WO2025146620A1 - Anti-cancer combination comprising anti-c-met antibody - Google Patents

Abstract

The present invention relates to: a combination comprising an antibody, or an antigen-binding fragment thereof, that specifically binds to a hepatocyte growth factor receptor (c-Met), and a second anticancer agent; and a use of the combination for the prevention and treatment of cancer. The combination therapy of the anti-c-Met antibody and the second anticancer agent of the present invention has excellent anticancer activity and exhibits a synergistic effect that can kill cancer cells even in cancers that are difficult to overcome due to drug resistance, and thus can be very usefully employed for the prevention and treatment of cancer.

Description

Description of the Invention

【Title of invention】Anti-cancer combination comprising anti-c-Met antibody

【Technical Field】 The present invention relates to a combination comprising an antibody or an antigen-binding fragment thereof that specifically binds to hepatocyte growth factor receptor (c-Met) and a second anticancer agent, and a use of the combination for preventing or treating cancer.

【Background Art】 Gastric cancer is one of the most common cancers, ranking third in the world in terms of cancer incidence (Int. J. Cancer, 2015, 136: E359). Several anticancer drugs, such as cisplatin, have been developed and are being used in the treatment of various cancer types or are considered as anticancer candidates. However, unexpected side effects, such as anticancer drug resistance, continue to be reported (Cirt. Rev. Oncol. Hematol., 2002, 42: 317). The present inventors have strived to develop an anticancer treatment with excellent anticancer activity and reduced side effects, and have completed the present invention by confirming the excellent efficacy of combination therapy using a c-Met antibody.

【National Research and Development Project Supporting This Invention】 【Project ID】 1711200539

【Assignment Number】DD118094

Ministry of Science and ICT, Ministry of Trade, Industry and Energy, Ministry of Health and Welfare

【Name of Project Management (Professional) Institution】 National Research Foundation of Korea, Korea Institute of Industrial Technology Evaluation and Planning, Korea Health Industry Development Institute

【Research Project Name】National New Drug Development Project

【Research Project Name】Non-small cell lung cancer targeting dual antibody new drug

【Name of the project performing organization】 Chong Kun Dang Co., Ltd.

Research Period: September 1, 2021 - December 31, 2023

【Contents of the invention】

【Technical Challenge】 The present inventors have completed the present invention by confirming that when an anti-c-Met antibody and a second anticancer drug are administered together, a synergistic effect is achieved, and significantly superior anticancer activity is exhibited even against cancer cells that are difficult to treat with existing drugs.

【Technical Solution】 One object of the present invention is to provide a pharmaceutical composition for preventing or treating cancer, comprising an antibody or an antigen-binding fragment thereof that specifically binds to hepatocyte growth factor receptor (c-Met; hepatocyte growth factor receptor), and a second anticancer agent as active ingredients. Another object of the present invention is to provide a pharmaceutical composition for preventing or treating cancer, comprising as an active ingredient an antibody or an antigen-binding fragment thereof that specifically binds to c-Met, wherein the pharmaceutical composition is administered in combination with a second anticancer agent to a subject. Another object of the present invention is to provide a method for preventing or treating cancer, comprising a step of administering to a subject in need of prevention or treatment of cancer an antibody or an antigen-binding fragment thereof that specifically binds to c-Met, wherein the antibody or the antigen-binding fragment thereof is administered in combination with a second anticancer agent. Another object of the present invention is to provide a use of an antibody or an antigen-binding fragment thereof that specifically binds to c-Met for preventing or treating cancer, wherein the antibody or the antigen-binding fragment thereof is administered in combination with a second anticancer agent. Another object of the present invention is to provide a use of an antibody or antigen-binding fragment thereof that specifically binds to c-Met in the manufacture of a medicament for use in preventing or treating cancer, wherein the antibody or antibody fragment is administered in combination with a second anticancer agent. Specific embodiments of the present invention are as follows, but the scope of the present invention is not limited to these embodiments.

[1] The pharmaceutical composition for preventing or treating cancer according to the present invention comprises (A) a compound selected from the group consisting of SEQ ID NO: 313, SEQ ID NO: 314, SEQ ID NO: 315 and SEQ ID NO: 316. An antibody or antigen-binding fragment thereof that specifically binds to hepatocyte growth factor receptor (c-Met) that binds to one or more epitope regions described by a selected amino acid sequence, and (B) comprises at least one of the following (1) drug 1 and (2) drug 2 as an active ingredient:

(1) Drug 1: SN-38 represented by the following chemical formula 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof

[Chemical Formula 1]

(2) Drug 2: Irinotecan represented by the following chemical formula 2, an optical isomer thereof, or a pharmaceutically acceptable salt thereof

[2] 상기 [1]에 있어서 , 상기 (B)는 상기 약물 1일 수 있다. [Chemical Formula 2]

[2] In the above [1], the (B) may be the drug 1.

[3] In the above [1], the (B) may be the drug 2.

[4] In any one of the above [1] to [3], the cancer may be at least one of stomach cancer, colon cancer, and esophageal cancer.

[5] In any one of the above [1] to [4], the cancer may be gastric cancer.

[6] In any one of [1] to [5] above, the cancer may be primary, recurrent, progressive or metastatic cancer.

[7] In any one of the above [1] to [6], the cancer may be a cancer resistant to chemotherapy.

[8] In the above, the chemotherapy may be fluorouracil, capecitabine, oxaliplatin, epirubicin, cisplatin, carboplatin, etoposide, doxorubicin, methotrexate, leucovorin, irinotecan, docetaxel, paclitaxel, trifluridine/tipiracil, or a combination thereof.

[9] In any one of the above [1] to [8], the cancer may be a c-Met wild-type cancer or a cancer that overexpresses c-Met, amplifies c-Met, or activates c-Met.

[1] In any one of the above [1] to [9], the c-Met is represented by the sequence number It may be indicated by the amino acid sequence described as 317.

[11] In any one of the above [1] to [1], the antibody may bind to an epitope region described by the amino acid sequence of SEQ ID NO: 313, SEQ ID NO: 314, SEQ ID NO: 315 and SEQ ID NO: 316.

[12] In any one of the above [1] to [11], the antibody or antigen-binding fragment thereof binds to human c-Met with a KD of 1 x 10' ⁷ M or less, wherein the KD may be determined by surface plasmon resonance (Biacore) analysis.

[13] In any one of the above [1] to [12], the antibody may be an antibody comprising a light chain variable region including a light chain CDR1 as set forth in SEQ ID NO: 1; a light chain CDR2 as set forth in SEQ ID NO: 2; a light chain CDR3 as set forth in SEQ ID NO: 3, and a heavy chain variable region including a heavy chain CDR1 as set forth in SEQ ID NO: 4; a heavy chain CDR2 as set forth in SEQ ID NO: 5; and a heavy chain CDR3 as set forth in SEQ ID NO: 6, or an affinity-optimized antibody thereof.

[14] In any one of the above [1] to [13], the antibody may include a light chain variable region described in SEQ ID NO: 7 and a heavy chain variable region described in SEQ ID NO: 8.

[15] In any one of the above [1] to [14], the antibody may comprise (a) a light chain variable region described in SEQ ID NO: 11 and a heavy chain variable region described in SEQ ID NO: 13; or (b) a light chain variable region described in SEQ ID NO: 12 and a heavy chain variable region described in SEQ ID NO: 14. [16] In any one of the above [1] to [15], the antibody may include a hinge region described in any one of SEQ ID NO: 19 to SEQ ID NO: 26.

[17] In any one of the above [1] to [16], the antibody comprises a light chain variable region comprising a light chain CDR1 as set forth in SEQ ID NO: 1; a light chain CDR2 as set forth in SEQ ID NO: 2; a light chain CDR3 as set forth in SEQ ID NO: 3, and a heavy chain variable region comprising a heavy chain CDR1 as set forth in SEQ ID NO: 4; a heavy chain CDR2 as set forth in SEQ ID NO: 5; and a heavy chain CDR3 as set forth in SEQ ID NO: 6, wherein at least one amino acid sequence is substituted in the antibody, wherein (i) G at the 1st position of the light chain CDR1 is A, E, K, L, N, R, S, V, or W; A at the 2nd position is C, G, I, P, S, T, or V; S at the 3rd position is G, M, N, P, Q, R, S, or T; E at the 4th position is A, D, F, G, H, K, M, Q, R, S, T, or V; N at the 5th position is A, D, E, G, K, L, P, Q, R, S, T or V; I in the 6th position is A, F, L, M, Q, R, S, T or V; Y in the 7th position is F, H, R or V; or G in the 8th position is D, F, H, M, N, R, S, T or

Substitution with V; (ii) G at position 1 of light chain CDR2 is D, F, H, K, P, Q, S, V or Y;

T at the 3rd position is replaced with Q; or N at the 4th position is replaced with ; (iii) Q at the 1st position of the light chain CDR3 is replaced with E, G, I, M or N; 凡 at the 2nd position is replaced with A, D, E, H, L, Q, S or T; V at the 3rd position is replaced with I, L, M, N, Q, S or T; L at the 4th position is replaced with F, H, I, M, R, S, V, W or Y; S at the 5th position is replaced with C, D, E, F, G, H, K, L, N, Q, R, T, V or Y; S at the 6th position is replaced with D, E, F, G, H, I, L, M, N, P, Q, R, T, V or Y; P at the 7th position is replaced with

A, D, E, G, N, Q, S or V; Y in the 8th position is E, F, L, M or Q; or Y in the 9th position is T at position 1 is replaced with D, F, G, I, L, N, S, V, W or Y; (iv) 日 at position 1 of heavy chain CDR1 is replaced with G or Q; Y at position 2 is replaced with Q; or I at position 4 is replaced with A or Y; (v) The 3rd position of the heavy chain CDR2 is D, E, W, or Y; the 5th position is D, H, or Y; the 6th position is F, P, W, or Y; the G at the 7th position is A, F, L, N, or T; the 8th position is F, P, S, T, or Y; the T at the 9th position is A, D, E, F, G, H, L, P, S, or V; the H at the 10th position is A, D, F, M, R, S, T, V, W, or Y; the F at the 11th position is G, H, I, L, M, N, P, Q, V, or Y; the S at the 12th position is A, D, G, H, I, L, P, T, or V; the A at the 13th position is D, E, F, G, H, I, K, L, M, P, R, S, T, V, or Y; R in the 14th position is replaced by A, E, G, H, L, N, P, Q, S, W or Y; F in the 15th position is replaced by D, E, G, L, M, P, R, S, V or W; V in the 16th position is replaced by A, E, F, G, H, L, R, S, T, V or Y; or G in the 17th position is replaced by E, F, H, L, M, N, P, Q, R, S, T, V or W; Or (vi) the heavy chain CDR3 may include G at the 1st position replaced by E, F, H, N, Q, V or W; 日 at the 2nd position replaced by E; 오 at the 3rd position replaced by L, Q, T or V; 으 at the 4th position replaced by W; F at the 5th position replaced by L or Y; L at the 6th position replaced by Q, S or Y; or Y at the 7th position replaced by C, L, M, N or Y, wherein the light chain CDR1 may include 0 to 5 substitutions, the light chain CDR2 may include 0 to 1 substitution, the light chain CDR3 may include 0 to 7 substitutions, the heavy chain CDR1 may include 0 to 1 substitution, the heavy chain CDR2 may include 0 to 11 substitutions, and the heavy chain CDR3 may include 0 to 6 substitutions.

[18] In any one of the above [13] to [17], the affinity-optimized antibody comprises a light chain CDR1 as described in SEQ ID NO: 1 and any one of SEQ ID NOs: 211 to 250; any one of SEQ ID NO: 2, SEQ ID NOs: 164 to 172, SEQ ID NO: 209 and SEQ ID NO: 210. A light chain variable region comprising a light chain CDR2 described as one; a light chain CDR3 described as one of SEQ ID NO: 3, SEQ ID NO: 124 to 163, SEQ ID NO: 173 to 208, and SEQ ID NO: 251 to 283 and a heavy chain CDR1 described as one of SEQ ID NO: 4 and SEQ ID NO: 90 to 94; a heavy chain CDR2 described as one of SEQ ID NO: 5, SEQ ID NO: 36 to 45, SEQ ID NO: 54 to 89, and SEQ ID NO: 100 to 123; and a heavy chain variable region comprising a heavy chain CDR3 described as one of SEQ ID NO: 6, SEQ ID NO: 46 to 53, and SEQ ID NO: 95 to 99.

[19] In any one of the above [13] to [18], the affinity-optimized antibody may comprise a light chain variable region described in any one of SEQ ID NOs: 11 and 288 to 293 and a heavy chain variable region described in any one of SEQ ID NOs: 13 and 284 to 287.

[2] In any one of the above [13] to [19], the affinity-optimized antibody comprises (a) a light chain variable region as set forth in SEQ ID NO: 11 and a heavy chain variable region as set forth in SEQ ID NO: 284; (b) a light chain variable region as set forth in SEQ ID NO: 11 and a heavy chain variable region as set forth in SEQ ID NO: 287; (c) a light chain variable region as set forth in SEQ ID NO: 292 and a heavy chain variable region as set forth in SEQ ID NO: 13; (d) a light chain variable region as set forth in SEQ ID NO: 290 and a heavy chain variable region as set forth in SEQ ID NO: 287; (e) a light chain variable region as set forth in SEQ ID NO: 288 and a heavy chain variable region as set forth in SEQ ID NO: 285; (f) a light chain variable region as set forth in SEQ ID NO: 289 and a heavy chain variable region as set forth in SEQ ID NO: 286; (g) a light chain variable region as set forth in SEQ ID NO: 290 and a heavy chain variable region as set forth in SEQ ID NO: 286; (h) a light chain variable region as set forth in SEQ ID NO: 291 A light chain variable region and a heavy chain variable region as set forth in SEQ ID NO: 286; (i) a light chain variable region as set forth in SEQ ID NO: 293 and a heavy chain variable region as set forth in SEQ ID NO: 286; or (j) a light chain variable region as set forth in SEQ ID NO: 288 and a heavy chain variable region as set forth in SEQ ID NO: 284.

[21] In any one of [1] to [2], the antibody may further comprise an antibody or an antigen-binding fragment thereof that specifically binds to epidermal growth factor receptor (EGFR).

[22] In any one of the above [1] to [21], the antibody may be one in which an antibody or an antigen-binding fragment thereof that binds to epidermal growth factor receptor (EGFR) is linked to one end of a light chain or heavy chain of an antibody that specifically binds to c-Met.

[23] In the above [21] or [22], the antigen-binding fragment binding to the EGFR may be Fab, Fab', F(ab')2 or Fv.

[24] In the above [23], the Fv may be one or more scFv fragments selected from the group consisting of Erbitux, Vectibix, Portrazza, and TheraCIM.

[25] In the above [24], the Erbitux scFv may include an amino acid sequence described as SEQ ID NO: 295 or SEQ ID NO: 296.

[26] In the above [24], the Vectibix scFv may include an amino acid sequence described as SEQ ID NO: 297.

[27] In any one of the above [21] to [26], the connection of the antibody or antigen-binding fragment thereof that binds to the EGFR is connected by a connector described in SEQ ID NO: 294. It could be.

[28] In any one of [1] to [27] above, the antibody or antigen-binding fragment thereof may include a light chain variable region as described in SEQ ID NO: 11 and any one of SEQ ID NOs: 288 to 293; and a heavy chain variable region as described in any one of SEQ ID NOs: 298 to 312.

[29] In any one of [1] to [28] above, the antibody or antigen-binding fragment thereof comprises (1) a light chain variable region as set forth in SEQ ID NO: 11 and a heavy chain variable region as set forth in SEQ ID NO: 298; (2) a light chain variable region as set forth in SEQ ID NO: 11 and a heavy chain variable region as set forth in SEQ ID NO: 299; (3) a light chain variable region as set forth in SEQ ID NO: 11 and a heavy chain variable region as set forth in SEQ ID NO: 302; (4) a light chain variable region as set forth in SEQ ID NO: 292 and a heavy chain variable region as set forth in SEQ ID NO: 298; (5) a light chain variable region as set forth in SEQ ID NO: 290 and a heavy chain variable region as set forth in SEQ ID NO:

A heavy chain variable region described by SEQ ID NO: 302; (6) a light chain variable region described by SEQ ID NO: 288 and a heavy chain variable region described by SEQ ID NO: 300; (7) a light chain variable region described by SEQ ID NO: 289 and a heavy chain variable region described by SEQ ID NO: 301; (8) a light chain variable region described by SEQ ID NO: 290 and a heavy chain variable region described by SEQ ID NO: 301; (9) a light chain variable region described by SEQ ID NO: 291 and a heavy chain variable region described by SEQ ID NO: 301; (10) a light chain variable region described by SEQ ID NO: 293 and a heavy chain variable region described by SEQ ID NO: 301; (11) a light chain variable region described by SEQ ID NO: 288 and a heavy chain variable region described by SEQ ID NO: 299; (12) a light chain variable region described by SEQ ID NO: 11 and a heavy chain variable region described by SEQ ID NO: 303; (13) a light chain variable region described by SEQ ID NO: 11 and a heavy chain variable region described by SEQ ID NO: 304 Variable region; (14) a light chain variable region as set forth in SEQ ID NO: 11 and a heavy chain variable region as set forth in SEQ ID NO: 307; (15) a light chain variable region as set forth in SEQ ID NO: 292 and a heavy chain variable region as set forth in SEQ ID NO: 303; (16) a light chain variable region as set forth in SEQ ID NO: 290 and a heavy chain variable region as set forth in SEQ ID NO: 307; (17) a light chain variable region as set forth in SEQ ID NO: 288 and a heavy chain variable region as set forth in SEQ ID NO: 305; (18) a light chain variable region as set forth in SEQ ID NO: 289 and a heavy chain variable region as set forth in SEQ ID NO: 306; (19) a light chain variable region as set forth in SEQ ID NO: 290 and a heavy chain variable region as set forth in SEQ ID NO: 306; (20) a light chain variable region as set forth in SEQ ID NO: 291 and a heavy chain variable region as set forth in SEQ ID NO: 306; (21) a light chain variable region as set forth in SEQ ID NO: 293 and a heavy chain variable region as set forth in SEQ ID NO: 306; (22) a light chain variable region as set forth in SEQ ID NO: 288 and a heavy chain variable region as set forth in SEQ ID NO: 304; (23) a light chain variable region as set forth in SEQ ID NO: 11 and a heavy chain variable region as set forth in SEQ ID NO: 308; (24) a light chain variable region as set forth in SEQ ID NO: 11 and a heavy chain variable region as set forth in SEQ ID NO: 309; (25) a light chain variable region as set forth in SEQ ID NO: 11 and a heavy chain variable region as set forth in SEQ ID NO: 312; (26) a light chain variable region as set forth in SEQ ID NO: 292 and a heavy chain variable region as set forth in SEQ ID NO: 308; (27) a light chain variable region as set forth in SEQ ID NO: 290 and a heavy chain variable region as set forth in SEQ ID NO: 312; (28) a light chain variable region as set forth in SEQ ID NO: 288 and a heavy chain variable region as set forth in SEQ ID NO: 310; (29) a light chain variable region as set forth in SEQ ID NO: 289 and a heavy chain variable region as set forth in SEQ ID NO: 311; (30) a light chain variable region described by SEQ ID NO: 290 and a heavy chain variable region described by SEQ ID NO: 311; (31) a light chain variable region described by SEQ ID NO: 291 and a heavy chain variable region described by SEQ ID NO: 311; (32) a sequence number A light chain variable region described as 293 and a heavy chain variable region described as SEQ ID NO: 311; or (33) a light chain variable region described as SEQ ID NO: 288 and a heavy chain variable region described as SEQ ID NO: 309.

[3] In any one of the above [1] to [29], the antibody or antigen-binding fragment thereof may include a light chain variable region comprising an amino acid sequence set forth in SEQ ID NO: 11 and a heavy chain variable region comprising an amino acid sequence set forth in SEQ ID NO: 308.

[31] In any one of [1] to [3] above, the antibody or antigen-binding fragment thereof and at least one of drug 1 and drug 2 may be administered simultaneously or separately.

[32] In any one of the above [1] to [31], the antibody or antigen-binding fragment thereof and at least one of drug 1 and drug 2 may be administered sequentially.

[33] In any one of the above [1] to [32], the pharmaceutical composition may reduce the size of a tumor.

[34] In any one of the above [1] to [33], the pharmaceutical composition may increase at least one selected from the group consisting of progression-free survival (PFS), overall survival (OS), complete response rate, and partial response rate of the subject.

[35] In any one of the above [1] to [34], the antibody or antigen-binding fragment thereof comprises a light chain variable region comprising a light chain CDR1 as set forth in SEQ ID NO: 1; a light chain CDR2 as set forth in SEQ ID NO: 2; a light chain CDR3 as set forth in SEQ ID NO: 3, and a light chain variable region comprising a light chain CDR1 as set forth in SEQ ID NO: 4. A heavy chain variable region comprising a heavy chain CDR1 described in SEQ ID NO: 5; a heavy chain CDR2 described in SEQ ID NO: 5; and a heavy chain CDR3 described in SEQ ID NO: 6, wherein (B) is the drug 1, and the cancer may be gastric cancer.

[36] In any one of the above [1] to [34], the antibody or antigen-binding fragment thereof comprises a light chain variable region comprising a light chain CDR1 as set forth in SEQ ID NO: 1; a light chain CDR2 as set forth in SEQ ID NO: 2; a light chain CDR3 as set forth in SEQ ID NO: 3, and a heavy chain variable region comprising a heavy chain CDR1 as set forth in SEQ ID NO: 4; a heavy chain CDR2 as set forth in SEQ ID NO: 5; and a heavy chain CDR3 as set forth in SEQ ID NO: 6, wherein (B) is the drug 2, and the cancer may be gastric cancer.

[37] In any one of the above [1] to [36], the antibody or antigen-binding fragment thereof may include a light chain variable region described in SEQ ID NO: 11 and a heavy chain variable region described in SEQ ID NO: 308.

[38] A pharmaceutical composition for preventing or treating cancer according to the present invention comprises, as an active ingredient, an antibody or an antigen-binding fragment thereof that specifically binds to hepatocyte growth factor receptor (c-Met) that binds to at least one epitope region described by an amino acid sequence selected from the group consisting of (A) SEQ ID NO: 313, SEQ ID NO: 314, SEQ ID NO: 315, and SEQ ID NO: 316, and the pharmaceutical composition may be administered to a subject in combination with (B) at least one of (1) Drug 1 and (2) Drug 2.

[39] The method for preventing or treating cancer according to the present invention comprises administering to an individual in need of prevention or treatment of cancer (A) SEQ ID NO: 313, SEQ ID NO: 314, SEQ ID NO: 315 and SEQ ID NO: A method for treating hepatocyte growth factor receptor (c-Met) comprising administering an antibody or antigen-binding fragment thereof that specifically binds to hepatocyte growth factor receptor (c-Met) that binds to at least one epitope region described by an amino acid sequence selected from the group consisting of 316, wherein the antibody or antigen-binding fragment thereof may be administered in combination with at least one of (1) drug 1 and (2) drug 2.

[4] The use of an antibody or antigen-binding fragment thereof that specifically binds to hepatocyte growth factor receptor (c-Met) that binds to at least one epitope region described by an amino acid sequence selected from the group consisting of (A) SEQ ID NO: 313, SEQ ID NO: 314, SEQ ID NO: 315, and SEQ ID NO: 316 according to the present invention for preventing or treating cancer may be such that the antibody or antigen-binding fragment thereof is administered in combination with (B) at least one of (1) Drug 1 and (2) Drug 2.

[41] The use of an antibody or antigen-binding fragment thereof that specifically binds to hepatocyte growth factor receptor (c-Met) that binds to at least one epitope region described by an amino acid sequence selected from the group consisting of (A) SEQ ID NO: 313, SEQ ID NO: 314, SEQ ID NO: 315 and SEQ ID NO: 316 in the manufacture of a medicament for use in preventing or treating cancer according to the present invention may be such that the antibody or antibody fragment is administered in combination with (B) at least one of (1) Drug 1 and (2) Drug 2.

[42] In any one of the above [38] to [41], (i) (A) a liver cell growth factor that binds to at least one epitope region described by an amino acid sequence selected from the group consisting of SEQ ID NO: 313, SEQ ID NO: 314, SEQ ID NO: 315 and SEQ ID NO: 316 An antibody or antigen-binding fragment thereof that specifically binds to factor receptor (c-Met), (ii) (B) drug 1, drug 2 or (iii) cancer may be substantially the same as described in [1] to [39] above, unless contradictory.

【Effect of the invention】 The combination therapy of the anti-c-Met antibody and the second anticancer agent of the present invention has excellent anticancer activity and can kill cancer cells by exhibiting a synergistic effect even in cancers that are difficult to overcome due to drug resistance, so it can be very usefully used for the prevention and treatment of cancer.

【Brief description of the drawing】 Figure 1 is a schematic diagram of a vector expressing a separate transcript for displaying the inside of the cell. Figure 2 is a result of analyzing the epitope of the antibody using hydrogen-deuterium exchange mass spectrometry (HDX-MS) and showing the result as a 3D structure. Figure 3 is a result of confirming the anticancer activity of the combined treatment of CM01 and SN-38 in the Hs746T cell line. Figure 4 is a result of confirming the anticancer activity of the combined treatment of CM01 and SN-38 in the SNU-719 cell line. Figure 5 is a result of confirming the anticancer activity of the combined treatment of CM01 and SN-38 in the MKN-45 cell line. Results. Figure 6 shows the results of measuring the anticancer effect of combined administration of CM01 and irinotecan in an MKN-45 cell xenograft model. ns: not significant.

【Best mode for carrying out the invention】 This is specifically described as follows. Meanwhile, each description and embodiment disclosed in the present invention can also be applied to each other description and embodiment. That is, all combinations of various elements disclosed in the present invention fall within the scope of the present invention. In addition, the scope of the present invention should not be considered limited by the specific descriptions described below. One aspect of the present invention for achieving the above object is a combination comprising (A) an antibody or an antigen-binding fragment thereof that specifically binds to hepatocyte growth factor receptor (c-Met) that binds to at least one epitope region described by an amino acid sequence selected from the group consisting of SEQ ID NO: 313, SEQ ID NO: 314, SEQ ID NO: 315, and SEQ ID NO: 316, and (B) at least one selected from the group consisting of (1) Drug 1, (2) Drug 2, (3) Drug 3, and (4) Drug 4 as an active ingredient.

(1) Drug 1: SN-38 represented by the following chemical formula 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof

[Chemical Formula 1]

(2) Drug 2: Irinotecan represented by the following chemical formula 2, an optical isomer thereof, or a pharmaceutically acceptable salt thereof

상기 조합물을 이용한 병용 요법은 우수한 항암 활성을 가지며, 약물 저항성을 가져 극복이 어려운 암에서도 시너지 효과를 나타내어 암 세포를 사멸시킬 수 있으므로 암의 예방 및 치료용도로 매우 유용하게 사용될 수 있다. 본 발명의 다른 하나의 양태는, (A) 서열번호 313, 서열번호 314, 서열번호 315 및 서열번호 316으로 이루어진 군으로부터 선택되는 아미노산 서열로 기재된 어느 하나 이상의 에피토프 영역에 결합하는 간 세포 성장 인자 수용체 (c- Met)에 특이적으로 결합하는 항체 또는 이의 항원 결합 단편, 및 (B) 상기 (1) 약물 1 및 (2) 약물 2 중 적어도 하나를 유효성분으로 포함하는, 암의 예방 또는 치료용 약학적 조성물이다. 본 발명의 또 다른 하나의 양태는, (A) 서열번호 313, 서열번호 314, 서열번호 315 및 서열번호 316으로 이루어진 군으로부터 선택되는 아미노산 서열로 기재된 어느 하나 이상의 에피토프 영역에 결합하는 간 세포 성장 인자 수용체 (c- Met)에 특이적으로 결합하는 항체 또는 이의 항원 결합 단편을 유효성분으로 포함하는 암의 예방 또는 치료용 약학적 조성물로서, 상기 약학적 조성물은 (B) 상기 (1) 약물 1 및 (2) 약물 2중 적어도 하나와 개체에 병용 투여된다. 본 발명의 또 다른 하나의 양태는, 암의 예방 또는 치료를 필요로 하는 개체에게 (A) 서열번호 313, 서열번호 314, 서열번호 315 및 서열번호 316으로 이루어진 군으로부터 선택되는 아미노산 서열로 기재된 어느 하나 이상의 에피토프 영역에 결합하는 간 세포 성장 인자 수용체 (c- Met)에 특이적으로 결합하는 항체 또는 이의 항원 결합 단편을 투여하는 단계를 포함하는 암의 예방 또는 치료 방법으로서, 상기 항체 또는 이의 항원 결합 단편은 (B) 상기 (1) 약물 1 및 (2) 약물 2중 적어도 하나와 병용 투여된다. 본 발명의 또 다른 하나의 양태는, (A) 서열번호 313, 서열번호 314, 서열번호 315 및 서열번호 316으로 이루어진 군으로부터 선택되는 아미노산 서열로 기재된 어느 하나 이상의 에피토프 영역에 결합하는 간 세포 성장 인자 수용체 (c- Met)에 특이적으로 결합하는 항체 또는 이의 항원 결합 단편의 암을 예방 또는 치료하는 용도로서, 상기 항체 또는 이의 항원 결합 단편은 (B) 상기 (1) 약물 1 및 (2) 약물 2중 적어도 하나와 병용 투여된다. 본 발명의 또 다른 하나의 양태는, 암을 예방 또는 치료하는데 사용하기 위한 약제의 제조에 있어서의 (A) 서열번호 313, 서열번호 314, 서열번호 315 및 서열번호 316으로 이루어진 군으로부터 선택되는 아미노산 서열로 기재된 어느 하나 이상의 에피토프 영역에 결합하는 간 세포 성장 인자 수용체 (c- Met)에 특이적으로 결합하는 항체 또는 이의 항원 결합 단편의 용도로서, 상기 항체 또는 항체 단편은 (B) 상기 (1) 약물 1 및 (2) 약물 2중 적어도 하나와 병용 투여된다. 본 발명의 조합물, 약학적 조성물, 또는 c- Met에 특이적으로 결합하는 항체 또는 이의 항원 결합 단편의 용도 및 이를 이용한 암을 예방 또는 치료하는 방법에 있어서, (i) (A) 서열번호 313, 서열번호 314, 서열번호 315 및 서열번호 316으로 이루어진 군으로부터 선택되는 아미노산 서열로 기재된 어느 하나 이상의 에피토프 영역에 결합하는 간 세포 성장 인자 수용체 (c- Met)에 특이적으로 결합하는 항체 또는 이의 항원 결합 단편, (ii) (B) 약물 1, 약물 2 또는 (iii) 암은 모순되지 않는 한 하기 기술하는 바에 의해 모두설명될 수 있다. [Chemical Formula 2]

Combination therapy using the above combination has excellent anticancer activity and can kill cancer cells by showing a synergistic effect even in cancers that are difficult to overcome due to drug resistance, so it can be very usefully used for the prevention and treatment of cancer. Another aspect of the present invention is (A) an antibody or an antigen-binding fragment thereof that specifically binds to hepatocyte growth factor receptor (c-Met) that binds to at least one epitope region described in an amino acid sequence selected from the group consisting of SEQ ID NO: 313, SEQ ID NO: 314, SEQ ID NO: 315, and SEQ ID NO: 316, and (B) the (1) drug 1 and (2) A pharmaceutical composition for preventing or treating cancer, comprising at least one of drug 2 as an active ingredient. Another aspect of the present invention is a pharmaceutical composition for preventing or treating cancer, comprising as an active ingredient (A) an antibody or antigen-binding fragment thereof that specifically binds to hepatocyte growth factor receptor (c-Met) that binds to at least one epitope region described in an amino acid sequence selected from the group consisting of SEQ ID NO: 313, SEQ ID NO: 314, SEQ ID NO: 315, and SEQ ID NO: 316, wherein the pharmaceutical composition is administered to a subject in combination with (B) at least one of (1) drug 1 and (2) drug 2. Another aspect of the present invention is a method for preventing or treating cancer, comprising the step of administering to a subject in need of prevention or treatment of cancer (A) an antibody or antigen-binding fragment thereof that specifically binds to hepatocyte growth factor receptor (c-Met) that binds to at least one epitope region described in an amino acid sequence selected from the group consisting of SEQ ID NO: 313, SEQ ID NO: 314, SEQ ID NO: 315, and SEQ ID NO: 316, wherein the antibody or antigen-binding fragment thereof is administered in combination with (B) at least one of (1) drug 1 and (2) drug 2. Another aspect of the present invention is a use of an antibody or antigen-binding fragment thereof that specifically binds to hepatocyte growth factor receptor (c-Met) that binds to at least one epitope region described by an amino acid sequence selected from the group consisting of (A) SEQ ID NO: 313, SEQ ID NO: 314, SEQ ID NO: 315 and SEQ ID NO: 316, for preventing or treating cancer, wherein the antibody or antigen-binding fragment thereof comprises (B) (1) drug 1 And (2) at least one of drug 2 is administered in combination. Another aspect of the present invention is the use of an antibody or antigen-binding fragment thereof that specifically binds to hepatocyte growth factor receptor (c-Met) that binds to at least one epitope region as described in an amino acid sequence selected from the group consisting of (A) SEQ ID NO: 313, SEQ ID NO: 314, SEQ ID NO: 315 and SEQ ID NO: 316 in the manufacture of a medicament for use in preventing or treating cancer, wherein the antibody or antibody fragment is administered in combination with (B) at least one of (1) drug 1 and (2) drug 2. In the use of the combination, pharmaceutical composition, or antibody or antigen-binding fragment thereof that specifically binds to c-Met of the present invention and the method for preventing or treating cancer using the same, (i) (A) an antibody or antigen-binding fragment thereof that specifically binds to hepatocyte growth factor receptor (c-Met) that binds to at least one epitope region described by an amino acid sequence selected from the group consisting of SEQ ID NO: 313, SEQ ID NO: 314, SEQ ID NO: 315 and SEQ ID NO: 316, (ii) (B) drug 1, drug 2 or (iii) cancer may all be described by the following unless contradictory.

I. Antibody or antigen-binding fragment thereof that specifically binds to hepatocyte growth factor receptor (c-Met) The antibody or antigen-binding fragment thereof of the present invention binds to hepatocyte growth factor receptor (c-

It binds specifically to Met and is administered in combination with a topoisomerase inhibitor. Since it exhibits a synergistic effect, it can be very useful in preventing and/or treating cancer. In the present invention, the term "antibody" refers to a protein molecule that acts as a ligand that specifically recognizes an antigen, including an immunoglobulin molecule that immunologically has reactivity with a specific antigen, and includes polyclonal antibodies, monoclonal antibodies, and whole antibodies. The term also includes chimeric antibodies and bivalent or bispecific molecules, diabodies, triabodies, and tetrabodies. The term further includes single-chain antibodies, scabs, derivatives of antibody constant regions, and artificial antibodies based on protein scaffolds that have binding function to FcRn. A whole antibody has a structure having two full-length light chains and two full-length heavy chains, each light chain being linked to a heavy chain by a disulfide bond. The above whole antibody includes IgA, IgD, IgE, IgM and IgG, and IgG includes subtypes such as IgG1, IgG2, IgG3 and IgG4. In the present invention, the terms "fragment", "antibody fragment" and "antigen-binding fragment" are used interchangeably to refer to any fragment of the antibody of the present invention that retains the antigen-binding function of the antibody. Exemplary antigen-binding fragments include, but are not limited to, Fab, Fab', F(ab')2, Fd, dsFv and scFv. The Fab has one antigen-binding site in a structure having variable regions of light and heavy chains, constant region of light chain and first constant region (CH1 domain) of heavy chain. An antigen-binding fragment of an antibody molecule or antibody fragment means a fragment that retains the antigen-binding function, and Fab' has one or more cysteine residues at the C-terminus of the heavy chain CH1 domain. It differs from Fab in that it has a hinge region that includes a disulfide bond. F(ab')2 antibodies are generated when cysteine residues in the hinge region of Fab' form a disulfide bond. Fd refers to the heavy chain portion included in the Fab fragment. Fv (variable fragment) refers to the smallest antibody fragment that has only the heavy chain variable region and the light chain variable region. Double disulfide Fv (dsFv) has the heavy chain variable region and the light chain variable region linked by a disulfide bond, and single-chain Fv (scFv) can form a dimer-like structure by covalently linking the heavy chain variable region and the single chain variable region through a peptide linker or directly linking them at the C-terminus. Although not limited thereto, such antibody fragments can be obtained using proteolytic enzymes (for example, restriction digestion of whole antibodies with papain yields Fab fragments and digestion with pepsin yields F(ab') ₂ fragments), or produced using genetic recombination techniques. c-Met (Hepatocyte growth factor receptor) is a type of RTK (Receptor tyrosine kinase) and a cell surface receptor for HGF/SF (hepatocyte growth factor, also known as scatter factor) (Laird AD et al. , Expert . Opin. Invest ig. Drugs 12：51-64 (2003)). Abnormal c-Met activation by HGF is one of the representative mechanisms of tumorigenesis and is known to be related to tumor proliferation, inhibition of apoptosis, formation of new blood vessels, invasion, and metastasis (Bottaro DP et al., Science 251: 802-804 (1991), Day RM et al., Oncogene 18: 3399-3406 (1999)). In addition, abnormal c-Met activation due to mutation and amplification of c-Met is associated with lung cancer, colon cancer, head and neck cancer, stomach cancer, and breast cancer. c-Met is associated with various cancers, and has been reported to be associated with increased tumor aggressiveness and poor prognosis (Lefebvre J et al., FASEB J 26: 1387-1399 (2012), Liu X et al., Trends Mol Med 16: 37—45 (2010), Smo 1 en GA et al., Proc Nat 1 Acad Sci USA 103: 2316-2321 (2006), Foveau B et al., Mol Biol Cell 20: 2495-2507 (2009)). Therefore, c-Met is attracting attention as a target antigen for treating various cancers, and various approaches are being attempted to inhibit the expression and activity of c-Met. In the present invention, the c-Met refers to a c-Met protein naturally present in a subject to be treated, for example, a cancer patient, and this is well known to those skilled in the art. The c-Met is activated by HGF and contributes to tumorigenesis, and even if some of the sequences are modified, that is, added, deleted, or substituted, it all falls within the scope of the present invention, and may be, for example, c-Met represented by the amino acid sequence set forth in SEQ ID NO: 317, but is not limited thereto. Specifically, in the present invention, the anti-c-Met antibody may be an antibody comprising a light chain variable region including a light chain CDR1 set forth in SEQ ID NO: 1; a light chain CDR2 set forth in SEQ ID NO: 2; and a light chain CDR3 set forth in SEQ ID NO: 3, and a heavy chain variable region including a heavy chain CDR1 set forth in SEQ ID NO: 4; a heavy chain CDR2 set forth in SEQ ID NO: 5; and a heavy chain CDR3 set forth in SEQ ID NO: 6, or an affinity-optimized antibody thereof. In the present invention, the term "heavy chain" includes a full-length heavy chain and fragments thereof, including a variable region domain VH comprising an amino acid sequence having sufficient variable region sequence to confer specificity to an antigen and three constant region domains CHI, CH2 and CH3. may include. In addition, the term "light chain" in the present invention may include both a full-length light chain and a fragment thereof, including a variable region domain VL and a constant region domain CL, which include an amino acid sequence having sufficient variable region sequence to confer specificity to an antigen. In the present invention, the antibody may include both a mouse antibody produced from a mouse and a variant in which a part of the amino acid sequence of the parent antibody is substituted, added and/or deleted in order to improve the affinity, immunogenicity, etc. of the antibody therefrom. The variant is not limited thereto, but may include, for example, a chimeric antibody, a humanized antibody, an affinity-optimized antibody, etc. In the present invention, the variant comprehensively refers to an antibody that includes the same CDR as the parent antibody, or an antibody in which a part of the parent antibody CDR amino acid sequence is mutated (substituted, added or deleted) under the condition of targeting the same epitope. Such variants can be appropriately adjusted by those skilled in the art to improve the affinity and immunogenicity of the antibody within the range in which the binding ability to the same epitope is maintained. In other words, the antibody or antigen-binding fragment thereof of the present invention may include not only the sequence of the anti-c-Met antibody described herein but also a biological equivalent thereof within the range in which it can specifically recognize c-Met. For example, additional changes may be made to the amino acid sequence of the antibody to further improve the binding affinity and/or other biological properties of the antibody. Such modifications include, for example, deletion, insertion and/or substitution of amino acid sequence residues of the antibody. Such amino acid Mutations are made based on the relative similarity of amino acid side chain substituents, such as hydrophobicity, hydrophilicity, charge, size, etc. Analysis of the size, shape, and type of amino acid side chain substituents reveals that arginine, lysine, and histidine are all positively charged residues; alanine, glycine, and serine have similar sizes; and phenylalanine, tryptophan, and tyrosine have similar shapes. Therefore, based on these considerations, arginine, lysine, and histidine; alanine, glycine, and serine; and phenylalanine, tryptophan, and tyrosine can be considered biologically functional equivalents. The term "chimeric antibody" as used herein refers to an antibody obtained by recombining the variable region of a mouse antibody and the constant region of a human antibody, and is an antibody whose immune response is significantly improved compared to a mouse antibody. In the present invention, the term "humanized antibody" refers to an antibody in which the protein sequence of an antibody derived from a non-human species is modified to resemble an antibody variant naturally produced in humans. For example, the humanized antibody can be produced by recombining a mouse-derived CDR with a human antibody-derived FR to produce a humanized variable region, and recombining this with a desired human antibody constant region. However, since the affinity of the humanized antibody is reduced when only CDR grafting is performed, the affinity of several important FR amino acid residues that are thought to affect the three-dimensional structure of the CDR can be increased to the same level as the original mouse antibody by affinizing them with those of a mouse antibody. The term "affinity-optimized antibody" in the present invention refers to an antibody in which a part of the CDR sequence of a specific antibody is optimized. Refers to an antibody having improved binding affinity for the antigen while binding to the same antigen epitope as the specific antibody, as a substituted, added, or deleted variant. Specifically, the affinity-optimized antibody of the present invention refers to a variant antibody that binds to the same epitope as the antibody, comprising a light chain variable region comprising a light chain CDR1 as set forth in SEQ ID NO: 1; a light chain CDR2 as set forth in SEQ ID NO: 2; a light chain CDR3 as set forth in SEQ ID NO: 3; and a heavy chain variable region comprising a heavy chain CDR1 as set forth in SEQ ID NO: 4, a heavy chain CDR2 as set forth in SEQ ID NO: 5; and a heavy chain CDR3 as set forth in SEQ ID NO: 6. A person skilled in the art can produce the affinity-optimized antibody using a known technique based on the specified light chain and heavy chain CDR sequences. For example, the affinity-optimized antibody of the present invention can be produced through phage display. As used herein, the term "phage display" refers to a technique for displaying a variant polypeptide as a fusion protein with at least a portion of a coat protein on the surface of a phage, for example, a filamentous phage particle. The utility of phage display lies in the fact that it can rapidly and efficiently sort out sequences that bind with high affinity to a target antigen from a large library of randomized protein variants. Displaying peptide and protein libraries on phage has been used to screen millions of polypeptides to identify polypeptides with specific binding properties. In one embodiment of the present invention, the antibody may be an antibody comprising a light chain variable region as set forth in SEQ ID NO: 7 and a heavy chain variable region as set forth in SEQ ID NO: 8. In one embodiment, the antibody may comprise a light chain variable region encoded by nucleotides as set forth in SEQ ID NO: 9 and It may be an antibody comprising a heavy chain variable region encoded by the nucleotides set forth in SEQ ID NO: 10, but is not limited thereto. According to a specific embodiment of the present invention, a group of hybridoma cells was secured from a mouse using a human c-Met Sema domain/Fc fusion protein as an antigen, and screening was performed from these using an ELISA analysis method using a c-Met/His fusion protein as an antigen, thereby selecting an anti-c-Met antibody that specifically binds to c-Met. In another embodiment of the present invention, the antibody may comprise (a) a light chain variable region set forth in SEQ ID NO: 11 and a heavy chain variable region set forth in SEQ ID NO: 13; or (b) a light chain variable region set forth in SEQ ID NO: 12 and a heavy chain variable region set forth in SEQ ID NO: 14. As an example, the antibody may comprise (a) a light chain variable region encoded by the nucleotides set forth in SEQ ID NO: 15 and a heavy chain variable region encoded by the nucleotides set forth in SEQ ID NO: 17; Or (b) may comprise a light chain variable region encoded by a nucleotide set forth in SEQ ID NO: 16 and a heavy chain variable region encoded by a nucleotide set forth in SEQ ID NO: 18, but is not limited thereto. In addition, the antibody may comprise a hinge region set forth in any one of SEQ ID NOs: 19 to 26. In another embodiment of the present invention, but is not limited thereto, an affinity-optimized antibody for the antibody comprises a light chain variable region comprising a light chain CDR1 set forth in SEQ ID NO: 1; a light chain CDR2 set forth in SEQ ID NO: 2; and a light chain CDR3 set forth in SEQ ID NO: 3, and An antibody comprising a heavy chain variable region comprising a heavy chain CDR1 as set forth in SEQ ID NO: 4, a heavy chain CDR2 as set forth in SEQ ID NO: 5; and a heavy chain CDR3 as set forth in SEQ ID NO: 6, wherein at least one amino acid sequence is substituted, wherein (i) the E at the first position of the light chain CDR1 is A, E, K, L, N, R, S, V or W; the A at the second position is C, G, I, P, S, T or V; and the E at the third position is

G, M, N, P, Q, R, S or T; E at the 4th position is A, D, F, G, H, K, M, Q, R, S, T or V; 凡 at the 5th position is A, D, E, G, K, L, P, Q, R, S, T or V; I at the 6th position is A, F, L, M, Q, R, S, T or V; Y at the 7th position is F, H, R or V; or G at the 8th position is replaced with D, F, H, M, N, R, S, T or V; ( i i ) the 1st position of the light chain CDR2

G is D, F, H, K, P, Q, S, V or Y; T in the 3rd position is Q; or N in the 4th position is

Substitution with G; ( i i i ) The 1st position of the light chain CDR3 is E, G, I, M or N; The 2nd position is A, D, E, H, L, Q, S or T; The 3rd position is I, L, M, N, Q, S or T; The 4th position is F, H, I, M, R, S, V, W or Y; The 5th position is S, C, D, E, F, G, H, K, L, N, Q, R, T, V or Y; The 6th position is ▽} D, E, F, G, H, I, L, M, N, P, Q, R, T, V or Y; The 7th position is P, A, D, E, G, N, Q, S or V; The 8th position is Y

E, F, L, M or Q; or T at the 9th position is replaced with D, F, G, I, L, N, S, V, W or Y; (iv) Di at the 1st position of heavy chain CDR1 is replaced with G or Q; Pentavalent Q at the 2nd position; or I at the 4th position is replaced with A or Y; (v) F at the 3rd position of heavy chain CDR2 is replaced with D, E, W or Y; G at the 5th position is replaced with D, H or Y; E at the 6th position is replaced with F, P, W or Y; G at the 7th position is replaced with A, F, L, N or T; N at the 8th position is replaced with F, P, S, T or Y; T at the 9th position is replaced with A, D, E, F, G, H, L, P, S or V; H at the 10th position is replaced with A, D, F, M, R, S, T, V, W or Y; F in the 11th position is G, H, I, L, M, N, P, Q, V or Y; F in the 12th position is G, H, I, L, M, N, P, Q, V or Y;

S is A, D, G, H, I, L, P, T or V; A in the 13th position is D, E, F, G, H, I, K, L,

M, P, R, S, T, V or Y; R in the 14th position is A, E, G, H, L, N, P, Q, S, W or

Y； F in the 15th position is D, E, G, L, M, P, R, S, V or W； O in the 16th position is A, E,

F, G, H, L, R, S, T, V or Y; or G in the 17th position is E, F, H, L, M, N, P, Q,

R, S, T, V or W; or (vi) G at position 1 of heavy chain CDR3 is replaced by E, F, H, N,

Q, V or W; 日 in the 2nd position is E; Y in the 3rd position is L, Q, T or V; Y in the 4th position is

G is W; F at the 5th position is L or Y; L at the 6th position is Q, S or Y; or O at the 7th position is C, L, M, N or Y, wherein the light chain CDR1 is 0 to 5, light chain

CDR2 is 0 to 1, light chain CDR3 is 0 to 7, heavy chain CDR1 is 0 to 1, heavy chain

CDR2 may contain 0 to 11 substitutions, and heavy chain CDR3 may contain 0 to 6 substitutions. In another embodiment of the present invention, the affinity-optimized antibody specifically comprises a light chain variable region comprising a light chain CDR1 as described in any one of SEQ ID NO: 1 and SEQ ID NOs: 211 to 250; a light chain CDR2 as described in any one of SEQ ID NO: 2, SEQ ID NOs: 164 to 172, SEQ ID NO: 209 and SEQ ID NO: 210; a light chain CDR3 as described in any one of SEQ ID NO: 3, SEQ ID NOs: 124 to 163, SEQ ID NOs: 173 to 208, and SEQ ID NOs: 251 to 283; and a heavy chain CDR1 as described in any one of SEQ ID NO: 4 and SEQ ID NOs: 90 to 94; a heavy chain CDR2 as described in any one of SEQ ID NO: 5, SEQ ID NOs: 36 to 45, SEQ ID NOs: 54 to 89, and SEQ ID NOs: 100 to 123; and a heavy chain CDR1 as described in any one of SEQ ID NO: 6, SEQ ID NOs: 46 to 53, and SEQ ID NOs: 95 to 99. Heavy chain containing heavy chain CDR3 It may comprise a variable region, and more specifically, it may comprise a light chain variable region as set forth in any one of SEQ ID NOs: 11 and 288 to 293 and a heavy chain variable region as set forth in any one of SEQ ID NOs: 13 and 284 to 287, and more specifically,

(a) a light chain variable region described by SEQ ID NO: 11 and a heavy chain variable region described by SEQ ID NO: 284;

(b) a light chain variable region described by SEQ ID NO: 11 and a heavy chain variable region described by SEQ ID NO: 287;

(c) a light chain variable region as set forth in SEQ ID NO: 292 and a heavy chain variable region as set forth in SEQ ID NO: 13; (d) a light chain variable region as set forth in SEQ ID NO: 290 and a heavy chain variable region as set forth in SEQ ID NO: 287; (e) a light chain variable region as set forth in SEQ ID NO: 288 and a heavy chain variable region as set forth in SEQ ID NO: 285; (f) a light chain variable region as set forth in SEQ ID NO: 289 and a heavy chain variable region as set forth in SEQ ID NO: 286; (g) a light chain variable region as set forth in SEQ ID NO: 290 and a heavy chain variable region as set forth in SEQ ID NO: 286; (h) a light chain variable region as set forth in SEQ ID NO: 291 and a heavy chain variable region as set forth in SEQ ID NO: 286; (i) a light chain variable region as set forth in SEQ ID NO: 293 and a heavy chain variable region as set forth in SEQ ID NO: 286; or (j) a light chain variable region as set forth in SEQ ID NO: 288 and a heavy chain variable region as set forth in SEQ ID NO: 284, but is not limited thereto. In the present invention, the antibody may be an antibody or antigen-binding fragment thereof that specifically binds to epidermal growth factor receptor (EGFR) in addition to specifically binding to c-Met. That is, the antibody in the present invention may be a c-Met monospecific antibody or a dual specific antibody that simultaneously targets c-Met and EGFR.

EGFR (Epidermal growth factor receptor) is one of the ErbB tyrosine kinases. It is known to be abnormally activated in many epithelial cell tumors including non-small cell lung carcinoma, and to cause cell proliferation, invasion, metastasis, angiogenesis, and increase cell survival. In one embodiment, the bispecific antibody may be one in which an antibody or an antigen-binding fragment thereof that specifically binds to EGFR is linked to one end of the light chain or heavy chain of a c-Met specific antibody, for example, but not limited to, one linked to the C-terminus of the heavy chain. The binding fragment that specifically binds to EGFR may be Fab, Fab', F(ab') ₂ , or Fv. In one embodiment of the present invention, the Fv may be a scFv fragment, and the scFv fragment may be linked using a connector capable of linking the scFv fragment to one end of the light chain or heavy chain of a c-Met antibody. In one embodiment of the present invention, an antibody that specifically binds to EGFR was further produced by linking using a connector as set forth in SEQ ID NO: 294. The EGFR scFv fragment may be an EGFR scFv known in the art that can specifically bind to EGFR, such as Erbitux (cetuxidab), Vectibix (panitumudab), Portrazza (nesitumumab), or TheraCIM (nimotuzumab), but is not limited thereto. In one embodiment of the present invention, the EGFR scFv may be an Erbitux or Vectibix scFv fragment, and specifically, the Erbitux scFv may be a fragment of SEQ ID NO: 295 or SEQ ID NO: It may include an amino acid sequence described as SEQ ID NO: 296, and the Vectibix scFv may include an amino acid sequence described as SEQ ID NO: 297, but is not limited thereto. In one embodiment of the present invention, the antibody or antigen-binding fragment thereof may include an amino acid sequence described as SEQ ID NO:

11, and a light chain variable region as set forth in any one of SEQ ID NOs: 288 to 293; and a heavy chain variable region as set forth in any one of SEQ ID NOs: 298 to 312, and specifically, the antibody or antigen-binding fragment thereof may comprise (1) SEQ ID NOs:

A light chain variable region described as 11 and a heavy chain variable region described as SEQ ID NO: 298; (2) SEQ ID NO:

A light chain variable region described as 11 and a heavy chain variable region described as SEQ ID NO: 299; (3) SEQ ID NO:

A light chain variable region described as 11 and a heavy chain variable region described as SEQ ID NO: 302; (4) SEQ ID NO:

A light chain variable region described as SEQ ID NO: 292 and a heavy chain variable region described as SEQ ID NO: 298; (5) a light chain variable region described as SEQ ID NO: 290 and a heavy chain variable region described as SEQ ID NO: 302;

(6) a light chain variable region as set forth in SEQ ID NO: 288 and a heavy chain variable region as set forth in SEQ ID NO: 300; (7) a light chain variable region as set forth in SEQ ID NO: 289 and a heavy chain variable region as set forth in SEQ ID NO: 301; (8) a light chain variable region as set forth in SEQ ID NO: 290 and a heavy chain variable region as set forth in SEQ ID NO: 301; (9) a light chain variable region as set forth in SEQ ID NO: 291 and a heavy chain variable region as set forth in SEQ ID NO: 301; (10) a light chain variable region as set forth in SEQ ID NO: 293 and a heavy chain variable region as set forth in SEQ ID NO: 301; (11) a light chain variable region as set forth in SEQ ID NO: 288 and a heavy chain variable region as set forth in SEQ ID NO: 299; (12) a light chain variable region as set forth in SEQ ID NO: 11 and a heavy chain variable region as set forth in SEQ ID NO: 303; (13) a light chain variable region as set forth in SEQ ID NO: 11 and a heavy chain variable region as set forth in SEQ ID NO: 304; (14) Sequence number 11 The light chain variable region described and the heavy chain variable region described as SEQ ID NO: 307; (15) SEQ ID NO:

A light chain variable region described as 292 and a heavy chain variable region described as SEQ ID NO: 303; (16) A light chain variable region described as SEQ ID NO: 290 and a heavy chain variable region described as SEQ ID NO: 307;

(17) a light chain variable region as set forth in SEQ ID NO: 288 and a heavy chain variable region as set forth in SEQ ID NO: 305; (18) a light chain variable region as set forth in SEQ ID NO: 289 and a heavy chain variable region as set forth in SEQ ID NO: 306; (19) a light chain variable region as set forth in SEQ ID NO: 290 and a heavy chain variable region as set forth in SEQ ID NO: 306; (20) a light chain variable region as set forth in SEQ ID NO: 291 and a heavy chain variable region as set forth in SEQ ID NO:

A heavy chain variable region described by SEQ ID NO: 306; (21) a light chain variable region described by SEQ ID NO: 293 and a heavy chain variable region described by SEQ ID NO: 306; (22) a light chain variable region described by SEQ ID NO: 288 and a heavy chain variable region described by SEQ ID NO: 304; (23) a light chain variable region described by SEQ ID NO: 11 and a heavy chain variable region described by SEQ ID NO: 308; (24) a light chain variable region described by SEQ ID NO: 11 and a heavy chain variable region described by SEQ ID NO: 309; (25) a sequence

A light chain variable region as set forth in SEQ ID NO: 11 and a heavy chain variable region as set forth in SEQ ID NO: 312; (26) a light chain variable region as set forth in SEQ ID NO: 292 and a heavy chain variable region as set forth in SEQ ID NO: 308; (27) a light chain variable region as set forth in SEQ ID NO: 290 and a heavy chain variable region as set forth in SEQ ID NO: 312; (28) a light chain variable region as set forth in SEQ ID NO: 288 and a heavy chain variable region as set forth in SEQ ID NO: 310; (29) a light chain variable region as set forth in SEQ ID NO: 289 and a heavy chain variable region as set forth in SEQ ID NO: 311; (30) a light chain variable region as set forth in SEQ ID NO: 290 and a heavy chain variable region as set forth in SEQ ID NO: 311; (31) a light chain variable region as set forth in SEQ ID NO: 291 and a heavy chain variable region as set forth in SEQ ID NO: 311; (32) a light chain variable region as set forth in SEQ ID NO: 293 and a heavy chain variable region as set forth in SEQ ID NO: 311; Or (33) a light chain variable region as described in SEQ ID NO: 288 and a heavy chain variable region as described in SEQ ID NO: 309. And more specifically, the antibody or antigen-binding fragment thereof may include a light chain variable region comprising an amino acid sequence set forth in SEQ ID NO: 11 and a heavy chain variable region comprising an amino acid sequence set forth in SEQ ID NO: 308, but is not limited thereto. The antibody or antigen-binding fragment thereof of the present invention may bind to an epitope region set forth in an amino acid sequence selected from the group consisting of SEQ ID NO: 313, SEQ ID NO: 314, SEQ ID NO: 315 and/or SEQ ID NO: 316. An affinity-optimized antibody produced based on a specific antibody (reference antibody) is characterized by having high homology with the light and heavy chain CDR sequences of the variable region of the reference antibody and binding to the same epitope region as the reference antibody, and thus may share all biological characteristics such as binding site, specificity, and pharmacological mechanism and pharmacological effect caused by the antibody, and may exhibit a superior effect than the reference antibody in terms of binding affinity. The above epitope region means, for example, YVSKPGAQL (SEQ ID NO: 313) at positions 321 to 329 from the N-terminus in the reference c-Met antigen (SEQ ID NO: 317), IGASLNDDI (SEQ ID NO: 314) at positions 333 to 341, PIKYVND (SEQ ID NO: 315) at positions 366 to 372, and QVWSRSGPST (SEQ ID NO: 316) at positions 464 to 474, respectively, and even if the c-Met antigen sequence to which the antibody or antigen-binding fragment thereof binds includes some mutations (substitutions, additions, or deletions), or the binding site or sequence is somewhat different because the antigen to which it binds exists in the form of a fragment, precursor, or subtype of c-Met, a person skilled in the art will recognize the epitope sequence of the reference c-Met antigen. Based on the information, the position and sequence to which the antigen of the present invention or its antigen-binding fragment binds can be clearly specified. In a specific embodiment of the present invention, it was confirmed that the antibody hu8C4 x Vect ibix scFv of the present invention binds to four epitope regions of the human c-Met sema domain P chain: Y321 to L329 (SEQ ID NO: 313), 1333 to 1341 (SEQ ID NO: 314), P366 to D372 (SEQ ID NO: 315), and Q464 to S474 (SEQ ID NO: 316) (Fig. 2). Therefore, in one embodiment, the antibody of the present invention may bind to the epitope regions of SEQ ID NO: 313, SEQ ID NO: 314, SEQ ID NO: 315, and SEQ ID NO: 316, but is not limited thereto. The "antibody or antigen-binding fragment thereof that specifically binds to c-Met" of the present invention means one that binds to human c-Met with a _KD of 1 x 10' ⁷ M or less. The antibody or antigen-binding fragment may, for example, have a KD of 5 x 10' ⁸ M or less, a KD of 1 x 10' ⁸ M or less, a KD of 5 x 10'

⁹ M or less, or KD 1 x 10' ⁹ M or less, but is not limited thereto. The binding affinities of hu8C4, hu8C4 AH71 and hu8C4 x Vect ibix scFv (CM01) to c-Met ECD show KD values of 3.173 x 10' ⁰ , 9.993 x 10' ¹¹ and 2.78 x 10' 11, respectively, so that the antibodies or antigen-binding fragments thereof of the present invention have high binding affinity to the c-Met antigen. The antibodies or antigen-binding fragments thereof of the present invention have cross-reactivity to the c-Met antigen of the same cynomolgus monkey, but not to other animals. It does not bind to antigens derived from humans (e.g., rodents). In addition, the antibody or antigen-binding fragment thereof of the present invention does not bind to cell surface receptors other than c-Met. Therefore, it can be seen from the above results that the antibody or antigen-binding fragment thereof of the present invention exhibits binding specificity to c-Met antigens of humans and monkeys. The term "association constant (Kon)" used in the present invention refers to the association ratio of a specific antibody-antigen interaction, and the term "dissociation constant (K ₀ ff )" refers to the dissociation ratio of a specific antibody-antigen interaction. In addition, the term "affinity for antigen (KD)" used in the present invention refers to the ratio of Koff:Kon (i.e., Koff / Kon) expressed as a molar concentration (M). The KD value for an antibody can be measured using a method widely established in the art. For example, a method for measuring the KD value of an antibody may include, but is not limited to, surface plasmon resonance analysis using the BioCore™ system.

II. Second anticancer agent The antibody or antigen-binding fragment thereof that specifically binds to c-Met of the present invention can exhibit an unexpected synergistic effect in anticancer treatment by being administered in combination with a second anticancer agent. The second anticancer agent of the present invention is at least one of the following (1) drug 1 and (2) drug 2. As a specific example, the pharmaceutical composition of the present invention may comprise an antibody or antigen-binding fragment thereof that specifically binds to c-Met, and drug 1; or an antibody or antigen-binding fragment thereof that specifically binds to c-Met, and drug 2 as active ingredients. In the present invention, (1) drug 1 refers to SN-38 represented by the following chemical formula 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof:

본 발명에서 (2) 약물 2는 하기 화학식 2로 표시되는 이리노테칸[Chemical Formula 1]

In the present invention, (2) drug 2 is irinotecan represented by the following chemical formula 2.

(irinotecan), its optical isomers or pharmaceutically acceptable salts thereof:

본 발명에서 확인된 바와 같이 , 상기 약물 1과 약물 2는 단독으로 사용되는 경우에 비하여 본 발명의 c-Met 항체와 병용으로 사용되었을 때 예측하지 못한 현저한 시너지 효과를 나타낸다. 따라서 본 발명의 (A) 간 세포 성장 인자 수용체 (c- Met )에 특이적으로 결합하는 항체 또는 이의 항원 결합 단편 , 및 (B) (1) 약물 1 및 (2) 약물 2 중 적어도 하나를 유효성분으로 포함하는 조합물은 암의 예방 및/또는 치료에 매우 유용하게 사용될 수 있다. 본 발명에서 "약학적으로 허용 가능한"은 생리학적으로 허용되고 개체에게 투여될 때 , 통상적으로 위장 장애 , 현기증과 같은 알러지 반응 또는 이와 유사한 반응을 일으키지 않는 것을 의미할 수 있다. 본 발명의 약학적으로 허용 가능한 염은 당업계 통상의 기술자에게 공지된 통상적인 방법에 의해 제조될 수 있다. 본 발명의 약학적으로 허용 가능한 염은 약학적으로 허용되는 음이온을 함유하는 무독성 산부가염을 형성하는 산을 이용해 제조된 것이면 특별히 한정하지 않는다. 예를 들면 , 염산, 황산, 질산, 인산, 브롬화수소산, 요오드화수소산 등으로 제조된 무기산염 ; 타르타르산, 포름산, 시트르산, 아세트산, 트리클로로아세트산 , 트리플루오로아세트산 , 글루콘산 , 벤조산 , 락트산 , 푸마르산 , 말레인산 등으로 제조된 유기산염 , 메탄설폰산, 벤젠설폰산, p-톨루엔설폰산, 나프탈렌설폰산 등으로 제조된 설폰산염 등을 들 수 있으나, 이에 한정하지 않는다. 본 발명에 있어서 상기 약물 1 및 약물 2 중 어느 하나에 따른 화합물은 1 개 이상의 비대칭 탄소 중심을 가질 수 있으며 , 이에 따라 라세믹 혼합물을 포함하는 에난티오머 혼합물 , 단일의 에난티오머 (광학이성질체 ) , 부분입체이성질체 혼합물 및 단일의 부분입체이성질체로서 존재할 수 있다. 이러한 이성질체는 종래기술 , 예를 들어 관 크로마토그래피 또는 HPLC 등의 분할에 의해 분리가 가능하다. 또는 , 공지된 배열의 광학적으로 순수한 출발물질 및/또는 시약을 사용하여 입체특이적으로 합성할 수 있다. 구체적으로, 상기 입체 이성질체는 광학이성질체일 수 있다. [Chemical Formula 2]

As confirmed in the present invention, the above drugs 1 and 2 exhibit an unexpectedly remarkable synergistic effect when used in combination with the c-Met antibody of the present invention, compared to when used alone. Therefore, (A) an antibody or antigen-binding fragment thereof that specifically binds to hepatocyte growth factor receptor (c-Met), and (B) (1) A combination comprising at least one of drug 1 and (2) drug 2 as an active ingredient can be very useful for the prevention and/or treatment of cancer. In the present invention, "pharmaceutically acceptable" can mean physiologically acceptable and, when administered to a subject, does not typically cause an allergic reaction such as gastrointestinal disorder or dizziness or a similar reaction. The pharmaceutically acceptable salt of the present invention can be prepared by a conventional method known to those skilled in the art. The pharmaceutically acceptable salt of the present invention is not particularly limited as long as it is prepared using an acid that forms a non-toxic acid addition salt containing a pharmaceutically acceptable anion. For example, inorganic acid salts prepared with hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, hydrobromic acid, hydroiodic acid, etc.; Organic acid salts made of tartaric acid, formic acid, citric acid, acetic acid, trichloroacetic acid, trifluoroacetic acid, gluconic acid, benzoic acid, lactic acid, fumaric acid, maleic acid, etc., sulfonic acid salts made of methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, etc., but are not limited thereto. In the present invention, the compound according to any one of the drug 1 and drug 2 may have one or more asymmetric carbon centers, and thus may exist as an enantiomer mixture including a racemic mixture, a single enantiomer (optical isomer), a diastereomeric mixture, and a single diastereoisomer. Such isomers can be separated by resolution using conventional techniques, for example, column chromatography or HPLC. Alternatively, an optically pure starting material of a known configuration and/or It can be synthesized stereospecifically using a reagent. Specifically, the stereoisomer may be an optical isomer.

III. Pharmaceutical composition, use, and method for preventing or treating cancer using the same The pharmaceutical composition of the present invention comprises an antibody or an antigen-binding fragment thereof that specifically binds to c-Met, and is administered together with a second anticancer agent to induce cancer cell death, and thus can be usefully used for the treatment or prevention of cancer. In the present invention, the cancer may be a solid cancer or a blood cancer. For example, pseudomyxoma, intrahepatic cholangiocarcinoma, hepatoblastoma, liver cancer, thyroid cancer, colon cancer, testicular cancer, myelodysplastic syndrome, glioblastoma, oral cancer, lip cancer, mycosis fungoides, acute myeloid leukemia, acute lymphoblastic leukemia, basal cell carcinoma, ovarian epithelial cancer, ovarian germ cell cancer, male breast cancer, brain cancer, pituitary adenoma, multiple myeloma, gallbladder cancer, biliary tract cancer, colon cancer, chronic myeloid leukemia, chronic lymphocytic leukemia, retinoblastoma, choroidal melanoma, ampulla of Vater cancer, bladder cancer, peritoneal cancer, parathyroid cancer, adrenal cancer, mast cell tumor, paranasal sinus cancer, non-small cell lung cancer, tongue cancer, astrocytoma, small cell lung cancer, pediatric brain cancer, pediatric lymphoma, pediatric leukemia, small intestine cancer, meningioma, esophageal cancer, glioma, Renal pelvic cancer, kidney cancer, heart cancer, duodenal cancer, malignant soft tissue cancer, malignant bone cancer, malignant lymphoma, malignant mesothelioma, malignant melanoma, eye cancer, vulvar cancer, ureteral cancer, urethral cancer, cancer of unknown primary site, gastric lymphoma, stomach cancer, gastric carcinoid tumor, gastrointestinal stromal tumor, gastrointestinal stromal cancer, Wilms' cancer, breast cancer, sarcoma, penile cancer, pharyngeal cancer, gestational trophoblastic disease, cervical cancer, endometrial cancer, uterine sarcoma, prostate cancer, metastatic bone cancer, metastatic brain cancer, mediastinal cancer, rectal cancer, rectal carcinoid tumor, vaginal cancer, spinal cord cancer, It may be at least one selected from the group consisting of acoustic neuroma, pancreatic cancer, salivary gland cancer, Kaposi's sarcoma, Paget's disease, tonsillar cancer, squamous cell carcinoma, lung adenocarcinoma, lung cancer, lung squamous cell carcinoma, skin cancer, anal cancer, rhabdomyosarcoma, laryngeal cancer, pleural cancer, blood cancer, and thymic cancer, and specifically, at least one of gastric cancer, colon cancer, and esophageal cancer, and more specifically, gastric cancer, but is not limited thereto. In one embodiment of the present invention, the cancer may be primary, recurrent, metastatic, or advanced cancer. In one embodiment of the present invention, the cancer may be resistant to chemotherapy. In the present invention, the chemotherapy is fluorouracil, capecitabine, oxaliplatin, epirubicin, cisplatin, carboplatin, etoposide, doxorubicin, methotrexate, leucovorin.

( leucovorin), irinotecan, docetaxel, paclitaxel, trifluridine/tipiracil (trif luridine/tipiracil 1) or a combination thereof. In another embodiment of the present invention, the cancer may be a c-Met wild-type cancer or a cancer that overexpresses c-Met, amplifies c-Met, or activates c-Met. The antibody or antigen-binding fragment thereof and at least one of drug 1 and drug 2 may be administered simultaneously or separately. The pharmaceutical composition of the present invention comprises an antibody Or the antibody or antigen-binding fragment thereof and at least one of drug 1 and drug 2 may be included together as a single pharmaceutical composition, or may be included as separate compositions. The antibody or antigen-binding fragment thereof and at least one of drug 1 and drug 2 may be administered simultaneously or sequentially. In the present invention, the pharmaceutical composition may be one that reduces the size of a tumor. In addition, the pharmaceutical composition may be one that increases at least one selected from the group consisting of progression-free survival (PFS), overall survival (OS), complete response rate, and partial response rate of a subject. As a specific example, in the pharmaceutical composition of the present invention, the antibody or antigen-binding fragment thereof comprises a light chain variable region comprising a light chain CDR1 as set forth in SEQ ID NO: 1; a light chain CDR2 as set forth in SEQ ID NO: 2; a light chain CDR3 as set forth in SEQ ID NO: 3; and a heavy chain variable region comprising a heavy chain CDR1 as set forth in SEQ ID NO: 4; a heavy chain CDR2 as set forth in SEQ ID NO: 5; and a heavy chain CDR3 as set forth in SEQ ID NO: 6, wherein the second anticancer agent is drug 1, and the cancer may be gastric cancer. The gastric cancer may be a c-Met wild-type cancer or a cancer in which c-Met is amplified or activated by c-Met exon 14 skipping, but is not limited thereto. As another specific example, in the pharmaceutical composition of the present invention, the antibody or antigen-binding fragment thereof comprises a light chain variable region comprising a light chain CDR1 as set forth in SEQ ID NO: 1; a light chain CDR2 as set forth in SEQ ID NO: 2; a light chain CDR3 as set forth in SEQ ID NO: 3; and a heavy chain variable region comprising a heavy chain CDR1 as set forth in SEQ ID NO: 4; a heavy chain CDR2 as set forth in SEQ ID NO: 5; and a heavy chain CDR3 as set forth in SEQ ID NO: 6, and the second anticancer agent is the drug 2, The cancer may be gastric cancer. The gastric cancer may be a cancer in which c-Met is amplified. In the present invention, the term "prevention" means any act of inhibiting or delaying the progression of a cancer disease by administering a composition according to the present invention, and "treatment" means inhibiting the development of a cancer disease, alleviating or eliminating a cancer disease. In the present invention, "included as an active ingredient" means included in the composition of the present invention in a therapeutically effective amount, and the term "therapeutically effective amount" indicates the amount of the combination or pharmaceutical composition effective for treating or preventing cancer. Specifically, a "therapeutically effective amount" means an amount sufficient to treat a disease at a reasonable benefit/risk ratio applicable to medical treatment, and the effective dosage level can be determined according to factors including the type and severity of the individual, age, sex, type of disease, activity of the drug, sensitivity to the drug, administration time, administration route and excretion rate, treatment period, concurrently used drugs, and other factors well known in the medical field. The combination or pharmaceutical composition of 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 when combined. And it may be administered singly or in multiple doses. It is important to administer an amount that can achieve the maximum effect with the minimum amount without side effects by considering all of the above factors, and the administration dosage can be easily determined by those skilled in the art according to various factors such as the patient's condition, age, sex, and complications. The pharmaceutical composition of the present invention comprises an antibody or an antigen-binding fragment thereof that specifically binds to c-Met, at least one second anticancer agent among the above drugs 1 and 2, and Additionally, one or more pharmaceutically acceptable carriers may be included. Pharmaceutically acceptable carriers are those commonly used in the art, and specifically, may be, but are not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, glycine, starch, gum tragacanth, gum acacia, calcium phosphate, calcium chloride, sodium chloride, alginic acid, sodium alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidine, polyethylene glycol, cellulose, water, ethanol, syrup, methylcellulose, methyl hydroxybenzonate, propyl hydroxybenzonate, talc, magnesium stearate, magnesium aluminum silicate, silica, orange essence, strawberry essence, vanilla essence, or mineral oil. The pharmaceutical composition of the present invention may further include, in addition to the above components, a binder, an isotonic agent, an adsorbent, a disintegrant, an antioxidant, a lubricant, a filler, a solubilizer, a solubilizing agent, a lubricant, a wetting agent, a sweetener, a flavoring agent, an emulsifier, a suspending agent, a preservative, a dispersing agent, or a stabilizer. In addition, the pharmaceutical composition of the present invention may be formulated in the form of oral dosage forms such as tablets, pills, powders, granules, capsules, suspensions, emulsions, oral solutions, emulsions, syrups, external preparations, suppositories, or sterile injections using pharmaceutically acceptable carriers and excipients, and may be manufactured in unit dose form or may be manufactured by inserting into a multi-dose container. The preparation may be manufactured by a conventional method used in formulation in the art or by a method disclosed in Remington's Pharmaceutical Science ( ^19th ed., 1995), and may be formulated into various preparations depending on each disease or ingredient. The pharmaceutical composition of the present invention can be administered orally or parenterally depending on the intended method. Preferably, it can be administered orally, but is not limited thereto. Individual components included in the pharmaceutical composition of the present invention can be administered through the same route or through different routes. Non-limiting examples of dosage forms for oral administration include, but are not limited to, tablets, pills, powders, capsules, syrups, or emulsions. When the pharmaceutical composition of the present invention is in the form of oral administration, pharmaceutically acceptable carriers may include, but are not limited to, cellulose, calcium silicate, corn starch, lactose, sucrose, dextrose, calcium phosphate, stearic acid, magnesium stearate, calcium stearate, gelatin, talc, and the like, but are not limited thereto. Non-limiting examples of dosage forms for parenteral administration include, but are not limited to, injections. When the pharmaceutical composition of the present invention is in the form of a parenteral administration, water, saline, glucose aqueous solution, pseudo-saccharide aqueous solution, alcohol, glycol, ether, oil, fatty acid, fatty acid ester, glyceride, etc. can be used as a pharmaceutically acceptable carrier, but is not limited thereto. The daily dosage of the antibody or antigen-binding fragment thereof that specifically binds to c-Met of the present invention, and at least one second anticancer agent among Drug 1 and Drug 2 may be about 0.1 to 2,000 (mg/day) for an adult weighing 70 kg, but is not limited thereto, and may be administered once a day or in divided doses several times. The dosage may vary depending on the patient's health condition, age, weight, sex, dosage form, and disease severity. The pharmaceutically effective amount and effective dosage of the pharmaceutical composition of the present invention are pharmaceutically acceptable. The dosage may vary depending on the formulation method of the composition, the administration method, the administration time and/or the administration route, and may vary depending on various factors including the type and degree of the response to be achieved by the administration of the pharmaceutical composition, the type, age, weight, general health condition, symptoms or degree of the disease, sex, diet, excretion, drugs used simultaneously or simultaneously in the subject, other components of the composition, and similar factors well known in the pharmaceutical field, and a person having ordinary skill in the art can easily determine and prescribe an effective dosage for the desired treatment. The pharmaceutical composition of the present invention may be administered once a day or divided into several times. The pharmaceutical composition of the present invention may be administered as an individual therapeutic agent or additionally administered in combination with other therapeutic agents, and may be administered sequentially or simultaneously with conventional therapeutic agents. Taking all of the above factors into consideration, the amount that can obtain the maximum effect with the minimum amount without side effects can be administered, and this can be easily determined by a person having ordinary skill in the art to which the present invention pertains. The pharmaceutical composition of the present invention can exhibit excellent effects even when used alone, but can be used in combination with various methods such as drug therapy, immunotherapy, and radiation therapy to increase treatment efficiency. In the method for preventing or treating cancer of the present invention, "administration" means introducing a predetermined substance to an individual by an appropriate method. In the present invention, "individual" means all animals such as rats, mice, and livestock, including humans, that have developed or may develop cancer, and specifically may be mammals including humans, but is not limited thereto. The method for preventing or treating cancer of the present invention may be a method for preventing or treating cancer comprising administering a therapeutically effective amount of an antibody or an antigen-binding fragment thereof that specifically binds to c-Met, and at least one second anticancer agent among Drug 1 and Drug 2. The method for preventing or treating cancer of the present invention comprises not only treating the disease itself before the onset of symptoms, but also inhibiting or avoiding the symptoms thereof by administering a pharmaceutical composition comprising an antibody or an antigen-binding fragment thereof that specifically binds to c-Met, and at least one second anticancer agent among Drug 1 and Drug 2. In the management of a disease, the prophylactic or therapeutic dosage of a particular active ingredient will vary depending on the nature and severity of the disease or condition, and the route by which the active ingredient is administered. The dosage and frequency of dosage will vary depending on the age, weight, and response of the individual patient. An appropriate dosage regimen can be readily selected by one skilled in the art, taking these factors into account. In addition, the method for preventing or treating cancer of the present invention may further include administering a therapeutically effective amount of an additional active agent that helps in preventing or treating the disease together with the antibody or antigen-binding fragment thereof that specifically binds to c-Met, and at least one second anticancer agent among Drug 1 and Drug 2. The additional active agent may exhibit a synergistic or additive effect together with the antibody or antigen-binding fragment thereof that specifically binds to c-Met, and at least one second anticancer agent among Drug 1 and Drug 2. For the preparation of a pharmaceutical agent, the antibody or antigen-binding fragment thereof that specifically binds to c-Met of the present invention, and at least one second anticancer agent among Drug 1 and Drug 2 may be mixed with a pharmaceutically acceptable adjuvant, diluent, carrier, etc., and may be prepared as a composite preparation together with other active agents to exhibit a synergistic effect. The embodiments of the present invention can be modified in various different forms, and the scope of the present invention is not limited to the embodiments described below. In addition, the embodiments of the present invention are provided so that a person having average knowledge in the art can more completely explain the present invention. Furthermore, the term "including" a certain component throughout the specification does not exclude other components unless specifically stated to the contrary, but rather means that other components can be further included.

[Form for implementing the invention] Hereinafter, the present invention will be described in more detail through examples. These examples are only for explaining the present invention in more detail, and it will be apparent to those skilled in the art that the scope of the present invention is not limited by these examples according to the gist of the present invention. Example 1. Production of hybridoma cells producing c-Met specific antibodies and confirmation of their tumor cell proliferation inhibitory activity.

(1) Production and selection of hybridoma cell lines producing monoclonal antibodies against c-Met protein Immunization of immunized mice required for development of hybridoma cell lines To obtain this, human c-Met Sema domain/Fc fusion protein (produced in-house) was injected intraperitoneally into mice as an antigen. To select hybridoma cells that specifically react only with c-Met protein among the hybridoma cell population, screening was performed using an ELISA analysis method using human c-Met/His fusion protein as an antigen.

(2) The light chain and heavy chain CDR amino acid sequences of mouse antibodies obtained from the c-Met antibody-selected hybridoma cell line are shown in Tables 1 and 2, respectively.

【Table 11 Hybridoma light chain CDR

【Table 2 Hybridoma Heavy Chain CDR

(3) In vitro tumor cell proliferation inhibition activity of hybridoma c-Met antibody A c-Met specific mouse antibody obtained using a hybridoma cell line, and a chimeric antibody produced by fusing the antibody with human heavy and light chain constant regions Cell proliferation inhibition activity was tested in MKN45, a human gastric cancer cell line. Specifically, MKN45 (#JCRB0254) cell line was diluted in RPMI-1640 medium (Gibco, #A10491) containing 10% (v/v) FBS, dispensed at 2.5 x ¹⁰³ cells per well of a 96-well plate, and cultured overnight at 37 °C, 5% _CO2 . After this, the medium in each well of the plate was replaced with 100 ml of RPMI-1640 medium containing 1% (v/v) FBS, and the test antibody was serially diluted 1/10 (i.e., 100 nM, 10 nM, 1 nM, 100 pM, 10 pM, and 1 pM) from 100 nM to 1 pM at a final concentration of 100 M and added to each well. Then, the plate was cultured for 5 days under conditions of 37 °C and 5% CO ₂ , and the medium was removed, and 200 M of TCA (Trichloroacetic acid; Sigma, #T0699) solution was added to each well to fix the cells. The fixed cells were stained for 25 minutes by adding 80 M of 0.4% SRB (sulforhodamine B) solution to each well, and then washed five times with 1% acetic acid solution. Next, 150 M of 10 mM Tris solution was added to each well of the dried plate to dissolve the SRB dye, and the absorbance was measured at a wavelength of 540 nm using a microplate reader. The results for the MKN45 cell line are shown in Table 3.

상기 표 3에서 확인되는 바와 같이, 본 발명의 항 c-Met 항체인 8C4 항체, 그리고 이의 키메라 항체는 모두 우수한 종양 세포 증식 저해 활성을 갖는다. 따라서, 본 발명의 8C4 항체, 및 이의 키메라 항체, 인간화 항체, 항원에 대한 친화도를 최적화시킨 항체 등 변이체들은 암의 예방 또는 치료에 매우 유용하게 사용될 수 있다. 본 발명의 8C4 항체의 경쇄 및 중쇄 가변 영역에 대한 구체적인 컨센서스 서열을 하기 표 4에 나타내었다. 【Table 3】 Results of in vitro tumor cell proliferation inhibition activity test of hybridoma c-Met antibody

As confirmed in Table 3 above, the 8C4 antibody, an anti-c-Met antibody of the present invention, and its chimeric antibodies, all exhibit excellent tumor cell proliferation inhibition activity. Therefore, the 8C4 antibody of the present invention, as well as variants thereof, such as chimeric antibodies, humanized antibodies, and antibodies with optimized affinity for the antigen, can be very useful for the prevention or treatment of cancer. The specific consensus sequences for the light and heavy chain variable regions of the 8C4 antibody of the present invention are shown in Table 4 below.

Table 4

실시예 2. 8C4항체의 인간화 항체 제조 Consensus sequences for the light and heavy chain variable regions of the 8C4 antibody

Example 2. Production of humanized antibody 8C4

For humanization design of the 8C4 antibody heavy chain, first, Ig Blast

(http://www.ncbi.nlm.nih.gov/igblast/) were used to analyze human germline genes that are highly homologous to the heavy chain variable region gene of mouse antibody 8C4. As a result, it was confirmed that IGHV3-23 has 48% homology with 8C4 antibody at the amino acid level, and IGHV3-11 has 46% homology with 8C4 antibody at the amino acid level. CDR-H1, CDR-H2, CDR— H3 of white mouse antibody 8C4 were defined by Rabat numbering, and hu8C4-1 was produced by designing the CDR portion of mouse antibody 8C4 to be introduced into the framework of IGHV3-23. At this time, 48 (V— I), 49 (S— G), 71 (R— A),

Amino acids 73 (N—K), 78 (L—A), and 94 (K—G) are the same as those of the original mouse 8C4 antibody. The heavy chain of hu8C4-1 was finally constructed by back-mutating the amino acid sequence. In the case of hu8C4-2, the CDR portion of the mouse antibody 8C4 was designed to be introduced into the framework of IGHV3-11, and positions 48 (V—I), 49 (S—G), 71 (R—A), 73 (N—K),

Amino acids 78 (L—A) and 94 (R—G) were back-mutated to the amino acid sequence of the original mouse 8C4 antibody, ultimately constructing the heavy chain of hu8C4-2.

실시예 3. 힌지 （hinge） 변이체 제작 다음으로, 인간 IgGl 중쇄 불변 영 역의 힌지 （hinge） 서열 변이체를 제조하였다. 먼저 , 인간 IgGl 중쇄 불변 영 역의 힌지는 ' EPKSCDKTHTCPPCP （서열번호 19） '의 아미노산 서열을 가지며 , 이를 치환하여 서열번호 20 내지 서열번호 26의 아미노산 서열을 갖는 힌지 영 역 변이체를 얻었다 （표 6）. 이를 상기 실시 예 2에서 제작한 hu8C4-l , hu8C4-2 인간화 항체의 중쇄 가변영 역을 포함하는 벡터에 각각 클로닝 하였다. 【표 6] 힌지 영역 변이체 서열

힌지 서열의 차이에 따라 hu8C4 항체의 종양 세포 증식 저해 활성이 다소 차이가 있으나, 대부분 종양 세포의 증식을 효과적으로 억제하는 것을 확인하였다. 이에, 이하에서는 대표적으로 hu8C4- 1에 서열번호 20의 힌지 영역을 가지는 IgGl 인간화 항체를 hu8C4로 명명하고, 이에 대한 친화도 최적화 항체를 제작하여 그 효과를 확인하고자 하였다. 실시예 4. hu8C4의 친화도최적화항체 제작 hu8C4의 친화도 최적화 항체를 제작하기 위해, 먼저 scFv오｝ pill가 결합된 형태로 디스플레이 되는 파지미드 벡터를 사용하여 파지-디스플레이된 scFv 라이브러리를 제작하였으며, 상기 벡터의 대략적인 구조를 도 1에 도시하였다. 상기 파지미드 벡터는 IPTG-유도성 lac 프로모터의 제어 하의 항체의 scFv 절편을 포함하며 , 링커 서열은 GGGGS GGGGS GGGGS (서열번호 35)를사용하였다. 다음으로, hu8C4의 중쇄와 경쇄 CDR 부위에 다양성을 도입하기 위해서 NNK 코돈을 갖는 돌연변이 유발 올리고뉴클레오티드를 사용하였다. 이에 따라 His, HA 및 pl II가 융합된 hu8C4 scFv 라이브러리를 제작하였고, 제작된 항체 라이브러리로부터 인간 c- Met에 특이적인 항체를 선별하였다. 구체적으로, 친화도가 향상된 항체를 선별하기 위하여 경쟁적인 선별방법을 사용하였다. Dynabeads® M-280 (Thermo Fisher Scientific, 11205D)에 인간 c-Met 항원을 제조사 지침에 따라 결합시켜주었다. 항원이 결합된 비드를 수퍼블록 TBS (Superb lock Tris buffered saline, Pierce)로 2 시간 동안 차단하였다. 또한 37 T에서 재조합 파지를 밤새 성장시킨 후 원심분리하여 상등액의 파지를수퍼블록 TBS, 0.05 % Tween 20으로 2 시간동안차단하였다. 이후, 비드를 0.05 %트윈 20이 포함된 PBS로 세척하였다. 차단된 파지 용액을 세척된 비드에 첨가하고, 파지 결합을 위해 로테이터에서 2 시간 인큐베이션한 후, 비드를 0.05 % 트윈 20이 포함된 PBS로 세척하였다. 그 다음, 인간 c-Met 항원을 0.05 % 트윈 20이 포함된 PBS 1 ml에 첨가하여 로테이터에서 24 시간 인큐베이션 하였다 (Rouet R et al. (2012) Nat Protoc. 7： 364-373). 그 다음, 비드에 결합된 파지를 100 mM 트리에탄올아민으로 5 분 동안 용리하고, 용리액을 0.5 M Tris/Cl (pH 7.2)로 중화하였다. 용리된 파지 중화액을 대장균 TG1에 감염시켰다. 상기 실험을 통하여 선택된 개별 클론을 카베니실린 및 암피실린이 첨가된 2xYT 브로쓰 200 M에서 96 -웰 포맷에서 성장시키고, 배양 상층액을 ELISA에 직접 사용하여, 표적 단백질로 코팅된 플레이트에 결합하는 파지-디스플레이된 scFv를 선별하였다. 검출된 항체의 경쇄 및 중쇄 CDR 영역 아미노산 서열을 표 7 및 표 8에 나타내었으며, 대표적인 경쇄 및 중쇄 친화도 최적화 항체 가변영역 아미노산 서열을 표 9에 나타내었다. For the 8C4 antibody light chain, for humanization design, human germline genes with high homology to the light chain variable region gene of mouse antibody 8C4 were analyzed using Ig Blast (http://www.ncbi.nlm.nih.gov/igblast/). As a result, it was confirmed that IGKV1-27 had 65.3% homology with the 8C4 antibody at the amino acid level, and IGKV1-33 had 64.2% homology with the 8C4 antibody at the amino acid level. CDR-L1, CDR-L2, and CDR— L3 of the white mouse antibody 8C4 were defined by Rabat numbering, and the CDR portion of the mouse antibody 8C4 was designed to be introduced into the framework of IGKV1-33 to produce hu8C4-1, and was designed to be introduced into the framework of IGKV1-27 to produce hu8C4-2. At this time, both hu8C4-1 and hu8C4-2 had amino acid 69 (T—R) back-mutated to the amino acid sequence of the original mouse 8C4 antibody. The above 8C4 humanized antibody was expressed in 293T cells using the pCLS05 vector (Korean Patent No. 10-1420274). Specific consensus sequences for the light and heavy chain variable regions of the above hu8C4-l and hu8C4-2 humanized antibodies are shown in Table 5. 【Table 5】 Consensus sequences for the light and heavy chain variable regions of hu8C4-l and hu8C4-2 humanized antibodies.

Example 3. Production of hinge variants Next, hinge sequence variants of the human IgG1 heavy chain constant region were produced. First, the hinge of the human IgG1 heavy chain constant region has the amino acid sequence of 'EPKSCDKTHTCPPCP (SEQ ID NO: 19)', and by replacing this, hinge region variants having amino acid sequences of SEQ ID NOs: 20 to 26 were obtained (Table 6). These were each cloned into vectors containing the heavy chain variable regions of the hu8C4-1 and hu8C4-2 humanized antibodies produced in Example 2. 【Table 6】 Hinge region variant sequences

Although the tumor cell proliferation inhibitory activity of the hu8C4 antibody differed somewhat depending on the difference in the hinge sequence, it was confirmed that most tumor cell proliferation was effectively inhibited. Therefore, hereinafter, an IgG1 humanized antibody having the hinge region of SEQ ID NO: 20 in hu8C4-1 was named hu8C4, and an affinity-optimized antibody was produced to confirm its effect. Example 4. Production of hu8C4 affinity-optimized antibody In order to produce an affinity-optimized antibody of hu8C4, a phagemid vector in which scFv is displayed in a bound form was first used to produce a phage-displayed scFv library, and the general structure of the vector is shown in Fig. 1. The phagemid vector contained the scFv fragment of the antibody under the control of the IPTG-inducible lac promoter, and the linker sequence was GGGGS GGGGS GGGGS (SEQ ID NO: 35). Next, a mutagenic oligonucleotide with the NNK codon was used to introduce diversity into the heavy and light chain CDR regions of hu8C4. Accordingly, a hu8C4 scFv library fused with His, HA, and pl II was constructed, and antibodies specific for human c-Met were selected from the constructed antibody library. Specifically, a competitive selection method was used to select antibodies with improved affinity. Human c-Met antigen was bound to Dynabeads® M-280 (Thermo Fisher Scientific, 11205D) according to the manufacturer's instructions. The antigen-bound beads were blocked with Superblock Tris-buffered saline (TBS, Pierce) for 2 hours. In addition, recombinant phages were grown overnight at 37 T, centrifuged, and the supernatant phages were blocked with Superblock TBS, 0.05% Tween 20 for 2 hours. The beads were then washed with PBS containing 0.05% Tween 20. The blocked phage solution was added to the washed beads, and incubated on a rotator for 2 hours for phage binding, and then the beads were washed with PBS containing 0.05% Tween 20. Then, human c-Met antigen was added in 1 ml of PBS containing 0.05% Tween 20, and incubated on a rotator for 24 hours (Rouet R et al. (2012) Nat Protoc. 7: 364-373). Next, the phages bound to the beads were eluted with 100 mM triethanolamine for 5 min, and the eluate was neutralized with 0.5 M Tris/Cl (pH 7.2). The eluted phage neutralization solution was used to infect E. coli TG1. Individual clones selected through the above experiments were grown in 96-well format in 2xYT broth 200 M supplemented with carbenicillin and ampicillin, and the culture supernatant was directly used in ELISA to select phage-displayed scFvs that bound to plates coated with the target protein. The light and heavy chain CDR region amino acid sequences of the detected antibodies are shown in Tables 7 and 8, and representative light and heavy chain affinity-optimized antibody variable region amino acid sequences are shown in Table 9.

【표 8]

Table 7

Table 8

본 발명의 친화도 최적화항체는 모체가 되는 hu8C4 항체의 친화도를 최적화한 것이고 , 이들은 모두 선별과정에서 항원 친화도를 기반으로 선별된 것인바, 제시된 친화도 최적화 항체 , 그리고 제시된 중쇄 및 경쇄 가변영 역 CDR을 조합한 항체들은 모두 동등한 효과가 있을 것으로 예상된다. 추가적인 실험을 위해 상기 경쇄와 중쇄 가변영 역을 조합하여 10 종의 친화도 최적화 항체를 제작하였다. 구체적인 경쇄 및 중쇄 서열 조합을 표 10에 나타내었다. 【Table 2: List of variable region sequences of light and heavy chain affinity-optimized antibodies

The affinity-optimized antibodies of the present invention are those that have optimized the affinity of the parent hu8C4 antibody, and all of these were selected based on antigen affinity during the selection process. Therefore, the proposed affinity-optimized antibodies and antibodies combining the proposed heavy and light chain variable region CDRs are all expected to have equivalent effects. For further experiments, 10 types of affinity-optimized antibodies were produced by combining the light and heavy chain variable regions. Specific light and heavy chain sequence combinations are shown in Table 10.

실시예 5. 이중특이적 항체 제작 c-Met 및 EGFR에 특이적으로 결합하는 이중항체를 제작하기 위하여, EGFR에 특이적으로 결합하는 것으로 알려진 Erbitux （세툭시맙）와 Vect ibix （파니투무맙） scFv 절편을 각각 본원발명의 c-Met 항체의 중쇄 C-말단에 GGGGSGGGGS （서열번호 294） 커넥터로 연결하여 이중항체를 제작하였다. 상기 유內의 안정성을 증가시키기 위하여 중쇄 44 번째 잔기와 경쇄 100 번째 잔기를 시스틴으로치환하였다 （Reiter Y. et al., Biochemistry 33（18）:5451- 5459 （1994）） . Erbitux와 Vect ibix scFv 서열, 이중항체 중쇄 아미노산 서열 및 이중항체 가변영역 조합을 아래 표 11 및 표 12에 나타내었다. 【Table 10】List of variable region combination sequences for affinity-optimized antibodies

Example 5. Production of bispecific antibodies In order to produce a bispecific antibody that specifically binds to c-Met and EGFR, scFv fragments of Erbitux (cetuximab) and Vect ibix (panitumumab), known to specifically bind to EGFR, were linked to the heavy chain C-terminus of the c-Met antibody of the present invention using a GGGGSGGGGS (SEQ ID NO: 294) connector to produce a bispecific antibody. In order to increase the stability of the above-mentioned molecules, the 44th residue of the heavy chain and the 100th residue of the light chain were substituted with cysteine (Reiter Y. et al., Biochemistry 33 (18): 5451- 5459 (1994)). The Erbitux and Vect ibix scFv sequences, bispecific antibody heavy chain amino acid sequences, and bispecific antibody variable region combinations are shown in Tables 11 and 12 below.

Table 111

그 다음, Erbitux오｝ Vect ibix scFv절편을 연결한 이중항체의 in vitro 항암 효능을 U-87 MG, NCI-H1993, NCI-H292 및 NCI-H820 종양 세포주에서 평가하였다. 구체적으로 각 세포주를 10 % （v/v） FBS를 함유하는 RPMI-1640 배지 （Gibco, #A10491）에 희석하여 96 -웰 플레이트의 각 웰에 2.0 x 10³ 개 씩 분주해 넣은 후【Table 12】List of dual antibody variable region combination sequences

Next, the in vitro anticancer efficacy of the bispecific antibody linked to the Erbitux® Vect ibix scFv fragment was evaluated in U-87 MG, NCI-H1993, NCI-H292, and NCI-H820 tumor cell lines. Specifically, each cell line was diluted in RPMI-1640 medium (Gibco, #A10491) containing 10% (v/v) FBS and dispensed at 2.0 x ¹⁰³ cells per well of a 96-well plate.

The cells were cultured overnight at 37 °C, 5% CO ₂ conditions. Afterwards, each well of the plate was replaced with 100 M serum-free medium and cultured for 18 hours at 37 °C, 5% CO ₂ conditions. Then, the medium was replaced with 100 M RPMI-1640 medium containing 2% (v/v) FBS or HGF 50 ng/ml. After replacement, the test antibody was serially diluted 1/10 (i.e., 100 nM, 10 nM, 1 nM, 100 pM, 10 pM, and 1 pM) from 100 nM to 0.001 nM at a final concentration, and 100 M was added to each well. Then, the plate was cultured for 5 days under conditions of 37 °C and 5% CO ₂ , and the medium was removed, and 200 M TCA solution was added to each well to fix the cells. The cells on the plate were stained according to the conventional SRB colorimetric assay method, and the absorbance of each well was measured at a wavelength of 540 nm using a microplate reader. The results of proliferation inhibition activity in each cell line are shown in Tables 13 and 14.

【Table 13】 In vitro cancer cell line proliferation inhibition activity by dual antibodies

종양 세포 증식 저해 활성 평가 결과 , 제작된 이중항체간의 효능 차이는 없음을 확인하였다. 대표적인 예시로, 상기 hu8C4 x Vect ibix scFv 항체를 CM01로 명명하고 , 이후의 실험을 진행하였다. 실시예 6. 에피토프 맵핑 본 발명의 이중항체 CM01의 인간 c-Met 항원에 대한 에피토프를 규명하기 위하여 오송첨단의료산업진흥재단 분자모델설계지원팀 (Korea)에 분석을 의뢰하였다 . 수소-중수소 교환 질량 분광법 (Hydrogen-deuter ium exchange Mass Spectrometry, HDX- MS)을사용하여 분석을수행하였다. c-Met 세마도메인은 a/p 두 사슬로 구성되어 있어서 두 사슬에 대한 각각의 커버리지 (coverage)를 확인하였으며, 시료 내 이황화결합이 다수 존재하는 이유로 퀀치 홀딩 (quench holding) 시간, TCEP 농도, 펩신 농도 등을 조절하여 펩타이드 커버리지를 최적화하였다. 최종적으로 퀀치 버퍼 (Quench buffer) 조건은 100 mM K. Phosphate, 125 mM TCEP, 0.5 M Guanidine-HCl , pH 2.66으로 실험을 수행하였다. 항원과 항체는 각각 3.3 mg/ml과 65 mg/ml 농도로준비하였고, 37 pmol cMET 항원과 36 pmol 항체를 실험 전에 3 시간 이상 미리 결합시켰다. 중수소 라벨링 버퍼 (deuterium 1 abe 1 i ng buffer)를 0 분, 0.33 분, 10 분, 60 분, 240 분 동안 반응시켰다. 각각의 라벨링 시간에 맞춰 퀀치 버파로 라벨링을 중단하고 vortexing한 후 액체질소에 바로 얼려 분석 전까지 -80 T에 보관하였다. 질량분석기 (MS)로 펩신 컬럼에 로딩하여 분석하였다. 분석 결과, 본 발명의 이중항체 CM01은 인간 c-Met 세마도메인 p 사슬의 Y321-L329 (서열번호 313), 1333〜 1341 (서열번호 314), P366-D372 (서열번호 315), 및 Q464〜 S474 (서열번호 316) 4 개의 부위에 입체적형태의 에피토프에 결합함을 확인하였다 (표 15). PyMOL프로그램을 이용하여 인간 c-Met 항원 (PDB No. 4K3J) 3 차 구조상에 표지하여 도 2에 나타내었다. Table 14 In vitro cancer cell line proliferation inhibition activity by dual antibodies

As a result of the tumor cell proliferation inhibition activity evaluation, it was confirmed that there was no difference in efficacy between the produced dual antibodies. As a representative example, the hu8C4 x Vect ibix scFv antibody was named CM01, and the subsequent experiments were conducted. Example 6. Epitope Mapping To identify the epitope for the human c-Met antigen of the dual antibody CM01 of the present invention For this purpose, the analysis was requested to the Molecular Model Design Support Team of the Osong Medical Innovation Foundation (Korea). The analysis was performed using hydrogen-deuter ium exchange mass spectrometry (HDX-MS). Since the c-Met semadomain is composed of two chains, a/p, the coverage for each chain was confirmed, and because there were many disulfide bonds in the sample, the peptide coverage was optimized by adjusting the quench holding time, TCEP concentration, and pepsin concentration. The final quench buffer condition was 100 mM K. Phosphate, 125 mM TCEP, 0.5 M Guanidine-HCl, pH 2.66. Antigen and antibody were prepared at concentrations of 3.3 mg/ml and 65 mg/ml, respectively, and 37 pmol cMET antigen and 36 pmol antibody were pre-bound for more than 3 hours before the experiment. Deuterium labeling buffer (deuterium 1 abe 1 i ng buffer) was reacted for 0, 0.33, 10, 60, and 240 minutes. Labeling was stopped with quench buffer at each labeling time, vortexed, and immediately frozen in liquid nitrogen and stored at -80 T until analysis. The samples were loaded onto a pepsin column and analyzed by mass spectrometry (MS). As a result of the analysis, it was confirmed that the bispecific antibody CM01 of the present invention binds to the epitopes in the three-dimensional form at four sites of Y321-L329 (SEQ ID NO: 313), 1333-1341 (SEQ ID NO: 314), P366-D372 (SEQ ID NO: 315), and Q464-S474 (SEQ ID NO: 316) of the human c-Met semadomain p chain (Table 15). Using the PyMOL program, human c-Met antigen (PDB No. 4K3J) 3 The structure of the car is indicated in Fig. 2.

실시예 7. 위암에서 항- c- Met 항체의 병용 【Table 15】 Amino acid sequence of the epitope region

Example 7. Combination of anti-c-Met antibodies in gastric cancer

(1) In vitro combination test To confirm the tumor cell proliferation inhibition activity by the combined effect of CM01 targeting c-Met and SN-38, a topoisomerase inhibitor, gastric cancer cell lines Hs746T (ATCC, #HTB-135), SNU-719 (Korea Cell Line Bank, #719), and MKN45 (#JCRB-0254) were used. Hs746T has c-Met exl4 skipping mutation and gene amplification, MKN45 has c-Met gene amplification, and SNU-719 has wild-type EGFR and c-Met genes. Each cell line was diluted in culture medium containing 10% (v/v) FBS, dispensed into each well of a 96-well plate at a density of 3 x ¹⁰³ , and cultured overnight at 37 °C, 5% _CO2 . Afterwards, CM01 and SN-38 were added to each well in culture medium containing 2% (v/v) FBS. Solutions containing them alone or in combination were made, diluted using culture medium, and serially diluted by 1/3, and added to each well at 100 M each. Then, the plate was cultured for 5 days under conditions of 37 °C and 5% CO ₂ , and the medium was removed, and 100 M of TCA solution was added to each well to fix the cells. After that, the cells on the plate were stained according to the conventional SRB colorimetric assay method, and the absorbance of each well was measured at a wavelength of 540 nm using a microplate reader. The degree of cell proliferation was quantified by calculating the difference in absorbance, and the combination index (CI) was calculated using Compusyn software (Table 16; Leukemia&Lymphoma, 2008, 49(11): 2059).

시험 결과를 하기 표 17 내지 표 19 및 도 3 내지 도 5에 나타내었다. 【표 17]

Table 16

The test results are shown in Tables 17 to 19 and Figures 3 to 5. Table 17

Table 18

시험 결과 , Hs746T 세포주에서 CM01과 SN-38 단독 처리 시의 IC50은 0.24 nM과 9.67 nM 이었으며 , 병용시의 IC50은 0.15 nM로 측정되었다. 두 약물의 시너지 효과를 정량적으로 측정하기 위해서 CI 값을 확인한 결과 0.103- 0.252로 강한 시너지 효과가 확인되었다. 다음으로 SNU-719 세포주에서 CM01의 IC50은 100 nM 초과 , SN- 38의 IC50은 4.34 nM 이었으며 , 병용 시에 IC50은 0.59 nM로 측정되었다. CI 값은 0.136- 0.153으로 측정되어 강한 시너지 효과가 있는 것으로 확인되었다. 마지막으로, MNK-45 세포주에서 CM01과 SN- 38의 병용 처리는 각 약물의 단독 처리보다 종양 세포 증식 저해능이 우수한 것으로 농도 의존적으로 확인되었다. CM01과 SN-38 단독 처리군의 IC₅₀ 값은 각각 0.373 nM과 0.780 nM 이었으며 , 병용 처리군의 IC50값은 0.019 nM로 병용 처리군의 우수한 저해 효과를 확인하였다. CI 값은 약 0.1로 강한 세포 증식 저해 시너지 효과가 확인되었다. Table 19

As a result of the test, the IC50 of CM01 and SN-38 alone in the Hs746T cell line was 0.24 nM and 9.67 nM, and the IC50 of combined use was measured to be 0.15 nM. Synergy of the two drugs To quantitatively measure the effect, the CI value was checked and a strong synergistic effect was confirmed with 0.103-0.252. Next, in the SNU-719 cell line, the IC50 of CM01 was more than 100 nM, the IC50 of SN-38 was 4.34 nM, and when used in combination, the IC50 was measured to be 0.59 nM. The CI value was measured to be 0.136-0.153, confirming a strong synergistic effect. Lastly, in the MNK-45 cell line, the combined treatment of CM01 and SN-38 was confirmed to have a superior tumor cell proliferation inhibition effect than the treatment of each drug alone in a concentration-dependent manner. The IC ₅₀ values of the CM01 and SN-38 single treatment groups were 0.373 nM and 0.780 nM, respectively, and the IC 50 value of the combination treatment group was 0.019 nM, confirming the excellent inhibitory effect of the combination treatment group. The CI value was approximately 0.1, confirming a strong synergistic effect of inhibiting cell proliferation.

(2) In vivo combination test

MKN-45 cells were cultured in RPMI1640 medium supplemented with 10% (v/v) FBS in a cell culture flask and cultured in a 37 °C, 5% CO ₂ incubator. The cultured cells were detached, centrifuged (1,000 rpm, 5 min), and the supernatant was removed. The prepared cells were resuspended in serum-free medium and prepared at a concentration of 5.0 x 10 ⁷ cells/mL. They were subcutaneously inoculated into BALB/c nude mice at a concentration of 1.0 x 10 ⁷ cells/0.2 mL per mouse. When the tumor volume reached approximately 150 mF, each group was randomly distributed to ensure uniform tumor size. Drug administration was performed by intraperitoneal injection of 15 mg/kg irinotecan as a topoisomerase inhibitor and 10 mg/kg CM twice a week. Tumor volume was measured three times a week from the day of group division, and the long and short axes of the tumors were measured using a caliper. It was measured. Tumor volume (mm ³ ) and tumor growth inhibition rate (%) were calculated using the following formula, and the tumor volume compared to the control group was statistically analyzed using Dunnett's T3 multiple comparison test. Tumor volume (mm ³ ) = ab ² /2 (a: long axis length, b: short axis length), Tumor growth inhibition rate (%) = {1 - (average tumor volume of drug administration group / average tumor volume of control group)} X 100. The test results are shown in Table 20 and Figure 6 below.

Table 20

— MTV： Mean tumor volume

— TGI (%): tumor growth inhibition As confirmed in Table 20 and Figure 6, it was found that when the anti-c-Met antibody of the present invention and irinotecan were administered together, a synergistic effect was shown by significantly reducing tumor growth compared to when administered alone. While specific aspects of the present invention have been described in detail above, it will be apparent to those skilled in the art that such specific descriptions are merely preferred embodiments and that the scope of the present invention is not limited thereby. Accordingly, the actual scope of the present invention will be defined by the appended claims and their equivalents.

Claims

86 【Scope of claims】 【Claim 11】 (A) an antibody or an antigen-binding fragment thereof that specifically binds to hepatocyte growth factor receptor (c-Met) that binds to at least one epitope region described by an amino acid sequence selected from the group consisting of SEQ ID NO: 313, SEQ ID NO: 314, SEQ ID NO: 315, and SEQ ID NO: 316, and (B) a pharmaceutical composition for preventing or treating cancer, comprising at least one of the following (1) drug 1 and (2) drug 2 as an active ingredient: (1) drug 1: SN-38 represented by the following chemical formula 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof [Chemical Formula 1]

(2) Drug 2: Irinotecan represented by the following chemical formula 2, an optical isomer thereof, or a pharmaceutically acceptable salt thereof [Chemical Formula 2] 87

【A pharmaceutical composition according to claim 1, wherein (B) is the drug 1. 【Claim 3】 A pharmaceutical composition according to claim 2, wherein the cancer is gastric cancer. 【Claim 4】 A pharmaceutical composition according to claim 2, wherein the antibody comprises a light chain variable region comprising a light chain CDR1 as set forth in SEQ ID NO: 1; a light chain CDR2 as set forth in SEQ ID NO: 2; a light chain CDR3 as set forth in SEQ ID NO: 3; and a heavy chain variable region comprising a heavy chain CDR1 as set forth in SEQ ID NO: 4; a heavy chain CDR2 as set forth in SEQ ID NO: 5; and a heavy chain CDR3 as set forth in SEQ ID NO: 6, or an affinity-optimized antibody thereof. 88 【Claim 5】 In claim 4, the affinity-optimized antibody comprises a light chain variable region comprising a light chain CDR1 as set forth in SEQ ID NO: 1; a light chain CDR2 as set forth in SEQ ID NO: 2; a light chain CDR3 as set forth in SEQ ID NO: 3, and a heavy chain variable region comprising a heavy chain CDR1 as set forth in SEQ ID NO: 4; a heavy chain CDR2 as set forth in SEQ ID NO: 5; and a heavy chain CDR3 as set forth in SEQ ID NO: 6, wherein at least one amino acid sequence is substituted in the antibody, wherein (i) G at the 1st position of the light chain CDR1 is A, E, K, L, N, R, S, V, or W; A at the 2nd position is C, G, I, P, S, T, or V; E at the 3rd position is G, M, N, P, Q, R, S, or T; E at the 4th position is A, D, F, G, H, K, M, Q, R, S, T or V; N in the 5th position is A, D, E, G, K, L, P, Q, R, S, T or V; I in the 6th position is A, F, L, M, Q, R, S, T or V; Y in the 7th position is F, H, R or V; or G at the 8th position is replaced with D, F, H, M, N, R, S, T or V; (ii) G at the 1st position of light chain CDR2 is replaced with D, F, H, K, P, Q, S, V or Y; T at the 3rd position is replaced with Q; or N at the 4th position is replaced with G; (iii) Q at the 1st position of light chain CDR3 is replaced with E, G, I, M or N; N at the 2nd position is replaced with A, D, E, H, L, Q, S or T; V at the 3rd position is replaced with I, L, M, N, Q, S or T; L at the 4th position is replaced with F, H, I, M, R, S, V, W or Y; L at the 5th position is replaced with S is C, D, E, F, G, H, K, L, N, Q, R, T, V or Y; S in the 6th position is D, E, F, G, H, I, L, M, N, P, Q, R, T, V or Y; P in the 7th position is A, D, E, G, N, Q, S or V; Y in the 8th position is E, F, L, M or Q; or T in the 9th position is D, F, G, I, L, N, S, V, W or Y; (iv) 日 at the 1st position of heavy chain CDR1 is replaced with G or Q; Y at the 2nd position is replaced with Q; or I at the 4th position is replaced with A or Y; (v) 日 at the 3rd position of heavy chain CDR2 is replaced with G or Q; or Y at the 4th position is replaced with Q; or I at the 4th position is replaced with A or Y; F in the 89th position is D, E, W or Y; G in the 5th position is D, H or Y; E in the 6th position is F, P, W or Y; G in the 7th position is A, F, L, N or T; N in the 8th position is F, P, S, T or Y; T in the 9th position is A, D, E, F, G, H, L, P, S or V; H in the 10th position is A, D, F, M, R, S, T, V, W or Y; F in the 11th position is G, H, I , L, M, N, P, Q, V or Y; ▽} A, D, G, H, I , L, P, T or V; A in the 13th position is D, E, F, G, H, I , K, L, M, P, R, S, T, V or Y; R in the 14th position is A, E, G, H, L, N, P, Q, S, W or Y； F in the 15th position is D, E, G, L, M, P, R, S, V or W； The 16th position is A, E, F, G, H, L, R, S, T, V or Y; or the 17th position is G E, F, H, L, M, N, P, Q, R, S, T, V or W; or (vi) G at position 1 of heavy chain CDR3 is E, F, H, N, Q, V or W; 日 at position 2 is E; Y at position 3 is L, Q, T or V; G at the 4th position is W; F at the 5th position is L or Y; L at the 6th position is Q, S or Y; or Y at the 7th position is replaced with C, L, M, N or this, wherein light chain CDR1 is 0 to 5, light chain CDR2 is 0 to 1, light chain CDR3 is 0 to 7, heavy chain A pharmaceutical composition comprising 0 to 1 substitution in CDR1, 0 to 11 substitutions in heavy chain CDR2, and 0 to 6 substitutions in heavy chain CDR3. 【Claim 6】 A pharmaceutical composition according to claim 2, wherein the antibody further comprises an antibody or an antigen-binding fragment thereof that specifically binds to epidermal growth factor receptor (EGFR). 90 【Claim 7】 A pharmaceutical composition according to claim 2, wherein the antibody or antigen-binding fragment thereof comprises a light chain variable region as set forth in SEQ ID NO: 11 and any one of SEQ ID NOs: 288 to 293; and a heavy chain variable region as set forth in any one of SEQ ID NOs: 298 to 312. 【Claim 8】 A pharmaceutical composition according to claim 2, wherein the antibody or antigen-binding fragment thereof comprises a light chain variable region comprising an amino acid sequence set forth in SEQ ID NO: 11 and a heavy chain variable region comprising an amino acid sequence set forth in SEQ ID NO: 308. 【A pharmaceutical composition according to claim 2, wherein the cancer is a cancer that overexpresses c-Met, amplifies c-Met, or activates c-Met. 【Claim 10】 A pharmaceutical composition according to claim 1, wherein (B) is the drug 2. 【Claim 11】 91 A pharmaceutical composition according to claim 10, wherein the cancer is gastric cancer. 【Claim 1 is a pharmaceutical composition according to claim 10, wherein the antibody comprises a light chain variable region comprising a light chain CDR1 as set forth in SEQ ID NO: 1; a light chain CDR2 as set forth in SEQ ID NO: 2; a light chain CDR3 as set forth in SEQ ID NO: 3; and a heavy chain variable region comprising a heavy chain CDR1 as set forth in SEQ ID NO: 4; a heavy chain CDR2 as set forth in SEQ ID NO: 5; and a heavy chain CDR3 as set forth in SEQ ID NO: 6, or an affinity-optimized antibody thereof. 【Claim 13】 The antibody of claim 12, wherein the affinity-optimized antibody comprises a light chain variable region comprising a light chain CDR1 as set forth in SEQ ID NO: 1; a light chain CDR2 as set forth in SEQ ID NO: 2; a light chain CDR3 as set forth in SEQ ID NO: 3, and a heavy chain variable region comprising a heavy chain CDR1 as set forth in SEQ ID NO: 4; a heavy chain CDR2 as set forth in SEQ ID NO: 5; and a heavy chain CDR3 as set forth in SEQ ID NO: 6, wherein at least one amino acid sequence is substituted in the antibody, wherein (i) G at the 1st position of the light chain CDR1 is A, E, K, L, N, R, S, V, or W; A at the 2nd position is C, G, I, P, S, T, or V; S at the 3rd position is G, M, N, P, Q, R, S, or T; E at the 4th position is A, D, F, G, H, K, M, Q, R, S, T, or V; N at the 5th position is A, D, E, G, K, L, P, Q, R, S, T or V; I in the 6th position is A, F, L, M, Q, R, S, T or V; Y in the 7th position is F, H, R or V; or G in the 8th position is D, F, H, M, N, R, S, T or 92 Substitution with V; ( i i ) G at the 1st position of light chain CDR2 is replaced with D, F, H, K, P, Q, S, V or Y; T at the 3rd position is replaced with Q; or N at the 4th position is replaced with ; ( i i i ) Q at the 1st position of light chain CDR3 is E, G, I, M or N; 凡 at the 2nd position is A, D, E, H, L, Q, S or T; V at the 3rd position is I, L, M, N, Q, S or T; L at the 4th position is F, H, I, M, R, S, V, W or Y; S at the 5th position is C, D, E, F, G, H, K, L, N, Q, R, T, V or Y; S at the 6th position is D, E, F, G, H, I, L, M, N, P, Q, R, T, V or Y; P at the 7th position is A, D, E, G, N, Q, S or V; Y at the 8th position is replaced with E, F, L, M or Q; or T at the 9th position is replaced with D, F, G, I, L, N, S, V, W or Y; (iv) 日 at the 1st position of heavy chain CDR1 is replaced with G or Q; Y at the 2nd position is replaced with Q; or I at the 4th position is replaced with A or I; (v) The 3rd position of the heavy chain CDR2 is D, E, W, or Y; the 5th position is D, H, or Y; the 6th position is F, P, W, or Y; the G at the 7th position is A, F, L, N, or T; the 8th position is F, P, S, T, or Y; the T at the 9th position is A, D, E, F, G, H, L, P, S, or V; the H at the 10th position is A, D, F, M, R, S, T, V, W, or Y; the F at the 11th position is G, H, I, L, M, N, P, Q, V, or Y; the S at the 12th position is A, D, G, H, I, L, P, T, or V; the A at the 13th position is D, E, F, G, H, I, K, L, M, P, R, S, T, V, or Y; R in the 14th position is A, E, G, H, L, N, P, Q, S, W or Y; F in the 15th position is D, E, G, L, M, P, R, S, V or W; V in the 16th position} A, E, F, G, H, L, R, S, T, V or Y; or G in the 17th position is replaced by E, F, H, L, M, N, P, Q, R, S, T, V or W; or (vi) G at the 1st position of heavy chain CDR3 is E, F, H, N, Q, V or W; 日 at the 2nd position is E; The 3rd position's O is L, Q, T or V; the 4th position's E is W; the 5th position's F is L or 93 Y; L at the 6th position is replaced with Q, S or Y; or Y at the 7th position is replaced with C, L, M, N or this, wherein the light chain CDR1 comprises 0 to 5 substitutions, the light chain CDR2 comprises 0 to 1 substitution, the light chain CDR3 comprises 0 to 7 substitutions, the heavy chain CDR1 comprises 0 to 1 substitution, the heavy chain CDR2 comprises 0 to 11 substitutions, and the heavy chain CDR3 comprises 0 to 6 substitutions. 【Claim 14】 A pharmaceutical composition according to claim 10, wherein the antibody further comprises an antibody or an antigen-binding fragment thereof that specifically binds to epidermal growth factor receptor (EGFR). 【Claim 15】 A pharmaceutical composition according to claim 10, wherein the antibody or antigen-binding fragment thereof comprises a light chain variable region as set forth in SEQ ID NO: 11 and any one of SEQ ID NOs: 288 to 293; and a heavy chain variable region as set forth in any one of SEQ ID NOs: 298 to 312. 【Claim 16】 A pharmaceutical composition according to claim 10, wherein the antibody or antigen-binding fragment thereof comprises a light chain variable region comprising an amino acid sequence set forth in SEQ ID NO: 11 and a heavy chain variable region comprising an amino acid sequence set forth in SEQ ID NO: 308. 【Claim 17】 A pharmaceutical composition according to claim 10, wherein the cancer is a cancer that overexpresses c-Met, amplifies c-Met, or activates c-Met. 【Claim 18】 (A) A pharmaceutical composition for preventing or treating cancer, comprising as an active ingredient an antibody or an antigen-binding fragment thereof that specifically binds to hepatocyte growth factor receptor (c-Met) that binds to at least one epitope region described in an amino acid sequence selected from the group consisting of SEQ ID NO: 313, SEQ ID NO: 314, SEQ ID NO: 315, and SEQ ID NO: 316, wherein the pharmaceutical composition is administered to a subject in combination with (B) at least one of the following (1) drug 1 and (2) drug 2: (1) Drug 1: SN-38 represented by the following chemical formula 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof [Chemical Formula 1]

(2) Drug 2: Irinotecan represented by the following chemical formula 2, Optical isomer or pharmaceutically acceptable salt thereof [Chemical Formula 2]

【Claim 1: A pharmaceutical composition according to claim 18, wherein (B) is the drug 1. 【Claim 20】 A pharmaceutical composition according to claim 19, wherein the cancer is gastric cancer. 【Claim 21】 The antibody of claim 19, wherein the antibody comprises a light chain variable region comprising a light chain CDR1 as set forth in SEQ ID NO: 1; a light chain CDR2 as set forth in SEQ ID NO: 2; a light chain CDR3 as set forth in SEQ ID NO: 3, and a heavy chain variable region comprising a heavy chain CDR1 as set forth in SEQ ID NO: 4; a heavy chain CDR2 as set forth in SEQ ID NO: 5; and a heavy chain CDR3 as set forth in SEQ ID NO: 6. A pharmaceutical composition comprising 96 or an affinity-optimized antibody thereof. 【Claim 2 is an antibody according to claim 21, wherein the affinity-optimized antibody comprises a light chain variable region comprising a light chain CDR1 as set forth in SEQ ID NO: 1; a light chain CDR2 as set forth in SEQ ID NO: 2; a light chain CDR3 as set forth in SEQ ID NO: 3, and a heavy chain variable region comprising a heavy chain CDR1 as set forth in SEQ ID NO: 4; a heavy chain CDR2 as set forth in SEQ ID NO: 5; and a heavy chain CDR3 as set forth in SEQ ID NO: 6, wherein at least one amino acid sequence is substituted in the antibody, wherein (i) G at the 1st position of the light chain CDR1 is A, E, K, L, N, R, S, V, or W; A at the 2nd position is C, G, I, P, S, T, or V; S at the 3rd position is G, M, N, P, Q, R, S, or T; E at the 4th position is A, D, F, G, H, K, M, Q, R, S, T, or V; N at the 5th position is A, D, E, G, K, L, P, Q, R, S, T or V; I at the 6th position is replaced with A, F, L, M, Q, R, S, T or V; Y at the 7th position is replaced with F, H, R or V; or G at the 8th position is replaced with D, F, H, M, N, R, S, T or V; (ii) G at the 1st position of the light chain CDR2 is replaced with D, F, H, K, P, Q, S, V or Y; T at the 3rd position is replaced with Q; or N at the 4th position is replaced with ; (iii) Q at the 1st position of the light chain CDR3 is E, G, I, M or N; V at the 2nd position is A, D, E, H, L, Q, S or T; V at the 3rd position is I, L, M, N, Q, S or T; L at the 4th position is F, H, I, M, R, S, V, W or Y; S at the 5th position is C, D, E, F, G, H, K, L, N, Q, R, T, V or Y; S at the 6th position is D, E, F, G, H, I, L, M, N, P, Q, R, T, V or Y; P at the 7th position is A, D, E, G, N, Q, S or V; Y in the 8th position is E, F, L, M or Q; or Y in the 9th position is T at position 97 is replaced with D, F, G, I, L, N, S, V, W or Y; (iv) 日 at position 1 of heavy chain CDR1 is replaced with G or Q; Y at position 2 is replaced with Q; or I at position 4 is replaced with A or Y; (v) The 3rd position of the heavy chain CDR2 is D, E, W, or Y; the 5th position is D, H, or Y; the 6th position is F, P, W, or Y; the G at the 7th position is A, F, L, N, or T; the 8th position is F, P, S, T, or Y; the T at the 9th position is A, D, E, F, G, H, L, P, S, or V; the H at the 10th position is A, D, F, M, R, S, T, V, W, or Y; the F at the 11th position is G, H, I, L, M, N, P, Q, V, or Y; the S at the 12th position is A, D, G, H, I, L, P, T, or V; the A at the 13th position is D, E, F, G, H, I, K, L, M, P, R, S, T, V, or Y; R in the 14th position is replaced by A, E, G, H, L, N, P, Q, S, W or Y; F in the 15th position is replaced by D, E, G, L, M, P, R, S, V or W; V in the 16th position is replaced by A, E, F, G, H, L, R, S, T, V or Y; or G in the 17th position is replaced by E, F, H, L, M, N, P, Q, R, S, T, V or W; Or (vi) a pharmaceutical composition wherein G at the 1st position of the heavy chain CDR3 is replaced with E, F, H, N, Q, V or W; 日 at the 2nd position is replaced with E; 오 at the 3rd position is replaced with L, Q, T or V; 으 at the 4th position is replaced with W; フ at the 5th position is replaced with L or Y; ל at the 6th position is replaced with Q, S or Y; or י at the 7th position is replaced with C, L, M, N or Y, wherein the light chain CDR1 comprises 0 to 5 substitutions, the light chain CDR2 comprises 0 to 1 substitution, the light chain CDR3 comprises 0 to 7 substitutions, the heavy chain CDR1 comprises 0 to 1 substitution, the heavy chain CDR2 comprises 0 to 11 substitutions, and the heavy chain CDR3 comprises 0 to 6 substitutions. 【Claim 23】 In claim 19, the antibody further binds to epidermal growth factor receptor (EGFR). A pharmaceutical composition further comprising an antibody or antigen-binding fragment thereof that specifically binds to 98. 【Claim 24】 A pharmaceutical composition according to claim 19, wherein the antibody or antigen-binding fragment thereof comprises a light chain variable region as set forth in SEQ ID NO: 11 and any one of SEQ ID NOs: 288 to 293; and a heavy chain variable region as set forth in any one of SEQ ID NOs: 298 to 312. 【Claim 25】 A pharmaceutical composition according to claim 19, wherein the antibody or antigen-binding fragment thereof comprises a light chain variable region comprising an amino acid sequence set forth in SEQ ID NO: 11 and a heavy chain variable region comprising an amino acid sequence set forth in SEQ ID NO: 308. 【Claim 26】 A pharmaceutical composition according to claim 19, wherein the cancer is a cancer that overexpresses c-Met, amplifies c-Met, or activates c-Met. 【Claim 27】 A pharmaceutical composition according to claim 18, wherein (B) is the drug 2. 99 【Claim 28】 A pharmaceutical composition according to claim 27, wherein the cancer is gastric cancer. 【Claim 2: A pharmaceutical composition according to claim 27, wherein the antibody comprises a light chain variable region comprising a light chain CDR1 as set forth in SEQ ID NO: 1; a light chain CDR2 as set forth in SEQ ID NO: 2; a light chain CDR3 as set forth in SEQ ID NO: 3; and a heavy chain variable region comprising a heavy chain CDR1 as set forth in SEQ ID NO: 4; a heavy chain CDR2 as set forth in SEQ ID NO: 5; and a heavy chain CDR3 as set forth in SEQ ID NO: 6, or an affinity-optimized antibody thereof. 【Claim 30】 The antibody of claim 29, wherein the affinity-optimized antibody comprises a light chain variable region comprising a light chain CDR1 set forth in SEQ ID NO: 1; a light chain CDR2 set forth in SEQ ID NO: 2; a light chain CDR3 set forth in SEQ ID NO: 3, and a heavy chain variable region comprising a heavy chain CDR1 set forth in SEQ ID NO: 4; a heavy chain CDR2 set forth in SEQ ID NO: 5; and a heavy chain CDR3 set forth in SEQ ID NO: 6, wherein at least one amino acid sequence is substituted in the antibody, wherein (i) G at the 1st position of the light chain CDR1 is A, E, K, L, N, R, S, V, or W; A at the 2nd position is C, G, I, P, S, T, or V; S at the 3rd position is G, M, N, P, Q, R, S, or T; E at the 4th position is A, D, F, G, H, K, M, Q, R, S, T, or V; N at the 5th position is A, D, E, G, K, L, 100 P, Q, R, S, T or V; I at the 6th position is replaced with A, F, L, M, Q, R, S, T or V; Y at the 7th position is replaced with F, H, R or V; or G at the 8th position is replaced with D, F, H, M, N, R, S, T or V; ( i i ) G at the 1st position of the light chain CDR2 is replaced with D, F, H, K, P, Q, S, V or Y; T at the 3rd position is replaced with Q; or N at the 4th position is replaced with ; ( i i i ) Q at the 1st position of light chain CDR3 is E, G, I, M or N; 凡 at the 2nd position is A, D, E, H, L, Q, S or T; V at the 3rd position is I, L, M, N, Q, S or T; L at the 4th position is F, H, I, M, R, S, V, W or Y; S at the 5th position is C, D, E, F, G, H, K, L, N, Q, R, T, V or Y; S at the 6th position is D, E, F, G, H, I, L, M, N, P, Q, R, T, V or Y; P at the 7th position is A, D, E, G, N, Q, S or V; Y at the 8th position is replaced with E, F, L, M or Q; or T at the 9th position is replaced with D, F, G, I, L, N, S, V, W or Y; (iv) 日 at the 1st position of heavy chain CDR1 is replaced with G or Q; Y at the 2nd position is replaced with Q; or I at the 4th position is replaced with A or I; (v) The 3rd position of the heavy chain CDR2 is D, E, W, or Y; the 5th position is D, H, or Y; the 6th position is F, P, W, or Y; the G at the 7th position is A, F, L, N, or T; the 8th position is F, P, S, T, or Y; the T at the 9th position is A, D, E, F, G, H, L, P, S, or V; the H at the 10th position is A, D, F, M, R, S, T, V, W, or Y; the F at the 11th position is G, H, I, L, M, N, P, Q, V, or Y; the S at the 12th position is A, D, G, H, I, L, P, T, or V; the A at the 13th position is D, E, F, G, H, I, K, L, M, P, R, S, T, V, or Y; R in the 14th position is replaced by A, E, G, H, L, N, P, Q, S, W or Y; F in the 15th position is replaced by D, E, G, L, M, P, R, S, V or W; V in the 16th position is replaced by A, E, F, G, H, L, R, S, T, V or Y; or G in the 17th position is replaced by E, F, H, L, M, N, P, Q, R, S, T, V or W; or 101 (vi) A pharmaceutical composition wherein G at the 1st position of the heavy chain CDR3 is replaced with E, F, H, N, Q, V, or W; 日 at the 2nd position is replaced with E; 오 at the 3rd position is replaced with L, Q, T, or V; 으 at the 4th position is replaced with W; F at the 5th position is replaced with L or Y; L at the 6th position is replaced with Q, S, or Y; or Y at the 7th position is replaced with C, L, M, N, or Y, wherein the light chain CDR1 comprises 0 to 5 substitutions, the light chain CDR2 comprises 0 to 1 substitution, the light chain CDR3 comprises 0 to 7 substitutions, the heavy chain CDR1 comprises 0 to 1 substitution, the heavy chain CDR2 comprises 0 to 11 substitutions, and the heavy chain CDR3 comprises 0 to 6 substitutions. 【Claim 31】 A pharmaceutical composition according to claim 27, wherein the antibody further comprises an antibody or an antigen-binding fragment thereof that specifically binds to epidermal growth factor receptor (EGFR). 【Claim 3: A pharmaceutical composition according to claim 27, wherein the antibody or antigen-binding fragment thereof comprises a light chain variable region as set forth in SEQ ID NO: 11 and any one of SEQ ID NOs: 288 to 293; and a heavy chain variable region as set forth in any one of SEQ ID NOs: 298 to 312. 【Claim 33】 In claim 27, the antibody or antigen-binding fragment thereof has SEQ ID NO: 11. A pharmaceutical composition comprising a light chain variable region comprising an amino acid sequence described in SEQ ID NO: 102 and a heavy chain variable region comprising an amino acid sequence described in SEQ ID NO: 308. 【Claim 34】 A pharmaceutical composition according to claim 27, wherein the cancer is a cancer that overexpresses c-Met, amplifies c-Met, or activates c-Met. 【Claim 35】 A method for preventing or treating cancer, comprising administering to a subject in need of prevention or treatment of cancer (A) an antibody or antigen-binding fragment thereof that specifically binds to hepatocyte growth factor receptor (c-Met) that binds to at least one epitope region described in an amino acid sequence selected from the group consisting of SEQ ID NO: 313, SEQ ID NO: 314, SEQ ID NO: 315, and SEQ ID NO: 316, wherein the antibody or antigen-binding fragment thereof is administered in combination with (B) at least one of the following (1) Drug 1 and (2) Drug 2: (1) Drug 1: SN-38 represented by the following chemical formula 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof [Chemical Formula 1] 103

(2) Drug 2: Irinotecan represented by the following chemical formula 2, an optical isomer thereof, or a pharmaceutically acceptable salt thereof [Chemical Formula 2]

[Claim 36] (A) An antibody or antigen-binding fragment thereof that specifically binds to hepatocyte growth factor receptor (c-Met) that binds to at least one epitope region described in an amino acid sequence selected from the group consisting of SEQ ID NO: 313, SEQ ID NO: 314, SEQ ID NO: 315 and SEQ ID NO: 316, for use in preventing or treating cancer, wherein the antibody or antigen-binding fragment thereof is used in combination with at least one of (1) Drug 1 and (2) Drug 2 below. 104 administered, intended for: (1) Drug 1: SN-38 represented by the following chemical formula 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof [Chemical Formula 1]

(2) Drug 2: Irinotecan represented by the following chemical formula 2, an optical isomer thereof, or a pharmaceutically acceptable salt thereof [Chemical Formula 2]

【Claim 37】 (A) in the manufacture of a drug for use in preventing or treating cancer 105 Use of an antibody or antigen-binding fragment thereof that specifically binds to hepatocyte growth factor receptor (c-Met) that binds to at least one epitope region as described by an amino acid sequence selected from the group consisting of SEQ ID NO: 313, SEQ ID NO: 314, SEQ ID NO: 315 and SEQ ID NO: 316, wherein the antibody or antibody fragment is administered in combination with at least one of (1) Drug 1 and (2) Drug 2 below: (1) Drug 1: SN-38 represented by the following chemical formula 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof [Chemical Formula 1]

(2) Drug 2: Irinotecan represented by the following chemical formula 2, an optical isomer thereof, or a pharmaceutically acceptable salt thereof [Chemical Formula 2] 106