EP-4740966-A1 - THERAPEUTIC USE OF PROTEIN HETERODIMER

EP4740966A1EP 4740966 A1EP4740966 A1EP 4740966A1EP-4740966-A1

Abstract

Use of a protein heterodimer in the preparation of a drug, which drug is used for preventing and/or treating malignant effusion. The protein heterodimer contains a first member and a second member different from the first member, wherein the first member contains a light chain and a heavy chain, the heavy chain contains a first Fc region, and the light chain is complexed with the heavy chain to form a targeting moiety that exhibits binding specificity for a tumor antigen; and the second member contains a polypeptide containing an immunomodulator fused to a second Fc region. The first member and the second member are associated by means of the complexing of the first Fc region with the second Fc region so as to form the heterodimer. In addition, the first Fc region contains a first modification and/or the second Fc region contains a second modification, and these modifications can more effectively promote heterodimerization between the first member and the second member.

Inventors

ZHANG, XIN
PAN, Xiaolong
FU, Shilong
ZHOU, Liyao

Assignees

Shihuida Pharmaceutical Group (Jilin) Co., Ltd.

Dates

Publication Date: 20260513
Application Date: 20240702

Claims (20)

Use of a proteinaceous heterodimer in the manufacture of a medicament for preventing and/or treating a malignant effusion, wherein the proteinaceous heterodimer comprises a first member and a second member different from the first member, wherein: i. the first member comprises a light chain and a heavy chain comprising a first Fc region, and the light chain is complexed with the heavy chain to form a targeting moiety exhibiting binding specificity to a tumor antigen; ii. the second member comprises a polypeptide comprising an immunoregulator fused to a second Fc region; iii. the first member associates with the second member to form the heterodimer through complexation of the first Fc region with the second Fc region; and iv. the first Fc region comprises a first modification and/or the second Fc region comprises a second modification, wherein the first modification and/or the second modification more effectively promotes heterodimerization between the first member and the second member than a knob-and-hole modification comprising a knob modification and a hole modification.
The use according to claim 1, wherein the polypeptide comprised in the second member is a fusion protein, and a C-terminus of the immunoregulator is fused to an N-terminus of the second Fc region to form the fusion protein, optionally via a linker.
The use according to any one of the preceding claims, wherein the tumor antigen is selected from the group consisting of EGFR, an EGFR mutant, HER2/neu, GPC3, FAP, Muc1, MUC5AC, and Mesothelin.
The use according to any one of the preceding claims, wherein the immunoregulator is a cytokine selected from the group consisting of an interferon, an interleukin, a chemokine, a lymphokine, and a tumor necrosis factor.
The use according to any one of the preceding claims, wherein the immunoregulator is an interferon selected from the group consisting of interferon alpha, interferon lambda, and interferon beta.
The use according to any one of the preceding claims, wherein the immunoregulator is an interleukin, and the interleukin comprises interleukin 10, interleukin 2, and/or super interleukin 2.
The use according to any one of the preceding claims, wherein the first Fc region and the second Fc region are from an Fc region of an immunoglobulin, and the immunoglobulin is a human IgG1.
The use according to any one of the preceding claims, wherein the polypeptide comprised in the second member comprises two or more immunoregulators, the two or more immunoregulators are fused in frame to each other and to the second Fc region, and wherein the two or more immunoregulators are located N-terminal to the second Fc region.
The use according to any one of the preceding claims, wherein the first modification comprises an amino acid substitution at position T366, and an amino acid substitution at one or more positions selected from the group consisting of: Y349, F405, K409, D399, K360, Q347, K392, and S354, wherein the position of the amino acid is determined according to the EU index of the KABAT number.
The use according to claim 9, wherein the amino acid substitution comprised by the first modification is selected from the group consisting of Y349C, Y349D, D399S, F405K, K360E, K409A, K409E, Q347E, Q347R, S354D, K392D, and T366W.
The use according to any one of claims 9-10, wherein the first modification comprises an amino acid substitution at a group of positions selected from any of the following groups: 1) Y349 and T366; 2) Y349, T366, and F405; 3) Y349, T366, and K409; 4) Y349, T366, F405, K360, and Q347; 5) Y349, T366, F405, and Q347; 6) Y349, T366, K409, K360, and Q347; 7) Y349, T366, K409, and Q347; 8) T366, K409, and K392; 9) T366 and K409; 10) T366, K409, Y349, and S354; 11) T366 and F405; 12) T366, F405, and D399; and 13) T366, F405, Y349, and S354.
The use according to any one of claims 9-11, wherein the first modification comprises a group of amino acid substitutions selected from any of the following groups: 1) Y349C and T366W; 2) Y349C, T366W, and F405K; 3) Y349C, T366W, and K409E; 4) Y349C, T366W, and K409A; 5) Y349C, T366W, F405K, K360E, and Q347E; 6) Y349C, T366W, F405K, and Q347R; 7) Y349C, T366W, K409A, K360E, and Q347E; 8) Y349C, T366W, K409A, and Q347R; 9) T366W, K409A, and K392D; 10) T366W and K409A; 11) T366W, K409A, and Y349D; 12) T366W, K409A, Y349D, and S354D; 13) T366W and F405K; 14) T366W, F405K, and D399S; 15) T366W, F405K, and Y349D; and 16) T366W, F405K, Y349D, and S354D.
The use according to any one of claims 9-12, wherein the second modification comprises amino acid substitutions at positions T366, L368, and Y407, as well as an amino acid substitution at one or more positions selected from the group consisting of D356, D399, E357, F405, K360, K392, K409, and Q347, wherein the position of the amino acid is determined according to the EU index of the KABAT number.
The use according to claim 13, wherein the amino acid substitution comprised by the second modification is selected from the group consisting of D356C, D399S, E357A, F405K, K360E, K392D, K409A, L368A, L368G, Q347E, Q347R, T366S, Y407A, and Y407V.
The use according to any one of claims 13-14, wherein the second modification comprises an amino acid substitution at a group of positions selected from any of the following groups: 1) D356, T366, L368, Y407, and F405; 2) D356, T366, L368, and Y407; 3) D356, T366, L368, Y407, and Q347; 4) D356, T366, L368, Y407, K360, and Q347; 5) D356, T366, L368, Y407, F405, and Q347; 6) D356, T366, L368, Y407, F405, K360, and Q347; 7) T366, L368, Y407, D399, and F405; 8) T366, L368, Y407, and F405; 9) T366, L368, Y407, F405, and E357; 10) T366, L368, Y407, and K409; 11) T366, L368, Y407, K409, and K392; and 12) T366, L368, Y407, K409, and E357.
The use according to any one of claims 13-15, wherein the second modification comprises a group of amino acid substitutions selected from any of the following groups: 1) D356C, T366S, L368A, Y407V, and F405K; 2) D356C, T366S, L368A, and Y407V; 3) D356C, T366S, L368A, Y407V, and Q347R; 4) D356C, T366S, L368A, Y407V, K360E, and Q347E; 5) D356C, T366S, L368A, Y407V, F405K, and Q347R; 6) D356C, T366S, L368A, Y407V, F405K, K360E, and Q347E; 7) T366S, L368A, Y407V, D399S, and F405K; 8) T366S, L368G, Y407A, and F405K; 9) T366S, L368A, Y407V, F405K, and E357A; 10) T366S, L368A, Y407V, and K409A; 11) T366S, L368A, Y407V, K409A, and K392D; 12) T366S, L368G, Y407A, and K409A; 13) T366S, L368A, Y407V, K409A, and E357A.
The use according to any one of the preceding claims, wherein the first Fc region comprises the first modification, the second Fc region comprises the second modification, and the first modification and the second modification comprise an amino acid substitution at a group of positions selected from any of the following groups: 1) the first modification: Y349 and T366; and the second modification: D356, T366, L368, Y407, and F405; 2) the first modification: Y349, T366, and F405; and the second modification: D356, T366, L368, and Y407; 3) the first modification: Y349, T366, and K409; and the second modification: D356, T366, L368, Y407, and F405; 4) the first modification: Y349, T366, F405, K360, and Q347; and the second modification: D356, T366, L368, Y407, and Q347; 5) the first modification: Y349, T366, F405, and Q347; and the second modification: D356, T366, L368, Y407, K360, and Q347; 6) the first modification: Y349, T366, K409, K360, and Q347; and the second modification: D356, T366, L368, Y407, F405, and Q347; 7) the first modification: Y349, T366, K409, and Q347; and the second modification: D356, T366, L368, Y407, F405, K360, and Q347; 8) the first modification: T366, K409, and K392; and the second modification: T366, L368, Y407, D399, and F405; 9) the first modification: T366 and K409; and the second modification: T366, L368, Y407, and F405; 10) the first modification: T366, K409, and Y349; and the second modification: T366, L368, Y407, F405, and E357; 11) the first modification: T366, K409, Y349, and S354; and the second modification: T366, L368, Y407, F405, and E357; 12) the first modification: T366 and F405; and the second modification: T366, L368, Y407, and K409; 13) the first modification: T366, F405, and D399; and the second modification: T366, L368, Y407, K409, and K392; 14) the first modification: T366, F405, and Y349; and the second modification: T366, L368, Y407, K409, and E357; 15) the first modification: T366, F405, Y349, and S354; and the second modification: T366, L368, Y407, K409, and E357; wherein the position of the amino acid is determined according to the EU index of the KABAT number.
The use according to any one of the preceding claims, wherein the first Fc region comprises the first modification, and the second Fc region comprises the second modification, wherein the first modification and the second modification comprise a group of amino acid substitutions selected from any of the following groups: 1) the first modification: Y349C and T366W; and the second modification: D356C, T366S, L368A, Y407V, and F405K; 2) the first modification: Y349C, T366W, and F405K; and the second modification: D356C, T366S, L368A, and Y407V; 3) the first modification: Y349C, T366W, and K409E; and the second modification: D356C, T366S, L368A, Y407V, and F405K; 4) the first modification: Y349C, T366W, and K409A; and the second modification: D356C, T366S, L368A, Y407V, and F405K; 5) the first modification: Y349C, T366W, F405K, K360E, and Q347E; and the second modification: D356C, T366S, L368A, Y407V, and Q347R; 6) the first modification: Y349C, T366W, F405K, and Q347R; and the second modification: D356C, T366S, L368A, Y407V, K360E, and Q347E; 7) the first modification: Y349C, T366W, K409A, K360E, and Q347E; and the second modification: D356C, T366S, L368A, Y407V, F405K, and Q347R; 8) the first modification: Y349C, T366W, K409A, and Q347R; and the second modification: D356C, T366S, L368A, Y407V, F405K, K360E, and Q347E; 9) the first modification: T366W, K409A, and K392D; and the second modification: T366S, L368A, Y407V, D399S, and F405K; 10) the first modification: T366W and K409A; and the second modification: T366S, L368G, Y407A, and F405K; 11) the first modification: T366W, K409A, and Y349D; and the second modification: T366S, L368A, Y407V, F405K, and E357A; 12) the first modification: T366W, K409A, Y349D, and S354D; and the second modification: T366S, L368A, Y407V, F405K, and E357A; 13) the first modification: T366W and F405K; and the second modification: T366S, L368A, Y407V, and K409A; 14) the first modification: T366W, F405K, and D399S; and the second modification: T366S, L368A, Y407V, K409A, and K392D; 15) the first modification: T366W and F405K; and the second modification: T366S, L368G, Y407A, and K409A; 16) the first modification: T366W, F405K, and Y349D; and the second modification: T366S, L368A, Y407V, K409A, and E357A; 17) the first modification: T366W, F405K, Y349D, and S354D; and the second modification: T366S, L368A, Y407V, K409A, and E357A; wherein the position of the amino acid is determined according to the EU index of the KABAT number.
The use according to any one of the preceding claims, wherein the first Fc region comprises the first modification, the second Fc region comprises the second modification, the first modification comprises the amino acid substitutions T366W and K409A, and the second modification comprises the amino acid substitutions T366S, L368G, Y407A, and F405K, wherein the position of the amino acid is determined according to the EU index of the KABAT number.
The use according to any one of the preceding claims, wherein the targeting moiety specifically binds to EGFR, the light chain of the first member comprises CDR1-3, the amino acid sequence of the CDR1 is as set forth in SEQ ID NO: 101, the amino acid sequence of the CDR2 is as set forth in SEQ ID NO: 102, and the amino acid sequence of the CDR3 is as set forth in SEQ ID NO: 103; the heavy chain of the first member comprises CDR1-3, the amino acid sequence of the CDR1 is as set forth in SEQ ID NO: 105, the amino acid sequence of the CDR2 is as set forth in SEQ ID NO: 106, and the amino acid sequence of the CDR3 is as set forth in SEQ ID NO: 107.

Description

TECHNICAL FIELD The present disclosure relates to the field of biomedicine, and in particular to therapeutic use of a proteinaceous heterodimer. BACKGROUND Malignant pleural effusion (MPE) is one of the most common complications of advanced metastatic cancer, which is caused by pleural metastasis and contains malignant cells. Tumor cells infiltrate the pleural space by direct infiltration, hematogenous metastasis, or lymphatic diffusion, and pleural effusion may be the result of tumor growth obstructing lymphatic drainage. Malignant ascites is a clinical manifestation of advanced stages of various malignant tumors, which generally occurs at the late stage of the disease. The occurrence of malignant ascites is often accompanied by distant metastasis of malignant tumors, at which point drug treatment has become difficult. Therefore, the formation of ascites indicates that there is generally a lack of effective treatment methods in clinical practice. The formation of ascites is particularly prominent in patients with advanced ovarian, pancreatic, and gastrointestinal malignant tumors. At present, the clinical treatment mainly includes: simple ascites extraction, intra-abdominal injection of chemotherapeutic drugs, hyperthermic intraperitoneal perfusion therapy, and the like. However, the means described above are often accompanied by significant toxic effects. Therefore, there is an urgent need to obtain an effective method for treating malignant pleural effusion and malignant ascites. SUMMARY The present disclosure provides use of a proteinaceous heterodimer, a mixture for producing the proteinaceous heterodimer, a nucleic acid encoding the proteinaceous heterodimer, and the like in the treatment of a malignant effusion, such as malignant pleural effusion and malignant ascites. The use described herein provides high utilization efficiency and low side effects for the proteinaceous heterodimer, and has very broad application prospects. In one aspect, the present disclosure provides use of a proteinaceous heterodimer in the manufacture of a medicament for preventing and/or treating a malignant effusion, wherein the proteinaceous heterodimer may comprise a first member and a second member different from the first member, the first member may comprise a light chain and a heavy chain comprising a first Fc region, the light chain may be complexed with the heavy chain to form a targeting moiety exhibiting binding specificity to a tumor antigen; the second member may comprise a polypeptide comprising an immunoregulator fused to a second Fc region; the first member may associate with the second member to form the heterodimer through complexation of the first Fc region with the second Fc region; and the first Fc region may comprise a first modification and/or the second Fc region may comprise a second modification, wherein the first modification and/or the second modification may more effectively promote heterodimerization between the first member and the second member than a knob-and-hole modification comprising a knob modification and a hole modification. For the knob-and-hole modification comprising a knob modification and a hole modification, the first Fc region may comprise the knob modification, and the second Fc region may comprise the hole modification. Alternatively, the first Fc region may comprise the hole modification, and the second Fc region may comprise the knob modification. In some embodiments, the first modification is different from the knob modification or the hole modification, and/or the second modification is different from the knob modification or the hole modification. In some embodiments, when expressed in a mammalian cell, a yield of the proteinaceous heterodimer is at least 10% higher than that of a reference protein, and the reference protein differs from the proteinaceous heterodimer in that the reference protein: i) comprises the knob modification in the first Fc region, ii) comprises the hole modification in the second Fc region, and iii) does not comprise the first modification and the second modification of the proteinaceous heterodimer of the present disclosure simultaneously. The mammalian cell may be selected from the group consisting of a HEK293 cell, a CHO cell, a COS-1 cell, and an NS0 cell. In some embodiments, the first Fc region comprises the first modification, the second Fc region comprises the second modification, and neither the first modification nor the second modification is the same as the knob modification or the hole modification. In some embodiments, the polypeptide comprised in the second member is a fusion protein, and a C-terminus of the immunoregulator is directly or indirectly fused to an N-terminus of the second Fc region to form the fusion protein. In some embodiments, the tumor antigen is selected from the group consisting of EGFR, an EGFR mutant, HER2/neu, GPC3, FAP, Muc1, MUC5AC, and Mesothelin. In some embodiments, the light chain of the targeting moiety