method for preparing the same. The analog compound and pharmaceutical composition containing the same as an active ingredient according to the present invention not only have the advantages of oligopeptide preparations, such as a smaller molecular weight than antibodies, which reduces the risk of immune reactions and allows for easy tissue penetration, but also have excellent cancer cell proliferation inhibitory effects and are stable in human blood, making them useful anti-cancer agents for treating cancer.

Description

[Technical field]

[0001] The present invention relates to a novel oligopeptide AQTGTGKT analogue compound, a pharmaceutical composition for preventing or treating cancer containing the same as an active ingredient, and a method for producing the same.

[0002] The present invention claims priority to Korean Patent Application No. 10-2021-0036803 filed on March 22, 2021, and PCT Application No. PCT / KR2022 / 002399 filed on February 18, 2022, the entire contents of which are incorporated herein by reference in their entirety in the specification and drawings. [Background technology]

[0003] Although the effectiveness of cancer treatment has improved with the development of early cancer diagnosis methods and the continuous development of new anti-cancer therapies, cancer is still a serious disease that ranks first or second among causes of death in Korea. Most of the anti-cancer drugs currently in use are chemotherapy drugs, which have been pointed out as problems in cancer treatment because they have various pharmacological actions depending on the type of cancer and various side effects due to toxicity.

[0004] Conventional anticancer drugs penetrate not only cancer cells but also normal cells, damaging the function and activity of normal cells, which causes side effects such as decreased bone marrow function, gastrointestinal disorders, and alopecia, and shows multidrug resistance to anticancer drugs due to long-term chemotherapy, etc. Therefore, research is being actively conducted to develop innovative anticancer drugs that can solve these serious problems of conventional anticancer drugs.

[0005] Meanwhile, antibodies targeting specific tumor antigens in tumor cells have been developed, but antibodies have problems such as the risk of immune reactions and low efficiency of tissue penetration. On the other hand, peptides have the advantage of being small in molecular weight, less likely to cause immune reactions compared to antibodies, and easy to penetrate into tissues, and peptide-based anticancer drugs targeting tumor antigens are expected to have almost no side effects such as damage to normal cells because they can act selectively on tumors. However, despite these advantages, they are only used for very limited cancers, and there are problems with them being decomposed in a short time immediately after administration to humans, making them difficult to exert their effects.

[0006] To solve the above problems, peptides may be modified by amidation, esterification, acylation, acetylation, PEGylation, cyclization, or alkylation to improve yield, increase in vivo activity, increase affinity of peptides to receptors, and delay protein degradation, but the above modifications do not guarantee that the desired effects will be improved, and there is a risk that the peptide may lose its inherent in vivo activity or cause unexpected side effects. Therefore, research into peptide modifications to maximize the desired effects while minimizing such negative effects is being actively conducted. Summary of the Invention [Problem to be solved by the invention]

[0007] The present invention has been made to solve the above-mentioned problems in the prior art, and has been completed as a result of extensive efforts to further improve the anti-cancer effect and half-life in blood of oligopeptide AQTGTGKT (alanine-glutamine-threonine-glycine-threonine-glycine-lysine-threonine). In particular, it has been confirmed that an amidated analog compound of AQTGTGKT peptide has excellent anti-cancer effect and increased half-life.

[0008] It is therefore an object of the present invention to provide a compound represented by the general formula:

[0009] [General formula] X-AQTGTGKT

[0010] In the above general formula, A is alanine, Q is glutamine, T is threonine, G is glycine, and K is lysine. X is [ka] One or more selected from the group consisting of:

[0011] Another object of the present invention is to provide a pharmaceutical composition for preventing or treating cancer, which comprises a compound represented by the above general formula as an active ingredient.

[0012] It is still another object of the present invention to provide a method for producing the compound represented by the above general formula.

[0013] However, the technical problems that the present invention aims to solve are not limited to those mentioned above, and other problems not mentioned will be clearly understood by a person having ordinary skill in the technical field to which the present invention pertains from the following description. [Means for solving the problem]

[0014] In order to achieve the above object of the present invention, the present invention provides a compound represented by the following general formula:

[0015] [General formula] X-AQTGTGKT

[0016] In the above general formula, A is alanine, Q is glutamine, T is threonine, G is glycine, and K is lysine. X is [ka] One or more selected from the group consisting of:

[0017] The present invention also provides a pharmaceutical composition for preventing or treating cancer, comprising, as an active ingredient, a compound represented by the following general formula or a pharma- ceutically acceptable salt thereof:

[0018] [General formula] X-AQTGTGKT

[0019] In the above general formula, A is alanine, Q is glutamine, T is threonine, G is glycine, and K is lysine. X is [ka] One or more selected from the group consisting of:

[0020] The present invention also provides a method for preventing or treating cancer, comprising administering to an individual in need thereof a compound represented by the general formula or a pharma- ceutical acceptable salt thereof. The compound may be administered in an effective amount.

[0021] In addition, the present invention provides a use of the compound represented by the above general formula or a pharma- ceutical acceptable salt thereof for preventing, ameliorating or treating cancer.

[0022] Furthermore, the present invention provides use of the compound represented by the above general formula or a pharma- ceutical acceptable salt thereof for producing a medicament for treating cancer.

[0023] In one embodiment of the present invention, the cancer may be a cancer selected from the group consisting of lung cancer, breast cancer, blood cancer, colon cancer, pancreatic cancer, and combinations thereof, but is not limited thereto.

[0024] In another embodiment of the present invention, the lung cancer may be, but is not limited to, non-small cell lung cancer.

[0025] In yet another embodiment of the present invention, the blood cancer may be selected from the group consisting of, but not limited to, leukemia, lymphoma, multiple myeloma, and combinations thereof.

[0026] In one embodiment of the present invention, X is [ka] and the cancer may be, but is not limited to, lung cancer.

[0027] In another embodiment of the present invention, X is [ka] and the cancer may be, but is not limited to, breast cancer.

[0028] In still another embodiment of the present invention, X is [ka] and the cancer may be, but is not limited to, a blood cancer.

[0029] In still another embodiment of the present invention, X is [ka] and the cancer may be, but is not limited to, pancreatic cancer.

[0030] In still another embodiment of the present invention, X is [ka] and the cancer may be, but is not limited to, colon cancer.

[0031] In one embodiment of the present invention, X is [ka] In this case, the compound may have a half-life in human blood of 100 minutes to 150 minutes, but is not limited thereto.

[0032] In another embodiment of the present invention, X is [ka] When the compound is one or more selected from the group consisting of, the compound may have a half-life in human blood of 45 minutes to 70 minutes, but is not limited thereto.

[0033] In yet another embodiment of the present invention, X is [ka] In this case, the compound may have a half-life in human blood of 20 minutes to 30 minutes, but is not limited thereto.

[0034] The present invention also provides a method for producing the oligopeptide X-AQTGTGKT, comprising the steps of:

[0035] (A is alanine, Q is glutamine, T is threonine, G is glycine, and K is lysine, The X is [ka] It is one selected from the group consisting of.

[0036] (1) synthesizing TG and KT, respectively; (2) synthesizing TGKT by combining the TG and KT; (3) synthesizing TGTGKT by binding TG to the N-terminus of the TGKT; (4) synthesizing QTGTGKT by binding Q to the N-terminus of the TGTGKT; and (5) synthesizing X-AQTGTGKT by binding an alanine derivative (XA) to the N-terminus of QTGTGKT. Effect of the Invention

[0037] The present invention relates to a novel oligopeptide AQTGTGKT analog compound, a pharmaceutical composition for preventing or treating cancer containing the same as an active ingredient, and a method for preparing the same, and it has been confirmed that the oligopeptide AQTGTGKT analog exhibits excellent anti-cancer effects and is stable in human blood. Therefore, the pharmaceutical composition according to the present invention has the advantages of oligopeptides, that is, a smaller molecular weight than antibodies, less risk of immune reaction, and easy penetration into tissues, and is also excellent in the effect of inhibiting the proliferation of cancer cells and exhibits the effect of being stable in human blood, so it is expected to be used as a useful anti-cancer agent for treating cancer. [Brief description of the drawings]

[0038] [Figure 1-7] Figures 1 to 7 show the results of UPLC-MS (top) and 1H NMR (bottom) for identifying the compounds according to the present invention and confirming their structures. Figure 1 shows the measurement results for 4-PhPh-AQTGTGKT, Figure 2 shows the measurement results for Ac-AQTGTGKT, Figure 3 shows the measurement results for 3-PhPh-AQTGTGKT, Figure 4 shows the measurement results for 4-MeOPh-AQTGTGKT, Figure 5 shows the measurement results for 2-PhPh-AQTGTGKT, Figure 6 shows the measurement results for Ph-AQTGTGKT, and Figure 7 shows the measurement results for Naphthyl-AQTGTGKT. [Figure 8] FIG. 8 shows the results of analyzing the cell activity suppression effect of treatment with the compound of the present invention in the lung cancer cell line H1299 by CTG assay (*p<0.05, **p<0.01, ***p<0.001; the same applies below). [Figure 9] FIG. 9 shows the results of analyzing the cell activity suppressing effect of treatment with the compound of the present invention in the lung cancer cell line H1975 by CTG assay. [Figure 10] FIG. 10 shows the results of analyzing the cell activity suppressing effect of treatment with the compound of the present invention in lung cancer (papillary adenocarcinoma) cell line H820 by MTT assay. [Figure 11a]FIG. 11a shows the results of analyzing the cell activity suppression effect of treatment with the compound of the present invention in breast cancer cell line MDA-MB-231 by CTG assay. [Figure 11b] FIG. 11b shows the results of analyzing the cell activity suppressing effect of treatment with the compound of the present invention in breast cancer cell line HCC1937 by MTT assay. [Figure 12] FIG. 12 shows the results of analyzing the cell activity suppressing effect of treatment with the compound of the present invention in blood cancer cell line Jurkat clone E6-1 by MTT assay. [Figure 13] FIG. 13 shows the results of analyzing the cell activity suppressing effect by treatment with the compound of the present invention in the pancreatic cancer cell line CFPAC-1 by MTT assay. [Figure 14] FIG. 14 shows the results of analyzing the cell activity suppressing effect by treatment with the compound of the present invention in colon cancer cell line HT29 by MTT assay. [Figure 15] FIG. 15 shows the results of analyzing the inhibition of tumor growth by treatment with the compound of the present invention after tumor formation in nude mice inoculated with lung cancer cell line H820. [Figure 16] FIG. 16 shows the results of analyzing the inhibition of tumor growth by treatment with the compound of the present invention after tumor formation in nude mice inoculated with lung cancer cell line H1975. [Figure 17] FIG. 17 shows the results of analyzing tumor growth inhibition by treatment with the compound of the present invention after tumor formation in mice inoculated with breast cancer cell line HCC1806. [Figure 18] FIG. 18 shows the results of analyzing tumor growth inhibition by treatment with the compound of the present invention after tumor volume was measured after tumor formation in mice inoculated with colon cancer cell line CT26. [Figure 19-26] 19 to 26 are diagrams showing the results of measuring the residual amount of each compound according to the present invention in human blood by time period. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0039] The present inventors have newly synthesized amidation analogs of the oligopeptide AQTGTGKT, and have confirmed that these analogs exhibit excellent anticancer effects (see Example 2) and excellent stability in blood (see Example 3), thereby completing the present invention. Thus, the present invention can provide a compound represented by the following general formula:

[0040] [General formula] X-AQTGTGKT

[0041] In the above general formula, A is alanine, Q is glutamine, T is threonine, G is glycine, K is lysine, and X is [ka] One or more selected from the group consisting of:

[0042] In another aspect, the present invention can provide a pharmaceutical composition for preventing or treating cancer, comprising a compound represented by the above general formula as an active ingredient.

[0043] In yet another aspect, the present invention can provide a method for preventing or treating cancer, comprising administering to an individual a compound represented by the general formula.

[0044] As used herein, the term "prevention" refers to any action that blocks, inhibits, or delays the symptoms of cancer by administering a composition according to the present invention.

[0045] As used herein, the term "treatment" refers to any action in which symptoms of cancer are ameliorated or beneficially altered by administration of a composition according to the present invention.

[0046] As used herein, the term "individual" refers to a subject in need of disease prevention or treatment. For example, the individual may be a mammal, including a human or non-human primate, mouse, dog, cat, horse, sheep, and cow.

[0047] The term "oligopeptide" as used herein means a linear molecule formed by linking amino acid residues together through peptide bonds. The oligopeptides of the present invention can be prepared by chemical synthesis methods known in the art (e.g., solid-phase synthesis techniques) as well as molecular and biological methods (Merrifield, J. Amer. Chem. Soc. 85:2149-54 (1963); Stewart, et al., Solid Phase Peptide Synthesis, 2nd.ed., Pierce Chem. Co.: Rockford, 111 (1984)).

[0048] The scope of the compounds according to the present invention may further include pharma- ceutically acceptable salts thereof. As used herein, the term "pharma- ceutically acceptable" refers to a compound that is suitable for use in contact with the tissues of a subject (e.g., a human) without excessive toxicity, irritation, allergic reaction, or other problems or complications, with a reasonable benefit / risk ratio, and within the bounds of sound medical judgment. The pharma- ceutically acceptable salts include, for example, acid addition salts formed with pharma- ceutical acceptable free acids, and pharma- ceutical acceptable metal salts.

[0049] The scope of the compounds according to the present invention may also include biologically functional equivalents having amino acid sequence variations that exhibit equivalent biological activity to the compounds of the present invention. Such variations in amino acid sequence are based on the relative similarity of amino acid side chain substitutions, such as hydrophobicity, hydrophilicity, charge and size. Analysis of the size, shape and type of amino acid side chain substitutions reveals that alanine and glycine have similar sizes; lysine is a positively charged residue; and glutamine and threonine are uncharged. Therefore, based on these considerations, alanine and glycine; and glutamine and threonine are biologically functional equivalents.

[0050] When making mutations, the hydropathic index of amino acids can be taken into consideration. Each amino acid is assigned a hydropathic index based on its hydrophobicity and charge as follows: isoleucine (+4.5); valine (+4.2); leucine (+3.8); phenylalanine (+2.8); cysteine ​​(+2.5); methionine (+1.9); alanine (+1.8); glycine (-0.4); threonine (-0.7); serine (-0.8); tryptophan (-0.9); tyrosine (-1.3); proline (-1.6); histidine (-3.2); glutamic acid (-3.5); glutamine (-3.5); aspartic acid (-3.5); asparagine (-3.5); lysine (-3.9); and arginine (-4.5). The hydrophobic amino acid index is very important in imparting interactive biological functions to proteins. It is a known fact that substitution with an amino acid having a similar hydrophobic index can retain similar biological activity. When introducing a mutation with reference to the hydrophobic index, substitution is preferably performed between amino acids showing a hydrophobic index difference within ±2, more preferably within ±1, and more preferably within ±0.5.

[0051] On the other hand, it is well known that substitutions between amino acids with similar hydrophilicity values ​​result in proteins with equivalent biological activity. As disclosed in U.S. Patent No. 4,554,101, the following hydrophilicity values ​​are assigned to each amino acid residue: arginine (+3.0); lysine (+3.0); aspartic acid (+3.0±1); glutamic acid (+3.0±1); serine (+0.3); asparagine (+0.2); glutamine (+0.2); glycine (0); threonine (-0.4); proline (-0.5±1); alanine (-0.5); histidine (-0.5); cysteine ​​(-1.0); methionine (-1.3); valine (-1.5); leucine (-1.8); isoleucine (-1.8); tyrosine (-2.3); phenylalanine (-2.5); tryptophan (-3.4).

[0052] When mutations are introduced with reference to the hydrophilicity value, substitutions are made between amino acids showing a difference in hydrophilicity value within ±2, more preferably within ±1, and more preferably within ±0.5.

[0053] Amino acid exchanges in proteins that do not globally alter the activity of the molecule are known in the art (H. Neurath, R.L. Hill, The Proteins, Academic Press, New York, 1979). The most commonly occurring exchanges are between the amino acid residues Ala / Ser, Val / Ile, Asp / Glu, Thr / Ser, Ala / Gly, Ala / Thr, Ser / Asn, Ala / Val, Ser / Gly, Tyr / Phe, Ala / Pro, Lys / Arg, Asp / Asn, Leu / Ile, Leu / Val, Ala / Glu, and Asp / Gly.

[0054] Considering the above-mentioned biologically equivalent activity mutations, the protein of the amino acid sequence (AQTGTGKT) of the compound of the present invention represented by the above general formula is interpreted as including a sequence showing substantial identity thereto. The substantial identity means a sequence showing at least 62.5% homology, more preferably 75% or more homology, and most preferably 87.5% or more homology when the above-mentioned sequence of the present invention is aligned with any other sequence so as to correspond as much as possible, and the aligned sequence is analyzed using an algorithm commonly used in the art. The alignment method for sequence comparison is well known in the art.

[0055] The pharmaceutical composition of the present invention is used for the prevention or treatment of cancer. The cancer for which the pharmaceutical composition of the present invention can be used may be, but is not limited to, cancer selected from the group consisting of lung cancer, breast cancer, blood cancer, colon cancer, pancreatic cancer, and combinations thereof.

[0056] In the present invention, the lung cancer may be, but is not limited to, non-small cell lung cancer. In one embodiment of the present invention, the compound according to the present invention is administered to patients with T790M mutation positive, EGFR m - The present invention may be, but is not limited to, for the treatment of lung cancer in patients and / or Osimertinib-resistant patients.

[0057] The breast cancer may be, but is not limited to, hormone receptor (HR) positive breast cancer, and may be, but is not limited to, triple negative breast cancer.

[0058] Additionally, the blood cancer may be a blood cancer selected from the group consisting of leukemia, lymphoma, multiple myeloma, and combinations thereof, but is not limited thereto.

[0059] In one embodiment of the present invention, it was confirmed that the pharmaceutical composition according to the present invention exhibits excellent anti-cancer activity against lung cancer, breast cancer, blood cancer, pancreatic cancer and colon cancer (see Example 2).

[0060] In the present invention, when the pharmaceutical composition is used for the prevention or treatment of lung cancer, it contains a compound represented by the general formula X-AQTGTGKT as an active ingredient, wherein X is [ka] The present invention is not limited to the above.

[0061] In addition, in the present invention, when the pharmaceutical composition is used for the prevention or treatment of breast cancer, it contains a compound represented by the general formula X-AQTGTGKT as an active ingredient, wherein X is [ka] The number of the at least one selected from the group consisting of, but not limited to, may be one or more selected from the group consisting of:

[0062] In addition, in the present invention, when the pharmaceutical composition is used for the prevention or treatment of blood cancer, it contains a compound represented by the general formula X-AQTGTGKT as an active ingredient, wherein X is [ka] The number of the at least one selected from the group consisting of, but not limited to, may be one or more selected from the group consisting of:

[0063] In addition, in the present invention, when the pharmaceutical composition is used for the prevention or treatment of pancreatic cancer, it contains a compound represented by the general formula X-AQTGTGKT as an active ingredient, wherein X is [ka] The number of the at least one selected from the group consisting of, but not limited to, may be one or more selected from the group consisting of:

[0064] In addition, in the present invention, when the pharmaceutical composition is used for the prevention or treatment of colon cancer, it contains a compound represented by the general formula X-AQTGTGKT as an active ingredient, wherein X is [ka] The number of the at least one selected from the group consisting of, but not limited to, may be one or more selected from the group consisting of:

[0065] In another embodiment of the present invention, it was confirmed that the compound represented by the general formula according to the present invention can exist stably in human blood and has an improved half-life compared to AQTGTGKT (see Example 3). Therefore, the results of the above embodiment show that the compound according to the present invention has improved anti-cancer effect and stability.

[0066] In the compound of the present invention represented by the general formula X-AQTGTGKT, [ka] In this case, the half-life of the compound in human blood may be, but is not limited to, 20 minutes or more. The half-life may be, for example, 20 minutes to 30 minutes, 21 minutes to 30 minutes, 22 minutes to 30 minutes, 23 minutes to 30 minutes, 24 minutes to 30 minutes, 25 minutes to 30 minutes, 26 minutes to 30 minutes, 27 minutes to 30 minutes, 28 minutes to 30 minutes, 29 minutes to 30 minutes, 20 minutes to 29 minutes, 21 minutes to 29 minutes, 21 minutes to 28 minutes, 21 minutes to 27 minutes, 21 minutes to 26 minutes, 21 minutes to 25 minutes, 21 minutes to 24 minutes, 21 minutes to 23 minutes, 21 minutes to 2 ... It may be 2 minutes to 30 minutes, 22 minutes to 28 minutes, 22 minutes to 26 minutes, 22 minutes to 24 minutes, 23 minutes to 30 minutes, 23 minutes to 28 minutes, 23 minutes to 26 minutes, 23 minutes to 24 minutes, 24 minutes to 30 minutes, 24 minutes to 27 minutes, 25 minutes to 30 minutes, 25 minutes to 27 minutes, 26 minutes to 30 minutes, 26 minutes to 28 minutes, 27 minutes to 30 minutes, 27 minutes to 29 minutes, 28 minutes to 30 minutes, 28 minutes to 29 minutes, or 29 minutes to 30 minutes, etc. Furthermore, the half-life may be increased by 1900% to 2900% compared to the half-life of AQTGTGKT, but is not limited thereto. The increase ratio may be, for example, 1900% to 2700%, 2000% to 2700%, 2100% to 2700%, 2200% to 2700%, 2300% to 2700%, 2400% to 2700%, 2500% to 2700%, 2600% to 2700%, 1900% to 2600%, 2000% to 2600%, 2100% to 2600%, 2100% to 2500%, 2100% to It may be 2400%, 2100% to 2300%, 2100% to 2200%, 2200% to 2700%, 2200% to 2600%, 2200% to 2500%, 2200% to 2400%, 2200% to 2300%, 2300% to 2700%, 2300% to 2500%, 2400% to 2700%, 2400% to 2600%, or 2500% to 2700%, etc.

[0067] In addition, in the compound of the present invention represented by the general formula X-AQTGTGKT, [ka] When the compound is one or more selected from the group consisting of, the half-life of the compound in human blood may be 45 minutes or more, but is not limited thereto. The half-life may be, for example, 45 minutes to 70 minutes, 46 minutes to 70 minutes, 47 minutes to 70 minutes, 48 ​​minutes to 70 minutes, 49 minutes to 70 minutes, 50 minutes to 70 minutes, 51 minutes to 70 minutes, 53 minutes to 70 minutes, 55 minutes to 70 minutes, 57 minutes to 70 minutes, 59 minutes to 70 minutes, 65 minutes to 70 minutes, 45 minutes to 65 minutes, 50 minutes to 65 minutes, 55 minutes to 65 minutes, 60 minutes to 65 minutes, 45 minutes to 60 minutes, 50 minutes to 60 minutes, 55 minutes to 60 minutes, 45 minutes to 55 minutes, 50 minutes to 55 minutes, or 60 minutes to 70 minutes. Furthermore, the half-life may be increased by 4400% to 6900% compared to the half-life of AQTGTGKT, but is not limited thereto. The increase ratio may be, for example, 4400% to 6900%, 4500% to 6900%, 4600% to 6900%, 4700% to 6900%, 4800% to 6900%, 4900% to 6900%, 5000% to 6900%, 5100% to 6900%, 5200% to 6900%, 5300% to 6900%, 5400% to 6900%, 5500% to 6900%, 5600% to 6900%, 5700% to 6900%, 5800% to 6900%, 5900% to 6900%, 6000% to 6900%, 6100% to 6900%, 6200% to 6900%, 6300% to 6900%, 6400% to 6900%, 6500% to 6900%, 6600% to 6900%, 6700% to 6900%, 6800% to 6900%, 6900% to 6900%, 7000% to 7000%, 7100% to 7000%, 7200% to 7000%, 7300% to 7000%, 7400% to 7000%, 7500% to 7000%, 7600% to 7000%, 7700% to 7000%, 7800% to 7000%, 79 0%~6900%, 5600%~6900%, 5700%~6900%, 5800%~6900%, 5900%~6900%, 6000%~6900%, 6100%~6900%, 6200%~6900%, 6300%~6900%, 6400%~6900%, 6500%~6900%, 6600%~6900%, 6700%~ 6900%, 6800%~6900%, 4400%~6500%, 4500%~6500%, 4700%~6500%, 5000%~6500%, 5200%~6500%, 5500%~6500%, 5700%~6500%, 6000%~6500%, 6300%~6500%, 4400%~6000%, 4600%~600 It may be 0%, 4800% to 6000%, 5000% to 6000%, 5500% to 6000%, 4400% to 5500%, 4700% to 5500%, 5000% to 5500%, 5200% to 5500%, 4400% to 5000%, 4800% to 5000%, 4400% to 4700%, or 4500% to 4700%, etc.

[0068] In addition, in the compound of the present invention represented by the general formula X-AQTGTGKT,

change

[0069] Meanwhile, the pharmaceutical composition according to the present invention may further contain suitable carriers, excipients and / or diluents that are generally used for preparing pharmaceutical compositions, in addition to the active ingredient, and may be formulated into oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, external preparations, suppositories, and sterile injection solutions by a conventional method.

[0070] The carrier, excipient and diluent that can be contained in the composition include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil.When formulating the composition, it can be prepared using diluents or excipients such as commonly used fillers, extenders, binders, wetting agents, disintegrants, and surfactants.

[0071] The pharmaceutical composition according to the present invention is administered in a pharma- ceutical effective amount. In the present invention, the term "pharma-ceutical 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 can be determined by factors including the type and severity of the patient's disease, the activity of the drug, sensitivity to the drug, administration time, administration route and excretion rate, treatment period, drugs used concomitantly, and other factors well known in the medical field. A preferred dosage can be selected according to the condition and weight of the individual, the degree of the disease, the drug form, the administration route and period. As a specific example, the pharmaceutical composition can be administered in an amount of 0.001 to 1000 mg / kg, 0.01 to 100 mg / kg, 0.01 to 10 mg / kg, 0.1 to 10 mg / kg, or 0.1 to 1 mg / kg once or several times a day.

[0072] It is important to administer an amount that can obtain maximum effect at a minimum amount without side effects, taking into consideration all the above factors, and this can be determined by those skilled in the art. Specifically, the effective amount of the pharmaceutical composition according to the present invention may vary depending on the patient's age, sex, condition, weight, absorption degree, inactivation rate and excretion rate of the active ingredient in the body, type of disease, and drugs used in combination.

[0073] The pharmaceutical composition of the present invention can be administered to an individual by various routes. For example, it can be administered by oral administration, intranasal administration, transbronchial administration, intraarterial injection, intravenous injection, subcutaneous injection, intramuscular injection, or intraperitoneal injection. The daily dose can be administered once or several times a day.

[0074] In another aspect, the present invention provides a method for producing oligopeptide X-AQTGTGKT, comprising the steps of:

[0075] (A is alanine, Q is glutamine, T is threonine, G is glycine, and K is lysine, The X is [ka] It is one selected from the group consisting of. (1) synthesizing TG and KT, respectively; (2) synthesizing TGKT by combining the TG and KT; (3) synthesizing TGTGKT by binding TG to the N-terminus of the TGKT; (4) synthesizing QTGTGKT by binding Q to the N-terminus of the TGTGKT; and (5) synthesizing X-AQTGTGKT by binding an alanine derivative (XA) to the N-terminus of QTGTGKT. EXAMPLES

[0076] The terms and words used in this specification and claims should not be interpreted in a limited manner to their ordinary or dictionary meanings, but should be interpreted in a manner that corresponds to the technical idea of ​​the present invention based on the principle that the inventor can appropriately define the concept of the term in order to best describe his / her invention.

[0077] In the following, preferred examples are presented to aid in understanding the present invention. However, the following examples are provided only to facilitate understanding of the present invention, and the contents of the present invention are not limited to the following examples.

[0078] [Example] Experimental Method 1.CTG (CellTiter-Glo Luminescent) analysis After the AQTGTGKT analog compound according to the present invention was applied to a cancer cell line, the degree of cell proliferation was measured by CTG analysis. 3After seeding 100 μl of cells and culturing for 24 hours, the seven AQTGTGKT analogs according to the present invention were transfected. After 48 hours, the same amount of CellTiter-Glo (Promega Co., USA) reagent was mixed with the cell culture medium and reacted for 2 minutes on an orbital shaker. After reacting for 10 minutes at room temperature, the luminescence signal was measured using a luminometer (GloMax, Promega).

[0079] 2.MTT (tetrazolium) analysis After the AQTGTGKT analog compound according to the present invention was applied to a cancer cell line, the degree of cell proliferation was measured by MTT assay. 3 After 24 hours of incubation, 7 types of AQTGTGKT analogs according to the present invention were introduced into the wells. After 72 hours, 10 μl of CellTiter-96 (Promega Co., USA) reagent was added to each well and incubated at 5% CO2 and 37°C. After 3 hours, the cells were measured using a spectrophotometer (SPECTROstar Nano The absorbance was measured at 490 nm using a 100% PBS (BMG).

[0080] 3. Lung cancer animal model 4 × 10 H1975 cells 6 The mice were inoculated once by subcutaneous injection into the flank of each mouse at a concentration of 150 μl of 100 cells / mouse. After completion of inoculation, the volume of the formed tumor mass was 70-130 mm 3 After confirming that the rats had reached the target concentration, the rats were randomly divided into groups. The test substance was administered via the tail vein five times at three-day intervals and five times at two-day intervals. 5 × 10 H820 cells 6Mice were inoculated once subcutaneously into the flanks of the mice with 200μl of Matrigel at a ratio of 1:1 at a concentration of 100 cells / mouse. After inoculation, it was confirmed that the volume of the tumor mass formed reached 70-130mm3, and then randomized into groups. The test substance was administered via the tail vein 14 times daily.

[0081] 4. Breast Cancer Animal Models Breast cancer cell line HCC1806 cells were cultured at 5 × 10 6 The mice were inoculated once subcutaneously into the flanks of the mice at a concentration of 1:1 cells / mouse with Matrigel (200 μl each). After inoculation, the tumor volume was 70-130 mm. 3 After confirming that the mice had reached the target mass, random group separation was performed. The test substance was administered into the tail vein at a concentration of 10mpk 7 times at 2-day intervals, with the administration start date set as the 1st. The tumor volume was measured twice a week, and the long and short axis diameters were measured to calculate the volume, and the absolute weight of each individual was measured at the same time.

[0082] 5. Colon Cancer Animal Model CT26 cell line in which CAGE gene expression was induced was cultured at 1 × 10 6 Mice were inoculated once by subcutaneous injection into the flank of 200μl of Matrigel at a concentration of 1:1 cells / mouse. After completion of inoculation, it was confirmed that the volume of the formed tumor reached 70mm3, and random group division was performed. The test substance was administered into the tail vein at a concentration of 10mpk 7 times at 2-day intervals, with the first day of administration set as the 1st. The tumor volume was measured twice a week, and the long and short axis diameters were measured to calculate the volume, and the absolute weight of each individual was measured at the same time.

[0083] Example 1: Preparation of AQTGTGKT analog 1.1. Reactions in general All reactions were carried out with commercially available materials and reagents without further processing unless otherwise stated. Reactions were monitored by thin layer chromatography (TLC) on silica gel plates (Keiselgel 60 F254, Merck) and / or ultra-performance liquid chromatography (UPLC). Visualization of spots on the TLC plates was achieved by UV light and by staining the TLC plates with potassium permanganate and / or ninhydrin and charring with a heat gun. All products were 1 Characterization was performed using 1 H NMR and / or UPLC-MS.

[0084] 1.2.Synthesis of Boc / OBn-TG First, TG whose functional group was protected with benzyl was synthesized according to the following reaction scheme 1. Hereinafter, in each reaction scheme, the compound will be referred to as compound n by the Arabic numeral (n) shown below.

[0085] [Reaction Scheme 1] [ka]

[0086] Specifically, BocThr(OBn)OH (compound 1; 25.0 g, 80.8 mmol, 1.0 equiv.) and NOSu (9.77 g, 84.8 mmol, 1.05 equiv.) were dissolved in dichloromethane (150 mL). The mixture was cooled to 0° C. and placed under an inert atmosphere. To the mixture was then added 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (16.3 g, 84.8 mmol, 1.05 equiv.). The mixture was allowed to warm to room temperature and stirred for 20 h. The mixture was then washed with NH4Cl (saturated aqueous) and the phases were separated. The organic layer was dried over MgSO4 and concentrated under reduced pressure to give the product compound 2 as a pale yellow oil (35.7 g, >100% yield, assuming quantitative yield).

[0087] The compound 2, BocThr(OBn)OSu (32.8 g, 80.8 mmol, 1.0 equiv.), was dissolved in 1,4-dioxane (200 mL) and a solution of glycine sodium salt hydrate in distilled water (100 mL) was added in one portion. After stirring at room temperature for 6 h, the mixture was fractionated into ethyl acetate and citric acid (saturated aqueous). The organic layer was dried over MgSO4, filtered, and concentrated under reduced pressure. The crude material was purified on a C18 (400 g) column with 30-70% acetonitrile (0.1% formic acid) in water (0.1% formic acid) eluent. The desired fractions were combined and fractionated into ethyl acetate and NaHCO3 (saturated aqueous). The organic layer was dried over MgSO4, filtered, and concentrated under reduced pressure to obtain the product, compound 3, as a pale yellow gum (21.9 g, 74% yield).

[0088] 1.3.Synthesis of CBz / OBn / CO2Bn-KT KT, in which the OH functional group was protected with benzyl, was synthesized according to the following reaction scheme 2.

[0089] [Reaction Scheme 2] [ka]

[0090] Specifically, BocLys(CBz)OH (compound 4; 27.0 g, 70.9 mmol, 1.0 equiv.) and NOSu (9.80 g, 85.1 mmol, 1.2 equiv.) were dissolved in dichloromethane (128 mL). The mixture was cooled to 0° C. and placed under an inert atmosphere. To the mixture was then added 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (16.3 g, 85.1 mmol, 1.05 equiv.). The mixture was allowed to warm to room temperature and stirred for 20 h. The mixture was then washed with NH4Cl (saturated aqueous) and the phases were separated. The organic layer was dried over MgSO4 and concentrated under reduced pressure to give the product, compound 5, as a pale yellow oil (36.7 g, >100% yield, assuming quantitative yield).

[0091] Next, the compound 5 (BocLys(Cbz)OSu; 36.7 g, 70.9 mmol, 1.0 eq.) and Thr(OBn)OBn.HCl (25.0 g, 74.4 mmol, 1.05 eq.) were dissolved in 1,4-dioxane (477 mL) at room temperature. A solution of NaHCO3 (6.85 g, 81.5 mmol, 1.15 eq.) in distilled water (326 mL) was added to the solution. The resulting mixture was then stirred at room temperature for 20 hours. The reaction mixture was diluted with ethyl acetate and washed with 10% citric acid (aqueous) and brine. The organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain 55.9 g (>100% yield, assuming quantitative yield) of a yellow oily solid, compound 6, as the product.

[0092] Finally, compound 6 (BocLys(Cbz)Thr(OBn)OBn; 55.9 g, 70.9 mmol, 1.00 eq.) was dissolved in 1,4-dioxane (360 mL) and 4N HCl in 1,4-dioxane (177 mL) was added. The mixture was stirred at room temperature overnight. Then, a saturated aqueous solution of NaHCO3 was added until the pH value reached 8. The solution was extracted with ethyl acetate, and the resulting organic solution was dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain compound 7 as a yellow oily solid (38.7 g, 97% yield).

[0093] 1.4.Synthesis of CBz / OBn / OBn / CO2Bn-TGKT TG and KT synthesized in 1.2. and 1.3. above were combined according to the following reaction formula 3 to synthesize TGKT.

[0094] [Reaction Scheme 3] [ka]

[0095] More specifically, to a solution of BocThr(OBn)GlyOH (compound 3; 5.61 g, 15.3 mmol, 1.00 equiv.) and compound 8 (Lys(Cbz)Thr(OBn)OBn; 10.0 g, 15.3 mmol, 1.0 equiv.) in dichloromethane (50 mL), N,N-diisopropylethylamine (5.90 mL, 33.7 mmol, 2.2 equiv.) was added. The mixture was stirred at room temperature under an inert atmosphere, and HATU (7.00 g, 18.4 mmol, 1.20 equiv.) was added. The resulting mixture was stirred for 2 h, then washed with NH4Cl (saturated aqueous) and then with NaHCO3 (saturated aqueous). The organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure to give compound 9 as a pale orange oily solid (25.0 g, >100% yield, assuming quantitative yield) as the product.

[0096] The obtained BocThr(OBn)GlyLys(Cbz)Thr(OBn)OBn (compound 9; 13.9 g, 15.3 mmol, 1.0 equiv. taken from the previous step) was dissolved in 1,4-dioxane (150 mL) at room temperature under nitrogen. To this solution was added 4N HCl in 1,4-dioxane (20 mL). The mixture was stirred at room temperature for 20 hours. The mixture was concentrated under reduced pressure and purified on a C18 (400 g) column using 20% ​​acetonitrile (0.1% formic acid) in water (0.1% formic acid) eluent. The desired fractions were combined and lyophilized. The resulting powder was dissolved in NaHCO3 (saturated aqueous) and dichloromethane and stirred for 15 minutes. The layers were separated and the organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure to give the product, compound 10, as a colorless gum (10.9 g, 88% yield).

[0097] 1.5. Synthesis of CBz / OBn / OBn / OBn / CO2Bn-TGTGKT TG (compound 3) and TGKT (compound 10) synthesized in 1.2. and 1.4. above were combined according to the following reaction formula 4 to synthesize benzyl-protected TGTGKT.

[0098] [Reaction Scheme 4] [ka]

[0099] More specifically, to a solution of BocThr(OBn)GlyOH (compound 3; 5.10 g, 14.0 mmol, 1.05 equiv.) and BocThr(OBn)GlyLys(Cbz)Thr(OBn)OBn (compound 10; 10.8 g, 13.3 mmol, 1.0 equiv.) in dichloromethane (100 mL), N,N-diisopropylethylamine (5.10 mL, 29.3 mmol, 2.2 equiv.) was added. The mixture was stirred at room temperature under an inert atmosphere, and HATU (5.60 g, 14.7 mmol, 1.1 equiv.) was added. The resulting mixture was stirred for 2 h and washed with NH4Cl (saturated aqueous) and NaHCO3 (saturated aqueous). The organic layer was concentrated under reduced pressure to give compound 11 as a pale yellow gum (21.4 g, >100% yield, assuming quantitative yield) as the product.

[0100] The obtained compound 11 (BocThr(OBn)GlyThr(OBn)GlyLys(Cbz)Thr(OBn)OBn; 15.4 g, 13.3 mmol, 1.00 equiv. taken from the previous step) was then dissolved in 1,4-dioxane (150 mL) at room temperature under nitrogen. To this solution was added 4N HCl in 1,4-dioxane (50 mL) and the mixture was stirred at room temperature for 5 h. The mixture was concentrated under reduced pressure and purified on a C18 (120 g) column using 20% ​​acetonitrile (0.1% formic acid) in water (0.1% formic acid) eluent. The desired fractions were combined and concentrated to half the volume, then fractionated between NaHCO3 (saturated aqueous) and ethyl acetate. The layers were separated and the organic layer was dried over Na2SO4, filtered and concentrated under reduced pressure to give the product compound 12 as an off-white solid (14.6 g, >100% yield, assuming quantitative yield).

[0101] Synthesis of CBz / OBn / OBn / OBn / CO2Bn-QTGTGKT Compound 12 (TGTGKT) synthesized in 1.5. above was further bonded with Q according to the following reaction formula 5 to synthesize compound 14 (QTGTGKT).

[0102] [Reaction Scheme 5] [ka]

[0103] More specifically, to a solution of Thr(OBn)GlyThr(OBn)GlyLys(Cbz)Thr(OBn)OBn (compound 12; 12.7 g, 12.0 mmol, 1.0 equiv.) and BocGlnOH (3.25 g, 13.2 mmol, 1.1 equiv.) in ethyl acetate (150 mL) and N,N-dimethylformamide (25 mL) was added N,N-diisopropylethylamine (4.60 mL, 26.4 mmol, 2.2 equiv.). The mixture was stirred at room temperature under an inert atmosphere and HATU (5.47 g, 14.4 mmol, 1.20 equiv.) was added. The resulting mixture was stirred for 1 h and then washed with NH4Cl (saturated aqueous). The organic layer was further extracted with dichloromethane. The organic layers were then combined and concentrated under reduced pressure. The crude material was purified on a 400 g C18 column using a gradient of 20-100% acetonitrile (0.1% formic acid) in water (0.1% formic acid). The desired fractions were combined and then fractionated between ethyl acetate and NaHCO3 (saturated aqueous) solutions. The organic layer was concentrated and the residue was removed by a lyophilization step. The combined fractions yielded a total of 12.9 g (89% overall yield) of compound 13.

[0104] The obtained compound 13 (BocGlnThr(OBn)GlyThr(OBn)GlyLys(Cbz)Thr(OBn)OBn; 7.00 g, 5.40 mmol, 1.00 equiv.) was dissolved in 1,4-dioxane (150 mL) at room temperature under nitrogen. 4N HCl in 1,4-dioxane (43.5 mL) was added to the solution. The mixture was stirred at room temperature for 20 hours. The mixture was concentrated under reduced pressure and lyophilized using a water / acetonitrile (2 / 1) solution. Finally, compound 14 was isolated as a pale yellow powder (6.36 g, 96% yield).

[0105] 1.7. Synthesis of AQTGTGKT analogues The remaining six analogs, excluding 4-PhPh-AQTGTGKT, were synthesized by combining compound 14, the final product of reaction scheme 5, and compound 17n, the product of reaction scheme 6, according to reaction scheme 7.

[0106] [Reaction Scheme 6] [ka]

[0107] [Reaction Scheme 7] [ka]

[0108] Through Reaction Scheme 7, 2.9 mg of 3-PhPh-AQTGTGKT with a purity of 90% or more, 7.0 mg of 4-MeOPh-AQTGTGKT with a purity of 89%, 22.7 mg of 2-PhPh-AQTGTGKT with a purity of 95% or more, 23.2 mg of Ph-AQTGTGKT with a purity of 90% or more, and 23.2 mg of Naphthyl-AQTGTGKT with a purity of 85% were finally obtained.

[0109] Each analog synthesis process will be specifically described below.

[0110] Synthesis of 1.7.1.3-PhPh-AQTGTGKT 3-PhPh-AQTGTGKT (compound 19-1) was synthesized by reacting compound 17-1 obtained according to the following reaction formula 8 with compound 14 according to reaction formula 9 to synthesize the final target compound 3-PhPh-AQTGTGKT.

[0111] [Reaction Scheme 8] [ka]

[0112] More specifically, H-Ala-OBzl.HCl (388 mg, 1.80 mmol, 1.2 equiv) was suspended in ethyl acetate (10 mL) and N,N-diisopropylethylamine (653 μL, 3.75 mmol, 2.5 equiv) was added. After stirring at room temperature for 5 min, HATU (855 mg, 2.25 mmol, 1.5 equiv) and [1,1'-biphenyl]-3-carboxylic acid (297 mg, 1.50 mmol, 1 equiv) were added and the mixture was stirred at room temperature for 2 h. The reaction mixture was diluted with ethyl acetate and then washed with NH4Cl (saturated aqueous), NaHCO3 (saturated aqueous) and brine. The collected organics were dried (Na2SO4), filtered and concentrated under reduced pressure. The residue was purified on a 25 g column with a gradient of 2-40% ethyl acetate in heptane to give compound 16-1 as a colorless solid (493 mg, 91% yield).

[0113] Then, 10% Pd / C (49 mg) wetted with a minimum amount of water was added to a solution of compound 16-1 (3-PhPh-AlaOBn; 493 mg, 1.37 mmol) in methanol (30 mL). The mixture was stirred under a hydrogen atmosphere (balloon) for 2 h. The mixture was filtered through a pad of Celite and washed with methanol and ethyl acetate. The filtrate was concentrated under reduced pressure to give compound 17-1 as a colorless foam (337 mg, 91% yield), which was reacted with compound 14 according to Scheme 9 below.

[0114] [Reaction Scheme 9] [ka]

[0115] More specifically, HATU (106 mg, 0.278 mmol, 1.1 equiv) was added to a suspension of GlnThr(OBn)GlyThr(OBn)GlyLys(Cbz)Thr(OBn)OBn (compound 14; 300 mg, 0.253 mmol, 1 equiv) and 3-PhPh-AlaOH (compound 17-1; 68.0 mg, 0.253 mmol, 1 equiv) in N,N-diisopropylethylamine (97.0 μL, 0.556 mmol, 2.2 equiv) and dichloromethane (20 mL). The mixture was stirred at room temperature for 2 h, and the reaction mixture was washed with NaHCO3 (saturated aqueous). The organic matter was concentrated under reduced pressure, and the residue was purified on a C18 column with a gradient of 40-100% acetonitrile (0.1% formic acid) in water (0.1% formic acid) (60 g) and lyophilized to obtain compound 18-1 as a colorless solid (140 mg, 38% yield).

[0116] Then, 10% Pd / C (85.0 mg) was added to a solution of 3-PhPh-AlaGlnThr(OBn)GlyThr(OBn)GlyLys(Cbz)Thr(OBn)OBn (compound 18-1; 85.0 mg, 59.1 μmol) in 2M hydrochloric acid (aqueous, 0.5 mL) and 2-propanol (10 mL). After stirring the mixture under hydrogen atmosphere (balloon) for 4.5 hours, the mixture was filtered through a 0.45 μm syringe filter. The obtained filtrate was concentrated under reduced pressure, and the residue was lyophilized. The material was then purified on a 60 g C18 column using 5-50% acetonitrile (0.1% formic acid) in water (0.1% formic acid) and lyophilized to obtain the target compound 19-1 as a colorless solid (2.9 mg, 5% yield).

[0117] Compound 19-1 (3-PhPh-AQTGTGKT) 1 H NMR data was measured as follows: 1H NMR (400MHz; D2O): δ=8.01-7.99(m, 1H), 7.86-7.82(m, 1H), 7.75-7.66(m, 3H), 7.55(t, J 7.7Hz, 1H), 7.49(t, J 7.5Hz, 2H), 7.43-7.38(m, 1H), 4.48-4.24(m, 5H), 4.23-4.12(m, 3H), 4.09(d, J 3.8Hz, 1H), 4.00-3.96(m, 2H), 3.88(s, 2H), 2.90(t, J 7.4Hz, 2H), 2.35(t, J 7.5Hz, 2H), 2.15-2.06(m, 1H), 2.03-1.91(m, 1H), 1.84-1.72(m, 1H), 1.70-1.52(m, 3H), 1.45(d, J 7.2Hz, 3H), 1.40-1.24(m, 2H), 1.15-1.05(m, 9H), 16 exchangeable protons not visible.

[0118] 1.7.2.Synthesis of 4-MeOPh-AQTGTGKT 4-MeOPh-AQTGTGKT (compound 19-2) was synthesized by reacting compound 17-2 obtained according to the following reaction formula 10 with compound 14 according to reaction formula 11 to synthesize the final target compound 4-MeOPh-AQTGTGKT.

[0119] [Reaction Scheme 10] [ka]

[0120] More specifically, H-Ala-OBzl.HCl (425 mg, 1.97 mmol, 1.2 equiv) was suspended in ethyl acetate (10 mL) and N,N-diisopropylethylamine (715 μL, 4.11 mmol, 2.5 equiv) was added. After stirring at room temperature for 5 min, HATU (937 mg, 2.46 mmol, 1.5 equiv) and 4-methoxybenzoic acid (250 mg, 1.64 mmol, 1 equiv) were added and the mixture was stirred at room temperature for 2 h. The reaction mixture was diluted with ethyl acetate and then washed with NH4Cl (saturated aqueous), NaHCO3 (saturated aqueous) and brine. The organics were dried (Na2SO4), filtered and concentrated under reduced pressure. The residue was purified on a 25 g column with a 15-50% ethyl acetate gradient in heptane to give compound 16-2 as a colorless solid (360 mg, 70% yield).

[0121] Next, 10% Pd / C (18 mg) wetted with a minimum amount of water was added to a solution of 4-OMePh-AlaOBn (compound 16-2; 180 mg, 0.574 mmol) in methanol (15 mL). The mixture was stirred under a hydrogen atmosphere (balloon) for 110 hours. The mixture was then filtered through a Celite pad and washed with methanol. The filtrate was concentrated under reduced pressure to obtain compound 17-2 as a colorless oil (128 mg, 100% yield), which was reacted with compound 14 according to the following reaction scheme 11.

[0122] [Reaction Scheme 11] [ka]

[0123] More specifically, HATU (74.5 mg, 0.196 mmol, 1.2 equiv) was added to a suspension of GlnThr(OBn)GlyThr(OBn)GlyLys(Cbz)Thr(OBn)OBn (compound 14; 200 mg, 0.164 mmol, 1 equiv) and 4-OMePh-AlaOH (compound 17-2; 36.6 mg, 0.164 mmol, 1 equiv) in N,N-diisopropylethylamine (63.0 μL, 0.360 mmol, 2.2 equiv) and dichloromethane (20 mL). The mixture was stirred at room temperature for 64 hours, and the reaction mixture was diluted with methanol and then washed with NH4Cl (saturated aqueous), NaHCO3 (saturated aqueous) and water. The organic matter was concentrated under reduced pressure, and the residue was purified on a C18 column with a gradient of 50-95% acetonitrile (0.1% formic acid) in water (0.1% formic acid) (60 g) and lyophilized to obtain compound 18-2 as a colorless solid (113 mg, 50% yield).

[0124] Then, 10% Pd / C (100 mg) was added to a solution of 4-OMePh-AlaGlnThr(OBn)GlyThr(OBn)GlyLys(Cbz)Thr(OBn)OBn (compound 18-2; 108 mg, 77.6 μmol) in 2 M hydrochloric acid (aqueous, 1.0 mL) and 2-propanol (20 mL). The mixture was stirred under a hydrogen atmosphere (balloon) for 18 hours. The mixture was filtered through a 0.45 μm syringe filter. The obtained filtrate was concentrated under reduced pressure, and the residue was dissolved in water and then lyophilized. The dried material was purified on a 30 g C18 column using a 5-30% acetonitrile (0.1% formic acid) gradient in water (0.1% formic acid) and lyophilized to obtain compound 19-2 as a colorless solid (7.0 mg, 10% yield).

[0125] Compound 19-2 (4-MeOPh-AQTGTGKT) 1 H NMR data was measured as follows: 1H NMR (400MHz; D2O): δ=7.76-7.71(m, 2H), 7.02-6.98(m, 2H), 4.42-4.28(m, 5H), 4.24-4.13(m, 3H), 4.09(d, J 3.9Hz, 1H), 4.01-3.96(m, 2H), 3.89(s, 2H), 3.81(s, 3H), 2.91(t, J 7.4Hz, 2H), 2.34(t, J 7.6Hz, 2H), 2.14-2.06(m, 1H), 2.00-1.91(m, 1H), 1.85-1.75(m, 1H), 1.71-1.54(m, 3H), 1.42(d, J 7.2Hz, 3H), 1.40-1.25(m, 3H), 1.16-1.07(m, 8H), 16 exchangeable protons not visible.

[0126] Synthesis of 1.7.3.2-PhPh-AQTGTGKT 2-PhPh-AQTGTGKT (compound 19-3) was synthesized by reacting compound 17-3 obtained according to the following reaction formula 12 with compound 14 according to reaction formula 13 to synthesize the final target compound 2-PhPh-AQTGTGKT.

[0127] [Reaction scheme 12] [ka]

[0128] More specifically, H-Ala-OBzl.HCl (388 mg, 1.80 mmol, 1.2 equiv) was suspended in ethyl acetate (10 mL) and N,N-diisopropylethylamine (653 μL, 3.75 mmol, 2.5 equiv) was added. After stirring at room temperature for 5 min, HATU (855 mg, 2.25 mmol, 1.5 equiv) and [1,1'-biphenyl]-2-carboxylic acid (297 mg, 1.50 mmol, 1 equiv) were added and the mixture was stirred at room temperature for 2 h. The reaction mixture was diluted with ethyl acetate and washed with NH4Cl (saturated aqueous), NaHCO3 (saturated aqueous) and brine. The organics were dried (Na2SO4), filtered and concentrated under reduced pressure. The residue was purified on a 25 g column with a gradient of 2-40% ethyl acetate in heptane to give compound 16-3 as a colorless oil (416 mg, 77% yield).

[0129] Next, 10% Pd / C (42 mg) wetted with a minimum amount of water was added to a solution of 2-PhPh-AlaOBn (compound 16-3; 416 mg, 1.16 mmol) in methanol (20 mL). The mixture was stirred under a hydrogen atmosphere (balloon) for 18 hours. The mixture was then filtered through a Celite pad and washed with methanol. The filtrate was concentrated under reduced pressure to obtain compound 17-3 as a colorless oil (308 mg, 99% yield), which was reacted with compound 14 according to the following reaction scheme 13.

[0130] [Reaction Scheme 13] [ka]

[0131] More specifically, HATU (74.5 mg, 0.196 mmol, 1.2 equiv) was added to a suspension of GlnThr(OBn)GlyThr(OBn)GlyLys(Cbz)Thr(OBn)OBn (compound 14; 200 mg, 0.164 mmol, 1 equiv) and 2-PhPh-AlaOH (compound 17-3; 44.2 mg, 0.164 mmol, 1 equiv) in N,N-diisopropylethylamine (63.0 μL, 0.360 mmol, 2.2 equiv) and dichloromethane (20 mL). The mixture was stirred at room temperature for 64 hours, and the reaction mixture was diluted with methanol and then washed with NH4Cl (saturated aqueous), NaHCO3 (saturated aqueous) and water. The organic layer was concentrated under reduced pressure, and the residue was purified on a C18 column with a gradient of 50-95% acetonitrile (0.1% formic acid) in water (0.1% formic acid) (60 g) and lyophilized to obtain compound 18-3 as a colorless solid (115 mg, 49% yield).

[0132] Then, 10% Pd / C (100 mg) was added to a solution of 2-PhPh-AlaGlnThr(OBn)GlyThr(OBn)GlyLys(Cbz)Thr(OBn)OBn (compound 18-3; 110 mg, 76.5 μmol) in 2 M hydrochloric acid (aqueous, 1.0 mL) and 2-propanol (20 mL). The mixture was stirred under a hydrogen atmosphere (balloon) for 18 hours. The mixture was filtered through a 0.45 μm syringe filter. The obtained filtrate was concentrated under reduced pressure, and the residue was dissolved in water and then lyophilized. The dried material was purified on a C18 column with a 5-50% acetonitrile (0.1% formic acid) gradient in water (0.1% formic acid) for 30 g, and lyophilized to obtain compound 19-3 as a colorless solid (22.7 mg, 31% yield).

[0133] Compound 19-3 (2-PhPh-AQTGTGKT) 1 H NMR data was measured as follows: 1H NMR (400MHz; D2O): δ=7.56-7.48(m, 2H), 7.44-7.33(m, 7H), 4.38-4.26(m, 4H), 4.25-4.13(m, 4H), 4.08(d, J 3.9Hz, 1H), 4.00-3.96(m, 2H), 3.89(s, 2H), 2.91(t, J 7.5Hz, 2H), 2.25(t, J 7.5Hz, 2H), 2.09-2.01(m, 1H), 1.93-1.77(m, 2H), 1.72-1.56(m, 3H), 1.41-1.29(m, 2H), 1.18(d, J 7.2Hz, 3H), 1.12(d, J 5.6Hz, 6H), 1.08(d, J 6.4Hz, 3H), 16 exchangeable protons not visible.

[0134] Synthesis of Ph-AQTGTGKT Ph-AQTGTGKT (compound 19-4) was synthesized by reacting compound 17-4 obtained according to the following reaction formula 14 with compound 14 according to reaction formula 15 to synthesize the final target compound Ph-AQTGTGKT.

[0135] [Reaction Scheme 14] [ka]

[0136] More specifically, H-Ala-OBzl.HCl (388 mg, 1.80 mmol, 1.2 equiv) was suspended in ethyl acetate (10 mL) and N,N-diisopropylethylamine (653 μL, 3.75 mmol, 2.5 equiv) was added. After stirring at room temperature for 5 minutes, HATU (855 mg, 2.25 mmol, 1.5 equiv) and benzoic acid (183 mg, 1.50 mmol, 1 equiv) were added and the mixture was stirred at room temperature for 18 hours. The reaction mixture was diluted with ethyl acetate and washed with NH4Cl (saturated aqueous), NaHCO3 (saturated aqueous) and brine. The organics were dried (Na2SO4), filtered and concentrated under reduced pressure. The residue was purified on a 25 g column with a 2-50% ethyl acetate gradient in heptane to give compound 16-4 as a colorless oil (375 mg, 88% yield).

[0137] Next, 10% Pd / C (38 mg) wetted with a minimum amount of water was added to a solution of Ph-Ala-OBn (compound 16-4; 375 mg, 1.32 mmol) in methanol (30 mL). The mixture was stirred under a hydrogen atmosphere (balloon) for 4 hours. The mixture was then filtered through a Celite pad and washed with methanol. The filtrate was concentrated under reduced pressure to obtain compound 17-4 as a colorless glass (glass; 254 mg, 99% yield), which was reacted with compound 14 according to the following reaction scheme 15.

[0138] [Reaction Scheme 15] [ka]

[0139] More specifically, HATU (106 mg, 0.278 mmol, 1.1 equiv) was added to a suspension of GlnThr(OBn)GlyThr(OBn)GlyLys(Cbz)Thr(OBn)OBn (compound 14; 300 mg, 0.253 mmol, 1 equiv) and Ph-Ala-OH (compound 17-4; 49.0 mg, 0.253 mmol, 1) in N,N-diisopropylethylamine (97.0 μL, 0.556 mmol, 2.2 equiv) and dichloromethane (20 mL). The mixture was stirred at room temperature for 2 h, and the reaction mixture was washed with NaHCO3 (saturated aqueous). The organic matter was concentrated under reduced pressure, and the obtained residue was purified with a gradient of 40 to 100% acetonitrile (0.1% formic acid) in water (0.1% formic acid) (60 g) on ​​a C18 column. After lyophilization, compound 18-4 was obtained as a colorless solid (200 mg, 58%).

[0140] Then, 10% Pd / C (110 mg) was added to a solution of Ph-AlaGlnThr(OBn)GlyThr(OBn)GlyLys(Cbz)Thr(OBn)OBn (compound 18-4; 110 mg, 80.8 μmol) in 2 M hydrochloric acid (aqueous, 1.0 mL) and 2-propanol (20 mL). The mixture was stirred under a hydrogen atmosphere (balloon) for 18 hours. The mixture was filtered through a 0.45 μm syringe filter. The filtrate was concentrated under reduced pressure, and the residue was dissolved in water and then lyophilized. The dried material was purified on a 60 g C18 column using a 5-50% acetonitrile (0.1% formic acid) gradient in water (0.1% formic acid) and lyophilized to obtain compound 19-4 as a colorless solid (23.2 mg, 33% yield).

[0141] Compound 19-4 (Ph-AQTGTGKT) 1 H NMR data was measured as follows: 1H NMR (400MHz; D2O): δ=7.74-7.70(m, 2H), 7.58-7.53(m, 1H), 7.48-7.43(m , 2H), 4.45-4.34(m, 3H), 4.32-4.27(m, 2H), 4.25-4.14(m, 3H), 4.11(d, J 3.8Hz, 1H), 4.00-3.96(m, 2H), 3.89(s, 2H), 2.91(t, J 7.4Hz, 2H), 2.34(t, J 7.6Hz, 2H), 2.14-2.06(m, 1H), 2.01-1.91(m, 1H), 1.85-1.76(m, 1H), 1.71-1.56(m, 3H), 1.43(d, J 7.3Hz, 3H), 1.40-1.30(m, 2H), 1.16-1.07(m, 9H), 16 exchangeable protons not visible.

[0142] 1.7.5. Synthesis of Naphthyl-AQTGTGKT Naphthyl-AQTGTGKT (compound 19-5) was synthesized by reacting compound 17-5 obtained according to the following reaction formula 16 with compound 14 according to reaction formula 17 to synthesize the final target compound, Naphthyl-AQTGTGKT.

[0143] [Reaction Scheme 16] [ka]

[0144] More specifically, H-Ala-OBzl.HCl (388 mg, 1.80 mmol, 1.2 equiv.) was suspended in ethyl acetate (10 mL) and N,N-diisopropylethylamine (653 μL, 3.75 mmol, 2.5 equiv.) was added. After stirring at room temperature for 5 min, HATU (855 mg, 2.25 mmol, 1.5 equiv.) and 2-naphthoic acid (258 mg, 1.50 mmol, 1 equiv.) were added and the mixture was stirred at room temperature for 2 h. The reaction mixture was diluted with ethyl acetate and washed with NH4Cl (saturated aqueous), NaHCO3 (saturated aqueous), and brine. The organics were dried (Na2SO4), filtered, and concentrated under reduced pressure. The residue was purified on a 25 g column with a 2-40% ethyl acetate gradient in heptane to give compound 16-5 as a colorless solid (385 mg, 77% yield).

[0145] Then, 10% Pd / C (39 mg) wetted with a minimum amount of water was added to a solution of 2-Naphthyl-AlaOBn (compound 16-5; 385 mg, 1.15 mmol) in methanol (20 mL). The mixture was stirred under a hydrogen atmosphere (balloon) for 4 hours. The mixture was then filtered through a Celite pad and washed with methanol. The filtrate was concentrated under reduced pressure to obtain compound 17-5 as a colorless solid (266 mg, 95% yield), which was reacted with compound 14 according to the following reaction scheme 17.

[0146] [Reaction Scheme 17] [ka]

[0147] More specifically, HATU (106 mg, 0.278 mmol, 1.1 equiv) was added to a suspension of GlnThr(OBn)GlyThr(OBn)GlyLys(Cbz)Thr(OBn)OBn (compound 14; 300 mg, 0.253 mmol, 1 equiv) and 2-Naphthyl-Ala-OH (compound 17-5; 61.0 mg, 0.253 mmol, 1 equiv) in N,N-diisopropylethylamine (97.0 μL, 0.556 mmol, 2.2 equiv) and dichloromethane (20 mL). The mixture was stirred at room temperature for 2 h and then washed with (saturated aqueous) NaHCO3. The organic layer obtained was concentrated under reduced pressure, and the residue was purified using a 60 g gradient of 40-100% acetonitrile (0.1% formic acid) in water (0.1% formic acid) on a C18 column. After lyophilization, compound 18-5 was obtained as a colorless solid (230 mg, 64% yield).

[0148] Then, 10% Pd / C (101 mg) was added to a solution of 2-Naphthyl-AlaGlnThr(OBn)GlyThr(OBn)GlyLys(Cbz)Thr(OBn)OBn (compound 18-5; 101 mg, 71.5 μmol) in 2 M hydrochloric acid (aqueous, 1.0 mL) and 2-propanol (20 mL). The mixture was stirred under hydrogen atmosphere (balloon) for 3 h. The mixture was filtered through a 0.45 μm syringe filter. The obtained filtrate was concentrated under reduced pressure, and the residue was dissolved in water and then lyophilized. The dried material was purified on a C18 column with a 5-40% acetonitrile (0.1% formic acid) gradient in water (0.1% formic acid) for 30 g, and lyophilized to obtain compound 19-5 as a colorless solid (23.2 mg, 33% yield).

[0149] Compound 19-5 (Naphthyl-AQTGTGKT) 1 H NMR data was measured as follows: 1H NMR (400MHz; D2O): δ=8.32(s, 1H), 8.01-7.90(m, 3H), 7.79-7.73(m, 1H) , 7.64-7.55(m, 2H), 4.51-3.70(m, 13H), 2.96-2.81(m, 2H), 2.39-2.30( m, 2H), 2.18-2.04(m, 1H), 2.03-1.93(m, 1H), 1.85-1.72(m, 1H), 1.71-1 .53(m, 3H), 1.50-1.43(m, 3H), 1.39-1.24(m, 2H), 1.19-1.04(m, 9H), 16 exchangeable protons not visible.

[0150] 1.8.Synthesis of Ac-AQTGTGKT Ac-AQTGTGKT (compound 19-6) was synthesized according to the following reaction scheme 18 to synthesize the final target compound, Ac-AQTGTGKT.

[0151] [Reaction Scheme 18] [ka]

[0152] More specifically, HATU (112 mg, 0.294 mmol, 1.2 equiv) was added to a suspension of GlnThr(OBn)GlyThr(OBn)GlyLys(Cbz)Thr(OBn)OBn (compound 14; 300 mg, 0.245 mmol, 1 equiv) and Ac-Ala-OH (32.1 mg, 0.245 mmol, 1 equiv) in N,N-diisopropylethylamine (94.0 μL, 0.540 mmol, 2.2 equiv) and dichloromethane (30 mL). The mixture was stirred at room temperature for 14 h, and the reaction mixture was washed with (saturated aqueous) NH4Cl, NaHCO3, and water. The organic layer was concentrated under reduced pressure, and the residue was purified on a 60 g, C18 column with a 50-95% acetonitrile (0.1% formic acid) gradient in water (0.1% formic acid). The desired fractions were combined and lyophilized to afford compound 18-6 as a colorless solid (104 mg, 33% yield).

[0153] Then, 10% Pd / C (10 mg) was added to a solution of Ac-AlaGlnThr(OBn)GlyThr(OBn)GlyLys(Cbz)Thr(OBn)OBn (compound 18-6; 33 mg, 25 μmol) in 2 M hydrochloric acid (aqueous, 0.19 mL) and 2-propanol (5 mL). The mixture was stirred under a hydrogen atmosphere (balloon) for 14 h. The mixture was filtered through a 0.45 μm syringe filter. The filtrate was concentrated under reduced pressure, dissolved in water, and lyophilized. The dried material was purified on a 500 mg SCX-2 cartridge, eluting with 0.5 M ammonia in methanol. The desired fractions were combined, concentrated under reduced pressure, and lyophilized to give compound 19-6 as a colorless solid (7.6 mg, 37% yield).

[0154] Compound 19-6 (Ac-AQTGTGKT) 1 H NMR data was measured as follows: 1 H NMR (400MHz; D2O): δ=4.40-4.33(m, 2H), 4.32-4.27(m, 2H), 4.24-4.12(m, 4H), 4.08(d, J 4.0Hz, 1H), 4.03-3.88(m, 4H), 2.92(t, J 7.2Hz, 2H), 2.32(t, J 7.8Hz, 2H), 2.15-2.02(m, 1H), 1.99-1.88(m, 4H), 1.86-1.76(m, 1H), 1.74-1.55(m, 3H), 1.45-1.31(m, 2H), 1.29(t, J 7.4Hz, 2H), 1.20-1.06(m, 10H), 16 exchangeable protons not visible.

[0155] 1.9.4-Synthesis of PhPh-AQTGTGKT 1.9.1. Synthesis of QTGTGKT intermediate The intermediate QTGTGKT for the synthesis of 4-PhPh-AQTGTGKT was synthesized according to the following reaction schemes 19 to 23:

[0156] [Reaction Scheme 19] [ka]

[0157] [Reaction Scheme 20] [ka]

[0158] [Reaction Scheme 21] [ka]

[0159] [Reaction Scheme 22] [ka]

[0160] [Reaction Scheme 23] [ka]

[0161] The above-mentioned Reaction Schemes 19 to 23 are almost similar to the above-mentioned Reaction Schemes 1 to 5, except for the presence or absence of a benzyl protecting group, and therefore a duplicated explanation will be omitted.

[0162] Synthesis of 1.9.2.4-PhPh-AQTGTGKT The compound 31 obtained in the above Reaction Scheme 23 was combined with the alanine derivative synthesized in the following Reaction Scheme 24 according to Reaction Scheme 25 to obtain the final product 4-PhPh-AQTGTGKT.

[0163] [Reaction Scheme 24] [ka]

[0164] [Reaction Scheme 25] [ka]

[0165] In addition, the above-mentioned reaction schemes 24 and 25 were carried out in a similar manner to the above-mentioned reaction schemes, and therefore the overlapping explanations will be omitted. Finally, a colorless solid (583 mg, 68% yield) of compound 36 was obtained.

[0166] Compound 36 (4-PhPh-AQTGTGKT) 1 H NMR data was measured as follows: 1 H NMR (400MHz; D2O): δ=7.85(d, J 8.8Hz, 2H), 7.77(d, J 8.8Hz, 2H), 7.70(d, J 7.2Hz, 2H), 7.49(t, J 7.2Hz, 2H), 7.42(t, J 7.1Hz, 1H), 4.34-4.04(m, 6H), 4.07(d, J 3.6Hz, 1H), 3.93(d, J 2.0Hz, 2H), 2.82(t, J 7.0Hz, 2H), 1.82-1.67(m, 1H), 1.66-1.55(m, 1H), 1.54-1.44(m, 2H), 1.37-1.22(m, 2H), 1.18(d, J 6.4Hz, 3H), 1.06(t, J 6.5Hz, 3H), 10 exchangeable protons not visible.

[0167] 1.10. Compound Verification The seven AQTGTGKT analogs obtained by the above-mentioned method are: 1 The chemical structures were confirmed by H NMR and UPLC-MS (ultraperformance liquid chromatography-mass spectrometry) techniques. The analytical results are shown in Figures 1 to 7, and the compounds are summarized in Table 1.

[0168] [Table 1] JPEG0007610862000049.jpg95160

[0169] Example 2: Confirmation of anticancer effect of AQTGTGKT analogue 2.1. Inhibitory effect on lung cancer cell activity Using the CTG analysis method described in the experimental method, the lung cancer cell line H1299 was treated with the AQTGTGKT analog at a concentration of 100 μM for 48 hours, and the lung cancer cell lines H820 and H1975 were treated with the AQTGTGKT analog at a concentration of 10 μM for 72 hours, and the cytotoxicity of each analog was compared.

[0170] As a result, as shown in Figure 8, when H1299 cell line was treated with 4-PhPh-AQTGTGKT, cell activity was reduced by approximately 10% compared to the control group (AQTGTGKT), while Ac-AQTGTGKT showed a 4.2% inhibitory effect on cell activity, 3-PhPh-AQTGTGKT showed a 15% inhibitory effect, 4-MeOPh-AQTGTGKT showed a 12% inhibitory effect, 2-PhPh-AQTGTGKT showed a 20% inhibitory effect, Ph-AQTGTGKT showed a 18% inhibitory effect, and Naphthyl-AQTGTGKT showed a 14% inhibitory effect on cell activity.

[0171] In the case of the H1975 cell line, as shown in Figure 9, when Ac-AQTGTGKT was treated, cell activity was reduced by approximately 16% compared to the control group (AQTGTGKT), while 3-PhPh-AQTGTGKT, 4-MeOPh-AQTGTGKT, and 2-PhPh-AQTGTGKT showed a 33%, 4% and 3% inhibitory effect on cell activity.

[0172] Furthermore, in the case of the H820 cell line, as shown in Figure 10, when treated with 3-PhPh-AQTGTGKT, cell activity was reduced by approximately 12% compared to the control group (AQTGTGKT), and 4-MeOPh-AQTGTGKT showed a 10% inhibitory effect on cell activity.

[0173] These results indicate that the amidated analogues can highly effectively inhibit the proliferation of lung cancer cells.

[0174] 2.2. Inhibitory effect on breast cancer cell activity and proliferation (1) Inhibitory effect on breast cancer cell activity Using the CTG analysis method described in the experimental method, breast cancer cell line MDA-MB-231 was treated with AQTGTGKT analog at a concentration of 100 μM for 48 hours, and then the intracellular ATP level was measured to compare the activity of the cells.

[0175] As a result, as shown in Figure 11a, when 4-PhPh-AQTGTGKT was treated, cell activity was reduced by about 30% compared to the control group (AQTGTGKT), demonstrating its excellent effect in inhibiting the division of breast cancer cells.

[0176] (2) Inhibitory effect on the growth and proliferation of breast cancer cells The breast cancer cell line was treated with AQTGTGKT analogs by the MTT assay described in the above experimental method, and the cell proliferation inhibitory effects were compared. HCC1937 was used as the breast cancer cell line.

[0177] HCC1937 cell line was treated with Ac-AQTGTGKT, 3-PhPh-AQTGTGKT, 4-MeOPh-AQTGTGKT, 2-PhPh-AQTGTGKT, Ph-AQTGTGKT, and Naphtyl-AQTGTGKT at a concentration of 100 μM, and the cell proliferation inhibitory effects were compared after 72 hours. As a result, as shown in Figure 11b, Ac-AQTGTGKT, 3-PhPh-AQTGTGKT, 4-MeOPh-AQTGTGKT, 2-PhPh-AQTGTGKT, Ph-AQTGTGKT, and Naphtyl-AQTGTGKT inhibited cell proliferation by approximately 11.1%, 17.1%, 14.3%, 15.9%, 13.6%, and 9.5%, respectively, compared to the control group (AQTGTGKT).

[0178] The above experimental results demonstrate that the AQTGTGKT analog according to the present invention exhibits excellent anti-cancer effects against breast cancer.

[0179] 2.3.Inhibitory effect on the growth and proliferation of blood cancer cells The cytotoxicity of the AQTGTGKT analog was compared by the MTT assay described in the Experimental Methods. The Jurkat clone E6-1 was used as the blood cancer cell line.

[0180] Jurkat clone E6-1 was treated with 4-PhPh-AQTGTGKT, Ac-AQTGTGKT, 3-PhPh-AQTGTGKT, 4-MeOPh-AQTGTGKT, 2-PhPh-AQTGTGKT, Ph-AQTGTGKT, and Naphtyl-AQTGTGKT at a concentration of 10 μM each, and the cell growth inhibitory effects were compared after 48 hours. As a result, as shown in Figure 12, it was shown that 4-PhPh-AQTGTGKT, Ac-AQTGTGKT, 3-PhPh-AQTGTGKT, 4-MeOPh-AQTGTGKT, 2-PhPh-AQTGTGKT, Ph-AQTGTGKT, and Naphtyl-AQTGTGKT inhibited cell proliferation by approximately 4.8%, 7.9%, 6.4%, 4%, 6.7%, 10.2%, and 10.4%, respectively, compared to the control group (AQTGTGKT).

[0181] The above results demonstrate that the AQTGTGKT analog according to the present invention has excellent anti-cancer effects against blood cancer cells.

[0182] 2.4. Inhibitory effect on the growth and proliferation of pancreatic cancer cells According to the MTT assay described in the above experimental method, the pancreatic cancer cell line CFPAC-1 was treated with the AQTGTGKT analog at a concentration of 10 μM for 72 hours, and the cytotoxicity was compared.

[0183] As a result, as shown in Figure 13, when 4-PhPh-AQTGTGKT was treated, cell activity decreased by about 12% compared to the control group (AQTGTGKT), while 3-PhPh-AQTGTGKT, 4-MeOPh-AQTGTGKT, Ph-AQTGTGKT, and Naphthyl-AQTGTGKT showed a cell activity inhibitory effect of 2%, 6%, 3%, and 3%, respectively.

[0184] The above results indicate that the AQTGTGKT analog according to the present invention has excellent anti-cancer effects against pancreatic cancer cells.

[0185] 2.5. Inhibitory effect on growth and proliferation of colon cancer cells According to the MTT assay described in the above experimental method, colon cancer cell line HT29 was treated with AQTGTGKT analogs at a concentration of 50 μM for 72 hours to compare cytotoxicity.

[0186] As a result, as shown in Figure 14, when 4-PhPh-AQTGTGKT was treated, cell proliferation was reduced by about 8% compared to the control group (AQTGTGKT), while Ac-AQTGTGKT showed a 58% cell proliferation inhibitory effect, 3-PhPh-AQTGTGKT showed a 59% cell proliferation inhibitory effect, 48% cell proliferation inhibitory effect, 2-PhPh-AQTGTGKT showed a 7% cell proliferation inhibitory effect, Ph-AQTGTGKT showed a 40% cell proliferation inhibitory effect, and Naphthyl-AQTGTGKT showed a 58% cell proliferation inhibitory effect. The above results indicate that the AQTGTGKT analog according to the present invention has an excellent anticancer effect against colon cancer cells.

[0187] 2.6. Tumor growth suppression effect in animal models inoculated with lung cancer cells According to the animal experimental analysis method described in the experimental method, nude mice implanted with the lung cancer cell line H820 were treated with the AQTGTGKT analog at a dose of 10 mpk five times at 3-day intervals and five times at 2-day intervals, for a total of 10 times, and the tumor growth was compared.

[0188] As a result, as shown in Figure 15, when the H820 lung cancer cell line was transplanted and treated with 3-PhPh-AQTGTGKT, tumor growth was reduced by approximately 16.5% compared to the control group (AQTGTGKT), while 2-PhPh-AQTGTGKT, Ph-AQTGTGKT, and Naphthyl-AQTGTGKT showed a tumor growth inhibitory effect of 27%, 20%, and 26%, respectively.

[0189] In addition, nude mice implanted with H1975 lung cancer cells were treated with the AQTGTGKT analog at a dose of 10 mpk for 14 days and tumor growth was compared.

[0190] As a result, as shown in FIG. 16, when 3-PhPh-AQTGTGKT was treated, the tumor growth was reduced by about 16.5% compared to the control group (AQTGTGKT), indicating an inhibitory effect on tumor growth.

[0191] Taken together, the above results demonstrate that the compounds of the present invention have excellent anti-cancer effects in an animal model implanted with lung cancer cells.

[0192] 2.7. Tumor growth suppression effect in animal models inoculated with breast cancer cells Using the animal experimental analysis method described in the experimental method, AQTGTGKT and three AQTGTGKT analogs (3-PhPh-AQTGTGKT, 4-MeOPh-AQTGTGKT, and Ph-AQTGTGKT) were administered at a dose of 10 mpk via the tail vein at two-day intervals for a total of seven times to immunodeficient mice implanted with breast cancer cell lines, and tumor growth was compared.

[0193] On the 15th day, two days after the seventh administration of the analog, the tumor volume was calculated to be 1884.17 mm on average in the AQTGTGKT group. 3 , 3-PhPh-AQTGTGKT group 944.70 mm 3 , 4-MeOPh-AQTGTGKT group: 812.69 mm 3 , Ph-AQTGTGKT group was 1133.80 mm 3 The tumor volumes in the AQTGTGKT group were approximately 49.86%, 56.86%, and 39.82%, respectively, and tumor growth was suppressed (FIG. 17).

[0194] Taken together, the above results indicate that the compounds of the present invention have excellent anti-cancer effects in an animal model implanted with breast cancer cells, without affecting the body weight of the mice.

[0195] 2.8. Tumor growth suppression effect in an animal model inoculated with colon cancer cells Mice implanted with CT26 cell lines in which CAGE gene expression had been induced using the animal experimental analysis method described in the experimental method were administered AQTGTGKT and two AQTGTGKT analogs (3-PhPh-AQTGTGKT and Naphthyl-AQTGTGKT) at a dose of 10 mpk each, a total of seven times at two-day intervals, via the tail vein, and tumor growth was compared.

[0196] As a result, it was shown that tumor growth was suppressed in the analog administration group from 10 days after administration compared to the control group (AQTGTGKT administration group). The tumor volume was calculated on the 14th day, the day after the seventh administration of the analog, and the average tumor volume in the AQTGTGKT administration group was 2573.07 mm 3 The 3-PhPh-AQTGTGKT group had a mean of 880.35 mm 3 The Naphthyl-AQTGTGKT group had an average of 880.35 mm 3 It was confirmed that the tumor growth was suppressed by approximately 65.79% in the 3-PhPh-AQTGTGKT group and approximately 62.04% in the Naphthyl-AQTGTGKT group compared to the AQTGTGKT group (Figure 18). At the 12th and 14th day baseline, tumor growth was suppressed statistically significantly (p<0.001) in the 3-PhPh-AQTGTGKT and Naphthyl-AQTGTGKT analogue groups compared to the AQTGTGKT group.

[0197] Overall, the above results indicate that the compounds of the present invention have excellent anti-cancer effects in an animal model transplanted with CT26 cells induced to express CAGE, without affecting the body weight of the mice.

[0198] Example 3: Confirmation of stability of AQTGTGKT analog in blood In order to confirm how long the AQTGTGKT analog of the present invention remains stable in blood, 10 μM of each compound was added to 100% human blood, and the amount remaining in the blood was measured at 0, 5, 10, 15, 30, 60 and 120 minutes.

[0199] As a result, as shown in FIG. 19 to FIG. 26, in the case of AQTGTGKT, the half-life was less than 1 minute, and it was confirmed that it was almost decomposed in the blood within 20 minutes, and the residual AQTGTGKT was hardly detected. In contrast, all of the seven AQTGTGKT analogs according to the present invention showed significantly improved stability, and in particular, 4-PhPh-AQTGTGKT, 3-PhPh-AQTGTGKT, 4-MeOPh-AQTGTGKT, 2-PhPh-AQTGTGKT, Ph-AQTGTGKT and Naphthyl-AQTGTGKT showed a half-life of 47 to 105 minutes, and even after 120 minutes, 20 to 40% remained in the blood, showing high stability. These results indicate that the amidated analogs not only show a better anticancer effect, but also show a further improved stability in the blood, and can exist stably without being decomposed until they reach the target.

[0200] The above description of the present invention is for illustrative purposes only, and those skilled in the art will understand that the present invention can be easily modified into other specific forms without changing the technical concept or essential features of the present invention. Therefore, the above-described embodiments should be understood to be illustrative in all respects and not restrictive. [Industrial Applicability]

[0201] The present invention relates to a novel oligopeptide AQTGTGKT analog compound, a pharmaceutical composition for preventing or treating cancer containing the same as an active ingredient, and a method for preparing the same, and it has been confirmed that the oligopeptide AQTGTGKT analog exhibits excellent anti-cancer effects and is stable in human blood. Therefore, the pharmaceutical composition according to the present invention has the advantages of oligopeptides, that is, a smaller molecular weight than antibodies, less risk of immune reaction, and easy penetration into tissues, and is also excellent in the effect of inhibiting the proliferation of cancer cells and exhibits the effect of being stable in human blood, so it is expected to be used as a useful anti-cancer agent for treating cancer.

Claims

1. A compound represented by the following general formula: [General formula] X-AQTGTGKT (In the above general formula, A is alanine, Q is glutamine, T is threonine, G is glycine, and K is lysine. X is, 【Chemistry 1】 and is bound to the N-terminus of A.

2. A pharmaceutical composition for preventing or treating cancer, comprising as an active ingredient a compound represented by the following general formula: [General formula] X-AQTGTGKT (In the above general formula, A is alanine, Q is glutamine, T is threonine, G is glycine, and K is lysine. X is, 【Chemistry 2】 and is bound to the N-terminus of A.

3. The pharmaceutical composition of claim 2, wherein the cancer is selected from the group consisting of lung cancer, breast cancer, blood cancer, colon cancer, pancreatic cancer, and combinations thereof.

4. The pharmaceutical composition according to claim 3, wherein the lung cancer is non-small cell lung cancer.

5. 4. The pharmaceutical composition of claim 3, wherein the blood cancer is a blood cancer selected from the group consisting of leukemia, lymphoma, multiple myeloma, and combinations thereof.

6. The X is 【Chemistry 3】 The pharmaceutical composition according to claim 2, characterized in that the cancer is lung cancer.

7. The X is 【Chemistry 4】 and the cancer is breast cancer.

8. The X is 【Chemistry 5】 and the cancer is a blood cancer.

9. The X is 【Chemistry 6】 3. The pharmaceutical composition according to claim 2, wherein the cancer is pancreatic cancer.

10. The X is 【Chemistry 7】 3. The pharmaceutical composition according to claim 2, wherein the cancer is colorectal cancer.

11. The X is 【Chemistry 8】 The pharmaceutical composition according to claim 2, characterized in that, when the compound is, the half-life of the compound in human blood is 100 to 150 minutes.

12. The X is 【Chemistry 9】 The pharmaceutical composition according to claim 2, characterized in that when the compound is one selected from the group consisting of:

13. The X is 【Chemistry 10】 The pharmaceutical composition according to claim 2, characterized in that, when the compound is, the half-life of the compound in human blood is 20 to 30 minutes.

14. A method for preparing oligopeptide X-AQTGTGKT comprising the steps of: (A is alanine, Q is glutamine, T is threonine, G is glycine, and K is lysine, The X is 【Chemistry 11】 It is one selected from the group consisting of. (1) synthesizing TG and KT, respectively; (2) synthesizing TGKT by binding TG and KT; (3) synthesizing TGTGKT by binding TG to the N-terminus of the TGKT; (4) synthesizing QTGTGKT by binding Q to the N-terminus of the TGTGKT; and (5) synthesizing X-AQTGTGKT by binding an alanine derivative (XA) to the N-terminus of QTGTGKT.

15. 13. A method for preventing or treating cancer, comprising administering a compound of claim 1 to an individual in need thereof, wherein the subject is a non-human subject.

16. 13. Use of a compound according to claim 1 for the manufacture of a medicament for the treatment of cancer.

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